Newcastle disease : a review of some of the literature published between 1926 and 1964
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NEWCASTLE DISEASE A REVIEW
1926-1964
J
. E .
Lancaster
ALBERT R. MANN
LIBRARY New York State Colleges Agriculture
OF
and Home Economics
AT
Cornell University
DISEASE
NEWCASTLE
A REVIEW OF SOME OF THE LITERATURE
PUBLISHED
BETWEEN
John
1926 AND
1964
E. Lancaster
1966 Health of Animals
Branch
CANADA DEPARTMENT OF AGRICULTURE
Monograph
No. 3
373333 © Crown Copyrights reserved Available by mail from the Queen's Printer, Ottawa, and at the following Canadian Government bookshops: OTTAWA
Daly Building, Corner Mackenzie and Rideau TORONTO
Mackenzie Building, 36 Adelaide St. East MONTREAL
AEterna-Vie Building 1182 St. Catherine St. West WINNIPEG
Mall Center Bldg.,
499 Portage Avenue
VANCOUVER 657 Granville Street
or through your bookseller
A
deposit copy of this publication is also available for reference in public libraries across Canada
Price $2.00
Catalogue No. A63-1254
Price subject to change without notice
Roger Duhamel, f.r.s.c. Queen's Printer and Controller of Stationery Ottawa, Canada 1966
ii
FOREWORD Newcastle research
disease has probably
workers
received
than any other respiratory
more
attention
from poultrymen
disease of poultry.
It
is worldwide,
and and
fowl that are reared in thousands by modern intensive methods as well as individual fowl that seek their own food in rural areas. Control of the disease is essential if the poultry industry of many countries is to flourish. In this book Dr. Lancaster reviews progress that has been made to date towards accomplishing this goal. It is not a book that is likely to be of immediate interest to most poultrymen, though it should benefit them indirectly. It is a research tool that should be valuable to anyone studying Newcastle disease or its virus. affects
K. F. WELLS Veterinary Director General Ottawa, 1965
iii
PREFACE This review is based on articles
I
have written
over the past several
years
for
publication in a number of different journals. In bringing all these articles together into one volume I have expanded on them and also added a number of tables and diagrams.
The material covers many aspects of research on Newcastle disease and its virus, but
I
have made no attempt to discuss every report that has been published on these subjects. I have read most of the reports referred to in the original, and where this has not been possible
I
have used abstracts published by the Commonwealth
The Veterinary
Agricultural Bureaux in
Bulletin.
Mention must be made here of three sources of information on Newcastle disease of the main work on this book. In 1963, an international symposium on "Newcastle Disease Virus — An Evolving Pathogen" was held in Madison, Wisconsin, U.S.A., and the proceedings, edited by Dr. R. P. Hanson of the Study Center for Newcastle Disease at the University of Wisconsin, have now been published. Dr. Hanson has also compiled a subject bibliography on Newcastle disease for the period 1926 to 1962. This bibliography is in manuscript form. Dr. V. N. Syurin of the Soviet Union has published a book in Russian (Psevdochuma ptits [Newcastle Disease of Poultry] Moscow, 1963) in which he makes a detailed study of Russian and foreign literature on the characteristics of viruses, with special reference to Newcastle disease virus. Dr. Syurin's book includes a more detailed review of Russian work on Newcastle disease than is given here. I hope that the present volume, used in conjunction with these other works, will serve which have become available since completion
as a useful
reference
to the literature
on Newcastle
disease published
between
and 1964.
JOHN
E. LANCASTER
Ottawa, 1965
iv
1926
ACKNOWLEDGMENTS The publication of this review would not have been possible without the kind help of many individuals and organizations. Grateful acknowledgment is made to: The Commonwealth Agricultural Bureaux for permission to reproduce the con tents of three reviews by the author published in The Veterinary Bulletin (Vol. 33, pp. 221, 279 and 347, and Vol. 34, p. 57); Dr. R. A. Bankowski of the University of California at Davis, Dr. G. L. Bannister of the Animal Diseases Research Institute, Canada Department of Agriculture, Dr. A. P. Waterson of St. Thomas's Hospital, London, and the Controller of Her Majesty's Stationery Office, London, for providing photographs; Dr. A. S. Greig for modifying one of the drawings; and For permission to reproduce drawings and tabular material, the authors whose names are given in captions; the publishers of the following Journals: Acta Veterinaria Hungarica (Figure 19), American Journal of Hygiene (Figure 6), American Journal of Veterinary Research (Figure 15 and Table 7), Animal Health Yearbook (Figure 5), Annals of the New York Academy of Sciences (Table 20), Avian Diseases (Figure 7), Bacteriological Reviews (Figure 1), British Veterinary Journal (Figure 23), Bulletin de l'Office International des Epizooties (Table 2), Bulletin of the Interafrican Bureau for Animal Health (Figure 3), Canadian Journal of Comparative Medicine and Veterinary Science (Table 13), Cornell Veterinarian (Figure 13), Journal of Comparative Pathology and Therapeutics (Figures 11 and 18 and Tables 11 and 23), Journal of Experimental Medicine (Figure 14), Journal of Immunology (Figures 21 and 22), The Veterinary Bulletin (Tables 14, 15, 16 and 18), The Veterinarian (Table 10), The Veterinary Record (Figure 17 and Table 12), World's Poultry Science Journal (Table 5); and the American Veterinary Medical Association (Tables 3 and 22), the Controller of Her Majesty's Stationery Office, London (Table 1 ) , the Eighth World's Poultry Congress (Table 4), the Italian Society of Veterinary Science (Table 21), the National Academy of Sciences, Washington, D.C. (Table 8), and the University of Wisconsin Press, Madison, Wisconsin (Figure 20) . The author's sincere thanks also go to personnel of the Canada Department of to the staff of the Library for obtaining articles and reports for review Agriculture: purposes; to the Art Section of the Information Division and the bio-Graphic Unit of the Research Branch for preparing text figures; and, especially, to the Editorial Unit of the Information Division for suggesting improvements to the original manuscript and for attending to the many details of publication.
v
CONTENTS Page
Page Summaries in English and French Definition of Newcastle Disease Nomenclature Properties of the virus
1
....
13 13 13
Asymptomatic infections Transport of live poultry Poultry markets Laying trials
Geographic Distribution Early reports of diseases Newcastle
resembling
17
disease
First outbreaks of Newcastle disease in England Spread through Southeast Asia India Philippines Malaya Haiti and Madagascar United States Canada
17
18 ..
18
Europe
18 18 19
22 22
Austria World distribution Modes of Spread Wild birds Infections
migratory
birds
44
43
II — DIAGNOSIS of the 45
Breed
46
25
Sex differences in susceptibility
27
Ducks and geese
29
Turkeys
29
differences in resistance
46 46 47 48 48
physiology
29
Young chicks Growing and adult chickens Excretion of virus from
30
respiratory system
31
Haemagglutination
31
Haemagglutination-inhibition
33
Other
31
Diagnosis
by Serological
(HI vii
46 46
The Disease in Chickens Velogenic form Mesogenic form Lentogenic form form Asymptomatic The Disease in Turkeys The Disease in Ducks and Geese The Disease in Game Birds
27 28
susceptibility
Age susceptibility Effect of season of year Climatic influences Routes of infection Effect of the virus on avian
27
28
Excretion of virus in faeces Reservoir of virus in aqueous humour
44
Genetic
Pigeons and doves
eggs
Water
23
Game birds
Chicken
42
22
Introduction of virus by Excretion of virus in faeces
41
Wind Aerosols
45
25 birds
41
causes
Incubation period
Introduction of virus by imported
40
Inanimate
36
Disease
in birds in zoological
gardens
38
General Characteristics
22 in 1962
Human infection Warm-blooded animals Cold-blooded animals
PART
19
Ireland
36
Poultry vaccines as a means of spread of Newcastle disease
19
Great Britain
34
Spread by human agency
brooders
18
19
Spread through
34
Poultry carcasses and offal
Chicken houses, crates and
19
Africa
33
34
movement
Illegal
PART I — SPREAD OF THE DISEASE
33 33
48 48 53
56 57 57 58 58
Methods
(HA
Test)
59 ..
61
Test) haemagglutinating
59
agents
63
Page
Page Haemolysis
63
Intradermal
inoculation
64
Fluorescent
antibody
64
Serum electrophoresis
64
Serum or virus neutralization 64
(SN Test) Complement-fixation
66
tests
Precipitation test by Virus Isolation Distribution of virus in the body Embryonating eggs Preparation of inoculum Route of inoculation Temperature of egg incubation Blind passages Embryonic mortality
92
Age at time of vaccination
93
68
Virus titre of the vaccine Viral interference
93
69 69
Differences birds
by Inoculation
73
Pigeons
73
Ducks Laboratory mammals
74
Exposure
of Newcastle
Identification
Bacterial
Bl
74
Effectiveness
Sterilization
100
LaSota strain
102 strains
102
Komarov (or Haifa) strain Mukteswar strain Strains originating
78
United
78
103
in the
States
104
strain
105
Other mesogenic strains
80
106
Extract of tobacco mosaic virus
106
Tissue culture vaccines
106
Inactivated
vaccines
106
Inactivation by beta-
84
propiolactone Inactivation
with Newcastle
106
by formalin
Inactivation by crystal violet ..
102
Hertfordshire (or Herts)
on Newcastle
Disease Virus
102
strains
Mesogenic
80
Control with Hyperimmune Serum
98
F
Other lentogenic
of Atmospheres
Contaminated
97
strain
Strain
78
in various countries
97 strains
Lentogenic
III — CONTROL MEASURES
Action of Chemicals Disease Virus
95
compared
78
Control by Slaughter
95
Live vaccines
75
Nutritional deficiencies Toxic drugs and plants Avian respiratory diseases
95
Live and inactivated vaccines
75
diseases
— Administration
and Effectiveness
74
Differential Diagnosis Viral diseases
PART
immunity
Disease
Virus
95
Types of Vaccines
74 ....
to another
Other factors that affect
73
Chickens
on
disease
70 70
by Challenge
95
susceptibility
72
93
between individual
Effect of vaccination
70 eggs
92
67
Mixed virus infections
Diagnosis
of immunity
67
Tissue cultures Diagnosis
89
Passive immunity
Distribution of virus in De-embryonated
89
development
69
embryos
89
67
69
Lesions in embryos
88
Factors that influence
67 .
88
immunity
66
Diagnosis
Control with Hyperimmune Serum Combined with Virus Control with Antibiotics and Other Medicinal Agents Control by Vaccination Antigenic plurality Procedures for evaluating
110 ....
113
87
Inactivation
by other chemicals
113
87
Inactivation
by heat
113
viii
Page
Page
Inactivation by ultraviolet irradiation Inactivation by ultrasonic
Preparation
embryonating
113
treatment
of parental immunity of vaccines .... 125 127 Storage of virus material Use of extra-embryonic fluids with or without suspensions of embryonic tissues 127 Preparation of fresh and 129 lyophilized vaccine Preparation of inactivated vaccine 129 Preparation of Newcastle disease vaccines from virus propagated on preparation
114 disease
vaccine
114
Combined vaccines
114
PART IV — VIRUS PROPAGATION AND VACCINE PRODUCTION Methods
116
Propagation
in Eggs
116
Propagation
in avian hosts
117
Propagation
in mammalian
Propagation
in tissue cultures
Propagation
in yeast cells
hosts ..
in tissue culture
130
Storage and Transportation of
118
Vaccines
119
130
Testing and Standardization
124
Preparation of Vaccines
125
Influence
114
methods compared
Virus Propagation
125
eggs
Egg transmitted diseases
Vaccines inactivated by different "Incomplete" Newcastle
of vaccines from
of
Vaccines
133
REFERENCES
125
135
ILLUSTRATIONS Figure 1. Scheme
Newcastle 2.
disease virus
Newcastle
disease virus
south of the Sahara
20
1940-1955
Incidence of Newcastle
The velogenic form of Newcastle disease — distribution of lesions
52
12.
The mesogenic form of
the results of
14. Tissue distribution
of Newcastle
24
vaccination 32
vaccine
The velogenic form of Newcastle disease — symptoms of paralysis 9. The velogenic form of Newcastle
—
68
Selective infection by Newcastle
16.
Newcastle
17.
Number of outbreaks of Newcastle disease in Great Britain and Lancashire,
disease virus
49
lesions in
proventriculus
1 0-week-old
15.
47
8.
disease
of
chickens
Mortality due to vaccination with a wing-web
65
disease virus after intramuscular
Suggested scheme for the pathogenesis of Newcastle
7.
54
symptoms 21
disease virus
— nervous
A comparison of HI and SN tests
13.
disease
in Europe in 1962 6.
51
1 1.
Newcastle disease
Infection density of Newcastle disease and fowl plague in Europe,
5.
The velogenic form of Newcastle disease — lesions in intestine ....
15
disease in Africa
Page
10. 14
Electron micrograph of
3. Newcastle 4.
Figure
Page of the structure of
50
1946-1961
ix
disease in the pigeon .
71 73
82
Figure 18.
Immune response following with a BPL vaccine..
vaccination 19.
Figure
Page
non-immune
disease virus 21. Amount of
NDV (Bl
in
allantoic fluid, blood and 121
embryo tissues following
123
non-immune
of Newcastle
20. Cytopathogenicity
127
embryos
NDV (CGI 79)
22. Amount of
108
Newcastle disease virus in avian tissue culture
Page
of
allantoic sac inoculation
128
embryos
23. Viability of Komarov strain
strain) in
allantoic fluid following
Newcastle
allantoic sac inoculation of
vaccine
disease lyophilized 132
TABLES Page
Page 1. Outbreaks
of fowl pest in Great
Britain, 1954-1961 2.
Incidence and control of
3.
Wild birds susceptible
Newcastle
disease, 1962
experimentally
22
Duration of immunity following vaccination of chicks up to five weeks of age with Bl vaccine
23
virus
14.
15.
to Newcastle
disease virus and methods of evaluation 4.
26
1947
16.
6.
7.
34
Newcastle
Newcastle 8.
35
production
36
infectivity of Newcastle
diseases
—
some characteristics 1 1. Effect
Newcastle 12.
1ll
19.
Tissues used for in vitro
13.
Effect
of intranasal
disease vaccination
through 76
Strain
F
126
eggs
of Newcastle
21. Survival
disease
virus at different
temperatures 22.
85
of test requirements for live and modified live virus
23. Summation
Newcastle
131
Summary
Newcastle
86
disease vaccines
of
133
32 potency tests
on one batch of inactivated
on chicks
from immune parents
1 19
20. Diseases of birds transmitted
Action of formalin on Newcastle disease virus
of Newcastle disease
propagation
on
disease virus
and 109
79
of disinfectants
in unvaccinated
virus
of classical fowl
Avian respiratory
disease on egg
Duration of immunity following initial vaccination of chickens with formalin-inactivated virus ..
plague and typical Newcastle 10.
107
1 8.
disease
disease compared
vaccine
BPL-vaccinated flocks
43
Characteristics
101
Comparison of effect of Newcastle
Effect of temperature on virus
9.
17.
42
disease virus
virus
Duration of immunity following initial vaccination of chickens with beta-propiolactone inactivated
disease virus isolated
from poultry carcasses imported into England in 1949 Reports of duration of viability of Newcastle disease virus in a variety of tissues Duration of viability of
Duration of immunity following vaccination of chicks up to five weeks of age with Strain F vaccine
Probable origin of the first 542 outbreaks of Newcastle disease in England and Wales during
5.
99
Newcastle
91
x
disease vaccine
134
SUMMARY
SOMMAIRE Maladie
de Newcastle
Pseudo-peste aviaire
Spread of the Disease
Propagation de la Maladie
Acute diseases of poultry that resemble Newcastle disease have been recorded since early in the eighteenth century, but it is generally accepted that Newcastle disease, as now recognized, first appeared in epidemic form on the island of Java in in 1926. From this origin,
Indonesia
the
Depuis
le début du dix-huitième
on enregistre
des
volaille ressemblant
siècle,
aiguës de la
maladies
de New
à la maladie
castle. Cependant ce n'est qu'en 1926 sur l'île de Java, en Indonésie, que s'est mani festée la première épidémie de maladie de Newcastle
telle qu'on
la connaît
aujour
disease spread with a rapidity not previous
d'hui. De ce foyer, la maladie s'est répan
among diseases of poultry. Much of the early global spread was associated with coastal ports or towns, and the disease takes its name from the coastal town of Newcastle-on-Tyne in England. One of the exceptions to this general pat tern was the appearance of the disease in 1927 at the small town of Ranikhet in the Kumaon Hills of India. Another event of considerable importance in the history of the disease was the appearance in Cali fornia, around 1935, of a mild respiratory
due avec une rapidité inconnue auparavant
l'histoire
de la maladie:
disease
piratoire
peu sérieuse, appelée pneumoen-
ly recorded
itis".
termed
Several
"avian years
pneumoencephal-
elapsed
disease was recognized
as a
before
this
form of New
dans les annales des maladies aviaires.
La maladie tire son nom de la ville por en Angle de Newcastle-on-Tyne, terre. Elle a été propagée surtout en raison des volailles dans les de la manutention ports et les villes côtières, mais il y a eu, tuaire
aussi,
isolées, manifestations par dans la petite ville de Raniket,
des
exemple
dans les collines de Kumaon, aux Indes en 1927. Autre événement
important
dans
une affection res
céphalite aviaire se produisit en Californie, vers 1935; il s'est écoulé plusieurs années
castle disease.
avant qu'elle
Many of the early outbreaks of New castle disease were associated with deaths of free-flying birds in the vicinity. How ever, it was not until about 1 950 that the
forme de maladie de Newcastle. Au début, les manifestations de la maladie
virus
dans les envirous. Toutefois, ce n'est qu'en
was recovered
from free-flying (Columba livid), garis),
with
birds
starlings
sparrows
(Passer
pheasants
any regularity
such
pigeons
as
(Sturnus vul domesticus), and
(Phasianidae) (Perdix sp.). There is little evidence of partridges
Newcastle
disease existing in an endemic form in wild bird populations. Neverthe less, wild birds have played a part in the dissemination
of the disease both from one
to another and within a country. Chickens are more susceptible to the
country clinical
disease
than
other
domestic
poultry. However, even in chickens manent carrier state
is uncommon.
a per
Tur-
de Newcastle par la mort
soit reconnue
ont souvent d'oiseaux
comme
été
volant
une
marquées en liberté
1950 qu'on a constaté la présence régu lière du virus chez des pigeons (Columba
livid),
étourneaux
(Sturnus
vulgaris),
faisans (Passer domesticus). et perdix (Perdix sp.). Il (Phasianidae) existe peu de traces de la maladie de New moineaux
castle
sous
forme
endémique
oiseaux sauvages; néanmoins,
chez
les
les oiseaux
ont joué un rôle dans la dissémination de la maladie tant d'un pays à l'autre qu'à l'intérieur
de pays déterminés.
Les poulets sont plus susceptibles à la maladie que les autres volailles domesti ques. Toutefois, il est rare de trouver
un 1
keys are susceptible outbreaks
in
subclinical. healthy
this
to the disease, but
species
Because
often
of this, apparently-
have contributed
turkeys
remain to the
spread of Newcastle disease between coun tries and also within a country. Spread of the disease between individual birds and between premises can occur in a variety
of ways. Movement
of domestic
poultry
has been considered
the most im
portant
single cause of spread. Although
the virus has been recovered from the con tents of eggs laid by an infected flock, at the present time egg transmission of the virus is seldom reported. However, this
form of transmission remains potentially very dangerous; particularly as modern in the poultry industry have
developments meant
that newly
eggs,
chickens
numbers
large
hatched
chicks
are frequently
of hatching and growing
distributed
from
one central place.
The virus has survived outside the host for variable, and often long, periods. As a result, it is generally considered that the survival of the virus in the environment an important
plays
part in perpetuating
the disease. The virus
and spreading
is
frequently recovered from the faeces of birds suffering from a natural outbreak or
from poultry vaccinated with certain live vaccines. Following excretion, the virus is transmitted
readily by air-borne
particles.
This air-borne spread by means of atmos pheric air currents occurs between in dividual
chickens
in a pen, and between
different pens and premises. Other
sources
of infection
related
to
porteur permanent de virus même chez les Les
poulets.
dindons
aussi
souvent non cliniques; dindons apparemment
à répandre la maladie de New castle d'un pays à l'autre, aussi bien qu'à
contribué
l'intérieur
d'un même pays.
La propagation de la maladie entre oiseaux et d'un poulailler à l'autre peut se faire de différentes façons; le transport volailles
a été
domestiques
fection
generally
and There
sometimes
virus can produce in humans. Human in
causes a conjunctivitis, an influenza-like illness.
is no evidence
to date of the exis-
le
venant d'un troupeau oùlamaladieexistait, on signale de nos jours très peu de cas de transmission fois,
cette
du virus par les oeufs. Toute forme
de
reste
transmission
un danger, surtout avec les prati
toujours
telles la dis
ques modernes de l'industrie
tribution fréquente, sur de vastes distances, de grandes quantités d'oeufs d'incubation, de poussins et de poulets en croissance.
Le virus a parfois vécu hors de son hôte pour des périodes de temps variables et souvent prolongées; on considère générale ment que sa survivance dans le milieu joue un rôle important la propagation
dans la perpétuation
et
On trouve
de la maladie.
souvent le virus dans les matières fécales d'oiseaux
infectés
volailles
ayant
naturellement vaccinées
été
ou avec
de des
vaccins vivants. Des excrétions, le virus se transmet facilement dans l'air ambiant;
La propagation
peut
alors
se
produire
entre les poulets d'un même parquet ou de différents
parquets
poulaillers
éloignés.
ainsi
maladie
disease
de
considéré
facteur le plus important. Quoique le virus ait été recouvré à l'intérieur d'oeufs pro
castle disease virus in poultry vaccines has also introduced the infection to previously disease-free areas. infections
des
c'est pourquoi
nent les marchés à volailles
Newcastle
pré
en bonne santé ont
those already mentioned include poultry markets, and the movement of poultry carcasses and poultry offal. Virulent New
natural
sont
disposés à la maladie mais les cas restent
D'autres
sources
le déplacement
qu'entre
des
d'infection compren
des volailles
de même que abattues et des
déchets d'abattage. Des virus actifs de la de Newcastle
contenus
dans des
vaccins à volailles ont aussi été des agents d'infection dans des régions auparavant exemptes de la maladie. Le virus de la maladie de Newcastle est transmissible aux humains. Généralement,
il cause une conjonctivite
et parfois
une
tence
of
the carrier
state
in man.
and domestic
mammals.
of laboratory
species
However, many
mammals
are sus
ceptible to the virus by inoculation. atory
studies
have
involved
Labor
bats,
cats,
hamsters, mice, monkeys and other species.
There is little evidence
as yet to indicate that mammals play a part in the spread of
the disease.
a review
of the literature publish 1926-64, it would
that Newcastle
disease virus
atteints de la maladie de Newcastle. Toute fois, par inoculation, on a réussi à repro duire la maladie chez plusieurs espèces de mammifères
utilisés aux laboratoires. Des expériences ont été faites sur des chauvessouris, des chats, des hamsters, des souris, des singes et autres espèces. Il n'a pas été prouvé que les mammifères un rôle dans la propagation de la maladie.
jouent
ed during the period appear
Quelques rapports seulement ont dé montré que des mammifères sauvages ou auraient été naturellement domestiques
encore
Diagnosis of the Disease From
ressemblant à l'influenza. Il n'a pas encore été prouvé que l'homme puisse être porteur de ce virus. indisposition
Only a few reports have indicated natural Newcastle disease infection in wild
has
Diagnostic de la Maladie
been able to change in virulence and adapt to its host. When first identified in 1926 the disease was extremely virulent and
D'après les documents publiés au cours de la période 1927-1964, il semblerait que
caused nearly 100 per cent mortality.
par modifié
form of the disease recognized
The
about ten
years later, called avian pneumoencephalitis, was less virulent. After another interval
of ten years, around outbreaks
1944, asymptomatic
were reported.
The disease is now considered
to have
four main forms: the velogenic (virulent), the mesogenic (less virulent), the lentogenic
le virus de la maladie de Newcastle à son hôte.
la première
sa virulence
ait fini
et par s'adapter
Lorsqu'elle a été identifiée pour fois en 1926, cette maladie
était extrêmement
violente
et causait
un
taux de mortalité de près de 100 p. 100. La forme de cette maladie reconnue à peu près dix ans plus tard sous le nom de pneumoencéphalite,
était moins virulente.
Après un autre intervalle
de dix ans, vers
1944, on signalait des infectations
asymp-
(mild) and the asymptomatic. The average incubation period is four to five
tomatiques.
days, although
se présente sous quatre formes principales:
considerable
variation
has
been reported.
On considère maintenant que la maladie vélogénique
(virulente),
Chickens
période d'incubation
est de quatre
Velogenic Form
jours avec des variations
This form of Newcastle disease may be but more typically it causes marked depression, increased rate of res piration, progressive weakness, diarrhoea,
Forme vélogénique
peracute,
and death within a few days. Mortality is usually over 90 per cent. Survivors gener ally exhibit neurotropic
involvement.
Lesions are mainly haemorrhagic The haemorrhages
inflammatory.
mésogénique
(moins virulente), lentogénique (bénigne) et asymptomatique. La moyenne de la à cinq
considérables.
Poulets Cette forme de la maladie de Newcastle peut être suraiguë, mais dans ses manifes tations
les
plus
typiques
dépression marquée,
elle cause une
une accentuation
rythme de la respiration,
du
une faiblesse pro
and
gressive, de la diarrhée et, après quelques
are
jours, la mort. Le taux de mortalité dépasse 3
found
usually petechial and are commonly
in the mucosa and submucosa of the proventriculus,
gizzard
and intestinal
tract.
Microscopically, the lesions are essentially in character.
necrotizing
Mesogenic
ease
is common
In
appears suddenly
production
in many parts of
a susceptible
ly. Respiratory
flock, the dis
and spreads rapid
distress, sharp drop in egg
and
diarrhoea
are common
Mortality varies considerably
symptoms.
tuellement observées dans la muqueuse et la sous-muqueuse du proventricule, du
is
also
affected:
marked
Sous
présentent
Cette dans
forme
se
plusieurs
trouve
communément du
parties
dans
Une
les
troupeaux
diarrhée et une forte réduction en sont les symptômes
prédis
difficile,
respiration
de
Le taux
communs.
varie mais il est généralement
de mortalité
moins élevé que dans le cas de la forme
the albumen.
souvent des symptômes de paralysie.
ual birds and between outbreaks. Similarly, there is marked
variation
in the organs
and tissues involved.
sudden
drop
in the egg
of laying flocks are common features of the lentogenic form. In adult production
fowls, mortality is negligible, and complete recovery usually occurs within one to eight weeks. There are usually neither haemorrhagic lesions. Microscopic examin
ation has shown the respiratory
lymphoid
infiltration
of
and nervous systems.
Les
This form is often diagnosed only by In the absence of clinical signs,
Serological
is based on serological
4
findings.
results have indicated that the
asymptomatic
lésions
et inflam
hémorragiques
considérablement
varient
matoires
d'un
Forme lentogénique Cette forme se signale souvent par une des troubles bénins de la
perte d'appétit, respiration la ponte. les poules
form of Newcastle
et une diminution soudaine de mortalité est négligeable chez
La
adultes qui prennent
d'une
disease
huit semaines
à
générale
pour
se
rétablir. Habituellement, a pas
de lésions
cérales. L'examen infiltrations
tèmes respiratoire
Form
chance.
diagnosis
notables tant dans la
coquille que dans l'albumen.
des
Asymptomatic
produit une diminution de la ponte
se
et des modifications
ment
nor visceral
présentent
entre les organes et les tissus touchés.
of appetite, mild respiratory
and
symptoms
Il
survivants
Les
individu à l'autre et d'une épidémie à l'autre. On constate des variantes marquées
Lentogenic Form Impairment
la
de la ponte
vélogénique.
The lesions of haemorrhage and inflam between individ
La
monde.
maladie apparaît subitement et se propage
changes occur both in the egg shell and in
mation vary considerably
le les
Forme mésogénique
form. Symptoms of paralysis are common
quality
intestinal.
lésions
essentielles de la nécrose.
caractéristiques
posés.
Not only does egg production decrease,
les
microscope,
rapidement
in survivors.
Les survivants
100.
nerveuses. Les lésions sont surtout hémorragiques et inflammatoires. Des pétéchies sont habi
but is generally lower than in the velogenic
egg
90 p.
gésier et du conduit
Form
This form the world.
habituellement
souffrent de complications
dans cette forme il n'y ou vis
hémorragiques microscopique
lymphoïdes
fait voir
dans les sys
et nerveux.
Forme asymptomatique Ce n'est souvent que par hasard que l'on à diagnostiquer cette forme. En
réussit
l'absence de signes cliniques,
le diagnostic
recherches
sérologiques:
est basé sur des
may
spread within a flock or may show
little or no evidence of spread.
geese
and turkeys are more re
sistant to Newcastle
It
résultats
obtenus
indiquent
la
que
asymptomatique de la maladie de Newcastle à tout le peut se propager troupeau ou se limiter à quelques sujets.
Ducks, Geese and Turkeys Ducks,
les
forme
disease than chickens.
is not usually possible to distinguish
the
Canards, Oies et Dindons Les canards, les oies et les dindons sont
four forms of the disease that occur in chickens. Apart from this general differ
plus résistants que les poulets; il est habi tuellement impossible de distinguer chez
ence, the disease in turkeys resembles that in chickens. Ducks and geese are more
s'attaquent aux poulets. Hormis cette dif
resistant than turkeys, usually undergoing symptomless infection and giving negative post mortem findings.
number
of serological
Of
of Newcastle
these, the haemagglutination-
can be inhibited
chief of which
is
by several Newcastle
disease antibody.
Two procedures for conducting test have been described:
canards
et
des din
Les
poulets.
oies y sont encore
les
plus
sans présenter
peut même donner
des
the
HI
alpha and beta.
On
utilise
l'épreuve (IH) est la
ing chickens or chicken embryos, has been used to identify both Newcastle disease virus and antibody.
Other serological methods include hae molysis inhibition, an intradermal test, the use of fluorescent antibody, complement fixation, and double diffusion plate tests.
nombre
de
d'inhibition-hémagglutination plus acceptée. Cette épreuve
est basée sur la propriété
virus d'agglutiner
que possède le
les globules
rouges du
sang des volailles et de plusieurs espèces de Plusieurs
mammifères. maladie
substances dont la
est un anticorps
principale
de
centrations
(SN), or virus involving either grow
certain
pour diagnostiquer la maladie de Newcastle. Parmi celles-ci
cedure, the antibody (serum) is diluted serially and mixed with a constant amount
of antigen (virus). The serum neutralization
un
méthodes sérologiques
l'épreuve
test,
des
Diagnostic par les' Méthodes Sérologiques
In the alpha procedure, the Newcastle dis ease virus is diluted serially and mixed with equal volumes of serum. In the beta pro
neutralization
à celle
résultats négatifs.
the red blood cells (haemagglutination) of birds and several species of mammals. The substances,
dons ressemble
virus et l'autopsie
inhibition (HI) test has been most widely adopted. This test is based on the property of Newcastle disease virus to agglutinate
agglutination
férence, la maladie de Newcastle
qui
de symptômes, ils sont souvent porteurs de
methods have
been used in the diagnosis disease.
de la maladie
résistants que les dindons;
Diagnosis by Serological Methods A
eux les quatre formes
Newcastle
du virus de la
peuvent
avoir
un
effet inhibitoire sur l'agglutination.
On connaît
IH.
deux procédés
pour
faire
Dans la méthode alpha, on
dilue le virus en une série de diverses con que l'on mélange avec une quantité égale de sérum. La méthode bêta, au contraire,
consiste à diluer l'anticorps
(sérum) en une série de diverses concentra tions pour les mélanger avec une quantité invariable
d'antigène (virus). de neutralisation
L'épreuve
(SN) ou de neutralisation des poulets
en croissance
du sérum
du virus faite sur ou sur des em
bryons de poussins est utilisée pour identi fier le virus aussi bien que l'anticorps. Autres méthodes sérologiques: hémolyse5
inhibition, épreuve intradermique,
Diagnosis by Virus Isolation Serological
methods
field evidence.
ciently accurate to confirm
new foci of infection
However, confirmed
suffi
are usually
should be
chickens,
fluorescents,
d'anticorps
pigeons,
laboratory
mammals and tissue culture methods have been found both efficient and economical.
Differential Diagnosis of symptoms
lesions associated with Newcastle
and
disease
has made differential
diagnosis
Among the infectious
diseases that need
difficult.
from Newcastle disease
to be differentiated
du
complément et épreuve de double diffusion sur plaque.
Diagnostic par l'Isolement du Virus Les
méthodes
habituellement posé
d'après
suffisent
sérologiques
à confirmer
le diagnostic
symptômes
les
Toutefois de nouveaux The great variety
utilisa
fixation
and identification
by recovery
of the virus. Embryonating chicken eggs are commonly used for virus isolations. Susceptible
tion
cliniques.
foyers d'infection
devraient
être vérifiés
identifiant
le virus. On utilise souvent des
en obtenant
et en
embryons de poussins pour isoler le virus. Des poulets, des pigeons, des mammifères de laboratoire
susceptibles à la maladie, de
même que les cultures
de tissus ont été
trouvés à la fois efficaces et économiques.
are fowl plague, avian encephalomyelitis, infectious
bronchitis,
tracheitis,
avian
infectious
leucosis
laryngoduck
complex,
plague, Mycoplasma gallisepticum, pasteurellosis
and ornithosis
avian
(psittacosis).
Non-infectious diseases caused by ribo flavin
vitamin
deficiency,
E
deficiency,
toxic drugs and toxic plant seeds may also need to be differentiated.
disease from a num
Sweden and Australia, where the slaughter
been successful. However,
measures
Germany,
forms,
as in
England,
The Netherlands the
application
and
and
Wales, Western
of slaughter
policies has failed to eradicate the disease.
The world distribution of Newcastle dis ease and the control ft
de
un diagnostic
ses qui
Parmi les maladies infectieu lui ressemblent et dont il faut la
distinguer,
on
compte:
la peste aviaire aviaire,
gallisepticum),
aussi avoir Newcastle
la
{Myco
pasteurellose
(psittacose). On peut la maladie
à différencier
de maladies
non
causées par des carences de riboflavine de vitamine
E, par
de
infectieuses ou
des drogues ou par des
graines de plantes toxiques.
has
where countries
have been invaded by the mesogenic lentogenic
difficile
aviaire et l'ornithose
velogenic form of the disease has occurred,
Canada,
rendent
différentiel.
plasma
ber of countries. In South Africa, England,
by
Newcastle
et de
la maladie
accompagnent
peste du canard, la mycoplasmose
Slaughter measures have been applied to
eradication
grande variété de symptômes
la bronchite infectieuse, la laryngotrachéite la infectieuse, le complexe leucémique,
Slaughter Measures and International Trade
Wales,
La
lésions qui
(fowl plague), l'encéphalomyélite
Control Measures
eradicate Newcastle
Diagnostic Différentiel
policies established by
Mesures de Répression Abattage Obligatoire et Commerce International On
a eu recours
pour l'éradication
à l'abattage obligatoire de la maladie de New
castle en certains pays. Un tel programme a réussi en Afrique du Sud, en Angleterre,
different countries have an appreciable influence on the international movement of live poultry, hatching eggs and poultry carcasses. In many countries in which Newcastle disease has a variable distribu of poultry and poul
tion, the importation try products
is very strictly controlled;
in countries Zealand, exist and
such as Australia and New
where Newcastle
at present, dead
tent
of
disease does not
and
prohibited.
hatching
eggs
is
To reduce the ex it
restrictions,
these
of live
the importation
poultry
completely
and
has
been
suggested that control measures be applied on the basis of "infected region of a
country"
of "infected
instead
au Pays de Galles, tralie
où
maladie
la
terre, Pays de Galles,
La répartition de Newcastle
disease
84 countries in 1962.
Compulsory notification of outbreaks of Newcastle
disease is difficult
partly because the disease
to enforce, is
often
not
in its lentogenic and asympto matic forms and is not, therefore, reported to the authorities. The existence of asymp recognized
mondiale
de la maladie
pays ont une in
établies par différents fluence
Pays-Bas,
et les mesures de répression
appréciable
international
sur
le
volailles
des
mouvement et
vivantes
abattues ainsi que des oeufs d'incubation. Plusieurs pays où la présence de la maladie de Newcastle
varie
et
It was notifiable in
Canada,
de l'Ouest.
Allemagne
Newcastle
the world.
l'abattage
obligatoire n'a pas réussi à réprimer les formes mésogénique et lentogénique de la maladie, dans les pays suivants: Angle
rigoureusement
disease be a notifiable
la
de
vélogénique
Toutefois,
sévissait.
country." Implementation of the concept of "infect ed region of a country" might require that throughout
en Suède et en Aus
forme
régissent
beaucoup,
l'importation de la volaille Certains pays indemnes
et de ses produits.
Nouvelle-Zélande l'Australie, ont complètement prohibé
de la maladie, comme la les
importations
abattues
de volailles
vivantes
d'oeufs
d'incubation. réduire la portée de ces restrictions, et
suggéré
mesures
que des
ou
Pour on a
de répression
soient prises sur une base de «région in fectée dans un pays» plutôt que celle d'un «pays
infecté».
Il
pourrait résulter
de
du concept d'une «région in fectée dans un pays» que la maladie de l'acceptation
tomatic outbreaks, and the use of live virus vaccines which can spread the infection to
Newcastle
susceptible
gatoire dans tous les pays du monde. Elle
poultry,
add to the problems
faced by national disease control agencies. Despite
these obvious
difficulties,
regions
of several countries have been freed from Newcastle disease for extended periods through control by slaughter.
number
of different
methods
tween make.
different
direct studies
comparisons are
difficult
L'obligation de déclarer
obli
les manifesta
tions de la maladie de Newcastle facile à mettre en application formes
lentogénique
et
n'est pas
parce que la
maladie est difficile à reconnaître
matiques have
of disinfectants on Newcastle disease virus. Some of the findings are tabulated in this though
con
l'était dans 84 pays en 1962.
autorités. L'existence
been used to evaluate the viricidal effect
review,
maladie
une
et à déclaration
dans ses
asymptomatique.
De ce fait, on néglige d'en informer les
The Action of Chemicals on Newcastle Disease Virus A
devienne
tagieuse «nommée»
be
to
vivants parmi
et l'emploi
de formes asymptodes vaccins
qui peuvent propager les
volailles
quent les problèmes
prédisposées,
pour
à virus
la maladie
compli
les organismes
nationaux chargés de sa répression. Toute fois, certaines régions dans plusieurs ont été exemptes de la maladie
pays
de New
castle pour de longues périodes grâce à la répression
par l'abattage. 7
Less Common Methods of Control
Action des Produits Chimiques sur le Virus de la Maladie
Hyperimmune serum, serum-virus
and
degrees of success, and experi
varying
de Newcastle
treatments have been used with
medicinal
mental results so far obtained
have been
Des méthodes
variées ont été utilisées
pour estimer l'effet
des désinfectants
le virus de la maladie
contre
La
de Newcastle.
inconclusive.
présente publication contient des résumés de ces données; cependant, les comparai
Control by Vaccination
difficiles à établir.
sons directes
entre
études sont
diverses
Vaccination has been the most widely adopted method of controlling Newcastle
In
disease.
1961, over 82 per cent of the
countries reporting vaccination
the disease were using
as their
main
control
proce
dure. The following immunological aspects are discussed
in
this
plurality; procedures and
munity;
factors
review:
antigenic
for evaluating
im
that
the
influence
of immunity,
including pas sive immunity, age at time of vaccination, vaccine virus titres, viral interference, dif ferences between individual birds and development
Vaccines are of two main types: live and
It
inactivated.
is convenient
into lentogenic
vaccines
to divide live
and
mesogenic
virus strains. Among the most commonly used lentogenic LaSota;
Bl, F
strains are the
and
the Kom-
among the mesogenic:
Mukteswar, Roakin and MK107.
arov,
Another type of live vaccine developed
by the attenuation
has been
of virulent
strains of virus and propagation
A
L'inoculation
rares de répression
de sérum
hyperimmun,
mixte (serum-virus) et les traitements médicaux ont été utilisés avec l'immunisation
plus ou moins
de succès, et les résultats
des expériences n'ont pas été concluants.
Répression par la Vaccination La vaccination universellement
a été la méthode
la plus
adoptée. En 1961, plus de
82 p. 100 des pays qui signalaient sence de la maladie utilisaient
to another disease.
susceptibility
Méthodes plus
la pré
la vaccina
tion comme principal moyen
de répression.
Les aspects immunologiques
qui suivent
sont étudiés dans la présente publication: pluralité
des
antigènes;
procédés
estimer le degré d'immunité
pour
et les facteurs
qui influent sur le développement
de l'im
munité: immunité de
la
passive, âge au moment vaccination, titrage des vaccins,
interférence
virale,
différences
Il
in mam
existe
deux
individuel
à une autre maladie.
les et prédisposition
sortes
de
vaccins:
des
vaccins
vivants
authors have described the clinical effects
vaccins
vivants
of vaccination
virus de lignées lentogéniques et mésogéni-
tissue cell cultures.
number
of im munity with each of these strains. In this review
their
and the duration
findings
are compared
and
discussed.
A to
wide variety of agents has been used
produce
inactivated
agents include lin,
crystal
ation.
A
vaccines.
beta-propiolactone,
violet,
and ultraviolet
considerable
These
forma irradi
amount of data has
et
vaccins
tués. Les
of
malian
des
sont
préparés
ques. Parmi les lignées plus communément
avec
lentogéniques
des
le
employées, on compte
Bl, F et LaSota, parmi les mésogéniques, les Komarov, Mukteswar, Roakin et les
MK107. Un autre type de vaccin vivant se prépare en atténuant certaines lignées de virus et en les propageant dans des cultures de cellules provenant
fères. Certains
de tissus de mammi
auteurs ont décrit les effets
been accumulated
It
of im
on the duration
munity produced
vaccines.
by inactivated
also, to consider
is important,
com
bined vaccines administered as a single inoculum, and the administration of two vaccines
but by different
simultaneously
routes.
The choice of the most suitable vaccine to meet any particular set of conditions very complex.
A
comparison
of immunity
tion
engendered
is
of the dura by Bl, F,
de la vaccination
cliniques l'immunité
lignées; on trouvera
lication
et la durée de chacune
que procure
Pour produire
des vaccins tués on utilise
tions
les rayons
par
accumulé
une
données sur la durée de l'immunité confèrent
Il
les vaccins
est intéressant
l'inoculation
d'étudier
and
part of any eradication feature
scheme. Another
that has become
one or more revaccina
need to conduct
if
is the
apparent
from one day of age through to the end of the laying year is to tions
protection
combinés
en une seule opéra simultanée
de deux vaccins par voies différentes.
Il
of high
would be preferable to live would form an essential
antigenicity vaccines
que
tués.
tion ainsi que l'administration
is
a
de
considérable
vated vaccines have resulted in very satis There
On
ultra-violets.
quantité
de vaccins
of immunity.
se
à leurs conclusions.
une grande variété d'agents: bêta-propiolactone, formaline, cristal violet et irradia
formalin-inactivated and BPL-inactivated
factory duration
ces
des détails et des comparaisons
rapportant
strains, indicates that, to date, BPL-inacti
little doubt that a killed vaccine
de
dans la présente pub
est assez difficile
conditions
choix du
de faire le
vaccin qui convienne
à toute une série de
Une
données.
comparaison que con
quant à la durée de l'immunité
Bl,
fèrent le
le
F,
le vaccin tué à la
BPL
line ou par le
a indiqué
présent le vaccin tué par le
forma-
que jusqu'a
BPL
donne des
résultats très satisfaisants.
Il
y a peu de
doute
à
antigénicité
vaccin
qu'un
tué
be achieved.
élevée,
As measures for the control of other avian virus diseases advance, there will be
vivants et entrerait dans les cadres de tout
an increasing
for live Newcastle
demand
disease vaccines prepared from tissue cul tures, preferably
of non-avian origin. One
modified Newcastle disease virus developed in
tissue
culture
is
distinctive
because
this virus has shown little or no tendency to spread and the immunity it engenders is
of considerable duration.
individually.
For
many
would entail abandoning methods. would
It
is doubtful
to each chicken countries,
this
mass vaccination
if
this
change
be made voluntarily. Government
assistance or legislation sary. Nevertheless,
d'éradication.
programme
qu'on doit revacciner d'assurer
vaccins
aux
préférable
Il
évident
est
une fois ou plus afin
une protection
depuis
la nais
sance jusqu'à la fin de l'année de ponte.
Au fur
et à mesure
méthodes
les
que progresseront
répression
de
autres
des
maladies virales des volailles, il se produira un accroissement vivant
de la demande de vaccin de Newcastle,
la maladie
contre
préparé de cultures sur tissus, préférable-
In the control of Newcastle disease by vaccination, a major issue is whether the poultry industry will accept the advantages to be gained by using vaccines which have to be administered
serait
might
be
neces
there is now evidence
ment d'origine points
vue,
de
aviaire. A plusieurs le virus modifié de la
non
maladie
de Newcastle
cultures
de tissus se distingue
parce qu'il ne montre tendance
à
se
développé
sur des autres
des
à peu près aucune
propager.
De plus, l'im
munité qu'il confère dure longtemps.
L'une qui
se
maladie est
des questions les plus importantes
posent
dans la répression
de Newcastle
celle de
savoir
de la
par la vaccination
si l'industrie
avicole
acceptera les avantages résultant de l'utili 9
that,
in general,
requiring in better im
vaccines
dividual administration
give
The development
munity.
of procedures of inactivated
for the mass administration or non-spreading tissue culture
vaccines
could lead to great changes in the global picture of Newcastle
disease.
The world history of Newcastle
disease
has shown that, once the disease has be in a country
come established
it tends to become has
happened,
made towards
endemic.
little
emergency
entraîner
de grands
problème
global de la maladie.
has sometimes the cost of
the vaccine has been subsidized.
of the literature
that no one vaccine
tués et non infectieux,
the use
value of the stock and the
review
indicates
has proved
changements
L'histoire mondiale a
Newcastle
que
methods
of use best suited to individual
It
is axiomatic
need to be supported
that these studies
by adequate labor
atory facilities.
In
any
discussion
against Newcastle
on
immunization
disease, it is necessary
to establish whether the protection is
against
whether
mortality
and
the greater
need
sought
paralysis; is
or
to protect
against the adverse effects of the disease on egg production and egg quality. 10
la
région, elle a tendance à devenir endémi que. Là où la chose s'est produite, on a
pour l'éradication
réalisé peu de progrès
de répression ont toujours d'urgence
Les mesures
de cette maladie.
éventuelle
eu un caractère
l'utilisation
et comprenaient
de
virus vivants et la vaccination
collective.
Le coût de la vaccination
est habituelle
avicole. Aux Etats-Unis, la vaccination des poulets à griller produits en 1956 a coûté plus de quatre millions de dollars. En Afrique, on
ment supporté par l'industrie
trouve
sont de vaccination hors de proportion avec la
que les frais
généralement valeur
countries.
de
maladie s'est établie dans un pays ou une
mande pour le vaccin a été faible.
for more government-sponsored
dans le
lorsque
areas. As a result, there would appear to experiments to determine the vaccines and
prove
de la maladie
démontré
ideal for all situations and all geographical be a need
de
collective
of an
been adopted in spite of serious difficulties.
A
découverte
de vaccins
demand for vaccine has been poor.
clearly
La
nant de cultures faites sur tissus, pourrait
are usually
the difficulties,
assurent une
individuellement
administrer
been
of the United States broiler crop in 1956 cost more than four million dollars. In Africa, the cost of vaccination has gener ally been considered out of proportion to
To overcome
pourrait être néces il semble évident main tenant qu'en général les vaccins qu'il faut
saire. Néanmoins,
Con
borne by the poultry industry. Vaccination
Compulsory vaccination
est
tion gouvernementale
pour l'administration
of live virus and mass vaccination.
the economic
Il
douteux qu'un tel changement soit adopté volontairement; une aide ou une législa
méthodes
be
à
qui
pays, l'aban
this
has
ce
en masse.
immunité.
nature and to involve
The costs of vaccination
de la vaccination
meilleure
eventual eradication.
trol measures have tended to
pour plusieurs
nécessiterait don
qu'il faut administrer
individuellement,
oiseau
chaque
or region, Where
progress
sation de vaccins
économique
La vaccination
sujets
des
obligatoire
a
et
la de
parfois
été
adoptée en dépit de difficultés sérieuses. Le coût du vaccin a alors été subventionné pour triompher L'examen
des difficultés.
des documents
n'existe aucun vaccin
indique qu'il idéal pour tous les
cas et dans toutes les régions ques. Aussi semble-t-il ments
devraient
afin de déterminer
géographi
que les gouverne
poursuivre
des
études,
quels sont les vaccins
et les méthodes d'emploi
les mieux adaptés
pays. Ces études exigeront laboratoires convenablement outillés. à chaque
Dans
tout
travail
sur
des
l'immunisation
Virus Propagation and Vaccine Preparation Newcastle
disease virus is classified
as
a Myxovirus and has been given the name
Myxovirus multiforme. The properties of the virus and the practical aspects of
and chemical properties.
been made,
Electron
studies of the virus have also and the arrangement
of its
structural components suggested. The virus has proved adaptable to a number of different living tissue cells of avian and mammalian
hosts and embryos.
The ease with which Newcastle virus can be propagated
is utilized
disease in the
and use of a variety
large-scale preparation
of vaccines.
At
time,
the majority of
Newcastle disease vaccines is prepared from virus propagated in embryonating chicken eggs. The main disadvantage to using eggs is the risk of their containing
of infectious
this problem,
a
agents. To overcome
vaccine strains of virus have
been propagated in tissue culture. The vast
majority of live vaccines is now dispensed as a lyophilized product and this has greatly
facilitated
transportation
and
storage.
A
la production
et la qualité
des oeufs.
Propagation du Virus et Préparation du Vaccin Le virus de la maladie de Newcastle est les Myxovirus et on lui a donné le nom de Myxovirus multiforme. On a beaucoup étudié les propriétés de ce
classé parmi
virus
aspects pratiques
et les
duction
du vaccin.
de la pro
Différentes
lignées du
virus ont été identifiées en tenant compte de leur
et de leurs propriétés
pathogénie
Des études du virus faites au
chimiques.
ont
électronique
microscope
suggérer la disposition
permis
structurale
de
de ses
On a trouvé que le virus peut s'adapter à un certain
bryons.
de tissus cellulaires
nombre
vivants d'oiseux,
de multiplication est
Sa facilité
utilisée pour la préparation échelle et l'utilisation tous
Presque maladie
et d'em
de mammifères
sur une vaste
de vaccins variés.
les
de Newcastle
contre
la
sont préparés
de
vaccins
virus propagés dans des embryons
d'oeufs
de poule, mais avec ce procédé on court le risque que les oeufs contiennent
Pour éliminer
d'infection.
des agents
ce danger,
on
propage des lignées de virus pour vaccin
number of procedures
standardizing
Newcastle
for testing and
disease vaccines
have been devised. However, at the present time, tests and criteria potency
est
constituants.
the present
variety
est
est de
sie ou si le besoin le plus important
have been identified on the basis of patho microscope
il
la protection
si
protéger contre les effets de la maladie sur
extensively. genicity
de Newcastle,
d'établir
recherchée contre la mortalité et la paraly
production have been studied Different strains of the virus
vaccine
la maladie
contre
nécessaire
of
vaccines
throughout the world.
for the safety and are not uniform
sur des cultures de tissus. On a mis au point certaines de vérification
et
méthodes
de normalisation
des
vaccins. Toutefois, à l'heure actuelle, les méthodes de vérification et les normes de sécurité et d'efficacité ques dans majorité
le monde
des
vaccins
tenant distribués
ne sont pas identi entier. vivants
La grande sont main
comme produits
lyophi
lisés.
11
DEFINITION OF NEWCASTLE Nomenclature
castle disease virus into two distinct groups
Newcastle disease takes its name from the town of Newcastle-on-Tyne in the of Northumberland, England, county where an acute disease occurred in a flock of poultry in the spring of 1926 (Doyle, 1927). In the following 15 years, the dis ease
was given more than 20 synonyms.
Many of these are listed in the publications of Reis and Nobrega (1956) and Beaudette (1943). The following are among the names
that
have
been
most
commonly
used: Ranikhet disease (Edwards,
1928),
(Picard, 1928; Hutra et al., 1938), pseudo-poultry plague (Johns tone, 1931), Doyle's disease (Haddow, pseudo-fowl
pest
1938), avian pneumoencephalitis (Beach, 1943), respiratory nervous disorder (Bang, 1946) and avian or fowl pest (gefliigelpest)
(Farinas, 1930; Kuppuswamy, 1935). Doyle (1935) thought the name New castle disease unsuitable;
nevertheless he felt that the terms "pseudo-plague" and "avian pseudo-pest" should be avoided be cause of possible confusion with another disease generally known as fowl plague. The name fowl pest was used in the
Fowl Pest Order of 1936, made under the Diseases of Animals Act of the United Kingdom of Great Britain and Northern Ireland. In British legal context fowl pest still applies to the two separate diseases, Newcastle disease and fowl plague.
Properties of the Virus The physical
of Newcastle disease virus have been reviewed by Elford et al. (1948) and a number of properties have been used to type different strains of the virus (Acocella, 1955; Anon., 1959; Hanson Brandly,
et al.,
properties
1949, 1951; Hanson
and
1955; Kaschula, 1952a; Upton
By means of pathogenicity and serological studies, MacPherson and Swain (1 956) divided eight strains of New
et al., 1953b).
DISEASE
—
both groups shared common
and
had
similar
reproduction
antigens patterns.
Other tests that have been used for strain identification
include
the
intracerebral
of mice (Nitzschke and Schmittdiel, 1960; Upton et al., 1953a, 1955) and the adsorption of haemagglutinating activity by suspensions of chick embryo brain cells (Piraino and Hanson, 1960). Piraino and Hanson (1960) have suggest ed that the pathogenicity of different strains of virus for chickens is related partly to the avidity of the virus for brain cells. The resistance of haemagglutinin to heating to 56°C has also been used to dis inoculation
between
tinguish
different
and
Schmittdiel,
microscope
examination
schke
strains
(Nitz
1963). Electron of a number of
strains of virus has revealed no
different
differences be appreciable morphological tween strains (MacPherson, 1956a; Muss1962; Reagan et al., gay and Weibel, 1948b, 1950b).
The general problem of determining
the
particle size of Newcastle disease virus has been investigated by Elford et al. (1948). In normal preparations, the average par ticle size is 1 92 microns. However, Reagan et al.
(1948) have reported virus particles
between 1 00 and 1 25 microns in diameter.
According to Cunha et al. (1947) the approximate size of the head piece is in the range of 80 to 120 microns (Burnet and Ferry, 1934). Schafer et al. (1949) and Reagan et al. (1950b, 1956) examined particles of New castle disease virus with an electron micro scope and reported that in aqueous solu tion they appeared to be rounded; whereas in saline solution they appeared thread-like in shape. This change in shape was con by Angulo (1951), (1948) and Schafer et al. (1949). Bang (1948), on the other hand,
sidered
Elford
an
artifact
et al.
considered
the filamentous
particles to be 13
SCHEME OF THE STRUCTURE
OF NEWCASTLE
DISEASE VIRUS
(Modified from SchciUr, 1963)
150
(ENVELOPE)
1
Figure 1.
individual virus particles. This latter view
gested that the virus particle
was based on the fact that the characteris-
sperm-shaped.
The
is essentially
filamentous
were not seen in
other virus preparations;
that the particles
in saline solution revert to the spherical form when
were agglutinated by specific antisera; and
replaced in water. These changes have not
in embryos,
that they produced
infection
Similarly, Cunha
al (1947)
14
et
have sug-
which occur when the virus
forms
particles
tic filamentous
been (Bang,
related 1948,
to
any
1949).
is
loss of infectivity The conversion to
Figure 2. — Electron micrograph of Newcastle disease virus (x 150,000). (Courtesy of Dr. A. P. Waterson, St. Thomas's Hospital, London.)
the filamentous
forms can be prevented by with formalin (Bang,
inactivation
partial 1947).
All
of Newcastle
strains
disease virus
studied by McCollum and Brandly (1955b) possessed mucinase. It has been suggested by Mierzejewski (1962) that the catalytic activity of aldolase in the breakdown of fructose
varies between dif
diphosphate
strains of the virus. and sero Using three neutralization logical tests, Doll et al. (1956) demon
ferent pathogenic
strated
that
serologically
Newcastle
disease
virus
is
and immunologically distinct
This virus density apparently differs ac cording to the cell type used for propaga tion. Thus, the infectivity peak densities of virus in avian cell culture have been lower than the corresponding figures for virus of mammalian cell origin. The ribonucleic acid (RNA) fraction of Newcastle
disease virus
has been found
for chick embryos, and there has been no proof that RNA is responsible for infectivity or agglutinins (Benedict et al., 1960). However, the use of lipids apparently has maintained the infectivity of the RNA from an Indian strain of virus non-infective
and swine influenza. Based on morphology,
(Dhar et al, 1963). The inactivation of Newcastle
there appears
be
virus has been examined by radiation
the
Wilson and Pollard (1958) have estimated that the total radius of the virus is at least
from the viruses of mumps, and of human
tween
to be no resemblance
Newcastle
viruses of influenza
In
caesium
Newcastle
disease virus
(Cunha
chloride
et
density
and
al,
1947). gradients,
disease virus appears to contain
with a wide range of densities (Stenback and Durand, 1963). infectious
particles
disease and
510a.
According to Cunha virus
has an internal
resembles
the
et al.
(1947), the
differentiation
analogous
that
morphological 15
structure of living cells. Negative electron microscopy ment
with ether has indicated
internal helical ribonucleo-protein between
tinguishable Newcastle
contrast
before and after treat
various
disease virus
that the is indis strains
(Waterson
Cruickshank, 1963). The significance the virus by Schafer
components
and
of
has been reviewed
(1963); and an explanation of
the principles
underlying their structure
and of the arrangement
16
of
of the RNA in
particular
(Valentine and Isaacs, 1957) (Figures 1 and 2).
has been suggested
The properdin system can inactivate Newcastle disease virus (Wedgwood et al., This inhibition requires all the of the properdin sys tem (Karzon and Bussell, 1960). As a result, a theory of resistance to infection with Newcastle disease virus which is not im part of the classical antigen-antibody 1956). known
constituents
mune concept has been formulated zon, 1956).
(Kar
PART
Spread of the Disease
I:
GEOGRAPHIC Early Reports of Diseases Resembling Newcastle Disease
A
Between
disease
to have been carried
to the
African
continent
infected
became
in the mid-nineteenth
considered
in common
ninger,
At
1949). the end of the century
there was
of New Mac-
probably an extensive epizootic
of fowl
of the world (Jacotof, 1950). These dis eases
in 1833 (Man-
forms
plague were reported from several regions
(Da Camara and Valadao, 1950; Castro Amaro, 1 964) . A disease suggestive of Newcastle disease was also observed ports
near Budapest
disease
1909 and 1915, diseases now to be atypical
century by small ships trading with Asiatic
by Petenyi
with Newcastle
during the 1949 poultry epizootic.
disease resembling Newcastle
believed
is
DISTRIBUTION
appear to have had characteristics
with Newcastle
disease and
were called "peste atypique,"
or "pseudo-
pest," or were given names with graphical
qualification
a geo
such as "Egyptian
pest" and "Madras pest." In March 1 926, Newcastle disease broke out in the Dutch East Indies near the towns
carbo) led MacPherson to believe that the cormorant was the source of in
of Batavia, Cheribon and Soerabaia, on the island of Java, where it exhibited an ability to spread quite unlike that of any known disease of poultry. The disease was first reported by Kraneveld (1926). Later Picard (1928) gave details of the first reports received from the Civil Veterinary Department of the Dutch East Indies and confirmed that in March 1 926 the Veterin at Buitenzorg, ary Institute Java, was aware of the existence of a serious poultry disease near the town of Batavia (now Jakarta). The Civil Veterinary Depart
fection in Scotland
in 1897, and again in
ment reported that the disease had caused
He has suggested
enormous losses in a number of districts on
castle disease in northwest
Scotland.
Pherson
(1956b) reached this conclusion after reading a Gaelic poem written in and doing research in the Hebrides.
1898
He has drawn attention to the fact that in 1 899 and 1900 the Scottish Congested
Dis
tricts Board distributed
hatching eggs and fowls to the same areas in which outbreaks
of Newcastle
disease
occurred
between
1951. Field data and experi
and
1949
ments with living cormorants
(Phalacro-
corax
the
1949-51
epizootic.
that cormorants
and other
sea
birds
are
infection and not mechanical carriers. He has also suggested
true
reservoirs
that
there
biological
of
be
may
a
long-established
adaptation to Newcastle
virus in the cormorant,
disease
and that the domes
tic fowl may be only a secondary However, this
Blaxland
explanation
the cormorants
host.
(1951) has rejected
and has maintained
that
around the Scottish coasts
the island and had also been identified remote areas throughout
in
the archipelago.
In several native villages (kampongs) not a single fowl was left. It was impossible to ascertain the origin of the disease. For Doyle (1948) also, the origin of the out breaks in the Dutch East Indies remained a mystery, but he thought it unlikely that the disease had existed as a latent infection among
the
native
fowls of the islands 1"
because these fowls proved
highly
the disease continued
sus
First Outbreaks of Newcastle Disease in England
Thereafter
The outbreak in a flock of chickens in Newcastle-on-Tyne in England in 1926 (Doyle, 1927), which was responsible for the name Newcastle
disease, was thought
with infected food from a ship (Anon., 1962). It is interesting to note, however, that the original flock
to have been introduced
involved ration
at Newcastle
supplemented
had also been fed a
with offal collected
from the town. There were two other out breaks diagnosed in England
in 1926: one
in Somerset, and one involving a number of farms in Staffordshire. The latter was thought to have resulted from the sale of
in a local
The
infected
birds
mortality
in all the 1926 outbreaks in Eng
market.
land was 98 to 100 per cent and, as there were
few
survivors,
to spread, reaching
Japan (Nakamura et al., 1933), southern India (Kylasamaier, 1931) and Australia (Johnstone, 1931) by the end of 1930.
ceptible.
the disease quickly
spread was increasingly
rapid
(Dobson, 1952). It has been noted MacPherson (1956b) that many of initial outbreaks occurred at seaports were associated with the movement
by the or
of
ships.
India In India, Newcastle disease was first re at the town of Ranikhet in the United Provinces in July 1927. Almost at the same time, a similar disease appeared in Madras State. As in the Dutch East Indies, the disease spread rapidly. By the end of 1927 it had spread throughout the United Provinces; by the spring of 1928 it had reached the Punjab and Bombay. cognized
Soon afterwards
the disease was reported
from all parts of India (Haddow, 1941).
died out. These English outbreaks were regarded by Doyle (1948) as chance offshoots of the main stream of infection that origin ated in the Dutch East Indies. the epidemiology
However,
of the early outbreaks
in England (Dobson, 1952) was very dif ferent from that experienced in the Dutch East
Indies
where
the
disease
spread
the whole territory within of six months (Picard, 1928).
throughout period
a
Spread Through Southeast Asia In
the
autumn
of
1926,
Newcastle
disease was reported in Korea (Konno et 1929; Ochi and Hashimoto, 1929).
al,
occurred at During 1927, outbreaks Ranikhet in the Kumaon Hills of India (Edwards, 1928; Cooper, 1930), at Col ombo in Ceylon (Sturgess, 1928; Craw ford, 1931) and at Manila in the Philip pine Islands (Farinas, 1930). Thus, in 1926
and
1927
Newcastle
disease
was
identified of Southeast Asia. From these known foci of infection in five countries
IS
Philippines By comparison Newcastle
with the rapid spread of
disease in some countries,
rate
of spread was slow in the Philippines. This was due, perhaps, quarantine
to the enforcement
regulations
of
and the movement
of poultry inward, toward the original focus of infection, rather than outward, toward the provinces (Coronel, 1939). An account of the spread of Newcastle disease in the Philippines during the tenyear period
1927-37
has been given
by
Coronel (1939). The disease spread in all directions from the original focus in the city of Manila. It usually made its first appearance in a province either in the capital of the province
or in
a large town
on a rail or road route, but it rarely peared in remote districts (barrios).
ap
Malaya In the province of Wellesley in Malaya, diseases, including Newcastle
poultry
the
Newcastle
of poultry dealers. The baskets in which poultry were carried permitted the droppings and discharges from in fected birds to fall on roads and paths between villages (Kuppuswamy, 1935).
page 84.
disease, were
through
spread mainly
activities
Haiti and Madagascar As in the Philippines, in both Haiti (Bush, 1954) and Madagascar (Buck, Newcastle disease first spread 1947) along main traffic routes to market areas.
United States In the United States, the appearance circa 1935 of a mild respiratory-nervous disorder of chickens in California, term ed "avian pneumoencephalitis" (Beach, 1942),
was not
identified
as
disease
until
(Beach,
1944, 1946; Brandly et
nine
years
Newcastle
had
elapsed 1944;
al,
1942). The disease reached the coast of the United States by 1944
Stover, east
(Cunha et al, 1947). By August 1946 it had been diagnosed with certainly in 17 including a number on or near the Atlantic coast (Brandly et al., 1946e; Bruner et al, 1947; Morgan, 1946; states,
Stubbs, 1946).
Canada Following the identification and subse eradication of Newcastle disease from the province of Alberta in 1950, a serological survey was made of 1 per cent of the total poultry population of that province. Only one suspicious flock was identified. The natural isolation of farm flocks in Alberta was probably responsible in part for the successful eradication of the quent
disease
Newcastle
an extensive
disease
made in Canada. eight
and Bigland,
(Ballantyne
During 1952/53
provinces
virus Some
were
results of serological cent
to
1954a).
1951). survey of
was antibody 2,130 flocks in
sampled, tests
showed
and
the
12 per
(Crawley, Further details of the history of have
been
infected
disease in Canada are given on
Africa In Africa south of the Sahara, castle disease was recognized
New
at the ports
of Mombasa in 1935, Durban in 1944, Cape Town in 1949, and Leopoldville in 1948 (Vandemaele,
Figure
These
3.
1961),
as shown
caused by fresh introductions (Scott et
al,
in
probably of infection
were
cases
1 956) , although
evidence was
found that the disease had existed in enzootic form along the coasts of Kenya and Tanganyika for some time. A survey conducted in the Congo has shown that the disease there has been con fined
mainly
to small
flocks
owned
by
Africans (Depoux and Chambron, 1960). However, in the Union of South Africa outbreaks have involved a number of large poultry flocks (Anon., 1950b). The main epizootic pathways in Africa have been closely related to the main rail and road trade routes.
Spread Through Europe The incidence of Newcastle disease and fowl plague in Europe during the period 1940-55 has been summarized by Eckert (1957), as illustrated in Figure 4. The incidence of Newcastle disease in the same countries in 1962 is shown in Figure 5. In many countries, Newcastle disease has shown a marked tendency to variation This tendency is for in pathogenicity. mesogenic and lentogenic strains to replace the initial velogenic form of the disease. have been
Such epizootiological changes recognized
in western Germany (Fritzsche,
1963), France (Lissot, 1956), Yugoslavia
(Jaksic and Stefanovic, 1957), and Great Britain (Anon., 1962b). In some countries epizootiological
changes
fluenced by control
have
been
in
policies. Further men
tion of this topic is made under the head ing "General Characteristics
of the Dis
ease" on page 45. 19
NEWCASTLE DISEASE IN AFRICA SOUTH OF THE SAHARA Dates of First Outbreaks and Direction (Redrawn from Vandemaele
Figure 3.
-n
1961)
of Spread
N0I1D3JNI
A1ISN3Q dO 3H1SADM3N
3SA3SIO aNV HMOd 6i of , ss6i
3novid ni 3doan3 omana 3hi aoiaad
H3D3 JD»A J»d OOO'l uMDJpay)
prrrj
mOJf
*Je>Ic3
I** (ZS61
o*o ■ o*i
iz
— Outbreaks of Fowl Pest in Great Britain — Financial Years 1954/55 to 1960/61 (Anon., 1962b)
TABLE
1
Financial year
No. of adult
No. of
hens
chickens produced
Outbreaks
broiler
kept
(million)
Primary'
Total
Secondary2
(million) 550
1 954-55
52
20
162
388
1955-56 1 956-57 1957-58 1 958-59 1959-60 1 960-61
54
40 47
154
757
214
1,036
56 82 108
236 245
822 742 537
391
2,333
782 2,724
142
342
1,529
1,871
58 59 63 66 63
911 978
1 A primary outbreak is one which has no established connection with a previous outbreak. 2 A secondary outbreak is one where the infection is known to have spread from another outbreak.
Great Britain The spread of Newcastle disease in Britain has been well documented (Anon., 1962b; Asplin et al, 1949; Cal ender, 1958; Gordon, 1961; Reid, 1955, 1961). In 1950, the tracing of birds moved from a large poultry show in England dis Great
In
closed 255 outbreaks.
of
infected
hatchery breaks.
stock
growing
resulted
In the
in
1955, movement
108
from
secondary
one out
same year, illegal movement
of infected birds resulted in 187 outbreaks being confirmed
in various counties.
Out
breaks in broiler chickens
in 1960 con involving 375 premises. The rapidity of spread of New castle disease (fowl pest) in Great Britain and the difficulty of tracing sources of infection is illustrated in Table 1. tributed
to
an
epidemic
The origin of outbreaks disease which occurred though
infection
originated
sea birds
it is possible
of infection
that the
in an adjacent colony
(Anon., 1951-52).
outbreaks were confirmed
22
of Newcastle
in 1950 in south
Ireland has not been determined with
certainty,
of
two
additional
outbreaks
occurred
Newcastle
disease
on
adjacent farms.
Austria In
Austria,
appeared
During
of the disease in most
incidence reached
first
1942 (Grausgruber, 1963). the period from 1946 to 1961 in
1947 and
a peak between
areas 1951
and declined thereafter. In the Burgenland area of the country, occurred
extensive outbreaks
in 1954, 1955 and 1957.
World Distribution In 1962,
103
in 1962
countries
reported
New
(Table 2). A number of countries, including Jordan, Mauritius, The Netherlands, Pakistan, Romania and castle disease
Ireland
east
ment of chickens and turkeys by three poultry dealers was responsible for the spread of the disease to 12 premises; and
Fourteen
and a sequence
was established.
The move
Venezuela, cidence.
reported
a decrease in its in
On the other
hand,
Argentina
(Anon., 1961a; FAO-WHO-OIE, 1962), Denmark (Marthedal et al., 1963) and (Vittoz, 1962), which had Luxemburg previously been free from the disease, re ported it for the first time. Iceland, Nor-
way, Sweden, Finland, the Republic of Ireland, Australia and New Zealand were
TABLE
2
among the countries
free from Newcastle
disease in 1962 (Vittoz, 1963).
— Incidence and Control of Newcastle Disease, 1962 (Lancaster, 1964b)
Countries reporting
Geographical area
rag
g 2
l| ll
h
of
Countries conducting
|
o
mo
.52
08
1=
|1
„
11
■s!
!§ o
ci
§p
fl
^..o
«{
i§
5
3
1
—
3
3
1
Central Africa
24
7
8
4
6
3
7
South Africa
22
4
9
9
16
6
7
South America
24
12
7
2
13
6
11
North America and West Europe
24
4
9
9
17
10
a 4
North Africa
East Europe
and Asia
21
6
12
1
19
15
South Asia
23
12
9
1
8
13
9
6
—
—
4
2
—
—
48 (53)'
55
Oceania Totals
149
1 The figures in parentheses
are for 1961
30(26)
84 (81)
56 (46)
45 (43)
(Lancaster, 1962a).
MODES OF SPREAD Newcastle disease has been known to spread in a great many different ways. The known sources of infection are discussed below
but
it
is conceivable
that
other
of spread exist and that they account for outbreaks of the disease whose origin it has not so far been possible to trace (Levine, 1952). modes
The source of an appreciable number of outbreaks has not been determined (Gordon etal, 1948; Levine, 1952; Mansjoer, 1961); and up to 1941 it had not been possible to trace the origin of any initial outbreak in a country (Haddow, 1941). It is particularly hard to trace the origin of the disease with certainty when 23
INCIDENCE
(Compiled
OF NEWCASTLE DISEASE IN EUROPE IN
from Animal
Health Yearbook
No Evidence
Low Sporadic Incidence Seasonal
Occurrence
Widespread Confined No Data
N.
IRELAND (
d
IRELAND
GREAT BRITAIN
Throughout
to Certain
the Country
Regions
FAO-WHO-OIE,
1962
1962)
occurs in a mild form (Levine, 1962a). In discussing the dissemination of New castle disease, Brandly (1950) has com mented on the low incidence of the carrier state in this disease as compared with human influenza in which, according to Burnet (1945), one carrier individual in
it
10,000 may suffice to initiate an epidemic.
The world history of Newcastle disease has shown that, in general, once the disease has become region,
established
in a country
or
it has tended to become endemic
(Doyle, 1948; Lancaster, 1963a). In areas with a dense poultry population, the har bouring of the virus in a few survivors, the resistance of the virus, and the existence of an appreciable number of potential hosts are enough to account for the continuing emergence of new cases (Asplin, 1961 ).
1960);
Cabassi, martin
and
house
European
(Delichon urbica) (Winmill
and
Weddell,
1961). Crows have
shown
of Newcastle
symptoms
(Haddow,
1941);
and
nervous
typical
disease in India they
have
been
found dead or dying in the vicinity of out
involving chickens in Indonesia (Picard, 1928), Ceylon (Crawford, 1931) and India (Cooper, 1930; Sahai, 1937a). The virus has not, however, been isolated from some crows (Crawford, 1931) and jackdaws (Keymer 1958, 1961) found dead or dying in the vicinity of other breaks
outbreaks.
The role of the sparrow (Passer domesticus) is difficult to assess. Gustafson and Moses (1953a) and Hartwigk and Nitsch (1957) found that fowls developed New castle disease after being placed in contact
with infected
Wild Birds
sparrows.
Contrary results
have been reported by Maglione (1956). Popovic (1951 ), and also Placidi and San-
Newcastle disease virus has a wide range of susceptible avian hosts. This has been demonstrated in both naturally occurring
tucci (1953a), found the sparrow was not susceptible to Newcastle disease virus; but
and
infections
this finding has not been supported by the
is import
observations
experimentally
produced
(Beach, 1946; Levine, 1952). It
of Pomeroy and Fenster(1948) and Kaschula (1950).
that the part played by wild birds be considered whenever the origin or
macher
means of spread of outbreaks
ported by Kee (1928). In recent years it has been unusual to find crows and spar
ant, therefore,
gated (Schoop et.
al,
is investi
1955).
Newcastle disease virus has been re covered from the following species of wild birds: starling (Sturnus vulgaris) (Gilles pie et al., 1950); gannet (Sula bassana) (Wilson, 1950); sparrow {Passer domes(Gustafson and Moses, 1953a; Keymer, 1961; Monda et al, 1960); shag (Phalacrocorax aristotelis) (Blaxland, 1951); grey parrot (Psittacus erithacus) and Winmill, 1960); jackdaw (Scott ticus)
Inconclusive
field evidence
rows dying in the vicinity of outbreaks of Newcastle
disease
(Parnaik
natural field infections have in volved parrots in Africa (Malbrant, 1942; Scott et al., 1956) and jackdaws land (Keymer, 1961).
Many different
species
birds have been infected
parakeet
a zoological
ochrocephala)
(Cavrini
and
in Eng
of free-flying artificially with
Newcastle disease virus (Table 3).
Infections in Birds in Zoological Gardens
(Amazona
Dixit,
and
1953). Other
(Corvus monedula) (Keymer, 1961); koel (Eudynamis scolopaceus) (Shah and John son, 1959); great horned owl (Bubo virginianus) (Ingalls et al., 1951); osprey 1952b); (Pandion haliaetus) (Zuydam, (Palaeornis) (Zuydam, 1952b); swan (Cygninae) (Vrtiak, 1958); parrot
has been re
In an outbreak of Newcastle disease in deaths
garden in Surabaja, Indonesia, among paradise birds,
occurred
pheasants,
lyre
birds
and
rice
birds 25
TABLE 3—Wild Birds Susceptible Experimentally to Newcastle Disease Virus and Methods of Evaluation (Modified from Gustafson and Moses, 1953b) Name
Vernacular
Scientific Bubulcus ibis Lophortyx Californica Colinus virginianus Perdix perdix Alectoris graeca Phasianus colchicus Gennaeus nycthemerus Francolineus capensis Columba livia
Methods of evaluation
Stilt bird California quail Bobwhite quail Hungarian partridge Chukar partridge Ring-neck pheasant Silver pheasant Cape pheasant Pigeon
V
Placidi and Santucci (1953a) Beach (1942)
Hl
| V HI
Geopelia striata
Columba palumbus Streptopelia turtur Streptopelia
Japanese dove. Stone pigeon. Striped ground dove Ringed pigeon Forest turtledove Ringed dove
Fenstermacher ef al. (1946) Beach (1942) Kaschula (1950) Beach (1942) Fenstermacher et al. (1946) Hanson and Sinha (1952) Placidi and Santucci (1953a) Collier and Dinger (1950) Mansjoer (1961) Kraneveld and Mansjoer (1950b) Martini and Kurjana (1950) Placidi and Santucci (1953a)
risoria
Streptopelia senegalensis Streptopelia chinensis
Author
Kaschula (1950)
Laughing dove
D, V
Spotted dove
S, D
Martini and Kurjana (1950)
Mourning dove Dove
S, D S, D
Jezierski (1950) Hanson and Sinha (1952)
tigrina
Zenaidura macroura Streptopelia (hybrids) Munia punctulata
Prit
misoria
Bondol Bondol
Munia maja zaperena Munia ferriginosa Uroloncha leucogastra leucogastroides Padda oryzivora Ploceus manyar Passer melanurus Passer domesticus
Glatik Manyar Cape sparrow English sparrow
S, V
Corvus monedula
Jackdaw
V.HI
Corvus brachyrhynchos
American crow
D, HI, V
S — symptoms D —death
Prit
S in some ; D and serial passages in last 2 species
D, V
HI — haemagglutination inhibition activity of serum V — virus isolation or virus transmission
26
Collier and Dinger (1950)
Kaschula (1950) Gustafson and Moses (1 953a) Mansjoer (1961) Baczynski (1960a) Keymer (1961) Karstad etal. (1959)
(Mansjoer, 1961). A similar outbreak in Morocco resulted in deaths among chick ens, pigeons and pheasants; whereas some predatory species and species belonging to the orders Passeriformes and Psittaciformes did not succumb (Placidi and Santucci, 1954). In one outbreak in a zoological garden in Germany (Kloppel, 1963; Schoop etal., 1955), the disease appeared in specimens of little owl (Athene noctura), raven (Bucorvus), eagle (Haliaetus albicilla) and kingfisher (Dacelo gigas) . Among the owls the disease was peracute, but in the other species the disease was more chronic and caused cerebral symptoms. Although a number of different species were exposed, it was noted that the Gallinaceous birds were not affected (Kloppel 1963; Schoop et
al,
1955).
A
second outbreak occurred
in the same zoological garden some 18 months after the first (Kauker and Siegert, 1957) and involved ostriches (Struthio camelus), a vulture (Pseudogyps Africanus Salvad) and a toucan (Ramphastos dicolorus). Newcastle disease has also been diagnosed in Germany in a king penguin (A ptenodytes patachonica ) ( Krauss et al., 1963).
In Italy, the infection was introduced to a zoological garden
by newly
imported
parrots (Amazona ochrocephala) and then spread to a peacock, a jungle fowl, a
(Ara severa) and a guinea-fowl (Aery Ilium volturinum) (Cavrini and macaw
In India,
Newcastle
of Newcastle
Zoological
Gardens,
disease appeared
in a
of Indian partridges that had re cently been obtained from a dealer (Parnaik and Dixit, 1953) and within four days all 35 birds died. number
been
(Psit-
1952b).
Migratory
birds
probably
introduced
disease virus to Cyprus (Crowther, 1952). On the sea coast of Britain, Newcastle
shags, gannets and cormorants considered
important
have been
agents in the spread
of Newcastle disease to poultry (Blaxland, 1951; MacPherson, 1956b; Reid, 1955; Wilson, 1950); and inland, jackdaws and starlings may have played a similar role (Keymer, 1961; Locke, 1960).
Excretion of Virus in Faeces The excretion of Newcastle disease virus in the faeces of several species of birds led Gillespie Moses
al.
et
(1953a,
(1950), Gustafson Karstad
1953b),
and et
al.
(1959), Seetharaman (1951b) and Spatalin and Karstad (1959) to believe that poultry farms become infected may through the faeces of birds. A similar opinion has been expressed by Callender (1958), and also by Hanson and Sinha (1952), although they did not have de finite evidence to support this view. Kaschula ( 1 950) has suggested that the faeces of wild birds may have played a part in the spread of Newcastle disease in South
Africa. by crows and hawks was
likely in India (Sahai, man, 1951b)
1937b; Seethara
and in Pakistan
(Khan and Huq, 1963). Haddow (1941 ) has reported that in India, fowls could be infected with virus from crows. However, Gustafson and Moses
(1952) failed to demonstrate of Newcastle disease from
infected sparrows to chickens by close association; even though the sparrow was
Imported Birds There is danger of spreading Newcastle disease when birds are transported by air from one country to another. For example, in both Scotland and The Netherlands
parakeets
Introduction of Virus by Migratory Birds
transmission
Introduction of Virus by
disease have
(Anon., 1958; Zuydam,
tacidae)
Dissemination
Cabassi,
1960). the Bombay
breaks
associated with imported
out
naturally susceptible to the virus. Further more, attempts to transmit a Malayan strain of Newcastle disease virus to a number of species of Malayan birds have 27
results (Orr and John,
given inconclusive 1946).
castle disease from Calcutta to the Nether
Game Birds Game
and pheasants in particu
birds,
lar, have been associated with outbreaks of Newcastle disease in poultry in Britain (Gillespie, 1952), France (Moine, 1950), Canada (Moynihan et al., 1951), Italy
(Brandly
al.,
et
1948),
(Wagener,
1946a),
Germany
Czechoslovakia
(Jera-
bek, 1961) and The Netherlands (Jansen and Kunst, 1952). It has not always been clear whether the game birds were infected to infection by poultry or introduced Wagener ( 1948 ) poultry. In Germany, thought that pheasants (Phasianidae) prob ably infected domestic poultry. A similar view was taken by Jerabek (1961) in However, Reid (1961) Czechoslovakia. believed that, in Britain, pheasants did not constitute a substantial reservoir of infec tion. Outbreaks pheasants
of
have
Newcastle also
there has been little
occurred
disease
in
in which
or no evidence of
association with domestic poultry (Levine 1949; Locke, et al., 1947; Liebengood, 1960; Skoda 1954; Vrtiak, Osthoff,
and Zuffa, 1958;
1956b; Torlone,
Weidenmuller and
1953; Zuydam,
The shipment of pheasants, partridges and other game birds has introduced New
1950a),
and in
lands (Jansen et al., 1949; Jansen and Kunst, 1952; Zuydam, 1949), from the mainland of the United States to Hawaii (Adler et al., 1951), from Hong Kong to California (Anon., 1950a), and from Spain to the United States (Thompson, 1955). In some of these incidents, birds were dead on arrival and this facilitated early diagnosis. In one instance, imported partridges (Per dix perdix) died while being held in a quarantine
station (Thompson,
1955).
with Newcastle disease virus in several species of game infections
Experimental
birds have been reported by Pomeroy Fenstermacher
and
(1948) and by the authors
listed in Table 3.
Pigeons and Doves The part played by domestic pigeons (Columba livia) in the spread of Newcastle disease has been examined by a number of authors. vations
Both laboratory have indicated
and field obser that pigeons
can
spread the disease.
Kaschula (1952c) reported that pigeons could excrete virus in the faeces for several days after being dosed with virulent virus. However, instances have been recorded
the mortality among outbreaks pheasants has been high (Levine et al., 1947; Uzieblo, 1961; Weidenmuller and
where infected pigeons have not spread the
(Popovic, 1951) unless the pigeons
had
there have 1954). Furthermore, been indications of Newcastle disease anti bodies in sera from pheasants raised on
been
and
some
Osthoff,
game
farms
etal,
1963).
(Andrews,
Natural infections
1963; Andrews
have been reported
in guinea-fowl (Numididae) (Crawford, 1930; 1930; Kretzer, 1931; Farinas,
Moine, 1950); in partridges (Perdix sp.) (Mantovani and Ceretto, 1953; Parnaik and Dixit, 1953; Torlone, 1954; Vrtiak, 1958); and in peacocks (Pavocristatus) (Jansen and Kunst, 1952). The partridge has been considered a likely carrier of the virus (Placidi and Santucci, 1953a). 28
disease to chickens infected
during
intranasally
cohabitation (Schyns
Florent, 1951). It has also been observed that pigeons and doves in contact with in fected chickens or contaminated premises have remained clinically healthy (Adler et al, 1951; Crawford, 1931; Kee, 1928; Orr and John, 1946). Reuss (1961a) has con cluded that pigeons under natural
condi
tions do not spread the virus from infected poultry
flocks. Reuss ( 1 96 1b) and Walker et al. ( 1 954a) found that pigeons infected per os excreted Newcastle
disease virus for at least three
days in sufficient chicks
placed
quantity in
contact
to infect baby with
them.
nected with the spread of infection
Pigeons placed in direct contact with New castle disease-infected a
sub-clinical infection.
turn,
transmitted
the
chickens
These pigeons, in disease
to young
Turkeys
chicks. Similar results with pigeons have by Reuss (1961a); and with doves (Streptopelia chinesis) by
been reported spotted
Kraneveld and Mansjoer (1950b). have Naturally occurring outbreaks been observed both in flocks of pigeons (Hanson and Sinha, 1952; Marastoni and Sidoli, 1959; Picard, 1928) and in in dividual pigeons (Iyer, 1939; Placidi and Santucci, 1953a; Vrtiak, 1958; Zoletto, 1958). Recovery of the virus from wild pigeons has been reported by Hanson and Sinha (1952) and by Vrtiak (1958).
Ducks and Geese Cases of naturally occurring Newcastle disease in ducks and geese have been re 1954; 1943; Bush, (Beaudette, ported
Moine,
1 950) but these species have been
as being more resistant to infec tion than fowls (Albiston and Gorrie, 1943; 1947; Beaudette, 1942; Asplin,
regarded
Berthelon and Tournut, 1949; Iyer, 1945; Kaschula et al, 1946). Artificially infected ducks and geese have been found
to excrete virus from
3
or 4 days (Asplin, 1947) to 15 days (Teklinskaer al., 1956).
Ducks and dissemination
of Newcastle
after infection
(Winmill and
that ducks and geese were con
a more
important
part
castle disease. Early reports of naturally occurring outbreaks in turkeys have been 1949a) and (1946). Recent re
reviewed by Beaudette (1943, Fenstermacher
et al.
ports include those of Buck (1947), Gale
Gordon (1954), Levine Walker (1948). et al. ( 1 96 1 ) , et al.
Gray (1947) and
et al. ( 1948 ) , et al.
turkeys from infect
Apparently-healthy
ed areas have been considered
responsible
for spreading Newcastle disease in Great Britain (Asplin et al., 1949; Gordon et al., 1948) and in the United States (Fenster macher et al., 1946). Asplin (1949) has suggested that some turkeys carriers.
Turkeys
believed
to have
may remain
imported
by
transmitted
air
are
a disease,
thought to be Newcastle disease, from Italy to Algeria (d'Arces, 1949; Donatien and Gayot, 1946). Furthermore, one in fected turkey that was kept in isolation for 12 months yielded virulent
Newcastle
dis
from the intestinal contents when it was killed (Winmill and Haig,
ease
virus
1961).
Chicken Eggs
disease virus.
Haig, 1961); and the observation that an outbreak of Newcastle disease followed of uncooked goose viscera the feeding (Heller, 1957). Similar conclusions have been reported from Pakistan by Khan and Huq (1963), and from Denmark by Marthedal et al. ( 1963 ) . However, in an outbreak in Cyprus there was no field evidence
Turkeys play
than ducks or geese in the spread of New
geese may play a part in the
This has been indicated by the isolation of virus from 48 out of a sample of 265 duck tissues (Vrtiak, 1958); the recovery of virus from the intestinal contents of ducks six months
(Crow-
ther, 1952).
developed
Newcastle disease vaccine virus has been recovered from the surface of the shells of eggs laid by vaccinated
after vaccination
birds for four days
(Zargar
and Pomeroy,
1950b). Virus was also recovered from the intestinal contents of these birds and contamination
may have come from this
source.
Examination of tissues taken from 283 birds flocks
in
105
Newcastle
has shown
disease-positive
20 out of 47 ovarian
yolks and 3 out of 10 oviduct egg yolks to yield the virus (Beaudette et al., 1948a). In addition, inflammatory changes in the 29
associated
ovary
infection
with Newcastle
disease
(Biswal and
have been reported
Morrill, 1954). A number of authors have recovered disease virus,
Newcastle
(either
vaccine
or field strains) from the contents of eggs laid two to ten days after vaccination of hens (Bivins et al, 1950; Hitchner et al, 1950; Prier et al, 1950; Van Waveren, 1955; Zargar and Pomeroy, 1950b). Three isolations were made from the yolks of 85 eggs gathered at the beginning of an out break in a laying flock (Thompson and Osteen, 1948). Eggs laid during the period was severely de when egg production
found
(1951a) embryos
that
majority
the
with
they inoculated
of
Newcastle
died by the ninth day of Virus was recovered from these embryos. No virus was recovered from embryos dying on the nineteenth day, from embryos failing to hatch, or from those chicks which did hatch (Doll et al, 1950e). Similar findings, but with eggs laid during natural outbreaks, were obtained by Hofstad (1949b) who con disease virus
incubation.
cluded that embryos that survived the first 1 0 days of incubation were apparently free
from Newcastle with
inoculated
disease virus.
Embryos
less pathogenic
strains of
disease virus have been known
pressed yielded
chicks from which virus four days after hatching (DeLay, 1947; Mansjoer, 1961); virus was also isolated from eggs laid during the Jungherr and Terrell recovery period. and Mansjoer (1961) demon (1946) strated the presence of virus in eggs laid
Newcastle
was
to hatch (Asplin, 1952; Rao and Agarwal,
two months after an active outbreak.
and the hatching of virus-infected
isolated
Field evidence
of Newcastle
disease
virus being passed through the egg to the chick has been reported (Sinkovics, 1957a;
Walker and Powell, 1950). From the information available to him, Beaudette concluded that virus — either (1948a) vaccine or field strains — is deposited in
1960).
Young Chicks disease virus on the surface
Newcastle
of the
egg,
the breaking
of infected
eggs,
chicks,
are all means whereby a hatchery can be
Newly hatched chicks from an infected hatchery have also spread Newcastle disease (Callender, 1958; Jung herr and Terrell, 1946). Furthermore, re
come contaminated.
ports
have
emphasized
the dangers
in
herent in the proximity of a commercial
only a relatively short period. Furthermore, Placidi and Santucci (1953c) found no evidence of egg transmission of
hatchery to a poultry dressing plant (Jung
the virus. Nevertheless,
brooder
eggs during
herr and Terrell, 1946) ; to a poultry rear ing farm (Reid, 1955, 1961); or to a
dried eggs
room (Schmittle and Mansfield, 1950) . In some outbreaks in young chicks, it has been impossible to determine
mercial eggs
whether infection
Newcastle
disease
virus is considered to have been present in
(Alegren, 1951) and in com (Tiefenbacher and Woernle, 1957) imported from a foreign country. No experimental evidence of permanent egg transmission was obtained by Asplin (1952), Bivins et al (1950), Hofstad (1949b) or Walker and Powell (1950); and Beaudette
(1948b) considered it safe laid
three weeks
to incubate
eggs
vaccination.
Although some embryos in
fected with virulent
(DeLay,
1947),
virus
after
have hatched
they usually
die during
incubation.
Doll 30
et
al
(1950e)
and Hitchner et
al
occurred in the hatchery
or during transit to the farm
(Jungherr,
al,
1950; Walker, 1948). Pomeroy and Fenstermacher ( 1 948 ) sus 1948; Olson et
pected infection Infected
of chicks whilst in transit.
day-old
have introduced
(Divo, 1950)
chicks
are thought
to
the disease to Venezuela and Hawaii
(Adler
et
al,
1951) .
Older chicks have played an important part in the spread of the disease in the United States (Schmittle and Mansfield, 1950;
Fenstermacher
et
al,
1946;
Pomeroy and Fenstermacher, 1948). Out of 148 outbreaks in young chicks reported 26 by Byerly (1948), 39 (approximately of the movement per cent) involved
(Kraneveld and Nasoetion,
The following discussion of the exist ence of Newcastle disease virus among growing and adult chickens is based on studies involving artificial exposure or vaccination.
Excretion of Virus from Respiratory System The spread of Newcastle
and
to susceptible
occurs
most
administered
chickens readily
the virus
to the donor group
is
by the
respiratory route. This indicates that trans mission is mainly by air-borne particles (Andrewes and Allison, 1961 ; Kohn, 1955; Miller and Miller, 1950; Robin, 1962; Schyns and Florent, 1951). In immune chickens, re-infection with virulent virus appears to be confined to the respiratory tract and lasts from 4 or 5 days (Doll et al, 1 950c; White et al, 1 954) to approximately 30 days (Zuydam, 1951b). In non-immune chicks, virus has been recovered from the respiratory tract from 6 days (Miller and Miller, 1950; White et al, 1954) to 21 days (Walker and McKercher, 1954b) after infection. There is as yet no evidence that the respiratory system acts as a permanent reservoir of the virus, although Beach ( 1942,
1951 ) ; and 5 weeks
(Van Waveren,
1 943 ) has claimed that adult birds,
1955;
1951b, 1952a). The passage of virus through the intestinal tract
Zuydam, virulent
of immunized fowls in this way apparently does not cause any detectable change in its pathogenicity (Dinter and Bakos, 1953) or virulence (Zebrowski, 1956). From studies of this type, involving have concluded
authors
placed in contact when
Brunner,
7 days (Krane 9 Mansjoer, 1950a); days 1952); 2 weeks (Woernle and 1957); 3 weeks (Adler et al,
either live or inactivated disease virus
1938). In pre excretion of
occur for varying periods:
(Asplin,
Growing and Adult Chickens
chickens,
or field virus has been found to
virulent veld
started chicks.
vaccinated
viously
infection
apparent
vaccines, several that mild
or in-
with virulent virus
is
possible in vaccinated flocks (Adler et al,, 1951; Asplin, 1952; Doll et al, 1951b;
Hofstad, 1953a; Zuydam, 1952a). Schmidt and Bindrich (1956) estimated that in some clinically immune chickens appreci able virus multiplication occurred in the organs which excreted virus. The excre tion of virus in the presence of circulating antibody has suggested local multiplication of the virus in the intestinal wall, un hindered
by
the
circulating
antibody
(Kohn, 1959). Although they did not determine the Beaudette exact means of transmission, etal. (1949) and Zuydam (1953) believed chickens vaccinated with a mesogenic strain could be a source of infection by contact for about three weeks following vaccination.
When chicks have been vaccinated with mesogenic
strains,
the
virus
has
been
recovered from faeces for 15 days, 19 days and 21 days (Van 1951b) 1955). Hitchner and Johnson (1948) found that the lentogenic vaccine strain Bl did not spread from vaccinated (Zuydam,
which recovered from the disease, carried virus in the lungs for three months.
Waveren,
Excretion of Virus in Faeces
chicks to other chicks in a separate com
During the early outbreaks of Newcastle disease in Indonesia, the virus was found in the faeces of chickens suffering from naturally occurring cases of the disease
of the same brooder.
partment this
for
Bl
Although
strain was excreted in the faeces
7 days
(Kohn and Ebert, 1960) to
days (Van Waveren,
1955),
14
direct physi 31
SUGGESTED SCHEME FOR THE PATHOGENESIS NEWCASTLE DISEASE VIRUS (Redrawn
from
Kohn,
OF
1959)
AIR
FOOD AND WATER
FAECES
MUCUS IN THROAT AN D NOSE
I
INGESTED
INHALED
INTESTINAL
UPPER RESP.
MUCOSA
MUCOSA
a
EtN FECTION
-L=s{ [VISCERA
BLOOD
H
SPLEEN
LUNGS
Figure 6. cal contact
was necessary
for spread to
by wire partitions kowski,
Contact
vaccinated
transmission
ceased two
weeks
of chicks with the (Hitchner and White, 1956). Baldelli (1956) concluded that, after oral administration, Strain F virus (Asplin,
has been reported
1957; Bankowski et
susceptible chicks (Bankowski et al., 1957; Marek, 1960; Markham et al., 1951).
al,
( Ban 1958a;
Bankowski and Corstvet,
1960). Nonchicks experimentally infected
after the re-vaccination
with field virus excreted virus in the faeces
Bl
on the 11th day (Jungherr, 1948); while
virus
adults ceased to spread virus after
by contact
(Walker and McKercher,
34 days
1952) multiplied first in the lungs, and nine to ten days later was eliminated in
1954b). Experimental
the faeces.
reviewed above suggest that the permanent carrier state in Newcastle disease is rare
Complete fied
live
ministered
absence of spread of a modi
Newcastle intranasally
from the vaccinates
disease
vaccine
ad
in
chickens
and field data of the type
(Brandly,
1950;
or intramuscularly
1952;
to
Konev, 1953; Levine, 1954; Placidi and Santucci,
non-vaccinated
chickens within the same pen or separated
Hofstad,
1951;
Crowther,
Jungherr, 1948; 1952; Nitzschke, 1955; Piatt,
32
J
1948). Even if this
is true
for chickens,
there may be a more lasting carrier
state
in turkeys.
A
did not spread to susceptible chickens and
which did not cause parental antibodies in young
for the patho genesis of Newcastle disease virus follow scheme
suggested
and alimentary
ing respiratory
has been given by Kohn
infection
believed
virus
in the intestinal
after respiratory
infection
upper respiratory
occur (Figure 6). The
tract would
tract has been consider
has also been reported
methods.
The possibility that hens can carry non
(1955,
1959) that multiplication of the
who
chicks
(Bankowski, 1961c). This unusual type of infection was identified by serological
Newcastle
pathogenic
excrete this virus discussed
disease virus
in their eggs
(1957a). Similar
by Sinkovics
latent Newcastle
and
has been infections
disease virus
ed the site of election for the early multi
have been found in cultures of tissue cells
plication of the virus Coffin, 1961). Kohler
inoculated
that Newcastle
disease virus was ingested
with a strain of virulent virus (Mason and Kaufman, 1961a). Further mention of asymptomatic infec
by leucocytes,
but that the virus did not
tions is made on page 57.
(Maestrone and (1960) reported
appear to multiply within these leucocytes.
Transport of Live Poultry Reservoir of Virus in Aqueous
The movement of domestic poultry has
Humour
been a very important
Clark et al. (1955) found that the aqueous humour of the chicken eye was a source of virus for 10 days after intra muscular inoculation.
They reported later (1957) that they believed the aqueous humour was a reservoir for Newcastle disease virus in a large percentage of in California. However, chickens more recent
findings
material
have
indicated
does not constitute
that this
a permanent
reservoir of the virus (Baldwin, Dardiri et al, 1959; Fritzsche, Pannu and Bankowski, 1962).
1962; 1961;
infections
with
New
castle disease virus have been reported in
which spread of virus could be detected. For example, chickens hatched two or three months after an outbreak of clinical Newcastle disease had subsided proved to be serologically positive. Although no clinical evidence of disease had been observed
in the young stock, it was con
cluded that inapparent
has been reviewed 1949a,
1950,
(1943,
by Beaudette
1951b),
Brandly
et
al.
(1946a) and Brandly ( 1950, 1959). More recent reports include those of Anon.
(1961a), Crowther (1952), Scott et al. Reid (1961), Vandemaele ( 196 1 ) , and Winmill and Haig (1961). Of 2,489 outbreaks of Newcastle dis ease in Britain, 56 per cent were a result of traffic in live poultry. Field evidence ( 1956 ) ,
in these
suggested that recovered
outbreaks
some
(Asplin
birds acted as carriers
etal,
Asymptomatic Infections Asymptomatic
factor in the spread
of Newcastle disease. Spread by this means
spread by carriers
had occurred (Jungherr and Terrell, 1946). Asymptomatic Newcastle disease which
1949). Levine (1952) considered
infected source
bird
the actively
to be the most
of infection.
( 1941 ) , in discussing the original at
Ranikhet,
infection
outbreak
India, suggested that
the
been
the
might
importation
important
Similarly Haddow
have
of poultry
due
to
from abroad four
months previously.
Poultry Markets The significance
of poultry markets in
the spread of Newcastle recorded
disease has been
in all geographical
regions since
the disease was first recognized
(Doyle, 33
1927; Albiston and Gorrie, 1942; Farinas, 1930; Lissot and Moessel,
1949; Seethara-
1951b). In Australia, almost every outbreak has been associated with markets man,
(Johnstone, 1931). An analysis of 602 outbreaks in England and Wales showed 158 to be primary cases; and from these primary outbreaks a further 444 outbreaks resulted — mainly from local spread, direct dealers'
sales,
transactions
through markets (Reid,
of Newcastle
breaks
and
1 955).
sales
Other out
disease have
been
Illegal Movement Finally, the illegal movement of live birds from infected premises or areas has resulted in new foci of infection (Jansen and Kunst, 1952; Reid, 1955, 1961).
Poultry Carcasses and Offal Poultry carcasses and offal have been great a source of Newcastle
as
disease infec
tion as live poultry and they have often carried the disease from one country to
traced to a poultry packing station (Keymer, 1961) and to the purchase of fowls
another. In Switzerland,
from ships in dock (Lucam, 1949b).
chickens
Laying Trials
fection, a diagnostic
method was develop
ed to detect infected
imports of chicken at
Newcastle
disease outbreaks
with the movement
associated
of live poultry have
been
where offal from
of foreign origin has been con sidered a highly important source of in
the
1963). From 30 frozen consignment,
frontier (Hess,
imported
per 5 tons of product
every heads
reported at laying trials (Beach, 1949a, b; Piatt, 1948). In some trials, birds were identified which had recovered from
the
Newcastle
disease but which were, pre sumably, not excreting virus (Asplin et al., 1949; Reid, 1955; Jungherr and Terrell,
into young susceptible chicks. During the period 1947-62, samples from 21,300 tons of imported chicken were examined in
1946). When these birds were traced, a subacute form of Newcastle disease was
Switzerland
in a hatchery (Anon., 1962b). Furthermore, the tracing of birds moved from a large poultry show in Britain dis closed 255 outbreaks of Newcastle disease discovered
(Reid,
1955).
laboratory.
For
were examined in the examination,
material from the spinal cord was injected
(Hess, 1963). Similarly in Austria, the importation of refrigerated poultry carcasses has been considered the most important source of initial infection (Grausgruber, 1963). In Germany, the laboratory examination of imported poul try carcasses has yielded a strain of New-
4 — Probable Origin of the First 542 Outbreaks of Newcastle Disease occurring in England and Wales during 1947
TABLE
(Gordon et al., 1948)
Traffic in live poultry Feeding infected material Local spread Mechanical spread Obscure 34
Origin
Per cent
sales, auction markets and pet shops Unboiled swill, poulterers waste and table poultry trimmings Contiguous premises and mixing and/or straying of poultry Visits and handling of birds by dealers and others, infective clothing, crates, etc. —
42
Dealers
33 28 7 10
TABLE
5
— Newcastle Disease Virus Isolated from Poultry Carcasses Imported into England in 1949 (Dobson, 1952) Per cent carcasses infected
Young fowls Hens Old cocks
69 66
80
Geese
6.9 11
Ducks Turkeys
24
for chicks (Hartwigk and Gothe, 1958). Poultry offal has been an important mode of spread not only between countries but also within a country. Doyle (1927) mentioned that the flock involved in the original 1926 outbreak in England had been fed offal collected from the seaport town of Newcastle. The origin of the 1933 outbreak in England was not determined (Dobson, 1939). However, in the 1947 outbreaks in England there was evidence that poultry had had access to uncooked offal from uneviscerated poultry carcasses imported 13 days previously from Hun gary, Poland and other countries where Newcastle disease was known to exist (Andrews, 1948; Gordon and Asplin, 1947; Gordon et al., 1948; Reid, 1961). In 33 per cent of the first 542 outbreaks, there was a history of poultry having had access to offal from imported carcasses (Table 4). Less than 5 per cent of later
castle disease virus pathogenic
resistant fowls; of processing in an infect or of surface contamina
ed environment; tion
from
a smaller
number
of infected
fowls.
Topacio and Coronel (1939) found that the crop contents and the organs of dead birds
remained
infective
for seven days
after death. In frozen carcasses, the virus has been recovered from the brain (Hess, 1963; Tienfenbacher and Woernle, 1957) from spinal cord material (Hess,
and
1963). The spread of Newcastle disease virus from stored edible carcasses to a pen
of pullets has been described by Gordon and Asplin (1947). Frozen poultry carcasses are also to
thought
disease
to
have
introduced
Germany
Newcastle 1942;
(Koser,
1948), Sweden (Alegren, 1951; Bakos and Nordberg, 1949; Isaksson et al., Wagener,
(Hess, 1951). 1948) and Switzerland Samples taken from chicken carcasses im resulted in the ported into Switzerland disease virus from
of Newcastle
outbreaks were associated with the feeding
isolation
of poultry offal, whereas outbreaks origi nating from stock movement increased to
350 consignments
over 70 per cent (Gordon et al., 1948). In a 1949 survey of samples of skin
(1963) believed that the absence of New castle disease in Switzerland was largely
taken from frozen carcasses imported into England from two countries, Newcastle
the result of testing samples of imported
disease virus was recovered
from several species of poultry, as shown in Table 5 (Dobson, 1952; Dobson and Simmins, 1951; Reid, 1961). Asplin (1952) con sidered result
that these findings
of
subclinical
infection
could
be the
in partially
representing a total of 1,900 tons of product (Hess, 1963). Hess
poultry carcasses.
Under virus
experimental
has remained
conditions,
viable
in
the
the bone
marrow of fowls for 300 days or longer, on the storage temperature (Table 6). These findings explain how the holding of live poultry at killing plants depending
35
TABLE 6 — Reports of Duration of Viability of Newcastle Disease Virus in a Variety of Tissues Author
Storage
Material
Duration viability
temperature
(days)
Asplin, 1 949
Beach, 1943a Dalling, 1960 Doyle, 1 933 Doyle, 1 927 Iyer, 1 940 Hess, 1951 Hartwigk and Gothe, 1958
Hartwigk and Gothe, 1958
prior to slaughter (Reid,
Bone marrow Bone marrow and skin Skin Lungs Poultry carcasses Bone marrow and muscle Kidney of infected fowl Liver and spleen of infected fowl Poultry carcasses Infected poultry carcasses Infected poultry carcasses
has facilitated
spread
reports
on
34-35"F — 4°F
Frozen
134-196 Over 300 98-1 60 60-90 Over 730
Chilled Approx. 2°C 17'C
Over 1 00 21-28
34-35'F Live chickens
180
180 Over 836
Frozen
— 20°C Buried in earth
the
121
of Newcastle
isolation
disease virus from a case of conjunctivitis
1961).
Although, in general, studies of virus
Foster and Thompson ( 1957) indicate that the less virulent strains would probably
man was written by Ingalls and Mahoney (1949). Steele (1959) has pointed out that in respect of Newcastle disease infections, the milder the strain of virus the broader the host spectrum. Thus, the velogenic strains
survive just as long.
seldom affect man, whereas the lentogenic
survival with
in carcasses have been concerned
the more
virulent strains of New
castle disease virus,
results
reported
by
in
Bl
Spread by Human Agency Outbreaks chicken
of
Newcastle
disease
in
flocks have been associated with
from infected premises (Lancaster, unpublished data). The move ment of seasonal land workers from Italy was probably responsible for introducing the disease to Germany (Wagener, 1948). Also, Hudson (1937a) and Callender
human movement
(1958) have reported that infection can easily be conveyed from sick to healthy stock by handling.
influenza
virus
from human
cases has shown these to be neutralized
by
Newcastle disease serum and to have other resembling Newcastle virus (Solov'ev etal., 1963). properties
The disease in man may like
disease
be an influenza
accompanied by headache (Howitt et al, 1948; Negri et el, 1953). On the other hand, Newcastle disease virus has
illness
been isolated illness
from
was
a human
almost
patient
inapparent
(Quinn et al, 1952). Although they did not isolate the virus, Howitt et al. (1948)
Reviews of reported cases of Newcastle disease in man have been published
by
(1955a), Hanson
and Brandly (1958), Sinkovics (1957a) and Thompson ( 1 950) . One of the earliest }6
of atypical
whose
Human Infection
Blood (1950), Evans
strain has caused severe conjunctivitis
and malaise in man (Dardiri et al, 1962). Furthermore, re-examination of 1 7 strains
have
suggested
that
Newcastle
disease
virus may be responsible for some atypical central nervous system infections in man.
The more common symptom
is a unilateral
conjunctivitis
and
(Gustafson
Moses,
1951; stance,
Jacotot et al, 1950). In one in conjunctivitis due to Newcastle
disease
virus
developed
in
laboratory
a
at a dilution of 1 : 10 or (Collier, 1951). In assessing the significance of such findings, it must be
haemagglutination higher
worker who had been infected in similar circumstances four and a half years earlier (Jacotot et al, 1955). Other cases of con junctivitis, with or without enlargement of the adjacent lymph nodes, have been reported by Anderson (1946), Burnet (1943), Freymann and Bang (1949), Ku(1952), (1948), jumgiev Lippman Reagan et el. (1956), Schoop (1954) and Slonim and Stranakova (1952). In many cases of human infection, an
emphasized that there is evidence indicat
immune response has been observed (Bang and Foard, 1956b; Blood, 1950; Hunter
due to common
1951; Jordan
et al.,
and Feller,
1950). Also, inclusion bodies have been seen in
of epithelial cells from the
the cytoplasm
though their significance
conjunctiva,
has
not been fully established (Hunter et al., 1951; Keeny and Hunter, 1 950; Orlandella,
1955). In some instances, human infec has followed the accidental inges
tion tion
of
Newcastle
disease virus (Divo and
Lugo, 1952). During the acute phase of infection, the virus has been recovered from eye wash ings (Jacotot et al., 1950), blood (Negri et
al,
1953; Reagan
et
al,
1956),
urine
that
ing
virus
mumps
and
antigens. Thus, neutralizing castle
disease
virus
were
present in half a group
from mumps
valescing
and Newcastle
disease viruses cannot antigens (Wenner et
1952a, b). A heat stable factor, with complementfixing
as
activity
well
antihaemagglutinating disease virus,
as
Newcastle
against
increased in titre in the sera of a number
of
from mumps patients convalescing (Jordon and Feller, 1950). Similarly, sera from mumps patients have shown New
castle disease virus antibody by
virus
when tested
(Kilham et al, a viricidal factor for
neutralization
1949). In addition, Newcastle disease virus and a component
haemagglutination by that virus have been identified in the normal sera of inhibiting
(Howitt,
man and some other mammals it has
al,
1952a, b). Therefore, been concluded that the serologi
1950; Wenner et
disease in man
must be made with caution in the absence
in the evisceration
of poultry
of virus isolation (Evans, 1956; Jordan and Feller, 1950; Jungherr et al, 1949; Kilham era/., 1949). It has been emphasized that in many
et
survey
al,
of 1,363 samples of human
revealed
13
to
contain
haemag-
glutination-inhibiting and complementfixing antibodies against Newcastle disease 1957). A similar survey that 50 per cent of human sera
(Scatozza,
showed
be
al,
40 persons
rier state in man. Neither is there any evi dence of transfer from one person to an (Yatom, 1946), other this although possibility has been discussed (Hanson and Brandly, 1958; Mitchell, 1953).
virus
to be
involved
1952). Nevertheless, there is no evidence of the existence of the car
serum
New
shown
1949). In contrast, other reports have indicated that cross reactions with mumps
disease conjunctivitis
A
against
of patients con (Jungherr et al,
cal diagnosis of Newcastle
(Nelson
and antihae-
antibodies
magglutinating
(Dardiri et al, 1962), saliva (Divo and Lugo, 1952) and respiratory tissues. In one instance, an outbreak of Newcastle employed
Newcastle antigen or
disease virus have a common
samples inhibited
Newcastle
disease virus
human
infections
with Newcastle
disease
virus, the antibody response has been low (Anderson, 1946; Freymann and Bang 1949;
Slonim
and
Further, Newcastle
Stranakova, disease
virus
1952). may
share antigenic components with other in fectious agents (Evans, 1951). Thus, it has been shown that serum from human patients blood
from infectious will agglutinate human red
convalescing
mononucleosis cells
modified
with
Newcastle 37
(Florman,
disease
virus
capacity
to agglutinate
virus-treated
cells is unrelated to the anti
body of infectious and Curnen, ing property
This 1949). Newcastle disease
mononucleosis
from the virus
distinct
(Evans
1948). The red cell sensitiz of Newcastle disease virus is haemagglutinin
(Evans, 1955b).
have
different
been
(Verge, indicate
used
species of mammals in
studies
laboratory
1948). The results of these studies that the ability of Newcastle
disease virus
to multiply in mammalian
hosts is limited
(Hanson
et
al,
1951a;
1952b). In general, New castle disease does not appear to be spon Reagan
et al.,
taneously transmissible
to mammals under
1948). Up to now, man has been the only mammalian natural
conditions
(Verge,
species in which a number Newcastle
of
disease virus infection
cases
and Guinea- Pigs
Although ferrets have been found ceptible
Newcastle
to
disease
of
have oc
Also, curred naturally (Blood, 1950). there is no evidence as yet to indicate that mammals play a part in the spread of the disease. However, according
Reagan et al. (1950c) have reported that the virus could not be adapted to this species. In some transmission have proved tible (Wenner and Lash, guinea-pigs
experiments,
to be insuscep 1949).
A
number of workers have reported the
of hamsters
susceptibility
Newcastle
to
disease virus. Thus, Reagan et al. ( 1 947b, 1948a, 1950c) passaged a virulent strain
of virus for 300 passages in the brains of Syrian hamsters. The virus produced and symptoms of irritability, paralysis rapid death (Reagan et al., 1947a). The amount of virus required to produce in fection in hamsters varied with the route inoculation of inoculation. Intracerebral was found a successful route (Prudovsky et al., 1961), and 10"10 ml. of virulent virus killed hamsters within 4 to 16 days when administered
by this route (Mitroiu
and Vior, 1960). Following the intracere bral inoculation of attenuated Newcastle
to Brandly (1959), the studies described below in dicate the potential range of infectivity of
challenge infection was produced
the virus. Other references to the propaga
strated by virus neutralization
tion of Newcastle
sus
virus,
Hamsters
Warm-Blooded Animals Several
Ferrets
disease virus in mam
malian tissue are given on page 1 18.
disease virus, an active immunity against and Vior,
1960);
(Mitroiu
this was also demon tests
(Rea
gan etal., 1947a).
Horses Bats
Horses have been inoculated
The cave bat (Myotus lucifugus) has been found susceptible to intracerebral inoculations with a number of different strains of Newcastle disease virus (Reagan et al., 1950a). The virus has also main tained its pathogenicity for chicken em bryos following intracerebral passage in bats (Reagan et el., 1951a). The same species of bat was susceptible to the Cali fornia strain 11914 of virus administered by intranasal instillation,
and typical symp
toms of irritability, paralysis and death re sulted (Reagan
and Brueckner,
1951a).
castle disease virus
(Lulic,
mune serum is discussed
further
to produce
with New hyperim
1955). This subject on pages 87 and 88.
Mice The successful adaptation of the virus mice following intracerebral inoculation has been reported (Kilham, 1950; Sinkovics, 1960). This mouse-adap
to unweaned
ted
virus
did not propagate
in the brain
of adult mice and there was evidence of decreased pathogenicity for chicks (Sin kovics, 1960).
Monkeys The intracerebral
of Rhesus
inoculation
monkeys with Newcastle
disease virus has
resulted
in a meningoencepha litis (Prudovsky et al., 1961; Rodot, 1953; generally
Wenner and Lash, 1949). Intramuscular injection has produced little or no clinical effect (Reagan et al., 1951b). Sublethal infections in monkeys have resulted in the appearance of haemagglutination-inhibiting antibodies
(Wenner
et al.,
ported
a strain
1952a).
(Reagan (Marina brevicauda) and Brueckner, 1951b) and rabbits (Wil cox et al., 1958) have also been used in laboratory studies.
The susceptibility
of the domestic cat
to Newcastle disease virus
has been re
by Feldberg and Luttrell (1958); and Bolin (1948) isolated Newcastle disease virus from the faeces of a cat ported
months
chicks.
after
was
it
Furthermore,
fed
infected
Nakamura
and
Iwasa (1942) believed that a laboratory cat died of a natural
Newcastle experiments
infection
caused by
disease virus. Subsequent on the infection of cats per os
showed
that these animals are susceptible to the virus: Luttrell and Bang (1958)
and Tokuda (1956) reported that young and adult cats could be infected with New castle disease virus administered by differ ent routes, and Verge (1948) also con cluded that cats were naturally susceptible to the virus. However, these results were not confirmed by Reagan et al. (1954d)
who found that kittens were not suscep tible to infection with a strain of virus ad ministered by the oral or nasal routes, but were
susceptible
oculation (Reagan
to intracerebral
in
et al., 1954a).
Dogs and Foxes Newcastle disease virus has been re from the urine and faeces of a
covered
the
of dogs and cats 1954a; Mitev et al, 1958;
insusceptibility
(Bonaduce,
1954; Rahneberg,
1960; Rea
1952).
Hedgehogs The finding, in Morocco, that a North African hedgehog (Aethechinus algirus) became infected
after eating another that
had died from the disease has suggested that this animal
Cats
re
intracerebral inoculation of Newcastle disease virus. In
gan and Brueckner,
Shrews
Sil-
were
puppies
contrast, other reports have indicated relative
day that
to
susceptible
with
that young
finding
Orlandella,
Shrews and Rabbits
five
fox and a dog on the first and fifth after feeding on carcasses of fowls died of Newcastle disease (Polci and vagni, 1954). Baldelli (1955) has
might act as a reservoir
of the virus in Morocco and also, perhaps, other countries (Placidi, 1954a). Pigs In studying domestic
the
part
played
by the
pig in the spread of Newcastle
disease, Asplin (1949) found that, follow inoculation, virus was ing intramuscular
of one pig for a of 48 hours. The experimental inoculation of Newcastle disease virus into excreted
in the urine
maximum
young pigs has resulted in death 3 to 11 days
later
1954b), symptoms
(Buck virus
(Hofstad,
Placidi, of nervous
1950;
or the development
(Bindrich, 1957) and paralysis
et al.,
passaged
1954a). Newcastle disease in young intracerebrally
pigs for eight passages has resulted in the virus becoming
pathogenic
for the pig via
the nasal route. Fourth to seventh passage virus has produced neither disease nor im munity in chickens (Buck et al., 1954b). The absence of Newcastle disease haemagglutination-inhibition antibodies in 168 pigs would suggest lack of contact with the virus. The subcutaneous injection of an avirulent
Newcastle
disease virus
into
a
young pig resulted in the development of a positive but low Newcastle
disease
HI
titre
(Dyml, 1958). 39
Rats
injection
The oral dosing or feeding of the com mon brown rat with Newcastle disease virus or with chickens dead from the
goldfish
disease has resulted in the virus being shed in the excreta for 24 hours (Asplin, 1949), 48 hours (Zuydam, 1951a), 72
hours (Walker et al, 1954c) and 5 days (Baczynski, Newcastle 1959). disease virus was not isolated from faeces and tissues of 12 rats taken from a Newcastle disease-infected farm (Zuydam, 1951a). From these studies it was concluded that, although
the rat is generally only a me chanical carrier, it could be responsible for causing the disease in chickens
(Placidi andSantucci, 1953c). Cattle Two cases of naturally occurring infec tions with Newcastle disease virus have been reported in calves (Ozawa and Chow, 1958; Yates et al, 1952). In both instances the virus was isolated from the lungs of dying calves which had respiratory symp toms. Although calves have been artifi
cially infected with Newcastle (Helmboldt et al, 1955),
virus
disease there is
no confirming evidence that cattle play any part in the spread of the disease. Several studies have been made on the propagation of Newcastle disease virus in the lactating mammary gland of cows and goats (Mitchell et al, 1953a, 1953b, 1954,
1956,
1958;
Easterday has persisted
et
al,
1959). The virus in the mammary gland for varying periods up to approximately two weeks and, in most cases, it has stimulated the production of antibody. The mammary has appeared to be the principal
neutralizing gland
site of antibody
al,
production
(Mitchell
et
1956).
Cold- Blooded Animals Reptiles and Fish (1957) found no evidence of a carrier state following the intracerebral Qureshi
40
of Newcastle (Carassius
disease virus
auratus),
into
garden
(Thamnophis sirtalis) or baby (Graptemya geographicalesuerr) . However, Placidi (1956a) has reported that the virus of Newcastle disease could be recovered for several months following the intracerebral inoculation of the tor toise (Testudo graeca and Clemmys leprosa) and three species of snakes {Natrix natrix, Natrix viperina and Malpolon monspessulana) . The virus has also been snakes
turtles
propagated in the green turtle (Pseudemys elegans) (Reagan et al, 1953).
The intracerebral inoculation of New castle disease virus into the dog fish (Scyllium canicula) has been followed by re covery of the virus during the subsequent three weeks. Lesions of meningitis have been found
in the brain, but without
any
clinical symptom (Atanasiu and Atanasiu, 1955).
Invertebrates Insects. on
Studies
ectoparasites
on in
invertebrates, particular,
and
indicate
that they play a similar role to mammals in the spread of Newcastle disease. Hofstad (1949a) showed that the northern feather mite (Liponyssus sylviarutn) har boured the virus after feeding on infected birds during the viraemic stage. However, the bite of known infected mites failed to transmit
Newcastle
A
disease to suscep
similar absence of natural transmission has been reported for the tick (Argas persicus) (Komarov, 1940; Seetharaman, Lissot and Moessel 1951b). (1949) were of the opinion that ectopara sites (genera Menopon and Lipeurus) played no role in the spread of Newcastle disease. However, Bolin (1948) has re ported the isolation of Newcastle disease virus from common chicken lice collected from hens 35 days after infection with the virus. In the early outbreaks in the Dutch tible birds.
East Indies, flies and mosquitoes apparent ly played no part in spreading the disease
(Picard, 1934).
Worms. In earthworms (genus Helodrilus), Newcastle disease virus has been found to survive approximately four days (Baczynski 1960b; Boyd and Hanson, 1958), or 18 days if the worms were kept at 21°C (Boyd and Hanson, 1958). Al virus
though
fed
to planaria
(Planaria
ers or pens two to seven weeks after the
of infected
removal
(Dobson,
chickens
1939; Jungherr, 1948; Jungherr and Ter rell, 1946). Levine et al. (1950) con cluded that premises with birds recovered
from Newcastle
disease do not harbour
virus to infect susceptible
sufficient
birds
after the flock recovers from
maculata) has been recovered after nine days there has been no evidence of virus multiplication (Boyd and Hanson, 1958). The part played by the large intestinal round worm (Ascaridia galli) in the transmission of Newcastle disease virus
one month
was examined
during winter (Fortner, 1952). Schyns and Florent (1951) and Hudson (1937a) also found the virus in uncleaned pens to be inactive after 6 to 14 days. In contrast,
by Stefanski and Zebrowski
(1958) who found that the virus could be recovered from these nematodes re moved from chickens dying from New castle disease. The virus was not recovered from the eggs of the worms, and there fore A. galli was not considered a true vector
of
Newcastle
Snails. disease
The
virus
remained
that
Newcastle
alive for several
weeks in the body of the Indonesian
giant
snail (Achatina fulica) has suggested that the snail might spread the virus (Mansjoer, 1961).
Inanimate Causes Infective secretions and
respiratory
tracts,
and,
of
to a lesser
from the reproductive tract of poultry, are the chief causes of contamina tion of buildings and equipment. Various extent,
chemical
and physical conditions
influence
the viability of the virus outside the host.
Therefore, much of the data referred to below relates only to the particular
experi
The survival of Newcastle disease virus in the
mental
and field conditions
environment its
involved.
plays an important
perpetuation
and
spread
part
in
(Hanson
and Brandly, 1958).
Chicken Houses, Crates and Brooders Day-old chicks have become infected when placed in uncleaned
symptoms.
In
poultry
occupied
by
infected
previously
stock, Newcastle disease virus has sur vived no more than 7 days during sum mer, 14 days during spring and 30 days
Michalov and Vrtiak (1963) found that virulent
disease virus survived
Newcastle
53 days in hen house litter.
Using strains et al.
(1957)
failed to recover the virus after 24 hours from sterilized dried droppings, shavings, feathers
and
feed
samples
been placed in a brooder
which had
house. Hitchner
(1950) also reported failure of the Bl strain of vaccine virus to survive in a hen house after removal of the vaccinated chickens.
and excretions
disease virus from the digestive
Newcastle
respiratory
houses
of vaccine viruses, Dardiri
disease.
finding
the
battery brood
Under experimental
conditions,
New
castle disease virus on small amounts of in stoppered vials has re chick down mained viable in a hen house for 255 days (Olesiuk, 1951). At 37°C, Newcastle disease virus
has
remained
viable
in the
of an egg after 94 to 126 days (Olesiuk, 1951) or on pieces of egg shell for 1 day (Asplin, 1949) or 7 days (Olesiuk, 1951). The virus has remained viable on non-sterile burlap for 20 days
contents
(Jungherr, 1948); and on sterile burlap from 55 days (Jungherr, 1948) to 108 days or longer (Olesiuk, 1951), depend ing on the storage temperature. It has been shown
that Newcastle
disease virus
will survive for at least 24 hours on pieces of egg shell when exposed to incubator conditions (Asplin, 1949; Beamer et al., 41
TABLE 7 — Duration of Viability of Newcastle Disease Virus (Modified from Olesiuk, 1951) Duration of study: 538 days Incubator
Test material
37°C
(days) Burlap
12
Filter paper
19
Normal room Hen house 20° to 30"C — 11'to 36'C (days) (days)
Refrigerator 3° to 6'C (days)
Deep freeze — 26X
123 157
538 538 538 538
108
13 37
129
Cloth
13
44
145
193
Broth
30 26 7 54
152
538
44
228 228 228
61 83
172 172
58 14 110
163
255
66
172
216
255
258 258 538 538 235 538
33
80
199
258
Saline Eggshells Mash Chicken Faeces Down Soil Eggs A.A.F'
79
41
451
(days)
538 538 538 538 538 538
—
538
1 Undiluted amnioallantoic fluid
1949). The duration of viability on other is given in Table 7.
test materials
Asplin (1961) considered that survival times
obtained
probably
longer
in
the
were
laboratory
than would be expected
from field experience. Under farm condi tions, the of decomposing presence organic variations
matter,
exposure
to
light,
in humidity probably
the survival
time.
Nevertheless,
and
of spread of the infection to a second farm on which 72 per cent of 18,000 chickens subsequently died (Jungherr and Markham, 1962). Farinas (1930) was able to infect chickens, geese, turkeys and ducks by using boxes that had previously contained
native
chickens
in
bought
a
public market.
shorten
Olesiuk
(1951) considered it likely that the virus would survive in the natural environment from one season to the next. The survival of Newcastle disease virus in the soil depends on several factors, but has been found to be from 8 to 22 days
(Boyd and Hanson, 1958). Survival and spread outside the host is also facilitated
Wind In
areas where the poultry population
disease virus may be from one location to another by the wind (Anon, 1962b; Callender, 1958; Moynihan et al., 1951). This wind-borne spread has been particularly noticeable is dense, Newcastle
carried
in
countries
where
large
numbers
of
by the high resistance of the virus to direct
chickens are housed in buildings ventilated
sunlight (Iyer, 1943; Skinner and Bradish,
by
1954) ; to a wide range of pH values, from pH 2 to 11 (Moses et al., 1947; and to
between
effects of temperature
1946;
Temperature
is
(Brandly, 1959).
an important
factor
and
in Table 8. Empty poultry crates returned from a farm on which an outbreak of Newcastle disease had occurred were the likely means its effect is summarized
42
exhaust
(Reid,
fans
(Jungherr and Terrell,
been recognized 1961
1961). Spread premises has also
contiguous
Moynihan
et
al,
1951;
Reid,
), although the exact means involved
was not ascertained. data supporting the field Experimental evidence have been obtained. DeLay et al.
(1948), DeOme
et al.
(1948) and Mans-
8 —
TABLE
Effect of Temperature on Infectivity of Newcastle Disease Virus (Modified from Anon., 1959)
Temperature Probable stability in time
60
140
56
132
45
113
37
98
0-5 minutes 30-180 minutes
Hanson Hanson
12-90 hours
6-14
Authority1
days
— Roakin strain, 5 min. (A) — GB strain, 180 min. (A)
Roakin strain, 30 min. (A) B1 strain, 1 5 min. (A) GB, Roakin and B1 strain, persisted 1 2 hours but not 96 hours. (A) GB, Roakin and B1 strains, persisted 14 days. (A) Conn, strain, 6, 24 and 72 hours. (C) Mass. strain, 10 days. (C)
Hanson
—
Hanson
—
Jungherr Olesiuk Torlone
— — — Strain
30-1 00 days
77
Prier
Olesiuk Torlone
24
strain, 73-95 days. (A) — Mass. strain, 9-16 days. (C) — Strain F — titre reduced after
48
— NDV
hours.
0
9 months
32
— 20
1-10
—4
F
titre
reduced
(A)
(A)
Lancaster — Strain F, 30 days. (B) Jungherr — Conn, strain, 203 days. (C) (on filter paper 34F) — NDV strain, 80-1 1 9 days. (A) Prier — Mass. strain, 123 days. (C) Olesiuk — B1 strain, titre reduced in 6 months. Reising Lancaster — Strain F. (D)
8-1 6 months
41
—
after
hours.
25
and record
Hanson Reising
years
^
— NDV
strains, 4-6 years. (B) — B1 strain, high titre retained 1 year.
(B) Lancaster — Strain F, 1 year. (D) A — Allantoic B — Allantoic C — Allantoic D — Allantoic
fluid fluid fluid fluid
in sealed glass vials. in stopper glass tubes. dried on paper or cloth.
lyophilized.
1 Hanson, 1 955 ; Jungherr, Reising
joer
1 948 ; Lancaster, and Hitchner, 1954; Torlone, 1955.
(1961)
of Newcastle samples
demonstrated
from
collected
a
poultry house and Idnani
diseased chickens;
containing
and Seetharaman chickens
the presence
disease virus in air and dust
kept
(1947) used susceptible distance from an
a short
infected bird.
Aerosols Artificially-
aerosols
and
transmit
naturally-produced the
disease
readily
(Andrewes and Allison, 1961; Hanson and
1 957a ; Olesiuk, 1 951 ; Prier and Alberts, 1 950 ;
Sinha, et al.
1952; Sinha (1952,
1954)
al, 1957). Sinha recovered Newcastle
et
disease virus from the air in the vicinity of infected chickens on the third to sixth day after exposure. Under experimental conditions, it was shown that with each minute of aerosol exposure, chickens would be expected to inhale five particles of Newcastle disease virus (Sinha et al., 1954) . Such low concentrations of virus in the air may be readily
etal,
infective
(Nadel
1957). 41
Poultry Vaccines as a Means of Spread of Newcastle
Water The spread of Newcastle water
contaminated
disease
introduced
by
irrigation of meadows was considered Koser (1942). Similarly, likely by Grausgruber
(1963)
rivers contaminated
considered
that
by infected carcasses
represented a means of dissemination. Kraneveld and Mansjoer (1950b) found that water contaminated with faeces from infected birds remained infective for at least five days. Idnani and Seetharaman
(1947) also showed that chickens could be infected by drinking contaminated water. In the Philippines, an important source of infection was thought to be the common drinking ponds (Farinas, 1930). Thus, Kee (1928) found that healthy chickens
confined
in separate pens near
the diseased birds, but with separate drink ing troughs,
did not contract
Newcastle
It has been shown experimentally disease virus will survive
Newcastle
19 days in lake water, depending
pH,
that 1 1 to
on aera
presence of salts and organic matter (Boyd and Hanson, 1958) and undetermined factors (Winterfield and Seadale, 1956a). In contrast, it has the
been possible to recover Newcastle
disease
vaccine virus placed in drinking water in a brooder
Failure to inactivate
a virulent
castle disease virus completely
paring a killed vaccine has led to some serious results in the field (Mitchell et al., 1952; Placidi, 1956b; Surin, 1959). Henderson
(1952) considered that
the
absence of active virus in an inactivated vaccine could never be proved absolutely.
Furthermore,
instances
where commercially
occurred
have
manufactured
New
castle disease, fowl pox and laryngotracheitis
vaccines
contaminated disease
virus
1950a;
a virulent
Newcastle
and
Pomeroy, 1959; Hofstad, 1954a;
(Zargar
Brandly,
Marek,
have been found to be
with
1961). Furthermore, a poultry with Newcastle contaminated
vaccine
disease virus
is
known to have resulted
house for only 24 to 36 hours
free area (Anon., 1964). The difficulty of detecting a pathogenic Newcastle disease virus in a live wing-web
previously
Newcastle
disease vaccine
has been em
(Rosenwald et al., 1959). Also, certain strains of mesogenic vaccine virus phasized
(for example, MK107) inoculated cages
clinical
have spread from
birds to contact birds in other
of the same unit.
was
no
et al., 1959). Under the route of spread was
(Dardiri et al., 1957; Luginbuhl et al., Marek, 1960; Winterfield and
these conditions,
Seadale, 1956a).
considered to be aerogenic.
sulted
There
effect, but an active immunity re
1955;
44
New
when pre
in the introduction of the disease to a
disease.
tion,
Disease
by the
(Rosenwald
II:
PART
Diagnosis
GENERAL CHARACTERISTICS The was
causal agent of Newcastle
first
identified
by
Doyle
disease
(1927)
who showed that the virus was distinct from the virus that caused fowl plague. This finding was based on six main factors: period of incubation, symp lesions,
post mortem
toms,
blood
infec-
OF THE DISEASE
lesions were in the (Capobianco, 1949). In The Netherlands, the initial severe form of the disease has been followed by an in crease in the number of mild or subclinical outbreaks (Hoekstra, 1961). Similarly in western Germany, the character of New the main pathological tract
alimentary
tivity, disease infectivity and results of cross-immunity tests. Later, following an of virus strains between interchange workers in Java, the Philippines, India,
castle disease has changed and the appear
Japan and England,
In contrast, the epizootic of Newcastle disease which occurred in Kenya in 1955 was equally as virulent as the epizootic which occurred 20 years previously (Scott era/., 1956). The disease now considered to have four main forms: the velogenic (virulent), the mesogenic (less virulent), the
showed
tests
six virus strains (designated Java, Philippine, Ranikhet, Japan, Korean and Newcastle) were immunologically indis tinguishable (Doyle, 1935). The marked difference between the clinical features of the classical or original
that
(
Beach,
Newcastle
disease and the disease
"avian
pneumoencephalitis"
943
delayed for seven years the
)
of
1
form
termed
of the latter
picture (Fritzsche,
and pathological
1963).
is
neutralization
and
lentogenic
and
(mild)
matic. This classification, is
serum-virus
cross-immunity
ance of lentogenic and mesogenic strains of the virus has complicated the clinical
useful for descriptive
the
asympto
though arbitrary, purposes and for (Lancaster,
grouping
breaks
domestic poultry such as ducks, geese and turkeys which are more resistant to the
or outbreaks with no characteris
tic symptom or lesion (Beach, also had an important nosis and diagnostic
bearing
1946), has on diag
methods. a
Another feature that has affected diag nosis has been the recognition of chang ing clinical picture (Brandly, 1953). In Yugoslavia, for example, the incidence of the nervous form of the disease has in creased (Jaksic and Stefanovic, 1957). In Italy, initial outbreaks generally involved the upper respiratory tract; whereas later
is
It
virus.
Incubation
Period
After artificial infection, the incubation from to 25 days has varied (Cooper, 1930; Doyle, 1927; Farinas, 1930; Guha and Chatterjee, 1950; Jungherr et. al, 1946). The average period has period
to days (Beach, 1943, Gomez, 1930; Picard, 1928; Reagan et al., 1954c;
been
Sahai,
1937a, b), and occasionally
or so more (Hudson, 1937a,
a
to
(Bankowski,
b;
mortality
usually possible to distinguish 1963b). forms in fowls but not in other
1
cent
outbreaks
findings
5
1961c),
100 per
published
these
4
with
asymptomatic
as
a
form of New castle disease (Beach, 1944). The occur rence of all degrees of severity, from out recognition
day
Jungherr 45
and Minard,
1944),
Osteen,
1952),
and
(Thompson
or less
on
depending
the
amount of virus inoculated
Gorrie, 1942; Farinas,
(Albiston and 1930). With aerosol
the incubation
infection,
period
shorter in warm environments in cold environments
(Sinha
Both sexes are equally susceptible (Fenetal, 1947; Rodier, 1928).
stermacher
has been
and longer
et al., 1957).
After contact exposure, the incubation period has varied from 1 to 15 days or longer (Beach, 1943; Brandly, 1959; Jungherr et al., 1946; Khan and Huq, 1963; Sahai, 1937a) with the average being 4 to 9 days (Doyle, 1927; Hudson, 1937a, b).
Breed Susceptibility There have been reports of no difference in susceptibility
Sex Differences in Susceptibility
between improved
breeds
Age Susceptibility In the
outbreaks of the less virulent form of disease,
birds
decreased
susceptibility
became
mature
(Brandly
as
al.,
et
1953; Dobson,
1939).
Similar increasing resistance with
age has
1946a, d; Brandly,
been shown towards mesogenic vaccine strains of virus (Haddow and Idnani, 1946; Nilakantan et al., 1960a). However, in natural outbreaks of the velogenic form of the disease, all ages have been suscep tible (Rao and Agarwal, 1960; Rodier,
and native breeds in West Africa (Hamil ton, 1951; Hill etal., 1953); India (Cooper, 1931; Guha and Chatterjee, 1950); Ceylon
factor
(Sturgess, 1931 ); Indonesia
castle disease and, when present, accounts
Ressang, 1961); Pakistan
for the relative insusceptibility of young chicks to virulent virus (Brandly et al., 1946c; Doll et al., 1951a) or to a lento-
(Doyle, 1948; (Khan and Huq, 1963); and the Sudan (Karrar and Mus
tafa,
among improved 1964). However, breeds, Leghorns have been found more
Island Reds and other heavy breeds (Albiston and Gorrie, 1942; Kee, 1928; Piatt, 1948).
1928; Sturgess, 1931). immunity Congenital
is
an important
in the resistance of birds to New
genic vaccine virus (Lancaster et al., 1 960).
than Rhode
susceptible
Genetic Differences in Resistance
Effect of Season of Year In India, outbreaks of Newcastle disease during the rainy
have been more common
season, and the disease has generally
sub
In an early report on this topic, Iyer and Dobson (1941a) stated that the offspring of Newcastle-resistant hens survived chal
sided during the very hot weather (Sahai, 1937a). Similarly, in the Sudan, the dis
lenge with virulent Newcastle disease virus during the first four weeks after hatching.
rainy season (Karrar and Mustafa, 1964). However, season of the year has not,
It
apparently,
seems
likely that this apparent resistance
was the result of passive transmitted
anti
body (see pages 91 to 93). More recently, Cole and Hutt (1961) found that two strains (K and C) of White Leghorns
showed significant
susceptibility
differences in
to a live wingweb
vaccine
(Figure 7). Similar genetic variation in resistance has been reported by Francis and Kish (1955) and Godfrey (1952). This variation may account for some of the differences in clinical symptoms. 46
ease has tended to appear at the end
influenced the virulence
of
of
the
the
disease in the Philippines (Farinas, 1930) or in Pakistan (Khan and Huq, 1963). In
Maryland, in the United States, cases con by virus isolation have shown a seasonal distribution, being more frequent firmed during
the winter
(Jungherr
months
and
Terrell, 1947). Inclement weather may modify the clinical course of the disease (Brandly, 1953). Greece
However, and
severe
Indonesia
epidemics
in
the
hot
during
MORTALITY DUE TO VACCINATION VACCINE
WITH A WING-WEB
Percent
7-
5
-
3 ~
1
-
Days after Vaccination (Redrawn from Cole and Hott, 1961)
Figure 7.
summer
have
been
reported
(Brandly,
1953; Mansjoer, 1961).
nervous signs at warm temperatures, more
respiratory
signs
at
cold
and
temper
However, after a study of the of Newcastle disease in Europe, Eckert (1957) concluded that atures.
epidemiology
Climatic Influences Cold,
adverse
weather
increases
the
of clinical symptoms (Brandly, 1959); and method of housing has also influenced mortality (Francis and Kish, Under controlled environmental 1955). conditions, Sinha et al. (1957) found more severity
there was no direct relationship between the incidence of fowl pest (Newcastle dis ease and fowl plague) and seasonal tem
or precipitation. Under experi mental conditions, Baetjer et al. (1960) showed that in young chicks the inhalation perature
47
of warm dry air appeared to inhibit the ciliary mechanism and this allowed a greater spread of virus in the respiratory
lone and Squibb, 1962). No pronounced change in packed cell volume, sedimenta
tract.
observed
tion rate, or buffy
coat values
has been
artificially infected
in chickens
with Newcastle disease virus (Bierer et al., 1963).
Routes of Infection Chickens
In
3 to 6 weeks
most susceptible
old have been
with New
to infection
of Newcastle
a severe outbreak
in
ease
6-week-old
chicks
reduction
significant
there
in the blood
A
dis
was
a
serum
castle disease virus by the intramuscular
levels of total protein, vitamin
or respiratory routes, and most resistant to
carotenoids, but the ascorbic acid level was
virus administered by the alimentary
raised (Squibb et al., 1955). In artificially infected chicks, an increase in riboflavin
route
(Kohn, 1955). The upper respiratory tract seems to be the site for the early multipli cation of the virus following intranasal infection (Maestrone and Coffin, 1964). In reviewing the topic of portals of entry, Beaudette under may
(1943) has emphasized
field
conditions,
represent
per os infection
of
entrance
the
through
either
the digestive
piratory
tract.
Further reference
virus
or the res to this
subject is made under the heading Vaccines"
that,
"Live
on page 97.
of the ration has reduced
supplementation mortality
10 to 17 per cent (Squibb,
crease over the normal per
cent
of
requirements
of vitamins has resulted in in
increase
a
a
13
disease
Newcastle
mortality.
of an in
Following the administration activated
Newcastle
disease
vaccine
pullets aged 12 weeks, total cholesterol been determined egg yolk (Schiavo, ments, vaccination
Effect of the Virus on Avian
1963). an appreciable in
By way of contrast, number
and total
to has
in samples of serum and 1963). In these experi had no effect on choles
terol levels.
Physiology has increased nitrogen retention during the
The rise in body temperature in groups of vaccinated chickens, subsequently in fected with virulent virus administered by
incubation
the subcutaneous
Infection with Newcastle
retention
disease virus
stage and depressed
during the clinical
nitrogen
stage
(Sans-
routes,
and intravenous
has been examined by Mantovani (1949).
THE DISEASE IN CHICKENS Velogenic Form
sion, increased rate of respiration, sive weakness
Symptoms
and prostration.
progres
There
is
usually an early rise in temperature (Doyle,
In the velogenic form, Newcastle disease and spreads rapidly appears suddenly
1935; Guha and Chatterjee,
1950; Hud
through a flock. Peracute cases have been
son, 1937b) which increases on the fifth or sixth day to a peak of 4° to 6°C above
reported in which birds were found dead
normal.
without
(1961a) found that temperatures which remained higher than 42°C for
(Sahai, 1963). ness is 4S
having
shown
any
symptom
1937a; Iyer, 1943; Khan and Huq, In more typical cases, initial dull followed rapidly by marked depres-
In
artificially
infected
birds,
Squibb more
than 48 hours,
below normal,
and then dropped
were associated with acute
Figure 8. — The velogenic form of Newcastle disease — symptoms of paralysis. Dr. G. L. Bannister, Animal Diseases Research Institute, Canada.)
neurotropic involvement or death. The markedly subnormal temperature before death is a characteristic 1931; 1946; mon
Hudson,
symptom (Cooper,
1937b;
Jungherr
et
al,
1937a). Diarrhoea is com in the early stages of the disease. Sahai,
Fae;es are usually profuse, watery, green ish or yellowish, and occasionally blood 1942; stained (Albiston and Gorrie, Farinas, 1930; Rodier, 1928). Rate of respiration
is
increased,
bird shows a characteristic
and usually
the
prolonged gasp
ing inspiration with outstretched neck and head and partially opened beak. In arti ficially infected fowls, gasping respirations may be absent (Acevedo,
1933), but there
(Courtesy of
Guha and Chatterjee (1950), Hilbrich (1963) and Iyer (1943). Distension of the crop with gas or sour fluid is common.
There may also be a dis charge of mucus from the mouth or nos trils (Johnstone, 1933; Kylasamaier, 193!;
Rodier, 1928), and sometimes from the (Kylasamaier, 1931). However, not many of these symptoms were noticed in the 1932 outbreak in Australia (Albiston and Gorrie, 1942) or in outbreaks report ed by Gordon and Asplin (1947) and Fabricant (1950). There is usually rapid dehydration and cyanosis of the comb and wattles; and eye
occasionally
oedema
around
the
head
(Albiston and Gorrie, 1942; Hudson, 1937b). Photo graphs of affected birds which show many
(Thompson and Osteen, 1952). Birds that survive the acute phase of the disease often show involvement of the
of the above symptoms appear in the pub
central nervous system (Ressang, 196!) by weakness or paralysis of the legs and,
may
be
a
sharp
cough
of Doyle (1927), Albiston and Gorrie (1942), Beach (1943), Brandly (1959), Fritzche and Gerriets (1962), lications
occasionally,
the wings (Figure 8).
Other symptoms include clonic spasms. 49
Figure 9. — The velogenic form of Newcastle disease — lesions in proventriculus. (Courtesy of Dr. G. L. Bannister, Animal Diseases Research Institute, Canada.)
muscular tremors, torticollis, emprosthotonos,
and
opisthotonos,
abnormal
move
perature
was
mortality
100
whereas at a low environmental
cent;
per
tempera
ments such as walking in circles. Opacity
ture it was 55 per cent. Using different
of the cornea has also been reported (Abrams, 1961; Hill et al., 1953; Thomp
management
son and Osteen, 1952). Egg production falls abruptly;
found that chickens
or imperfectly
Effects on the production eggs
and soft
shelled eggs may be laid.
are more characteristic
and quality
of
of the meso-
practices,
(1955) obtained Newcastle
Francis and Kish
contrary
results:
they
exposed to a virulent
virus in a warm battery room
had a much lower mortality
than similar
groups in colony houses.
The duration of illness has been report
genic form of the disease and are described
ed as one to two days (Sturgess,
later under that heading.
three to four days (Iyer, 1943; Kylasamaier, 1931; Rodier, 1928) and ten days
In
the
velogenic
form, mortality
is
90 per cent. Occasionally, pens of older birds have survived field
over
(Dobson, 1939). The influence of environmental
infection
tempera
ture was examined by Sinha et al. (1957) who found that at a moderately high tem 50
1950). Surviving birds seldom fully recover (Iyer,
(Guha and Chatterjee,
Mortality usually
1931),
paralytic
1943; Rodier, 1928).
Macroscopic
Lesions
Post mortem lesions in several hundred infected
birds
have
been
described
by
Culzoni (1949), Doyle (1927), Guha and
Figure
70. — The velogenic form of Newcastle disease — lesions in intestine. Dr. G. L. Bannister, Animal Diseases Research Institute, Canada.)
Chatter jee (1950), Iyer (1943), Jungherr et al. ( 1 946 ) , Kaschula ( 196 1 ) and Picard (1928). The post mortem findings des
cribed in 23 published reports have been reviewed by Beaudette ( 1 943 ) .
Lesions acute
cases
are usually than
in
more
definite
in
chronic cases; but
(Courtesy of
there is no single lesion which indicates the severity of the clinical disease (Jung herr et al., 1 946) . In some outbreaks there has been a general absence of gross patho logical changes (Andrews, 1948; Cooper, 1931; Hill et al, 1953; Sahai, 1937a); while
in others the presence of a simul 51
THE YELOGENIC FORM OF NEWCASTLE DISEASE DISTRIBUTION OF LESIONS Diagram of the sites of the
12
-
lesions
seen regularly in the alimentary tract (Redrawn from Kaschula 1961)
Figure 11.
taneous tapeworm ently inhibited
infestation
has
appar
of intestinal
the formation
lesions (Gavey, 1958). Lesions are mainly haemorrhagic inflammatory.
petechial and are commonly
and
are usually
Haemorrhages
found in the
breaks
have been associated
with diph
theritic lesions of the mouth which began circular, yellowish elevations as small,
(Orr and John, 1946). In other outbreaks the
cloaca
has
been
highly
congested
(Kuppuswamy, 1935).
and submucosa of the proven(Figure 9), gizzard and intestinal tract (Figure 10); on the trachea, epicar-
with the lymphoid follicles of the intestine
dium,
and caeca are common. These lesions may
mucosa
triculus
branes;
pleura,
mesentery,
and in the general
Less frequently, ulceration
sac mem
musculature.
severe haemorrhages the
and
2
inches
occur
necrotic difficulty
pharynx
may
raised
or diphtheritic
and
haemorrhagic
lesions associated
or cover areas The 1931). removed only with
be very small haemorrhages
(Hudson, 1937b). In Malaya, initial out52
in
air
Bluish-red, necrotic
long
(Sturgess,
membrane
is
and leaves an ulcerated
area. In
less acute cases these ulcerative
lesions are
more common. The development
and dis
tribution of lesions in the alimentary tract (Figure 11) have been described in detail by Culzoni (1949), de Kock (1954), Guha and Chatterjee (1950), Hecke (1943-44) and Kaschula ( 1961 ) .
In
acute
the spleen
cases,
been
has
(Kaschula, 1961; Ressang, 1961; Schoening and Osteen, 1 948 ) .
enlarged
Microscopic Lesions Results of the examination experimental
cases
of over
1 60
have been reported by
Jungherr et al. (1946), and of a larger number of field cases by Culzoni (1949) and Ressang (1961 ). These authors found that lesions gall
in the spleen, intestine, liver,
bladder and heart were essentially
exudative lesions
necrotizing
in character. Splenic
have been more severe when the
incubation period has been short (Fuku1934). In the lungs,
shima and Shimonura, central
nervous
system and eye,
have been mainly proliferative
lesions
and hyper-
aemic (de Kock, 1954; Thompson and Osteen, 1952). In one series of specimens, the lymphocyte was the cell type pre
Stubbs (1946). Typically, in a susceptible flock, this form appears suddenly and spreads
symptoms
usually
symptoms
may
chial
haemorrhage
histological
has
examination
been
(Obel
seen
on
et
al.,
1956). In immune birds exposed to viru lent virus, focal fibrosis in the air sacs and lungs has been described
(Jungherr
etal, 1946).
Mesogenic Form Symptoms
A
general account of the symptoms and
lesions associated with the mesogenic form
of Newcastle disease has been given by Beaudette (1951b), Morgan (1946) and
subside then
and
nervous (Stubbs,
appear
1946). The latter are of variable incidence (Pomeroy and Fenstermacher, 1948). Involvement of the central nervous system (Figure 12) is more common in young chicks than in older flocks (Abrams, 1961 ; Baker and Hays, 1947; Beach, 1943). In some flocks over 3 months of age, nervous symptoms
have
not
been
recognized
(Beach,
1946). Effect on egg
largely
on
the
production
depends
of the disease, the stage of the
severity
individual susceptibility,
laying period and whether or not moulting occurs (Schoening and Osteen, 1948). Thus, Biswal and Morrill (1954) found that in some birds the pause in production was 7 days and in others 22 days. In one
another,
and Osteen, 1952).
(Beach,
Morrill, 1954). Some birds have greenish or yellowish diarrhoea (Zuydam, 1950b). Within two or three weeks, respiratory
outbreak,
In artificially infected fowls, severe bron
respiratory
gasping
1943). There is a marked drop in appetite. Egg production falls and may stop on the ninth day (Beach, 1943; Biswal and
1951a). Extensive oedema has been seen in the liver (Hill et al., 1953) and cornea (Thompson
show
and
coughing
dominantly involved in brain lesions dur ing the acute phase of infection (Karzon and Bang,
Birds
rapidly.
distress,
egg production was adversely
affected for 12 weeks the
birds
(Knox, never
1950); in
returned
to
(Piatt, 1948). Accord ing to De Moulin (1951 ), decrease in egg normal production production
results in part from a degenera tion of the pituitary gland. A feature of the mesogenic form is its effect on egg quality. During the early stages of the disease, the shell is often discoloured, or missing; and imperfect many eggs are abnormally shaped. After recovery,
the
appearance
usually returns to normal;
interior quality
and
weight
of
may
be
of the shell but changes in
reduction a
more
in
egg
permanent
nature (Knox, 1950; Quinn et al, 1956). There is often an accumulation of small air bubbles instead of the true air cell. The main features include decrease in albumen 53
quality (Lorenz and Newlon, el
al,
1956), deterioration
1 944;
Quinn
in keeping qual
ity, and increase in the percentage of stuck
sist for a long characterized
time and which is often
by nervous
phase of humoral
yolks. Quality and weight of the yolk are
immunity
not altered. In some outbreaks, the weight
haemagglutination-inhibition
and thickness
of the shell of
eggs
laid
during the recovery phase have decreased; however, the albumen rarely decreases in weight (Biswal
and Morrill,
1954).
Course and Mortality Woernle and Siegmann (1954) and Woernle (1955) have described four dis tinct phases in the course of Newcastle disease: (1) phase of panagglutinin forma tion or the appearance of auto- or heteroand Siegmann, (Woernle 1954); (2) phase of viraemia; (3) phase of virus and antibody balance which may per agglutinins
which
antibody is readily
symptoms;
(4)
formation and revealed by the test.
Mortality in the mesogenic form is con siderably
lower than in typical
outbreaks
of the velogenic form. Mortality may vary from 5 to 50 per cent in mature flocks and may exceed 50 per cent in young and Hayes,
(Baker
etal,
chicks
1947; Fenstermacher
1947; Morgan, 1946; Schoening
and
losses 1948). In one outbreak, were in the ratio of five paralysed birds for
Osteen,
each dead bird (Goldhaft and Wernicoff, 1948). Under experimental conditions, an increased mortality associated
with
serves (Squibb,
rate in chicks has been
depleted
vitamin
A
re
1961b).
Figure 12. — The mesogenic form of Newcastle disease — nervous symptoms. (Crown Copy right. Reproduced by permission of the Controller of Her Majesty's Stationery Office, London.) 54
Macroscopic Lesions
with
There is considerable findings
(Sinha (Asplin
genicity
of
tropisms
Also,
in the
variation
al, 1952), post mortem et al, 1949) and patho
et
mesogenic strains of the virus.
haemorrhagic
and
inflammatory
lesions vary greatly between flocks and, to some extent,
between geographical areas (Jungherr and Markham, 1962; Ressang, 1961). Thus, an Italian strain has produced lesions confined mainly to the digestive an American strain has
system; whereas
mainly in the respiratory
caused lesions
and nervous systems (Mantovani et
al,
1954).
The
average
of
incidence
intestinal
lesions produced by four European strains of the virus in over 1,000 individual was 63 per cent (Jungherr et al.,
chickens
caseous
exudate
tropic characteristics. the
by
These are indicated
of vesicles
development
of the
clouding
eyes
the aqueous humour (Thompson and Osteen, 1952). In the mesogenic form of the disease, matory
cells
in
the spleen is usually small, pale, anaemic, and mottled
in appearance.
The kidneys often show inflammation evidence of nephritis. Microscopic
cause inflammation
strains of virus
and cellular infiltra
lesions are mainly proliferative
per cent, 1949b). chickens
72 per cent, heart 52
various
In
40 per cent
contrast,
conducted
(Lucam,
of 800
in a study
Jungherr
by
et
al.
(1946), five American strains caused gross lesions in 24 per cent of cases. Kohler (1953) has reported that 56 per cent of the cases he examined showed no intestinal
lesions post mortem.
macroscopic
Haemorrhages
are
common
in
the
of the proventriculus and less common in the intestine. In some out
mucosa breaks,
enteritis
there
has been severe catarrhal
(Kawashima
et
al.,
1953);
in
others, there has been fluid or mucus in (Watanabe et al, 1952). The more pneumotropic strains of virus often the trachea
of the air sacs which a film of yellowish exudate (Beach, 1943). This inflammation of the air sacs shows a marked tendency cause a cloudiness may
develop
to progress
into
and, in combination
E. coli infection, carditis,
lesions
perihepatitis
with
an
of severe peri
and
airsacculitis
of the serous
thoracic
and
Lesions
The more pneumotropic
breaks of Newcastle
72 per cent, cloaca
However, in
acute cases, enlargement of the spleen has been observed (Watanabe et al., 1952).
tion
disease in France was follows: liver 74 per cent, proventriculus
on the
1959) or by a caused by inflam
wattles and comb (Brandly,
of 1946). The percentage distribution lesions in different organs in natural out as
(Gross,
result
1961a, b). Some strains of the virus have dermo-
and
membranes
abdominal
cavities.
of the Lung
and exuda tive in character (Brandly, 1959). In some specimens, atelectasis has been the only abnormality Secondary
noted (de inflammatory
Moulin, changes
1951). in the
pulmonary and abdominal air sacs are characterized by oedema, cellular infiltra tion and fibroblastic proliferation in the more chronic cases. Mild congestion and mucous
exudation is often present in the trachea. Catarrh of the bronchi has also been reported (Jungherr and Minard, 1944; Mochizuke et Inflammation and stromal
al,
1952).
of the ovary with oedema
vacuolation
cribed by Biswal and
have been des
Morrill
(1954) and by Pasley and Auer (1958). Hypertrophy and hyaline necrotic foci have been present in the spleen of many specimens (Jungherr and Minard, 1944).
In the central nervous system, virulent strains have caused extensive hyperaemia and proliferation of the endothelial
cells
(Jungherr and Minard, 1944), and mild or severe degenerative changes of the neurones and ganglia (de Moulin, 1951; Jungherr, 1963; Sullivan, 1958). Lesions 55
of encephalomyelitis observed in the early following vaccination, have general ly disappeared within five months (Salyi and Hodosy, 1 952). The brains of chickens stages
nervous
showing
have
symptoms
been
examined with the object of localizing the main
histological
(Auer, 1952;
changes
Karzon
and Different Bang, 1951a). lesions appear to be associated with vari ations
in
with strains of
observed,
the symptoms
typical American and European virus
in the histological
differing
they produce (Potel, paralytic
lesions
Kohler
(1953),
examination
microscopic
who considered of the central
nervous system a valuable aid in diagnosis,
found
a non-purulent
encephalitis
in 88.4
per cent and a neuritis in 72.9 per cent of the cases
Similarly, Mits-
he examined.
cherlich
et al. (1953) have reported that 89 per cent of the cases they examined
were diagnosed
and 80 per
histologically
cent serologically.
In the pituitary gland, the virus has caused a degranulation of acidophile and basophile cells (de Moulin, 1951; Pasley and Auer, 1958). In the intestines, lesions involving proliferation of the reticuloendothelium
have
observed
been
gerle, 1952). The haematological
(Zan-
picture
in
the
cases, Chandrasekharan
10 spontaneous
and Krishnan (1958) observed a slight rise in the total red blood cell count and a reduction differential reduction
in the white cell count. count,
was
there
in lymphocytes
increase in heterophiles.
a
In the marked
and a significant
In contrast, Ishii
and Kobayashi (1952) recorded a marked temporary leucocytosis during the febrile stage.
Weidenmuller (1960) found eosino-
philia, monocytosis
and lymphopenia
suggested that lymphopenia as a diagnostic
criterion
when serological 56
tests
and
could be used
in the early stages were still negative.
ob
after infection immediately which lasts 24 to 96 hours, and the second
one
starting on the 1 8th day which lasts seven days (Machado, 1951). On the other hand, Pehl (1959) concluded that the cell picture in bone marrow and circulating blood is not sufficiently characteristic
to distinguish
disease from infections
Newcastle
caused
by other agents. Furthermore, he thought that the leucopenia
which occurs in New
castle disease is not diagnostic.
Lentogenic Form Symptoms This form of the disease is characterized by mild respiratory symptoms and by a sudden drop in the egg production of lay ing flocks. There is usually depression
of appetite.
impairment breaks,
In
involvement
respiratory
and
some
out
is very
(Asplin, 1952; Asplin et al, 1949) and is only detected when the birds are roosting. In other instances, there may be no respiratory symptom (Kutlesa, 1952). mild
There
are usually
no nervous
symptoms
(Abrams, 1961), and egg production re turns to normal within a few weeks. How ever, in some outbreaks nervous symptoms have predominated (Salyi et al., 1955). The course of the disease varies, but usually one
mesogenic form of the disease has varied.
In
served:
1950). However, in a Karzon and Bang
chicken,
(1951a) found that brain lesions were not related to the virulence of the infecting virus.
Two periods of leucopenia have been
to
there is complete eight
weeks,
degree of debility.
In
on
young chicks,
discharge and abnormal respiratory are common.
Mortality In adult fowls mortality but it may reach
within
recovery
depending
is
the
nasal sounds
negligible,
50 per cent in young
chicks.
Macroscopic
Lesions
Haemorrhagic absent.
A
and visceral
mild tracheitis
lesions
are
may be seen in
early cases and in young birds.
Microscopic
A
Lesions
disseminated
been found
has encephalomyelitis in a field outbreak (Kutlesa,
1952). In a study of vaccinated chicks, Auer (1953) found no histological evi dence of degeneration in nerve cells. How ever, there was an inflammatory reaction in the central nervous system. Vascular engorgement and infiltration with lym phocytes have lasted 14 days after vaccin ation with no clinical symptom being seen during this period (Auer, 1954). Gross (1963) examined microscopic lesions in air sac membranes, lungs and trachea of 3-week-old susceptible chickens exposed to an aerosol of the Bl strain of virus.
In
the air sac membranes there was
disease and some of which did
Newcastle
not, have already been mentioned on page 33.
Thus, Newcastle
disease
clinical signs (Reid, 1961). Also, Minard and Jungherr (1944) have reported high levels of neutralizing antibody in a flock in which there was no evidence of the disease. This was understood to indicate In some cases, spread a latent infection. of an inapparent form of the disease (Jung herr and Terrell, 1946) has been thought to result from carriers of the virus. In one instance, inapparent
infection
had existed
a progressive increase in connective
tissue
on a large breeding establishment
for about
there
years
16 days post vaccination;
after, the walls of the air sacs became thinner. Lymphoid infiltration was marked in the air sac membranes and in the lungs around the parabronchi, but became more localized in the tracheal epithelium. It was concluded that the Bl strain had resulted in lesions similar to those associated with infectious bronchitis virus or Mycoplasma gallisepticum infection.
for two appeared
1951).
Bankowski (1961c) has emphasized that disease may Newcastle asymptomatic often be diagnosed only by chance. He has out that the haemagglutinationand
pointed
inhibition titre fluctuates considerably outbreaks
that inapparent
may be missed
of sera are However, such variations in
sufficient
unless
examined.
are not
numbers
to asymptomatic
confined
marked
infections:
Reports of asymptomatic infections, some of which resulted in the spread of
clinical evidence
before
(Beach,
titre
Asymptomatic Form
been
has
in the absence of
serologically
diagnosed
variation
in
HI
titre
in the mesogenic
has also been observed
form of the disease (Fabricant, 1950).
THE DISEASE IN TURKEYS Symptoms
reported
In
the mature turkey, there may be mild respiratory symptoms. Nervous symptoms are seldom
et al., (Fenstermacher 1946) but have been reported occasionally (Walker, 1948). In laying flocks, egg pro seen
an outbreak
in turkeys
which
and largely in a subclinical form. Respiratory symptoms were absent, spread slowly,
but partial or complete paralysis of one or both legs was common.
duction has dropped almost to zero. Eggs
Course and Mortality
have been soft shelled and deformed
After outbreaks in some breeder flocks, fertility and hatchability have returned to
have had watery 1961 ) .
albumen
(Gale
et
and al.,
Fertility and hatchability have been
reduced.
In flocks of young poults, respiratory symptoms, appear
depression
early.
Gray
et
and al.
inappetence
(1954) have
normal (Gale et al, 1961). Death may occur in mature birds (Fenstermacher et
al,
1946);
and
in
some
outbreaks
the
disease has been peracute with high mor tality
(Gordon
et
al.,
1948;
Walker, 57
1948). In young poults, mortality of 60 per cent or higher has been observed ( Fenstermacher
et al,,
reported by Gale et al. (1961) who noted that in some outbreaks there were lesions in
1946).
Macroscopic Lesions Gross lesions are rare, but some petechiae on the heart and a cloudy appear ance of the air sacs have been reported.
the
of experimentally
and naturally
turkeys
infected
has been
nervous
central
and capil
in the respiratory
lary congestion
system, in the
and extensive neuronal degeneration cerebrum, other
brain stem and cerebellum.
In
with
an
outbreaks,
occasional
Microscopic Lesions The histopathology
and
respiratory
systems: extensive haemorrhage
(Gray
the
al,
et
gliosis
perivascular in
observed
focal
cuffing
brain
and
has been
spinal
cord
1954).
THE DISEASE IN DUCKS AND GEESE Although these species are more resist ducks and geese have been reported
100 per cent (Bush,
have been observed
depression
(Albiston
and Gorrie, 1942). Elsewhere, ducks and geese have undergone symptomless infec
In England, fowls have
tion (Khan and Huq, 1963). ducks exposed to infected remained
usually
apparently
healthy
(Asplin, 1947). In Haiti, ducks and gos
of ducklings was
in one flock
mortality
(see page 29). In Australia, locomotory dis turbances, abnormal gait and general
to the disease, and
lings have succumbed
ant than turkeys, natural outbreaks among
experimental
1954). In contrast, the inoculation of mature ducks
with virulent
Newcastle
serum
disease virus has
in no clinical symptom
resulted
has shown
no
virus
and the
neutralizing
(Iyer, 1945). In a few outbreaks, some ducks have died (Andrews, 1948). Post mortem findings have been generally antibody
and
(Albiston Asplin, 1947). negative
Gorrie,
1942;
THE DISEASE IN GAME BIRDS Natural outbreaks tality
have
guinea-fowl
been
with
reported
and partridges.
the disease on these birds
varying
mor
has been re
death occurred
five days after the appear
58
the serous
have been reported
(Parnaik and Dixit, 1953).
studied
that,
on
disease
(1958)
have reported
(Perdix perdix), a significant lesion consisted of numerous microscopic ulcers in the intestine which appeared to the unaided eye as very small white foci (Thompson, 1955). In other outbreaks, coat of the gizzard
Laroche
The effect of
partridges
haemorrhages
and
Newcastle
viewed by Beaudette ( 1943 ) and Brandly (1959) (see page 28). In one outbreak in
prominent
Lucas
in pheasants,
ance of symptoms.
in the partridge in the
acute
Post mortem
were those of enteritis.
and
form,
findings
In the chronic
form
of the disease, most partridges made a slow recovery. A strain of virus isolated from a natural
infection
in a partridge
has been
identical to strains isolated from chickens
(Placidi and Santucci, 1953c). A natural infection in guinea-fowl has resulted in symptoms of encephalitis (Placidi and Santucci,
1953c).
METHODS
DIAGNOSIS BY SEROLOGICAL
The
et al., 1946d). Determination of the end titre in the HA titration of virus,
Brandly
(HA Test)
Haemagglutination
haemagglutinating
property
of
disease virus has been described
Newcastle
by Burnet (1942) who found it capable of agglutinating the red blood cells of man, guinea-pig, mouse, fowl, sparrow and frog of human, (Hyla). The agglutination bovine and avian spermatozoa by New castle
disease virus has been described by
similar to the agglutination of red blood cells. Similarly, the agglutination of leucocytes and blood (Jerushalmy platelets has taken place of al, 1963). The agglutination et Chu
(1953)
avian
red
being
as
blood
cells
by
Newcastle
virus can be inhibited by a sub stance extracted from human and porcine lungs (Rice and Stevens, 1957). Similar inhibiting substances have been found in sera from humans and Rhesus monkeys disease
(Wenner et al, 1952b),
in the allantoic
fluid of normal 13-day chicken
embryos
(Williamson et al, 1955) and in normal human urine (Tamm and Horsfall, 1950). In fowl serum, the removal of a nonspecific inhibitor has resulted in an in titre (Traub, crease in the neutralization 1956). Similarly, the removal of an ethersoluble
antigen
covering
the
virus
has
resulted in an increased haemagglutination sensitivity
and a higher titre in the
HI
test
(Larski,
1962). Ether disrupts the virus particles, but the ease of disruption varies
strain to strain (Waterson and Cruickshank, 1963). The virus has failed to agglutinate the cells of monkey (Af. irus), ferret, sheep, from
horse,
rabbit,
thopsis)
marsupial
(Smin-
and tortoise. However,
the virus
the erythrocytes
of camels
can agglutinate
(Hashmi and Hasnain, Santucci,
mouse
1954b; Placidi and
buffalos
and
1956) (Hashmi Hasnain, 1954a), cattle (Cordier et al, 1951; Ozawa and Chow, 1958) and other species of animals (Abdel et al, 1960;
has
using bovine and equine erythrocytes, been
both
and
time-consuming
difficult
(Ozawa and Chow, 1958). All strains of virus are not equally active mammalian
against
erythrocytes
concluded
(1950)
agglutinate
(Mac-
1956). Winslow et al. that the ability to
pherson and Swain,
erythrocytes
of certain mam
malian species differed between strains of
It
the virus.
was thought that this property
could be used to identify different strains.
Girotto
(1954) found that two Italian
strains did not agglutinate horse red blood cells whereas other strains did. Red
cells
shown
of different
variation
chickens
by Newcastle
agglutination
have
in their susceptibility
to
disease virus
(Bang and Foard, 1952; Brandly et al, and Gray, 1962). Con versely, different strains of virus have shown variations in agglutination (Beach, 1947; Cochrane
and some strains have been non-
1948),
of red blood cells (William
agglutinators son et
al,
strains,
1955). With highly pathogenic a minimum concentration of
approximately
et al, 1947). With a the haemagglutination
glutination
(Hanson
lentogenic
strain,
activity
LD50 of virus to produce haemag
10-6 embryo
has been necessary
has been destroyed
after several
56°C (Torlone, 1956). For many strains of Newcastle disease virus, at the resistance of the haemagglutinin 56°C has been a relatively constant and minutes
at
characteristic a means
feature and it might serve as
of distinguishing
between strains
or 1963) (Nitzschke mutations of strains (Goldman and Han son, 1955; Hofstad et al, 1963). Further and
Schmittdiel,
more, heat has had less effect on the ability
of some strains
to
agglutinate
chicken
infectivity for chick embryos (Liao et al, 1953). How cells than it has on their
ever, no conclusive
relationship
has been 59
demonstrated between the immunogenicity
disease virus,
of the haemagglutinating property of substrains of New castle disease virus (Hofstad et al, 1963b).
particles
and the heat sensitivity
Newcastle disease virus first agglutinates chicken
red blood
cells by causing
the
formation of a lattice-like matrix and then elutes from the cells (McCollum et al, 1957). Other characteristics of haemagglutination have been reported by Hirst (1950). The virus appears to be adsorbed more completely and to elute less rapidly from chicken red blood cells at 4°C than at room temperature (Florman, 1947; Sagik and Levine,
1957),
giving
clear
a
HA
reaction with certain strains (Andrewes et al, 1955; Berke and Golem, 1950) but not with a vaccine strain (Bang and Foard,
Bohm and
Nevertheless,
1952).
(1961), who made comparisons different
temperatures,
found
end-points
were obtained
peratures,
and
HA test should
at
four
that higher
at higher
concluded
they
Espig
tem
that the
be carried out at room tem
small
a
permanently
of virus on the red
number remain
blood cells. Such sensitized red blood cells can be lysed and stored indefinitely out
of antigenicity
loss
with
(Geurden
and
Devos,
1955). Specific antisera against Newcastle
ease
virus-treated
chicken
dis
red blood
cells
has been prepared (Gardner et al, 1954). However, Swain (1959) found a serum factor which reacted with Newcastle ease
virus-treated
red blood
dis
cells in the
serum of normal animals of many species. This serum factor appeared to be distinct from specific antibodies to Newcastle dis ease virus. Previously, Burnet and Ander son
(1946)
human
cases
had
shown
of infectious
that
sera from
mononucleosis
would agglutinate human red blood sensitized by the action of Newcastle
cells dis
ease virus.
Modification of the virus antigen potassium elution
periodate
property
Libert, 1952).
(KI04)
by
destroys the
of the virus (Bang As a result, the HA
and test
perature. However, the haemagglutination
can be read at any time up to 12 hours;
inhibitor present in normal allantoic fluids room temperature than at 4°C (Williamson et al, 1955). It has been shown that Newcastle dis
whereas, with unmodified
is less active at
ease
virus
and
to
is adsorbed in
platelets
(Jerushalmy
et
al,
tion of the virus
to red blood a
1961).
cells
similar manner However,
elu-
from platelets is much
slower than from red blood cells. Bang and Libert (1949) have reported red blood cells sensitized by
cerine to virus suspended in 0.1 per cent
that chicken
formol
Newcastle
haemagglutinating
antigen (Cabasso
by sera which did not agglutinate normal red cells. Thus, sera taken from chickens
1951). A non-infectious
haemagglutinin
five to seven days after inoculation
particles
virulent
disease virus were agglutinated
Newcastle
nated sensitized blood
with
cells.
disease virus aggluti cells but not normal red
This haemagglutination
of
saline
the et
al,
whose
to the in
cribed (Rott et al, 1962). Treatment with ether has resulted in separation of the
from an components American strain of virus but not from an Italian strain (Rott et al, 1961). Dialysis of allantoic fluid or repeated freezing and
ment of red blood
thawing
60
stabilizes
complete forms of the virus has been des
haemagglutinating
Newcastle
markedly
appear to correspond
sensitized red blood cells facilitates rapid diagnosis (Geurden and Devos, 1955). In studying this type of reaction, Anderson (1947) concluded that, following treat cells with
com
antigens,
of the virus from the red within two hours cells occurs blood (McCollum et al, 1957). When inactivated by heat or formalin, the virus retains its ability to agglutinate red blood cells (Berke and Golem, 1950; Bodon, 1953; Brandly et al, 1946d; Burnet et al, 1945; Walker, 1952). The addition of 50 per cent gly plete elution
has reduced
the haemagglutina
(Vrtiak et al, 1959). The liber units by ether haemagglutinating
tion titre ation
of
in the identification of virus humour of naturally or
been utilized
in the aqueous
has produced a suspension of spherical particles of about 30 micron diameter with long filaments (Sokol et al, 1961 ) . Ultrasonic irradiation of Newcastle disease virus has resulted in loss of virul ence but an increase in its haemaggluti nating property (Garay and Syent-Ivanyi,
artificially infected chickens (Clark et al, 1955, 1957; Topolnik, 1957) but, in
1955).
tracts for this purpose have been prepared
treatment
Using
high-speed
of
pensions
of
centrifugation
Newcastle
disease
sus
virus,
al (1950) demonstrated the of two haemagglutinating par ticles of different sizes. The larger appeared Granoff
et
presence
to be the infectious particle
principle;
the smaller
appeared to be non-infectious.
In
studies, Granoff and Henle subsequent (1954) showed that the larger component was readily adsorbed on to red blood cells in the cold, whereas only small amounts of the smaller
particle
combined
with
the
cells. Haemagglutination
test techniques
have
(Anon., 1946b; Berke and Golem, 1950; Chu, 1960; Cun ningham, 1960; Fabricant, 1949a; Kaplan, 1949). A micro-method of conducting the test, described by Takatsy (1956), was based on the use of a 0.025 ml. loop instead of a pipette, together with a metal been described
in detail
dropper also calibrated to deliver 0.025 ml.
The incubation time reduced
if
for the test was
the plastic plates containing
the
for 10 seconds. Possible sources of error and factors influencing HA test results have been dis cussed by several authors (von Sprockhoff, 1961; Anon., 1959; Bang and Foard, 1952; Burnet et al, 1945; Brandly et al, 1947; Cunha et al, 1947). The standard HA test involves the pre paration of dilutions of the virus antigen in constant volumes of a red blood cell suspension. To facilitate diagnosis, rapid slide or plate tests have been made in which only one dilution of the reagents is used (Walker, 1952; Zargar and Pomeroy, 1949). The haemagglutinating property has reagents were centrifuged
general, results have not been encouraging
for diagnosis (Baldwin, 1962; Scott
et
al,
1956).
Rapid diagnosis using tissue extract in an
HA
test
has been studied. Tissue
ex
from lungs (McClurkin et al, 1954), liver and spleen (Monti, 1952). The method may be of value in the early stages of infection
before
serum
antibodies
have
appeared. However, Belic
(1962) has re ported a high percentage of negative HA reactions and has concluded that such are of limited value. or tryptic digestion of the extract have been found to increase the specificity of the test (Berke and Golem, 1950). On the other hand, the use of fowl red blood cells sensitized by suspensions of infected rapid test procedures Peptic
material
organ
has been a reliable
pro
cedure (Grausgruber, 1958).
Haemagglutination-lnhibition (HI Test) Burnet (1942) and Lush (1943) showed that serum from fowls that had recovered
from the disease contained antibody cap able of inhibiting the haemagglutination of red blood cells by the virus. The haemagglutination-inhibition
(HI)
useful
as
provided qualitative
quantitative
test has
well
as
data.
Using antigens and sera produced from nine different strains of Newcastle virus,
Siegmann
showed
Woernle
that the homologous
hibited
haemagglutination
dilutions tan et
and
than heterologous
al
disease
(1955)
serum
in
in far greater sera. Nilakan-
(1962) compared various pro tein fractions of plasma from Newcastle disease immune chickens and showed that the
euglobulin
amount of body
—
HI
contains
the
maximum
and virus-neutralizing
anti
no antibodies were demonstrated
in the albumen fraction of the plasma. 61
for the
Two procedures been used.
In the
HI
have
glutination
(Anon.,
Newcastle
test
alpha procedure
1946b; Fabricant, 1949a) the virus sus pension is diluted serially and mixed with equal volumes of the serum under test. In the beta procedure (Anon., 1956; Chu, 1960; Gentry, 1957) the serum is diluted serially and mixed with a constant amount
of virus dilution number of
containing
HA units. A
a
comparison
alpha and beta procedures
known of the
has been made
by Brandly et al. (1947). Electron microscope studies of the test
were
Brueckner
conducted
(1953)
by
who
seeing virus-like particles
Reagan have
HI and
reported
adhering
to the
Horsfall,
of chicken red blood cells by virus (Ginsberg and Thus, fresh normal 1949). disease
human sera have caused the virus to lose its capacity to agglutinate red cells (Bang et al, 1951 ) ; though this phenomenon has been influenced by the unsuitability of the red blood cells of certain chickens (Ander
son, 1948). The test has been found
to be of little
in the acute form of the disease (Scott et al, 1956; Valadao, 1955) ; and in early infections it should be supplemented value
by tests for detecting circulating virus (Topolnik and Hallauer, 1950). It has been suggested that the strain of invading virus (Doll et al, 1950d) and the genetic background of the chicken affect HI titre. Some chickens can apparently overcome
of agglutinated red blood cells. No virus-like particles were evident in the presence of immune serum. Details of HI test techniques have been given by several authors (Anon., 1946b, 1956, 1959; Chu, 1960; Crawley, 1954b; 1960; Doll et al, 1950d, Cunningham, 1951d; Fabricant, 1949a; Kaplan, 1949). Takatsy (1956) has described and illus
1950b). disease, some chickens
trated a micro-method.
tion have remained negative to the
surface
of absorbent
Discs
paper
(serodiscs) have been used to collect samples of whole blood for subsequent elution and testing by
(Andrews, 1963). This resulted in the identification of
the beta procedure technique
85 per cent of pheasants with a serum titre of 200 or higher. The desirability of using a standardized
HI
procedure
Newcastle
based
on
a freeze-dried
disease serum
has been pro
(1963). In addition, recommendations have been made for standardizing reagents and the method of reporting results (von Sprockhoff, 1961). The HI test depends on the presence of circulating antibody at a significant level. In Newcastle disease, this level is usually posed
by
reached
Lessing
five to ten days after
(Hofstad, 1951; Valadao,
infection
virus without developing high titres (Millen, 1960). Among recovered birds, there is considerable variation be
the virulent
HI
serum
the
tween
titres
In
(Bonaduce,
of
individuals
mild forms of the exposed to infec
HI
test
(Asplin etal, 1949).
Several authors have reported that the test using serum is more satisfactory
HI
than post mortem examination for routine diagnosis (Nitzschke and Venske, 1956; Puteanus,
1953;
Schlegel-Oprecht
and
Fey, 1953). Osteen and Anderson (1948) considered
SN
the
HI
as accurate
test
as the
test.
diagnostic purposes, the HI test found satisfactory when applied been has to organ or tissue extracts, blood clots
For
from dead birds (Mitscherlich and Gurturk,
1952; Nitzschke
Weidenmuller,
and Venske,
1956;
1955; Woernle and Sieg-
mann, 1954) or egg yolk (Bornstein et al, 1952; Schmittle and Millen, 1948; Weiden muller, 1955). Extracts of liver, spleen have been found suit
or two days after the first respiratory symptoms appear (Fabricant, 1950). However, cer
and proventriculus
tain
(1953) found there was no advantage to using organ extracts. In contrast, the HI
mammalian
component 62
which
sera
1955),
contain
a labile
also inhibits
haemag-
able for these purposes However,
(Leitner, 1954). Fey and
Schlegel-Oprecht
titres
of
egg yolks have been considered
reliable
a
of the serum
index
quantitative
of
the hen (Bornstein et al, 1952). Zargar and Pomeroy (1949) have des
titres
cribed
HI
a rapid
plate test in which one
loopful of whole blood from the wing vein was mixed with Newcastle disease antigen on a glass plate. Topolnik and Hallauer (1950) found this test gave positive results in fowls which had been infected five or more
days
using
whole
A micro-method has been useful for
blood and
diagnostic
investi
(Bamberger and Elek, 1955). rapid plate test using serum instead of whole blood has been reported by Dinter et al. (1948) and Luginbuhl and Jungherr ( 1949). The latter authors found that in 93 per cent of cases, this test gave gations
A
technique.
These
poultry.
virus,
designated
Yucaipa, which
results to the tube dilution Walker (1952) compared the
rapid plate tests, using either whole blood or serum, with the HI tube test: in 21 birds there was disagreement in one in
serologically ease virus
(Bankowski and Corstvet, 1961). Another agent capable of agglutinating chicken red blood cells is Mycoplasma gallisepticum, the causal agent of chronic respiratory disease (Fahey and Crawley, 1954; Markham and Wong, 1952).
Haemolysis The ability of the virus to haemolyse cells in vitro (Burnet, 1949, 1950; Kilham, 1949; McCollum and Brandly, 1955a) has been used by Kahnke (1951) and Nilakantan et al. (1963) in a haemolysis inhibition test for the detection of antibodies to Newcastle disease virus. These authors showed that a correlation existed between the haemolysis inhibition red blood
test and the
HI
test.
The serum titre which inhibits haemoly sis corresponds closely with the HI titre of the serum (Burnet and Lind, 1950); whereas haemolysis involve
0°C for 20 minutes before mixing with formolized antigen on a glass plate. In a
recently vaccinated
flock, this test showed
agglutinins
the
before
HI
titres became
significant. study of four diagnostic made by Schoenaers and
comparative
methods
strain
immunologically and distinct from Newcastle dis
at
A
include
is
A modified agglutination test has been described by Raggi ( 1960) in which each serum sample was centrifuged stance only.
a
agents
myxovirus
previously.
immunological
comparable
in
tions a
was
and elution appear to
and (Sagik 1957). Under suitable conditions, a single virus particle per red blood cell can cause haemolysis (Sagik and Levine, separate
reactions
Levine,
1957). Furthermore, the ability to lyse chicken red blood cells depends on the
of only one intact haemolysin particle (Wilson, 1958).
presence
Cotteleer (1956). The techniques examined were those described by Monti (1952), Mitscherlich et al. (1954), Geurden and
unit
Devos
flat plates and that approximately
mann
(1955) and Woernel and Sieg(1952). It was concluded that the
haemagglutination disease-sensitized
test using chicken
Newcastle
red blood
cells
(Bang and Libert, 1949; Geurden
and
Devos,
1955) was the most reliable.
per virus
Radiation studies on the haemolysin
in
dicate that the haemolysin
are
molecules
15 such
are arranged on the virus sur
molecules
face. Although a virus particle
with only
one
can
intact
chicken much
unit
haemolysin
red blood slower
lyse
cells, it does so at a
rate than a virus
with
15
units (Wilson, 1958). haemolytic activity in Newcastle
intact haemolysin
Other Haemagglutinating In addition to Newcastle
Agents
disease virus,
other haemagglutinating agents have been recovered from respiratory disease condi
Full disease
virus
suspensions
has
not
been
obtained without pretreatment of the virus by
techniques
thawing,
such
precipitation,
as
freezing
extensive
and
dialysis 63
(Burnet and Lind, 1950; Granoff and 1954; Vrtiak et al, 1959; Wilson, 1962b) or drying (Wilson, 1958). Such treatment of the Bl strain of the virus has resulted in the ability to agglutinate and haemolyse human red blood cells (Liu, 1952). It has also been shown that fluoroHenle,
carbon treatment may be used to unmask the Newcastle
(Wilson, pressure
disease
virus
haemolysin
1 962a) . The application of high to Newcastle disease virus has
been suggested as a means of releasing the haemolysin
(Atanasiu et al, 1955). In of 15 strains of New
an examination
of the fluorescent antibody for bone marrow cultures have been given by Jerushalmy et al. (1963). disease studies with Newcastle Other serum
and
technique
fluorescent
have been reported
antibodies
by Reda et al. (1964).
Serum Electrophoresis Using the serum electrophoresis
tech
nique, Lukacevic et al. (1958) found that fowls serum samples from vaccinated showed
in gamma
increase
a significant
globulins.
castle disease virus, all caused haemolysis
of fowl erythrocytes (Nilakantan et al., 1963). The heating of the virus in amnioallantoic fluid at 50°C for 30 minutes abolished its haemolytic
Serum or Virus Neutralization
(SN Test) associated with the
The antibodies
activity.
HI
and the SN tests do not appear to be a
Intradermal Inoculation The intradermal
inoculation of
a skin-
adapted Newcastle
disease virus has pro duced a skin reaction only in susceptible
(Yates et al, 1953). Chickens which had shown respiratory symptoms for at least one day had sufficient immunity to prevent the development of this skin chickens
reaction.
antibody has been prepared
chicks,
and this antibody detected virus antigen in chicks three hours after infection (Mae-
been positive
strone and Coffin, 1961). The antigen was identified in the larynx and vascular walls of internal organs and it was suggested that, for diagnostic
purposes, it would
be
if tracheal scrapings and impres sion smears from the brain, spleen and lungs were submitted. Maestrone and
adequate
Coffin (1961) showed also that, the virus survived
64
serum-neutralization at
time
a
glutination-inhibition strated
than
was
titres when not
have
haemagdemon
1959). SN titres have persisted much longer than HI
(Schmidt,
generally
titres (Hanson et
al,
1950), but not always
(Fabricant, 1949a). Only one antibody molecule is required for the inactivation of an infectious part icle (Rubin and Franklin, 1957). The
at room
neutralization
10 days;
embryonating
after fixation of the smears in acetone for 1 0 minutes, the survival period was one month. Details of the preparation of fluorescent antibody from guinea-pig whereas
level earlier
SN titres (Fabricant, 1949a; Osteen and Anderson, 1948) as shown in Figure 13. However, in a number of vaccinated
from the serum globulin of immune chicks
temperature,
is their mechanism
peared at a diagnostic
Fluorescent Antibody Fluorescent
neither
single entity;
1948; of reaction the same (Beach, Brandly, et al, 1947; Hanson et al, 1950; Schmittle, 1953). However, the HI and SN titres of experimentally or naturally infected fowls have shown a close correla tion (Beach, 1948; Brandly et al, 1947), especially during the ascending phase of immunity. Generally, HI titres have ap
mal chicken
indices, eggs,
as
determined
in
have shown that nor
serum does not contain more
than 11 neutralizing doses per ml. (Cun ningham, 1951; Doll et al, 1950c, d). In titration
of highly virulent
strains,
grow-
A COMPARISON OF THE RESULTS OF THE HI AND SN TESTS S N
HI
Titre
Titre
sss
I I I I I I I I I I I I I I I I I I I I I I
0
7
14
21
I
I
28
35
Days (Redrawn from Fabricant,
Figure
ing or stead
1949)
13.
adult chickens
of chick
have been used in-
embryos.
In both chicken
and embryo 1 ,000 infective
titrations,
neutralization
of
doses of virus has been ob65
1948). In hyperimmunized
chickens, there ap pears to be a direct relationship between a high HI titre and the protective value
of the serum for day-old
chicks
(Bodon
et al., 1952). However, there has been no correlation between the level of specific neutralizing and the known antibody disease history (Bankowski, 1961c; ard and Jungherr, 1944).
Min-
found very sensitive and specific for dem onstrating Newcastle disease virus anti body in chicken serum (Wolfe et al., and Rice, 1954; Nitz1949; Boulanger schke, 1956; Wenner et al, 1950). There has been good correlation between the serum titres obtained in the complement-fixation
and
test
test (Boulanger and Rice,
in
HI
the
1954; Russeff,
1956a). In addition, the complement-fixa tion test has been used to distinguish from strain of velogenic lentogenic
a
of identifying Newcastle disease (Beach, 1944; Doyle, 1935; Osteen and Anderson,
a
tained and this has been used as a means
(Galassi and Gramezi,
virus
1959).
Techniques used for neutralization tests have been described in detail in Anon.
Horsfall (1953). De-embryonated
were
eggs
found
particularly suitable by Greuel (1963b). Results have shown that the demonstrable antiviral activity or titre of yolks from eggs laid by immune
to 100-fold
lower
hens is generally 10than the serum titres
et al, 1946c). On this basis, it has been thought that eggs may be just as satisfactory as blood serum
of these hens (Brandly
for assessing the Newcastle disease status of a flock (Brandly et al, 1946c).
Complement- Fixation Tests The direct complement-fixation test has been of little value in the diagnosis of
Precipitation Test (Ouchterlony Double Diffusion Plate Technique) The application of this technique (Ouchterlony, 1948) to Newcastle disease has
been
described
and
by
Cunningham
Woernle and
Brunner (1960) (1961) . Although the reaction has been considered to be specific (Woernle and Brunner, 1961), no precipitin has been found in sera from high percentage of infected or hyperimmunized naturally fowls (Schoop and Wachendorfer, 1960). Furthermore, the degree of precipitation by
a
(1946a) ; and by Brandly et al. (1946d) and Minard and Jungherr (1944). They have also been discussed by Bang and Foard (1956a, b) and by Tyrrell and
has not corresponded to the haemagglutitre, and organ extracts from in
tination
fowls have been negative.
fected
satisfactory
More
results have been reported
by
Wachendorfer (1961), who demonstrated
Newcastle disease in chickens (Rice, 1961) as a result of incompatibility between cer tain avian antibody-antigen complexes
naturally
and
cinated with either adsorbed or "drinking-
guinea-pig
complement
(Brumfield
precipitins
by gel diffusion infected
or
in blood
He also reported that chicks
fowls.
of
hyperimmunized vac
al, 1961). Recently, Rott and Reda (1963) have reported the identification of
water" vaccine showed no precipitin reac
a soluble
have
et
antigen, termed
"nucleoprotein
antigen," which has inhibited complement. tion
test
the (i.e.,
fixation), or
66
a
indirect
complement-fixa
inhibition of complement modification of has been it,
However,
tion. Similar results in vaccinated chickens been
reported
by
Guillon
et
al.
(1963) who found that the re-infection of vaccinated chicks with field virus resulted in hyperimmunization which yielded
positive
gel diffusion
results.
DIAGNOSIS
BY VIRUS ISOLATION
Serological methods may be sufficiently to confirm field information, but new foci or extensions of the disease should be confirmed by recovery and identification of the virus (Beach, 1948; Guillon et al, 1963; Osteen and Ander 1948; Puteanus, Walker, son, 1953; 1948). After making a comparative study of four serological methods, Schoenaers and Cotteleer (1956) concluded that the inoculation of the developing chick em bryo was the best method. For example, an Indonesian strain of the virus has been propagated in the allantoic sac of develop ing chicken embryos (Martini and Koerjana, 1949). Specimens for virus isolation should be from cases in the early or even prodromal stage of the disease, and pre ferably from the younger age groups (Anon., 1946a; Beaudette et al, 1948a; Brandly et al, 1946d). accurate
brain (Mitscherlich et al, 1954). Brain tissue probably has the greatest concentra tion
(Mantovani,
of virus
1948).
How
ever, Kohler
(1960) observed consider able amounts of Newcastle disease virus in the leucocytes for three days after arti ficial infection. The Hertfordshire vaccine virus has been recovered from the central nervous
system 6 to 14 days after vac
cination
(Salyi and Hodosy, 1952). The
virus
not
as
consistently
bone marrow
as
in respiratory
is
tissue (Baskaya
et
al,
present
in
and spleen
1952).
The rate of multiplication of an avirulent Newcastle
disease virus
tissues following intramuscular
in different inoculation
of susceptible 10-week-old chickens was studied by Karzon and Bang (1951a) who found the decline in virus titre was slowest in brain tissue (Figure 14). It that samples from follows, therefore, pooled tissues give a higher re rate than samples from any one tissue (Beaudette et al, 1948a; Pannu and several
Distribution of Virus in the Body Asdell and Hanson
( 1 960)
covery
Bankowski, 1962).
and Hofstad
(1951) showed that the virus, when in into a susceptible fowl, spread rapidly from the site of inoculation and could be detected in almost all tissues within 48 hours, and in all tissues within 72 hours. Electron micrograph studies of red blood cells have shown the maximum number of virus-like particles to be pre sent 96 hours after inoculation, at which time the infected fowls have shown symptoms of Newcastle disease (Reagan
Embryonating Eggs
jected
et
al,
1954c). results indicate
Nevertheless, that virus
laboratory
may be more
from some tissues than from others. The following tissues have been found particularly suitable: lung and trachea (Hofstad, 1951; Miller and Miller, 1950), respiratory and spleen readily
recovered
(Baskaya
et
al,
1948a; Beaudette
1952; Beaudette and Black,
et
1946)
al, and
Although embryonating used,
generally
successful
hens' eggs are results have
duck obtained with embryonating (Collier and Dinger, 1950) or young ducklings (Komaroff and Goldschmitt,
been eggs
1946a).
Preparation of Inoculum In some early work, unfiltered
tissue
were not centrifuged and no was added (Beach, 1943; Beau
suspensions antibiotic dette and
Suspensions
Black,
1946; Walker,
were filtered
through
1948). Berke-
Mandler and Seitz filters (Brandly 1940; 1946d; Iyer and Dobson, Rodier, 1928). Among the antibiotics used to inhibit bacterial contamination, the relative values of penicillin alone feld, et
al,
67
TISSUE DISTRIBUTION OF NEWCASTLE DISEASE VIRUS AFTER INTRAMUSCULAR VACCINATION OF 10-WEEK-OLD CHICKENS Log of
LDM
Virus
\
5-
0
Spleen
— ■ Lun9
5
3
1
7
9
Days after Inoculation (Redrawn from Karzon and Bang, 1951)
Figure 14.
(Brandly 1950),
et
al,
1946d;
streptomycin
Cordier
alone
et
al,
(Delaplane,
'1947; Thompson and Osteen, 1948), or a mixture
of penicillin and streptomycin
(Beaudette et al, 1948b) Later studies showed was capable
of inactivating
that
penicillin
the virus,
to
of penicillin and the duration of incubation of the mixture before inocula tion of the embryonating (Kohn, eggs 1953). In a comparison of filtration and antibiotic treatment (penicillin and strep tomycin), it was found that in 22 samples 68
treated material
etal,
was positive
(Beaudette
1949).
were reported.
an extent that depended on the concentra tion
the filtrate was negative for virus on egg inoculation, whereas the antibiotic-
Route of Inoculation Inoculation into the allantoic
sac has
(Beaudette et al, 1948b; Thompson and Osteen, 1948), and
been commonly used various
techniques
have
been described
(Anon; 1946a; Beaudette et al, 1952; Mc Carthy and Dumbell, 1961). A combina tion of inoculation of the chorio-allantoic
and of the allantoic sac, through the same hole, has been found of in the primary isolation valuable viruses (Fabricant, 1957; Gorham, 1957). This technique has also reduced embryo mortality due to trauma (Bueno et al.,
virus isolated from chickens,
1961).
absence
membrane
Temperature of Egg Incubation Sinha (1958) has reported that inocul ated embryos
have died
9 to
hours
15
earlier when incubated at 37°C instead of 35°C. Bang (1948) found that good virus growth occurred at temperatures from 35°
to 41 °C, and von Sprockhoff (1960)
recorded no difference in gross lesions and
HA
titres between embryos
incubated
at
35.5° and 38°C. In contrast, Zuschek et al.
(1959) have reported that in embryonatactivity followed a log linear response to temperature.
ing eggs haemagglutinin
Blind Passages Beaudette
Black
and
that approximately
(1946) found of a series
two-thirds
of 239 negative samples were determined as negative on the first inoculation of embryonating 19 were
eggs.
Of
26 positive
as such on the first
determined
egg passage; the remaining
killed
samples,
seven samples
the second passage. Thus, it was concluded that blind every
in
embryo
passages were unnecessary in the diagnosis of Newcastle
disease (Beaudette,
1948a). content of
In contrast, where the virus the inoculum is extremely low, some degree of virus adaptation to the embryonating egg may be necessary (Brandly et al, 1946d; Anon., 1959).
Embryonic Mortality Suspensions
of
trachea,
spleen have produced in 12-day
embryos
lung
and
death in 72 hours
(Perez and Gonzalez,
1951). Using 9- to 10-day embryonating chicken eggs, Vrtiak and Polony (1962) examined
32 strains
of Newcastle
disease
ducks and a
Generally, all strains killed the embryos within 24 to 48 hours; occasion ally an embryo survived 72 hours. How pheasant.
ever, several factors
influence
embryonic
or of parental antibody (Brandly et al., 1946c), the virulence of the virus (Anon., 1959; Beach, 1943; Chu, 1958; Hitchner et al., 1951a; Minard and Jungherr, 1944), the temperature of incuba tion, and the route of inoculation. Death of the embryo has occurred somewhat these include
mortality;
earlier
after intravenous
the presence
than after allan
inoculation (Hanson et al., 1947). However, Bang (1948) found that neither the temperature of incubation nor the route of inoculation of embryos had a consistent effect on the measurement of virus activity. For the latter purpose, the toic
50 per cent embryo
mortality
was used.
Either a single particle of virulent New castle disease virus in the inoculum or suc cessful attack of a single locus has been considered sufficient to kill the 10-day-old embryo
(Nadel
et el.,
1957).
Lesions in Embryos In embryos, the virus has been found to the cells of destructive and proliferating rapidly differentiating tissue (Williamson et al. , 1 956). The results of inoculating chick embryos by the chorio specifically
allantoic, methods
amniotic
and
sub-allantoic
have been described
by Burnet
(1942). Macroscopic Lesions. These vary with the strain of infecting virus. With the more virulent (velogenic) strains, small or large are common, especially on the wings (Iyer, 1943) and legs (Iyer and Dobson, 1 940) , and over the cranium and dorsal body surfaces (Jungherr et al., haemorrhages
1946). The yolk sac is usually congested. With the lentogenic strains, haemorr hages are seldom seen; but stunting and
curling of embryos dying on the fourth and succeeding
days have been observed 69
(Hitchner et al., 1951; Anon., 1959). The curling of embryos into a ball-like form has been considered pathognomonic (Fabricant, bronchitis infectious of 1949b).
Microscopic Lesions.
The chorio
has shown prolifera
membrane
allantoic
toic
membrane
in the chicken embryo has been measured of by an increase in the concentration fluid proteins in the infected allantoic and
(Kilbourne
tion of the ectoderm with vacuolation (Iyer and Dobson, 1940), increased density of
of the cells et al., Kilham 1934; Ferry, and (Burnet and necrosis
the cytoplasm,
(Burnet, 1942) and in the allan (Granoff, 1955). The disease virus Newcastle development of
embryos
1951). Other changes have been difficult to lesions from non-specific differentiate (Jungherr et al, 1946). Thus, in an experi
Liu and Bang (1953), virulent strain of virus did not produce significant changes in any organ of em bryos up to the time of death. The mesoderm has shown haemorrhages and oedema (Burnet, 1936). In addition, defective development of the lens, otocyst or caudal portion of the neural tube has
ment conducted by
This 1949). concentration has been Horsfall,
increased protein attributed to the reaction
of the develop
ing embryo. After examination of electron micrographs, Mussgay and Weibel (1962) that morphologically intact concluded particles
of Newcastle
capable of entering
disease virus were
cells of the develop
ing chicken embryo.
a
been described
(Williamson
and
inclusion
of
disintegration
cytoplasmic
cells in tracheal
al,
1956). lesions include vacuola
Other microscopic tion
et
smears
(Burnet,
1942);
bodies in cells of allantoic
mem
liver smears (Collier
brane and embryonic
and Dinger, 1950) ; and multiple capillary haemorrhages in the spinal cord (Jung herr et
al,
1946).
disease virus has an ability
to spread throughout bryonating centration
Greuel (1959) found onated eggs and
more
that de-embry-
be used more rapidly efficiently than embryonated could
The virus was demonstrated in 37 infected fowls out of 45 experimentally by the inoculation of brain suspension on to the chorio-allantoic membrane of deeggs.
embryonating
eggs.
In contrast,
only
29
tissues were positive by the inoculation of chicken embryos. De-embryonated eggs permitted the use of a larger inoculum and were not affected by factors which kill
(Greuel, 1963b). In embryos comparative studies with de-embryonated eggs in which the chorio-allantoic mem brane has been either removed or left virus intact, virtually all the inoculated chicken
Distribution of Virus in Embryos Newcastle
De-Embryonated Eggs
7~
the developing
em-
1948). Virus con and haemagglutinating activity egg (Bang,
has been adsorbed to the chorio-allantoic cells (Nadel and Eisenstark, 1956).
have been found to vary in different fluids and
tissues
(Burnet,
of infected
chick embryos
1942; Hanson et
al,
1947),
as
in Figure 15. These differences have been associated with strain of virus
illustrated
and route of inoculation, the inoculum has not titre
of the virus
but the size of affected the final
in the allantoic
fluid
(Bang, 1948). Evidence of very active virus multipli cation has been obtained in the viscera of 70
Tissue Cultures Growth of the virus has occurred in a medium consisting of chick embryo tissue and plasma (Topacio, 1934), and in cul tures of cells from a variety of animal Brandt, species (Bankowski et al, 1960; 1961; Das and Goldberg, 1961; Fontanelli et al, 1960; Franklin et al, 1957; Gelenczei and Bordt, 1960; Mason and Kauf-
SELECTIVE INFECTION BY NEWCASTLE DISEASE VIRUS Haemagglutination
Titre Allantoic Fluid Amnionic
Membrane
Amnionic
Fluid
Hours Post Inoculation (Redrawn from Hanson ef al, 1947)
Figure 15.
man, ef
al,
1961b; 1960).
Rubin
al,
1957; Vrtiak
cells in plasma media or roller tubes, re
Studies with chick embryo
ported by Gey and Bang (1951) and Bang
et
71
Lynn and
( 1 953a) , have been reviewed by
Morgan (1954). Growth in tissue culture has been used as a means of virus identification because effect has been shown to be
the cytotoxic
caused by the viral particle
(Prince and
Treatment of 1957). of Newcastle disease virus
Ginsberg,
field
strains
with
nitrous
has
acid
characterized
resulted
by different
in
mutations
plaque appear (Granoff, 1961;
ances in tissue culture Thiry, 1963). It is considered that the kill ing of mammalian tumour cells by New
castle disease virus is not solely a surface reaction
and
(Prince
Serum neutralization formed
using
magglutinin endpoints
Ginsberg,
tests
1957). have been per or
cytopathogenicity
hae-
formation for determining (Crowther, 1963; Goldwasser
eggs for the titration and isola of the virus (Fastier, 1954; Gold wasser and Kohn, 1957; Matewa, 1960). In addition, tissue cultures have been a a mildly patho means of distinguishing genic from a more pathogenic strain of virus (Mussgay, 1960). The difference in the degree of cytopathic effect between with their patho strains has coincided and genicity for chickens (Subramanyam Pomeroy, 1 960) . Thus a virulent strain of
bryonating tion
Newcastle
disease
has
virus
destroyed
in tissue culture more rapidly than an avirulent strain (Bang and War wick, 1957). From the economic point of view, Jakubik (1962) considered the fibroblasts
tissue culture
method
to be 85 per cent
cheaper and to involve about half the work required for isolation in the chick embryo.
and Kohn, 1957; Levine and Sagik, 1956;
Mason and Kaufman, 1955; Pigoury et 1962; Rubin and Franklin, 1957; Seif-
Mixed Virus Infections
al,
fert,
1955; Subramanyam
strain
Considerable
and Pomeroy,
variation
1960).
resistance of Newcastle
According to Seadale and Winterfield (1956), tissue culture titrations based on
avian viruses to inactivation
cytopathogenicity
are somewhat more sen
sitive than virus assays in chick embryos.
The virus has been titrated in tissue cul to show colour using indicators change resulting from the metabolism of tures
cells
non-infected
(Durand
and
Eisen-
been inactivated by hydroxylamine, as
an
attenuated
strain
has given titres approximate
1 log. less than embryonated
egg titres
expressed as LD.n units per ml. (Bower, 1958). A close correlation has been found in in vitro titration glutinating
between the haemag-
and haemadsorbing
of the virus
properties
(Rossi, 1961a). Tissue cul tures have been found comparable to em-
72
where
(Rott
and
1962). Moreover, it has only been possible to resolve certain mixtures
of Newcastle
by physical-chemical
ly
has
Schafer,
immune serum. et al., 1957)
by physical-
means has been found (Quiroz and Hanson, 1958). A highly pathogenic strain of Newcastle disease virus has not
infectious
Titration by the plaque method (Rubin
the
chemical
1959). This method has also been found suitable for titrating the specific
stark,
in
disease and other
disease,
bronchitis
laryngotracheitis,
and fowl pox viruses
procedures (Quiroz Hanson, 1958). Brandly et al. (1946d); Hanson (1954) and Thompson
and
(1954) showed
sera that hyperimmune neutralize the virus satisfactorily of Newcastle disease, fowl plague and
would
bronchitis and a chronic res piratory disease agent in dual infections so that the presence of each virus could infectious
be demonstrated
in embryonating
eggs.
BY INOCULATION
DIAGNOSIS Chickens
Pigeons
The use of susceptible chickens, free passive antibody, for the isolation and identification of Newcastle disease virus has been valuable under certain cir cumstances (Anon., 1959b; Beach, 1943; Brandly et al., 1946d; Gordon and Asplin, 1947; Walker, 1948). The use of suscep tible chickens was regarded by Grausgrufrom
ber (1958)
as being time-consuming
and
Furthermore, due to the varia tion in the susceptibility of individual fowls and differences in the virulence of field strains of virus (Vrtiak and Polony, 1962), Beller and Siegmann (1955) con
expensive.
cluded
that diagnosis
by the inoculation
of fowls was unsatisfactory should
be
supplemented
by itself and by
serological
of fowl has facilitated the isolation of virus (Scott et al., 1956). Karzon and methods. Nevertheless,
the inoculation
a healthy
Bang
(1951a)
showed
that
whereas
a
velogenic and a mesogenic strain increased at the same rate in the extra-neural the
velogenic
growth rate in the brain.
Figure
tissues,
strain caused an increased
In
susceptible pigeons, symptoms usual
ly appear six days after inoculation (Figure 16) and death occurs a few days later still (Doyle, 1935). The features of the experimental
disease in pigeons have
been described by Dobson (1939), Doyle
(1935) and Kuppuswamy (1955). The of the pigeon was shown by Kaschula (1951) to influence the de velopment of symptoms. Pigeons inocul
site of inoculation
ated in the neck
showed
a more
rapid
course and developed paralysis of the neck
quicker than those inoculated in the wings and legs.
Olah and Palatka (1962) made a com study of the pathogenicity of several strains of Newcastle disease virus inoculation of pigeons. by intracerebral prehensive
An
intracerebral
pathogenicity
index was
used, giving death a value of 4, illness 2 and no ill effect zero.
A virulent
virus gave
an index of 3; a strain designated Lederle was 2; the Roakin strain was 0.9; and Bl,
F
and LaSota each gave an index of zero (Olah and Palatka, 1963).
16. — Newcastle disease in the pigeon. (Crown Copyright. Reproduced by permission of the Controller of Her Malesty's Stationery Office, London.) 73
Ducks When compared with chickens, other domestic poultry are more resistant to artificial infection with the virus. How ever, ducks
have been used by Schofield
and Hutsen
(1952) in the identification
of the disease.
Laboratory
are
(Brandly
al,
et
generally
al,
et
in-
1946d;
Groupe
1951;
and
evidence of a previous
New
castle disease infection may be obtained suitable live specimens with
by inoculating
known dose
1947).
of fully
test
virulent
Gordon
1959;
This
and
Newcastle name
drewes
An
virus
Asplin,
should be controlled
by
disease virus has been given
Myxovirus
et al.,
multiforme (An-
1955;
Waterson,
early name for the virus was Tortor
by Brandley et al. ( 1946d) , Wilner (1964), and Andrewes and Worthington (1959). marized
and
Chow
(1958)
these methods
have
sum
and criteria
and
have used them to identify a field isolate.
Various criteria castle disease virus
for identifying
New
have been reported
(1956). They have been summarized by Ozawa and Chow (1958) as follows: — Pathogenicity for embryonating by Hanson
— — 74
bral
with
routes
several
of young
inoculation
et al.
chicken eggs.
Mean death time
for minimum
lethal dose. Pathogenicity
ment young
of the central mice
hamsters
etal,
the
nervous
have generally
lethal for chickens
has
system been
in
highly
(Carlotto, 1954; Upton
1953a).
EXPOSURE
inoculation
of
fully
susceptible
chickens.
The value of the challenge method of was demonstrated by Bankowski (1961c) who found some chickens with a negative HI titre refractory to a challenge dose of virus. diagnosis
for day-old
—
chick
ens (neuropathic Pathogenicity
index). for day-old
(IN)
ens by intranasal
1962).
furens (Barger et al, 1958). Criteria for establishing its identity have been given
Ozawa
by
Newcastle disease virus; and the intracere
OF NEWCASTLE DISEASE VIRUS
IDENTIFICATION the
infected
be
BY CHALLENGE
DIAGNOSIS
(Brandly,
can
Bang,
et el, 1956). Strains which have induced severe involve
mammals
tracerebrally
a
that kittens and adult cats
1952;
a human infection (Reagan
resistant to the virus unless inoculated
Definitive
have reported
1956; Kilham et
and
been used for the recovery of virus from
Laboratory Mammals
Brueckner
Nevertheless,
al, 1952; Liu Upton et al, 1955). Luttrell and Bang (1958)
Dougherty,
muscular
(IM)
inoculations.
—
Pathogenicity
—
(IN, IM). Pathogenicity for chickens (IN, IM).
—
chick
and intra
for 3-day-old
chick
ens
5-week-old
Disease manifestation.
— —
Antigenicity (protective
antibody).
Serum
(homolog
—
ous LDS0 neutralized). with Haemagglutination
neutralization
chicken
and other animal erythrocytes.
— —
Haemagglutination
—
Heat stability of embryo infectivity
—
inhibition tests. Heat stability of haemagglutination at 56°C. at 56°C.
Mouse index).
toxicity
(neuropathic
DIFFERENTIAL The great
of symptoms
variety
lesions exhibited
in Newcastle
outlined in the preceding
been
and
disease has section.
form
The occurrence of an asymptomatic has
been
discussed
on
page
57. These
of the disease make differential
features diagnosis
difficult
laboratory
procedures
unless is
recourse
to
New
adopted.
castle disease virus is classified as a mem myxovirus group. The term myxovirus" includes fowl plague virus, virus N, duck influenza virus, tern virus, myxovirus Yucaipa and perhaps
DIAGNOSIS ways, was a separate entity caused by a virus immunologically distinct from the virus of fowl plague. Although this distinc tion between the two viruses was not at first recognized
by some authors (Hutra 1938; Leynen, 1935; Manninger,
et al.,
1936; Picard, 1934),
1932, firmed
other
workers
ber of the
1934;
"avian
Thus, Ivanova
others.
ceptible to inoculation
In the following pages, the similarities between Newcastle
summarized.
but it may serve as a
early investigators
has been discussed by
Hutra et al. (1938); Jacotot (1950); Jacotot and Vallee (1949); Lesbouyries (1941, 1951); and Traub (1942). In Europe and some other areas where fowl plague (geflugelpest) was recognized before 1926, the later appearance of the
form of Newcastle
often not immediately instances,
disease was
identified.
laboratory
that a clinical diagnosis disease
Thus, in
results
have
of fowl fowl
resembling
plague, was made in error (Bakos and Nordberg, 1949; Berke and Golem, 1949; Jacotot and Vallée, 1949). In the original identification of New castle disease virus (Doyle, 1927, 1935), it was
shown
virus
the ery
agglutinated
with a more stable antigen-antibody
union
than with Newcastle
(Hall-
disease virus
(1960) has concluded that the of Newcastle disease virus into classical fowl plague virus is a pos is that Another consideration sibility. Schmidt
The difficulty of distinguishing between disease and fowl pest (now generally called fowl plague) that faced
a
fowl plague
transformation
Newcastle
or
with three different
strains of fowl plague virus. Another dif
auer and Kronauer, 1954; Kunst, 1949). In more recent studies of this problem,
Fowl Plague
plague,
1950).
throcytes of a larger number of species and
Viral Diseases
shown
Schafer,
(1963) showed that chickens immunized with vaccine strains of Newcastle disease virus were fully sus
ference between the two viruses was that
means complete,
some
1931;
et al.
disease
guide.
classical
Purchase,
of other avian diseases are This summary is not by any
and differences
and a number
it was con
(Burnet and Ferry, 1934; Lush, 1943; Nakamura et al., by
that Newcastle
disease, al
though it resembled fowl plague in many
Newcastle
disease virus
may be identical
to atypical fowl plague virus (Beller, 1953; Schafer, 1950). There is, therefore, constant need to dif or antigenically
related
ferentiate between the two diseases, especi ally in countries where both might co exist (Lucam, 1949c; Vittoz, 1938, 1963). Such
differentiation
has been
made
using fowl plague and Newcastle
by
disease
(Nechvatal, 1950), and by cross immunity tests (Kujumgiev, 1950). In vaccines other
instances,
between
attempts to differentiate disease and fowl
Newcastle
plague have been made without
isolation
In addition
of the virus (Anon., to the serological
1948). and virus isolation
so far discussed
(pages
following distinguish
methods between
have
tests
59 to 74), the been
Newcastle
used
to
disease
and fowl plague: 75
TABLE
9 —
Characteristics of Classical Fowl Plague
and Typical Newcastle
Disease Compared
Fowl plague Incubation period Duration of disease Symptoms
Lesions
Infectivity of blood Virus haemagglutination
Newcastle disease
Average 1 54-214 days. A few hours or longer. Often none. Malaise, diarrhoea, oedema of head and append
Average 4-6 days. Three days or longer. Respiratory symptoms, diar rhoea, nervous disturbances
ages.
in survivors. May be none. Haemorrhages
May be none. Haemorrhages on heart and in proventriculus and intestines. Sero-gelatinous exu date in lungs, pericardium and subcutis. Virus present in high dilutions of blood. With erythrocytes of many spe cies. Slow elution of virus.
Virus haemolysin1 Lesions in inoculated embryos2
often involving lymphoid patches of intestine.
Virus present in lower dilu tions. Quicker elution of virus.
Negative.
Present.
Cytoplasmic inclusions absent. Haemorrhages in skeletal mus
Cytoplasmic inclusions. Petechiae on body surface.
cles. ' Andrewes and Worthington 2 Burnet and Ferry (1934)
(1959)
Pigeons. During the years immediately following the first identification of New castle disease, the virus was differentiated
from that of fowl plague principally by pigeons, which are susceptible by inoculation to Newcastle disease but re sistant to fowl plague (Dobson, 1952; Doyle, 1933; Purchase, 1931). Additional using
references are given on page 73.
Mice. Although this animal used, results indicate
is seldom
that mice are gen
erally more susceptible to the virus of fowl plague than to that of Newcastle
disease
(Kranevald and Nasoetion, 1941; Kunst, 1949). However, little difference has been reported
by Andrewes
and Worthington
examined
by
Demnity
and
etal,
1948).
Clinical symptoms and post mortem findings. The major differences between the typical forms of Newcastle disease and fowl plague (summarized in Table 9) have been described by Beaudette ( 1951c), Fritzsche and Gerriets (1962), Jezierski
(1959).
(1953), Jungherr
The agglutination of rabbit erythrocytes by fowl plague virus
(1946, 1952).
but not by Newcastle
Avian Encephalomyelitis
Haemagglutination.
disease virus
been used as a differential
Aziz the
et
HI
guish 76
has
(Abdel al, 1960) . The possibility of using test with high litre sera to distin
between
the
two
test
viruses
was
Schneider
(1950). Physical properties. The "Herts" strain of Newcastle disease virus has been found to have a pH stability of 5.5 to 7.5 and a mean size by electron microscopy of 150 ±50 microns; whereas the corresponding figures for fowl plague virus have been pH 7.0 to 9.0 and 83 ±15 microns (Elford
et al.
The comparative encephalomyelitis
(1946) and Stubbs
pathology
and Newcastle
of avian disease
has been reported by Jungherr and Minard
the (1944) . In avian encephalomyelitis, central nervous system has shown exten
perivascular
sive
whereas has
the
foci
granulomatous nervous
peripheral
system
shown some myelin degeneration.
this disease,
tom and
there is no respiratory
no inflammation
tory tract.
In
caused
the developing lesion
no
in
symp
of the respira chicken
avian encephalomyelitis
bryo,
In
virus
visceral
em has
organs
(Casorso and Jungherr, 1959).
Avian Leucosis Complex The neuralymphomatosis disease (Biggs,
Newcastle
Newcastle
Mohr
has been applied
castle
disease
gel
disease
has
been
diffusion
technique
to the diagnosis of New
and
from
disease has been described
(1953). The main difference
been the presence
of lymphocytic
in Newcastle
by has
disease
in the lungs
infiltrations
cellular reactions in the
and characteristic
Duck Plague
outlined by Beaudette (1951c). Delaplane (1945) , Fabricant (1949b, 1950) and Stubbs (1946). the Recently,
1962); and the differentia
of this form of the complex
tion
The difference between infectious bron and
of
central nervous system.
Infectious Bronchitis chitis
component
the complex has been classified as Marek's
infectious
bronchitis
(Guillon et al., 1962). A diagnostic pro cedure utilizing this technique has been based on the detection of an increase in titre between two precipitin bleedings (Witter, 1962). Infectious bronchitis virus,
successive in contra
to the virus of Newcastle disease, has failed to agglutinate sperma tozoa (Chu, 1953). distinction
The first outbreak
of this disease was
reported in 1923 in The Netherlands, since then a small has
occurred
1961). petechial
in
number that
(Jansen,
country
Typical lesions haemorrhages
and
of outbreaks
include
multiple the
throughout
body. The causal virus has been shown to
immunologically
be
from
distinct
the
viruses of fowl plague and Newcastle ease
(Jansen,
dis
and Kunst,
1951; Jansen
1949).
Virus N This virus has been isolated from dis eased fowls and,
in
some
the
respects,
virus has resembled Newcastle
disease and
fowl plague viruses. Although no serolog
Infectious Laryngotracheitis Factors that enter into the differential of this disease have been re ported by Beaudette (1951c), Delaplane (1945), Stubbs (1946), and Woernle and Brunner (1961). The use of the agar gel diffusion tech nique in the diagnosis of infectious laryn gotracheitis has been reported by Jordan and Chubb (1962). Precipitin antigens diagnosis
have
been
demonstrated
of virus recovery
in the absence
by egg inoculation.
N
group
(Anon., 1946c).
and
Tern Virus Chickens virus
artificially infected with this
have shown
symptoms
after three
days and have died a few days later.
necrosis
described
N
between virus
(Rott and Schafer, 1960).
Newcastle
been
between
disease virus, some
fowl plague virus has been demonstrated (Dinter, 1949; Dinter and Bakos, 1950). Virus N has been placed in the myxovirus
though
have
and Newcastle
antigenic relationship
Under field conditions in Italy, simul taneous outbreaks of laryngotracheitis and disease
has been found
ical relationship virus
macroscopic
slight, histological cases.
have
changes
have
Al been
lesions of focal tissue
been
extensive
However, the pathological
in
severe pattern 77
from
to be distinct
has been considered
disease or fowl plague
that of Newcastle
have also been demonstrated infected turkeys
(Gale
(Urs and Becker, 1963).
1959). In a natural
outbreak
Turkey Meningo- Encephalitis
ornithosis
The first report of this disease was made by Komarov and Kalmar (1960) who described a disease of turkeys character ized by progressive paralysis of legs and wings
and
phocytic
a severe
non-purulent
lym
The causal
meningo-encephalitis.
virus was classified by Porterfield as a member
(1961) of Group B of the arthropod-
Typically, the disease caused by Myco plasma gallisepticum period
of
5 to 21
prolonged
aggravated
by exposure
viruses
(Bankowski,
to avian respiratory 1 961b) .
A pleuropneumonia-like
with neurotropic
Mild re
caused by this or other
spiratory infections agents are often
has a long incubation days and runs a more
(Chu, 1958).
course
characteristics
organism
of
Adler, 1957.)
Avian Pasteurellosis and turkeys, with a disfunc of the central nervous system and symptoms similar to those of Newcastle Chickens
tion
disease, have been found
infected
with a
Pasteurella organism localized in the brain (Fenstermacher
Ornithosis
etal,
1946).
lesion of orni
a common
thosis is an inflammation
of the air
peritoneum and pericardium.
78
chick
to those of avian encephalomyelitis
(Storz
nervous
etal., 1963).
Nutritional Deficiencies Nutritional deficiencies which can symptoms and lesions resembling
cause
those of
disease include:
— Fatty acid al, 1963).
(nutritional
.
deficiency
(Hopkins
et
Toxic Drugs and Plants Toxic substances which can cause symp toms and lesions similar to those of New castle disease include:
— Agricultural Paver,
pesticides
1961;
Gaafar
(Barden and and Turk,
1957).
— Alkyl
organophosphorus
com
(Davies et al, 1960; Lan caster etal., 1960). — Nitrofurazone (Brion and Fontaine, 1958; Klimes and Kruza, 1962). — Toxic plant seeds (Kelly et al., 1961; Placidi, 1954c). pounds
Avian Respiratory Diseases The differential diagnosis of respiratory diseases of poultry (Table 10) has been summarized by Abrams (1961 ), Caporale
(Psittacosis)
In the turkey,
an
similar
has been
of a turkey with (Cordy and encephalitis
isolated from the brain symptoms
in chickens,
symptoms
ens exhibiting
encephalomalacia)
Chronic Respiratory Disease
1960; Page,
agent of relatively low virulence
— Riboflavin deficiency. — Vitamin E deficiency
Bacterial Diseases
in artifically
has been recovered from 2-day-old
Newcastle
borne viruses.
al,
et
sacs,
These lesions
(1961), (1952), 1961),
(1945), Durrell Delaplane Chu (1958), Gordon (1956, (1953), McMartin Jungherr
(1963) and Schyns (1961).
TABLE
Disease
10 — Avian Respiratory
Diseases — Some Characteristics
(Modified from Gordon, Incubation
Duration
Spread
period
Mycoplasma gallisepticum
10-21 days
Coli
10-21 days
many
slow
weeks
14-21 days
Profuse oculona
months
slow
difficult respiration Difficult respiration, loss of weight, rejected at market Profuse nasal dis charge, difficult respiration, gasp
Infectious bronchitis
1-3 days
Infectious laryngotracheitis Fowl pox
4-21 days
Haemophilus gallinarum Aspergillus fumigatus
slow
Clinical symptoms
sal discharge, swollen face,
septicaemia
Mixed infections 'C.R.D.,
months
1963)
brief
rapid
7-1 4 days
1- 2
variable
3-4
slow
weeks days
2—4 days
2-3
rapid
weeks weeks
rapid
features
Purulent exudate sinuses, may extend to lower respiratory
tract
Pericarditis and airsacculitis
Purulent exudation, pericarditis,
ing, poor growth, etc.
airsacculitis
Sneezing, gasping,
None, or some mucus in bronchi and air sacs Blood or diphther
depressed
weeks to months
Pathological
egg
production Gasping, cough ing, blood clots Lesions on comb, wattles, face, mouth Slight oculonasal discharge Gasping
itic material trachea
in
As for symptoms Pus in sinuses Nodules in lungs
79
PART III: Control Measures CONTROL BY SLAUGHTER Effectiveness in Various Countries
In many countries in which Newcastle disease
has a variable
importation
The concern shown by national disease control agencies over the control of New
where Newcastle
castle disease is reflected by the fact that
present, there is complete
in 1962 the disease was notifiable
in 84
countries
(Table 2). In some countries where slaughter mea they have been
sures have been applied successful;
eminently
in others they have
failed to prevent the disease from becom ing established. The variety of ways in
which the disease can spread, the actual numbers of individual poultry involved, and
prevailing have
practices
trade
and
appreciably
Newcastle
free
periods.
In
been
of the disease for extended for instance, although
Canada,
since
1950 in
of British Columbia, no known outbreaks occurred in several prov inces, including Saskatchewan, Manitoba and those on the Atlantic coast, from 1 957 to 1963. Similarly, in Scotland, some coun ties, including West Lothian, Perth and Midlothian, were free from Newcastle disease from 1951 to 1961 (Anon., 1962b) districts
although
there were
outbreaks
in other
parts of the country.
In 1962, a number of countries, including Czechoslovakia, Brazil, Bulgaria, Peru and South Africa, reported
that Newcastle
fined to certain regions 1962). 80
and hatching eggs. these restrictions,
To
reduce the extent of
it has been suggested
measures be applied
basis of "infected
on the
region of a country" in
stead of "infected country" ( Vittoz, 1 964).
Southeast Ireland In County Kilkenny, southeast Ireland,
from certain coun
the disease has persisted some
countries
prohibition on of live and dead poultry
the importation
that control
In
disease does not exist at
in January 1950, two outbreaks of New castle disease were reported on the same
tries and, in others, large regions have been kept
controlled.
handicapped
disease has, however,
completely
is strictly
management
disease eradication. eradicated
ducts
the
distribution,
of poultry and poultry pro
disease was con
(FAO-WHO-OIE,
premises (Anon., 195152). Immediately, orders for the restric tion of movement and for slaughter with under the were invoked compensation Diseases of Animals Act. During subse day on different
quent field investigations,
special emphasis
was placed on tracing the chain of infec tion to and from infected flocks.
In
addi
tion, the activities of poultry dealers were closely investigated. was confirmed
Although the disease
on only 14 premises,
1,000 premises were visited
over
and tens of
of poultry were examined. Within two weeks of the first outbreaks,
thousands
it was apparent that control measures were meeting with success; and during the next month
the size of the infected
progressively
reduced.
At
the
area was end
of
March 1950, all restrictions were removed except that restocking of infected premises was not permitted until four to six weeks
after cleaning and disinfection
had been
carried
the Fowl
out under
Pest Order,
completed.
and
In this outbreak in southeast Ireland, 71 flocks, involving 1,235 poultry and 40,202 day-old chicks, were slaughtered.
adopted to control the movement of poul
The whole
state was not considered free from the disease until June 1931 (John stone, 1933).
The second outbreaks in Victoria, Aus tralia, started in October 1932 and lasted 1933. Restrictions
ment of poultry, farms
quarantine
and controlled and
try, hatching
eggs
were
At
and carcasses.
measures were taken
to restrict
first,
the im
the
on move
of infected
slaughter
outbreak
was
were en
quickly
stopped. While the outbreak lasted, certain auction rooms were reserved for the sale
of poultry from outside the quarantine area. Other facilities were designated for the slaughter and evisceration of healthy poultry from within the quarantine area (Johnstone, 1933).
infected
carcasses. Later,
of imported
the zoning
In Australia, the first outbreak occurred near the town of Wonthaggi in the state of Victoria in November 1930 (Johnstone, 1931; Albiston and Gorrie, 1942). The disease spread from the original infected area, and during the following two months it was identified on 3 1 other farms within a radius of 20 miles of Melbourne. In all, 72 farms eventually became infected and 22,284 head of poultry were involved.
forced,
measures
ancillary
portation of poultry and hatching eggs and to ensure the boiling of waste food and
Australia
until March
certain
area
restrictions
were
imposed
of poultry premises and vehicles was required. The general procedure adopted to deal with the out breaks has been described by Reid ( 1961 ) . Failure to report outbreaks was thought to be handicapping the slaughter policy (Andrews, 1948), and by the end of 1947, and the disinfection
2,222
outbreaks
had
been
confirmed
(Figure 17). In 1947, both the peracute (velogenic) and the subacute (mesogenic) forms of the disease were recognized. The control measures adopted were successful in eradi cating the peracute type by 1953, but the
of the disease could not be from many areas. Scotland re mained free from the disease from 1952
less acute forms
eradicated to 1957.
Another series of events which reveal of epidemiological aspects of the disease is associated with its eradication from the county of Lancashire (Figure a number
17).
were
Here, begun
epidemic
concentrated in
1956;
control
and by
efforts
1958 the
in one area of the county
had
For over two years there was no recrudescence of the disease in the area. This success was achieved only by full and rigorous enforcement of the slaughter policy (Ritchie, 1962). Whether this success in one section of the county of Lancashire would have been possible in the face of widespread outbreaks in other parts of the country is debatable, In July 1960, the Departmental Com mittee on Fowl Pest was appointed by the British Minister of Agriculture, Fisheries and Food. Their report (Anon., 1962b) in March was presented to Parliament concluded that 1962. The Committee
been terminated.
Great Britain The control of Newcastle disease by in Great Britain has been de in a number scribed of publications (Andrews, 1948; Anon., 1962b; Asplin et al., 1949; Callender, 1958; Dobson, 1949; Gordon et al, 1948; Gordon, 1961; Gordon and Asplin, 1947; Reid, 1961). An interesting episode from the epi demiological point of view started in February 1947 when a small number of outbreaks occurred in the county of Somerest. Slaughter of affected flocks was slaughter
si,
NUMBER OF OUTBREAKS OF NEWCASTLE DISEASE IN GREAT BRITAIN AND LANCASHIRE, 1946-1961
1946
'47 '48 '49 *50 '51
'52 '53 '54 '55 '56 '57 '58 '59 '60 '61
(Modified from Collender, 1958)
Figure 17.
control
of Newcastle
eradication
As
disease rather than
should be the immediate
a result, the
aim.
voluntary use of killed vac
cines was encouraged
and, although
the
disease was still reportable under the Fowl 82
Pest Order, the slaughter of flocks affected less acute forms of the disease ceased in March 1963. Slaughter was to be
with the
used only should the peracute form of the disease re-appear.
Control procedures have been adopted with very satisfactory re
in Switzerland sults
1962,
(Hess,
1963).
These
proce
dures have included :
— The testing ported
of random samples of im poultry for the presence of
Newcastle disease virus.
— The — The
cination. slaughter
of all infected
1960 (Hess,
1963). Switzerland is frequently through
large
Compulsory vaccination virus
vaccine
reduction
using
live
a
has led to a considerable
in the number of infected farms
in Yugoslavia (Fiolic,
1957). To accom eradication of the disease
plish complete in that country,
strict
control
measures
exposed to in
importations
and the fact that the national
of
flock of six
million
fowls was free from Newcastle
disease
in 1960 has been attributed largely
routine examination of samples of poultry carcasses (Hemsley,
imported 1961).
France
South Africa The incidence of Newcastle disease and the control
measures adopted in the Re
of South Africa have been sum marized by Anon. (1950b) and Kluge (1964). The first outbreak was diagnosed in the Durban area in 1944 (Figure 3). Outbreaks which were identified in 1944, public
1949, 1950, 1951, 1953, 1954, 1961 a slaughter
compensation
the slaughtered
in 1956 (Lind-
have been enforced.
slaughtered poultry (18,400 tons in 1961),
three-quarters
break in Sweden occurred
Yugoslavia
The testing of samples of imported for the presence of Newcastle disease virus was begun in 1947. This procedure was associated with a progres sive decline in the number of outbreaks in the country. Field outbreaks ceased in
the
in 1947, control measures,
have eradicated the disease. The last out
flocks
poultry
In France,
lentogenic
after diagnosis has been
established.
to the
and
have resulted
gren, 1963).
immediately
fection
virus
break occurred
prohibition of live imports. prohibition of any form of vac
— The
strains of in the abandonment of an eradication policy based on compul sory slaughter and the payment of com A very (Fritzsche, 1963). pensation similar situation occurred in The Nether lands in 1950 (Hoekstra, 1961). By way of contrast, in Sweden, where the first out
of mesogenic
Switzerland
paid
policy, did
in which
not
of the commercial
exceed value of
birds, was hard to enforce
(Fontaine, 1963). This was because many farmers failed to notify the authorities of the existence of the disease, and because diagnosis was difficult when a strain of virus of low virulence was involved.
Germany, the Netherlands and
Sweden In western Germany the rapid spread of Newcastle disease and the appearance
1962 were all of the velogenic high
mortality
and
were
and
type with
eliminated
by
slaughter and other strict control measures
(Anon., 1950b). These measures included the designation of infected areas; the con trol of movement of poultry and poultry products;
quarantine
and
disinfection.
Voluntary vaccination, including the use of live vaccines, was permitted within the infected area. Vaccination teams worked under
official
the mild
supervision.
(lentogenic)
In July
1960,
type of Newcastle
disease was diagnosed in South Africa for the first time (Kluge, 1964). This form of the disease was eliminated by 1961. 83
In Canada, Newcastle
An
disease was first
of On
in 1 948 in the province
recognized tario.
policy was immedi
eradication
ately enforced under the Animal Contagi ous Diseases Act, and during the follow ing three years only a small number of outbreaks were identified in the province. However, in 1951 and 1952 the number of outbreaks increased to a total of 68. In the province of British Columbia, New castle disease first appeared in February 1950 and spread very rapidly. The peak of the epizootic occurred in May 1950
In
and 133 flocks were involved. provinces
of Canada,
isolated outbreaks
A
sures failed to eliminate result,
federal
supported
The reasons for the termination of the slaughter policy may be sum marized as follows (Lancaster, unpub lished data) :
— The
certification
requiring live
ported
additional
poultry
and
ported live poultry and hatching eggs.
—The
difficulty of identifying the mild form of the disease in the field. — The favourable progress made in the disease of Newcastle development
and
vaccines.
— The
measures:
that
all
hatching
risk of
to the continued
the introduction of the disease in im
have occurred.
two
exposure of the Canadian poul
try industry
slaughter
by
policy was discon
the slaughter
Canadian
the other
only sporadic
mea
the disease. As a
tinued in 1954.
policy was carried out in Canada for six years: from Febru ary 1948 to March 1954. This policy was one
was dense, control
try population
Canada
especially the wide
industry,
spread
movement
hatching
im
of the Canadian
characteristics
poultry
of live
poultry,
and poultry carcasses,
eggs
which did not assist disease eradica
eggs
tion procedures
from flocks free of Newcastle disease; the second requiring the cleaning and disinfection of poultry crates after each use (Wells, 1948). In certain prov inces, eradication of Newcastle disease
was terminated
was achieved; but in areas where the poul
Contagious
originate
based on slaughter.
Although the official
slaughter
castle disease has continued in the regulations
policy
in 1954, New
in Canada
to be named
made under the Animal
Diseases Act.
ACTION OF CHEMICALS ON NEWCASTLE DISEASE VIRUS A been
number adopted
of different to
evaluate
activity of disinfectants
methods the
have
viricidal
against Newcastle
that the virus
(usually
as
crop
and by 1:10,000 of potassium permanganate. One
disease virus. Using an emulsion of mouth
per cent formaldehyde)
from infected chickens as the source of virus, Doyle (1927) examined the effect of a number of disinfectants
solution
(Table 11) and concluded that Newcastle
with 2 to 3 per cent creolin
exudate
diluted
contents) was inactivated after 30 to 40 minutes by 1 and 2 per cent formalin (37
procedure
used successfully
in the field
disease virus was more susceptible to the
daily solution and to supply the poultry with drinking water
action of alkali than to that of acid.
containing
Kohn (1958) showed that when New castle disease virus was brought tact with gizzard
contents
at
into con
pH 2.6, its
viability was considerably reduced. Stud ies reported by Farinas (1930) showed 84
was
to disinfect
the
environment
1 : 2000 potassium
permangan
(Farinas, 1930). Similar recommenda tions for field control have been made by Johnstone (1931) and Haddow (1941). Tilley and Anderson (1947) used a ate
method whereby test solutions were mixed
TABLE
— Effect of Disinfectants on Newcastle Disease Virus (Doyle, 1927)
11
Minimum strength required to kill the virus after contact for one hour at room temperature (1 5'C)
Disinfectant Methyl alcohol Ethyl alcohol Ether
Acetone HCI NaOH Antiformin Formalin Mercuric chloride Oil of cloves Carbolic
1
2
1 1
2 5
1
2
Not killed by
1
25
Not Not Not Not
1 1
N/50 killed
by
killed by killed by killed by
1 1
Potassium permanganate Copper sulphate Hydrogen peroxide
1 1
Lysol
1 1
Cresol
1
amnio-allantoic fluid
of virus. The mixtures
as
the source
were held at 20°C
and, after 5 minutes exposure, aliquots were removed and inoculated into embryonating adopted
A
similar procedure was by Cunningham (1948), except
eggs.
that a 3-minute
reaction
time was used.
When comparing his results with those of
Tilley and Anderson (1947), Cunning ham
( 1 948 )
sodium
indicated
that
1 per cent
hydroxide,
solis saponatus,
100 100 20 500 5,000
1 1
Izal
with
100 50
and
3
2
per
cent
liquor cre-
per cent
phenol
were effective against the virus during a 3
20 2
5,000 1,000
reported. When Asplin (1949) studied the action of for
similar to those previously malin on Newcastle that a virus
disease virus he found
suspension
after exposure
remained
formalin for one hour at 18.3°C (Table 12). On the other hand, Beamer and Prier (1950) have reported that 0.5 per cent formalin inactivated the virus in 30 min utes at room temperature. A different method of assessing the ef fectiveness of distinfectants has been re ported by Reuss (1957). He placed hy podermic needles contaminated with New
5 minute reaction period. Similarly, Weidenmuller (1951) concluded that 2
castle disease virus-infected
per cent sodium
in the disinfectant
to
tive
hydroxide
was an effec
On the other hand, (1948) reported that the destruc
disinfectant.
Moses
tion of Newcastle cent sodium
disease virus by 2 per
hydroxide
active
to 2 per cent commercial
allantoic
fluid
and then inserted them
into embryonating eggs. Using this meth od, Reuss (1957) found that the virus was inactivated
by
a 10-minute
exposure
to 3 per cent formalin solution.
was irregular.
Asplin (1949) generally used a mini virus-disinfection reaction time of one hour. This precluded a direct com parison being made with other results Nevertheless, (Anon., Asplin 1959).
Another method of evaluating disinfec tants was studied by Haussman and Grafe
(1949) concluded
virus haemagglutinin.
mum
that his results
were
(1957) who reported close relationship and the action
that there
was
a
between viricidal action on the Newcastle
disease
Accordingly, it
has 85
TABLE
12 —
Action of Formalin on Newcastle Disease Virus (Asplin, 1949) Time of exposure
Concentration of formalin
Result
Temperature
'F
per cent 10 days 16 days 50 days 90 days
34-35 34-35 34-35 34-35
Inactive
1 hour 12 hours 12 hours
65
Active
65
Inactive
65 98 98
Inactive Inactive
0.1
1 hour 1 hour
0.1
6 hours
98
0.2 0.1 0.05
0.025 2.0 2.0 1.0
0.25
been suggested that disinfectants evaluated
on
might be
the basis of reduction
haemagglutination
in
Fumigation with Formalin Using five different test materials con with Newcastle disease virus, Beamer et al. (1949) showed that, in a with fumigation forced draft incubator, 35 ml. formalin released by 17 gm. potas sium permanganate per 100 cubic feet of taminated
space was sufficient to inactivate the virus during
a period of 2 to 3 hours. In thick fluid, destruction
films of yolk or allantoic
of the virus was achieved with 70 ml. formalin and 34 gm. potassium perman ganate per 100 cubic feet of space. Beamer et al. (1949) have emphasized
that, to pre
from protecting
vent thick deposits
virus, all contaminated
the
surfaces in the in
Inactive Inactive
Active Inactive
gation of brooder
fumigation
Mansfield
in
various
locations
castle
disease virus
adsorbed
onto
sur
malin
and potassium
been recommended
permanganate
by Nobrega
has
(1955). The value of formalin alone in the fumi
86
in
New placed
rooms
the
and
Other Disinfectants The distinfectant properties of a solution of the amino acid di(octyl-amino-ethyl)glycine has been examined.
found
a 2 per cent
infectant
inactivated
5 per cent solution
faces. The use of smaller amounts of for
on egg shells
incubators.
Greuel (1963a) has con cluded that exposure for 5 to 10 minutes to 30 ml. formalin released by 20 gm. activate a considerable proportion of New
by
(1950).
inactivated
apparently
castle disease virus
virus
potassium permanganate is sufficient to in
rooms as
has been reported
They treated approximately 5,500 cubic feet of space with 8 U.S. gallons of 37 per cent formaldehyde solution. When the tempera ture of the building was raised to 37.8"C and the humidity to 100 per cent, the This time was 20 hours. fumigation and
cubator should be cleaned thoroughly be fore fumigation.
and incubator
well as incubators Schmittle
titre.
Inactive
Reuss (1962) of this dis
solution
Newcastle
disease
in test tubes in 20 minutes;
a contaminated
and a
inactivated the virus on
hypodermic
syringe with
in 15 minutes (Reuss,
In
tests
where
closed composition Newcastle
1963). disinfectants
of undis
have been used against
disease virus in different media,
the viricidal action has been reduced in the presence of albumen (Grafe and Haussmann, Grafe, and 1956; Haussmann 1956). There is much less protein in virus.
suspensions in allantoic fluid than in organ Haussmann, and suspensions (Grafe 1957).
Certain chemicals
have been found to
have little or no viricidal action
against
Thus, neither ascorbic acid nor cysteine hydrochloride at a concentration of 0.2 mg. per ml. has Newcastle
disease
virus.
had any effect on the virus Datta, 1950a). In contrast, centration serum
(Sinha and a
low
con
lecithin-like fraction from of inactivating disease virus at 37°C (Utz,
of
a
has been capable
Newcastle
Using both in vitro and in ovo methods, McLimans et al. (1957) tested a variety of compounds containing a terminal o o
-
hydroxy-aldehyde
against Newcastle
grouping
disease virus.
One of the compounds,
designated "Keth-
oxal," was found to be
a potent inactiva
ting agent in vitro against the virus. Lyohas been inactivated
after
4 hours exposure to ethylene oxide
virus
(car-
philized
boxide) under a pressure of 1,500 mm.; while wet virus has been inactivated in 3 hours
1949).
or
ketoaldehyde
(Mathews and Hofstad, 1953).
STERILIZATION OF ATMOSPHERES CONTAMINATED WITH NEWCASTLE DISEASE VIRUS Robin (1962) has reported that, under experimental taining
conditions,
volatile
an aerosol acids
organic
periods of 3 minutes
used
con
for
each day was suffi
cient to prevent the air-borne transmission
of virulent Newcastle
disease
virus
to
susceptible chickens. disease virus of of a glycol mixture containing
The effect on Newcastle an aerosol
a quaternary
ammonium
compound
was
shown
that the disinfectant
aerosol
was
not harmful to hatching eggs or chicks.
Similarly, in a broiler flock, the use of of triethylene glycol has ap peared to reduce the spread of Newcastle disease (Ellis et al., 1952). However, adequate concentration of the vapour may be difficult to obtain under general farm conditions. The ultraviolet irradiation of the air of a poultry house by means of an aerosol
examined by Walker et al. (1953). First a fog of the disinfectant was produced in a chamber; then a fine virus suspension
of infection
was
blown in. Air samplings taken at inter from 10 to 120 minutes showed no evidence of live virus. Furthermore, it was
teries,
vals
hens housed on other floors in the same
did not prevent
"Sterilamps" Newcastle
during
a natural
the spread
of in bat
outbreak
disease among broilers
and the disease spread
to laying
building (Levine and Hofstad, 1947).
CONTROL WITH HYPERIMMUNE SERUM Early studies on the prophylactic therapeutic value of hyperimmune
and serum
have been reviewed
of appreciable quantities of (Farinas, 1930; Haddow, 1941). To meet this problem, Mitchell and Walker (1951b) and Spalatin (1948) horses and they found hyperimmunized production immune
serum
by Beaudette (1943). majority of these studies involved small numbers of fowls under field or laboratory conditions. In general, results
the antiserum
were not encouraging.
ing
The
One
of the difficulties
has been
the
power
had considerable
when
virus in chickens.
neutraliz
tested against
virulent
Less satisfactory
results 87
have been reported by Cooper ( 193 1 ) and (1955). Moynihan et al. (1954)
Lulic
that 0.5 ml. of antiserum
found
adminis
tered between 24 hours and 72 hours after virus exposure failed to prevent the devel opment of Newcastle disease.
from goats (Anon., 1943 ) ; it has delayed death of inoculated chickens for only two to four days Immune
has not
prepared
very
satisfactory
et al.,
(Placidi serum
serum
been
1952). However, a potent been prepared from goats
has
(Fomina and Ochkina, 1951) and also from a calf (Coronel, 1939). Hyperimmune serum has been prepared in fowls and turkeys by injections with virulent virus (Majewska and Zebrowski, 1955; Nai and Garinei, 1945; Skoda and Zuffa, et al.,
al, 1956) and antibiotics (Nobili
1956a; Zuffa et
has been treated with
1960). Immune serum in amounts 1 ml. has protected fowls against
of 0.5 or
virulent virus administered
simultaneously
( Zuffa et al., 1956) , or 48 hours previous
ly (Nai and Garinei, 1945). Although the cost of this type of serum has sometimes been considered prohibitive (Anon.. 1943), satisfactory results have followed its use in infected flocks; and mortality has been considerably reduced (Capaul et al., 1963; Lulic, 1955), or has ceased 6 to 10 days after treatment (Nobili et al., 1960).
(1948) and Tanasugarn
Spalatin mune
serum
anticipated
has
or
1963) globulin
serum
simultaneously
1960). It
has
can transfer
munity
to
Wogan,
1963).
been
their
and
gamma
(Vasington
et al.,
that hens
to Newcastle
disease
of this im
a portion
offspring
to
Luginbuhl,
also been shown
passively immunized virus
have and
(Wills
immune
the
flocks.
in infected
procedures
egg yolk
reduced
markedly
mortality
Alternative inject
(1961) of im
that the injection
have reported
(Grun
and
CONTROL WITH HYPERIMMUNE SERUM COMBINED WITH VIRUS This method of immunization has been (Coronel, 1939; unsatisfactory
generally
1943;
Iyer,
Zuffa
though
Seetharaman, and
Skoda
1951b),
al-
that the intranasal serum
of immune
instillation
with
simultaneously
virus resulted in satisfactory
attenuated immunity,
(1959) found
CONTROL WITH ANTIBIOTICS AND OTHER MEDICINAL AGENTS The therapeutic effect of antibiotics in of Newcastle disease infection has been reported. Giovaneli (1962), for
the control example,
found
that
symptoms
disap
peared and mortality ceased after injection a mixture of three antibiotics. Penicillin
of
administered proved 1952).
of
intramuscularly some
However,
value
has
also
(Michalewicz,
the oral administration
of oxytetracycline has had no influence on the course of experimental Newcastle disease, although it has reduced mortality in chicks — probably because of the in 88
hibition of secondary 1960; Tanasugarn,
(Reuss,
invaders
1961).
Substances which have shown no curative
little or
value against the disease in
clude: xenalamine
(Gagliardi and Girotto, (Anon., 1943),
1961),
sulphapyridine
vitamin
A
(Bonaduce,
1950a)
and corti
(Hababou Sala, 1960). In con trast, increased riboflavin supplementation costeroid
has reduced mortality
10 and 17 per cent
in groups of artificially infected (Squibb,
1963).
chickens
CONTROL BY VACCINATION In
of national control mea
a summary
sures in effect during
out of
that
1961, it was shown
countries
103
reporting
85 had adopted vaccination
disease,
the
as the
control procedure (Table 2). The of vaccination are usually borne by the poultry industry. Vaccination of the United States broiler crop in 1956 cost more than four million dollars (Hanson and Brandly, 1958). In Africa, the cost main
costs
vaccination
of
has generally
been
con
out of proportion to the economic
sidered
of
value
the
(Kaschula,
stock
1950;
1954; Rao and Agarwal, 1962; Thompson and Osteen, 1952; Vandem
ada,
aele, 1961). There are some exceptions: in Spain (Blanco, 1949), the United States (Flowers et al, 1960) and Guate mala (Correa, 1963; Correa and Rosales, 1961) vaccines have failed to protect against an indigenous field virus, especi ally when the field virus has been highly ever, it is important tween
vaccine
breaks
(Jungherr and Markham, 1962).
breaks
1961) and the demand for vaccine has been poor ( Winmill and Haig,
The latter involve
1961).
vaccine
Vandemaele,
Compulsory
vaccination
has sometimes
(Fiolic, 1957) in spite of (Fontaine, 1963; Winmill and Haig, 1961). To overcome the difficulties, the cost of the vaccine has been subsidized (Garside, 1962). Studies of vaccination for control of the disease under methods of varying adopted
been
husbandry were begun soon after identifi cation of Newcastle the literature
disease in 1926
and
on the subject is now con
1964a). To review is beyond the every report on vaccination (Lancaster,
siderable
of this publication; nevertheless, an is made in the following pages to various facets of the subject.
attempt survey
Gualandi
Gill
et
However,
(1949), Traub (1944), Upton
several strains of vaccine virus in widely dif
have been used successfully
geographical
clinical
different
regions
and
against
manifestations of the 1959; Jungherr and
(FAO-OIE,
Markham, and
handling
(Davis
factors
many
relating
and
administration
of
al,
1951; Larose
and
et
1959; Marek; 1957; Tenni-
Van Roekel, son, 1963).
1962; Marek,
1957; Mitchell
Walker, 1953; Pagnini,
Procedures for Evaluating Immunity When vaccination
was first adopted, the
potency
of vaccines
could
assessed
because
disease
the
be
readily
frequently
caused over 90 per cent mortality. How ever, in certain countries, Newcastle dis ease has since changed to a
form in which
mortality is considerably lower (Fritzsche, 1963; Jansen and Kunst 1952; Lissot, 1956; Reid, 1961 ; Skoda and Zuffa, 1958), or the nervous symptoms more numerous
(Jaksic and Stefanovic, 1957). In these can, reduction of mortality
therefore,
al (1959),
etal. (1953) and Valdes Ornelas (1964) have led to a questioning of the feasibility of using any one vaccine in all situations.
disease
the
vaccination
countries,
Antigenic Plurality Results reported by
ferent
to
and
difficulties
serious
scope
1964). How to differentiate be
(Valdes Ornelas,
virulent
1950; Ques-
no longer
be
used as the sole
for the evaluation of vaccines. Moreover, resistance to infection of the
criterion
respiratory temic
epithelium,
infection
as
to sys
immunity
denoted
clinical
by
signs, and resistance to a decrease in egg production
may be independent
other
(Bankowski 1962b; Markham
and et
of each
Corstvet,
al,
1951a,
1960, 1957;
Raggi and Lee, 1962),
or show a lack of
correlation
et
(Bankowski
an alternative
procedure
munity, an intradermal
al, 1958b). As to determine im
test has been used 84
(Wasserman
and Yates, 1953; Yates et
Probably
a
still
better procedure
antibody
for
develop or decline at the same rate follow ing vaccination (Keeble and Wade, 1963;
is to expose vacci
determining
immunity
nated
non-vaccinated
and
chickens
to
chickens which have been in fected with a virulent field virus (Bankowski and Corstvet, 1962a; Mazzaracchio susceptible
and Orfei, 1956; Taylor, 1953). However, in this procedure the environmental tem perature (Francis and Kish, 1955; Sinha
al,
et
1957),
as
well
as
other
stress
factors
(Schultz and Feiling, 1954), can influence the resulting mortality. Vaccin ated birds, when exposed by injection or by contact to virulent virus, may show no mortality or sign of infection but may experience
al,
a temporary
viraemia
(Gill
et
1959; Hofstad, 1956) or a respiratory infection (Doll et al, 1950c, 1951b; Ban-
kowski et al, 1957). The use of the haemagglutinationinhibition (HI) test as a quantitative measure of immune response has already
been reviewed on pages 61 to 63. It must be emphasized that the HI response can not be compared directly with the immune status as measured by challenge with viru lent virus (Doll et al, 1950a, b, 1951a; Hamann, 1958; Hitchner and Reising,
1953a; Hitchner et al, 1951a; Ileri, 1950, Levine and Fabricant, 1952; Mark ham et al, 1954; Mazzaracchio and Orfei, 1955; Raggi and Lee, 1962; Simmins and
Baldwin, 1963; Valadao, 1955; Winterfield and Seadale, 1957; Winterfield et al, 1957). Schmidt and Schmidt (1955) have reported that 8.5 to 95 per cent of fowls with vaccination
HI
and
inhibition, virus neutralization and speci refractivity to infection as a single entity (Brandly et al, 1947; Dardiri and Yates, 1962; Hanson et al, 1950; Schmittle, 1953). Thus, Karzon and Bang ( 1951 ) showed that the neutralization test fic
in
The fact that serum neutralizing 90
(SN)
and
embryo
the
haemagglutination
inhibition test yielded parallel results in the measurement of antibody during early convalescence. Later in convalescence, the
HI
titres were lower.
No definite relationship has been found between HI and SN titres (Nakamura et al, 1956) and respiratory infection (Levine and Fabricant, 1950). However, the SN test has been thought to give a truer picture of immunity than the HI 1951; Hitch test (Atanasiu and Gareau, ner and Reising,
(Gill
weeks
1954),
especially
when
period extends beyond 50
the observation
and
could, therefore,
Stone,
1964),
and
it
be useful to supplement
HI test (Crowther, 1963). It follows that the methods and criteria for evaluating the immunity adopted
the
engendered by Newcastle have
been
Johnson vented
et
a
( 1 960)
disease vaccines
(Hofstad,
variable
1953b;
al,
1954) and this has pre close comparison of all the
results reported.
Bankowski and Corstvet
were of the opinion that immunity
in Newcastle
posure
for different vaccine strains.
Schmidt, 1959) to regard the immune
bodies associated with haemagglutination
Crowther (1963) has suggested that, be cause different strains of Newcastle dis ease virus vary in their ability to stimulate the formation of HI antibody, the HI test not be used as the sole basis of
1960;
makes it impossible
measurable
should
do not always
antibody
Lee,
and
Raggi
titres up to 1:16 died after experimental infection; whereas fowls with vaccination HI titres of 1:32 and over resisted infection. Furthermore,
comparison
HI
al,
1954).
disease consisted as
well
of many
unmeasurable
as
factors.
In the discussion emphasis
is placed
follows, more
that
on the results of ex
of vaccinated
birds
to
virulent
virus and results obtained in the field than on serological responses. criteria,
it is recognized
In
adopting these
that a drop in
egg production or the presence of respira
tory symptoms would be a more critical test of immunity than mere survival
in
No.
1
1
1
following
Period
showing
et
Disease
1
365
1
:16
of chickens
(Lancaster
Newcastle
1
33
tested
3
2
1
1 2 3 4
(Hofstad,
In this
1953b).
connection,
Bankowski
(1961a) found that the level of immunity which protected hens against a drop
in egg production
was related to
the amount of virus in the vaccine. How ever, during declining immunity, transi tory respiratory symptoms and a slight drop in egg production have appeared to be unavoidable
when vaccinated birds are
exposed to virulent field virus (Garside, 1962; Hitchner and White, 1956). Never theless, significantly fewer soft-shelled
following the
have been produced
eggs
challenge of vaccinated
of unvaccinated
ing the challenge
(Gill
birds than follow birds
hand,
administration
of virus
of
a virulent
in
(Lancaster et al., 1960). In young chicks, the degree of immunity (Doll et al., 1950b; Richey and Schmittle,
or of virus multiplication in
1962),
the
oral mucosa (Gagliardi and Irsara, 1958), has been directly influenced by the amount
of
and Stone, 1964).
the
drinking water has resulted in satisfactory immunity in chicks possessing maternal antibody (Gagliardi and Girotto, 1960). The results given in Table 13 indicate that satisfactory HI serum titres do not result from the intra nasal vaccination with Bl virus of chicks at 2 days of age parentally-immune strain
yolk-transmitted
the
and
antibody,
of the vaccine virus (Gagliardi and Girotto, 1960). As a result of the use of the B 1 vaccine during a five to six year period, with resulting passive immunity in young chicks, the overall duration of im munity has apparently been reduced from virulence
Factors that Influence Development of Immunity Passive lmmunity The literature
on the transmission
maternal antibodies
of
to the chick has been
reviewed by Beaudette and Bivins (1953) and Brandly et al (1946b) and it has been discussed
by Hanson (1957) and Grun and Wogan (1963). At hatching time, chicks from immune dams have shown a much lower serum that of the dam. Bornstein
HI
titre than
et al.
(1952)
found that this titre in the chick rose to a peak at 3 days of age and progressively declined thereafter.
Usually, congenital passive immunity or the administration of immune serum has interfered with the development of active immunity disease
in response to Newcastle
vaccine
(Bankowski 1952; Doll
given
intramuscularly
et al., 1958a; Bornstein
et al.,
et al., 1950b; Keeble and 1963; Keeble et al., 1963; Markham et al., 1954), subcutaneously (Haig
Wade, et
al.,
1962;
Zuffa
and
Skoda,
1959),
(Bankowski and Corstvet, 1962a; Richey and Schmittle, 1962) or in drinking water (Marek and Raszewska, 1959; Winterfield and Seadale, 1956b; intranasally
Winterfield 92
et
al,
1957).
On
the other
12 weeks
to 6 weeks
et al.,
(Bankowski
1957).
Usually, the
response
best
in
chicks
immunity of a live has followed administration vaccine by the respiratory or conjunctival routes (Beaudette and Bivins, 1953; Born stein et al, 1952; Hitchner, 1950; Hitch ner and Reising, 1953b; Markham et al., 1951a, b, 1954; White et al, 1953; Zuffa and Skoda, 1959). In passively immune day-old chicks, intranasal instillation has having
maternally
transferred
resulted in a better immune response than that obtained by aerosol vaccination
(
al,
Rao
1963). Such findings have led to the be conclusion that to delay vaccination et
cause of maternal immunity is not justi fied (Crawley, 1954; Hitchner et al, 1950;
Markham 1953). vaccine
al,
et
1951b; when
However, has
White et al,
an
inactivated
been used, better immunity
has resulted when chicks from immunized hens
were
first
vaccinated
at
8
days
(Keeble et al, 1963) or, preferably, 21 days of age (Keeble and Wade, 1963). The value of passive immunity in pro tecting
young
birds
has been examined.
A
number
of reports have indicated
absence of protective
the
in young
immunity
chicks from vaccinated flocks (Doll et al, 1951a; Monti, 1954; Olson et al., 1950; Reuss and Hilbrich, 1960; Zureck,
1958). Some of these results can be explained by a
low level of immunity
(Doll
flock that,
in the parent
al,
1950b) or by the fact congenital antibodies give
et
in chicks,
of susceptible in a durable
chicks
day-old immunity
can result 5 months
lasting
(Asplin, 1952; Lancaster, 1957b) to 12 months (Hitchner, 1950). It is believed that at about 1 to 2 months of age, chickens are sufficiently an optimal
response
kowski and Rosenwald, Rao,
1959b;
developed to give
to a vaccine
Raggi
(Ban
1956; Gupta and Lee,
and
1962;
and Fabricant, 1 950) . In contrast, a num
1959) or other antigen (Wolfe and Dilks, 1948). When considering the influence of age,
ber of authors
it should
little protection against
from
to the respiratory
Newcastle
immune
disease virus
have
found
tract
(Levine
that chicks
hens resist artificial
infec
Schmidt,
and Walker, 1953; Russeff
disease
Miteff, 1957) or develop asympto matic infection (Doll et al, 1951a), de
mature
tion (Mitchell and
of the challenge
pending on the virulence
be mentioned
evidence
that
that there is some to
resistance
Newcastle
as birds progressively (Baldelli, 1957; Brandly et al,
increases
1946c; Cole and Hutt, 1961;
Gill
et
al,
1959).
virus.
This resistance due to passive immunity for varying periods up to about 4 weeks (Anon., 1962a; after hatching
Virus Titre of the Vaccine
lasts
al, 1946b; Christie et al, 1963; al, 1949; Garinei, 1945; Hitchner et al, 1950; Maglione and Dotta, 1957; White et al, 1953). However, field Brandly et
Clancy et
studies
have
shown
that
in
heavily
fected areas passive immunity
in
cannot be
for the protection of 4 weeks of age (Hitchner etal, 1950; Levine and Fabricant, 1950). depended
upon
chicks under
It must be emphasized that in many of the reports it
has
not
referred to on pages 95 to 115 been
possible
whether the chicks mune
or
particularly
to
determine
used were from im
susceptible
parents.
This
is
true of reports of field results.
In addition to the antigenic characteris tics of the vaccine, the titre and the dose
of virus play an important part in the level of immunity produced (Bankowski and Hill, 1954; Brandly et al,- 1946a; 1954; Hitchner and Reising, Crawley, 1954; Raggi and Lee, 1962, Winterfield and Seadale, 1 957). Thus, under field con ditions, White-Stevens
1963). Bankowski and Corstvet (1962b) found that a single injection of a vaccine
do
not
show
the
maximum
(Bankowski et al, al, 1946a; Doll et al,
response to antigens
Brandly et 1950b; Hitchner et al, 1950; Keeble and Wade, 1963; Mansjoer, 1961; Nakamura et al, 1956; Waller and Gardiner, 1952; Wasserman and Yates, 1953; Wolfe and Dilks, 1948). Nevertheless, vaccination 1957;
ELD
at 10~7
per 0.2 ml. gave nearly the
level of immunity
same
as two
doses of
ELD administered nine For a water-administered
vaccine of 10^4,5 apart.
vaccine,
Chicks, even in the absence of passive immunity,
was directly
related to the dosage of virus particles or the amount of antigen (Keeble and Wade,
weeks
Age at Time of Vaccination
(1961) considered
that the degree of immunity
the
ELD
minimum
titre
should
be
(Winterfield and Seaof dale, 1957). For spray administration 10-7
Bl
per ml.
virus, the titre should be between 10^7
and 10-8
ELD
per 50 ml. (Crawley, 1954).
Viral Interference The different features which comprise of viral inter phenomenon ference and viral infection of the cell have the overall
93
been
discussed
in detail
by Goret and (1964). Hanson and Alberts (1959) and Han son et al. (1956) showed that when a virulent field strain of Newcastle disease virus was administered simultaneously with
in suspended
Provost
infectious bronchitis virus by the intranasal route, there was interference with the sub
of Newcastle disease Other workers have found that the interference resulted in poor immunity to the infectious bronchitis vaccine sequent development infection.
(Bankowski et al, 1955). Raggi et al. (1963) obtained similar results in chick embryos. An homolagous interference has been established
in chicken
cells
by the use
of ultraviolet-irradiated Newcastle disease virus. As a result, the cells lost their ability to adsorb active virus. It has been suggest ed that this type of interference occurred at the cell surface (Baluda, 1957, 1959). An
strain of Newcastle disease virus has also inhibited the growth of a strain of the same virus adapted to the brains of new-born mice (Sinkovics, egg-adapted
Morimoto propagation
et al.
(1962) found that the
of Newcastle disease virus in
certain
cell
Russian
spring-summer
cultures
was
inhibited
encephalitis
by virus
and by Japanese encephalitis virus. In terference between other heterologous viruses have included: the interference in birds inoculated with a mixture of attenu ated fowl plague and Newcastle disease
(Daubney and Ishak,
1953); the between western equine en cephalomyelitis and Newcastle disease interference
viruses when propagated fibroblast monolayers
in chick embryo
(Levine, 1958, the interference between mumps virus and Newcastle disease virus in embryonated hens' eggs (Sinkovics, 1957a); 1962b);
the
interference
Newcastle cultures
between
influenza
and
disease viruses in chick-embryo
(Tyrell,
1955); and the inter
ference by ultraviolet-irradiated
Newcastle
disease virus and vesicular stomatitis virus 94
ease
the
L
cells (Cantell
a mesogenic
vaccine
et al., 1962).
live Newcastle
administered
is
dis
to birds
in
of infection, recovery frequently follows, or further spread of the disease is stopped (Harnach and Polak, 1964; Mihalka, 1963). This interference or cell block has been associated with the use of the Mukteswar vaccine (Daubney and Mansi, 1948; Generoso and Menearly
doza,
stages
1950)
control
of
and
been used in the
has
of outbreaks
a number
(Karc-
zewski et al., 1955). Interference between the Mukteswar vaccine virus and field virus has been demonstrated to occur between 20 and 72 hours after vaccination
(Gupta and Rao, 1959a; Haddow and Idnani, 1946; Karczewski et al., 1955; Nilakantan et al, 1960a; Russeff, 1956). A similar Newcastle disease viral inter ference has been observed tures (Durand, 1961).
in tissue cul
Protection against virulent virus has been demonstrated one to three days after vaccination with the Hertfordshire virus and
(Buzna
1957b).
viruses
When
1952; Schmidt,
Hodosy,
1951;
Gualandi,
1952); after six hours with
another mesogenic vaccine strain (Jezierski, 1953) and after two to four days with a
formolized vaccine
(Brandly
al,
et.
1946a).
With lentogenic strains of Newcastle viral interference has not always been demonstrated. Thus Doll et al. (1950a), Hitchner and Johnson (1948) and Nilakantan et al. (1960a) found disease virus,
vaccine
strains
protection virulent
when virus
Bl
and
birds soon
F
afforded
were after
little
exposed
to
vaccination.
Similar results have been observed under field
conditions
(Lancaster,
unpublished
data). In contrast, it has been suggested that the vaccination of young chicks with Strain F virus intranasally (Rao and Agarwal, 1960), or Bl virus intraocularly (White and Appleton, 1953), results in protection due to viral interference 48 hours later.
Differences Between lndividual Birds In any group of vaccinated chickens, every individual develops a satis factory immune response (Asplin, 1952; not
1962;
Garside,
et al.,
Lancaster
1960),
regardless of the kind and quality
of
the
immune
varies
response
between individuals; and approximately
per cent of susceptible
to 7
shown
an
abnormally
birds
poor
5
have
response
(Markham, 1962). Differences immune
between individuals
response
to
in their
Newcastle
antigen may be influenced
disease
by a deficiency
globulin (Hanson, 1957); by of the chicken background (Millen, 1960); or simply by the varying
of gamma the
genetic
ability
of
different birds to respond to the
(Ram, 1961).
virus antigen
Effect
of Vaccination on Susceptibility
to Another
Disease
Hanson and Alberts (1959) and Han son et al. (1956) have shown that chickens
exposed to small quantities disease
of Newcastle
virus may be more susceptible to on re-exposure. This viral
the same virus
sensitization vaccination
may failures
play
a role
leucosis
in
some
(Hanson, 1957).
Another feature relevent to Newcastle
and
moderate
leucocytosis
between
the
(Consoli
et al.,
1955).
A
in the degree and duration
reduction
of immunity has been associated intercurrent
infectious
diseases
and
with in
(Blanco, 1949; Brandly, 1948; Hoekstra, 1961); with poor condi tion (Garside, 1962; Marek et al., 1961; with and Brandly, 1953); Pomeroy and with (Schiavo, 1960); moulting (Gualandi, 1953). The last caponization ternal
parasites
mentioned
effect was not, however,
ob
served by Brandly et al. (1946a). The stress placed on poultry during transport to and from a market may result in a breakdown of vaccination immunity (Schultz and Feiling, 1954). Furthermore, the prolonged feeding of tobacco powder as a parasiticide appears to inhibit the production of Newcastle disease HI anti bodies following vaccination (Papparella, 1955). According to Squibb (1963), the addition of B complex vitamins above normal requirements does not appear to influence antibody response, as determined
HI
by
titres. Similarly, Cho (1963) has no evidence that the bursa of
reported
Fabricius plays a significant role in the of antibodies to Newcastle production
(Bl strain). In addition to these factors, a four-fold
disease virus
disease vaccination
is the fact that approxi mately 25 per cent of a group of vacci nated fowls have shown an erythroblastosis
complex
Other Factors that Affect Immunity
of the
antigen (Bankowski and Corstvet, 1962b; Markham et al., 1951b). Furthermore, the speed
fourth and eighth day after vaccination. Thus, vaccination may act as a predis posing factor to diseases of the avian
increase in Newcastle has
been associated
disease
HI
antibody
with maximum
production (Markham
egg
et al., 1956a).
TYPES OF VACCINES— ADMINISTRATION AND EFFECTIVENESS Live and Inactivated
kinds: live and inactivated. The inactivated
Vaccines Compared
vaccines were the first to be studied and the results obtained by
Newcastle
disease vaccines
are of two
Beaudette
1 95 1b ) ,
have been reviewed 1948b, 1949a,
(1943,
Brandly et al.
( 1946b ) ,
Fabricant 95
(1951). However,
(1956) and Thompson in the early
1930's investigations
began
on the preparation and use of live vaccines (Iyer and Dobson, 1940; Topacio, 1934) and this literature Beaudette et
has been reviewed
by
al (1950), Brandly (1959),
double vaccinations
is carried out (Fabri cant, 1956). It has been suggested that a killed vaccine of high antigenicity would
give better immunity than live
(1962), Kruger Gerriets (1961) and Reis and Nobrega (1956). It has been found that under some
etal,
conditions,
was used in Spain
Fritzsche
and
and
in
areas, live vaccines
some
geographical
have certain
advan
over inactivated vaccines and this has resulted in their extensive use (Dalling, 1958; Davis et al, 1951). tages
vaccines
(Levine and Fabricant, 1952). If developed, it would probably form an essential part of any eradication
scheme
(Osteen
1961). In 1948, a formolized adsorbed vaccine
(Botija and Loizelier,
1948). Although extensive vaccination of large flocks was possible, the difficulty of small isolated farm flocks was
vaccinating
obstacle to eradication.
a major
Some 14
of live vaccines has apparently led to the elimination of Newcastle disease for significant periods of time from Cyprus (Crowther, 1952) and parts of Canada (Anon., 1962b). In
in 1962, Newcastle disease was reported to be widespread (Figure 5). Botija and Loizelier (1948) did not expect the vaccine would eradicate the disease
other
in
The concentrated
countries
it has been thought
some emergencies
of
live
vaccines
Osteen
al,
et
use
fully justified (Beach, 1961;
that
the use
1946,
1952;
Schoening
and
Thompson, 1955; Stover, personal com it is well munication). Nevertheless, that live vaccines seldom lead
recognized
of Newcastle dis (Levine, 1962a; Osteen et al., 1961;
to complete eradication ease
and Thompson,
Schoening
1955). held that
It has been generally activated
vaccines
or less durable
entirely but they believed that vaccination
conjunction with sanitary
measures
should keep it under control.
In areas of the United States, an in activated
vaccine
extensively (Beach,
has not been had
as
In
1946).
been
in
than live virus
one field study in the
United States, on premises where New castle disease was known to exist, Schoen ing et al. (1949) have reported that the of formalin-inactivated vaccine did not
entirely
enabled
the
prevent
vaccinated
birds
disease,
Pomeroy and Brandly,
by Beach
1953; Thompson,
with an inactivated vaccine.
However, available induce et
al,
1955), local mann,
96
vaccines are now certain limits,
within
satisfactory immunity (Appleton 1963; Levine, 1962a; Woernle, especially 1954)
from a (Woernle and Sieg-
when prepared
strain of virus
suitable 1955)
inactivated which,
or one found
(Hanson and when
antigenically 1951b; Koch,
et
al,
a
proper
regimen
of
a
severe infection with relatively small losses compared with the unvaccinated
has reported 1951). Beaudette (1951a) that only 60 to 85 per cent of young to mature chickens develop immunity when
but
to withstand
(Bankowski and Rosenwald, 1956; Brandly et al, 1946b; Fabricant, 1953; Hofstad, 1953b; Osteen et al, 1961;
vaccines
vaccinated
used as
anticipated
use
give a more transient
immunity
later,
years
The
advantages
associated
controls.
with
in
activated vaccines have been summarized and
(1946) and Garside of live
the disadvantages
(1962), vaccines
have been outlined by Hemsley (1962), Hoekstra (1961) and Osteen et al. (1961). Comparative tests involving either lentogenic
or
inactivated favour
mesogenic
live
vaccines
and
vaccines have given results in
of the live vaccines (Kaschula, and Fabricant, 1952; van
1950; Levine Waveren
and
1953;
Zuijdam,
Zuydam,
1953). Other studies have shown little difference between live and inactivated vaccines 1957);
(Miyamoto however
and
Schmidt
Nagashima,
(1959) found
adsorbed
vaccine to be greater than that
produced
by the
are con lento-
rather
is
it
lentogenic
cause
a
are also
between 1959; (Anon.,
ences
to
which the mesogenic
in
strains generally There
division
strains can be administered
chicks
young
an arbitrary
based principally on the fact that
a
is,
genic strains and mesogenic strains. This perhaps,
severe reaction.
of other differ
number these
two
Hanson
the need to use special
nebulizers
reconstituted
vaccine.
freeze-dried
with
viruses in The viability of Bl and drinking water adversely affected by and has been en variety of conditions hanced by the use of organic stabilizing materials (Marek, 1960; Winterfield and Seadale, 1956a). The mechanism of the immune response following vaccination by the drinking water method has been dis cussed by Baldelli (1956), Burnstein and Bang (1958), Gagliardi and Irsara (1958), and by Winterfield et al. (1957). In intranasal or inhalation vaccination with Bl virus, the virus may or may not
F
live vaccines
sidered under two main headings:
but
in
compared
main and
types
Brandly,
1955). The lentogenic strains take longer kill chick embryos and also appear unable to multiply the central nervous
a
this section
resulted
when
response
with the aerosol method. They emphasized
strain.
Live Vaccines In
durable
is
Bl
of the vaccine
instillation more
a
immune response to an inactivated
the
to the turbinate
region where
system of the chicken;
proliferates (Burnstein and Bang, 1958). However, the initial multiplication of an
if
whereas the meso genic strains readily multiply in the brain of chickens of any age introduced by route. The intracerebral of virus into day-old chickens therefore, sensitive method of dis tinguishing between these two types of Newcastle disease virus (Hanson, 1956). the intracerebral
a
principles viral
mechanical
involved
and
of for air-borne
of vaccination have been dis cussed by Bankowski and Hill (1954), (1954), Hitchner and Reising Crawley (1953a), and Markham et al. (1955a). methods
virus labelled with
a
as
and administered
spray have shown that virus
in
Bl
phosphorus
Studies using the radio-active
small
It
amounts can be detected in the abdominal air sacs 30 minutes after vaccination. has
been
concluded
that
few
virus
particles
need
respiratory
epithelium
comparatively to
reach
the
in order to stimu
) ,
( 1
)
954b
F
1
a
measurable response (Johnson et al., . Rao et al. 963 in studies with virus, showed that intranasal Strain late
on the
(Gagliardi, 1959). To relate the optimum immune ponse with methods of administering vaccines,
comparative
res
live
studies have been
sub intravenous, wing-web, cutaneous, drinking water, spray and dust. The most effective was considered to be muscular,
factors
in the preparation suitable
suspensions
strain which occurred
mucosa of the portal of entry has spread to the mucosae of other parts of the head
made of the following routes and methods: intraocular, intranasal, intratracheal, intra
Lentogenic Strains The various
attenuated
the intraocular by Kaschula (1952) and White and Appleton (1953); the intra
(1960), Hitchner and Johnson (1948), Johnson (1956), Marek et al. (1961), Markham et al. (1954), Rao and Agarwal (1960); nasal by Glinski and Szemberowa
the intramuscular by Bran et al. (1959, 1961); the intravenous by Gualandi (1949); drinking water by Johnson (1956); Wynohradnyk et al. (1958); spray by Johnson (1956) and Marek et al. (1961); and dust by Dardiri et al. (1957) and Johnson (1956). The protection afforded by mixing food
a
is,
inoculation
it
be confined
in
to
live vaccine
was considered
virus
with
the
slight by Gagliardi
(1953). 97
B1 or Blacksburg Strain
The main characteristics of this strain for its use were first reported by Hitchner and Johnson (1948) of Blacksburg, Virginia, in the United States. The initial studies showed that all ages from day-old chicks to birds in full production could be vaccinated. Within a and the indications
short
relatively
time
this vaccine
was being used extensively States (Schoening and
also
in
strain
in the United
and Thompson, 1955) parts of the world
other
1954; Bran et al, 1959; Hitchner and Johnson, 1948; Hitchner, 1950; Hitchner et al., 1950) to 15 per cent for one or two 1961; van 1959, (Hoekstra, weeks Waveren, 1955) and 20 to 50 per cent for two to four weeks (Doll et al., 1950a). Duration of immunity. Determination of the degree and duration of the im munity engendered by the Bl virus has presented difficulties, as already outlined on page 89. This has hindered the estab
of uniform vaccination proce for different programmes of husbandry and degrees of
lishment
and
dures
(Anon., 1962a; Surin, 1959). Clinical effects of vaccination. In young chicks, clinical effects produced by the Bl
methods
strain have depended largely on the route or method of vaccination. Thus, adminis
immunity
tration by intranasal
or conjunctival drop
or in drinking water has usually had little or no clinical effect (Hitchner and John son, 1948; Luginbuhl et al., 1955; Lulic and Spalatin, 1956; Hutson, 1953; Markham et al., 1951; Miyamoto and Nagashima, 1957; Raggi and Lee, 1962; Russeff and Miteff, 1957; Winterfield et al., 1957), but some instances of more marked clini cal effect have been reported
Administration as 1950a). aerosol has produced either
et al.,
(Doll a
dust
or
no adverse
reaction (Crawley and Fahey, 1 954; John son and Gross, 1951; Markham et al., 1955a);
a slight respiratory
ing a few
reaction
last
(Hitchner and Reising, 1952, 1953b; Price et al., 1955; White et al., 1954) ; or a severe respiratory reac tion with appreciable mortality (Bankowski
and
days
Hill,
vaccinated
1954), especially if the chicks were exposed to E. coli
(Gross, 1961a) or Mycoplasma (Bankowof young 1961b). The vaccination chicks in incubators or in chick boxes has ski,
given variable results (Johnson and Gross, 1952), and the spray method has been considered
unsuitable
for
baby
chicks
exposure.
A
number of reports on the duration of initial vaccination of after
young chicks are summarized in Table 14. The data in this table are confined to observations made two
based on challenge exposure months after vacci
or more
nation. Results
of
tests
conducted
approxi
vaccination of young chicks have been reported by Doll et al. (1950a, b, c) , Hitchner and Johnson (1948), Hitchner and Reising (1953a), Hoekstra (1961), Johnson and Gross (1951, 1952), Jungherr and Markham (1962), Markham et al. (1950), Miya (1957), Pette and Nagashima moto (1961), Rao (1955), Richey and Schmittle (1962), White et al. (1954) and Winterfield et al. (1957). Relatively fewer reports describe the immunity resulting from vaccination at 8 weeks of age and older. Vaccination of such chickens has resulted in satis factory immunity for 12 weeks (Bosgra 20 weeks and Roerink, 1961), (van mately
one
month
after
1955), 26 weeks (Bran et al., 1957), 1959; Miyamoto and Nagashima, 32 weeks (Raggi and Lee, 1960) and 56 weeks (Bankowski et al., 1957; Raggi and
Waveren,
with
Lee, 1962). in the The problem of revaccination immunity residual absence of or presence
B 1 virus has caused a drop in egg produc (Bankier, tion ranging from negligible
may be summarized as follows. The intra nasal B 1 vaccination of passively immune
(Hitchner and Reising, 1953b). In susceptible pullets, vaccination
98
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chicks
1 week
than
less
in appreciable neutralization
of age results of the vaccine
et al, 1960; Brandly, (Lancaster is lessened by neutralization 1955) . This administering the vaccine via the respira
Strain F The suitability
strain
of this lentogenic
by
virus
as a vaccine
tory route which
(1952) of Weybridge, England. Reports on its use in Europe, Asia, Africa and America have also been reviewed
infection
Hitchner
route of
is the natural
and
(Burnstein
1958;
Bang,
al,
1950). Nevertheless, such vaccinated chicks are generally susceptible when
et
of
1 month
this time
has given
response
and
anamnestic
good
a
at
a
immunity satisfactory virus. By way of contrast,
against virulent the similar
Revaccination
age.
of young chicks
vaccination
carrying no maternal
antibody
sulted in a satisfactory
immune
has
re
response.
four or more weeks later, in the presence of antibody, has resulted Revaccination in
a
anamnestic
poor
al,
caster et
1960),
been found
(Lan
response
and such chicks have
susceptible
to virulent
virus
status of known with
later. The immune
two months commercial
chicks
is seldom
and this increases the difficulty
certainty
of establishing a vaccination programme suitable for all situations. For this reason, the age at which primary and subsequent are conducted
vaccinations
must be care
fully evaluated for each particular
set
of
circumstances. When
the
Bl
strain has been used for
both the initial vaccination ations,
considerable
cedure
have
factory
immunity.
vaccinated
at
given 1
been revaccinated
and revaccinin
differences adequate
and
pro satis
Thus, chicks initially to 7 days of age have at 4 weeks
(Hitchner
1952; Johnson, 1956) or 19 (Hitchner et al., 1951b; Johnson, 1956) . Chicks initially vaccinated at about 2 weeks of age have been revaccinated
and Reising, weeks
(Dardiri and Yates, 1962), (Meyn and Pette, 1961), 12
at
4 weeks
8
weeks
weeks
(Hoekstra,
1961)
and
20 weeks
(Hitchner and Reising, 1952). When the initial vaccination has been delayed until 8 weeks, revaccination at
27 weeks
1957) . 100
of
age
has been conducted
(Bankowski
et
al.,
was first reported
virus
Asplin
(Lancaster, resembles
(Anon.,
properties slowly et al.,
F closely Strain 1962b). the Bl strain in many of its
1960) vaccination.
It multiplies
1959).
of all
in chickens
ages
(Quesada and is well suited to their
In
Clinical effects of vaccination. and laying
chickens
hens, Strain
young
F
virus
has sometimes produced mild respiratory symptoms (Asplin, 1952; Asplin et al, 1949). However, egg production has not 1952; affected (Asplin, been seriously Binaghi and Nardelli, 1955; Lancaster, 1957a; Mitev and Gagov, 1960b). No adverse effect on chickens of any age was
(1962), Anon. Nardelli (1955), (1962), Mazzaracchio and Borzemska Orfei (1956), Petek and Gagliardi (1954), or Rao and Agarwal ( 1960) , although a number of different routes of vaccination were used. Contrary findings were report ed by Russeff and Miteff (1956) who observed a slight transient paralysis of the feet, and by Thorne and MacLeod (1960) who found that their strain of this virus reported by Ageeva
(1960),
Binaghi
et al.
and
caused nervous and intestinal symptoms in The cockerels. groups of 4-month-old findings of the last-named authors do not correspond with the evidence at present Strain
characters
on the lentogenic
available
F
of
virus.
Duration of immunity.
A
number
of
of immunity, as determined by exposure to virulent virus, are summarized in Table 15. Other re reports
ports Strain
on the duration
describing
F
virus
immunity have
been
Ageeva et al. (1962), Baldelli
tests
written
with by
(1956), Bor
(1962), Gagliardi and Irsara zemska (1958), Marek (1960), Marek et al. (1961), Mitev and Gagov (1960b), Petek and Gagliardi (1954), Rao and Agarwal
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and
least four months has been reported (Oberfeld, 1962).
Strain
When administered as an aerosol, the LaSota strain has been found effective
(I960), Winterfield et al (1957) Wynohradnyk et al. (1958). on
Studies
with
revaccination
F virus have been of two main kinds. A which has given satisfactory procedure results has been to revaccinate with Strain F virus every one to three months (Mitev 1954). An alternative method has been to administer Strain F virus intranasally to day-old chicks and revaccinate with the
Mukteswar vaccine virus at about 6 weeks of age. This method has proved practical in India (Rao and Agar1962), Malaya and Hong Kong wal, mesogenic
1959).
Other
lentogenic
differences,
Other
1959).
compared
as
strains
vaccine
de
and
(1961) Quiroz Pagnini (1956) have proved satisfactory scribed
Vega
by
in rather
local
situations.
Reports
have
immunity and followed the mortality post vaccination of a live intraocular administration to day-old chicks vaccine attenuated (Gharib, etal, 1961). also
indicated
LaSota Strain The LaSota strain of lentogenic virus differs in some respects from the Bl and F strains. One difference is a lower mean death time for chick embryos (Anon.,
1962).
Other Lentogenic Strains
1960b; Petek and Gagliardi,
and Gagov,
(FAO-OIE,
(Borzemska,
that
poor
Mesogenic Strains Komarov (or Haifa) Strain This strain was isolated in Palestine from an outbreak of Newcastle disease associated with 80 per cent mortality.
with Bl and F, include a greater spread ing potential for LaSota (Marek, 1960)
Serial
and often more post vaccination
1946a) modified the virus to the extent that adult fowls showed no symptom after
respira
tory symptoms (Winterfield et al, 1957). When administered in drinking water, the serological response has been higher with LaSota than with Bl and F strains. As a result,
it
been
has
LaSota strain be used
suggested
at 18 weeks
chickens
to revaccinate
that
given
of
the age
Bl or F
strains at an earlier age (Winterfield et al., 1957).
When
the
immunity
water-administered been challenged 28
engendered
per cent of the chickens
immune
11
by
LaSota vaccine has with virulent virus, only
weeks
after
have been vaccination.
However, two doses of the vaccine at 5 and 28 days of age have resulted in 94 per cent protection
against virulent virus at 15
weeks of age (Dardiri et al., 1957). Im munity has lasted 42 weeks after the sec ond dose of water-administered
vaccine at
8 and 21 weeks of age (Stumpel, 1962). Under field conditions, an immunity of at 102
intracerebral
ducklings
injection posed
to
passage and
(Komaroff
and remained virulent
through
Goldsmidt,
healthy when
virus
18
days
ex
later
(Komarov and Goldsmit, 1946b). Clinical effects of vaccination. In grow ing chickens over 4 weeks of age, the Komarov strain has been well tolerated (Crowther, 1952; Komarov and Gold smit, 1947), although, in young birds in poor condition, a few cases of paralysis have resulted (Thorne and MacLeod, 1960).
Vaccination of laying hens has caused fall in egg production (Golem and 1950; Ileri, 1950; Thorne and Berke, MacLeod, 1960) which has lasted four weeks (Crowther, 1952). Duration of immunity. The Komarov a
strain has been administered
intraocular-
ly (Castro Amaro, 1964), subcutaneously and intramuscularly (Komarov and Gold smit, 1947) with similar results. The
web vaccination method have been given
ly used in Asia (FAO-OIE, 1959; Memo., 1955; Seetharaman, 1951a, b), in parts of Northern Croatia (Lukacevic, 1955), in Bosnia (Marusic, 1955) and in Burma
by Bankowski and Rosenwald
(Peatt, 1945).
stick
wing-web
method
has
also
been
widely used (Madhusudan, 1957; Vandemaele, 1961). Illustrations of the wing-
Martini
Kurjana
and
( 1956) and
(1950);
and
the
Clinical effects of vaccination.
by Schoening
chicks,
and Thompson (1955). Vaccination of growing or mature birds
severe
method
has
has been described
in
resulted
virulent
an
immunity
against
virus
7 weeks lasting (van 1955), 3 months (Komarov and Goldsmit, 1947; van Waveren and Zuij-
Waveren,
1953), 8 months Valadao, 1961), 8 to
dam,
1956), and
13
(da Camara and 12 months (Ileri,
9 months (Komarov et al., 1948a) months (Karrar and Mustafa,
1964; Thorne and MacLeod,
1960).
Serial passage of the Komarov strain in bovine kidney tissue culture has at the
tenuated
to
neuropathogenicity
the
level of the lentogenic strains of New castle disease virus, without loss of anti
(Huygelen
genicity
1963). In comparative and
genicity
tests
and
Peetermans,
based on patho
antigenicity,
the Komarov
to the Mukte(Komarov and Goldsmit, 1947), a formolized vaccine (da Camara and Valadao, 1961), Strain F (Thorne and MacLeod, 1960) and the Roakin strain (Ileri, 1950). strain has been preferred
swar
strain
Mukteswar Strain Reviews of the early work on the devel of a live vaccine in India have been given by Haddow and Idnani ( 1 946) , Iyer (1943), Iyer and Hashmi (1945) and Seetharaman (1951b). It was found opment
that after bryos,
"R..B",
repeated passage in chick
an
attenuated
was
obtained
strain,
em
designated
(Haddow
and
1946). From these studies the vaccine virus, now usually designated the Mukteswar strain, was developed (Dhanda Idnani,
et
al.,
Memo.,
1958;
Gupta and Rao,
1959b;
1955). This strain has been wide
and
the R2B reaction;
Rao,
1959b;
virus
In young
has produced
a
(Gupta Haddow and Idnani,
and
mortality
may reach 30 per cent (Rao and Agarwal, 1960). It has caused paralysis (Daubney and Mansi, 1948) in about 2 1946)
per cent of birds
(Memo., 1955). In chickens over 6 weeks of age, the Mukteswar strain has been well tolerated (Dhanda et al., 1958; Haddow and Idnani, 1946), although mortality varying from 1.3 per cent (Generoso and Mendoza, 1950) to 6 per cent (Haddow and Idnani, 1946) and 16 per cent (van Waveren and Zuijdam, Nervous 1953) has occurred. symptoms have also followed vaccination
(Jaksic and Stefanovic, 1957). As with other mesogenic viruses, the Mukteswar strain causes a marked reduc tion in egg production lasting one to three weeks (Haddow and Idnani, 1946; Ileri, 1956), or as much as six weeks (Memo., 1955). During this period, egg production may decrease 10 to 16 per cent (Agcanas and Rigor, 1 95 1 ) or even 60 per cent
(Dixit, 1950). One of the Mukteswar stocks of vaccine virus has shown increased pathogenicity to White Leghorn chickens causing 56 per cent mortality, as compared with 4 per cent mortality in Rhode Island Reds. This selective pathogenicity has been attributed to repeated passage through White Leg horn embryos (Nandi, 1955).
Duration of immunity. Using the sub route, Haddow and Idnani (1946) found immunity against virulent virus to be durable for 9 to 15 months. Similar results have been reported by Cakalowa et al. (1955) and Daubney and Mansi (1948). Immunity lasting three to four years was noted by Nilakantan et al. (1960b) and Seetharaman (1951a). In cutaneous
103
contrast,
(1949) were of
et al.
Bornstein
the opinion that the immunity
produced
by the Mukteswar strain was durable no more
than
one year.
Significant
for
HI
of doses of mesogenic used in the United
that the number
vaccines
wing-web
States has declined appreciably since 1953 (Anon., 1962b). Nevertheless, mesogenic strains of American origin have
titres have been present in 11 to 64 per
vaccine
cent of fowls vaccinated
of countries in (Kaschula, 1950; Vandemaele, 1961) and Europe (van Waveren, 1955). Clinical results of vaccination. In birds under 6 weeks of age the Roakin and MK107 vaccines can cause severe mor bidity and some mortality (Anon., 1962b; Cole and Hutt, 1961; Van Roekel, 1956;
6 to 12 months
(Lukacevic, 1955).
previously
Africa
Using the wing-web stick and intramus cular methods of vaccination Dhanda et (1958), Generoso al. and Mendoza (1950) and van Waveren and Zuijdam (1953) found that immunity lasted 6 to 8 months
but declined at 12 months
(van 1955). A similar 8 months' has resulted from a number of
Waveren, duration different et
al,
vaccination
routes (Nilakantan including administration
1960a),
in drinking water (Forsek et al, 1957). Nilakantan et al. (1960a) have concluded that there is little difference between
antigenically
Mukteswar and Komarov
the
been used in a number
strains.
week-old
(Clancy
tion
Using electrophoresis of serum samples, et al.
et al.,
vaccination
significant
1949). A lower but mortality has been
of two (Cole and Hutt, similar strain resis
related to the genetic characteristics strains of White Leghorns
of
196 1 ) . Evidence
a
disease virus
tance to Newcastle strain)
Lukacevic
1955). Thus, 50 per cent of chicks have died after vaccina
van Waveren,
(Roakin
has been reported by Francis et al.
(1964). However, Beaudette et al. (1949) Cordier-Boullangier et al. (1955b) found that, with the Roakin virus, mortal
(1958) showed that vac cination with the Mukteswar virus resulted
and
in a significant increase in gamma globulin
ity was negligible in chickens vaccinated
which reached a similar level to that
5 weeks
tained in experimentally
infected
at
fowls.
of
age.
In 4-week-old
Roakin virus has resulted in
chicks,
at the
a significant
decrease in weight gain following vaccina
Strains originating in the United States Two mesogenic strains of the virus have used in the United States as
been widely
tion (Francis et al., 1964). With the MK107 virus,
mortality
has
also been negligible in 4-week-old chicks (Markham et al., 1949). A differential characteristic between the Roakin and
MK107
viruses
strain (Roakin) was identified during the screening of 105 strains (Beaudette et al.,
Rosenwald
(1959) who found that the Roakin strain consistently
1949). The other strain (MK107) subjected to serial passage in chick
was
failed
and
MK107 just
live wing-web
duck
administered
embryos
(Clancy
et
1949). Reports velopment
al.,
vaccines. One
following its isolation Markham et al.,
1949;
of wing-web
vaccines
in the
States
include
(1949c,
1951),
(1948b), Van (1948) and Van Roekel
et
al.
those
by
Beach
Beaudette
(1956). It would appear from published figures 104
to
inoculated
been
reported
by
spread
to
contact
as consistently
birds, spread
the
from
birds to contact birds in other
cages in the same unit.
A and reviews relating to the de
United Roekel
whereas
has
et al.
number
of reports have shown
that
are well tolerated by birds 4 to 16 weeks of age (Anon., 1962a; wing-web
vaccines
Quinn and Thompson, 1952; Thompson 1956), and Osteen, 1952; Van Roekel, even when chickens on infected farms are vaccinated
al,
(Beach,
1949c; Beaudette
1949; Markham et
al,
et
1949). How
ever, in some vaccinated systemic
reaction,
flocks,
respiratory
a slight
symptoms
or, more rarely, paresis may be observed (Beaudette et al, 1949; Van Roekel,
1956). These reactions (Davis
severe in winter
In
susceptible
have been most et
al,
1950). with hens, vaccination
wing-web vaccine viruses has reduced egg production and egg quality to almost the same extent as natural outbreaks of New (Beach,
1949c;
Beaudette
castle
disease
et
1949; Kaschula, 1950; Van Roekel
al,
1956; van 1948; Van Roekel, et al., Waveren, 1955). Duration of immunity. Wing-web vac cines have produced
an adequate and per
immunity (Cordier-Boullangier et Richter, 1953; Van Roekel al., 1948; Van Roekel, 1956; Thompson
sistent
al, et
and
1955b;
Osteen,
1952),
sufficient
to protect
against a drop in egg production for one laying season (Bankowski and Rosenwald,
1956). The immunity engendered by the Roakin strain has been durable for 12 months (van Waveren, 1955; van Waveren and Zuijdam, 1953). The MK107 strain has resulted in immunity against virulent virus lasting 10 weeks (Markham et al., 1954) to 4 months (Clancy et al, 1949). A feature of many wing-web mesogenic
vaccines has been the ability of the vaccine
virus to spread to adjacent flocks (Beach 1949c). Kaschula (1950) was of the opin ion that there was no danger of the Roakin strain becoming virulent. Although the mesogenic ministered muscular
vaccine viruses are usually
ad
either by the wing-web or intra routes, Kaschula (1952b) found
for revaccination with the Roakin virus, intraocular instillation was the best that,
method.
and reviewed
by Iyer (1943) and Kran-
eveld (1950). A strain of virus originally isolated from an outbreak of Newcastle disease in Hertfordshire, England, was used. After 14 to 33 passages in the chick embryo, the virus did not kill adult fowls; and vaccinated chickens survived exposure to virulent virus (Iyer and Dobson,
1940). Other studies with the Hertfordshire vac cine virus have also been made.
Clinical effects of vaccination. In chicks possessing passive immunity, the Hertford shire strain has caused no mortality and Beke, 1955). However, (Szakmary chicks under 8 weeks of age have been very susceptible (Iyer and Dobson, 1940; Schneider, 1954). The vaccine has proved satisfactory for chickens over 12 weeks
old
1954); although some of paralysis have followed vaccina tion (Mazzaracchio and Orfei, 1955; Salyi and Hodosy, 1952), and there has been a severe drop in egg production (Pagnini, 1954). Clinical effects have been less severe when an adsorbed but living Hertfordshire vaccine has been used (Schneider,
cases
(Schmidt,
1952).
Duration of immunity. The intramuscu lar or subcutaneous vaccination of chickens over 3 weeks of age and of adults has resulted in a durable immunity lasting three to five months (Gualandi, 1951; Mazzaracchio and Orfei, 1954; Szakmary and Beke 1955). Thereafter, during the succeeding nine months, the immunity has progressively declined (Mazzaracchio and Orfei, 1955), although Teklinska (1951b) found vaccinated fowls were protected for one year. Some chickens
vaccinated with the Hertfordshire virus have been immune to experimental infection HI although titres were considered
Hertfordshire (or Herts) Strain Initial studies on the attenuation of a strain of Newcastle disease virus for use as a live immunizing agent were conducted by Iyer and Dobson (1940)
virulent
negative (Hamann,
1958).
Gualandi (1950) concluded that chick with live Hertfordshire vac
ens vaccinated
cine did not excrete the virus not a source of infection, contrasts
with
other
and were
a finding
results
that
(Lancaster,
1963a). 105
In some areas of Hungary, a strictly vac controlled nation-wide compulsory cination with the Hertfordshire strain of virus, together with veterinary and police control to ensure 100 per cent vaccination coverage, resulted in a marked decrease in the incidence of Newcastle disease (Zsigmond and Gyorgy, 1957).
authors considered that virus administered by the vent was less likely to be neutralized vaccination.
Further reference to mesogenic is made
under the heading
ference"
on page 93.
mesogenic
strains of Newcastle
disease virus
have been
developed
examined
immunizing
agents.
as
and Some
have been attenuated by passage through mammals (Reagan et al, 1947b; 1948c, d, 1952c) . Others have been attenuated by passage through duck and chick embryos
(Papparella, 1956; Prier, 1951; Collier and Dinger, 1950); young ducklings (Pillai, doves 1949); striped ground {Geopelia striata) (Mansjoer, 1961; Mar tini and Kurjana, 1950) or tissue cultures (Rusev, 1960; Russeff, 1960). Moreover, some mesogenic strains have been found suitable for use in Indonesia (DonkerVoet and Kurjana, 1950; Mansjoer, 1961) and in certain countries of Africa (Jezierski, 1953) and of South America (Nobrega, 1955). On the other hand, serial passage in chicken embryos has in creased the virulence of a vaccine strain of virus to the point where its use in the field
could
not be recommended
ring, 1958). Mesogenic cine viruses
(Geh-
pathogenic
vac
have been used successfully
for revaccination of chickens initially vac cinated with an inactivated virus (Adler et al, 1951; Brandly et al, 1946c; Dardiri et al, 1961; Kaschula, 1952); and with a lentogenic virus (FAO-OIE, 1959; Rao and Agarwal, 1962; van Waveren, 1955). Winterfield and Hitchner (1961) showed
that
revaccination
with the live strain "N-47" gave
route
by
weakly
106
small-scale
a
better
the
vent
or intramuscular
extract of tobacco mosaic virus into chick ens engendered
al
ger et
to Newcastle
immunity
by San (1961) failed to confirm these
disease. Subsequent
investigation
results.
Tissue Culture Vaccines The use of tissue-culture-propagated vaccine viruses is discussed on page 130.
Inactivated Vaccines This section summarizes the work that has been done up to now on the inactiva-
tion of viruses by different substances and procedures
and the results obtained.
Inactivation by
Beta-Propiolactone of viruses
inactivation
(BPL)
by beta-propiolactone
was first re
ported by LoGrippo and Hartman (1955), and this method was also used by Mack and Chotisen inactivated general
BPL
(1955,
Newcastle
review
1956) to prepare an disease vaccine. A
of virus
inactivation by (1960).
has been given by LoGrippo
He showed that, with rabies virus, the use of BPL and ultraviolet irradiation in com bination resulted in increased viricidal ac tion, with the antigenic
serological
better preserved than when
routes. These
by Marxer
experiment
indicated that injection of an
pathogenic
response than did the Roakin strain by the wing-web
A
et al. ( 1958)
The satisfactory or weakly
strains
"Viral Inter
Extract of Tobacco Mosaic Virus
Other Mesogenic Strains Other
from the initial
by circulating antibodies
alone. The inactivation ease
virus
at
component
BPL
being
was used
of Newcastle dis 37°C has been attained by
TABLE 16 — Duration of Immunity following Initial Vaccination of Chickens with Beta-propiolactone Inactivated Vaccine (Lancaster, 1964a) Author
Concentration of B.P.L.
Age when vaccinated
Mack and Chotisen, 1 955, 1 956 Winmill and Weddell, 1961 Simmins and Baldwin, 1963 Haig et al., 1962 Sullivan et al., 1 958 Gill era/., 1959 Keeble and Wade, 1963 Keeble and Wade, 1 963 Keeble and Coid, 1 962 Piercy et al., 1 962 Appleton et al., 1 963 Christie ef al., 1 963 Akat, 1 962 Cooper, 1 963
0.025% 0.025% 0.025% 0.033%
4 weeks 2 weeks
*
2 weeks
0.1% 0.1%
3 weeks 1 0-1 9 days 2 days
0.1% 0.1% •
3 weeks 2 weeks
0.2% •
11-25 days 1-14 days
m m
adult
(a) a concentration of BPL of 1/1,000 to 1/2,000 for 30 minutes; (b) a concentra tion of 1/5,000 for 60 minutes; or (c) a concentration of 1/7,000 for more than two hours (Cherby and Valette,
In field trials,
virus
a
1964). inactivated by
has revealed a potency similar to that
of a live vaccine
(Gill
and Stone, 1960).
The relative value of formalin and betaas inactivating propiolactone agents for disease
Newcastle
virus
has
been
dis
cussed by Piercy et al.
Appleton
et
(1963) and by (1964). The former
al.
authors believed that prolonged of virus to
BPL
adversely
affect
exposure
at low temperatures
the
antigenicity
might
of the
Appleton et al. (1964) felt that more study with BPL was needed because birds vaccinated with BPL-inactivated virus had shown a poor immunity against challenge. The addition of adjuvants to BPL-inactivated Newcastle disease virus vaccine.
has
been
examined
by
Gill
al.
and
Sullivan
et
al.
These
(1958) (1959). studies have been extended by several other workers, as shown in Table 16, and most recently
8 weeks 2 weeks
0.1%
at least 1 6 days at least 3 months at least 8 months at least 8 months
approximately 2!4 months approximately 3 months at least 13 days at least 10 weeks at least 1 2 weeks at least 1 8% weeks
poor at least 6 weeks from 2 to 6 months at least 1 3 weeks
to writer of this Review
Details not available
BPL
adult
Duration of immunity against virulent virus
et
by Hofstad et al. (1963).
Clinical effects of vaccination. chicks
without
been vaccinated
Day-old
maternal
antibody have (Haig et al., 1962) with
retardation of growth subsequent (Keeble and Coid, 1962). The vaccination of laying flocks with a commercial BPL-inactivated vaccine has out
had no significant effect on egg produc tion; nor any respiratory symptoms
(Cooper, 1963). Duration of Immunity. Results of a number of studies are summarized in Table 16. It was shown by Simmins and Baldwin (1963) and Piercy et al. (1962) that Strain
F
virus (described under the heading "Lentogenic Strains") produced as good a
BPL-inactivated vaccine as the more viru lent Herts strain. Thus, should inactivation of the virus be incomplete, there would be no ill effect among vaccinated birds if Strain F virus were used. The presence of active virus residue in a
BPL
vaccine was
Appleton et al. (1963). Pini et al. (1963) found in activated virulent virus to be more im
considered
possible
munogenic
than Strain
by
F
virus. 107
IMMUNE RESPONSE
VACCINATION WITH A
FOLLOWING
BPL VACCINE 1280
I
I
I
I
I
I
I
1
2
4
6
8
10
12
14
16
Interval
after vaccination (weeks) (Redrawn
from Keeble and Wade, 1963)
Figure 18.
Of the results summarized in Table 16 poor immune response re ported by Appleton et al. (1963) should Under the condi be noted particularly. tions of their experiment, these authors found that inactivation with formalin resulted in a better vaccine than did BPL. Appleton et al. (1964) have discussed the the generally
variables evaluation
involved
in the preparation
of inactive Newcastle
and
disease
vaccines and have suggested that the bal ance between the concentration
fluid needs further Appleton et al. (1963) used inactivating temperature of 4°C for 48
and
amnio-allantoic
investigation. an
of BPL
108
hours; whereas Haig et al. (1962), Mack
(1955), Keeble and Wade (1963), Simmins and Baldwin (1963), Winmill and Weddell (1961) used 37°C for two hours; and Gill et al. (1959) and Sullivan et al. (1958) used 21°C for two and Chotisen
hours. Vaccination
programmes
utilizing BPL-
vaccines. In some studies (Sul livan et al., 1958), initial vaccination was inactivated
at 2 weeks
of
6 weeks
of
cination
at 2
age;
revaccination
was at
More recently initial vac to 3 weeks of age has been
age.
followed by revaccination at 12 to 20 weeks of age. Such procedures have given
o
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109
a marked secondary response, as shown in Figure 18 (Keeble and Wade, 1963), and
To increase the immune response, dif ferent adjuvants and adsorbents have been
al,
Gill
of immunity after the second of 17 weeks (Simmins and vaccination Baldwin, 1963), 20 weeks (Gill et al, 1959) and approximately 52 weeks (Gar-
tested (Brandly et
side, 1962).
increase the immunizing properties vaccine (Nakamura et al, 1956).
a duration
In England and Wales, a modification of the above schemes has been to recom at intervals before
mend three vaccinations
the pullets come into lay (Anon., 1963a; Smith, 1963), and another to conduct the initial vaccination when the chicks are 10 days of age (Hemsley, 1963). However, Keeble et al. (1963) considered it best, in the case of chicks
from well-immunized
stock, to delay the initial vaccin
parent
ation until the chicks
of
published
gel has been
been
has created
probably
with
a
BPL
vaccine
by Formalin
Nakamura vaccine,
reviewed by Hofstad et
al (1963). Viru been
have generally
(Surin, 1959),
con
it being
sidered that these strains produce strains.
better
agents than mild or lentoHowever, Hanson et al.
(1951b) have shown that this is not neces sarily the case.
Because strain differences
in immunogenic considered
potency exist, it has been
advisable to use several strains
in the production
of
a vaccine
(Brandly
1946b). Formolized virus has been administered by the subcutaneous, intra
et al.,
muscular,
and intraperitoneal routes (Beach, 1945; Brandly et al, 1946b; Nakamura et al, 1937, 1956). Treatment intravenous
with formalin has been found more effec tive than other methods of inactivation Acevedo et al. 110
condi
al,
et
al,
prepared
by formalin-inactivated
genic
favourable
for the steady antigenic response (Rao, 1955). In the absence of an adju vant, formolized vaccine has been more tions
and the overall results obtained have been
immunological
adju
(Generoso and Agustin, 1947; Jacotot and Vallee, 1959; Skoda and Zuffa, 1957). The adjuvant
et
so treated
with different
noted
Formalin has been used extensively for the inactivation of Newcastle disease virus
lent strains of virus
to
of the
differences in the immune
associated
have
shown
1956). A formolized from embryonic fluids only, has been found to be better than one from fluids, membranes prepared and 1 962). The use of embryos (Perez-Rebelo, a minimum concentration of formalin is also a factor for consideration (Appleton
disease on
(Table 17).
Inactivation
vants
al,
(1963)
by Simpson
in unvaccinated flocks and
in flocks vaccinated
Considerable response
et
effective when administered intravenously (Brandly et al, 1946b; Madison, 1947;
show the effect of Newcastle egg production
ium hydroxide
1946b;
The adsorption of virus with alumin
are about 3 weeks
age.
Figures
1959; Traub, 1943b). a formalin-inactivated
by
and Mendoza (1947), Appleton
(1963) and Coronel (1947).
1963).
The evaluation
of immunity
produced
vaccines has been
complicated by the use of different routes of inoculation and various strains and doses of challenge virus (Appleton et al, 1963; Hofstad, 1953b, 1956). Although survival
after challenge
immunity
accurately
may not evaluate
(Hofstad,
1953b),
this criterion has been generally adopted. Clinical effects of vaccination. Formalininactivated vaccines have been adminis tered to chicks at 1 to 2 days of age with few, if any, clinical side effects (Rao, 1955, 1956; Chute, 1952; Hofstad, 1954b;
Waller and Gardiner, 1952). Under field conditions and with chickens of varying ages, the vaccine has been considered safe (Atanasiu and Gareau, 1951; Haddow and Idnani, 1941; Jacotot and Vallée, 1959). Egg production has not been affected
(Placidi and San-
tucci,
where
1953c),
except
incomplete
TABLE 18 — Duration of Immunity Following Initial Vaccination of Chickens with Formalin Inactivated Virus (Lancaster, 1964a) Age when vaccinated
Author
% surviving challenge1 Months after vaccination
2
Chute, 1952 Waller and Gardiner, 19533 Rao, 1956 Miyamoto and Nagashima, 1957 Dardiri era/., 1957 Waller and Gardiner, 19533 Hanson et al., 1951b Hofstad, 1 953a4 Jacotot and Valine, 1 962 Rao, 1 956 Mitchell and Walker, 1952 Jezierski, 1 953 Hofstad, 1 953a4 Nakamura et al., 1956 Mitchell and Walker, 1951a Mitchell and Walker, 1953 Mitchell et al., 1952 Miyamoto and Nagashima, 1957 Traub, 1 944
1 day 1 day 2 days 4 days 10 days 10 days
3
4
0
0
72 100
66
11
9
12
02 62
30 100
48 80
98 84
96
3 weeks
88
80
S
3 wscks
82
3 weeks 6 weeks 8 weeks
young 3 months 4 months
7
100
100 100
100
100
adult
90
s
S
100
100
75
66
100
87
80
90 90 60
adult adult adult adult
1 In many reports the end point of the duration of immunity was not determined 2 No immunity compared with non-vaccinated controls 3 Data taken from Fabricant (1956) 4 Results with smaller challenge dose S — Immunity considered satisfactory
or dissociation
of the virulent
chickens
virus in the vaccine has resulted in frank
marized
inactivation disease
(Mitchell
et
al.,
1956b). One criticism against
Placidi,
the addition
been the persistence
tissue
of has
of the adjuvant in the
(Schoening
and
Osteen,
1948). In addition, certain oils, when used as adjuvants, have caused some post reaction (Generoso and Agustin, of the granuloma type (lacotot and Vallée, 1962). However, the use of an
vaccinal 1947)
aluminium
hydroxide
gel
has not caused
under field conditions problems (Eskelund, personal communication). Duration of immunity. The immunity
any
that
follows
the
initial vaccination
of
of age is sum This summary is of virulent virus for
1
day 18.
based on the use challenging
to the vaccines for broilers
adjuvants muscle
1952;
over
in Table the
resulting
immunity.
Authors who have described duration of tests for chickens under 1 immunity Appleton et al. month of age include: (1963); Doll et al. (1951b); Generoso and Agustin (1947); Grillo Torrado and Smitsaart (1962); Hofstad (1953a); Lucam (1949a); Perez-Rebelo (1962); Rao (1955); Schneider (1956); Traub (1943a) and van Waveren
A
(1955). number of authors
favourable
have
reported
results after the use of formo-
lized vaccine. In some instances the results in were not determined by challenge; others serological
procedures
were used.
Ill
authors
These
include:
and
Acevedo
Mendoza (1947), Atanasiu and Gareau (1951), Balducci et al. (1954), Beach (1945), Botija and Loizelier (1948), da Camara and Valadao (1961), Coronel (1947), Dedie and Starke (1952), Ellis and Crook (1953), Freudenberg (1950), Haddow and Idnani (1941), Kaschula (1950, 1952b), Lucam (1949a, b), Mansjoer (1961), Netter and Nguyen-ba-Luong (1956), Pagnini (1950), Placidi and Santucci
and
(1953c)
months fowls
depending
In duced
In contrast, a number of authors have had poor results after the use of formolized vaccines. These include: Adler et al.
days;
et al. ( 1949),
Iyer (1943), Gualandi (1950), Levine and Fabricant (1952), Nakamura et al. (1937), Pomeroy (1951), Sturgess (1931) and Winmill and Weddell (1961). Two methods have been adopted to prolong the immunity from resulting initial vaccination with a formolized vaccine: revaccination with a formol in activated vaccine,
and revaccination
with
a live vaccine.
the
used (Jacotot
on and
weeks;
10VS
which
programme
mortality
large
a
at
re
farm,
15 to 17
to 7 months
6
(Ellis and Crook, 1953). The value of using a live vaccine for revaccination after a formolized vaccine has been examined. Thus, in the initial programme, of a vaccination stages formolized vaccine was used for laying stock and followed by a live vaccine two or three weeks later (Adler et al, 1951). A similar programme has been suggested by Le Dosseur and Lissot (1949). How ever,
a
longer interval has been by Kaschula (1952b), who that at least 12 weeks, pre
much
considered
disease vaccine depends to a great extent on the interval between doses (Hofstad, 1954b; Woernle and Brunner, 1957). A
a
second dose of vaccine
2 weeks after the
has not
appreciably improved im munity (Doll etal, 1951b; Hofstad 1953b; van Waveren, 1955), whereas a second weeks
strong
1959).
formolized vaccine was used
ferably
1956) or 8 to 12 weeks later has resulted in a sub stantial degree of immunity (Hofstad, 1954b; Miyamoto et al.. 1957; Rao, 1956; 3
a
recommended
The degree of immunity following two of formol-inactivated Newcastle
dose
in
on the adjuvant
a vaccination
doses
first
resulted
(Nakamura et al., 1956). When aged 15 to 18 months have been
and Vallée,
Schjerning-Thiesen
Clancy
has
vaccinated twice at an interval of 12 days, the immunity has lasted 8 to 12 months,
(1951).
( 195 1), Beach ( 1945),
later
weeks
immunity against virulent virus (Hofstad, 1957; 1956; Miyamoto and Nagashima, Miyamoto et al., 1957) lasting six months (Miyamoto and Nagashima, 1957) to nine
(Schneider,
five months, should elapse before vaccine is used after wing-web
live
with
vaccination
initial
a
formolized
vaccine. When Dardiri and Yates (1962) ad ministered formolized vaccine to chickens at 2 weeks of age, followed by revacci nation with age,
Bl
virus at 4 to 16 weeks of
satisfactory
immunity
lasted
14
months after the last vaccination. Hilbrich and Reuss (1963) studied a reverse pro cedure in which the initial vaccinations of Bl virus, followed
Waller and Gardiner, 1952) lasting at least eight months after the last dose of vaccine
young chicks were with
(Hofstad,
adsorbed vaccine. The resulting immunity
1953b). When the initial dose is administered at 3 weeks of age or less, revaccination
should
be
delayed
for
at
at 2 to 6 months
lasted at least
of
age by a
15 months.
The comparative efficiency of formalininactivated vaccines and live virus vac cines has been studied. The formolized
least four to eight weeks (Dardiri et al., 1961; Hofstad, 1953a; Rao, 1956). When the first dose is administered at 6 weeks
vaccine was considered
of
the Komarov vaccine
age or older, revaccination
112
16 or more
formolized
less effective
than
by da Camara and
Valadao (1961) and less effective than the Mukteswar vaccine by Zuijdam (1952a). Netter and Nguyen-ba-Luong (1956) con cluded there was little difference the
Mukteswar
virus
and
a
between
formalin-
1930; Nakamura et al., 1937; Topacio and Coronel, 1939), even though adjuvants were added (Fortner et al., 1959). In con trast, inactivation with the sodium salt of 8 - hydroxy - 7 - iodoquinoline - 5 - sulphonic
adsorbed vaccine. In contrast, when com
acid has produced
pared with the lentogenic vaccine viruses, Bl and LaSota, a formolized vaccine has
(Kraneveld and Nasoetion, 1948). Also, an inactivated vaccine involving 50 per
sometimes
on
cent glycerin
or there (Miyamoto and
administered
better
given
et al.,
(Dardiri
challenge
has been little
Nagashima,
difference
protection 1957),
this
chemical
were
In contrast,
irregular
and
lasting
12 months
(Doyle and
Wright, 1950), especially when a booster injection was given three or four weeks after the initial dose (Legenhausen and Sinkiewicz,
1959);
Legenhausen
et al.,
1959). Two doses of the vaccine have given protection against death 61 weeks after vaccination (Bankowski and Corst1960). On the other hand, crystal violet vaccines have given less protection when compared with either live wing-web vet,
vaccines van
(Thompson and Osteen,
Waveren
and
Zuijdam,
1953;
1952; van
Waveren,
1955) or a tissue culture vaccine (Bankowski and Corstvet, 1960; Bankow ski et al., 1958b). They have also been
found difficult to prepare chell and Walker, 1951a).
Inactivation
in bulk
(Mit
by Other Chemicals
A number of
reports
of 0.5 g/ml, urefound to inactivate a
been
disease virus, as deter
by the absence of chick embryo
Administration of the treated
infectivity.
virus by the wing-web same
HI
route has given the
titres as those resulting from the
use of a commercial
Newcastle
wing-web
(Bower and Eisenstark,
disease vaccine 1954).
have described
Inactivation by Heat Dutcher et al. (1960) observed that for a very short period of time inactivated Newcastle disease virus with a minimum destruction of antigenic proteins. A series of three intramuscular vaccinations at intervals of three to seven high temperature
days resulted
Inactivation Irradiation
immunity
by Ultraviolet
The practical application of ultraviolet irradiation in the inactivation of viruses for the preparation of vaccines has been reviewed by Taylor (1960). Prolonged irradiation has killed the virus with an loss of antigenic associated property (Sinha and Datta, 1950b). The infective property of Newcastle disease virus has been destroyed more readily by ultraviolet light
by a variety of chemicals (Coronel, 1939; 1955; Farinas, Bindrich and Schmidt,
the less
unsatisfactory
in a satisfactory
lasting five to seven months.
results following the inactivation of Newcastle disease virus generally
(Teklinska,
a crystal violet
ethylene glycol vaccine has engendered an immunity
has
mined
The use of crystal violet for the in of Newcastle disease virus has given variable results. Thus, Iyer and Dobson (1941b), Iyer (1943) and Winmill and Weddell ( 1961 ) found the results with
intradermally
strain of Newcastle
by Crystal Violet
activation
unsatisfactory.
has given good results when
a concentration
thane
Inactivation
vaccine
1951).
At
1957).
a satisfactory
than
activity
the
less
sensitive
(Prince and Ginsberg, sensitive
agglutination
(Atanasiu and Suoto Patuleia,
cytotoxic 1957) or property 1952). The 113
phenomenon
of multiplicity reactivation
of Newcastle
disease virus after exposure rays has either not been
to ultraviolet
detected (Barry, 1962) or has been very weak (Drake, 1962). Other studies on the inactivation of the virus by ultraviolet
irradiation have been reported by Drake and Lay (1962). vaccine, utilizing ultra Oil-emulsified violet irradiated
virus, when administered
at 10 days and again at 42 days
given protection the
second
for
of
age, has
at least 20 weeks after
vaccination
(Legenhausen im
Such durable
and Sinkiewicz,
1959). munity was not obtained by Brandly et al.
(1946b).
eggs, in embryonating fornia 11914) the found Eisenstark Nadel and (1955a) yet to allantoic fluid to be non-infective, have a positive HI titre. This fluid was
used
vaccine
a
as
Ultrasonic
wave
treatment
was
em
ployed by Michelsen (1951) and resulted in no appreciable reduction in the infec
tion titre of Newcastle
disease virus.
and 21
that was the same as that engendered by intraocular vaccine (Nadel a commercial and Eisenstark, 1955b). Similarly, Rott 1963) have reported the and identification of the incom plete form and the viral microsome in Particles chicken eggs. embryonating
et
(1962,
al.
isolation
been
by Ultrasonic
intra-
days ocularly to day-old later the immunity was challenged with virulent virus. The "incomplete" virus vaccine resulted in a level of immunity
"incomplete
designated
Inactivation Treatment
administered chicks,
demonstrated
in
virus" have also intracytoplasmic
inclusion bodies found in tumor cell cul tures infected with Newcastle disease virus (Adams and Prince, 1957). Chromato graphic
of
a
analysis
has shown the presence haemagglutinating
non-infectious
of Newcastle (Wilson, 1962b).
component
Inactivated by Different Methods Compared
disease
virus
Vaccines
Using three strains and two substrains of Newcastle disease virus, Hofstad et al. (1963) compared the immunogenicity of Newcastle gamma
(a)
disease
as allantoic
mostly
conditions;
virus fluid,
preparations,
by:
inactivated
irradiation under different 0.1 per cent formalin at
(b)
2° to 4°C; (c) 1:1,000 beta-propiolactone for two hours at 37°C. The immunity was challenged with virulent virus five to eight weeks after vaccination.
The overall
sults showed that beta-propiolactone inactivating
agent was superior
re
as an
to gamma
irradiation and formalin in preserving the of Newcastle disease immunogenicity virus.
one stage in the propagation Newcastle disease virus
virulent 114
To facilitate mass vaccination proce dures, studies have been made in which a live Newcastle disease vaccine has been with other virus vaccines and as a single inoculum (BeauThus, a live lentogenic 1949b).
combined
administered dette,
Newcastle infectious
disease vaccine combined with has virus vaccine bronchitis
as a spray or dust been administered (Crawley, 1954b; Markham et al, 1955a, b, 1956b, 1957; Wardsworth and Young, 1955), in drinking water (Hitchner and 1961; Luginbuhl White, 1956; Hoekstra, et al., 1955) or by other means (Jansen
and Richter,
1959).
of combining New bronchitis infectious on the been questioned
The advisability
"Incomplete" Newcastle Disease Vaccine At
Combined Vaccines
castle
disease
and
of the
vaccines has grounds that the combination may prevent the development of satisfactory immunity
(Cali
to either or both diseases (Bankowski and
Rosenwald, 1956; Bankowski et al, 1955; Hitchner and White, 1956). On the other hand,
good
against both
protection
eases can also be conferred
vaccine
administered
(Markham
et
Mesogenic virus
dis
by a combined
by different
routes
al,
1956b). strains of Newcastle
pigeon
vaccines
pox
disease
with fowl or for wing-web or
administration
(CordierBoullangier et al, 1955a; Delpy and Hars, 1953; Dhanda et al, 1958; Jansen and v.d. Vlerk, 1954; Komarov et al, 1948b; Madhusudan, 1957; Prier, 1951; Suhaci et al, 1958; Van Roekel et al, 1948; Verge, 1954; Weidenmuller, 1954) or by the feather follicle route (Richter, 1956). Similarly, the lentogenic Bl strain has been combined with fowl pox virus for wing-web administration. This has result ed in good immunity to fowl pox but insufficient
protection
disease. Better immunity injections
against
Newcastle combined
and Schneider,
disease virus
1963a). also been
with a fowl plague virus
mixed vaccine (Daubney Lucam,
has
and Ishak,
as a
1953;
1949d).
To meet certain situations, it has been
have been combined
intramuscular
web (Bengelsdorff
Newcastle
has followed two
of the vaccine into each wing
found
convenient
vaccines
and by different has
to
two
administer but
simultaneously,
separately,
This procedure
routes.
been used for Newcastle
disease and
fowl or pigeon pox virus vaccines (Bran et al, 1959; Gupta and Rao, 1959b; Hartwigk and Untermann, 1962; Hutson, 1953; Jezierski, 1953; Lancaster, 1957b) and for Newcastle disease and infectious bronchitis
vaccines
(Bankowski
et
al,
1957).
A
viruses
triple
vaccine,
comprising
of Newcastle
bronchitis
disease,
and infectious
has been administered
live
infectious
laryngotracheitis,
by the vent method.
This procedure has resulted in satisfactory immunity
to Newcastle
disease (Winter-
field and Hitchner. 1961).
115
PART IV: Virus Propagation and Vaccine Production VIRUS PROPAGATION METHODS hours after inoculation;
in Eggs
Propagation
and Ferry (1934) found New castle disease virus was highly infective Burnet
for developing chicken embryos
30 to 48 hours
occurred
embryos
: death
of
after
The technique used for inocu of the chorio-allantoic membrane
inoculation. lation
has been described
in detail
by Burnet
(1936). Early studies on the use of the develop ing embryo in the propagation of viruses have been reviewed by Iyer and Dobson (1940). Using 9- to 13-day-old embryo nating
these authors
eggs,
inoculated
a
V filtrate of a 1 per cent saline of liver and spleen from an in fected fowl. Usually, all embryos were dead by the second day. Material for sub Berkefeld
emulsion
sequent
passages was
harvested
on
the
second or third day post inoculation. Several factors relating to the virulence and propagation
of Newcastle disease virus
in embryonating
eggs
were examined
by
Florman (1948), Murchelano and Han son (1960) and Zebrowski (1957). Under circumstances,
certain
a decline
in anti
genicity was recorded (Beach et al, 1948). Liu and Bang (1953) compared the in the chicken
growth
and pathogenesis
embryo
of three strains of virus differing
considerably
in virulence.
reached a maximum fluid 24 hours the allantoic virus brain 116
All
three strains
titre in the allantoic
following inoculation into However, the virulent
sac.
(Strain CGI 79) appeared and liver of the embryo some 20 in
the
and on the death
of the embryo, the amount of virus in tissues approached fluid.
the
that in the allantoic
In contrast, the vaccine virus (Bl appeared in the tissues of the
strain) embryo
24 to 30 hours after allantoic
sac
and the titre of the virus in the
inoculation,
lower than in the Furthermore, the vaccine virus did not kill embryos within four days. One feature of the Bl strain was its ability to cause severe pneumonia with cellular infiltration and inflammatory tissues was appreciably allantoic
fluid.
exudate in embryos
six days after inocu
lation. It appeared that the lungs of 16- to embryos were particularly sus
1 7-day-old
by the Bl strain. Soon after the development of pneumonia, the embryos began to die (Liu and Bang, ceptible
to destruction
1953). Other reports have indicated that the infectivity of Newcastle disease virus reaches a maximum
of
inoculated
Gareau,
shortly after the death
embryos
(Atanasiu
and
1952).
Certain chicken embryos, when exposed to large doses of Newcastle disease virus, have shown infection
a state
(Karrar,
of refractivity to 1963).
It
is
the
possible
state is indicative of toxicity (Burnet, 1955; Westwood, 1961). Although some embryos have died within four days, no haemagglutinin has been detected (Karrar, 1963).
that this refractory viral
A
number
of substances have tended
to protect chick embryos against infection with
Newcastle
disease virus.
These
in
clude Trypan blue and related compounds 1961a) and polylysine (Green 1953). Furthermore, when a cul
(Finkelstein, et al.,
ture of E. coli or its endotoxin inoculated
into
allantoic
of
sac
embryos some hours before
embryonating the
the
has been
resistance
or greater suscepti
bility of the embryo to the virus, depend ing upon the interval between inoculation of E. coli (or its endotoxin)
and the virus
(Finkelstein, 1961b). When embryos have been inoculated incubation,
hours
45 has
shown
referred
eggs
have already
results of egg passage on the virulence Newcastle
disease
virus
defective
a high
at
percentage
of the
development
Propagation Ducks
One of the first reports on the modifica tion of Newcastle
disease virus by passage in a bird other than a fowl was made by
Komarov
and
succeeded
in carrying
Goldsmit
14 serial passages by intracerebral
serial
passages
the
embryonic
incubation
have
shown
no
gross
defect.
The virus in chorio-allantoic fluid has subjected to purification
procedures
and chemical analysis (Cunha et al., 1947). These studies have indicated that the haemagglutination to the virus
is proportional Further purification
activity
content.
has resulted in a virus preparation
without
have
who
(1946b)
the virus
(Blattner and Williamson, 1951), and the
and
neural tube and visceral arches (William son et al., 1953). Eggs inoculated at 48
been
of re
in Avian Hosts
of 4- to 8-week-old
hours
been
viewed under the heading "Live Vaccines"
lens
sac
have
on page 97.
lation
auditory
been
to on pages 69 and 70, and the
inoculation of Newcastle
subsequent
disease virus, the results have been either increased
in embryonating
been
ducks.
through inocu
Thirty
reported
by
(1957), and 32 passages by Mitev and Gagov (1958). The virus has also been passaged by intramuscular or sub cutaneous inoculation of ducklings 1 to 10 days old (Seetharaman, 1951b). In other studies, a virulent strain of virus has Syurin
been
attenuated
by
genicity
serial
intracerebral
loss of anti (Martini and Kurjana, 1950).
passage in ducklings
without
Pigeons
property (Edlinger Vaux-Saint-Cyr, 1955). From the genetic point of view, both the sire and the
The susceptibility of pigeons to New castle disease virus was early established by Doyle (1927) and this species has since
dam have apparently
been
any haemagglutinating and
contributed
in rate of viral growth
variation
(Retae/a/., 1963). In embryonating chicken
to the in their
eggs
ference
between
disease
viruses
(Sinkovics, been shown
with
mumps has
been
eggs,
and
inter
Newcastle
demonstrated
has 1957a). An interferon to be present in eggs infected
(Levine, disease virus This interferon has reduced the of Western equine encephalo
Newcastle
1962b). amount
myelitis virus produced Newcastle
used
identifying
the
disease
1943) and for determining the pathogenicity of different strains (Olah and Palatka, 1963). It has also been found possible to carry virulent strains of virus through a number of passages in pigeons by means of intracerebral inocu lation of brain suspension (Olah and 1963). Following nearly 400 pas of the virus by the subcutaneous route in pigeons, some attenuation of
Palatka, sages
virulence
by cells.
disease virus has been main
for
(Beaudette,
to fowls has resulted (Seethara
man, 1951b).
tained by serial passage in duck embryos with
a
attenuation resulting (Seetharaman, 1951b).
Macroscopic
and
for fowls
microscopic
lesions
Doves After 15 intracerebral passages in a species of dove, the virus became avirulent 117
for fowls but retained its immunogenicity
chicks
when
antigenicity
inoculated
and Gagov, in striped
1958). Also, serial passage ground doves has resulted in
considerable strain
(Mitev
intramuscularly
of
attenuation
of virus
Kurjana,
and
(Martini
virulent
a
1950).
Hosts
in Mammalian
Propagation
With the exception of man, Newcastle disease is not spontaneously to
natural
under
mammals
(Verge,
transmissible conditions
1948).
Hamsters Reagan a virulent
et al (1948a, 1949) passaged strain of Newcastle disease virus
300 times in the brains of Syrian hamsters. of the 92nd passage was non
Virus
for chickens, although paraly
pathogenic
sis and death followed the intracerebral inoculation of hamsters. Hamsters inject
with virulent
ed
virus and
intraperitoneal
by the intranasal, routes
subcutaneous
have also been shown to develop signs of or pneumonia or both encephalomyelitis
(Prudovsky et al, 1961). The results of
these
studies
central
suggest
of
abundance
that
susceptible
nervous
system,
there cells
but
is
in
that
capacity to produce virus is relatively
an the
their low.
Bats Reagan
et al. (1952a)
found
the large
brown bat (Eptesicus to Newcastle by
fuscus) susceptible disease virus administered
routes. The virus was re from a number of organs taken death or from bats surviving 14 days. different
covered at
Mice
without
has been reduced
(Tokuda,
the initial inoculation
loss
of
1956). Following of mouse brain, the
virus begins to multiply and becomes non infectious; later the infectivity titre of the tissues rises, but falls again later as the process is repeated (Cairns, 1951). An extensive pneumonia and pulmon ary consolidation the
mouse
by
has been produced Newcastle
disease
in
virus
(Hanson et al, 1951a; Ginsberg, 1951). This effect has been considered to be due to the toxicity of the virus rather than multiplication of the virus in the mouse lung (Davenport, 1952). The toxic effects of Newcastle disease virus in the mouse have been modified by viral interference, by a receptor destroying enzyme, or by xerosin (Grope and Dougherty, 1956). Following the intravenous injection of mice, the virus has been cleared from the blood
by the reticulo-endothelial the liver (Brunner et al, 1960).
cells of
Guinea-Pigs In some studies, it has been possible to make only a few passages in the brains of (Tokuda,
guinea-pigs
1956);
other studies have indicated
whereas
that the virus
could be adapted to this species (Syurin, 1957; Verge and Placidi, 1956), produc ing a non-purulent
disseminated
enceph
(Syurin and Skalinski, 1957). Fowls inoculated with guinea-pig-adapted alomyelitis virus
have developed
have
resisted
HI
inoculation
antibodies with
virus for a period of six months
and
virulent
(Syurin,
1957).
Rabbits Young rabbits
have
ceptible to intracerebral strains of Newcastle
al,
been found sus inoculation of six
disease virus (Reagan
Strains of virus have been successfully passaged in the brains of mice, especially
1954b). Symptoms of irritability and general paralysis resulted. In contrast,
when a susceptible strain of mice (Reagan et al., 1952c) or suckling mice (Tokuda,
the inoculation
1956) has been used. After 43 passages in mice, infectivity for 3- to 5-week-old 118
et
of virus into the anterior chamber of the eye has caused no symp tom in rabbits, although HI antibodies have
been
demonstrated
in
the
blood
TABLE
19 — Tissues used
Newcastle
for In Vitro Propagation Disease Virus (Bankowski, 1964)
of
Tissues
Host
Chicken embryo
Whole undifferentiated Heart and lung
Kidney Liver Heart
Lungs Fibroblasts Skeletal
muscle
Intestine
Chorio allantoic membrane Amniotic membrane Leukocytes (PE) (P) Macrophage (PE) (P) Kidney (PE) (P) Lymph node (F) (L) Kidney (F) (P) Skin and muscle (L) Kidney (PE) (P) Corneal endothelium (PE) (P) WRC-256-Carcinoma (L) L-Fibroblasts (PE) (L) Ehrlich's ascites tumor (PE) (L) Kidney Kidney Lung (F)(P)
Chicken, postembryonic Porcine Bovine Rabbit Rat
Mouse Guinea pig Monkey Human
Heart (L) Uterus (U12)
Maben-epithelial-adeno-carcinoma of lung (PE) (L) H.ep.-2 (epidermoid carcinoma of larynx) (PE) (L) K.B. (carcinoma of nasopharynx) (PE) (L) Hela (carcinoma of cervix) (PE) (L) Detroit 98 (adult sternal marrow) (PE) (L)
Source and type, if known or applicable : L — line cell F —fetal
PE — postembryonic P
— primary
serum (Bonaduce, tration of antibiotics production (Bonaduce,
Following
of
HI
1954c). The adminis has not affected the
Monkeys
antibodies
virus
1954b). alternate
embryos and newborn
in
rabbits
Following passage of Newcastle disease through five different species of
mammals, passage
in
chick
rabbits, Newcastle
disease virus
has been adapted
intracerebral
passage
in
rabbits
to serial
(Mitev
and Gagov, 1960a). After 21 passages in rabbits, the virus lost its pathogenicity for chickens but retained some antigenicity.
the virus has been adapted to
serial passage in monkeys with an encepha litis usually resulting (Collier et al., 1950).
Propagation
in Tissue Cultures
Goret and Provost (1963) have tabu lated the origin of some of the main cell 119
lines used in tissue culture work; and the properties of Newcastle disease virus when
in this
Propagated
of
strain
manner,
retained
virus
virulent
a
its
antigenic
in a number of different cell by Durand
properties. Similarly, the pathogenicity has not been markedly affected (Day et al,
(1962) and reviewed by (1960), Sanders et al. Bankowski and (1964). Bankow(1953) ski (1964) has listed the cell types found susceptible to various strains of Newcastle
field, 1956). A change in the neuropathogenicity of a strain of Newcastle disease virus has been produced by a method
propagated
have been described
cultures
and Eisenstark
Fontanelli
et al.
disease virus (Table 19) and they are also summarized in Anon. (1963b).
Avian Tissue Cells Using plasma
minced and
chicken
embryos
in
Topacio successive passages of
Tyrode's solution,
(1934) reported
31
a strain of Newcastle disease virus. Details
of the cultivation
of Newcastle
disease
1959; Seadale and Winter-
1953; Rusev,
involving the propagation in suspensions of microglial cells obtained from the brains of chick embryos (Piraino and Hanson, 1959). However, in monolayers of chick embryo lung epithelium, there has been little evidence of significant amounts of infectious virus within the cell (Rubin etal, 1957). Using roller-tube cultures of epithelial from chick and cells mesenchymal
virus in tissue cell cultures have since been
embryos,
Pereira
described by Seiffert (1955) and Buthala (1957), and reviewed by and Mathews
observed
a progressive
Csikvary et al. (1961). In addition, the effect of the virus on the metabolic path ways of embryonic chicken fibroblasts has
reached
been
examined
by
and
Magee
Sagik
(1959). No difference has been detected in the infectivity peak densities of Newcastle
virus
content
and
(1954) in the of the fluid phase which
a maximum
Gompels
increase
between the fourth
and eighth day. Some cultures have shown changes in the morphology of infected epithelial cells (Bang, 1953a; Hotz and Bang, 1957). In the chorio characteristic
membrane,
allantoic
virulent
disease virus when propagated in embryo of chicken, duck fibroblast monolayers
troyed the host cell cytoplasm.
and goose (Stenback and Durand, 1963). Zuschek et al. (1958a) reported that a
membrane
maximum in
isolated
suspended
titre of virus, when propagated
chorio-allantoic in Tyrode's
membranes
solution,
was ob
tained at pH 9.5 in the presence of 0.2 gm. CaCl,
and 0.4 gm. KC1 per litre. Later Zuschek et al. (1958b) described the effects of various concentrations of antimetabolites on the growth of New castle disease virus. They also found that
the
virus
membranes
best
in
chorio-allantoic
incubated
at
42°C, although
grew
haemagglutinin
was
not
demonstrable
(Zuschek et al, 1959). In trypsinized chick embryo tissue cul ture, Newcastle disease virus has given a maximum
infective
ml. in 24 hours 120
titre of 10-7-5 per 0.1
(Csikvary
et
al,
1961).
strains
of
disease virus have rapidly des
Newcastle
Less viru
of the with virus sometimes being released from cells without causing cellu lent strains have caused hyperplasia
lar destruction (Bang, 1953b). Certain cultures of chick embryo cells have yield ed about
300 particles
(Kohn and Goldwasser, microscopy ease
virus
minutes embryo
of virus per cell 1957). Electron
has revealed particles
after
dis
Newcastle
in the cytoplasm
30
inoculation of chick (Mussgay and Weibel,
the
cultures
Thus it has been concluded that morphologically intact Newcastle disease virus particles are capable of entering a 1962).
cell.
Virus propagated
in
chicken
embryo
skin tissue cultures has shown greater viru lence to young chicks when compared the same strain
of virus
with
propagated
whole embryo tissue culture medium
in
(Day
NEWCASTLE
DISEASE VIRUS IN AVIAN TISSUE CULTURE
HA Titre
LD50
Hour (Redrawn from Csikvary et al, 1961)
Figure 19.
et al.,
1953). In chick embryo tissue cul
tures,
the
and immunizing activities have developed
in embryonating and eggs; following a number of 6-hour multiplication cycles, the maximum infec
in a parallel manner
tive titre occurs
though
infectivity,
haemagglutinating (Cessi,
1959);
virus titres have been lower
those obtained
by propagation
al than
in chick
determined afterwards,
at 20 hours
(Csikvary trated in Figure lation
19.
1961),
(Strizhachenko, 1961). Studies using suspensions of chicken embryo tissue cells have also shown that the inoculation
to the 50th hour post inoculation. The HA titre of the culture has reached a maxi
of a virulent strain of Newcastle
mum after 38 hours.
embryos
disease
virus results first in a drop in virus titre as
titre has remained
post inocu
as illus This peak infective
al.,
et
A
comparison
largely unchanged
of the propagation
up
rates 121
of a virulent strain of Newcastle disease virus and Strain F virus (Asplin, 1952)
infective
has shown that the latter virus is liberated
divide (Wheelock and Tamm, 1961). The virus has been found to spread from cell
more slowly from infected chicken embryo cells
the virulent
than
virus
(Mussgay,
1960). Another difference noted, when cultures of cells from chicken trachea have been used, is that the cells contained virus for 12 weeks and a lento-
a virulent
for at least 16 weeks after (Morehouse et al., 1963a). Ultraviolet irradiation of cells has re sulted in a higher yield of virus per cell genic
virus
infection
and
(Rosenberg
Rosenbergova,
1962).
from explants of trachea, lungs and kidneys of chickens have been maintained for 4 to 16 months without Cell cultures
(Morehouse et al., 1963a). Adult fowl spleen tissue with added fowl subculture
serum has been used
in the
successfully
of Newcastle
propagation
(Hayashi, 1956; Hayashi
disease
virus
et al., 1962).
Mammalian Tissue Cells Some strains of Newcastle distinct
produce
lines without
disease virus
in human
plaques
any previous
cell
adaptation
of
the virus to the plaque technique. The use
of
a
of agar in the over staining of the after removal of the agar, has
low concentration
lay followed by a non-vital cultures
the identification
permitted
and titration
in HeLa cells of strains and mutants of Newcastle
disease virus
A
strain
variant
(Zavada,
produced
1963). by continued
passage in HeLa cells has been found non pathogenic retained
to
fowls,
its immunizing
it
although properties
has
(Hal-
lauer,
1958). Using the
technique,
fluorescent-cell
Newcastle
maximum
counting
disease virus
pro
titres in
6
to 7 hours
(Wheelock, 1963), or in 24 hours (Tyr rell, 1955). In HeLa cell monolayers, the addition of Newcastle disease virus has increased the rate of release of potassium (Klemperer, 1960); but the production of 122
virus
the
to cell
has not per se interfered
of an infected
ability
the
through
cell
to
intracellular milieu
(Wheelock and Tamm, 1961), and pro gressive inhibition of mitotic activity in infected
cells
continued
has been
correlated
of viral
production
with
antigen.
Cytopathic changes have developed after mitotic activity has decreased to low levels (Wheelock and Tamm, 1961). In individual cells of HeLa cultures, non-infectious
have been
haemagglutinins
Morgan, 1961), and also the synthesis of Newcastle dis ease virus ribonucleic acid (Wheelock, Bankowski 1963). (1964) showed that the propagation of the Bl strain in HeLa cell cultures resulted in a predominance of infection of individual cells (Figure 20). In cell cultures of guinea-pig bone marrow, formation of new viral antigen has begun four hours after infection (Jerushalmy et al., 1963). The virus has been propagated in cul tures of embryonic human lung (Chapronieve and Pereira, 1955), human car cinoma cells (Pigoury et al., 1962) and human leucaemic bone marrow cells, (Henle, 1960). Using fluorescein-conjugated antibody stain, Johnson and Scott (1964) demonstrated the presence of Newcastle disease viral antigen in the nucleus and cytoplasm six hours after infection of HE p-2 cell coverslip cultures. Studies with Newcastle disease virus in identified
mouse
L
(Bader and
strain cells have shown by an
adsorption
duction has been shown to begin in HeLa cells 4 to 5 hours after infection, and to reach
with
L
that the
cell of a single
in
fectious particle was sufficient to result in loss of cellular the cytotoxic
integrity.
In
these cultures
effect was accompanied
synthesis of large amounts tious haemagglutinin
of infectious other occurred
virus
and small
(Wilcox,
by
of non-infec amounts
1959a). In
cultures,
has cytopathogenicity only when the concentration of
virus exceeded one egg infectious
dose per cell (Mason and Kaufman, 1960).
srnBij
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Infection of L cells with small amounts of Newcastle disease virus has resulted in
crease in this cytopathic
cell
produced
resistant
populations
to
the
virus.
found to contain small amounts of virus which persisted at extremely low levels for extended periods (Paucker et al, 1962; Wilcox, 1959b). A scheme of the events occurring upon initial infection of L cells by New These
cultures
were
castle disease virus
indicates
that a cell
which adsorbs an infectious virus particle will produce infectious particles (Rod riguez and Henle,
disease virus
gated well
(Kono,
tissue cultures, has not
propa
in others the
1962);
virus has persisted for 1 1 days (Klapotke, 1952).
In calf kidney tissue, Newcastle disease virus
has been either incompletely
cyto-
(Dannacher and Fedida,
1962) or has shown a decrease in this effect (Rus-
pathic
seff, 1962). The virus has shown
in cultures of rabbit endothelial cells (Oh, 1961). mammalian
for the propagation
corneal
tissue cultures
of Newcastle
mul
used
disease
virus have included swine tissue (Kumagai et
al,
1958; Bengelsdorff
and Schneider,
1963b), monkey cells (Andre and Audebaud, 1960), Ehrlich ascites cells (Adams et al, 1955) cells (Bang and War
and Prince, 1957; Flanagan, and rat carcinoma
wick, 1957). The virus has also been pro pagated
in seven other
cultures
(Stenback
Cell cultures bryo lymph
mammalian
cell
and Durand, 1963).
derived
from swine em
nodes have been found to be
sensitive to small amounts of virus and to support the development
by the addition
(Matumoto of fresh
cultures
al,
et
swine
an in
effect has been
of hog cholera
In
1961).
Newcastle disease virus has produced pathic
cyto
which first appeared as granules and vacuoles (Shi-
changes
cytoplasmic mizu,
tissue
kidney cortex,
etal,
1957).
The danger of introducing Newcastle disease virus by employing chicken plasma in any tissue culture
system was
clearly
by Chanock (1955) during studies on variants of poliomyelitis virus.
This latter virus was serially propagated in monkey kidney epithelial cells with the production of granular degeneration. Cells which escaped the cytopathogenic effect of the Newcastle disease virus were found resistant to the cytopathogenic effect of some types of poliomyelitis virus. Tortoise Tissue Cells
appreciable
tiplication Other
virus
al, 1961);
et
demonstrated
1964). In some mouse embryo
Newcastle
(Kumagai
vation
of high titres of
disease virus has undergone passages in monolayers of
Newcastle
ten consecutive kidney
epithelium
cells
of the
tortoise
(Testudo graeca) (Schindarow and Todorov, 1962). There was a pronounced cyto pathic effect and nuclear
inclusion bodies
appeared in the infected cells. The forma tion of these inclusion
bodies was neutral
ized by immune serum from a fowl (Fauconnier, 1963).
Propagation Sarajew
in Yeast Cells
(1954)
disease virus
cultivated
Newcastle
in yeast cells in liquid
and
solid culture media. Maximum multiplica tion of the virus occurred after 24 hours at 16° to 28°C.
Repeated subculturing of
virus (Morehouse et al, 1963b). In swine testicular cell monolayer cultures, New
Newcastle
castle
for fowls with no detectable loss of anti
disease virus
has produced
cyto-
pathic changes after six to eight days culti
124
disease virus for 25 generations
in yeast cells resulted in loss of virulence genicity against virulent
virus.
PREPARATION Preparation of Vaccines from Embryonating Eggs Diseases
Egg-Transmitted
Factors that lead to the production of chicken
normal
embryos
have been the
subject of an extensive review by Landauer (1961) and are summarized in Anon.
Not
(1959,
only the
general
1963b). health of the embryo, but also the import ance of infectious agents in the egg has
Thus,
studied.
been
(1956)
found
a
Burmester
commercial
et
al.
New
live
castle disease vaccine caused a highly signi
of visceral lymphomatosis chickens; from field evi in inoculated dence, Gallego Piedrafita (1952) sug formolized gested that a contaminated Newcastle disease vaccine was the likely cause of the appearance of fowl paralysis in a number of vaccinated flocks; Hejl and Faber (1959) found bacterial contamin ants in 35 of 53 samples of commercially
ficant incidence
available Newcastle
disease vaccines;
Wil
of organisms (1957) identified faecal origin and various fungi in other vaccines of egg embryo origin; Thompson (1954) thought that the causal agent of disease was trans chronic respiratory mitted in a Newcastle disease vaccine, and demonstrated that a culture of Newcastle disease virus contaminated with the agent of chronic respiratory disease could be liams
carried
without
through the
several embryo
contamination
passages
being
sus
Kruger
(1961) emphasized the possible transmission of leucosis virus. Cottral (1952) has summarized diseases of birds transmitted through eggs (Table 20), and he has emphasized that, whereas pected;
the different etiological
agents in eggs have
been found, it has not been established in all cases that the agents can cause dis ease
in hatched chicks
transmission.
as a result
of
egg
In discussing the report of
OF VACCINES Cottral (1952),
Cox
has com
(1952)
production of vaccines, involving millions of eggs, no virus agent has been contaminating detected by the various types of safety mented that in the commercial
agents must
sible
potentially
egg trasmis-
Nevertheless,
employed.
tests
be regarded
as
being
vaccines.
able to contaminate
Techniques used to inoculate embryo nating eggs have been discussed on pages 68 and 69. Both the Roakin and Bl
of vaccine virus have produced gross defects in chick embryos when eggs have been inoculated at less than 72 hours incubation (Williamson et al., 1953). strains
When inoculated the
Bl
strain
at ten days
curling of embryos
incubation,
stunting
has produced
and
on the fourth (Hitchner et al.,
dying
succeeding days 1951a). This abnormal development may be due to changes in the flow of fluids from the embryo into the allantoic cavity. When eggs inoculated with a virulent and
strain
of Newcastle
disease
at 32.2°C,
virus
have
the embryonic
been incubated survival time has increased from approxi mately 40 hours to 70 hours (McLimans et al., 1950). When large doses of virulent Newcastle disease virus have been inocu eggs, a number lated into embryonating
have survived an otherwise lethal infection (Karrar, 1963). In these resistant embryos, there has been little or no multiplication of virus. The exact cause
of embryos
of this abnormal ascertained,
behaviour
however
has
not been
viral toxicity has
been considered likely.
The Influence of Parental Immunity on the Preparation of Vaccines Antibody passively transmitted from an dam has an appreciable in on the embryonic mortality rate following the inoculation of Newcastle immunized
fluence
125
20 — Diseases
TABLE
of Birds Transmitted Through Eggs (Modified from Cottral, 1952) Etiological
Disease
Principal hosts
agent
Avian lymphomatosis Avian encephalomyelitis Newcastle disease Infectious sinusitis Psittacosis Pullorum disease
Chickens
Virus-like Virus
Chickens
Burmester et al.. 1956 Van Roekel, 1959
Virus Virus
Chickens Turkeys
Jerstad et al.,
Virus Salmonella pullorum Mycobacterium avium Salmonella gallinarum Salmonella
Parakeets
agent
Lancaster,
1963a
1949
Avian tuberculosis Fowl typhoid Paratyphoid infections Chronic respiratory
Chickens Chickens, ducks Ducks, pigeons. chickens, turkeys Chickens, turkeys
Thus, with the Bl strain, Hitchner et al., (1951) found that em disease-immune bryos from Newcastle of 33 per parents showed a mortality inoculation;
whereas susceptible embryos gave a mort
ality of 67 per cent on the fourth day post inoculation. The corresponding mortality pattern for Strain F virus was 15 to 17 per cent for immune embryos six days post inoculation, and 75 per cent for sus ceptible embryos four days post inocula tion (Lancaster, data; Rao unpublished andAgarwal, 1960). of New Reports on the preparation have indicated
Wilson, 1947 Fahey and Crawley,
1954
disease virus.
castle disease vaccine
Beaudette, 1 925
turkeys,
Mycoplasma gallisepticum
day post
Meyer, 1934 Rettger and Stonebunn, 1 909 Feldman, 1959
Chickens
sp.
disease
cent on the sixth
Selected references
that
no significant effect upon the final virus titre. However, for the propagation of Bl vaccine virus, eggs from immunized hens have been considered unsatisfactory
(Szakmary and Toth, 1963). Similarly, Topolnick and Beganovic ( 195 1 ) have reported higher
haemagglutination
titres
in
by immune hens.
Using the Bl strain of virus, Reta et al. found that both the sires and dams
( 1 963 )
made statistically to the virus
significant
contributions
titres in the amnio-allantoic
fluid, as determined was interpreted
by the
HA
test. This
to indicate that variations
in the rate of viral growth in eggs was in
from vaccinated hens could be used (Daubney. 1953; Brandly et al., 1946c).
fluenced by genetic effects. With Strain F virus, no difference
Furthermore,
been detected in the
eggs
no
difference
has
been
eggs
hens than in eggs laid
laid by non-immune
HA
titres
of
has
fluids
found in the viral multiplication in 1 1 -dayold embryos from non-immune and im mune hens (Liu and Bang, 1953).
from susceptible embryonating eggs com pared with eggs from immune dams (Lan
Hitchner et al. (1951) reported a slight lag of approxi
whenever
mately two hours in immune embryos, al
in the preparation
though the passive antibodies
1957a).
With the Bl
126
strain,
present had
caster,
data). Nevertheless,
unpublished possible,
embryos
free
from
parental antibody should probably be used
of vaccines (Lancaster,
AMOUNT OF NDV (Bl STRAIN) IN ALLANTOIC FLUID AND EMBRYO TISSUES FOLLOWING ALLANTOIC SAC INOCULATION OF NON-IMMUNE EMBRYOS Log Dilution - 9
-7
0/0
8
16
24
32
48
60
84
72
96
Hours after Inoculation (Redrawn from Liu and Bang, 1953)
Figure 21.
Storage of Virus Material The ability of strains of Newcastle dis virus to survive for long periods faci litates the repeated production of vaccine from the same initial virus suspension. The virus in amnio-allantoic fluid has re mained viable for over 28 months at 1°C ease
(Bonaduce, virus
has
1950c). However, been inactivated
lyophilized
within
10
months when stored at 37°C (Hofstad and
Yoder, 1963).
Use of Extra-Embryonic Fluids with or without Suspensions of Embryonic Tissues Vaccines consisting of chorio-allantoic fluid mixed with some embryonic tissue have been prepared Szakmary
and
Toth,
(Rao
et
1963).
al,
1963;
However,
Gagliardi (1960) has reported that the virus-containing fluid loses some of its infectivity and immunogenicity when mixed with yolks from immunized hens. 127
AMOUNT OF NDV (CG179) IN ALLANTOIC FLUID, BLOOD AND EMBRYO TISSUES FOLLOWING ALLANTOIC SAC INOCULATION OF NON-IMMUNE EMBRYOS Log Dilution
-10-
I
I
I
I
I
I
I
0
8
16
24
32
40
4*
Hours after Inoculation
(Redrawn from Liu and Bang, 1953)
igure 22.
This effect may be due to specific antibody
(Figure 21). Thus, the lungs of 16- to 17embryos have been found very
a the yolk and yolk should not, therefore,
day-old
ie included
susceptible to the
in the vaccine.
With the Bl strain of vaccine virus, the imount of virus in the tissues of the emsac inoculation >ryo following allantoic
1953). maximum
emains about 2.5 to 3.0 logs lower than
particles
he amount 28
of virus in the allantoic fluid
Under
Bl
virus
certain
(Liu
yield of Newcastle
per allantoic
and Bang,
conditions,
the
disease virus
cell has been about
(Horsfall, 1956). Atanasiu and Gareau (1952)
1,000 found
the infectivity of Newcastle
that
disease
shortly after the embryos. Thereafter,
virus reached a maximum death
of inoculated titre fell rapidly if the eggs were at 37°C. Contrary results have been
HA
the kept
al.
who
by Chang (1956) that both the infectivity and hae-
reported found
et
magglutination
titres of the allantoic fluids
continued to rise following further incuba tion up to 24 hours after death of the em bryos. A decrease in both titres began 36
al,
death (Chang et
hours after embryonic
in the field for periods
transported
In
three
weeks.
where
adequate
is not available,
countries
up to
or districts equipment
freeze-drying
fresh vaccine might, per
and, with care, used in the field (Lancaster, 1956). Nevertheless, in India, arranging for field workers to have a continuous supply of the fresh (undried) vaccine and ice for haps, be prepared
satisfactorily
its preservation,
was the most difficult part
of a Newcastle disease vaccination paign (Naidu, 1959).
cam
1960).
The immunizing capacity of an inacti prepared from egg fluids corresponds to the HA titre of the fluid. vated vaccine
HA
Thus, the
has been considered
test
a
control procedure in the prepara tion of vaccines from embryonating eggs (Cordier et al, 1950; Hanson et al, 1947; Hill, 1957). Nevertheless, the embryo
Preparation of Inactivated Vaccine
useful
LD50 titration an is
is much
more sensitive as
indicator of the presence of virus than the HA test (Hanson et al, 1947). In the case of a virulent strain of New
The action
acid
of virus in the embryonic
tissues has close
virus
fluid in
(Liu
the amount in the allantoic
and Bang, 1953),
as illustrated
Figure 22. However, Cordier
al.
et
(1950) have reported that the virus from
HA
pulped embryos has reached a higher titre
than that from the amnio-allantoic
fluid. The influence of the route of inocu lation of Newcastle
disease virus on selec
tive infection of the fully susceptible em bryonating egg reported by Hanson et al.
(1947)
is shown
in Figure 15 (page 71).
number
of physical
has been reviewed
clude an irreversible
179), the amount
ly approached
a
by
Pollard (1960). It has been considered that true inactivation of a virus particle, that is, complete destruction of infectivity and capacity of multiplication, must in
(CG
castle disease virus
of
agents on viruses
(Gard, 1960). inactivation
cannot
be reversed
longed storage
change in its nucleic
Thus, the degree of by
beta-propiolactone
by dilution or pro
(Lo Grippo,
Some of the problems
1960). associated with
of virulent Newcastle dis virus have been mentioned on page 00. One of these problems relates to the presence of active virus residue in an other wise inactivated product. In this connec tion, a report by Cordier et al. (1950) has indicated that the adsorption of live the inactivation ease
virus
on aluminium
hydroxide
results in
some loss of virulence of the non-adsorbed
Preparation of Fresh and Lyophilized Vaccine In particular
situations,
it has been im
of lyophilized vaccine of either lentogenic or possible to prepare
mesogenic 1957a).
adequate amounts
vaccine
strains
(Lancaster,
Under tropical conditions,
(undried)
Strain
F
vaccine
has
virus. Use of infected amniotic-allantoic inactivated with formalin and diluted
fluid 1 : 80
in a thin aluminium hydroxide
suspension has resulted in better immunity than the
of thicker commercial vaccines (Perez1962). The use of infected mem branes and chicken embryos apparently
use
Rebelo,
fresh
increases
been
cine.
non-active
proteins
in the vac
129
Preparation of Vaccines from Virus Propagated in Tissue Culture
incubated a
and tubes rotated
stationary
cultures, infectivity
and
the haemagglutination titres
were
lower and
strains
been
have
cultures
in
propagated
Schneider,
and
(Bengelsdorff
tissue
1963b; Brandt,
1961; Cessi, 1959; Gelenc-
zei and Bordt,
1960; Mayr, 1961; More
house
1963a;
al.,
et
Rossi,
1961a,
b;
1960). This by Fontanelli,
and Pomerov,
Subramanyam
subject has been reviewed
etal. (1960). Russeff (1962) considered that the use of calf tissue culture eliminated the risk of contamination by avian leucosis virus in the vaccine. However, a number of in have reported
vestigators
ment of haemagglutinin
poor
of the lentogenic
antigenicity
develop
and some loss of and other
strains of viruses when propagated sue culture.
in tis
Thus, the virus titres obtained in chicken
by propagation
embryo
fibro
corresponding
titres attained
the took
than the
longer to reach their maximum
To overcome some of the disadvantages inherent in the use of embryonating eggs (Bankowski, 1958), Bl and F vaccine
in
roller drum. In the case of the stationary
in the cul
tures on the shaking machine.
The virulent field strain, designated California 11914, has been propagated in suspensions of chick embryo cells (Bank 1957; Bankowski and Boynton, owski, 1948). Serial passage resulted in attenua tion without loss of antigenicity (Bank 1958) and this adapted strain has been designated "TCND". The virus could owski,
in cultures of HeLa or pig kidney cells (Bankowski, 1958; Bankowski and Hyde,
also be propagated
cells and in bovine 1957;
Provost
were formed
et al., Syncytia 1962). in HeLa cells, as shown in
Figure 20 (Bankowski, 1964). One fea ture of this tissue culture virus was its in ability to spread to susceptible in-contact (Bankowski et al., 1958a). One
chickens
dose of the vaccine muscularly
administered
intra
to susceptible chicks at 5 days
blasts have been lower than those obtained
of
by using chicken embryos (Strizhachenko,
at least 13 weeks
Peeter-
immunity lasting (Bankowski et al., 1950, at 39 days of age 1958a). Revaccination following Bl vaccine at 5 days of age resulted in an immunity against virulent
or Hertfordshire strain 1963) (Markovits and Toth, 1962). Good im
virus lasting 93 weeks (Bankowski et al., 1963). Two doses of the vaccine gave
1961),
although
no loss of antigenic pro
perties has been noted for the mesogenic
Komarov strain
and
(Huygelen
mans,
munizing has
activity
of tissue culture virus by Hallauer and
also been reported
Kronauer (1960) and by Rossi (1961b). Iwasaki (1954) examined a number of factors that influence the propagation of Newcastle disease virus in chick embryo tissue cultures and compared ods of virus
propagation
three meth
using:
culture
tubes attached to a shaking machine, tubes
age
or over induced
immunity
lasting from 33 weeks
owski,
1957) to second vaccination
weeks
(Bank
after
1960). strain and a crystal violet vaccine,
tissue
the
(Bankowski and CorWhen compared with the
stvet,
Bl
101
culture
vaccine
gave
the
appreciably
protection against virulent virus (Bankowski and Hill, 1954; Bankowski etal, (1958b). greater
STORAGE AND TRANSPORTATION OF VACCINES Studies have been made on the storage
of fresh
(non-lyophilized) virus, either with the addition of preservative (Bella, 130
1960) or without. Strain F virus, as fresh unpreserved allantoic fluid, has been kept at a temperature
of
4° to
6°C for at
least
21 — Survival of Newcastle Disease Strain F Virus at Different Temperatures (Modified from Monda and DeBonis, 1959)
TABLE
Vaccine at 1 5" to 27'C
Vaccine at 37°C
Length of exposure (days)
HA
% chickens
titre
surviving1
25
80 80
100 100
25
35
45
80 40
80 25
45
55
65
40
10
75
20
0
65 75
Length of exposure (days)
HA
% chickens
titre
surviving1
40 40 40
35 55
100 80 0 0
20 20
—
10
1 % vaccinated chickens in groups of 5 surviving challenge
without loss of immunizing
six months property diluted tained
1957a). Fresh, un Mukteswar virus vaccine has re (Lancaster,
its immunizing property
after
exposure of 7 days to an atmospheric
an
tem
of 24° to 32°C (Generoso and Mendoza, 1950), or for 33 days at a temperature of 18° to 22°C (Gualandi, 195 1 ) . Lyophilized Mukteswar vaccine has remained viable for 30 days at room perature
1957). A formolized vaccine adsorbed onto aluminium hydrox temperature
(Datta,
ide when frozen in a refrigerator
and sub
sequently
thawed, has been found
antigenic,
although
to be
there had been some
of antigenicity (Placidi and Santucci, 1953b). However, a formolized vaccine with an adjuvant, following storage for
loss
600 days at room temperature, capable of stimulating
has
been
protective immunity
in fowls (Walker and McKercher, 1954a). With the Bl virus, a frozen vaccine pre pared with 20 per cent glycerin has main tained an adequate titre during
12 months
storage
(Reising and Hitchner,
Thawed
vaccine preparations
1954). have gradu
ally lost titre when stored at 5°C.
Using
lyophilized
amnio-allantoic
fluid
Strain diluted
F
1
virus
in
in
in
2
milk, Monda and DeBonis found the immunizing power to be (1959)
skimmed
unimpaired
after 35 days storage at 15° to
27°C (Table 21). The resistance of Strain
F
cine to adverse conditions
has been re
virus
vac
ported by Torlone (1956) who found that the virus survived two months at 4°C after being kept initially for 48, hours at
F
fluid has stable at temperatures of 25° and 37°C for 48 hours (Torlone,
25°C. Strain
virus as allantoic
also been relatively
Newcastle 1955). Nevertheless, disease virus is susceptible to exposure to daylight. Thus, unfiltered amniotic fluid infected with the virus, when exposed to daylight
for four hours at 4°C has shown a de crease in titre of about 3 log units (Skinner and Bradish, to light 1954). Exposure therefore could cause serious errors in infectivity titrations. More comprehensive
studies have been
made on live Newcastle
disease virus vac
lyophilized product. The immunizing property of lyophilized Strain F virus has been unimpaired after storage for one year at -22°C or nine months at 0°C (Lancaster, 1957a), as shown in Table 8. Similar material, when stored for two years at 4°C has shown a loss of less than one log unit in virus potency as determined by chick embryo infectivity tests (Anon., 1961b). Other live lyophilized vaccines have been stored for over one year at -10°C without loss cines prepared
as a
13!
VIABILITY OF KOMAROV STRAIN NEWCASTLE DISEASE LYOPHILIZED VACCINE Log Dilutions
-9
-
1
I
I
I
I
I
I
I
i
1
3
5
7
9
11
13
15
Weeks
(Redrawn from Thorite and Mac Laod, 1960)
Figure 23.
of virus concentration
(Borzemska
et
al,
The Bl strain, when diluted
1:10,000
1961). Egg titrations have shown that, with desiccated vaccine of the Komarov
in tap water, has remained viable at room
strain,
buhl
the virus
titre
has not
dropped
for
temperature
al,
et
at least 11 hours
1955),
although
(Lugin-
the
same
greatly during 12 months storage at -15°C
strain at 37°C has shown a lack of stability
(Thome and MacLeod, 1960). At room
(Fernandez Espinosa
temperature,
the same dried vaccine
shown a progressive
has
decline in titre aver
aging about half a log per week over eight weeks. Thus, Thorne and MacLeod
(1960) concluded
that this vaccine could
withstand field temperatures up to a week without
the titre falling below the stand
ard (Figure 23). Kruger (1961) has sug that the storage time should be gested three to nine months, depending on condi tions. 132
et
al,
1961 ).
on the of the virus. Thus, an increased concentration of New castle disease virus in amnio-allantoic Studies
purification
have
been
conducted
and concentration
fluid has been obtained
by adsorption
on
followed by elution with buffered saline (Zinca et al,, 1960) or by thermo-diffusion (Medgyesi, 1958, 1951). Purification of the virus has been aluminium
hydroxide
obtained by calcium graphy
phosphate chromato
(Reda and Rott,
1962)
or by
thermodiffusion
(Medgyesi, results
matographic
have
one strain of Newcastle
1951). Chroindicated that
disease virus con-
tains
at
least
two
Details for the testing of Newcastle vaccines
have
been
dis by
reported
one
of
non-infectious
OF VACCINES
TESTING AND STANDARDIZATION ease
components,
which may be relatively (Wilson, 1962b).
when the ratio of mesogenic to lentogenic virus was 1 : 1 0,000. This is the same dilu
( 1963 ) and are also given in
tion detected by Olah and Palatka ( 1963 ) . In the Federal Republic of Germany,
and 1963b).
the manufacturers
Additional information on the testing of vaccines has been provided by Olah and Palatka (1962, 1963) using the intracere bral inoculation of pigeons. These authors showed that a 1:10,000 dilution of viru
been obliged to test the vaccine according purity,
lent
chicks.
Gehring and Geiss
Newcastle
detected in a
Bl
(1958) and Oshel Anon. ( 1 962c
disease
virus
could
be
vaccine; whereas virulent
virus alone produced symptoms and death
to prescribed
of the Bl vaccine have The
procedures.
tests
for
safety and efficacy have been out
lined by Fritzsche
(1963) and involve the
use of embryonating
eggs
and
day-old
In West Germany, Government testing of Newcastle disease vaccines has been in operation
since
1959
(Eissner,
Thus, it
1961).
appeared that large quantities of
Bl
interfered
with the
The need for careful regulatory control of poultry vaccine manufacture has been emphasized by Brandly (1950), Johnson et al. (1954a) and Levine (1962a). In
in pigeons
at a
dilution of
to a certain
10-6.
extent
virus
infectivity of virulent virus; a finding simi lar to that of Rosenwald et al. (1959). Olah and Palatka (1963) concluded that the pigeon
was
more
reliable
than
the
chick for testing the safety of B 1 However, it must be emphasized that the day-old chick intracerebral patho
addition,
Eissner
standardization
standard,
day-old
philized
vaccine.
that commercial
genicity index has been used to distinguish lentogenic
and mesogenic strains of New
castle disease virus over a wide range of
(Hanson, 1956). In
virus concentrations
(Hanson, 1956) has reported the detection of a mesogenic virus in a mixture addition
(1961)
has discussed
based on an adsorbed lyoand he has suggested vaccines
not less than 40 protecting
should
contain
units per ml.
The practicability of using a single inacti vated standard for the calibration of both inactivated
and live vaccines has been dis
cussed by Stableforth
A
(1961). for examining live virus summarized in Table 22.
procedure
vaccines is
TABLE 22 — Summary of Test Requirements for Live and Modified Live Virus Newcastle Disease Vaccines (Van Houweling, 1963) Salmonella Extraneous bacterial
contamination
Extraneous pathogenic viral contamination Chick safety Virus content Immunizing capability
Must not be present when examined by culture. Must not be more than 10 colonies per bird dose based on plate count. Must be none based on either bird or embryo inoculation. Not more than 2 of 25 chicks, 5 days old or less, may develop severe vaccination reaction. There must be 10 s EIDS0/CC. after incubation of the sample for 7 days at 37°C. 8 of 10 vaccinated birds must be completely protected by one dose ; 8 of 10 controls must be susceptible to the same challenge dose. 133
TABLE
23 — Summation
of Thirty-two Potency Tests on One Batch of
Inactivated Newcastle
Disease Vaccine (Coid et al., 1963)
Dilution of vaccine 1 :25 1 : 50 1 : 100
Controls
Chicks surviving /Chicks challenged Place "A" 123/156 (79%) 87/156 (56%) 60/156 (39%) 10/341 (3%)
Place "G" (72%) (53%) 85/227 (37%) 18/423 (4%)
161 /223 121 /227
The results of 32 potency tests on a batch of BPL-inactivated vaccine during a period of 37 weeks have been reported by Coid et al. (1963) who showed that doubling the amount of antigen increased the percentage of chick
cines. Further study and agreement is re
ens protected
quired,
single
At
(Table 23).
the present time, tests
and criteria
for the safety and potency of Newcastle disease vaccines are not uniform through out the world. However, appreciable
134
quantities of Newcastle disease vaccine are traded commercially between different countries, table
and more uniform and accep
standards
internationally
need
to
be
for Newcastle
established disease vac
so that poultry raisers in regions of the world where vaccination against
Newcastle
disease
is
permitted
can
be
with the most suitable antigens for the immunization of their flocks. provided
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