Delimitation and species discovery in the Profundulidae fish family: Using genetic, environmental and morphologic data to address taxonomic uncertainty.

The family Profundulidae includes some one of the most enigmatic freshwater fishes of Mesoamerica: despite many attempts, a robust phylogenetic framework to delimit species is lacking, mainly due to limited morphological variation within the group. The accumulation of molecular data of profundulid fishes has led to advances in the description of new taxa, but relatively less progress has been made estimating evolutionary and phylogenetic relationships for this fish family. Here, we adopt an integrative taxonomy approach including the use of nuclear and mitochondrial DNA sequences, morphometric and ecological data, to test species boundaries in profundulid fishes in the westernmost area of their known distribution range in the states of Guerrero and Oaxaca, Mexico. Using a combination of methods for species discovery and validation based on Bayesian gene tree topologies, our analyses support the delimitation of 15 valid species of profundulid fishes - a combination of previously described species validated by this study, the synonymy of unsupported taxa, and the description of two new species. Using species delimitation methods, examination of phenotypic variation, and ecological niche characterization, we also identify five potentially new lineages which require further evidence to be erected as new species. We demonstrate that the use of an integrative taxonomy approach provides a robust methodology to delimit species in a taxonomically complex group like Profundulidae. Accurate taxonomic and ecological information is crucial for the conservation of these microendemic fishes, as several species are endangered.

Species delimitation of Gyrodactylus (Monogenea: Gyrodactylidae) infecting the southernmost cyprinids (Actinopterygii: Cyprinidae) in the New World.

The genus Gyrodactylus von Nordmann, 1832 represents one of the most diverse and widespread taxa within Monogenea, with approximately 500 species described worldwide. Thirty-three species of Gyrodactylus have been recorded in Mexico, and in the last two decades, at least 26 new species have been described mainly from freshwater fish families such as poeciliids, goodeids, profundulids, characids, and cichlids. In this study, we describe two new species of Gyrodactylus infecting freshwater cyprinids based on morphological and molecular characteristics. Gyrodactylus ticuchi n. sp. and Gyrodactylus tobala n. sp. were recovered from Notropis moralesi de Buen and N. imeldae Cortés, respectively, captured in five localities from the State of Oaxaca, Mexico. The new species differ slightly from their congeners in the morphology of the haptoral hard parts and the male copulatory organ. Sequences of the Internal Transcribed Spacers rDNA (ITS1-5.8S-ITS2), cytochrome oxidase subunit I (cox1), and the D2 + D3 domains of the large subunit (28S rDNA) were obtained from multiple specimens and analyzed using Maximum Likelihood (ML) and Bayesian Inference (BI). Phylogenetic hypotheses using ITS rDNA, cox1, and 28S rDNA genes recovered two new species of Gyrodactylus from N. moralesi and N. imeldae; we briefly discuss their phylogenetic relationship with other congeners. These gyrodactylids represent the first species described in species of Notropis from southern Mexico, the cyprinids exhibiting the southernmost distribution in the New World.

Phylogenetic relationships and ecological niche conservatism in killifish (Profundulidae) in Mesoamerica.

The family Profundulidae is a group of small-sized fish species distributed between southern Mexico and Honduras, where they are frequently the only fish representatives at higher elevations in the basins where they occur. We characterized their ecological niche using different methods and metrics drawn from niche modelling and by re-examining phylogenetic relationships of a recently published molecular phylogeny of this family to gain a better understanding of its biogeographic and evolutionary history. We assessed both lines of evidence from the perspective of niche conservatism to set a foundation for discussing hypotheses about the processes underlying the distribution and evolution of the group. In fish clades where the species composition is not clear, we examined whether niche classification could be informative to discriminate groups geographically and ecologically consistent with any of the different hypotheses of valid species. The characterization of the ecological niche was carried out using the Maxent algorithm under different parameterizations and the projection of the presence on the main components of the most relevant environmental coverage, and the niche comparison was calculated with two indices (D and I), both in environmental space and in that projected geographically. With the molecular data, a species tree was generated using the *BEAST method. The comparison of these data was calculated with an age-overlap correlation test. Based on the molecular phylogeny and on niche overlap analyses, we uncovered strong evidence to support the idea that ecologically similar species are not necessarily sister species. The correlation analysis for genetic distance and niche overlap was not significant (P > 0.05). In clades with taxonomic conflicts, we only identified Profundulus oaxacae as a geographically and ecologically distinct group from P. punctatus. All the evidence considered leads us to propose that Profundulidae do not show evidence of niche conservatism and that there are reasons to consider P. oaxacae as a valid species. Our study suggests that niche divergence is a driving evolutionary force that caused the diversification and speciation processes of the Profundulidae, along with the geological and climatic events that promoted the expansion or contraction of suitable environments.

Morphological and molecular characterization of three new species of Gyrodactylus (Monogenea: Gyrodactylidae) infecting Sicydium salvini (Teleostei: Gobiidae) in Mexican rivers draining into the Pacific Ocean.

The genus Gyrodactylus von Nordmann, 1832 is one of the most diverse within the class Monogenea; it contains mainly parasites of freshwater and marine teleost fishes. Around 40 species of Gyrodactylus have been described from gobiid fishes; mainly in Europe, as only two species are known from the Americas. In this study, we describe three new gyrodactylids from the body surface and fins of the goby Sicydium salvini (Gobiidae, Sycydiinae), which has a wide distribution on the Pacific coast, from Mexico to Panama. We describe Gyrodactylus oaxacae n. sp., G. atoyacensis n. sp. and G. salvini n. sp. collected from rivers draining to the eastern Pacific in the state of Oaxaca, Mexico. Morphologically, G. atoyacensis n. sp. and G. salvini n. sp. are very similar, and both are easily differentiated from G. oaxacae n. sp. Phylogenetic hypotheses based on sequences of the Internal Transcribed Spacers (ITS1-5.8S-ITS2 rDNA) and the D2 + D3 domains of the large ribosomal subunit (28S rDNA) support the erection of the three new taxa; and suggest that G. atoyacensis n. sp. and G. salvini n. sp. are sister species. These gyrodactylids are the first monogeneans described from gobies of the genus Sicydium in Mexico.

Morpho-molecular characterization of Gyrodactylus parasites of farmed tilapia and their spillover to native fishes in Mexico.

Translocation of fishes for aquaculture has resulted in the co-introduction of some of their parasites. African cichlid fishes, generically called "tilapias" have been introduced worldwide, along with their monogenean parasites. In a nation-wide survey, we characterised monogeneans of the genus Gyrodactylus infecting farmed "tilapia" throughout Mexico. We also collected native fishes around farms, to look for potential parasite spillover from cultured fishes. Monogeneans were identified taxonomically using morphological and molecular characters. Originally African, pathogenic Gyrodactylus cichlidarum was recorded in every farm surveyed, infecting different "tilapia" varieties, as well as three native cichlid fish species. Previously, we had shown that G. cichlidarum also infects native, non-cichlid fishes in Mexico. We also recorded that Gyrodactylus yacatli is widely distributed in Mexico, infecting cultured "tilapia" and native fishes; and present data indicating that this is a further translocated African parasite. A third, unidentified gyrodactylid infected farmed and native fishes in Chiapas, southern Mexico; we describe the new species as Gyrodactylus shinni n. sp., and provide evidence that this is a third monogenean translocated with African fish. The wide distribution of exotic parasites co-introduced with "tilapia" and their spillover to native fishes may have an important impact on the ichthyofauna in Mexico, one the world's megadiverse countries.

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.