The complete internal transcribed spacer 1 (ITS1), 5.8S rDNA and ITS2 region of the ribosomal DNA from 11 species of rhinonyssid mites ( Tinaminyssus columbae, T. minisetosum, T. sartbaevi, T. bubulci, T. melloi, T. streptopelioides, Sternostoma fulicae, S. boydi, S. strandtmanni, S. turdi, Rhinonyssus tringae) were sequenced to assess the utility of this genomic region in resolving taxonomic questions in this group and to estimate phylogenetic relationships between species. Two different geographic locations of T. melloi and T. streptopelioides were analyzed to detect intraspecies variation. Our study shows that ribosomal sequences can help to discriminate between T. melloi and T. sartbaevi, which are morphologically very close and difficult to separate by classic methods. The resulting phylogenetic tree shows some differences from the current taxonomy of the family Rhinonyssidae. This study appeals for the revision of the taxonomic status of S. boydi and closely related species which parasitize aquatic birds and suggests the synonymy of S. boydi and S. strandtmanni, despite the different hosts of the two mites.
A phylogeographic study was carried out of Trichuris muris, nematode parasitizing Murinae rodents from the Muridae family, isolated from four different hosts and from different geographical regions of Europe by amplification and sequencing of the ITS1-5.8S-ITS2 fragment of the ribosomal DNA. T. muris was found in the Apodemus sylvaticus, Apodemus flavicollis, Mus domesticus, and Rattus rattus rodents. The molecular results confirm the presence of DNA polymorphisms among T. muris isolates from Europe. The present study shows two clear-cut geographical and genetic lineages: one of them is widespread from northern Spain (Catalonia) to Denmark (Western European region), while the second is widespread in the Eastern European region (Croatia, Rumania, and Turkey). These two genotypes can be easily distinguished by a PCR-RFLP analysis of this sequence with the ApalI restriction enzyme. Moreover, networks and phylogenetic reconstructions also reveal that T. muris from various Murinae rodents did not differentiate according to the host species that they parasitize. Furthermore, T. muris isolated from The Canary Islands revealed a typical haplotype (H6) only present in The Canary Islands and not in continental Europe. It is suggested that one haplotype from La Gomera Island is the ancestor of T. muris in the Canary Islands.
Trichuris muris has been isolated from murid hosts ( Apodemus sylvaticus and Mus musculus) and Trichuris arvicolae from arvicolid rodents in Barcelona, Spain. Genomic DNA was isolated and the ITS1-5.8S-ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced using polymerase chain reaction techniques. The ITS2 of both populations isolated from Apodemus and Mus was 382 nucleotides in length and had a GC content of about 60.73%, while the ITS2 of T. arvicolae was 442 nucleotides in length and had a GC content of about 59.8%. Furthermore, the ITS1 of Trichuris from murids was 448 nucleotides in length and had a GC content of about 56.47%, while T. arvicolae was 446 nucleotides in length and had 57.62% of GC content. A total of 161 and 173 nucleotides were observed along the 5.8S gene of T. murisand T. arvicolae, respectively; This difference in nucleotides was due to the insertion of a DNA segment (transposon) in the 5.8S sequence of the latter species. Slight intraindividual and intraspecific variations were detected in the rDNA of both species. The presence of microsatellites was observed in all of the individuals assayed. Sequence analysis of the internal transcribed spacers and the 5.8S gene demonstrated no sequence differences between T. muris isolated from both of its murid hosts. Nevertheless, clear differences were detected between the ITS2, ITS1 and 5.8S gene of T. muris and T. arvicolae. This corroborates the existence of two separate Trichuris species in murid and arvicolid hosts. Furthermore, a phylogenetic analysis was carried out and endonucleases restriction maps were elaborated for both species.
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.
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