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20031
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Subject: Veterinary Public Health × End date: 2006 × Start date: 2000 ×
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    ThesisItemOpen Access
    STUDIES ON WILDLIFE FORENSICS TO DETECT SPECIES ORIGIN OF SAMBAR (Cervus unicolor) MEAT
    (SRI VENKATESWARA VETERINARY UNIVERSITY , TIRUPATI – 517 502 (A.P) INDIA, 2003-06) VENKAT RAGAVAN, K; UMAMAHESWARA RAO, S (Major); MASTHAN REDDY, P; SANKARA REDDY, I
    ABSTRACT : Attempts were made to develop suitable wildlife forensic tools with immunologic basis to detect and identify sambar (Cervus unicolor) meat in the experimentally adulterated meat samples of domestic animal species viz., buffalo, cattle, sheep, goat, pig and rabbit both fresh and cooked (thermostable muscle protein (TMP)) meat. PBS, pH 7.3 was found to be a suitable diluent for preparation of antigens both from fresh and cooked meat of sambar, buffalo, cattle, sheep, goat, pig and rabbit. Two kinds of experimental animals viz. rabbit and sheep were immunized with sambar muscle protein antigen both fresh and cooked (TMP) to study their ability to develop specific antibody. The rabbit, a distinct phylogenetic animal was found to be more suitable when compared to sheep which is a phylogenetically related species as evidenced by the immunocompetence of the sera to recognize and react with TMP antigens leading to the formation of precipitation reaction. The sambar flesh, the source of antigen for the current investigation was collected immediately from the naturally dead sambar (either by natural death or by accidental death without any infectious etiology) from Sri Venkateswara Zoological Park, Tirupati. The conventional immunochemical methods like DID and CIE tests were used to detect wildlife sambar meat, both fresh and cooked (TMP) muscle antigens, were standardized using hyperimmune sera raised in both, rabbit and sheep. The results of precipitation reaction were quick within 6 hours of incubation for fresh muscle antigens with rabbit and sheep antisera, while the precipitation reaction was delayed by 72 hours for TMP antigens with rabbit antisera only but the reaction was absent with sheep antisera. The protein profile of sambar fresh and TMP antigens was studied by subjecting to SDS – PAGE and a comparison was made with those of other domestic animal species. The electrophoretogram revealed that sambar fresh muscle protein was resolved into seven protein bands with molecular weights of 60kd, 52kd, 44kd, 42kd, 30kd, 22kd and 12kd, while the buffalo muscle protein profile resolved into five protein bands with molecular weights of 60kd, 52kd, 44kd, 42kd and 22kd. The cattle meat profile was resolved into three protein bands: 40kd, 30kd and 12kd, while the sheep meat protein profile resolved into six bands: 60kd, 52kd, 44kd, 22kd and that of goat meat protein profile was resolved into five protein bands: 60kd, 52kd, 44kd, 42kd and 12kd. The electrophoretogram of TMP antigens of sambar and other species of domestic animals like buffalo, sheep, goat and pig except cattle showed a partially denatured two protein complex with diffused migration pattern (44kd and 42kd). The two low molecular weight protein bands 22kd and 12kd were present only in sambar probably species specific. The electroblot immuno assay (western blot) technique could identify serologically related TMP antigens of 44kd and 42kd of sambar, buffalo, sheep, goat and pig. Another serologically related protein 40kd was noticed only in cattle although not appreciated very well in sambar. Two serologically identified proteins with molecular weight of 22kd and 12kd were present only in sambar, possibly species specific. The tests like DID and CIE were carried out using rabbit or sheep antiserum to detect adulteration of sambar fresh meat from binary mixtures of fresh meat of buffalo, cattle, sheep, goat, rabbit and pig. These tests could detect the fresh meat adulteration up to 1% level while detection of sambar cooked meat (TMP) adulterated with cooked meats (TMP) of domestic animals could be up to a level of 10% only with rabbit antisera. The sheep antisera against TMP antigen of sambar did not detect the adulteration. xxi Western blotting could identify cooked (TMP) sambar meat adulterated with the cooked (TMP) meats of buffalo, cattle, sheep, goat and pig as low as up to a level of 1%. The experimental adulteration of cooked (TMP) sambar meat with cooked buffalo meat when subjected to western blotting, the detection was as low as 0.1% but not at 0.05% level.