Asian J. Med. Biol. Res. 2016, 2 (2), 177-182; doi: 10.3329/ajmbr.v2i2.29008 Asian Journal of Medical and Biological Research ISSN 2411-4472 (Print) 2412-5571 (Online) www.ebupress.com/journal/ajmbr Article Molecular detection of cattle and buffalo species meat origin using mitochondrial cytochrome b (Cyt b) gene Sirazum Munira, Fatema Tuz Jahura, Md. Munir Hossain and Mohammad Shamsul Alam Bhuiyan* Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh *Corresponding author: Professor Dr. Mohammad Shamsul Alam Bhuiyan, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh. Tel.: +88 01745748849; Fax: +88-091-61510; E-mail: msabhuiyan@gmail.com Received: 17 May 2016/Accepted: 02 June 2016/ Published: 30 June 2016 Abstract: The study was conducted to adopt PCR based technique for identification of species origin from meat samples of cattle and buffalo using mitochondrial cytochrome b (Cyt b) gene fragment. A total of 42 ear tissue and meat samples were collected from different slaughterhouses and farms of Mymensingh, Bogra and Rangpur districts and stored in 96% ethanol at room temperature. Genomic DNA was extracted from all samples using GeNet Bio genomic DNA isolation kit. The average DNA yield of considered samples was found 204.57 ng/μl where the purity ranged from 1.82–1.99. Two (2) pair species-specific primers were used to amplify Cyt b gene fragments of 472 bp and 124 bp for cattle and buffalo, respectively. The PCR results revealed different species specific amplified fragments which could discriminate between cattle (472 bp) and buffalo (124 bp) species precisely from pure and mixed samples of those species. This study suggests an accurate molecular technique for identification of cattle and buffalo species meat origin and differentiates species present in adulterated meat samples. In conclusion, this DNA based technique could be utilized for prevention of malpractice in slaughterhouse and chain shops and thereby to protect consumer’s right. Keywords: beef; buffalo meat; genomic DNA; mitochondrial Cyt b gene; PCR 1. Introduction Adulteration of food products has become a considerable problem in many countries as well as in Bangladesh by the fraudulent incorporation. Adulteration of meat may take the form of substitution of one cheaper meat with a costly expected species (Yman and Emanuelsson, 1998), for example buffalo meat (buffen) mixed with beef, which is a very old and common practice in Bangladesh. Beef is the most popular red meat in the world preferred for its taste, flavor and texture. Therefore, it is most expensive meat and consumers are ready to pay more prices for its delicacy. Whereas buffalo meat is relatively coarse and less popular compare to beef. Currently, there are about 1.46 million buffalo in Bangladesh (BER, 2015). Although a considerable number of buffalo population is available in Bangladesh and transportation of these species to the metropolitan cities is a common scenario of our country, but it is hard to find butcher shop or slaughterhouse in the market who sells buffalo meat and we found beef only. Under this circumstance, identifying the species origin of meat became a burning issue these days in Bangladesh. Different DNA-based techniques used for animal species identification which include DNA hybridization of meat samples (Lenstra et al., 2001), PCR and its variants (Matsunaga et al., 1999a and b), PCR-RFLP (Girish et al., 2005), polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) (Weder et al., 2001) and PCR-sequencing (Bartlett and Davidson, 1992). The high copy number and sequence diversity favor the utilization of mitochondrial genes over nuclear genes for species identification purposes (Kocher et al., 1989). DNA extracted from several mitochondrial genes including cytochrome b ( Cyt b) (Verma and Singh,