The use of mitochondrial and nuclear sequences in prospecting for cryptic species in Tabascotrema verai (Digenea: Cryptogonimidae), a parasite of Petenia splendida (Cichlidae) in Middle America.

In this study, we used sequences of mitochondrial and nuclear markers to test the hypothesis that Tabascotrema verai Lamothe-Argumedo and Pineda-López, 1990, a parasite of the cichlid Petenia splendida Günther, 1862 in Middle America, represents a single species, following a molecular prospecting approach. One-hundred and five individuals of T. verai were collected from the intestine of 43 specimens of P. splendida in eight localities of Mexico, Belize and Guatemala. A portion (n=53) of the collected specimens was used for molecular studies, whereas the remaining individuals were processed for morphological studies. Fifty-three partial sequences of the cytochrome c oxidase subunit I (cox1) gene were obtained. In addition, 36 sequences of the ITS (internal transcribed spacers 1, 5.8S rRNA gene, and ITS2) and of the 28S rRNA gene were generated from most of the specimens that showed nucleotide variation with the cox1 gene. A haplotype network obtained from cox1 sequences revealed three independent groups (haplogroups 1, 2, and 3). Independent phylogenetic analyses performed with maximum likelihood and Bayesian inference of cox1, ITS and the 28S rRNA gene recovered three genetically-distinct and reciprocally-monophyletic groups, corresponding with the 3 haplogroups obtained in the haplotype network. Values of genetic divergence between clades for cox1 sequences ranged between 8.3% and 11.9%, while for ITS and the 28S rRNA gene, these ranged from 0.08% to 1.2% and from 0.3% to 0.4%, respectively. Morphological observations, and measurements of 23 characters of 44 mounted individuals, showed that no morphological differences exist between individuals from the eight collecting sites, and that the ranges of most morphological traits overlap. Our results suggest that the digenean T. verai represents a complex of cryptic species; the haplotype network, phylogenetic analyses, and genetic differences, along with the morphological stasis recorded here support this notion. Finally, the three recovered lineages showed neither geographical association nor correlation with drainage basins.

The systematic position of Wallinia spp. and Margotrema spp. (Digenea), parasites of Middle-American and Neotropical freshwater fishes, based on the 28S ribosomal RNA gene.

The systematic relationships of aspinose allocreadiid-like digeneans from Middle-American and Neotropical freshwater fishes, such as Wallinia Pearse, 1920, Margotrema Lamothe-Argumedo, 1970, Magnivitellinum Kloss, 1966 and other related genera, remain enigmatic. Wallinia (2 spp.) and Margotrema (2 spp.) have been placed in the subfamily Walliniinae, within the Macroderoididae. Recent descriptions of species in these genera have followed this classification but have raised questions about their affinities with macroderoidids. Partial sequences of the 28S rRNA gene of Wallinia chavarriae Choudhury, Hartvigsen & Brooks, 2002 and Margotrema bravoae Lamothe Argumedo, 1970, as representatives of their genera, were used in a phylogenetic analysis along with the available sequences of macroderoidids and allocreadiids. Results from Maximum Parsimony, Maximum Likelihood and Bayesian Analyses were identical in placing both genera into one clade along with allocreadiids rather than the macroderoidids. Margotrema was the sister group of Crepidostomum Braun, 1900, and Wallinia was the sister group of both Margotrema and Crepidostomum combined. Bootstrap and Bremer values, as well as posterior probabilities derived from Bayesian analysis strongly supported these relationships in three analyses.