Systematic assessment of nymphs of Diesingiinae (Pentastomida: Sebekidae) parasitizing Palloceros harpagos (Cyprinodontiformes: Poeciliidae) from Parque Nacional Iguazú, Argentina: filling gaps in an incomplete phylogenetic framework.

During a search for parasites in fishes from Iguazu River, Argentina, a nymph of pentastomid was found encysted on mesenteries of Phalloceros harpagos. The 28S rDNA and COI mt-DNA were used to compare with the sequences deposited in the GenBank. Pentastomid nymphs belong to the subfamily Diesingiinae (Sebekidae) for having chloride cell pores distributed in a single row per annulus; also, the hooks and rows of spines of our material differ to other genera. Present specimens are most likely Diesingia sp., having 63-74 annuli, a U shaped oral cadre with fibers closing anteriorly and a peg like extension of the oral cadre. The 28S rDNA analysis places our samples into a sister group of Alofia sp., but the COI mt-DNA situate them on the base of the clade. In conclusion, our pentastomid positively belongs to Diesingia sp., however, indisputable assignation requires a reliable description of the nymph, or the availability of sequences linking nymphs and adults, which even could provide evidence of a new hitherto undescribed genus. Aditionally, the systematic position of Sebekia oxycephala previously described by P. harpagos is discussed on the basis of our results, allowing us to suggest a re-assignation of such specimens to the genus Diesingia.

Two new species of Gyrodactylus von Nordmann, 1832 parasitizing Cnesterodon decemmaculatus (Poeciliidae) from the southern limit of the family in the Neotropical region.

In this study, we followed an integrative taxonomy approach to describe two new species of Gyrodactylus von Nordmann, 1832, and to identify specimens of G. breviradix Vega, Razzolini, Arbetman, and Viozzi, 2019, all three collected from ten spotted live-bearer Cnesterodon decemmaculatus (Jenyns, 1842), an endemic and widespread poeciliid from the Pampean region, which is the southernmost occurring species of the Poeciliidae in the Americas. Gyrodactylids were first characterized morphologically and mophometrically, and when possible, sequences of the Internal Transcribed Spacers (ITS1-5.8S-ITS2) and the cytochrome oxidase II (COII) were used to delimit species. Gyrodactylus breviradix, Gyrodactylus marplatensis n. sp., and Gyrodactylus pampeanus n. sp. were found on the fins and body surface of C. decemmaculatus in La Tapera Creek, Mar del Plata, Buenos Aires province, Argentina. A phylogenetic analysis combining newly generated sequences of one of the new species, G. marplatensis n. sp., and of G. breviradix, along with those available in GenBank for a further 36 species of Gyrodactylus, revealed that G. marplatensis n. sp. is a sister taxon of Gyrodactylus decemmaculati Vega, Razzolini, Arbetman, and Viozzi, 2019. Genetic distances for the ITS and COII gene were estimated among Gyrodactylus spp. and further supported the validity of the new species. Overall, morphometric and molecular data coincided in delimiting the new taxa, thus demonstrating the value of integrative taxonomy for the erection of new species of Gyrodactylus and species identification.

Morphological and Molecular Evidence for a New Rhabdochonid Species (Nematoda: Rhabdochonidae) Parasitizing Profundulus oaxacae (Profundulidae) in the State of Oaxaca, Mexico.

The genus Rhabdochona Railliet, 1916 is one of the most speciose group of nematodes, parasites of freshwater fishes, with approximately 103 species described worldwide. Twenty-two species have been recorded in the Americas, 14 of them in Mexico. In this paper we describe a new species of Rhabdochona on the basis of light and scanning electron microscopy and molecular tools. Rhabdochona adentata n. sp. was recovered from the gallbladder of the freshwater Oaxaca killifish, Profundulus oaxacae (Meek, 1902) (Profundulidae) captured in the Río Grande, State of Oaxaca, Mexico. Rhabdochona adentata n. sp. differs markedly from its congeners by possessing an unusual prostom lacking anterior teeth, small simple deirids, and the location of the excretory pore at the level of the union of the muscular and glandular esophagus. Sequences of cytochrome c oxidase subunit I (COI) from mitochondrial deoxyribonucleic acid and the D2 + D3 domains of the large ribosomal subunit (28S) were obtained from 3 specimens and were analyzed using maximum likelihood and Bayesian inference analyses. Phylogenetic analyses using COI and 28S genes recovered 1 new lineage of Rhabdochona. The new species is described on the basis of a detailed morphological study. This parasite represents the first species of Rhabdochona without prostomal teeth and with a different site of infection, the gallbladder.

The role of freshwater fish in the life cycle of Dioctophyme renale in Southern Brazil.

Brazil stands out by the diverse records of Dioctopphyme renale in different hosts; however, there is little information about the life cycle of the nematode in the region. This study aims to report on third-stage larvae infections in fish in southern Brazil. In this context, 324 fish of different species belonging to Characiformes, Cyprinodontiformes and Siluriformes were collected in an urban area of Rio Grande do Sul State, where domestic and wild hosts were reported with the nematode. Of the total fish examined, 25(7.7%) were found to be parasitized by third-stage larvae of D. renale which were found encysted in the stomach serous membrane and in the celoma cavity of Hoplosternum littorale (Siluriformes) with a prevalence of 53.2% (25/47) and mean intensity of infection of 4.4 larvae/host (1 to 13 larvae). The occurrence of larvae in H. littorale indicates the presence of parasitosis in the region; however, the contribution of this fish species as a source of infection for dogs in urban areas must be considered with caution given the difficulties these dogs may face in the capture and predation of the fish to the point of effectively maintaining the urban cycle of D. renale. In addition, the low level of larvae registered in the total sample of fish examined indicates that these hosts are unlikely to play an important role in the transmission of D. renale to domestic animals in the region of the study.

Dendrorchis pampae sp. n. (Digenea: Gorgoderidae) from Cynopoecilus melanotaenia (Cyprinodontiformes: Cynolebiidae) a killifish from southern Brazil, with an emended diagnosis of Dendrorchis.

A new species of Dendrorchis is described and compared with others in the genus. The parasites were found in the swim bladder of the annual killifish Cynopoecilus melanotaenia. Hosts were collected from a seasonal wetland in southern Brazil. The main characteristics of D. pampae are: genital pore in the intestinal bifurcation region elongate and lobed vitellaria uterine loops limited to the acetabular region and to the rear end of the body; and wide intestinal caeca. An emended diagnosis of the genus Dendrorchis includes the characteristics of the new species. This is the first record of an adult digenean in an annual killifish from South America.

Two new species of Gyrodactylus von Nordmann, 1832 from Profundulus oaxacae (Pisces: Profundulidae) from Oaxaca, Mexico, studied by morphology and molecular analyses.

In the present study, two new species of Gyrodactylus are described from Profundulus oaxacae, a fish endemic to the Pacific slope of Oaxaca State, Mexico. Fishes were collected within their distribution range in 5 localities in the Atoyac-Verde River. Gyrodactylus montealbani n. sp. and G. zapoteco n. sp. were erected and characterized morphologically (sclerites of the attachment apparatus and the male copulatory organ) and molecularly (sequences of the Internal Transcribed Spacer region of rDNA). The haptoral sclerites of the new species are similar to those of Gyrodactylus iunuri and Gyrodactylus tepari, both recently described from the goodeid fish Goodea atripinnis, from the Mexican States of Jalisco and Querétaro, respectively; and to Gyrodactylus xtachuna described from the poeciliid Poeciliopsis gracilis in Veracruz State, Mexico - nonetheless, these species can all be discriminated based on their marginal hook morphology. Specimens of G. montealbani n. sp. and G. zapoteco n. sp. were sequenced, and were aligned with sequences of 25 other Gyrodactylus spp. Both Maximum likelihood and Bayesian inference analyses indicated that the two new species are members of independent, well-supported lineages - these are the first Gyrodactylus species described from Profundulus oaxacae.

Three new species of Gyrodactylus von Nordmann, 1832 described from Goodea atripinnis (Pisces: Goodeidae), an endemic freshwater fish from the central highlands of Mexico.

Goodea atripinnis Jordan, 1880 has a broad range of habitats and is the most widespread species of the endemic goodeid fishes, which inhabit the central highlands of Mexico. This species is known to be host to a high diversity of helminth parasites from which only three belong to the genus Gyrodactylus von Nordmann, 1832: G. lamothei Mendoza-Palmero, Sereno-Uribe et Salgado-Maldonado, 2009, G. mexicanus Mendoza-Palmero, Sereno-Uribe et Salgado-Maldonado, 2009, and G. tomahuac Rubio-Godoy, Razo-Mendivil, García-Vásquez, Freeman, Shinn et Paladini, 2016. Here, we describe three new species of Gyrodactylus collected from G. atripinnis, which were characterised morphologically (sclerites of the attachment apparatus) and molecularly (sequences of the internal transcribed spacer region of the rDNA): Gyrodactylus iunuri n. sp., Gyrodactylus katamba n. sp. and Gyrodactylus tepari n. sp. These new species were collected in three different states in the Mexican Highlands: Guanajuato, Jalisco and Querétaro. Both morphological and molecular data support the hypothesis that two distinct groups of gyrodactylids infect goodeid fishes: G. iunuri n. sp., G. tepari n. sp. and G. tomahuac possess robust hamuli and are closely related phylogenetically; while G. katamba n. sp. resembles G. lamothei in having slender hamuli with accessory sclerites adjacent to the hamuli root, and apparently shares a common ancestor with gyrodactylids infecting poeciliid fishes. New locality records of G. tomahuac are presented. The addition of the three new species of Gyrodactylus as parasites of G. atripinnis makes monogeneans the second most abundant parasite group known to infect this host.

First Freshwater Bothriocephalidean (Cestoda) from Tropical South America, Closely Related to African Taxa.

Regobothrium microhamulinum n. gen. and n. sp. (Bothriocephalidea: Bothriocephalidae) is proposed to accommodate a new cestode from flatfish Catathyridium jenynsii (Chabanaud, 1922) (Pleuronectiformes: Acharidae; type host) and another 3 freshwater fishes of the orders Characiformes, Cyprinodontiformes, and Siluriformes in the Neotropical Region. The new genus is placed in the Bothriocephalidae because it possesses medioventral uterine and mediodorsal genital pores and a follicular vitellarium. Regobothrium n. gen. is characterized by possessing a tiny, slightly subovate scolex narrower than the strobila, with an apical disc armed with 2 semicircles of 15-17 tiny hooks in each and an acraspedote strobila. Regobothrium n. gen. differs from all bothriocephalid cestodes that have a scolex armed with hooks by their small size (maximum length less than 20 µm) and a triangular shape with the basal part (handle or basal plate) shorter than the distal coniform part (blade). In the other hooked bothriocephalids, hooks have a longer handle than a blade. Regobothrium microhamulinum n. gen. and n. sp. is the third bothriocephalidean cestode described from freshwater teleosts in South America but the first out of Patagonia. Molecular phylogenetics consider Regobothrium as a member of a lineage consisting of (up to now exclusively) freshwater bothriocephalids from the Ethiopian biogeographic region, thus indicating Gondwanan relationship.

Short communication: first report of nematodes parasitizing the four-eyed-fish, Anableps anableps (Pisces, Cyprinodontiformes).

The genus Anableps is composed of species Anableps anableps, Anableps dowi, and Anableps microlepis. These fishes are tropical and usually live on the surface of brackish water, being popularly known as four-eyed-fishes due to the presence of prominent eyes and a pupil split horizontally. A. anableps and A. microlepis are considered as sister species that live in sympatry in South America. A. dowi, however, is restricted to the Pacific Ocean (Central America) and is considered the most primitive species of this genus. The aims of this study were to investigate the presence of endoparasites in A. anableps from the Parnaíba's Delta and characterize them morphologically. During the necropsy, larvae of Contracaecum sp. in the third larval stage (L3) were collected from the pancreas of A. anableps, but no endoparasites were observed in other organs. The worms had a cuticular tooth and excretory pore located at the anterior end, a thread like body, whitish color, and without distinction of sex. The length of the ventricular appendix of the larvae was much greater than in other studies. This is the first report of endoparasitism in A. anableps and the first report of nematodes in four-eyed-fishes.

Molecular, Morphological, and Ecological Data of Saccocoelioides Szidat, 1954 (Digenea: Haploporidae) from Middle America Supported the Reallocation from Culuwiya cichlidorum to Saccocoelioides.

This study investigates the systematic position of some species of the genus Saccocoelioides Szidat, 1954, and the species Culuwiya cichlidorum Aguirre-Macedo and Scholz, 2005 , from North and Middle America using new morphological data and partial sequences of 2 nuclear genes, the large subunit and the internal transcribed spacer 2 from ribosomal DNA. In total 74 specimens representing 4 species of Saccocoelioides (41 of Saccocoelioides sogandaresi, 3 of Saccocoelioides chauhani, 6 of Saccocoelioides lamothei, and 12 of Saccocoelioides olmecae) plus 12 specimens of Culuwiya cichlidorum were analyzed. The phylogenetic analyses inferred with maximum likelihood method and Bayesian inference showed that the 4 species of Saccocoelioides formed a clade divided into 4 subclades representing each species. In addition, 7 specimens sequenced from the type host and locality of Culuwiya cichlidorum were nested in 2 clades: the first clade with specimens of C. cichlidorum from Costa Rica, and the second clade with specimens of S. sogandaresi from Mexico, Honduras, Nicaragua, and Costa Rica. Our analyses suggest that C. cichlidorum should be reallocated into Saccocoelioides, as was originally designated Saccocoelioides cichlidorum n. comb., is restricted to Nicaragua and Costa Rica and associated with cichlid fishes. In contrast, the specimens identified previously as Culuwiya cichlidorum from the coast of Gulf of Mexico by Aguirre-Macedo and Scholz belong to S. sogandaresi, which has a wide distribution that extends from Galveston Bay (U.S.A.) in Gulf of Mexico to Atlantic and Pacific slopes associated with poeciliid and cichlid fishes. New morphological data derived from scanning electron microscopy photographs of the body surface show a characteristic pattern of spination that is useful to distinguish between S. cichlidorum n. comb. and S. sogandaresi.

Triple trouble? Invasive poeciliid fishes carry the introduced tilapia pathogen Gyrodactylus cichlidarum in the Mexican highlands.

As part of ongoing surveys of the gyrodactylid parasite fauna of freshwater fishes in Mexico, we recorded the infection of three species of poeciliids (Poecilia mexicana, Poeciliopsis gracilis, and Pseudoxiphophorus bimaculatus [syn.=Heterandria bimaculata]) with Gyrodactylus cichlidarum, a monogenean parasite of cichlid fishes, which has been co-introduced globally with its translocated, African "tilapia" hosts. This tilapia pathogen was found on poeciliid fishes both within their native distribution range in the Gulf of Mexico slope, as well as on invasive species artificially introduced to the Mexican highlands, to rivers draining into the Pacific Ocean. Identity of G. cichlidarum was confirmed by morphological and molecular analyses. Prevalence and abundance of infection were low, but this is the first record of G. cichlidarum infecting poeciliids (Cyprinodontiformes), which are distantly related to this parasite's primary cichlid fish hosts (Perciformes). This study provides evidence that G. cichlidarum, a recognized pathogen which has been co-introduced globally with its cichlid fish hosts for aquacultural purposes, is able to infect non-related poeciliid fishes inhabiting water bodies adjacent to tilapia farms, thereby potentially increasing its ability to disperse between farms and different river basins. It is of particular concern that G. cichlidarum was found on poeciliids, as these invasive fishes have been introduced worldwide and could act as carriers for this parasite known to induce significant mortality of farmed tilapias - globally, the second most important freshwater aquaculture fish group, after the carps.

To each his own: no evidence of gyrodactylid parasite host switches from invasive poeciliid fishes to Goodea atripinnis Jordan (Cyprinodontiformes: Goodeidae), the most dominant endemic freshwater goodeid fish in the Mexican Highlands.

BACKGROUND: Goodeid topminnows are live-bearing fishes endemic to the Mexican Highlands (Mesa Central, MC). Unfortunately, in the MC, environmental degradation and introduced species have pushed several goodeid species to the brink of extinction. Invasive fishes can introduce exotic parasites, and the most abundant goodeid, blackfin goodea Goodea atripinnis Jordan, is parasitised by six exotic helminths. Poeciliids are widely dispersed invasive fishes, which exert negative ecological effects on goodeids. Poeciliids host several species of the monogenean genus Gyrodactylus von Nordmann, 1832, including pathogenic, invasive parasites. Here, we looked for evidence of Gyrodactylus species switching hosts from poeciliids to goodeids. METHODS: Fish were collected in rivers draining the MC into both sides of the continental divide. Hosts were screened for gyrodactylid parasites in localities where G. atripinnis and poeciliids occurred sympatrically. Gyrodactylus specimens were characterised morphologically (attachment apparatus) and molecularly (internal transcribed spacer region, ITS). A Bayesian phylogenetic tree using ITS sequences established relationships between gyrodactylids collected from goodeid fishes and those from parasites infecting poeciliids. RESULTS: Gyrodactylids were collected from G. atripinnis in six localities on both sides of the watershed where exotic poeciliids occurred sympatrically. Morphological and molecular analyses indicated the presence of four undescribed species of Gyrodactylus infecting this goodeid host. Gyrodactylus tomahuac n. sp., the most abundant and geographically widespread species, is described here. The other three Gyrodactylus spp. are not described, but their ITS sequences are used as molecular data presented here, are the only available for gyrodactylids infecting goodeid fishes. Morphological and molecular data suggest that two distinct groups of gyrodactylids infect goodeids, one of which shares a common ancestor with gyrodactylids parasitizing poeciliids. CONCLUSIONS: No evidence was found of gyrodactylids switching hosts from invasive poeciliids to endemic goodeids, nor vice versa. Moreover, considering that G. atripinnis is known to host both Gyrodactylus lamothei Mendoza-Palmero, Sereno-Uribe & Salgado-Maldonado, 2009 and Gyrodactylus mexicanus Mendoza-Palmero, Sereno-Uribe & Salgado-Maldonado, 2009, with the addition of G. tomahuac n. sp. and the three undescribed Gyrodactylus spp. reported, at least six gyrodactylids may infect this host. This would make monogeneans the second most abundant parasite group infecting G. atripinnis, which to date is known to harbour 22 helminth species: nine digeneans, five nematodes, four cestodes, three monogeneans and one acanthocephalan.

Phylogenetic Analysis Using the 28S rRNA Gene Reveals That the Genus Paracreptotrema (Digenea: Allocreadiidae) Is Not Monophyletic; Description of Two New Genera and One New Species.

This study investigates the systematics of Paracreptotrema Choudhury, Pérez-Ponce de León, Brooks and Daverdin, 2006 using morphological data (stained whole mounts and scanning electron microscopy) and partial sequences of the 28S ribosomal rRNA gene, obtained from freshly collected material. In total, 484 specimens representing 4 species, i.e., Paracreptotrema blancoi (157), Paracreptotrema profundulusi (12), Paracreptotrema rosenthali (8), and Paracreptotrema blancoi sensu Salgado-Maldonado et al. (2011) (307) were collected. Existing museum depositions were also studied. The 28S rRNA gene sequences of these Paracreptotrema spp. were aligned, along with sequences from 22 other allocreadiids and 4 other non-allocreadiid xiphidiatan species. Bayesian inference and maximum likelihood analyses indicated a paraphyletic Paracreptotrema split into 3 clades: 1 comprising P. blancoi and P. rosenthali that was sister to a clade formed by 3 other species of allocreadiids (species of Wallinia, Creptotrematina, and Auriculostoma) typically found in characid fishes, a second clade formed solely by Paracreptotrema heterandriae as the sister taxon of the aforementioned species, and a third by P. profundulusi and specimens erroneously identified as P. blancoi. Two new taxa were erected to reflect these results: Paracreptotrematoides for Paracreptotrema heterandriae, and Pseudoparacreptotrema for Paracreptotrema profundulusi and P. macroacetabulata (the species erroneously identified as P. blancoi from profundulids across Middle America). Closer consideration of the morphology corroborates these findings. The revised systematics also indicated that Paracreptotrema spp. are found in poeciliids, whereas Pseudoparacreptotrema spp. parasitize profundulids. The study demonstrates the value of an integrative taxonomy approach to address the apparently complicated systematics of the allocreadiids.

Two new species of Phyllodistomum Braun, 1899 (Digenea: Gorgoderidae), from freshwater fishes (Cyprinodontiformes: Goodeidae: Goodeinae) in central Mexico: An integrative taxonomy approach using morphology, ultrastructure and molecular phylogenetics.

An integrative taxonomy approach is used to characterise the diversity of gorgoderid trematodes that parasitize freshwater fishes of the subfamily Goodeinae in central Mexico. Records of Phyllodistomum sp. and Dendrorchis sp. from the urinary bladder of goodeines have been previously published, although the identification at species level was not achieved. A few specimens were collected and fixed to conduct a scanning electron microscopy study, and to obtain sequences of a mitochondrial (COI) and nuclear (28S rRNA) gene, to be analysed in the context of the molecular phylogeny of gorgoderid trematodes. Based on the new findings, two new species of Phyllodistomum Braun, 1899 are described. Phyllodistomum cribbi n. sp. was found in Zoogoneticus quitzeoensis (Bean), Allotoca zacapuensis Meyer, Radda & Domínguez-Domínguez, Hubbsina turneri de Buen and Z. purhepechus Domínguez-Domínguez, Pérez-Rodríguez & Doadrio from Zacapu Lake, and La Luz Spring, in Michoacan, central Mexico. Phyllodistomum wallacei n. sp. parasitized Xenotaenia resolanae Turner, Ilyodon furcidens (Jordan & Gilbert), and Allodontichthys tamazulae Turner from the Cuzalapa, Ayuquila and Tamazula Rivers in Jalisco, western Mexico. These species are compared with several freshwater Phyllodistomum species from different areas of the world, especially a group of eight species that comprise a monophyletic clade in recent phylogenetic hypotheses of the Gorgoderidae Looss, 1899. The two new species are distinguished from other close relatives by the combination of morphological traits such as the body shape, sucker ratio, shape of the gonads, and extension of intestinal ceca. The new species are distinct in some ultrastructural characters of the body surface when compared with those species where scanning electron micrographs (SEM) and/or microphotographs are available. Data of two molecular markers (28S rRNA and COI genes) demonstrate that the two new species are distinct from each other and from those species of Phyllodistomum Braun, 1899 for which sequences are available.

Phyllodistomum spinopapillatum sp. nov. (Digenea: Gorgoderidae), from the Oaxaca killifish Profundulus balsanus (Osteichthyes: Profundulidae) in Mexico, with new host and locality records of P. inecoli: Morphology, ultrastructure and molecular evidence.

Phyllodistomum spinopapillatum sp. nov. is described from the urinary bladder of the Oaxaca killifish, Profundulus balsanus Ahl (Profundulidae) in Rio Pueblo Viejo and Rio Santa Cruz, Oaxaca, southwestern Mexico. The new species is described based on evidence gathered from morphology, scanning electron microscopy (SEM), and partial sequences of the 28S rRNA gene. Diagnostic characters of the new species of Phyllodistomum Braun 1899 are the presence of spines on the entire body surface and having a ventral sucker almost half the size of oral sucker. The new species possess a large number of dome-like papillae on the body surface with scattered distribution along the hindbody, and these papillae are characteristically spinulated. Phyllodistomum spinopapillatum sp. nov. most closely resembles P. inecoli Razo-Mendivil, Perez-Ponce de Leon and Rubio- Godoy, 2013, a parasite of the twospot livebearer Pseudoxiphophorus bimaculatus (Heckel) from Veracruz, in the Atlantic slope of Mexico. In addition to the new species, specimens of P. inecoli were also found parasitising the urinary bladder of cyprinodontiforms such as the Mexican molly Poecilia sphenops Valencienes in a pond at Santa Maria Coyotepec, and in Profundulus sp. in Rio Templo, both in Oaxaca, and in the Porthole livebearer Poeciliopsis gracilis (Heckel) in Rio San Juan, as well as in Profundulus punctatus (Gunter) from Rio Nueva Francia, both in Chiapas. The distribution and host range of P. inecoli is extended to freshwaters of the Pacific slope of Mexico, and to other cyprynodontiforms.

Coevolution between Contracaecum (Nematoda, Anisakidae) and Austrolebias (Cyprinodontiformes, Rivulidae) host-parasite complex from SW Atlantic coastal basins.

In recent years, molecular studies in host-parasite interactions in terms of coevolution have become important. Larvae (L3) of two species of Contracaecum were found parasitizing species of Rivulidae in the Atlantic coastal basins from Uruguay. The aim of this study is to determine the patterns of differentiation of this host-parasite complex in order to clarify possible coevolutionary events in such interaction throughout phylogeographic approach using both nuclear and mitochondrial molecular markers (internal transcribed spacers (ITS) and cytochrome oxidase subunit 1 (cox-1)). Based on both markers, intraspecific variation in Contracaecum species was lower than 2 %, while interspecific variation was greater than 10 %. Both species of Contracaecum constitute monophyletic groups. Contracaecum resulted in a paraphyletic genus when incorporating other Contracaecum species and closely related nematode sequences from GenBank. ITS regions showed that Contracaecum sp. 1 is more closely related to other species of the same genus than with their counterparts from Atlantic coastal basins in Uruguay. Haplotype network for both markers corroborate the existence of two distinct taxa. While ITS pairwise FST comparisons and the indirect estimate of gene flow confirm the existence of two distinct Contracaecum species, mitochondrial gene detected low levels of migrants between some of the populations from both species. Our results suggest that coevolution in this host-parasite complex species is plausible. Parasite cladogenetic events occur almost simultaneously with the separation of the hypothetical ancestors of each species complex of Austrolebias during Pliocene. Additionally, the two lineages of Contracaecum colonize differently the species within each of the Austrolebias complexes.

Checklist of helminth parasites of Goodeinae (Osteichthyes: Cyprinodontiformes: Goodeidae), an endemic subfamily of freshwater fishes from Mexico.

From August 2008 to July 2010, 1,471 fish belonging to the subfamily Goodeinae (representing 28 species) were collected from 47 localities across central Mexico and analyzed for helminth parasites. In addition, a database with all available published accounts of the helminth parasite fauna of goodeines was assembled. Based on both sources of information, a checklist containing all the records was compiled as a necessary first step to address future questions in the areas of ecology, evolutionary biology and biogeography of this host-parasite association. The checklist is presented in two tables, a parasite-host list and a host-parasite list. The checklist contains 51 nominal species, from 34 genera and 26 families of helminth parasites. It includes 8 species of adult digeneans, 9 metacercarie, 6 monogeneans, 3 adult cestodes, 9 metacestodes, 1 adult acanthocephalan, 1 cystacanth, 6 adult nematodes and 8 larval nematodes. Based on the amount of information contained in the checklist, we pose that goodeines, a subfamily of viviparous freshwater fishes endemic to central Mexico, might be regarded as the first group of wildlife vertebrate for which a complete inventory of their helminth parasite fauna has been completed.

Do the historical biogeography and evolutionary history of the digenean Margotrema spp. across central Mexico mirror those of their freshwater fish hosts (Goodeinae)?

Host-parasite systems provide an ideal platform to study evolution at different levels, including codivergence in a historical biogeography context. In this study we aim to describe biogeographic and codivergent patterns and associated processes of the Goodeinae freshwater fish and their digenean parasite (Margotrema spp.) over the last 6.5 Ma (million years), identifying the main factors (host and/or hydrogeomorphology) that influenced the evolution of Margotrema. We obtained a species tree for Margotrema spp. using DNA sequence data from mitochondrial and nuclear molecular markers (COI and ITS1, respectively) and performed molecular dating to discern divergence events within the genus. The dispersal-extinction-cladogenesis (DEC) model was used to describe the historical biogeography of digeneans and applied to cophylogenetic analyses of Margotrema and their goodeine hosts. Our results showed that the evolutionary history of Margotrema has been shaped in close association with its geographic context, especially with the geological history of central Mexico during the Pleistocene. Host-specificity has been established at three levels of historical association: a) Species-Species, represented by Xenotaenia resolanae-M. resolanae exclusively found in the Cuzalapa River Basin; b) Species-Lineage, represented by Characodon audax-M. bravoae Lineage II, exclusive to the Upper and Middle Mezquital River Basin, and c) Tribe-Lineage, including two instances of historical associations among parasites and hosts at the taxonomical level of tribe, one represented by Ilyodontini-M. bravoae Lineage I (distributed across the Ayuquila and Balsas River Basins), and another comprised of Girardinichthyini/Chapalichthyini-M. bravoae Lineage III, found only in the Lerma River Basin. We show that the evolutionary history of the parasites is, on several occasions, in agreement with the phylogenetic and biogeographic history of their hosts. A series of biogeographic and host-parasite events explain the codivergence patterns observed, in which cospeciation and colonisation via host-switching and vicariant plus dispersal events are appreciated, at different times during the diversification history of both associates, particularly during the Pleistocene.

Testing the systematic position and relationships of Paracreptotrema heterandriae within the Allocreadiidae through partial 28s rRNA gene sequences.

The genus Paracreptotrema Choudhury, Pérez-Ponce de León, Brooks and Daverdin, 2006 (Digenea), comprises 4 species that parasitize poeciliid and cyprinodontid fishes in Middle America. Based on morphological characters exhibited by adults of the type species (Paracreptotrema blancoi Choudhury, Pérez-Ponce de León, Brooks and Daverdin, 2006 ) the genus was tentatively assigned to Allocreadiidae Looss, 1902. We sequenced domains D1-D3 of the 28S rRNA gene of several specimens of Paracreptotrema heterandriae Salgado-Maldonado, Caspeta-Mandujano and Vázquez, 2012 , and analyzed these in a phylogenetic context along with 10 allocreadiid taxa plus several xiphidiatan and 2 monorchiatan digeneans, in order to test the proposed inclusion of P. heterandriae in Allocreadiidae. Maximum likelihood and Bayesian inference analyses of 28S rRNA gene sequences confirmed the classification of Paracreptotrema as an allocreadiid. In the phylogenetic trees, Paracreptotrema appears to be the sister taxon of Creptotrematina aguirrepequenoi Jiménez-Guzmán, 1973, and both are closely related to Auriculostoma Scholz, Aguirre-Macedo, and Choudhury, 2004, with Wallinia Pearse, 1920, as the sister taxon of all these genera; nevertheless, the interrelationships between these 4 genera were not totally resolved. ITS2 sequences of several specimens of P. heterandriae showed null intraspecific variation. Scanning electron microscopy microphotograph demonstrated the presence of 13 and 4 dome-like papillae arranged around the outer and inner edges of the oral sucker, respectively.

Integrative taxonomy identifies a new species of Phyllodistomum (Digenea: Gorgoderidae) from the twospot livebearer, Heterandria bimaculata (Teleostei: Poeciliidae), in Central Veracruz, Mexico.

Phyllodistomum inecoli n. sp. is described from the twospot livebearer, Heterandria bimaculata (Teleostei: Poeciliidae), collected in the Río La Antigua basin, Veracruz, Mexico. The new species is described and characterised by using a combination of morphology, scanning electron microscopy, and sequences of nuclear and mitochondrial genes. Diagnostic characters of the new species of Phyllodistomum include a genital pore opening at the level of the caecal bifurcation; oval vitellarium, situated just posterior to the ventral sucker and not extended laterally and anterior extracaecal uterine loops variable in extension (reaching the anterior, median or posterior margin of the ventral sucker). P. inecoli n. sp. most closely resembles P. brevicecum, a species described as a parasite of the central mudminnow, Umbra limi, in other parts of North America; however, the genital pore in P. brevicecum is situated between the caecal bifurcation and the ventral sucker, the ovary is larger, the vitellarium is lobed and extended laterally and the anterior portion of the uterus extends to the posterior margin of the ventral sucker. Comparison of about 1,500­2,200 nucleotides of cox1 and 28S rDNA and ITS1 strongly supports the status of P. inecoli as a new species. Bayesian inference analysis of combined datasets of 28S rDNA and cox1 sequences showed that P. inecoli n. sp. and the other species found in freshwater fishes of Mexico, including the species complex of P. lacustri, are not sister species. Phylogenetic analysis based on 28S rDNA sequences of several gorgoderid taxa revealed the close relationship of P. inecoli n. sp. with several species of Phyllodistomum, Gorgodera and Gorgoderina with cystocercous cercariae developing in sphaeriid bivalves. Dot-plot analysis of ITS1 sequences of P. inecoli n. sp. revealed the presence of eight repetitive elements with different length, which together represent almost half the length of ITS1.