CLINICAL

Veterinary
REPORT Veterinary Research Forum. 2015; 6 (1) 95 - 98
Research
Forum
Journal Homepage: vrf.iranjournals.ir

Adenovirus-like inclusion body hepatitis in a flock of broiler chickens in

Kermanshah province, Iran
Morad Rahimi1* , Zahra Minoosh Siavosh Haghighi2
1
Department of Clinical Sciences, Faculty of Veterinary Medicine, Razi University, Kermanshah, Iran; 2 Department of Pathobiology, Faculty of Veterinary
Medicine, Razi University, Kermanshah, Iran.

Article Info Abstract

Article history: Inclusion body hepatitis (IBH) has been reported in many countries in the world. The IBH or
similar cases characterized by hepatitis and presence of intra-nuclear inclusion bodies in
Received: 18 July 2013 hepatocytes have not been reported in broiler chickens in Iran. This is the first report on
Accepted: 04 January 2014 outbreak of adenovirus-like inclusion body hepatitis in Iran. On October 2012, an onset of high
Available online: 15 March 2015 acute mortality in a flock of 2 day-old broiler chickens was reported to the Veterinary Clinic,
Faculty of Veterinary Medicine, Razi University, Kermanshah, Iran. The birds showed lethargy,
Key words: huddling, ruffled feathers, and inappetence. At necropsy the livers were the primary organ
affected which were enlarged, pale yellow with necrotic foci and multiple petechial
Adenovirus hemorrhages. Tissue samples of liver, kidneys and heart were fixed in 10% buffered formalin.
Broiler They were processed and stained with hematoxylin and eosin for histopathological studies.
Hepatitis Significant microscopic lesions were seen in the livers. Large eosinophilic intra-nuclear
Inclusion body inclusion bodies were seen in hepatocytes. Based on the acute high mortality, age of the
Iran broilers, gross lesions and histopathological findings (especially intra-nuclear inclusion bodies),
the condition was diagnosed as adenovirus-like inclusion body hepatitis.
© 2015 Urmia University. All rights reserved.

‫هپاتیت شبیه به هپاتیت آدنوویروسی همراه با گنجیدگی در یک گله مرغ گوشتی در استان کرمانشاه‬
‫چکیده‬
،‫ چون تاکنون هپاتیت همراه با گنجیدگی یا موارد مشابه که مشخصه آن وجود گنجیدگی در هپاتوسیت ها باشد‬.‫بروز هپاتیت همراه با گنجیدگی در بسیاری از کشورهای جهان گزارش شده است‬
‫ در یک گله مرغ گوشتی‬،‫ شبیه به هپاتیت دارای گنجدیدگی داخل سلولی آدنوویروسی‬،‫ در این گزارش بروز موردی از هپاتیت همراه با گنجیدگی‬،‫در گله های مرغ گوشتی ایران گزارش نشده است‬
‫ طی‬.‫ روزگی به کلینیک دانشکده دامپزشکی دانشگاه رازی گزارش شد‬2 ‫ وقوع مرگ و میر حاد به میزان باال در یک گله مرغ گوشتی در سن‬1931 ‫ در مهر ماه‬.‫در ایران برای اولین بار گزارش می شود‬
‫ کبد اندام اصلی تحت تأثیر بیماری بود که‬،‫ در کالبدگشایی‬.‫ ژولیدگی پرها و بی اشتهایی در گله بیمار مشاهده شد‬،‫ انباشته شدن روی هم‬،‫ عالئمی از قبیل بیحالی‬،‫بازدید مکرر از مزرعه و معاینات بالینی‬
‫ برش های بافتی با‬،‫ درصد بافری قرار داده شد و پس از فرآوری‬11 ‫ کلیه ها و قلب در فرمالین‬،‫ نمونه های بافتی از کبد‬.‫ همراه با کانون های نکروتیک و خونریزی های پتشی بود‬،‫ زرد کم رنگ‬،‫بزرگ‬
‫ به ویژه گنجیدگی های داخل هسته ای بزرگ و ائوزینوفیلیک در هپاتوسیت ها‬،‫ ضایعات عمده میکروسکوپی در کبد‬.‫هماتوکسیلین و ائوزین رنگ آمیزی و مورد مطالعه هیستوپاتولوژی قرار گرفتند‬
‫ وضعیت‬،‫ به خصوص گنجیدگی های داخل هسته ای‬،‫ جراحات ماکروسکوپی و یافته های هیستوپاتولوژی‬،‫ مرگ و میر حاد به میزان باال‬،‫ بر اساس سن جوجه های گوشتی درگیر بیماری‬.‫مشاهده شدند‬
.‫مشاهده شده در گله مرغ گوشتی مبتال به عنوان هپاتیت شبیه به هپاتیت آدنوویروسی همراه با گنجیدگی تشخیص داده شد‬

‫ هپاتیت‬،‫ مرغ گوشتی‬،‫ گنجیدگی داخل سلولی‬،‫ ایران‬،‫ آدنوویروس‬:‫واژه های کلیدی‬

*Correspondence:
Morad Rahimi. DVM, PhD
Department of Clinical Sciences, Faculty of Veterinary Medicine, Razi University, Kermanshah, Iran
E-mail: [email protected]
96 M. Rahimi and Z. Minoosh Siavosh Haghighi. Veterinary Research Forum. 2015; 6 (1) 95 - 98

Introduction University, Kermanshah, Iran. During the disease

outbreak, the farm was visited regularly. The farm was
Inclusion body hepatitis (IBH) was recognized for the located in northwest of Kermanshah province. Totally,
first time in the USA by Helmboldt and Frazier.1 Since then, 35,000 broiler chickens were kept in 5 windowless houses
the disease has been reported in many countries in the (7000 birds per house). Wood shavings were used as litter.
world. The IBH is caused by several serotypes of fowl All houses were equipped with central heaters, pad cooling
adenovirus.2,3 Fowl adenovirus are resistant to several systems and ventilators to provide the optimum
disinfectants, heat and pH changes.4 Although IBH is environmental conditions for the broiler chickens. The
transmitted by both vertical and horizontal means, vertical birds were fed with a commercial corn-soybean meal
transmission is reported as a very efficient way to spread starter diet. Feed and water were provided ad libitum. The
from parent birds to progenies.5,6 Horizontal transmission lighting program was 23 hr light and 1 hr dark. All
of the causative virus occurs by the oral-fecal route and chickens of the farm were vaccinated against infectious
further spread takes place by mechanical means and by bronchitis virus at the time of arrival to the farm.
contamination with infected feces.4 At farm visits, clinical signs and daily mortality were
Inclusion body hepatitis normally occurs in meat-type recorded. Clinically, the birds showed lethargy, ruffled
chickens at 3 to 7 weeks of age, but it has been reported in feathers, and inappetence. Huddling and smothering were
birds as young as 7 day-old and as old as 20 weeks. 7,8 In the major observed clinical signs. Mortality of the disease
natural outbreaks, the disease is characterized by sudden started with 300 chicks per 24 hr on 2nd day of age, peaked
onset of mortality ranging from 2 to 40 percent in at 410 on 5th day and then gradually declined to normal at
chickens. High mortality occurs when the affected birds the age of 21 days. During the course of disease, total
are less than three weeks of age. Depending on the mortality reached about 14%.
pathogenicity of the virus, immune status of the chicks and On daily visits, a complete necropsy was carried out on
concurrent secondary infections, mortality up to 80% has at least five newly dead or euthanized sick birds of each
been reported. Generally, mortality peaks within three to house. Tissue samples of liver, kidneys and heart were
four days and declines within 9-14 days. Clinically, affected fixed in 10% buffered formalin. They were processed
birds show lethargy, huddling, ruffled feathers, and according to routine procedures and stained with
inappetence.4 Gross lesions of IBH include an enlarged hematoxylin and eosin (H & E) for histopathology.
pale and friable liver sometimes with necrotic foci. Due to some restrictions, we could not follow up the
Ecchymotic hemorrhages may be also seen in the liver and case with virus isolation or molecular methods to identify
less consistently in leg and breast muscles. In most cases, the exact etiological agent of the disease. Therefore,
the main lesions are in the liver.5,9,10 based on the acute high mortality, age of the broilers, and
The laboratory diagnosis of adenovirus infections, gross and microscopic lesions (especially intra-nuclear
including IBH in chickens is in most cases based on inclusion bodies), the condition was diagnosed as
histological investigations and detection of intra-nuclear adenovirus-like inclusion body hepatitis. At necropsy the
inclusion bodies in hepatocytes or detection of the antigen livers were the primary organ affected which were
or virus particles using immunofluorescence test or enlarged, pale yellow with necrotic foci and multiple
electron microscopy.4 Histopathological lesions include petechial hemorrhages (Fig. 1).
areas of focal necrosis and some of the hepatocytes have
intra-nuclear inclusion bodies. Inclusion bodies can be
eosinophilic, large, round, or irregularly shaped with a clear
pale halo or occasionally basophilic.11-13 Sub-epicardial
hemorrhages with multifocal necrosis in the myocardium
have been the major reported findings in the heart.14,15
According to our current knowledge, IBH or similar
cases characterized by hepatitis and presence of intra-
nuclear inclusion bodies in hepatocytes have not been
reported in broiler chickens in Iran. Therefore, this
outbreak of adenovirus-like inclusion body hepatitis in a
broiler farm in Iran is reported here for the first time.

Case Description

On October 2012, an onset of high acute mortality in Fig. 1. Chicken livers affected by adenovirus-like inclusion body
a flock of 2-day old broiler chickens was reported to hepatitis; they were enlarged, pale yellow and friable with
Veterinary Clinic, Faculty of Veterinary Medicine, Razi necrotic foci and petechial hemorrhages.
 M. Rahimi and Z. Minoosh Siavosh Haghighi. Veterinary Research Forum. 2015; 6 (1) 95 - 98 97

Kidneys appeared swollen and pale (Fig. 2). On effect with avian adenoviruses, as the age of the host
histopathological examinations, significant lesions were increases, the degree of multiplication of the viruses
seen in the liver. Focal hepatitis with infiltration of within the host is restricted16 and the mortality they
mononuclear inflammatory cells was noted. Large produce is reduced.17
eosinophilic intra-nuclear inclusion bodies were seen in Some of the clinical signs and lesions of the disease
hepatocytes (Fig. 3). Kidneys showed severe hyperemia, reported here were similar to those of hydropericardium
tubular epithelial cells degeneration, intra-tubular cellular syndrome (HPS; Angara disease) reported by Ahmad et al.
cast formation, and mild interstitial nephritis. Mild focal and Anjum et al. 14,18 However, the course of the disease,
myocarditis, with degenerated muscle fibers was seen in mortality rate and other lesions were different. It started
the heart sections. at two days of age, the mortality rate (14%) was lower
than that of HPS, and there was no accumulation of straw
colored fluid in the pericardial sac. Typically, the mortality
of HPS starts at three weeks of age, and it causes between
20 and 80% mortality.18,19 The main pathological findings
of HPS are the accumulation of a clear, straw colored fluid
in the pericardial sac.20
We could not follow up the case with virus isolation
or molecular methods to identify the exact etiological
agent of the disease. Therefore, based on the acute high
mortality, age of the broilers, and gross and microscopic
lesions (especially intra-nuclear inclusion bodies), the
condition was diagnosed as adenovirus-like inclusion
body hepatitis. Virus isolation can be performed on
cloacal swabs (feces) and fresh liver with lesions. The
isolation of an adenovirus from the appropriate organ
does not necessarily mean it is the etiological agent of the
Fig. 2. Swelling (Arrow) and urates deposition in the kidneys of a
disease. Such an isolate may be the causative agent, it
broiler chicken with adenovirus-like inclusion body hepatitis.
may have been latent in the bird or it may have been a
chance isolate. In assessing the potential of an isolate to
cause disease the immune status of the chick must be
considered.5 Pathognomonic intra-nuclear inclusion
bodies in liver cells were the same as IBH of avian
species. Hematoxylin and eosin staining of liver sections,
and demonstration of intra-nuclear eosinophilic
inclusions, may provide a nonspecific indication of the
presence of DNA containing virus.
Further studies have to be done to assess the
prevalence of serotypes of fowl adenoviruses and
occurrence of IBH in poultry flocks in Iran. Demonstration
of pathognomonic intra-nuclear inclusion bodies in
hepatocytes indicates the involvement of a DNA containing
virus. Serology can be used to monitor progression but
obviously does not indicate active infection. Virus isolation
Fig. 3. Liver, large eosinophilic intra-nuclear inclusion body and molecular methods can also be used for detection and
(Arrow), (H & E, 1000×). typing of field isolates.

Discussion References

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