title: Development and characterization of multiplex panels of polymorphic microsatellite loci in giant liver fluke Fascioloides magna (Trematoda: Fasciolidae), using next-generation sequencing approach.
abstract: 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.
authors: Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, Mlynská dolina B-2, 84215 Bratislava, Slovakia; Institute of Molecular Biomedicine, Faculty of Medicine, Comenius University, Sasinkova 4, 81108 Bratislava, Slovakia; Geneton Ltd., Ilkovičova 3, 84104 Bratislava, Slovakia.
authors: Gabriel
authors: G
authors: Minárik
authors: Institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 04001 Košice, Slovakia.
authors: Eva
authors: E
authors: Bazsalovicsová
authors: Institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 04001 Košice, Slovakia.
authors: Ludmila
authors: L
authors: Zvijáková
authors: Biology Centre ASCR, Institute of Parasitology, Faculty of Science, University of South Bohemia, Branišovská 31, 37005 České Budějovice, Czech Republic.
authors: Jan
authors: J
authors: Stefka
authors: Institute of Molecular Biomedicine, Faculty of Medicine, Comenius University, Sasinkova 4, 81108 Bratislava, Slovakia.
authors: Lenka
authors: L
authors: Pálková
authors: Institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 04001 Košice, Slovakia. Electronic address: hromadova@saske.sk.
authors: Ivica
authors: I
authors: Králová-Hromadová
mesh_terms: D000818:Animals; D005212:Fasciolidae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D018895:Microsatellite Repeats; D011110:Polymorphism, Genetic; D012418:Ruminants / Q000469:parasitology*; D014201:Trematode Infections / Q000469:parasitology / Q000662:veterinary*
keywords: Fascioloides magna; Invasive parasite; Microsatellites; Population genetics; Short tandem repeats