Identification of Chabertiidae (Nematoda, Strongylida) by PCR-RFLP based method: A new diagnostic tool for cross transmission investigation between domestic and wild ruminants in France

We describe a PCR-RFLP-based method that allows reliable identification of four species of nematode parasites presenting similar infective third-stage larvae (L3) with a flagelliform tail and more than 16 intestinal cells, commonly observed in gastrointestinal tract of ruminants in France. Molecular analysis of the second internal transcribed spacer (ITS2) of ribosomal DNA, considered as a specific marker for Strongylida, revealed four robust monophyletic clades corresponding to species Chabertia ovina, Oesophagostomum sikae, Oesophagostomum radiatum and Oesophagostomum venulosum. One restriction enzyme (DdeI) was used to digest this domain, and we observed four different and clear digestion patterns according to these species (adults or larvae). Hence, this new method is a good tool easy to use for veterinary laboratories to characterize the different species, and allows considering possible cross transmission between domestic and wild ruminants, especially cervids often incriminated as potential reservoir of parasites for cattle. Moreover, thanks to this new tool, necroscopic analyses could be substituted by coprological methods, a non-invasive approach. (C) 2014 Elsevier B.V. All rights reserved.