In the present work, a morphological and biometrical study of whipworms Trichuris Roederer, 1761 (Nematoda: Trichuridae) parasitizing Colobus guereza kikuyensis has been carried out. Biometrical and statistical data showed that the mean values of individual variables between Trichuris suis and Trichuris sp. from C. g. kikuyensis differed significantly (P < 0.001) when Student's t test was performed: seven male variables (width of esophageal region of body, maximum width of posterior region of body, width in the place of junction of esophagus and the intestine, length of bacillary stripes, length of spicule, length of ejaculatory duct, and distance between posterior part of testis and tail end of body) and three female variables (width of posterior region of body, length of bacillary stripes, and distance of tail end of body and posterior fold of seminal receptacle). The combination of these characters permitted the discrimination of T. suis with respect to Trichuris sp. from C. g. kikuyensis, suggesting a new species of Trichuris. Furthermore, males of Trichuris sp. from C. g. kikuyensis showed a typical subterminal pericloacal papillae associated to a cluster of small papillae that were absent in males of T. suis, while females of Trichuris from Colobus appeared with a vulval region elevated/over-mounted showing a crater-like appearance. The everted vagina showed typical triangular sharp spines by optical microscopy and SEM. Thus, the existence of a new species of Trichuris parasitizing C. g. kikuyensis has been proposed.
A molecular phylogenetic hypothesis is presented for the genus Trichuris based on sequence data from the mitochondrial cytochrome c oxidase 1 (cox1) and ribosomal 18S genes. The taxa consisted of different described species and several host-associated isolates (undescribed taxa) of Trichuris collected from hosts from Spain. Sequence data from mitochondrial cox1 (partial gene) and nuclear 18S near-complete gene were analyzed by maximum likelihood and Bayesian inference methods, as separate and combined datasets, to evaluate phylogenetic relationships among taxa. Phylogenetic results based on 18S ribosomal DNA (rDNA) were robust for relationships among species; cox1 sequences delimited species and revealed phylogeographic variation, but most relationships among Trichuris species were poorly resolved by mitochondrial sequences. The phylogenetic hypotheses for both genes strongly supported monophyly of Trichuris, and distinct genetic lineages corresponding to described species or nematodes associated with certain hosts were recognized based on cox1 sequences. Phylogenetic reconstructions based on concatenated sequences of the two loci, cox1 (mitochondrial DNA (mtDNA)) and 18S rDNA, were congruent with the overall topology inferred from 18S and previously published results based on internal transcribed spacer sequences. Our results demonstrate that the 18S rDNA and cox1 mtDNA genes provide resolution at different levels, but together resolve relationships among geographic populations and species in the genus Trichuris.
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.
In the present work, a comparative morphological, biometrical and molecular study of Ctenocephalides spp. isolated from dogs (Canis lupus familiaris) from different geographical regions (Spain, Iran, and South Africa) has been carried out. The internal transcribed spacer 1 (ITS1) sequences of Ctenocephalides felis and Ctenocephalides canis collected from dogs from different geographical regions have been determined to clarify the taxonomic status of these species and to assess intraspecific variation and interspecific sequence differences. In addition, a phylogenetic analysis based on ITS1 sequences has been performed. Four different morphological populations were observed in the individuals of C. felis collected from dogs from different geographical locations. Nevertheless, the comparative study of the ITS1 sequences of the different morphological populations observed in C. felis did not show molecular differences. The results showed clear molecular differences between C. felis and C. canis and some specific recognition sites for endonucleases were detected between both species. Thus, BfrBI and DraI sites have diagnostic value for specific determination in C. felis. The phylogenetic tree based on the ITS1 sequences of C. felis and C. canis revealed that all the populations of C. felis from different geographical regions clustered together and separated, with high bootstrap values, from C. canis. We conclude that ITS1 region is a useful tool to approach different taxonomic and phylogenetic questions in Ctenocephalides species.
Canine demodicosis is a severe and highly prevalent dermatologic disease in dogs. Pet dogs can be affected by three recognized Demodex species that can produce clinical effects. In this paper, three morphological types of Demodex mites have been isolated from Spanish dogs. A complete morphobiometrical study of each one has been carried out. Morphological and biometrical studies revealed three closely related populations with some distinctive characteristics and could be identified as Demodex canis, Demodex injai, and Demodex sp. "cornei." Furthermore, one population of D. canis from China, different populations of Demodex folliculorum from human skin (Spain and China), D. folliculorum from human eyelashes (Spain), and Demodex brevis from human skin (China) were considered to find out the level of variation between different species and geographical origin. The aim of the present study is to assess the usefulness of mitochondrial DNA molecular markers in establishing phylogenetic relationships and resolve taxonomic questions in Demodex mites. Molecular studies based on the amplification and sequencing of the 16S rDNA and cytochrome oxidase I mitochondrial genes did not show clear differences between the three morphotypes considered. Furthermore, phylogenetic relationships in Demodex mites were analyzed. The resulting phylogenetic trees show that Demodex species from dogs were gathered together, and populations of D. folliculorum from humans appear together in a different branch; however, D. brevis from humans seemed to be more distant. Our results show that cytochrome oxidase I region is a useful tool to solve different taxonomic questions at the species and population level and to infer phylogenetic relationships in Demodex species. However, 16S mitochondrial rDNA seems a good marker for comparisons at an interspecies level, but not at a population level in this group of mites. Furthermore, from genetic distance and divergence data, we would suggest that D. canis, D. injai, and Demodex sp. cornei are polymorphisms of the same species.
Populations of Trichuris spp. isolated from six species of sigmodontine rodents from Argentina were analyzed based on morphological characteristics and ITS2 (rDNA) region sequences. Molecular data provided an opportunity to discuss the phylogenetic relationships among the Trichuris spp. from Noth and South America (mainly from Argentina). Trichuris specimens were identified morphologically as Trichuris pardinasi, T. navonae, Trichuris sp. and Trichuris new species, described in this paper. Sequences analyzed by Maximum Parsimony, Maximum Likelihood and Bayesian inference methods showed four main clades corresponding with the four different species regardless of geographical origin and host species. These four species from sigmodontine rodents clustered together and separated from Trichuris species isolated from murine and arvicoline rodents (outgroup). Different genetic lineages observed among Trichuris species from sigmodontine rodents which supported the proposal of a new species. Moreover, host distribution showed correspondence with the different tribes within the subfamily Sigmodontinae.
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