Most cited article - PubMed ID 24959976
Development and characterization of multiplex panels of polymorphic microsatellite loci in giant liver fluke Fascioloides magna (Trematoda: Fasciolidae), using next-generation sequencing approach
BACKGROUND: Monogenean flatworms are the main ectoparasites of fishes. Representatives of the species-rich families Gyrodactylidae and Dactylogyridae, especially those infecting cichlid fishes and clariid catfishes, are important parasites in African aquaculture, even more so due to the massive anthropogenic translocation of their hosts worldwide. Several questions on their evolution, such as the phylogenetic position of Macrogyrodactylus and the highly speciose Gyrodactylus, remain unresolved with available molecular markers. Also, diagnostics and population-level research would benefit from the development of higher-resolution genetic markers. We aim to offer genetic resources for work on African monogeneans by providing mitogenomic data of four species (two belonging to Gyrodactylidae, two to Dactylogyridae), and analysing their gene sequences and gene order from a phylogenetic perspective. RESULTS: Using Illumina technology, the first four mitochondrial genomes of African monogeneans were assembled and annotated for the cichlid parasites Gyrodactylus nyanzae, Cichlidogyrus halli, Cichlidogyrus mbirizei (near-complete mitogenome) and the catfish parasite Macrogyrodactylus karibae (near-complete mitogenome). Complete nuclear ribosomal operons were also retrieved, as molecular vouchers. The start codon TTG is new for Gyrodactylus and for Dactylogyridae, as is the incomplete stop codon TA for Dactylogyridae. Especially the nad2 gene is promising for primer development. Gene order was identical for protein-coding genes and differed between the African representatives of these families only in a tRNA gene transposition. A mitochondrial phylogeny based on an alignment of nearly 12,500 bp including 12 protein-coding and two ribosomal RNA genes confirms that the Neotropical oviparous Aglaiogyrodactylus forficulatus takes a sister group position with respect to the other gyrodactylids, instead of the supposedly 'primitive' African Macrogyrodactylus. Inclusion of the African Gyrodactylus nyanzae confirms the paraphyly of Gyrodactylus. The position of the African dactylogyrid Cichlidogyrus is unresolved, although gene order suggests it is closely related to marine ancyrocephalines. CONCLUSIONS: The amount of mitogenomic data available for gyrodactylids and dactylogyrids is increased by roughly one-third. Our study underscores the potential of mitochondrial genes and gene order in flatworm phylogenetics, and of next-generation sequencing for marker development for these non-model helminths for which few primers are available.
- Keywords
- Cichlidae, Cichlidogyrus, Clariidae, Gene order, Gyrodactylus, Macrogyrodactylus, Mitogenome, Monogenea, Monopisthocotylea, Phylogenomics,
- MeSH
- Cichlids parasitology MeSH
- Phylogeny MeSH
- Genome, Mitochondrial MeSH
- Mitochondria classification genetics MeSH
- Platyhelminths genetics MeSH
- Gene Order MeSH
- DNA, Protozoan chemistry isolation & purification metabolism MeSH
- Protozoan Proteins classification genetics MeSH
- RNA, Ribosomal classification genetics MeSH
- Sequence Analysis, DNA MeSH
- High-Throughput Nucleotide Sequencing MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- DNA, Protozoan MeSH
- Protozoan Proteins MeSH
- RNA, Ribosomal MeSH
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.
- Keywords
- Fascioloides magna *, Genetic interrelationships *, Giant liver fluke *, Microsatellites *, Migratory routes *, Parasite *,
- MeSH
- Global Health MeSH
- Fasciolidae classification genetics isolation & purification MeSH
- Genotyping Techniques MeSH
- Trematode Infections epidemiology transmission MeSH
- Microsatellite Repeats MeSH
- Deer * MeSH
- Zoonoses epidemiology parasitology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Europe epidemiology MeSH
- North America epidemiology MeSH
BACKGROUND: Population structure and genetic interrelationships of giant liver fluke Fascioloides magna from all enzootic North American regions were revealed in close relation with geographical distribution of its obligate definitive cervid hosts for the first time. METHODS: Variable fragments of the mitochondrial cytochrome c oxidase subunit I (cox1; 384 bp) and nicotinamide dehydrogenase subunit I (nad1; 405 bp) were applied as a tool. The concatenated data set of both cox1 and nad1 sequences (789 bp) contained 222 sequences that resulted in 50 haplotypes. Genetic data were analysed using Bayesian Inference (BI), Maximum Likelihood (ML) and Analysis of Molecular Variance (AMOVA). RESULTS: Phylogenetic analysis revealed two major clades of F. magna, which separated the parasite into western and eastern populations. Western populations included samples from Rocky Mountain trench (Alberta) and northern Pacific coast (British Columbia and Oregon), whereas, the eastern populations were represented by individuals from the Great Lakes region (Minnesota), Gulf coast, lower Mississippi, and southern Atlantic seaboard region (Mississippi, Louisiana, South Carolina, Georgia, Florida) and northern Quebec and Labrador. Haplotype network and results of AMOVA analysis confirmed explicit genetic separation of western and eastern populations of the parasite that suggests long term historical isolation of F. magna populations. CONCLUSION: The genetic makeup of the parasite's populations correlates with data on historical distribution of its hosts. Based on the mitochondrial data there are no signs of host specificity of F. magna adults towards any definitive host species; the detected haplotypes of giant liver fluke are shared amongst several host species in adjacent populations.
- MeSH
- Fasciola hepatica classification enzymology genetics isolation & purification MeSH
- Fascioliasis epidemiology parasitology veterinary MeSH
- Phylogeny MeSH
- Genetic Variation MeSH
- Molecular Sequence Data MeSH
- Ruminants parasitology MeSH
- Helminth Proteins genetics metabolism MeSH
- Electron Transport Complex IV genetics metabolism MeSH
- Deer MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Quebec MeSH
- United States MeSH
- Names of Substances
- Helminth Proteins MeSH
- Electron Transport Complex IV MeSH