Characterisation of a novel panel of polymorphic microsatellite loci for the liver fluke, Fasciola hepatica, using a next generation sequencing approach

Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases
Krystyna CwiklinskiJane E Hodgkinson

Abstract

The liver fluke, Fasciola hepatica is an economically important pathogen of sheep and cattle and has been described by the WHO as a re-emerging zoonosis. Control is heavily reliant on the use of drugs, particularly triclabendazole and as a result resistance has now emerged. The population structure of F. hepatica is not well known, yet it can impact on host-parasite interactions and parasite control with drugs, particularly regarding the spread of triclabendazole resistance. We have identified 2448 potential microsatellites from 83 Mb of F. hepatica genome sequence using msatfinder. Thirty-five loci were developed and optimised for microsatellite PCR, resulting in a panel of 15 polymorphic loci, with a range of three to 15 alleles. This panel was validated on genomic DNA from 46 adult F. hepatica; 38 liver flukes sourced from a Northwest abattoir, UK and 8 liver flukes from an established isolate (Shrewsbury; Ridgeway Research). Evidence for null alleles was found at four loci (Fh_1, Fh_8, Fh_13 and Fh_14), which showed markedly higher levels of homozygosity than the remaining 11 loci. Of the 38 liver flukes isolated from cattle livers (n=10) at the abattoir, 37 genotypes were identified. Using a multiplex approach all 15 loci ...Continue Reading

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Citations

Jan 15, 2016·Journal of Helminthology·A A VázquezS Hurtrez-Boussès
Jun 18, 2016·Parasite Immunology·K CwiklinskiJ P Dalton
Mar 21, 2017·Trends in Parasitology·Janneke Wit, John S Gilleard
Jun 28, 2019·Grass and Forage Science : the Journal of the British Grassland Society·Bethan C JohnJane E Hodgkinson
Feb 14, 2021·International Journal for Parasitology·Nicola J BeesleyJane E Hodgkinson

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Datasets Mentioned

BETA
PRJEB8349

Methods Mentioned

BETA
PCR
electrophoresis
genotyping
PCRs

Software Mentioned

BLAST
Peak Scanner
GENEPOP
msatfinder
Genetix
Micro
Checker

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