Arshad, Siti Suri

Universiti Putra Malaysia Institutional Repository

UPM Research Report 1997-2000, Vol II, Section 2-Extended AbstractsImmunohistochemistry detection of Infectious Bronchitis virus in chicken tissuesSftl Sufi Al'5hatJFaculty of Veterinary MedicineUniversiti Putra Malaysia43400 UPM, Serdang, SelangorMalaysiaTelephone Number of Corresponding Author: 03- 8946 8306E-mailolCorrespondingAuthor:suri@vet.upm.edu.myKey words: chicken, immunohistochemistry, kidney, IBV, tracheaIntroductionInfectious bronchitis virus (IBV) is common in chickens worldwide. IBV belongs to group 3 of the genus Coronavirus of theCoronaviridae family. It was first reported in baby chicks by Schalk and Hawn in 1931. in North Dakota, USA. Although itwas thought to be primarily a respiratory disease of chick, avian IBV has since been found to be an economically importantdisease of adult chicken. Reduced in egg production, enteritis and kidney problems have been reported associated with IBVinfection. Kidney disease, variously described as nephrosis, nephritis or uremia was first reported in the USA. In Malaysia,local nephropathogenic IBV was first noticed in 1980 and a prototype MH5365/95 was isolated in 1995. In field andexperimental infections the disease appears initially as respiratory infection followed by viremia and nephritis. Histologicalinvestigations of the development of kidney lesions throughout the course of infections have shown interstitial inflammatoryreaction and tubular degeneration. In respiratory tissues there are cilia loss and degenerative changes ofthe epithelial cells. Inthis paper. we described the use of immunohistochemistry technique to detect the location of the antigen in the kidney andrespiratory tissues of chicken infected with MH5365/95.Materials and MethodsThirty 3-weeks-old SPF chickens were infected with 0.2ml inoculum containing 10sEIDso of virus intra-tracheally. Sevenchickens served as control and received 0.2 ml PBS intra-tracheally, About 3-4 chickens and 2 chickens from the infected andcontrol groups, respectively, were killed at 3,6,9,15,18,21 and 24 days post-infection. Upon necropsy, the respiratory, kidneyand intestine tissues were collected and fixed in modified Bouin's solution and processed for standard paraffin tissue. RabbitpolycIonal antibody against MH5365/95 at the dilution of 1/50 was used as primary antibody. Immunohistochemical stainingwas performed according to the manufacturer instruction (Zymed Laboratories, South Sanfrancisco, CA).Results and DiscussionThe control chicken showed negative antigen signal in the trachea, lung, kidney and intestine. In the infected chickens, viralantigens were detected in the tracheal epithelium from day 3 to day 18. Signal was strong and distributed in mucosalepithelium of trachea and in the alveolar mucous gland and submucosa. At day 6, viral antigen was found strongly in thehyperplastic epithelial cells with strong reaction localized in the cytoplasm. In lung, strong positive antigens were detected inthe parabronchus wall, atrium, interlobular septum from day 3 to day 15 and become weak starting from day 18 onwards. Inkidney, positive signals were detected in the PCT (proximal convoluted tubule), DCT (distal convoluted tubule), CT(collecting tubule) and CD (collecting duct) of the medulla region at day 3, and the signals become more intense thereafter.The signals become less by day 15 and remained only a few in the peT and DCT. Viral antigens were also detected in therenal glomeruli, glomerular tuft, and around Bowman's capsule. The detection of viral antigens in the kidney and lung tissuesare consistent with the distribution of histological changes occurred during the course of the infection. In GIT, almost all partsof the intestine showed positive signals but signals are more intense and longer duration in lower part of the intestine namelythe duodenum, ileum, cecal tonsils and cloaca ..ConclusionsThe study enable us to understand the type of organ where the virus will persist longer. Where most of the signals in theorgans are fading away by the 18 day p.i, the ileum still showing the signals at day 21 pi. This indicate that IBV persist longerin the intestine and may suggest that the feces may served as a virus contaminant.Benefits from the studyImmunoperoxidase study enable to locate the areas in the organ where the virus replicate and persist. The signals of theantigen-antibody complexes were initially stronger and became weaker as the chicken active antibody developed during thecourse of infection.Patent(s), if applicable:NilStage of Commercialization, if applicable:218Agricultural SciencesNilProject Publications in Refereed Journals:NilProject Publications in Conference ProceedingsI. Arshad ss and Al-Salihi K. 2002. Immunohistochemistry detection of Infectious bronchitis Virus antigen in chicken respiratory and kidneytissues. In: Proceedings of the 12th Federation of Asian Veterinary Association Congress/Id" VAM Congress, 26-28 August 2002, pSIGraduate ResearchName of Research Topic Field of Degree Awarded Graduation YearGraduate ExpertiseKarima AI-Salihi Pathogenesis of mv Animal Post-doctoralin SPF chicken: bacteriologyclinical sign, grosslesion andimmunoperoxidaseLai Fu Chong Detection of Undergraduate DVM 2001infectious bronchitis projectvirus antigen ingastrointestinaltract of chicken byimmunohistochemical testIRPA Project numberO1-02-04-0305UPM Research Cluster: BAB219

Immunohistochemistry detection of infectious bronchitis virus in chicken tissues

http://psasir.upm.edu.my/42174/1/immunohistochemistry%20detection.pdf

Immunohistochemistry detection of infectious bronchitis virus in chicken tissues

Abstract

Similar works

Full text

Available Versions

Universiti Putra Malaysia Institutional Repository