46 Indian Journal of Animal Sciences 89(9): 966–969, September 2019/Article Genetic characterization of Tunisian donkey with DNA microsatellites M AROUA 1 , B JEMMALI 2 , H OULED AHMED 3 , S KHALDI 4 , S BEN SAID 5 , R BACCOUCHE 6 and M MAHOUACHI 7 Carthage University, Carthage 1054, Tunisia Received: 15 January 2019; Accepted: 20 February 2019 ABSTRACT The genetic characterization of Tunisian donkeys with microsatellite markers led to the identification of a total of 214 alleles, with an average of 14.27 alleles per locus. The number of effective alleles per locus (N e ) indicates a rich genetic polymorphism for the used markers. The number of alleles per locus (N a ) is higher than Ne,confirming the high genetic diversity in the investigated population. Observed heterozygosis(H o ) for all the studied loci inthe entire population equals 0.744. The FIS index was significantly different from zero, pointing toan excess of heterozygotes. The mean of the number of effective migrants exchanged per generation (Nm) was 3.05 indicating a high level of gene exchange. Genetic distances and the principal coordinate analysis showed three distinct population groups/breeds, with a commongenetic structure for all the individuals. This genetic characterization of the Tunisian donkey population constitutes a valuable basis for further investigations, and to the elaboration of adequate conservation strategies. Key words: Diversity, Donkey, Genetic, Microsatellites, Tunisia Present address: 1,6 PhD Scholar (arouamohamed2310 @gmail.com, baccouche.rania@gmail.com), 2,5 Associate Professor (jemmali.bayrem@gmail.com, sabensaid@gmail.com), 3,4,5 Researcher (ouledahmedh@yahoo.fr, sana.khaldi@ gmail.com), 7 Director (taymallahmah@gmail.com). The ancestor of the donkey (Equus asinus) originated from northeastern Africa. Archeological findings state that the domestication of the Nubian and Somalian wild ass (E. africanus) began in Egypt around 4000–6000 BC, and later in the Middle East, around 100 BC (Blench 2000; Beja- Pereira et al. 2004). The domestication of the donkey allowed its use for the hard physical activities, as a pack animal or for draught works (Tapsoba 2012). Nowadays, in spite of mechanization, donkey still remains a primary work stock, especially in the Saharian and mountainous regions in Africa. In Tunisia, scientists assume that the domestic donkey has evolved from the north African wild ancestor,as represented in the Roman mosaics in Sousse museum. The Tunisian donkey population is 1,23,067 distributed in the North (31.5%), Central part (51%) and South (16.5%). The Tunisian donkey breeds did not receive enough scientific attention like the other animal species with limited literature available (Denjean 1950). Several studies have reported the genetic diversity among donkey breeds using microsatellite markers (Zhang et al. 2016, Yun and Cho 2017).This study was undertaken to genetically characterize the Tunisian donkey populations with FAO recommended microsatellite markers and estimate the genetic similarity and distances among these populations. MATERIALS AND METHODS Blood samples (94) were randomly collected from Tunisian indigenous donkeys, bred in 8 governorates of Tunisia (Le Kef, Jendouba, Seliana, Beja, Kasserine, Zaghouan, La Manouba and Bizerte). The sampled animals belong to the Northern and central parts of the country. Genomic DNA was isolated from the blood sample using Invitrogen PureLink® Genomic DNA Mini-Kit, according to the manufacturer’s instructions. Genomic DNA was amplified using 15 microsatellites loci (Table 1) according to Yun and Cho (2017) and the FAO guidelines (2011). The PCR conditions were optimized with 15 min of initial denaturation at 95°C, followed by 30 cycles of 30 sec at 94°C, with the annealing temperature optimized at 58°C for 90 sec, and the extension for 1 min at 72°C. A final extension step was carried out at 60°C during 30 min to amplify the used microsatellites. The amplified products were denatured with formamide (8.3 µl) and Gene Scan-500LIZ (0.3 µl). PCR products (2 µl) were separated by capillary electrophoresis using an ABI Prism 3130® DNA genetic analyzer (Applied Biosystems™, USA), and the size analysis of DNA fragments was performed using the Gene Mapper Software® (Applied Biosystems™, Ver. 4.0). Genetic diversity within breeds, genetic variation, and relationships among breeds were assessed using different https://doi.org/10.56093/ijans.v89i9.93776