A DNA fingerprinting scheme to establish the identity of semen samples of cattle breeds in artificial insemination programs of Sri Lanka.

Dairy cattle industry plays a prominent role in Sri Lankan economy because of the very high consumption of dairy milk and dairy products. Artificial Insemination is frequently used to inculcate improved performances such as high milk yield and better adaptability to the cows. However, the accuracy and outcome of Artificial Insemination is sometimes debatable due to the mixed-ups of semen samples belonged to different breeds. This could happen when semen samples are imported and distributed among regional artificial insemination centers. Hence, the availability of a robust DNA based method to identify cattle breeds at the semen stage is very important. Therefore, the present study was conducted to discriminate the five commonly used cattle breeds (Sahiwal, Jersey, Friesian, Ayrshire and Australian Friesian Sahiwal) in artificial insemination programs by using PCR based DNA fingerprinting with cattle specific microsatellite markers. The semen samples were collected from Ambewela Dairy Farms –Nuwara-Eliya, Sri Lanka (Ayrshire) and Central Artificial Insemination Station-Kundasale, Sri Lanka (Jersey, Friesian, Sahiwal, AFS). DNA was extracted from semen samples and genotypic analysis was done using 14 cattle specific microsatellite markers (ETH152, ETH225, HEL1, CSSM66, RM180, RM011, RM192, BM6425, BMS1678, BMS1941, BM3517, TGLA304, BMS1747 and ILSTS011). Only four markers generated polymorphic bands (ETH225, RM011, BM3517 and BM6425). The markers BM3517 and BM6425 can be suggested for the precise discrimination of five breeds. BM6425 generated three different bands; size ranging from 123-200 bp. Marker BM3517 displayed four different bands; size ranging from 100150 bp. In combination these two markers can be clearly used to authenticate the exact breed in which semen sample is coming and avoid any ambiguities. In addition marker BM3517 and RM011 in combination can be used to independently verify the results.