Molecular identification of Trichuris vulpis and Trichuris suis isolated from different hosts.

Trichuris suis was isolated from the cecum of two different hosts (Sus scrofa domestica -- swine and Sus scrofa scrofa -- wild boar) and Trichuris vulpis from dogs in Sevilla, Spain. Genomic DNA was isolated and internal transcribed spacers (ITS)1-5.8S-ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced using polymerase chain reaction techniques. The sequence of T. suis from both hosts was 1,396 bp in length while that of T. vulpis was 1,044 bp. ITS1 of both populations isolated of T. suis was 661 nucleotides in length, while the ITS2 was 534 nucleotides in length. Furthermore, the ITS1 of T. vulpis was 410 nucleotides in length, while the ITS2 was 433 nucleotides in length. One hundred fifty-four nucleotides were observed along the 5.8S gene of T. suis and T. vulpis. Intraindividual and intraspecific variations were detected in the rDNA of both species. The presence of microsatellites was observed in all the individuals assayed. Sequence analysis of the ITSs and the 5.8S gene has demonstrated no sequence differences between T. suis isolated from both hosts (S. scrofa domestica -- swine and S. scrofa scrofa -- wild boar). Nevertheless, clear differences were detected between the ITS1 and ITS2 of T. suis and T. vulpis. Furthermore, a comparative molecular analysis between both species and the previously published ITS1-5.8S-ITS2 sequence data of Trichuris ovis, Trichuris leporis, Trichuris muris, Trichuris arvicolae, and Trichuris skrjabini was carried out. A common homology zone was detected in the ITS1 sequence of all species of trichurids.

Utility of ITS1-5.8S-ITS2 and 16S mitochondrial DNA sequences for species identification and phylogenetic inference within the Rhinonyssus coniventris species complex (Acari: Rhinonyssidae).

The complete internal transcribed spacer 1 (ITS1), 5.8S rDNA, and ITS2 region of the ribosomal DNA and a 390-bp region of the 16S rDNA gene from five taxa belonging to Rhinonyssus (Rhinonyssus vanellus, Rhinonyssus tringae, Rhinonyssus neglectus, Rhinonyssus echinipes from Kentish plover, and Rhinonyssus echinipes from grey plover) were sequenced to examine the level of sequence variation and the taxonomic levels to show utility in phylogeny estimation. Our data show that these molecular markers can help to discriminate between species and populations included in the Rhinonyssus coniventris complex (R. tringae, R. neglectus, R. echinipes), which are morphologically very close and difficult to separate by classic methods. A comparative study with sequences from other rhinonyssid mites previously published was also carried out. The resulting phylogenetic tree inferred from ITS1-5.8S-ITS2 region sequences obtained in this paper, together with those from other 11 taxa of rhinonyssid, shows slight differences from the current taxonomy of the Rhinonyssidae. This study appeals for the revision of the taxonomic status of the R. coniventris complex, as well as for the species included within it.