Development of 12 novel polymorphic microsatellite markers using a next generation sequencing approach for Spiculopteragia spiculoptera, a nematode parasite of deer.

ABSTRACT

Twelve novel polymorphic microsatellite markers were produced and characterized for Spiculopteragia spiculoptera (Nematoda, Trichostrongyloidae) a common parasite of abomasum of Roe and Red deer, using next generation sequencing approach, and two multiplexes PCR were developed with these markers. Polymorphism of each locus was tested in 40 individuals of this species from diverse wild populations of cervids, and was tested for crossed-amplification on four other species of nematodes, close to S. spiculoptera among the Trichostrongyloidea 20 Spiculopteragia houdemeri, 34 Ostertagia leptospicularis, 16 Ashworthius sidemi, and 25 Trichostrongylus spp. Our new microsatellite markers seem to be specific to Spiculopteragia spiculoptera since no amplifications were obtained for the four other species. The number of alleles per locus ranged from 2 to 12, the average observed and expected heterozygosity per locus ranged from 0.025 to 0.641 and from 0.049 to 0.664, respectively. Four of the 12 microsatellite loci showed significant deviations from Hardy-Weinberg equilibrium (which two slightly significant). One locus pair showed significant linkage disequilibrium (Sspi4 vs. Sspi8). Neither evidence of scoring error due to stuttering nor evidence of large allele dropout was found at all of the 12 loci, but evidence of null alleles was indicated at three loci because of general excess of homozygotes for most allele size classes. These polymorphic loci will be useful markers to study population genetics structure of Spiculopteragia spiculoptera in order to understand transfer and to explain the relationships between deer populations.