Veterinary World, EISSN: 2231-0916

RESEARCH ARTICLE
Available at www.veterinaryworld.org/Vol.10/September-2017/10.pdf
Open Access

Pathology and immunohistochemistry study of Newcastle disease field

case in chicken in Indonesia
Etriwati1, Dewi Ratih2, Ekowati Handharyani2 and Surachmi Setiyaningsih3

1. Laboratory of Pathology, Faculty of Veterinary Medicine, Syiah Kuala University, Banda Aceh, Indonesia; 2. Department
of Clinic Reproduction and Pathology, Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor, Indonesia;
3. Department of Animal Disease and Public Health, Faculty of Veterinary Medicine, Bogor Agricultural University,
Bogor, Indonesia.
Corresponding author: Etriwati, e-mail: etriwati.2102@unsyiah.ac.id
Co-authors: DR: dewira@apps.ipb.ac.id, EH: ekowatieko@apps.ipb.ac.id, SS: surachmi@apps.ipb.ac.id
Received: 21-04-2017, Accepted: 18-08-2017, Published online: 13-09-2017

doi: 10.14202/vetworld.2017.1066-1071 How to cite this article: Etriwati, Ratih D, Handharyani E, Setiyaningsih S
(2017) Pathology and immunohistochemistry study of Newcastle disease field case in chicken in Indonesia, Veterinary
World, 10(9): 1066-1071.

Abstract
Aim: The aim of the study was to examine pathology and the distribution pattern of Newcastle disease virus (NDV) in
internal organs of chickens from a field case using immunohistochemical staining.
Materials and Methods: 10 groups of broiler, layer, and domestic chicken were collected from necropsy room Division
of Pathology, Bogor Agricultural University. These chickens were originated from West Java and collected based on
pathologist diagnosis as suspect of Newcastle disease (ND). They were subsequently confirmed positive of ND with real-
time-reverse transcription polymerase chain reaction assay. The respiratory, circulatory, digestive, lymphoreticular and
central nervous systems were collected for histopathology examination.
Results: The gross pathology and histopathology changes were tracheitis, pneumonia, pericarditis, myocarditis, catarrhal
proventriculitis, catarrhal enteritis, typhlitis, perihepatitis, pancreatitis, nephritis interstitial, splenitis, atrophy of Bursa
Fabricius, and encephalitis.
Conclusion: The distribution pattern of NDV in internal organs of chickens from a field case in this study is similar with
a previous reported pattern in systemic cases of the internal chicken organs. High intensity of immunohistochemistry stain
result was detected in trachea, lung, proventriculus, duodenum, cecal tonsil, kidney, and brain.
Keywords: broiler, domestic chicken, immunohistochemistry, layer, Newcastle disease.
Introduction
The emergence of new genotype variant is esti-
Newcastle disease (ND) is an important avian mated from global epizootic and genomic sequence
dis- ease in Indonesia. ND has spread all over changes in low- and high-virulence NDV indirectly.
Indonesia and cause massive loss in poultry business. This hypothesis is that NDV genotype change hap-
The disease is spreading rapidly, both morbidity and pened at the same time in various places world-
mortality of ND virus (NDV) infection can reach wide [3]. ND outbreak in Indonesia happened first
100% which known to have infected over 200 bird time in Java, island on 1926 [2]. Ever since the out-
species [1]. The caus- ative agent of the disease is break, vaccination program management has been
NDV also designated as avian paramyxovirus Type-1, implemented, however until now Indonesia is still
genus avulavirus. NDV is a part of ribonucleic acid an endemic area of ND. Moreover, virus circulation
virus with the single stranded genome and negative can be detected throughout the year. ND outbreak in
polarity. Based on its virulence, NDV is divided into 2009-2010 in commercial chicken caused 70-80%
four groups, which are velogenic viscerotropic (Asian mortality [4]. Pathological findings, phylogenetic
type) characterized by acute lethal infection with analysis, amino acid sequence of the F protein cleav-
intestinal hemorrhage, velogenic neuro- tropic age site, and pathogenicity index test results revealed
(America type) characterized by respiration and nerve the NDV isolate, designated as NDV/Bali-1/07, to be
lesion without intestinal lesion with high mortal- ity, a novel Indonesia velogenic NDV strain belonging
mesogenic characterized by respiratory and nerve to Group VII [5]. Since 2011 and during 2012 highly
lesion with low mortality, and lentogenic characterized related 40 NDV isolates from subgenotype VIIi have
by asymptomatic lesion in the intestine [2,3].
been isolated from poultry production facilities and
Copyright: Etriwati, et al. Open Access. This article is distributed occasionally from pet birds, throughout Indonesia,
under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/ Pakistan, and Israel [6]. NDV pathogenicity spread-
by/4.0/), which permits unrestricted use, distribution, and ing in Indonesia today is a part of virulent strain
reproduction in any medium, provided you give appropriate credit
to the original author(s) and the source, provide a link to the (mesogenic/velogenic) [4,5,7]. Based on those report,
Creative Commons license, and indicate if changes were made. virus strain in Indonesia has been estimated
The Creative Commons Public Domain Dedication waiver (http://
creativecommons.org/publicdomain/zero/1.0/) applies to the data
underwent genetic changes. Diagnosis steps that have
made available in this article, unless otherwise stated. been estab- lished by pathologist in the field in
Veterinary World, EISSN: 2231-0916 1
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course of the disease are still relying on ND assessment was done descriptively according to the
pathogno- monic symptoms and reported lesions. presence of degener- ation and necrosis, hemorrhage,
Lesions most reported in ND infection is matting of congestion, edema,
vent feathers, petechiae hemorrhage on the tip of
proventriculus papilla, ventriculus, intestine and
cecal tonsil, conges- tion of trachea and lung, and
degeneration of ovarian follicles [4,8-10].
In field case, ND lesions are often easily con-
fused with other poultry diseases, therefore to ascer-
tain whether the tissue lesions are caused by NDV
infection or not, studies about antigen distribution is
critical in observing the existence of NDV within tis-
sue. Immunohistochemistry staining can be used to
explain pathogenesis of ND in field cases. In immu-
nohistochemistry staining, an antibody reacting to a
specific antigen is used to localize the antigen within
affected tissue, which increases the accuracy of a
diagnosis as it allows identification of antigens
within tissues [11]. This research aimed to determine
NDV distribution pattern in chicken internal organ
from field cases in West Java, Indonesia.
Materials and Methods
Ethical approval
This research has been approved by the Animal
Care and Use Committee of Research and
Community Services Institution, Bogor Agricultural
University (IPB) with approval number: 42-2017
IPB.
Research procedure
10 field cases from female and male broiler
chicken aged 3 and 4 weeks, layer chicken aged 14,
26, and 33 weeks, and female domestic chicken aged
8 and 24 weeks. All chickens were diagnosed as
infected by ND by pathologist and real-time-reverse
transcrip- tion polymerase chain reaction test of all
samples con- cluded to be positive of ND.
Histopathology organ samples were trachea, lung,
heart, proventriculus, duodenum, cecal tonsil, liver,
pancreas, kidney, and brain. All organ samples were
cut into 1 cm × 1 cm
× 0.5 cm and fixed by neutral buffered formalin 10%
solution of at least 24 h before they were then made
into paraffin block histopathological section.
Clinical symptoms
Clinical symptoms were obtained from anamne-
sis with owner which consists of depression,
anorexia, greenish-white diarrhea, torticollis, the
number mor- bidity and mortality, the ras, type, and
chicken age.
Macroscopic observation
Macroscopic changes observed from each organ
consist of abnormalities in color, shape, consistency,
degeneration and necrosis, circulatory disturbances,
inflammation, and exudation.
Microscopic observation
Tissue section that has been fixed on
microscope glass was stained by hematoxylin and
eosin (H and E). The histopathological lesion
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and inflammatory cell infiltration. The severity of however, there was high morbidity and mortality.
each organ was graded with the following criteria: Whereas in unvaccinated domestic chicken the
Focal lesion distribution (low severity), multifocal morbidity and mortality was 100%. Observed
(moder- ate severity), and diffuse (high severity). symptoms were
Observation was conducted using 100× decrease appetite, histochemistry was
magnification and 3× view field repetition. lethargy, weight loss, severe on all chicken.
Immunohistochemistry examination
and green- ish-white Lung lesion on all
diarrhea in all chicken type of chicken
Tissue section that has been attached on a grossly
micro- scopic slide using poly-L-lysine 1% was examined. In young
chicken, symptoms showed moderate
deparaf- finized, rehydrated with distilled water and severity, while
flowing with tap water for 5 min, then followed by include stunted growth
and adult hen exhibit microscopic lesion and
phosphate buffered saline (PBS) Tween [12]. immunopositive reaction
lowering egg production
Antigen retrieval process was conducted using Dako showed high severity. By
and watery egg albumin.
REAL™ tar- get retrieval solution (10×) (Dako, the age of chicken, in all
Respiratory disorder was
S203130) pH 6.0 inside a microwave in 100°C of the age chicken, lung
more prominent in
temperature for 15 min. Tissue section was moved gross lesions were on
young. Neurologic
and left for 20 min at room temperature and then moderate severity,
disorders exhibited were
washed with PBS Tween for 5 min. Endogenous whereas microscopic
paralysis and torticollis.
activity blocking was performed by immersion in lesions and
H2O2 3% at room temperature for 15 min, after Respiratory organ
immunopositive
which the sections were washed with PBS Tween. Gross trachea
reactions showed high
Nonspecific protein binding blocking was per- observation revealed
severity in every age.
formed using normal goat serum 10% (Dako, catarrhal tracheitis.
Microscopically the Circulatory organ
X0907) in room temperature for 30 min and the
trachea showed con- Heart grossly
section was washed again by PBS Tween. Primary showed myocardium
antibody rabbit anti-NDV HN protein polyclonal gestion, hemorrhage,
mononuclear degeneration. In layer
antibody (1:500 in antibody diluent, Dako, S3022) chicken, it was
was spilled over the section, incubated in room inflammatory cell
infiltration, and edema accompanied by
temperature for 1 h and washed by PBS Tween for 5 pericarditis. Microscopic
min. Sections were then dipped in secondary at almost every trachea
sam- ple. The lungs observation on all heart
antibody Dako REAL™ envi- sion™/HRP, samples found
suffered pneumonia with
rabbit/mouse (ENV) (K5007) for 30 min and epicarditis and
microscopic lesion of
washed by PBS Tween for 5 min. The Dako myocarditis marked by
congestion, edema, and
REAL™ DAB+chromogen in Dako REAL™ sub- myocardium
mononuclear inflam-
strate buffer (K5007) was applied in room degeneration, and
matory infiltration at the
temperature for 40 s then washed by flowing tap necrosis, edema, with
inside and the wall of
water for 10 min. The sections were immersed in alveoli. mononuclear cell
distilled water three turnover 5 min each. Immunohistochemistry infiltration (Figure-1A).
Counterstain used was Mayer’s hematoxylin. The staining of trachea Immunopositive reaction
degree immunopositive toward ND in each organ revealed in all heart samples was
was scored as low (1-10 immunoposi- tive cell immunopositive reaction detected in the
toward NDV), moderate (11-20 immunopos- itive at cytoplasm of mucous cytoplasm of
cell toward NDV), and high (more than 20 cells epithelial cells and myocardium,
immunopositive toward NDV). Observation was cytoplasm of inflammatory cell, and in
done using 400× magnification with 3 times macrophage at sub- vascular endo- thelial
repetition of view field. mucosal layer. cell (Figure-1E).
Statistical analysis Immunopositive Based on the type
Data obtained from macroscopic and micro- reaction in lungs was of chicken, the degree of
scopic observation were analyzed descriptively and found in cytoplasm of gross and microscopic
scored based on the lesion distribution. The parabronchial epithelial lesion of layer chickens
immuno- histochemistry result was described based cell, pneumocyte, and were more severe
on immu- nopositive location and scored according inflammatory cell of compared to broiler and
alveoli. Based on the domestic chicken which
to the rate of immunopositive cell per field of view.
type and age of the have moderate severity.
Results chicken, the degree of Immunopositive degree
Clinical symptoms lesion found in trachea was higher in broiler and
Based on anamnesis it is known that both by gross, microscopic, domestic chicken
broiler and layer chicken have been vaccinated; and immuno- compared to layer
chicken with showed
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moderate degree. Based surface and epithelial reaction was found cytoplasm of
on age, macroscopic gland, hyperemia of the mucosal epithelial cells hepatocytes (Figure-
and microscopic lesion muscu- laris layer and and inflammatory cell 1G).
degree is more severe infiltration of at the submucosal layer. Pancreas grossly
in older chicken inflammatory cells to Cecal tonsil showed multifocal
compared to young sub- mucosal layer of showed typhlitis on necrotic pancreatitis.
chicken which has proventriculus glands every sample. While microscopic
moderate severity. (Figure-1B). Epithelial Microscopic examination showed
However, mucosal cells, examination of cecal multifocal hemorrhage
immunopositive degree submucosal tonsil revealed that and inflammatory cell
was higher in young inflammatory cell, and there was necrosis of accu- mulation between
chicken compared to glandular epithelial epithelial mucosal cell, acinar cells.
older chicken. cells of proventriculus hem- orrhage, and Immunopositive reac-
Digestive organ all showed infiltration of tion was found on
Most immunopositive inflammatory cell into acinar cells and
proventriculus showed reaction toward NDV the submucosal layer. inflammatory cells.
catarrhal proven- (Figure-1F). Based on Immunopositive Based on the type of
triculitis and only the type of chicken, reaction was found at chicken, macroscopic
samples from domestic proven- triculus gross mucosal epithelial cells and microscopic lesion
chicken showed lesion degree in and inflammatory cells and the severity degree
hemorrhagic domestic chicken is at the lymphoid follicle. in chicken liver and
proventriculitis. much severe compared Macroscopic and pancreas were more
Microscopic to broiler and layer micro- scopic lesion severe in layer chicken
observation revealed chicken which only had and immunopositive compared to moderate
proventriculitis in every a moderate degree. reaction showed high degree of broiler and
sample, marked by Domestic chicken severity on every type domestic chicken.
desquamation of proventriculus showed and chicken age. Higher immunopositive
proventriculus epithelial hemorrhage while Liver showed degree of liver and
broiler and layer degeneration and pancreas was found in
chicken only showed multifocal necro- sis in broiler chicken and
swelling with catarrhal domestic chicken and domestic chicken,
exudation. Microscopic broiler chicken, while compared with layer
and in layer chicken they chicken which showed
immunohistochemis- were accompanied by moderate degree. Based
try observation showed perihepati- tis. on age difference,
high severity in all Microscopic macro- scopic and
chicken type. Based on examination revealed microscopic lesion was
age difference, hemorrhage, necrosis, more severe in old
macroscopic lesion was and multifocal chicken compared to
spread in moderate inflammatory cell young, while
degree in all of age; infiltration (Figure-1C). immunoposi- tive
however, microscopic Immunopositive reaction was higher in
and reaction was found at young compared to
immunohistochemistry the macrophage around older chicken which
lesion was distributed central vein and the showed moderate
as high severity in immunopositive.
every age.
Duodenum
suffered catarrhal
enteritis in every
sample. Microscopic
examination revealed
des- quamation and
necrosis of epithelial
mucosal cell,
hemorrhage, and
inflammatory cell
infiltration into
submucosal layer.
Immunopositive
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and mononuclear cell
infiltration (b). (G) Liver lesion and also chicken because
immunopositive reaction immunopositive respiratory tract is one
was distributed on reaction was severe in of the main gateways of
cytoplasm of hepatocytes every type and every NDV. Virus attaches to
(a), and at the
macrophage around chicken age group. the respiratory epithelial
central vein (b). (H) Brain Nervous system
cell by utilizing sialic
A B
immunopositive reaction
The brain grossly acid on host cell as
was distributed glial cell receptor [13]. The
(a), mononuclear cells of suffered encephalitis
perivascular cuffing (b), with microscopic lesion presence of virus on
and on neuron cytoplasm showing hyperemia, respiratory epithelial
(c). Hematoxylin and eosin
perivascular cuffing, cell will induce regional
staining (A, B, C, D), immunity sys- tem to
immunohistochemistry edema, gliosis, and
staining with rabbit anti- satellitosis (Figure-1D). produce specific
C D immunoglobulin A
NDV hemagglutinin- Immunopositive
neuraminidase protein
reaction was distributed antibody which will
polyclonal antibody (E, F, phagocyte virus.
G, H). on neuron cytoplasm,
glial cell, and Response observable on
mononuclear cells of the respiratory tract is
Urinary organ
peri- vascular cuffs the presence of
The kidney in
(Figure-1H). The congestion and increase
every sample grossly
E F
degree of macroscopic of mucous exudate
had nephri- tis marked
and microscopic lesion secretion into the
by swelling, fragility
and also trachea, where the body
with multinecrotic foci.
immunopositive reac- will then attempt to
Microscopic
tion was severe in every expel virus antigen from
examination revealed
type and every chicken respiratory tract,
hemorrhage, tubular
age group. protecting the epithelial
epithelial cell necrosis,
G H sur- face from virus
and inflammatory cell Discussion
Figure-1: Histopathology
infiltration in the attachment and invasion
changes and [14]. Young bird
distribution of Newcastle interstitium. The confirmation
disease virus (NDV) in Immunopositive reac- of ND suspect chicken infected by NDV
internal organs of tion was found on diagno- sis with showed more severe and
chickens from a field case
tubular epithelial cell, immunohistochemical acute clinical symptoms
in Indonesia. (A) Heart compared to older bird
microscopic observation blood vessel endothelial staining in this study
showed degeneration cells, and macrophages proves that the [2]. Lung lesion is the
(a), mononuclear cell within the glomer- ulus. immunohistochemistry result of circulatory
infiltration (b), and
The degree of method can be used in disturbance caused by
myocardium necrosis (c). viremia and bacterial
(B) Proventriculus macroscopic and diagnosing ND
microscopic observation microscopic accurately, quickly, and secondary infection
showed hyperemia on more economically than [15]. Immunopositive
glandular epithelial cells of serologically and reaction in both of the
proventriculus glands (a). respiratory organ
(C) Liver microscopic
molecularly identifies
observation showed viruses. Other matches reports by
multifocal inflammatory advantages include: (1) Nakamura et al. [16]
cell infiltration (a). (D) Can trace the that stated trachea, air
Brain microscopic sac, and lungs are
observation showed
distribution of virus in
hyperemia with various organs so it can immunopositive toward
perivascular cuffing (a), be used to know the NDV antigen.
edema (b), gliosis (c), and pathogenesis of NDV Anemia observed
satellitosis (d). (E) Heart from the carcass,
infec- tion in chickens
immunopositive reaction
and (2) safe because of especially at the breast
was detected in the
cytoplasm of myocardium the diluent of the organ muscle of all samples
(a), mononuclear cell that has been fixed with was associated with the
infiltration (c), and in formaldehyde solution hemorrhage at some
vascular endothelial cell internal organ due to
(insert) (b). (F) so that transmission to
Proventriculus sensitive host can be NDV replication [17].
immunopositive reaction avoided [11]. Macroscopic and
was distributed on Respiratory microscopic lesion
glandular epithelial cells of severity degree in layer
proventriculus glands (a), impairment was
observable in every chicken was higher
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com- pared to broiler
chicken and domestic
chicken, how- ever, the
immunohistochemistry
examination showed
only moderate
immunopositive
degree. This may be
disease which cause the
higher lesion to heart.
ND infection is usually
accompanied by
Escherichia coli which
causes perihepatitis and
fibrinous pericarditis,
and even fibrinous
purulent lesion [18].
NDV distri- bution in
heart tissue was caused
by NDV carried by
blood into the heart
(viremia), marked by
immunopos- itive
reaction to blood vessel
endothelial cell in heart
and other organ. NDV
distribution in heart
matches previous report
by Bwala et al. [19] that
stated chicken infected
by velogenic
viscerotropic isolate
showed macrophage
cell accumulation at the
myocardium and
immunopositive
reaction toward NDV is
found at myocardium
and mononuclear cells.
Based on
observation of clinical
symptoms on digestive
organ, it is known that
the ND infection was of
velogenic type. NDV
velogenic type infection
can cause more severe
infection compared to
mesogenic or lentogenic
NDV. Mesogenic NDV
infection can cause
macroscopic and
microscopic lesion;
however, the lesions
will spread as much as
velogenic virus infec-
tion [20]. NDV
replication within
intestinal lymphoid
follicle causes
Veterinary World, EISSN: 2231-0916
explained by the
presence of pericarditis
in layers, which it may
be assumed that the
more severity ero- sion
may have been caused
by the infection of
another
hemorrhage and edema
in internal organ due to
blood vessel injury [21].
NDV distri- bution in
internal organ is as
reported by Nakamura
et al. [16], Bwala et al.
[19], that field isolate of
NDV antigen was found
immunopositive in
pancreas, duo- denum,
proventriculus, and
Bursa Fabricius.
Kidney nephritis
occurs as the virus
invades through
respiratory tract then
carried by blood circula-
tion to the kidney. Virus
replicate inside mucosal
epi- thelial cell of the
upper respiratory tract
and digestive tract then
right after infection virus
spread through blood
circulation to kidney and
bone marrow, causing
secondary viremia [13].
NDV distribution in
kidney from this research
is agreed with previous
research by Nakamura et
al. [16] who found
immunopositive reaction
toward NDV in kidney
tubular epithelial cell.
Clinical symptoms
observable originating
from the nervous system
was caused by nerve cell
destruc- tion in the brain
from infection and NDV
replication. The
presence of NDV within
brain can cause vascular
and neuron damage
which will result in an
inflamma- tory
response. Inflammatory
response begun by mac-
rophrage spread in
perivascular cuffing
from which then spread
to surrounding
astrocytes and micro-
gial [22]. NDV
distribution in nervous
organ system was
identical with a previous
report [23] which stated
that encephalitis was
found in infected
chicken brain with
velogenic viscerotropic
isolate and immunopos-
itive reaction to NDV
can be located in
inflammatory cell and
astroglia.
The high incidence
of ND in vaccinated and
unvaccinated chicken in
this research showed
that vaccination program
currently conducted in
the field is not protective
enough in preventing
disease infec- tion. At
present, ND vaccination
only protects poultry by
decreasing the severity
of the disease and
lowering mortality,
however it cannot
prevent NDV
replication, especially
virulent ND [24]. The
lack of protection in
vaccines can be affected
by various factors.
Factors that affect the
lack of protection from
vaccine today is caused
by the presence of
genetic difference of
strain between virus
used in vaccines and the
virulent virus strain in
the field. The failure of
vaccination program in
chicken farm (nursery,
layer, and meat
producer) is caused by
the elevating
physiological stress in
chicken body (internal
physiological stress),
temperature fluc-
tuation, high humidity,
and faulty vaccination
pro- gram. In addition,
the ability of NDV to
break through marginal
antibody level and has
high replication speed in
chicken’s body, which
may also be among the
causes of vaccination
program failure [24,25].
Conclusion
The distribution
pattern of 10 field
samples used in this
research is still the same
with previous reported
NDV distribution
pattern which spread
systemically in chicken
internal organ. The
degree of NDV severity
was velogenic
viscerotropic,
accompanied by velo-
genic neurothropic.
Highest NDV
distribution was found
on trachea, lungs,
proventriculus,
duodenum, cecal tonsil,
kidney, and brain.
Authors’
Contributions
EW, DR, and EH
executed the
experiment, ana- lyzed,
and interpretation of
1071
 Available at www.veterinaryworld.org/Vol.10/September-
data the tissue. EW and2017/10.pdfManual. Office chicken. Vet. World,
Yamada, M., Mase, M.
SS executed the International des 7(7): 457-462.
and Imai, K. (2008)
Epizootie, New 11. Ramos-Vara, J.A. and
experiment and Brunswick. Miller, M.A. (2014)
Pathologic and
analyzed of molecu- lar immunohistochemical
2. Alexander, D.J. and When tissue antigens
studies of Newcastle
data. All authors Senne, D.A. (2008) and antibodies get
disease (ND) in broiler
interpreted and Newcasle disease. In: along: Revisting the
chickens vaccinated
Saif, Y.N., editor. technical aspects of
critically revised the immunohistochemistry-
with ND: Severe
Disease of Poultry. 12th nonpuru- lent
manuscript for ed. IOWA, USA, the red, brown, and blue
important intellectual encephalitis and
Blackwell Publishing. technique. Vet. Pathol.,
necrotizing
contents and approved p75-94. 51(1): 42-87.
pancreatitis. Vet.
the final version. 3. Miller, P.J., Decanini, 12. Erwin, Etriwati,
Pathol., 45(6): 928-
E.L. and Afonso, C.L. Gunanti, Handharyani,
933.
Acknowledgments (2010) Newcasle E. and Noviana, D.
17. Oladele, S.B., Abdu, P.,
disease: Evolutin of (2017) Changes in
The authors are Nok, A.J., Ibrahim,
genotypes and related histopathology and
N.D.G. and Esievo,
highly thankful to the diag- nostic cytokeratin AE1/AE3
K.A.N. (2008)
rec- tor, Syiah Kuala challenges. Infect. expression in skin graft
Pathogenesis of
Genet. Evol., 10(1): with different time on
University and Bogor Indonesian local cats.
Newcastle disease virus
Agricultural University 2aa6-35. Kudu 113 strains in
4. Xiao, S., Paldurai, A., Vet. World, 10(6): 662-
relation to
in Indonesian. The Nayak, B., Samuel, A.,
666.
neuraminidase produc-
authors are also 13. Wen, G., Hu, X., Zhao,
Bharoto, A.E., tion in chickens. Vet.
K., Wang, H., Zhang,
thankful to The Prajitno, T.Y., Collins,
Z., Zhang, T., Yang, J.,
Res., 2(1): 3-8.
Indonesian Ministry of P.L. and Samal, S.K. 18. Cattoli, G., Susta, L.,
Luo, Q., Zhang, R.,
Research, Technology (2012) Complete Terregino, C. and
Pan, Z., Shao, H. and
genome sequences of Brown, C. (2011)
and Higher Education Newcasle disease virus
Yu, Q. (2016)
Newcastle disease: A
for scholarship and Molecular basis for the
strains circu- lating in review of field
thermostability of
research No. chicken populations of Newcastle disease
recognition and cur-
39475/A2.1/KP/2016. Indonesia. J. Virol., rent methods of
virus. Sci. Rep., 6:
86(10): 5969-5970. laboratory detection. J.
22492.
Competing Interests 5. Adi, A.A., Astawa, Vet. Diagn. Invest.,
14. Kothlow, S. and
The authors N.M., Putra, K.S., 23(4): 637-656.
Kaspers, B. (2008) The
declare that they have Hayashi, Y. and 19. Bwala, D.G., Clift, S.,
avian respira- tory
Matsumoto, Y. (2010) Duncan, N.M.,
no competing interests. immune system. In:
Bisschop, S.P. and
Isolation and Davison, F., Kaspers, B.
References characterization of a Oludayo, F.F. (2012)
and Schat, K.A.,
pathogenic Newcastle Determination of the
editors. Avian
1. OIE, Office disease virus from a distribution of
Immunology. 1st ed.
International des natural case in lentogenic vaccine and
Academic Press
Epizootie. (2012) Indonesia. J. Vet. Med. virulent Newcastle
Elsevier, San Diego.
Newcastle Disease. Sci., 72(3): 313-319. disease virus antigen in
p273-288.
OIE Terrestrial the oviduct of SPF and
15. Lopez, A. (2012)
commercial hen using
6. Miller, P.J., Hadas, R., study on duck and Respiratory system. In:
immunohistochemistry.
Simanov, L., Lublin, chicken in Subang McGavin, M.D. and
Res. Vet. Sci., 93(1):
A., Rehmani, S.F., District. J. Ilmu Ternak Zachary, J.F., editors.
520-528.
Wajid, A., Bibi, T., Vet., 20(2): 134-147. Pathologic Basis of
20. Kommers, G.D., King,
Khan, T.A., Yaqub, T., 9. Khorrajiya, J.H., Veterinary Disease. 5th
D.J., Seal, B.C. and
Setiyaningsih, S. and Pandey, S., Ghodasara, ed. Mosby Elsivier, St.
Brown, C.C. (2001)
Afonso, C.L. (2015) P.D., Joshi, B.P., Louis. p458-538.
Virulence of pigeon-
Identification of new Prajapati, K.S., 16. Nakamura, K., Ohtsu,
origin Newcastle
sub-genotypes of Hodasara, D.J. and N., Nakamura, T.,
disease virus isolates
virulent Newcastle Mathakiya, R.A. (2015) Yamamoto, Y.,
for domestic chickens.
disease virus with Patho-epidemiological Avian. Dis., 45(4): 906-
potential panzootic study on genotype-XIII 921.
features. Infect. Genet. Newcastle disease virus 21. Eze, C.P., Okoye,
Evol., 29(1): 216-229. infection in commercial J.O.A., Ogbonna, I.O.,
7. Darniati, S.S. and vaccinated layer farms. Ezema, W.S., Eze,
Indrawati, A. (2015) Vet. World, 8(3): 372- D.C., Okwor, E.C., Ibu,
Molecular detec- tion 381. J.O. and Salihu, E.A.
and diversity of 10. Balachandran, P., (2014) Comparative
Newcastle disease virus Srinivasan, P., study of the pathology
isolates from native Sivaseelan, S., and pathogenesis of a
chickens in Aceh. J. Balasubramaniam, G.A. local velogenic
Ked. Hewan, 9(2): 178- and Gopala Krishna Newcastle disease virus
184. Murthy, T.R. (2014) infection in ducks and
8. Panus, A., Isolation and chickens. Int. J. Poult.
Setiyaningsih, S. and characterization of Sci., 13(1): 52-61.
Mayasari, N.L.P. Newcastle dis- ease 22. Zachary, J.F. (2012)
(2015) Newcastle virus from vaccinated Nervous system. In:
disease virus infection commercial layer
Veterinary World, EISSN: 2231-0916 1072
 Available at www.veterinaryworld.org/Vol.10/September-
McGavin, M.D. and2017/10.pdfMiller, P.J. (2013)
Zachary, J.F., editors. Immune responses of
Pathologic Basis of poultry to Newcastle
Veterinary Disease. 5th disease virus. Dev.
ed. Mosby Elsivier, St. Comp. Immunol.,
Louis. p771-870. 41(3): 447-453.
23. Ecco, R., Susta, L., 25. Wang, X., Zhou, Q.,
Afonso, C.L., Miller, Shen, J., Yao, J. and
P.J. and Brown, C. Yang, X. (2015) Effect
(2011) Neurological of difference doses of
lesins in chickens Newcastle disease
experimentally vaccine immunization
infected with virulent on growth
Newcastle disease performance, plasma
virus isolates. Avian. variables and immune
Pathol., 40(2): 145- response of broilers. J.
152. Anim. Sci. Biotechnol.,
24. Kapczynski, D.R., 6(1): 1-5.
Afonso, C.L. and

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