Parentage verification of Valle del Belice dairy sheep using microsatellite markers

Abstract

The objectives of the present work were to evaluate polymorphism of microsatellite markers, develop a parentage test and estimate misidentification rate on Valle del Belice Sicilian dairy sheep breed. Polymorphism was evaluated from 184 randomly selected animals from 5 different flocks. A total of 21 microsatellite markers including CSRD247, ILST011, McM527, APO010, INRA132, DU194351, DU323541, FCB128, McM16, OarCP49, ILST087, DU223896, ILST005, TCRVB06, SPS113A, INRA063A, SPS115, DU206192, DU216028, BM827 and McM54 were amplified using three PCR reactions and fragment lengths determined in a single multiplex electrophoresis run. Markers were easily genotyped, very polymorphic and in Hardy-Weinberg equilibrium (except McM527 and ILST_005) thus suitable for paternity test. Statistical analysis was performed using Cervus 3.0. The mean number of alleles per locus = 9.29, expected heterozygosity = 0.749 and PIC= 0.70. The combined non-exclusion probability = 5.5 x10-5, 5 x10-8 and 3.2 x10-13 for the first, second and the parent pair respectively. The paternity test was performed on 64 families generated with multiple sires mating. The results indicated errors in 20.3% and 28.7% of mothers and fathers respectively. In 8 cases, out of 15 maternal exclusions, the real mother was identified among the other females. Misidentification rate indicates the necessity of keeping more efficient control of genealogical records.