Fasciola hepatica: identification of molecular markers for resistant and susceptible Pseudosuccinea columella snail hosts

Abstract

Protein electrophoresis, RAPD-PCR and nuclear rDNA ITS sequencing were performed to search for genetic differences between Pseudosuccinea columella snails susceptible and resistant to Fasciola hepatica infection. Of the 21 enzymatic loci analyzed in both populations, none of them exhibited neither within- or between-group variation. Such an absence of enzyme polymorphism support the hypothesis of selfing as the “prevalent” mating system for this hermaphroditic species. Conversely, the RAPD profiles displayed clear differences between susceptible and resistant isolates for 17 of the 26 primers tested while no within-group variation was detected. rDNA ITS sequence analysis from snails of each isolates showed only two bases that differed between groups accounting for a 0.17% of variation confirming that suceptible and resistant snails belong to the same species. This is the first time that a genetic variation using RAPD markers is demonstrated between susceptible and resistant lymnaeid snails vis-a-vis of F. hepatica infection in absence of experimental selection. Index Descriptors and Abbreviations: DNA, desoxyribonucleic acid; rDNA, ribosomal DNA; ITS, internal transcribed spacer; PCR, polymerase chain reaction; RAPD, random amplified polymorphic DNA; S, similitude coefficient