Development and characterization of microsatellite loci in the entocommensal Malacobdella arrokeana (Nemertea: Bdellonemertea), from Patagonia (Argentina) and cross-amplification in 34 nemertean species

Microsatellite loci (26 in total) were isolated for the first time for the entocommensal nemertean Malacobdella arrokeana, using 454 GS-FLX Titanium pyrosequencing. We developed conditions for amplifying these markers in 8 multiplex and 5 individual reactions. One to 14 alleles were detected per locus across 25 samples analyzed from San Matias Gulf, Patagonia (Argentina). For the 25 polymorphic loci, observed and expected heterozygosities ranged from 0.125 to 0.920 and 0.119 to 0.890, respectively; five loci deviated from Hardy–Weinberg equilibrium. Limited distribution and host specificity of M. arrokeana, which only inhabits Panopea abbreviata, have endangered this nemertean given the unregulated commercial exploitation of its host. These useful markers provide data for future conservation strategies. Cross-amplifications were also tested in 34 nemerteans species, representing the major lineages in this phylum. In total, 18 of the 26 loci (from one to 11 per species) gave clear allelic profiles.