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We describe the development of ready-to-use set of fifteen polymorphic microsatellite markers to benefit future population biology and phylogeographic studies on the invasive Asian fish tapeworm Schyzocotyle acheilognathi . The microsatellite loci were selected from partial Illumina shotgun genome sequences of three parasite specimens and their universality tested on a set of 12 geographically distant populations of the parasite. Particularly low levels of heterozygosity have been detected in the Chinese population pointing towards possible hidden population structure that deserves further attention in future population genetic studies. [ABSTRACT FROM AUTHOR]

Dicyemida is a group of small-bodied marine parasites infecting cephalopods with many unknown life history details, such as their population structure and diversity, and their relation to sexual and asexual reproductive stages. To reveal (infra)population structure of Dicyema moschatum Whitman, 1883 in its host (Eledone moschata Lamarck, 1798), we isolated microsatellite sequences from a draft genome of D. moschatum and tested the loci for amplification success and genetic diversity. Eight microsatellite loci were selected for an analysis of D. moschatum populations from several octopus individuals sampled at two Mediterranean localities. The majority of microsatellite alleles were shared across the studied range, but several private alleles were also identified. Analysis of population structure identified two to four genetic clusters, mostly concordant with the geographic origin of the samples. Allelic patterns seen in individual dicyemid genotypes revealed that although dicyemids inside one host individual show low genetic variance, they do not represent genetically identical clones. These results suggest that infection is established by several dicyemid larvae within the lifetime of the host and sexual reproduction of dicyemids occurs inside the host. [ABSTRACT FROM AUTHOR]

The monozoic tapeworm Caryophyllaeus laticeps has been characterized by five markedly different morphotypes largely corresponding to different fish hosts. Recently, the most distinct morphotype 4 from the common nase Chondrostoma nasus was studied in more details resulting in description of a new species Caryophyllaeus chondrostomi. The molecular study based on mitochondrial cox1 and ribosomal lsrDNA did not reveal any interspecific differences between C. laticeps and C. chondrostomi and did not provide any molecular support for recognition of these two species. In the current study, six polymorphic microsatellite markers were applied in order to detect molecular differences between the two species and to provide molecular evidence of validity of C. chondrostomi. While all six microsatellite loci were amplified in different geographic populations of C. laticeps, only two of them provided the amplification product in C. chondrostomi. Results on the Bayesian analysis assigned C. chondrostomi and all geographic populations of C. laticeps to distinct clusters. Neither any close relationships among C. laticeps populations nor specific position of C. chondrostomi were revealed. Contrary, the results of the principal coordinate analysis revealed striking genetic separation of C. chondrostomi with no overlaps with any of the C. laticeps population or morphotype. Caryophyllaeus chondrostomi very probably underwent morphological divergence as a result of ongoing speciation, but this process has not yet been accompanied by sufficient genetic divergence. In this particular case, microsatellites were proved to be better molecular discriminative markers than rDNA and mtDNA. [ABSTRACT FROM AUTHOR]

Background: Fascioloides magna (Trematoda: Fasciolidae) is an important liver parasite of a wide range of free-living and domestic ruminants; it represents a remarkable species due to its large spatial distribution, invasive character, and potential to colonize new territories. The present study provides patterns of population genetic structure and admixture in F. magna across all enzootic regions in North America and natural foci in Europe, and infers migratory routes of the parasite on both continents. Methods: In total, 432 individuals from five North American enzootic regions and three European foci were analysed by 11 microsatellite loci. Genetic data were evaluated by several statistical approaches: (i) the population genetic structure of F. magna was inferred using program STRUCTURE; (ii) the genetic interrelationships between populations were analysed by PRINCIPAL COORDINATES ANALYSIS; and (iii) historical dispersal routes in North America and recent invasion routes in Europe were explored using MIGRATE. Results: The analysis of dispersal routes of the parasite in North America revealed west-east and south-north lineages that partially overlapped in the central part of the continent, where different host populations historically met. The exact origin of European populations of F. magna and their potential translocation routes were determined. Flukes from the first European focus, Italy, were related to F. magna from northern Pacific coast, while parasites from the Czech focus originated from south-eastern USA, particularly South Carolina. The Danube floodplain forests (third and still expanding focus) did not display relationship with any North American population; instead the Czech origin of the Danube population was indicated. A serial dilution of genetic diversity along the dispersion route across central and eastern Europe was observed. The results of microsatellite analyses were compared to previously acquired outputs from mitochondrial haplotype data and correlated with past human-directed translocations and natural migration of the final cervid hosts of F. magna. Conclusions: The present study revealed a complex picture of the population genetic structure and interrelationships of North American and European populations, global distribution and migratory routes of F. magna and an origin of European foci. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Microsatellite markers designed for Dibothriocephalus latus. • 122 candidate loci selected after microsatellite library screening using NGS. • 78 loci approved after PCR tests and Sanger sequencing. • 6 loci for future application in population genetic studies of D. latus. Abstract The broad fish tapeworm Dibothriocephalus latus is a causative agent of human food-borne disease called diphyllobothriosis. Medical importance, scattered geographical distribution and unknown origin of D. latus in Europe and North America make this species to be an interesting model for population genetics. Microsatellite markers were originally designed by library screening using NGS approach and validated as tools for future studies on population genetics of D. latus. Out of 122 candidates selected after NGS analysis, 110 yielded PCR products of the expected size, and in 78 of them, a declared repetitive motif was confirmed by Sanger sequencing. After the fragment analysis, six loci were proved to be polymorphic and tested for observed (Ho) and expected (He) heterozygosity, and deviations from Hardy-Weinberg equilibrium (HWE). They promise future application in studies on genetic interrelationships, origin and migratory routes of this medically important emerging tapeworm. [ABSTRACT FROM AUTHOR]

The microsatellite markers were designed for the giant liver fluke, Fascioloides magna, veterinary important liver parasite of free-living and domestic ruminants. Due to its geographic distribution (five enzootic regions across USA and Canada, three permanent European foci) and invasive character, F. magna is an interesting model for population genetics. Out of 667 amplicon candidates generated after NGS, 118 provided the best resolution and were tested with PCR analysis. In total, 56 yielded PCR products of expected size and in 36 of them the declared repetitive motif was identified by Sanger sequencing. After fragment analysis, 12 loci were proved to be polymorphic in individuals from one tested European and four North American populations. These loci were selected for setup of multiplex STR assays and utilized in genotyping of larger sample cohort. The outputs of statistical analyses indicate further global application of 11 conclusive loci in population genetics of the parasite [ABSTRACT FROM AUTHOR]