DNA testing for parentage verification in a conservation nucleus of Pantaneiro horse

Abstract

We investigated the genealogy of the in situ conservation nucleus of the Pantaneiro horse using DNA microsatellites by evaluating 101 horses, the group consisting of 71 adult horses (3 stallions, 40 male and 31 mares) and 27 foals (14 colts and 13 fillies). Genomic DNA was extracted from hair roots and genotyped using 12 microsatellite markers (AHT4, AHT5, ASB2, ASB17, ASB23, HMS3 HMS6, HMS7, HTG4, HTG10, LEX33 and VHL20). The number of alleles per locus varied from 6 to 13, with a mean of 7.8 and the expected heterozygosity ranged from 0.544 to 0.734 (mean 0.644). The VLH20, ASB2, HTG10, ASB23 markers had a high (> 0.8) polymorphism information content and the total exclusion probability of the 12 microsatellite loci was 0.99. The genealogical study of the Pantaneiro horse using genetic markers was efficient in detecting mistakes during paternity and maternity designation and is an important tool which can be used together with traditional systems of animal identification. The use of genetic markers is recommended in the systematic control of the genealogical registrations and conservation plans to improve genetic aspects of the Pantaneiro horse. The Pantaneiro horse is well adapted to the climatic conditions of the Brazilian Pantanal, a marshy region in central western Brazil in the states of Mato Grosso and Mato Grosso do Sul to which this horse is restricted. Spanish and Portuguese colonizers introduced equines into Brazil from the Iberian Peninsula, with horses arriving from Central America, across the Andes from Peru, from Northern Argentina, from Argentina or Paraguay through the Jesuit missions and directly from the Iberian Peninsula with horses imported from the Spanish region of Andalusia (Dowdall, 1982). Preservation and expansion of the Pantaneiro horse in the Brazilian Pantanal has been centered on its ability to deal with cattle and the adaptation of this breed to the Pantanal environment, this breed being preferred by farmers and ranchers in the region because it is hardy and supports the long-distance cattle drives necessary for transporting the cattle in this marshy region.Microsatellites have been used for parentage testing and individual identification in forensic science because they are highly polymorphic and show abundant sequences dispersed throughout most eukaryotic nuclear genomes (Tautz et al., 1986). Microsatellites have simple and stable inheritance when transmitted between generations and are controlled only by heredity. Furthermore, due to their small size they are efficiently amplified using the polymerase chain reaction. Many microsatellites are informative due to their high polymorphism and are useful in the paternity testing of horses (Marklund et al., 1994;Binns et al., 1995;Usha et al., 1995;Bowling et al., 1997;Tozaki et al., 2001;Cho and Cho, 2004;Lee and Cho, 2006).Our study used 12 microsatellite markers for routine DNA typing for individual identification and parentage verification of Pantaneiro horses and calculated the number of alleles (A), mean observed and ex...

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