Meat speciation by molecular and serological techniques
Thumber, Jayanti M. (2002) Meat speciation by molecular and serological techniques. Masters thesis, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat, INDIA.
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Among several methods of meat speciation. Method based on DNA especially Polymerase Chain Reaction (PCR) provide potentially more information than others. The present study was under taken for meat speciation by molecular and serological techniques. Meat samples from meat species (cattle, buffalo, sheep, goat, pig and chicken) were utilized for molecular analysis. Genomic DNA was isolated from eight meat samples of each species as per method described by Sambrook et al., (1989) with some modifications. Actin multigene family was amplified by PCR using a pair of family specific primers (forward primer-5’ CCT ACA ACA GCA TCA TGA AGT G 3’and reverse primer-5’ GCT GAT CCA CATCTG CTG GAA G 3’). PCR cycling protocol included to initial denaturation at 950C for 5 minute followed by 30 cycles of 950C for1 minute (denaturation), 480C for 1 minute (annealing) , 720C for 1 minutes (extenstion). The mixture was subjected to final extension at 720C for 5 minute. PCR products were resolved by agarose gel electrophoresis it produced a characteristics band pattern for each species. PCR band pattern generated for cattle included one major band of 0.328 kb. The PCR band pattern for buffalo included two bands of 0.328 and 0.242 kb. Sheep and goat produced identical band patterns having two bands of 0.328 and 0.242 kb and one faint band of 0.765 kb. Pig also revealed pattern identical to buffalo. A band of approximately 0.330 kb size was common in all livestock species. Chicken exhibited a characteristic pattern with six bands, three bands of high intensity (approximately 0.365, 0.334 and 0.242 kb) and three faint bands (approximately o.547, 0.480 and 0.113 kb). Out of six species explored, four were found to have characteristic PCR band pattern. Thus, PCR was considered to be a potential technique for meat species identification and speciation. Actin gene band paterns were identification in both the sexes for all the species studies. PCR was also found to be consistent and effective tool as it remained same for heat treated and putrified (unpreserved) of meat. An agar gel precipitation technique was successfully used for identification of meat species, cross reaction study and detection of adulteration level in meat. Hyper immune sera were raised in rabbit by intramuscular injection of meat extract. Anti buffalo meat sera were cross-reacted to cattle, sheep and goat meat extract. At same time anti cattle meat sera were also found to be cross-reacted with buffalo, sheep and goat meat extract. Species specific sera were made by adsorption technique. These sera were used to detect level of adulteration. By AGPT, up to 10% level of adulteration between buffalo of cattle meat with sheep and goat meat was successfully detected. Conter immunoelectrohopresis techniques was also successfully used to detect meat species. This method was found to be rapid as compaired to the AGPT.
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