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DETECTION AND IDENTIFICATION OF TUBERCULOSIS IN BOVINE

KrishiKosh

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Title DETECTION AND IDENTIFICATION OF TUBERCULOSIS IN BOVINE
 
Creator BASSESSAR, VARUN
 
Contributor Shrivastav, Dr. A.B
 
Subject Mycobacterium bovis
tuberculosis
 
Description Mycobacterium bovis infection has been confirmed in a wide range of mammal hosts throughout the world. The confirmatory diagnosis of tuberculosis sometimes requires several weeks, after collection of suspected tissues. It was envisaged that a well planned study to compare all the diagnostic methods, incorporating the novel molecular diagnostic tools will be beneficial in formulating an early accurate diagnosis of the infection and also provide important links in establishing epidemiology of M. bovis infection in wild and domestic animals.
Clinical examination of 125 animals, between the age group of 1-10 years, 50 at Livestock farm Adhartal, Veterinary College Jabalpur and 75 animals at Dayadoya was done to observe lymphadenopathies, loss of muscle mass, and/or production loss, intermittent pyrexia, udder infection and dry cough. From these animals 10 ml. of blood was collected aseptically from jugular vein and faecal pinch was collected from rectum in 100 animals only taking care to include both clinically healthy and unhealthy animals showing respiratory distress. Samples were also collected from the carcasses of cattle that were subjected to necropsy at Livestock farm Adhartal and organised dairy farms in and around Jabalpur and in the Department of Veterinary Pathology., Veterinary College Jabalpur. .
Tuberculin testing was done in all the 100 animals, and 36 animals (36%) were found positive with an increase of 4.00 mm or more in skin fold thickness at 72 hours. The tests performed using the blood samples comprised Rapid diagnostic assay, M. bovis antibody enzyme linked immunosorbent assay and M. bovis gamma interferon ELISA for cattle.
Out of the total 100 animals screened, 31 animals (31%) were positive for the M. bovis antibody in serum by using antigen Rapid bovine TB test kit, 30 (30%) animals were positive for the M. bovis antibody in serum by using M. bovis antibody ELISA, 22 (22%) animals were positive for bovine tuberculin and 2 (2%) animals were positive for avian tuberculin Mycobacterium bovis gamma interferon ELISA for Cattle
Polymerase amplification assay (PCR) by using IS6110 primers was performed on 100 faecal & blood samples. Out of the total 100 faecal samples examined, 39 faecal samples were positive for the insertion sequence IS6110 of Mycobacterium tuberculosis complex whereas only four blood samples were positive from the total of 100 samples examined. PCR was also done on the faecal and samples of the 100 animals by using Mycobacterium tuberculosis/bovis complex PCR kit. A total of 12 (12%) faecal samples were positive and all the blood samples were negative
The faecal samples from all 100 animals were processed for isolation and cultivation of Mycobacterium spp. in the laboratory by using the standard Modified Petroff’s method. Out of the total 100 faecal samples, only 12 samples produced any visible colonies on the L-J medium with pyruvate and two in L-J medium with glycerol after 8 weeks of incubation at 370C.It was confirmed by the morphological characteristics and biochemical tests that the bacterial isolates contained 12 colonies of M. bovis and 2 colonies of M. tuberculosis. In impression smears stained by Ziehl Neelsen ( ZN) stain a total of 22 (22%) faecal samples were positive for the presence of acid fast bacilli out of the 100 animals screened.
During the study period, 12 necropsy cattle were found to have lesions suggestive for pulmonary tuberculosis. The impression smears of the affected tissue from these were stained by ZN stain and 9 were positive for acid fast staining. All the samples were positive by PCR for the Mycobacterium tuberculosis complex using IS6110 sequence and eight cases were found to be positive through Mycobacterium tuberculosis/bovis kit. Faecal sample was also collected from these cases and from the 9 cases found positive for acid fast bacilli initially, in only 4 cases acid fast bacilli could be demonstrated in the faecal smear. However, the faecal samples of 11 cases were determined as positive by using IS6110 sequence.
The results suggest that after initial screening of animals by tuberculin and gamma interferon ELISA for cattle, PCR for the Mycobacterium tuberculosis complex using IS6110 sequence can be done for early diagnosis and segregation of tuberculous animals.
 
Date 2016-12-22T11:07:23Z
2016-12-22T11:07:23Z
2013
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/92085
 
Language en
 
Format application/pdf
 
Publisher Nanaji Deshmukh Veterinary Science University, Jabalpur (M.P.)