Study by Ribosomal DNA Its 2 Sequencing and RAPD Analysis on the Systematics of Four Metastrongylus Species (Nematoda: Metastrongyloidea)

Four species of lungworms of the Metastrongylus genus are found sympatrically in the lungs of the wild boars (Sus scrofa L.) on the Chambord game reserve (France): Metastrongylus asymmetricus, M. confusus, M. pudendotectus, and M. salmi. These species are difficult to identify using morphological characters alone, and epidemiological evidence raises doubt about the existence of 4 distinct species. Two molecular approaches (sequencing of ribosomal DNA Internal Transcribed Spacer 2 [ITS 2] and Random Amplified Polymorphic DNA assay [RAPD]) were used to evaluate the genetic similarities between the 4 taxa. The ITS 2 sequences of M. salmi and M. confusus were identical, whereas the sequences of the other species were distinct. On the other hand, RAPD analysis indicated unambiguously that the 4 species are genetically different. These results reveal the limitation of using only 1 molecular approach for taxonomic studies and indicate the need to examine different regions (ribosomal, mitochondrial DNA) in the genome of a species in complement with morphological and epidemiological data.