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Little is known about the genetic structure of microscopic animals from mosses and lichens. A few studies have investigated the geographic variation in tardigrades from mosses, but so far no study has investigated the intra-population or local clonal lineage variation. Echiniscus testudo (Echiniscoidea: Echiniscidae) belongs to a large cosmopolitan genus of terrestrial tardigrades comprising more than 150 species. It is a common tardigrade in mosses in the temperate part of the Northern hemisphere, and is highly tolerant of desiccation and freezing. In a previous study, we reported a maximum of 1.28% sequence variation (uncorrected p-distance) in cytochrome c oxidase subunit I (COI) haplotypes between clonal lineages covering a large geographical area. However, in this previous study we used pooled specimens to constitute a sample, and the genetic diversity from single specimens within a locality therefore remains unknown. Accordingly, the present study investigates the COI sequence variation and haplotype diversity between single specimens of E. testudo collected at three Danish localities, separated by 80 m and 186 km. A total of 10 COI haplotypes were found in the present study (Et2, Et3, Et9, Et12-Et18); only three of these were previously reported (Et2, Et3 and Et9). The uncorrected COI sequence diversity ranged between 0-2.07%, with haplotype Et18 having the highest genetic difference. The second most variable haplotypes (Et14, Et15, and Et17) all showed a maximum diversity of 1.19% compared to the other haplotypes. No general pattern of haplotype distribution was evident. Our data suggest that E. testudo has dispersed across the Baltic sea as haplotypes Et3, Et13 and Et14 are present at all three localities. The most likely dispersal mode is passive wind dispersal in the cryptobiotic tun stage. The current study emphasises that numerous sequences from single specimens are needed to describe the genetic diversity within single moss cushions.
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