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SUMMARY: BPA is a multifunctional endocrine disruptor with ubiquitous presence in aquatic ecosystems. The Mexican Central Plateau is an area severely impacted by pollution, inhabited by endemic viviparous fish. However, efforts to understand the effects of BPA on native species such as Goodea atripinnis are non-existent. This study focused on providing in vivo evidence of alterations in the testes of G. atripinnis males due to acute exposure to BPA at test concentrations of 1 mg/L, 10 mg/L, and 50 mg/L for 96 h. BPA exposition 1 mg/L and 10 mg/L showed degeneration and disorganization in germinal tissue. Furthermore, there was a notable decrease in sperm within the seminiferous tubules of males exposed to 10 mg/L of BPA. In all treatments, somatic cells had alterations by connective tissue thickening and an increase in collagen fibers. Additionally, inflammation and bleeding occurred in the testes of males exposed to 1 and 10 mg/L BPA. The alterations in the testes of G. atripinnis are related to BPA toxicity, which can lead to apoptosis in germ cells increasing connective tissue. Finally, even though the changes produced by BPA became evident in acute exposure (96 h), its effects are probably irreversible, compromising the reproduction of G. atripinnis.

El BPA es un disruptor endocrino multifuncional con presencia ubicua en los ecosistemas acuáticos. La Meseta Central mexicana habitada por peces vivíparos endémicos, es una zona severamente impactada por la contaminación. Sin embargo, los esfuerzos por comprender los efectos del BPA en especies nativas como Goodea atripinnis son inexistentes. Este estudio se centró en proporcionar evidencia in vivo de alteraciones en los testículos de machos de G. atripinnis debido a la exposición aguda al BPA en concentraciones de prueba de 1 mg/L, 10 mg/L y 50 mg/L durante 96 h. La exposición a BPA 1 mg/L y 10 mg/L mostró degeneración y desorganización en el tejido germinal. Además, hubo una disminución notable de los espermatozoides dentro de los túbulos seminíferos de machos expuestos a 10 mg/L de BPA. En todos los tratamientos las células somáticas presentaron alteraciones por engrosamiento del tejido conectivo y aumento de las fibras de colágeno. Además, se produjo inflamación y sangrado en los testículos de machos expuestos a 1 y 10 mg/L de BPA. Las alteraciones en los testículos de G. atripinnis están relacionadas con la toxicidad del BPA, lo que puede provocar apoptosis en las células germinales aumentando el tejido conectivo. Finalmente, si bien los cambios producidos por el BPA se hicieron evidentes en la exposición aguda (96 h), sus efectos probablemente sean irreversibles, comprometiendo la reproducción de G. atripinnis.

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Understanding the phylogeographic history of a group and identifying the factors contributing to speciation is an important challenge in evolutionary biology. The Goodeinae are a group of live-bearing fishes endemic to Mexico. Here, we develop genomic resources for species within the Goodeinae and use phylogenomic approaches to characterise their evolutionary history. We sequenced, assembled and annotated the genomes of four Goodeinae species, including Ataeniobius toweri, the only matrotrophic live-bearing fish without a trophotaenia in the group. We estimated timings of species divergence and examined the extent and timing of introgression between the species to assess if this may have occurred during an early radiation, or in more recent episodes of secondary contact. We used branch-site models to detect genome-wide positive selection across Goodeinae, and we specifically asked whether this differs in A. toweri, where loss of placental viviparity has recently occurred. We found evidence of gene flow between geographically isolated species, suggesting vicariant speciation was supplemented by limited post-speciation gene flow, and gene flow may explain previous uncertainties about Goodeid phylogeny. Genes under positive selection in the group are likely to be associated with the switch to live-bearing. Overall, our studies suggest that both volcanism-driven vicariance and changes in reproductive mode influenced radiation in the Goodeinae.

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BACKGROUND: To unravel the evolutionary history of a complex group, a comprehensive reconstruction of its phylogenetic relationships is crucial. This requires meticulous taxon sampling and careful consideration of multiple characters to ensure a complete and accurate reconstruction. The phylogenetic position of the Orestias genus has been estimated partly on unavailable or incomplete information. As a consequence, it was assigned to the family Cyprindontidae, relating this Andean fish to other geographically distant genera distributed in the Mediterranean, Middle East and North and Central America. In this study, using complete genome sequencing, we aim to clarify the phylogenetic position of Orestias within the Cyprinodontiformes order. RESULTS: We sequenced the genome of three Orestias species from the Andean Altiplano. Our analysis revealed that the small genome size in this genus (~ 0.7 Gb) was caused by a contraction in transposable element (TE) content, particularly in DNA elements and short interspersed nuclear elements (SINEs). Using predicted gene sequences, we generated a phylogenetic tree of Cyprinodontiformes using 902 orthologs extracted from all 32 available genomes as well as three outgroup species. We complemented this analysis with a phylogenetic reconstruction and time calibration considering 12 molecular markers (eight nuclear and four mitochondrial genes) and a stratified taxon sampling to consider 198 species of nearly all families and genera of this order. Overall, our results show that phylogenetic closeness is directly related to geographical distance. Importantly, we found that Orestias is not part of the Cyprinodontidae family, and that it is more closely related to the South American fish fauna, being the Fluviphylacidae the closest sister group. CONCLUSIONS: The evolutionary history of the Orestias genus is linked to the South American ichthyofauna and it should no longer be considered a member of the Cyprinodontidae family. Instead, we submit that Orestias belongs to the Orestiidae family, as suggested by Freyhof et al. (2017), and that it is the sister group of the Fluviphylacidae family, distributed in the Amazonian and Orinoco basins. These two groups likely diverged during the Late Eocene concomitant with hydrogeological changes in the South American landscape.

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Anesthetics are commonly used in fish for surgery and to facilitate capture, handling and transport in aquaculture and experimental procedures. In research, the selection of the anesthetic depends on its properties and on the recovery time. Eugenol has been pointed out as an effective anesthetic for fish, alternative to traditional drugs. Although Cnesterodon decemmaculatus is widely used as a model in ecological bioassays, no anesthetic protocol has been reported for this species. The aim of the present study was to evaluate the induction time (i.e. time to reach anesthetic stage VI) and recovery time in individuals of C. decemmaculatus subjected to eugenol at a fixed concentration, according to sex and pregnancy status. Forty-one fish were divided into three groups: males, pregnant females and non-pregnant females. They were measured for total length, standard length and weight, and the condition factor (K) was calculated. No significant differences in induction and recovery times were found for sex, pregnancy status and K between groups. Results are a contribution toward the development protocol of a standard anesthetic protocol for C. decemmaculatus.

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Abstract Introduction: Tlaloc hildebrandi is a freshwater killifish, endemic to Southern Mexico and under threat of extinction; the knowledge of the trophic morphology and diet is needed by conservation managers. Objective: To analyse and describe the anatomy of the visceral skeleton, visceral musculature, digestive tract and its adjoining glands of T. hildebrandi; as well as its diet. Methods: We performed the trophic anatomy on 20 adult specimens of both sexes, through manual dissection; as well as gut content analysis in 60 individuals to describe the diet. Results: As notable characters of the visceral skeleton of T. hildebrandi we found the posterior notch of the premaxillary, the presence of the "coronoid cartilage", the tricuspid shape of the gill rakers of the first branchial arch, and the presence of the coronomeckelian bone; some outstanding characters of the visceral musculature are the origin of the retractor dorsalis muscle from the first four vertebral centra, and the division of the pharyngoclavicularis externus muscle into two sections. The notable characters of the digestive tube are the absence of stomach and pyloric caeca, and the presence of the "intestinal valve". Insects (IVI = 66.6 %) and ostracods (13 % IVI) were the dominant prey items of the Tlaloc hildebrandi diet; larvae and adults of the family Chironomidae were the most dominant insects in the diet (53 % IVI). Conclusions: The organization of the digestive system of T. hildebrandi corresponds to the general morphologic pattern of the Cyprinodontiformes; however, we register as new information for these fish, the presence of the "coronoid cartilage" and the "intestinal valve". The structures of the trophic morphology and the components of the diet, confirms us that T. hildebrandi is a carnivorous-insectivorous fish.

Resumen Introducción: Tlaloc hildebrandi es un killi de agua dulce, endémico del sur de México y bajo amenaza de extinción; el conocimiento de la morfología trófica y la dieta son necesarios para los administradores de la conservación. Objetivo: Analizar y describir la anatomía del esqueleto visceral, la musculatura visceral, el tracto digestivo y las glándulas adyacentes de T. hildebrandi; así como los componentes de su dieta. Métodos: Mediante la técnica del descarnado manual, realizamos la descripción de la anatomía trófica en 20 especímenes adultos de ambos sexos, y el análisis del contenido estomacal en 60 individuos para describir la dieta. Resultados: Como caracteres sobresalientes del esqueleto visceral de T. hildebrandi está la escotadura posterior del premaxilar, la presencia del "cartílago coronoides", la forma tricúspide de las branquiespinas del primer arco branquial y la presencia del hueso coronomeckeliano; como caracteres de la musculatura visceral sobresalen el origen del músculo retractor dorsalis de los cuatro primeros centros vertebrales, y la división del músculo pharyngoclavicularis externus en dos secciones. Los caracteres notables del tubo digestivo son la ausencia de estómago y de ciegos pilóricos y la presencia de la "válvula intestinal". Los insectos (IVI = 66.6 %) y ostrácodos (13 % IVI) fueron los componentes dominantes de la dieta de T. hildebrandi; particularmente las larvas y adultos de la familia Chironomidae fueron los insectos más abundantes en la dieta (53 % IVI). Conclusiones: La organización del sistema digestivo de T. hildebrandi corresponde al patrón morfológico general de los Cyprinodontiformes, sin embargo, se registra como nueva información para estos peces, la presencia del cartílago coronoides y la válvula intestinal. Las estructuras de la morfología trófica y los componentes de la dieta nos confirman que T. hildebrandi es un pez carnívoro-insectívoro.

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When silica nanoparticles (SiNP) reach the water bodies interact with the already existing pollutants in the environments. This study aimed to evaluate the ecotoxicity of SiNP under the presence/absence of Cu in mosquitofish (Gambusia holbrooki). Fish were exposed to 0, 10 and 100 mg SiNP L-1, alone or mixed with Cu (0.25 mg L-1). After 96 h, the amount of colony forming units (CFU) of bacteria living on the skin mucus was analysed, and oxidative stress, tissue damage enzymes, and neurotoxicity were evaluated. We observed a reduction in CFU when Cu was present in the media. The liver was the target organ, evidencing a decrease in tissue damage enzymatic activities, activation of the antioxidant system in all treatments, and lipid oxidative damage when the SiNP and Cu were mixed. Overall, SiNP ecotoxicity was proved, which could also be enhanced by the presence of ubiquitous elements such as metals.

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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.

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In viviparous teleosts, intraovarian gestation occurs intrafollicularly, as in poeciliids, or intraluminally, as in goodeids and anablepids. Furthermore, there are two different forms of embryonic nutrition: lecithotrophy and matrotrophy; depending on the species, these can be exclusive or coexist during gestation. In matrotrophic species, nutrients are transmitted from the mother to the embryo and are especially important in species with intraluminal gestation. Jenynsia lineata is a South American viviparous teleost with intraluminal gestation, characterized by eggs with scarce yolk, which is resorbed when embryos are 6 mm long, thus developing a branchial placenta. Using histological, histochemical, and immunohistochemical techniques, the present study describes the characteristics and changes of the ovarian mucosa in J. lineata during gestational and nongestational phases, and analyzes the embryonic pharyngeal epithelium in the branchial placenta. The ovaries of 30 adult female specimens were processed using histological techniques and stained with hematoxylin-eosin, Masson's trichrome, and Alcian Blue pH 2.5/periodic acid Schiff reagent. To detect cell proliferation, we used antiproliferating cell nuclear antigen antibody. In nonpregnant females, eosinophilic granular cells (EGCs) and lymphocytes were identified in the lamina propria of the tunica mucosa, and melanomacrophage centers (MMCs) and fibroblasts were identified adjacent to tissue debris in the ovarian folds'. In the cellular debris, an embryo in resorption was observed. In pregnant females, the ovarian mucosa has thin vascularization branches entering the opercular chamber of the embryos, in close contact with the forming gill processes, thereby establishing a branchial placenta. Active cell replacement was observed in these ovarian branches. The identification of fibroblasts, lymphocytes, EGCs, and MMCs adjacent to tissue debris could indicate that these cell types are involved in the embryonic resorption process. Considering the new data obtained in this study on the branchial placenta of J. lineata, we conclude that cell proliferation could be involved in the development of maternal-embryonic interaction.

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Ibuprofen (IBP) is an anti-inflammatory drug found in aquatic environments, potentially toxic for the biota. We exposed the test fish C. decemmaculatus to two environmentally relevant concentrations (50 and 100 µg IBP/L) for 4 and 12 d and evaluated the effect on some biomarkers. Micronucleus test, nuclear abnormality test and comet assay indicated cyto-genotoxicity at both concentrations and exposure periods. Oxidative stress and biochemical biomarkers were not affected, excepting muscle AChE activity for 4 d. Muscle metabolic biomarkers showed significant decrease in ETS, lipid and protein content, while carbohydrate content was not affected. The CEA index increased at the lower IBP concentration for 4 and 12 d, possibly due to changes in body energy reserves. A full-factorial GLM performed to assess the effects of IBP and exposure times showed that the metabolic and genotoxicity biomarkers were the most sensitive to IBP toxicity, mainly at 50 µg IBP/L for 4 d.

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The mating behavior of the knife livebearer Alfaro cultratus is described in detail. During "rubbing," the male swims to a position above the female and gently moves down repeatedly touching the dorsal part of female head with the pelvic fin tips. This courtship behavior constitutes the first report of a pelvic fin male-female contact during mating in poecilids. Based on preliminary evidence, I propose that a sensory bias mechanism could mediate the evolution of signal design/mate choice in this species, which should be tested in further studies.

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