Research Article
Standardize Polymerase Chain Reaction (PCR) Technique for the Detection of Pathogenic Serovars of Mycobacterium a. avium Infection in Layer Chicken

M.N. Haque, U.K. Rima, M.Z. Hossain, M.S. Islam, S.M.Z.H. Chowdhury, M.M. Hossain and M.A.H.N.A. Khan

Asian Journal of Poultry Science, 2016, 10(4), 175-183.

Abstract

Objectives: Avian tuberculosis (ATB) is caused by Mycobacterium a. avium, potentially zoonotic and requires adapting molecular techniques to detect pathogenic serovars and prevent zoonosis. Methodology: Layer chicken (N = 2000) of organized poultry farms of Mymensingh district showed clinical signs of progressive emaciation and reduced weight gain constituted the study materials. This study used necropsy, histopathology and Ziehl Neelsen staining to identify specific pathology of ATB in chicken. This study adapted a Polymerase Chain Reaction (PCR) technique to detect ATB in layer chicken due to highly pathogenic variant (serovars 1, 2 and 3) of Mycobacterium a. avium. Results: Investigation of sick birds at necropsy showed granulomas in liver, spleen and intestine and suspected as a case of ATB. Using histopathology, multi-focal accumulation of macrophages, epitheloid cell and lymphocytes were seen in liver, spleen, kidney, heart and intestine. Acid fast bacterium was detected in tissue sections of spleen, liver and intestine using Ziehl Neelsen staining but unable to differentiate infectivity due to pathogenic, low pathogenic and saprophytic variants of Mycobacterium. Visceral organs were, therefore, collected for PCR detection of specific cause of ATB. A specific PCR protocol was adapted targetting 16S rRNA gene (192 bp) and successfully detected pathogenic variant of ATB (M. a. avium) in clinically infected and carrier chickens. Conclusion: The PCR technique showed the potentiality to diagnosis pathogenic variant of ATB in a few hours with high degree of sensitivity and specificity. Pathogenic variant (serovars 1, 2 and 3) of ATB is highly contagious and potentially zoonotic. The PCR technique can be used to screen elderly layer chickens, diagnose ATV at early onset and dispose the infected flock to prevent future zoonosis.

ASCI-ID: 27-148

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