Exploring the genetic structure of Parastrigea diovadena Dubois and Macko, 1972 (Digenea: Strigeidae), an endoparasite of the white ibis, Eudocimus albus, from the Neotropical region of Mexico.

Parastrigea diovadena Dubois and Macko, 1972, is an allogenic trematode species that infects the intestine of white ibis. This widely distributed Neotropical species has been studied poorly, and nothing is known about its population genetic structure. In the current study, we attempt to fill this gap for the first time and to explore the genetic diversity in P. diovadena populations from three biogeographic provinces (Sierra Madre Oriental, Sierra Madre Occidental, and Sierra Madre del Sur) in the Neotropical region of Mexico. Newly generated sequences of the internal transcribed spacers (ITS) from ribosomal DNA and cytochrome c oxidase subunit 1 (cox 1) from mitochondrial DNA were compared with sequences available from the GenBank data set. Phylogenetic analyses performed with the ITS and cox 1 data sets using maximum likelihood and Bayesian inference unequivocally showed that new sequences of P. diovadena recovered from the white ibis formed a clade with other sequences of specimens previously identified as P. diovadena. The intraspecific genetic divergence among the isolates was very low, ranging from 0 to 0.38% for ITS and from 0 to 1.5% for cox 1, and in combination with the phylogenetic trees confirmed that the isolates belonged to the same species. The cox 1 haplotype network (star-shaped) inferred with 62 sequences revealed 36 haplotypes. The most frequent haplotype (H3, n = 18) corresponded to specimens from all the populations (except Tecolutla, Veracruz). In addition to the common haplotype, we identified four other shared haplotypes (H2, H9, H12, and H14) and 31 unique haplotypes (singlets). In addition, high haplotype diversity (Hd = 0.913), low nucleotide diversity (Pi = 0.0057), and null genetic differentiation or population structure (Fst = 0.0167) were found among the populations from the three biogeographic provinces. The results suggest that the biology of the definitive host has played a key role in the population genetic structure of Parastrigea diovadena in the Neotropical region of Mexico.

Taxonomic status of Rhabdochona ictaluri (Nematoda: Rhabdochonidae) based on molecular and morphological evidence.

The genus Rhabdochona includes more than 100 species infecting freshwater fishes in all zoogeographical regions of the world. In Mexico, 12 nominal species of Rhabdochona have been recorded. Of these, Rhabdochona ictaluri was originally described as a parasite of endemic catfishes of the family Ictaluridae; however, the species was later considered on morphological grounds as a junior synonym of Rhabdochona kidderi. In this study, newly sampled specimens of R. ictaluri were obtained from the type host and type locality and were used to perform a detailed morphological analysis and molecular phylogenetic inferences through one mitochondrial and two nuclear genes; data were used in an integrative taxonomy context to test the taxonomic status of R. ictaluri. This approach proved to be very useful to confirm the validity of this species, and robust species limits were established between these two putative species considering morphology, molecular data, host association, and biogeography.

Description of two new species of allocreadiid trematodes (Digenea: Allocreadiidae) in middle American freshwater fishes using an integrative taxonomy approach.

Integrative taxonomy uses several sources of information to establish more robust species delimitation criteria. In this study, we followed that approach to describe two new species of allocreadiid trematodes from freshwater fish distributed in locations across Middle America. The new species were first recognized by using sequences of the domains D1-D3 of the ribosomal 28S rRNA gene and then, morphological data (light and scanning electron microscopy (SEM)), host association, and geographical distribution were considered as additional sources of information to complement the species delimitation and description. Auriculostoma tica n. sp. was found in the intestine of Gymnotus maculosus Albert and Miller in Costa Rica, while Wallinia anindoi n. sp. was found in the intestine of Astyanax aeneus (Günther) in Oaxaca and Chiapas, Mexico, as well as in a location of Guatemala. A phylogenetic analysis combining newly generated sequences of the two new species along with those available for other allocreadiids in GenBank, revealed that isolates of each species are reciprocally monophyletic, and also their interrelationships: Au. tica n. sp. is a sister taxon of Auriculostoma totonacapanensis Razo-Mendivil et al., 2014, and W. anindoi n. sp. is a sister taxon of Wallinia brasiliensis Dias, et al., 2018. Genetic distances for the 28S rRNA gene were estimated among the American species of allocreadiids and further supported the validity of the new species.

Filling the gaps in the classification of the Digenea Carus, 1863: systematic position of the Proterodiplostomidae Dubois, 1936 within the superfamily Diplostomoidea Poirier, 1886, inferred from nuclear and mitochondrial DNA sequences.

The Diplostomida Olson, Cribb, Tkach, Bray & Littlewood, 2003 is the less diverse order of the two orders within the subclass Digenea Carus, 1863 and is currently classified into three superfamilies, i.e. Brachylaimoidea Joyeux & Foley, 1930, Diplostomoidea Poirier, 1886, and Schistosomatoidea Stiles & Hassall, 1898. Although the suprageneric-level relationships have been elucidated with the use of molecular markers, the lack of representation of some groups obscure the phylogenetic relationships among families, rendering the classification unstable. Here, we tested the phylogenetic position of the family Proterodiplostomidae Dubois, 1936 based on partial 28S rDNA and complete 18S rDNA sequences for Crocodilicola pseudostoma (Willemoes-Suhm, 1870), a crocodile parasite that has been found as a progenetic metacercaria parasitising the pale catfish Rhamdia guatemalensis (Günther) in Mexico and in other siluruforms in the Neotropics. We augmented the representation of the species, genera and families within the Diplostomida, including mostly representatives of the superfamily Diplostomoidea, and assembled a dataset that contains 49 species for the 28S rRNA gene, and 45 species for the 18S rRNA gene. Additionally, we explored the phylogenetic signal of the mitochondrial gene cox1 in reconstructing the phylogenetic relationships of selected members of the superfamily. Our analyses showed that the family Proterodiplostomidae is the sister taxon to the paraphyletic Diplostomidae Poirier, 1886 and Strigeidae Railliet, 1919, with Cyathocotylidae Mühling, 1898 + Brauninidae Wolf, 1903 as their sister group. Analysis of concatenated 18S + 28S sequences revealed the Liolopidae Odhner, 1912 as the basal group of the superfamily Diplostomoidea, although analyses of independent datasets showed that the position of this family remains uncertain. Analysis based on cox1 unequivocally resolved the Proterodiplostomidae as the sister taxon to the Diplostomidae and Strigeidae, although the Cyathocotylidae was nested in a different clade, along with brachylaimoids and schistosomatoids.

Phylogenetic position of Magnivitellinum Kloss, 1966 and Perezitrema Barus & Moravec, 1967 (Trematoda: Plagiorchioidea: Macroderoididae) inferred from partial 28S rDNA sequences, with the establishment of Alloglossidiidae n. fam.

The systematic position of two genera of Macroderoididae McMullen, 1937, Perezitrema Barus & Moravec, 1967 and Magnivitellinum Kloss, 1966 is reviewed based on a phylogenetic analysis of the interrelationships of 15 species of the family allocated into six genera, along with 44 species of plagiorchioid trematodes, using partial sequences of the 28S rRNA gene. Sequences were analysed through parsimony, maximum likelihood and Bayesian inference. The obtained topologies show Perezitrema as the sister taxon of three species of Macroderoides Pearse, 1924; the latter genus appears to be paraphyletic since another three species are not included in this group. Instead, Magnivitellinum was placed as the sister taxon of Alloglossidium Simer, 1929. These relationships are well supported by high bootstrap and posterior probability values. The resulting trees demonstrate that the family Macroderoididae, as currently conceived in taxonomic treatments, is not monophyletic. Magnivitellinum simplex Kloss, 1966 and Alloglossidium spp. were nested as sister taxa of members of the family Leptophallidae Dayal, 1938, whereas Perezitrema bychowskii Barus & Moravec, 1967 and species of Macroderoides and Paramacroderoides Venard, 1941 were grouped with Auridistomum chelydrae (Stafford, 1900), a monotypic member of Auridistomidae Stunkard, 1924. Based on our results, a new family, Alloglossidiidae n. fam. was established to accommodate the genera Magnivitellinum and Alloglossidium.

Cathetocephalus badilloi n. sp. (Cestoda: Cathetocephalidea) from the spinner shark Carcharhinus brevipinna off the coast of Yucatan, Mexico.

This study describes a new species belonging to the genus Cathetocephalus from the spinner shark Carcharhinus brevipinna off the coast of Yucatan, Mexico. Cathetocephalus badilloi n. sp. has a soft scolex transversely positioned to the long axis of the strobila. The scolex is composed of an apical pad, papillate band and rugose base. The papillate band has two papillary segments; the upper segment has numerous papillae, positioned one next to the other in an irregular and heterogeneous way with a spongy texture that remains along the band. The lower papillary segment has numerous papillae positioned one next to the other, without space between them. This papillary band presents flattened rectangular papillae with a division located at the top of each the papilla, giving it the appearance of a molar tooth. Using the 28S rDNA gene and the method of Maximum Likelihood (ML) for phylogenetic analysis, we identified the present material as a new species. We did not collect specimens with mature or gravid proglottids, but since the specific identification of the members of this genus is based on the characteristics of the scolex, our proposal of the new species is determined by the morphology of this organ as well as the molecular results obtained.

Linking phenotypic to genotypic metacestodes from Octopus maya of the Yucatan Peninsula.

Octopus maya is an endemic species of the Yucatan Peninsula. This species sustains the octopus' fishery in the region and is the only cephalopod cultured in Mexico. It is known that O. maya harbor a large richness and abundance of metacestodes that have been tentatively identified by light microscopy alone. Since the larval stages of some orders of marine cestodes lack the taxonomic characteristics shown by the adult stages and on which cestode taxonomy is based, identification down to the species level is often unattainable. Hence, the goal of this study was to characterize the parasites, for the first time, at morphological and molecular levels. A total of 60 octopuses were collected from September to December 2017 from four fishery landing ports in Yucatán: Sisal, Progreso, Dzilam de Bravo, and Rio Lagartos (15 hosts per locality). Morphology of metacestodes was characterized by light and Scanning Electron Microscopy (SEM), while the genes 18S and 28S rDNA were sequenced for molecular characterization. Based on phenotypic characters and molecular data, seven taxa of metacestodes were identified, four of them belonging of order Trypanorhyncha: Eutetrarhynchus sp., Kotorella pronosoma, Nybelinia sp., Prochristianella sp. 1; and the three remaining taxa belonging to the order Onchoproteocephalidea: Acanthobothrium sp., Phoreiobothrium sp., and Prosobothrium sp. This work provides, for the first time, molecular support to the morphological characterization of metacestodes recorded in Octopus maya.

Parasites with zoonotic potential found in commercially important fish in Tamaulipas, Northeastern Mexico.

Human population is exposed to numerous parasitic ichthyozoonoses. Although Tamaulipas state (northeastern Mexico) is well known for its fishing and aquaculture industry, there are few reports of this type of zoonosis. Therefore, it is imperative to investigate whether the parasites that affect these fish may represent a zoonotic risk for the inhabitants of the area. The objective of this study was to identify molecular and/or morphologically muscle parasites of fish from coastal locations in Tamaulipas, Mexico, and assess the risk of infection for humans. Between 2017 and 2018, 764 individual fish belonging to 28 species were examined for parasites. Collected worms were processed for their identification using morphological characteristics. In addition, partial sequences of the large subunit (28S) ribosomal RNA gene were obtained from some species to corroborate their identity. Prevalence and mean intensity of all registered infections were calculated. A total of seven species of parasites were found: cestodes (Poecilancistrium caryophyllum), trematodes (Clinostomum tataxumui, Clinostomum cichlidorum), nematodes (Eustrongylides sp., Contracaecum sp.) and pentastomids (Sebekia purdieae, Sebekia sp.). Parasites infected 10 species belonging to different fish families (Ariidae, Centrarchidae, Centropomidae, Cichlidae, Eleotridae, Ictaluridae, Mugilidae and Sciaenidae). Congeneric species of parasites or related to those registered in this study have been identified as zoonotic agents in other regions of the world. Despite the low levels of infection (2.6-16.6% prevalence and 1-5.5 parasites per infected host), there is a latent risk of transmission to humans, so it is recommended to avoid eating raw or undercooked fish meat.

Morphological and molecular identification of helminths of the greater bulldog bat Noctilio leporinus (Quiroptera: Noctilionidae) from Campeche, Mexico.

Surveys on parasites of bats from the Americas have been conducted, but information on helminths is still scarce, especially in the Neotropical region. In Mexico, there are species of bats that lack of a record for helminth species, such as members of the family Noctilionidae. The present study describes for the first time the helminths of Noctilio leporinus in Campeche, Mexico. In 2017, six specimens of N. leporinus were studied for helminths. The species identification of helminths was based on morphological studies and molecular analysis of fragments of the 28S rDNA. All bat specimens were infected for at least one helminth species. Three helminth taxa were identified: the trematode Pygidiopsis macrostomum, and the nematodes Tricholeiperia cf. proencai, and Heligmonellidae gen. sp. The morphological identification of P. macrostomum was confirmed by sequence analysis of 28S rDNA gene. The phylogeny of P. macrostomum grouped our sequence with other sequences of the same species collected in Brazil. The phylogenetic tree of Heligmonellidae gen. sp. indicated that the helminth belongs to clade formed by the species Odilia bainae, Nippostrongylus magnus and Nippostrongylus brasiliensis of the family Heligmonellidae. The phylogenetic analysis of the 28S sequences of T. cf. proencai did not show any similarity or close affinity with nematodes from which that gene has been sequenced to date. The findings of the present study increase the number of helminth species parasitizing bats in Mexico.

Phylogenetic Position of Pseudosellacotyla lutzi (Freitas, 1941) (Digenea: Cryptogonimidae), A Parasite of Hoplias malabaricus (Bloch) in South America, through 28S rDNA Sequences, and New Observations of the Ultrastructure of Their Tegument.

The freshwater fish digenean Pseudosellacotyla lutzi ( Freitas, 1941 ) Yamaguti, 1954 has had an unsettled taxonomic history, and has at various times been classified as a member of Nanophyetidae, Heterophyidae, Microphallidae, Faustulidae, and Cryptogonimidae. Nine individual specimens of the trahira, Hoplias malabaricus (Bloch, 1794), were sampled in the Paraná River basin, Paraná State, Brazil; 22 specimens of P. lutzi were collected. One specimen of P. lutzi was used to obtain a sequence of the domains D1-D3 of the 28S rRNA gene, and to perform a phylogenetic analysis to assess their position and classification within Plagiorchiida. The resulting tree unequivocally shows that the species, along with acanthostomines, belong to the Cryptogonimidae, corroborating recent findings based on the morphology of the cercariae, and in the characteristics of the life cycle. In addition, the study of the ultrastructure of the tegumental spines through scanning electron microscopy allowed us to characterize them as pectinate spines possessing 3 to 8 digitiform projections at their distal end and extending from the anterior to the posterior extremity of the body. This study also provides the first molecular data for a cryptogonimid from South America.

A new species of Auriculostoma (Trematoda: Allocreadiidae) from the intestine of Brycon guatemalensis (Characiformes: Bryconidae) from the Usumacinta River Basin, Mexico, based on morphology and 28S rDNA sequences, with a key to species of the genus.

We describe a new species of Auriculostoma Scholz, Aguirre-Macedo & Choudhury, 2004 based on several sources of information including morphology (light and scanning electron microscopy [SEM]), sequences of two nuclear genes, host association, and geographical distribution. Morphologically, the new species most closely resembles Auriculostoma astyanace Scholz, Aguirre-Macedo & Choudhury, 2004, but differs by having deeply lobated testes and cirrus-sac extending posteriorly to seminal receptacle level. Auriculostoma lobata n. sp. can be readily distinguished from all the other congeners by the combination of the following characters: testes located in tandem, testes deeply lobated, and larger body size. A phylogenetic analysis using 28S rDNA sequences along with those available for other allocreadiid trematodes, revealed that the new species is a sister taxon of A. astyanace, a species described from the banded astyanax, Astyanax fasciatus (Cuvier) in Nicaragua. Auriculostoma totonacapanensis Razo-Mendivil, Mendoza-Garfias, Pérez-Ponce de León & Rubio-Godoy, 2014 from the Mexican tetra, Astyanax mexicanus (De Filippi) in Mexico is the sister taxon of A. astyanace plus the new species. Genetic divergence levels for the 28S rDNA and ITS2 were estimated among the Middle-American species of Auriculostoma infecting characiforms. The validity of the new species is then established by reliable morphological differences, its host association to bryconids (Brycon guatemalensis Regan), restricted geographical distribution (Usumacinta and Lacantun River basins), and genetic divergence levels, albeit relatively low. A morphometric comparison between the new species and the other seven congeneric species was undertaken and, in addition, a taxonomic key to identify the species contained in the genus Auriculostoma, widely distributed across the Americas, is provided.

Morphological and molecular differentiation of Parastrigea (Trematoda: Strigeidae) from Mexico, with the description of a new species.

Parastrigea plataleae n. sp. (Digenea: Strigeidae) is described from the intestine of the roseate spoonbill Platalea ajaja (Threskiornithidae) from four localities on the Pacific coast of Mexico. The new species is mainly distinguished from the other 18 described species of Parastrigea based on the ratio of its hindbody length to forebody length. A principal component analysis (PCA) of 16 morphometric traits for 15 specimens of P. plataleae n. sp., five of Parastrigea cincta and 11 of Parastrigea diovadena previously recorded in Mexico, clearly shows three clusters, which correspond to the three species. DNA sequences of the internal transcribed spacers (ITSs) of ribosomal DNA and the mitochondrial gene cytochrome c oxidase subunit I (cox 1) were used to corroborate this morphological distinction. The genetic divergence estimated among P. plataleae n. sp., P. cincta and P. diovadena ranged from 0.5 to 1.48% for ITSs and from 9.31 to 11.47% for cox 1. Maximum parsimony (MP) and maximum likelihood (ML) analyses were performed on the combined datasets (ITSs+cox 1) and on each dataset alone. All of the phylogenetic analyses indicated that the specimens from the roseate spoonbill represent a clade with strong bootstrap support. The morphological evidence and the genetic divergence in combination with the reciprocal monophyly in all of the phylogenetic trees support the hypothesis that the digeneans found in the intestines of roseate spoonbills represent a new species.