Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 14 3 2022 09 23 Feasibility study of genetic refinement of litter size in sheep to reduce possible fraud - A provincial study Feasibility study of genetic refinement of litter size in sheep to reduce possible fraud - A provincial study 83 100 3437 10.22103/jab.2022.19151.1391 FA Mehdi Derakhshan MSc Student, Department of Animal Sciences, Faculty of Agriculture, University of Tabriz, Tabriz, Iran. Karim Hasanpur Assistant Professor, Department of Animal Sciences, Faculty of Agriculture, University of Tabriz, Tabriz, Iran. 0000-0002-6144-9181 Arash Javanmard Assistant Professor, Department of Animal Sciences, Faculty of Agriculture, University of Tabriz, Tabriz, Iran. Jalil Shodja Professor, Department of Animal Sciences, Faculty of Agriculture, University of Tabriz, Tabriz, Iran Journal Article 2022 05 15 Objective Recently, in East Azerbaijan province, marketing of top sheep carrying fertility genes became suspected, because some farmers complained about the claim of the seller due to the low performance of such ewes on the farm. With this research motivation, the aim of the present study in the first step is to combine PCR-RFLP and subsequent PCR-Sequencing methods for genetic refinement of the FecB gene locus associated with litter size in the Booroola-Afshari sheep breed. Materials and methods In this study, overall 62 ewes and rams of Booroola-Afshari sheep and 30 ewes of Ghezel- Romanov hybrids were obtained from different sheep farms around Tabriz. Then, two-separate phases were designed, after amplification of the 190 bp region of the BMPR15 gene by PCR, two molecular PCR- RFLP methods and direct sequencing were performed according to routine laboratory instructions. Genotype results were recorded and subsequently, using different software such as POPGENE, FinchTV, and MAFFT, the genotypic, allelic, and alignment frequencies of the amplified sequences were performed to confirm the mutation, respectively. Results The results of the present report demonstrated that the source of discrepancy between the identified genotype and the actual genotype can be due to the error of reading the genotype or the seller's fraud and false claim. Also, the combination and both outputs of both molecular techniques, PCR- RFLP and direct sequencing, can minimize the technical error caused by the genotype. In addition, high reproducibility observations confirmed the presence of 8/06% fraud in the FecB gene locus genotype. Conclusions The combination of two molecular methods, PCR- RFLP and PCR-sequencing direct sequencing to detect FecB mutations, has shown acceptable efficiencies, and the establishment of government-affiliated genotyping centers for fraud detection can assist regulators in genotyping sheep with fertility genes and may reduce the number of frauds. Objective Recently, in East Azerbaijan province, marketing of top sheep carrying fertility genes became suspected, because some farmers complained about the claim of the seller due to the low performance of such ewes on the farm. With this research motivation, the aim of the present study in the first step is to combine PCR-RFLP and subsequent PCR-Sequencing methods for genetic refinement of the FecB gene locus associated with litter size in the Booroola-Afshari sheep breed. Materials and methods In this study, overall 62 ewes and rams of Booroola-Afshari sheep and 30 ewes of Ghezel- Romanov hybrids were obtained from different sheep farms around Tabriz. Then, two-separate phases were designed, after amplification of the 190 bp region of the BMPR15 gene by PCR, two molecular PCR- RFLP methods and direct sequencing were performed according to routine laboratory instructions. Genotype results were recorded and subsequently, using different software such as POPGENE, FinchTV, and MAFFT, the genotypic, allelic, and alignment frequencies of the amplified sequences were performed to confirm the mutation, respectively. Results The results of the present report demonstrated that the source of discrepancy between the identified genotype and the actual genotype can be due to the error of reading the genotype or the seller's fraud and false claim. Also, the combination and both outputs of both molecular techniques, PCR- RFLP and direct sequencing, can minimize the technical error caused by the genotype. In addition, high reproducibility observations confirmed the presence of 8/06% fraud in the FecB gene locus genotype. Conclusions The combination of two molecular methods, PCR- RFLP and PCR-sequencing direct sequencing to detect FecB mutations, has shown acceptable efficiencies, and the establishment of government-affiliated genotyping centers for fraud detection can assist regulators in genotyping sheep with fertility genes and may reduce the number of frauds.

https://jab.uk.ac.ir/article_3437_c2f32522a84d5e6357e6abac087f1b0b.pdf