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The teleost Astyanax mexicanus consists of surface dwelling (surface fish) and cave dwelling (cavefish) forms. Cavefish have evolved in subterranean habitats characterized by reduced oxygen levels (hypoxia) and exhibit a subset of phenotypic traits controlled by increased Sonic hedgehog (Shh) signaling along the embryonic midline. The enhancement of primitive hematopoietic domains, which are formed bilaterally in the anterior and posterior lateral plate mesoderm, are responsible for the development of more larval erythrocytes in cavefish relative to surface fish. In this study, we determine the role of hypoxia and Shh signaling in the development and evolution of primitive hematopoiesis in cavefish. We show that hypoxia treatment during embryogenesis increases primitive hematopoiesis and erythrocyte development in surface fish. We also demonstrate that upregulation of Shh midline signaling by the Smoothened agonist SAG increases primitive hematopoiesis and erythrocyte development in surface fish, whereas Shh downregulation via treatment with the Smoothened inhibitor cyclopamine decreases these traits in cavefish. Together these results suggest that hematopoietic enhancement is regulated by hypoxia and Shh signaling. Lastly, we demonstrate that hypoxia enhances expression of Shh signaling along the midline of surface fish embryos. We conclude that hypoxia-mediated Shh plasticity may be a driving force for the adaptive evolution of primitive hematopoiesis and erythrocyte development in cavefish. [Display omitted] • Hypoxia increases hematopoiesis and erythrocytes in surface fish. • Shh upregulation increases hematopoiesis and erythrocytes in surface fish. • Shh inhibition decreases hematopoiesis and erythrocytes in cavefish. • Hypoxia upregulates Shh along the embryonic midline in surface fish. [ABSTRACT FROM AUTHOR]

Charles Breder, a pioneering researcher of blind Mexican cavefish was the first to note extreme variation in the facial skeleton of this intriguing subterranean-dwelling organism. Using a system of polar coordinate plots, he identified substantial dysmorphic changes affecting bones of the orbital skeleton. A complication of his landmark publication from 1944 was an error in the number of orbital bones depicted for this species. Intriguingly, however, he proposed an unknown "organizing force" likely influences final bone position and associated dysmorphia. At the time this was merely hypothetical. Roughly eight decades since its publication, however, insights into sensory influences on facial bone development may explain dysmorphia and variation in bone numbers for Astyanax cavefish. A morphological association between mechano-sensory neuromasts of the lateral line and dermal bones of the facial skeleton had been appreciated in the classical literature, but the polarity of this interaction has long remained unclear. Here, we propose that sensory-skeletal integration between sensory neuromasts and bones explain the incomplete numbers of bones, and dysmorphic features such as fusion between neighboring elements. We propose that in closely-related surface fish (and most teleost fish) this developmental coupling enables the sensory and skeletal systems to become integrated into a functional unit over the course of life history. In this opinion article, we discuss the relevance of this (poorly understood) phenomenon as a potential evolutionary source of variation in the facial bone structures of taxa across deep geologic time. We provide three potential explanations for the error in Breder's drawings, that may be explained by natural developmental variation documented in other related species. Moreover, we argue that the natural variation in this "evolutionary" model system is useful for explaining diverse cranial features by uniting aberrations occurring during embryogenesis with long-term adult dysmorphia. [Display omitted] • Charles Breder represented Astyanax as having too few orbital bones. • Breder proposed an "unknown organizing force" governed development of these bones. • This "force" is likely the osteogenic capacity of sensory neuromasts. • Sensory neuromasts likely induce bone to prefigure their functional role. • Variations on sensory-skeletal integration explain aberrations in cavefish. [ABSTRACT FROM AUTHOR]

Triploid induction is a promising biotechnique that could be used to enhance aquaculture yields in the near future. However, studies conducted with several fish species have demonstrated that the presence of an extra set of chromosomes may result in deleterious health effects. Furthermore, studies of fish immune responses still need to be conducted before these specimens can be readily commercialized. In the study presented herein, we evaluated the effects of triploid induction on hematology, erythrocyte morphometry and morphology, phagocytosis, and the expression levels of IL-1β and TGF-β using specimens of the Neotropical species, Astyanax altiparanae. In general, the cell counts of erythrocytes, leukocytes, and neutrophils in triploid fish were lower than those in diploid fish. The erythrocytes of triploid fish were larger than those found in diploid fish, but also demonstrated considerably higher frequencies of cellular and nuclear abnormalities. Although not statistically significant, triploid induction resulted in a phagocytic capacity (PC) 20% lower than that found with diploid fish. No notable differences were observed in phagocytic index (PI). Gene expression levels for the cytokine IL-1 were lower in tissues from the head kidney, liver, and spleen of triploid fish with respect to diploid fish. Gene expression levels of TGF-β were lower only in the spleen of triploids compared to diploids. In conclusion, triploid induction resulted in A. altiparanae specimens with immune impairments and potentially lower resistances to disease and low-quality environments. • First study regarding triploid induction and phenotypical and molecular immune response of a Neotropical teleost. • Triploid induction decreases neutrophil numbers in A. altiparnae. • Triploid induction decreases erythrocytes numbers and cause erythrocytic deformities. • Triploid induction causes mild impact on phagocytosis in A. altiparnae. • Triploid induction decreases expression of IL-1β and TGF- β in A. altiparnae. [ABSTRACT FROM AUTHOR]

• Males from pollutes sites had high levels of E2 and lower T and 11-KT. • Females displayed higher 11-KT but T, E2 and 17-OHP were not changed. • Proportion of the germ cells was changed in males and females. • Gonad histopathologies and oocyte ageing were also found. Discharge of municipal wastewater promotes the entry of diverse oestrogenic compounds into the water bodies. This complex mixture of substances interferes in the steroidogenic pathway, being able to promote severe reproductive impairment in freshwater fish populations. The purpose of the present study was to evaluate the effects of oestrogenic endocrine disruptors (EDCs) mixture on gonadal sex steroids (testosterone, T; 11-ketotestosterone, 11-KT; 17β-oestradiol, E2; 17-hydroxyprogesterone, 17-OHP) in the peak of the reproductive season of Astyanax rivularis , correlating the results obtained with the proportion of germ cells and gonadal histopathology. Three sampling sites were chosen to conduct the study, one reference site (S1), without contamination by municipal wastewater and two sites (S2 and S3) receiving discharge of municipal wastewater. Males of A. rivularis presented higher concentrations of E2, lower androgens (T and 11-KT) in gonads when compared to males from site S1. Concentrations of 17-OHP did not present significant difference among sites. In sites S2 and S3, the proportion of early spermatocytes, spermatids and Leydig cells increased while spermatozoa decreased compared to fish from S1. The following gonadal histopathologies were detected in the male fishes: intersex gonads (28% in S3) and testicular degeneration with germinal epithelium exhibiting agglutinated germ cells masses and empty cysts (57% in S2 and 71% in S3). In females, concentrations of T, E2 and 17-OHP did not present significant difference among the sites, however higher 11-KT concentrations were detected in females from sites S2 and S3. A lower proportion of perinucleolar follicles and a higher incidence of vitellogenic follicles, besides, aged oocytes and the presence of eosinophilic proteinaceous fluid in the interstitial compartment were also found in females from impacted sites. These results indicate that the urbanization and consequent release of municipal wastewater containing oestrogenic compounds in the headwater creeks are altering the levels of sex hormones and gametogenesis of A. rivularis. Further studies should be performed to determine whether oestrogenic endocrine disrupters are disrupting the reproduction of A. rivularis. [ABSTRACT FROM AUTHOR]

• During vitellogenesis GnRHa injections compensate the effects of lower temperature. • fshβ increased in response to HD of GnRH at 27 °C, associated with higher E 2 levels. • GnRHa injected animals had lower vtgA gene expression levels at 20 °C. • GSI and oocyte diameter did not change, but GnRHa compensates fecundity at 20 °C. • Larval production was better in the combined treatment of HD GnRHa and 27 °C. Lambari-do-rabo-amarelo Astyanax altiparanae in the wild reproduce during spring and summer, but females undergo vitellogenesis throughout the year, including the non-spawning winter period when water temperatures are low. The present study investigated the physiological role of temperature modulation on the hypothalamus-pituitary-gonads axis of lambari during winter, as well as the effects of gonadotropin releasing hormone agonist (GnRHa) therapy. Captive females were exposed to two different temperatures (20 °C and 27 °C) and were injected weekly with GnRHa for 21 days during winter (Control, CTR; Low dose; LD and high dose of GnRHa, HD). At the end of the 21-days period gonadosomatic index (GSI), oocyte stage of development and theoretical fecundity were evaluated, together with plasma levels of 17β-estradiol (E 2). Gene expression of the two pituitary gonadotropins follicle-stimulating hormone (fshβ) and luteinizing hormone (lhβ), as well as hepatic vitellogenin-A (vtgA) expression were also analyzed. At the end of the experimental period, females from the six different experimental conditions were induced to spawn using human chorionic gonadotropin (hCG). Spawning performance parameters and plasma levels of the maturation inducing steroid (MIS) were analyzed. Gene expression of fshβ did not change with temperature manipulation, but females exposed to 27 °C and supplemented with a HD of GnRHa exhibited an increased fshβ gene expression, associated with higher E 2 levels. The higher water temperature alone was able to increase E 2 levels. At both water temperatures GnRHa injections induced a decrease in E 2 levels. GnRHa injected females had a lower vtgA gene expression levels at 20 °C. Even with differences in the gene expression of gonadotropins among the various temperature/GnRHa treatments, GSI and oocyte diameter did not change, but GnRHa enhanced the number of vitellogenic oocytes at 20 °C. The reproductive performance of lambari induced to spawn with hCG was better after the combined treatment with GnRHa and summer temperature. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • cGnRH2 increased fshb and lhb mRNA levels in Astyanax altiparanae males. • zGnIH-3 decreased the cGnRH2-induced gonadotropin expression and increased gnrh2 mRNA levels in A. altiparanae males. • zGnIH-3 exerts dual regulatory roles on gonadotropic cells and GnRH2 neurons in A. altiparanae males. Abstract The pituitary gonadotropins, Fsh (follicle-stimulating hormone) and Lh (luteinizing hormone), regulate testicular development and functions in all vertebrates. At the pituitary, different signaling systems regulate the synthesis and secretion of the gonadotropins, such as the hypothalamic neuropeptides GnRH (gonadotropin-releasing hormone) and GnIH (gonadotropin-inhibitory hormone). While GnRH exerts stimulatory roles, the actions of GnIH remain controversial for many teleost species. Therefore, the aim of this study was to evaluate the in vitro effects of chicken GnRH2 (cGnRH2) and zebrafish GnIH-3 (zGnIH-3) on the male gonadotropin and GnRH system expression using pituitary explants and brain slices from a neotropical species with economical and ecological relevance, Astyanax altiparanae. Our results showed that in males, cGnRH2 increased fshb and lhb mRNA levels in the pituitary explants. Interestingly, zGnIH-3 has no effect on basal gonadotropin expression, however zGnIH-3 decreased the cGnRH2-induced fshb and lhb transcripts in male pituitary explants. In the male brain slices, zGnIH-3 showed stimulatory effects, increasing gnrh2 mRNA levels. Overall, our results suggested that GnIH seems to have dual regulatory actions on gonadotropin and GnRH2 expression of A. altiparanae males. This study provided basic information on endocrine regulation of A. altiparanae reproduction, and the obtained results will expand our knowledge, improving the reproductive management of this economically important freshwater species. [ABSTRACT FROM AUTHOR]

Abstract Molecular data of Henneguya chydadea Barassa, Cordeiro and Arana, 2003, found in the gill filaments of Astyanax lacustris bred in fish farm in the State of Mato Grosso do Sul, Brazil was obtained in order to estimate their phylogenetic position among other platysporines myxosporean. The prevalence of the parasite was 28.1% and the range intensity was 1–3 plasmodia per fish. The shape and measurements of mature myxospores were consistent with the characteristics previously defined to H. chydadea. The SSU rDNA sequence of the myxospores of H. chydadea resulted in a total of 1405 nucleotides, and this sequence did not match any of the myxozoan available in the GenBank. Phylogenetic analysis showed H. chydadea within the clade of histozoic myxosporeans and closed together with Henneguya rotunda and Myxobolus pantanalis reported in the gill arch and fins and gill filaments of Salminus brasiliensis respectively. Nonetheless, the SSU rDNA sequences of H. chydadea, H. rotunda and M. pantanalis have only 85.2% and 84.4% similarity, respectively. This is the first molecular study of a Henneguya species that parasitizes a fish belonging to the genus Astyanax in South America. The importance of myxosporeans introduction to new locations along with infected cultured host is emphasized. Graphical abstract Image 1 Highlights • This is the first molecular study of a Henneguya species that parasitizes a South American Astyanax fish. • Phylogenetic position of the pathogenic H. chydadea among other platysporines myxosporeans was determined. • The risk of myxosporeans introduction to new locations along with cultured infected host is emphasized. [ABSTRACT FROM AUTHOR]

Responding appropriately to stress is essential for survival, yet in pathological states, these responses can develop into debilitating conditions such as post-traumatic stress disorder and generalized anxiety. While genetic models have provided insight into the neurochemical and neuroanatomical pathways that underlie stress, little is known about how evolutionary processes and naturally occurring variation contribute to the diverse responses to stressful stimuli observed in the animal kingdom. The Mexican cavefish is a powerful system to address how altered genetic and neuronal systems can give rise to altered behaviors. When introduced into a novel tank, surface fish and cavefish display a stereotypic stress response, characterized by reduced exploratory behavior and increased immobility, akin to "freezing". The stress response in cave and surface forms is reduced by pharmacological treatment with the anxiolytic drug, buspirone, fortifying the notion that behavior in the assay represents a conserved stress state. We find that cave populations display reduced behavioral measures of stress compared to surface conspecifics, including increased time in the top half of the tank and fewer periods of immobility. Further, reduced stress responses are observed in multiple independently derived cavefish populations, suggesting convergence on loss of behavioral stress responses in the novel tank assay. These findings provide evidence of a naturally occurring species with two drastically different forms in which a shift in predator-rich ecology to one with few predators corresponds to a reduction in stress behavior. [ABSTRACT FROM AUTHOR]

Cave-adapted organisms are often characterized by a reduction in pigmentation, eyesight, and enhanced mechanosensory functions. Previous studies have described the genetic basis for a depigmented phenotype in multiple independent populations of the Blind Mexican Tetra, Astyanax mexicanus ; the reduction in melanin content (brown; Mc1r). At least seven wild populations express the brown phenotype. In three populations, there are two different coding sequence alterations affecting Mc1r and the remaining four populations show the accumulation of sequence mutations affecting the 5′ regulatory region. Thus, the Mc1r gene has been the repeated and independent location of mutations in Astyanax . As such, it would appear that this gene is a target during regressive evolution of cave adapted organisms. If this is the case, it would be expected that other cave adapted fish would have mutations in the same gene. We study here the stygobitic catfish Astroblepus pholeter, a depigmented fish found within some river caves in Ecuador. A. pholeter displays mutations in ultra-conserved areas of the pigment-controlling gene, Mc1r, that have been linked to pigment regulation in other organisms. It is thus concluded that Mc1r, a gene known to control pigment variation in many organisms, may be the target of cavernicole regressive evolution across species in different families of fish. [ABSTRACT FROM AUTHOR]

Abstract: We investigated differentiation processes in the Neotropical fish Astyanax that represents a model system for examining adaptation to caves, including regressive evolution. In particular, we analyzed microsatellite and mitochondrial data of seven cave and seven surface populations from Mexico to test whether the evolution of the cave fish represents a case of parallel evolution. Our data revealed that Astyanax invaded northern Mexico across the Trans-Mexican Volcanic Belt at least three times and that populations of all three invasions adapted to subterranean habitats. Significant differentiation was found between the cave and surface populations. We did not observe gene flow between the strongly eye and pigment reduced old cave populations (Sabinos, Tinaja, Pachon) and the surface fish, even when syntopically occurring like in Yerbaniz cave. Little gene flow, if any, was found between cave populations, which are variable in eye and pigmentation (Micos, Chica, Caballo Moro caves), and surface fish. This suggests that the variability is due to their more recent origin rather than to hybridization. Finally, admixture of the young Chica cave fish population with nuclear markers from older cave fish demonstrates that gene flow between populations that independently colonized caves occurs. Thus, all criteria of parallel speciation are fulfilled. Moreover, the microsatellite data provide evidence that two co-occurring groups with small sunken eyes and externally visible eyes, respectively, differentiated within the partly lightened Caballo Moro karst window cave and might represent an example for incipient sympatric speciation. [Copyright &y& Elsevier]

Abstract: Astyanax has become an important model system for evolutionary studies of cave animals. We investigated correlations of population genetic patterns revealed by microsatellite data and phylogeographic patterns shown by mitochondrial DNA sequences in Mexican cave and surface fish of the genus Astyanax (Characidae, Teleostei) to improve the understanding of the colonization history of this neotropical fish in Central and North America and to assess a recent taxonomic classification. The distribution of nuclear genotypes is not congruent with that of the mitochondrial clades. Admixture analyses suggest there has been nuclear gene flow between populations defined by different mitochondrial clades. The microsatellite data indicate that there was mitochondrial capture of a cave population from adjacent populations. Furthermore, gene flow also occurred between populations belonging to different nuclear genotypic clusters. This indicates that neither the nuclear genotypic clusters nor the mitochondrial clades represent independent evolutionary units, although the mitochondrial divergences are high and in a range usually characteristic for different fish species. This conclusion is supported by the presence of morphologically intermediate forms. Our analyses show that the Trans-Mexican Volcanic Belt limited gene flow, but has been crossed by Astyanax several times. In Yucatán, where obvious geographic barriers are missing, the incongruence between the distribution of nuclear and mitochondrial markers reflects random colonization events caused by inundations or marine transgressions resulting in random phylogeographic breaks. Thus, conclusions about the phylogeographic history and even more about the delimitation of species should not be based on single genetic markers. [Copyright &y& Elsevier]

The ability to orient through familiar areas is key to the success of many animal groups. To date, research has focused on how animals orient horizontally and very little work has considered three-dimensional environments, particularly the volumetric surroundings inhabited by many birds, flying insects and fish. These animals do not have their movement constrained by surfaces, and can move with three degrees of freedom. This extra freedom of movement increases the amount of navigational information potentially available, which could lead to an information-processing problem. We studied how a fish, the sighted banded tetra, Astyanax fasciatus, copes with this problem by testing how it learns and stores information from its volumetric surroundings. Using a novel assay based on associative learning of the vertical (up/down) and horizontal (left/right) components of a rotating Y-maze, we found that banded tetras learned and remembered information from the vertical and horizontal components when they were presented either separately or as an integrated three-dimensional unit. Furthermore, when information from the two components conflicted, the fish used the previously learned vertical information in preference to the horizontal, possibly because the vertical axis contained an extra, global, cue: hydrostatic pressure. We propose that animals that navigate through volumes may simplify the information storage problem by encoding the horizontal and vertical components separately. These components could then be used together for rapid, efficient orientation. [Copyright &y& Elsevier]

Abstract: Fish (Astyanax sp.) were exposed to water samples from the Arroio Saldanha, locality of an oil spill, and the Barigui River (upstream and downstream of the confluence of the river and Arroio Saldanha), Brazil. Histopathological and biochemical biomarkers, as acetylcholinesterase (AchE), ethoxyresorufin-O-deethylase (EROD), catalase (CAT), glutathione-S-transferase (GST) activities, lipid peroxidation (LPO) levels, and amounts of polynuclear aromatic hydrocarbon residues (PAHs) in bile were evaluated. PAHs, metals, and surfactants were investigated in the water samples. CAT and GST activities increased in the Arroio Saldanha sample, whereas CAT activity and LPO levels were increased in the upstream and downstream sites. The results suggested a toxic action of the free radicals and disturbance of the antioxidant defense mechanisms. However, the EROD and AchE activities were not affected. Histopathology studies showed severe lesions. The oil is still bioavailable to biota, moreover, other pollution sources continue to affect the water in the Barigui River. [Copyright &y& Elsevier]

Abstract: The lens influences retinal growth and differentiation during vertebrate eye development but the mechanisms are not understood. The role of the lens in retinal growth and development was studied in the teleost Astyanax mexicanus, which has eyed surface-dwelling (surface fish) and blind cave-dwelling (cavefish) forms. A lens and laminated retina initially develop in cavefish embryos, but the lens dies by apoptosis. The cavefish retina is subsequently disorganized, apoptotic cells appear, the photoreceptor layer degenerates, and retinal growth is arrested. We show here by PCNA, BrdU, and TUNEL labeling that cell proliferation continues in the adult cavefish retina but the newly born cells are removed by apoptosis. Surface fish to cavefish lens transplantation, which restores retinal growth and rod cell differentiation, abolished apoptosis in the retina but not in the RPE. Surface fish lens deletion did not cause apoptosis in the surface fish retina or affect RPE differentiation. Neither lens transplantation in cavefish nor lens deletion in surface fish affected retinal cell proliferation. We conclude that the lens acts in concert with another optic component, possibly the RPE, to promote retinal cell survival. Accordingly, deficiency in both optic structures may lead to eye degeneration in cavefish. [Copyright &y& Elsevier]

Abstract: The relative condition factor (Kn), gonadosomatic index (GSI), selected hematological variables and gill morphology of the fish Astyanax fasciatus were analyzed in two sites (site 1 was unpolluted and site 2 was polluted with untreated domestic sewage) of a tropical river (Camanducaia river, São Paulo State, Brazil). The relationship between the body mass (MB) and the standard length (LS) of A. fasciatus from both sites was MB=0.00799 LS3.51843. The Kn values from both males and females and the GSI of females were significantly higher in site 2. The mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were higher in females from site 2. Gill tissue anomalies and gill parasites were rare in fish from both sites; however, the number of chloride cells was significantly higher in fish from site 1. A. fasciatus presents high capacity to live in ion-poor and soft water and is able to compensate for environmental changes caused by untreated domestic sewage discharges. [Copyright &y& Elsevier]

Although much is now known about the mechanisms that insects, birds and mammals use to orient within familiar areas, our knowledge of such mechanisms in fish is scant. I used the transformational approach to test whether the blind Mexican cave fish can encode shape and size in an internal representation of space. These fish are excellent study animals, as they swim at high velocities (presumably to enhance lateral line organ stimulation) when faced with unfamiliar landmarks or environments. As they are blind, potentially confounding cues from visual global landmarks are unavailable. The fish learnt a square configuration of four landmarks and so must have been be able to encode spatial relationships between the elements within this configuration. After learning landmark arrays, the cave fish showed significant dishabituation (swimming velocity was increased) when exposed to landmark transformations. The fish must therefore have been comparing the environment that they perceived with an internal representation of the environment that they had learnt. The results show that blind Mexican cave fish can encode size (absolute distance between landmarks) and possibly also shape within their spatial maps. [Copyright &y& Elsevier]

The present study describes the morphological, histopathological and SSU rDNA data of a new myxosporean species infecting farmed Astyanax lacustris fish from the state of São Paulo, Brazil. Henneguya lambariensis sp. nov. was found infecting the gills, and the plasmodial development resulted in displacement, blood congestion, compression, deformation and lamellar fusion, stretching of the epithelia, hyperplasia of the epithelial cells, edema, and mild infiltration of the mast cells and lymphocytes. The SSU rDNA sequencing resulted in the sequencing of 1804 nucleotides that did not correspond to any myxosporean sequences deposited in GenBank. The closest phylogenetic affinity of the new species was to the South American Henneguya loreotoensis and Henneguya guanduensis, which also parasite the gills. The present study suggests this new myxosporean species has considerable pathogenic potential, and health monitoring should be implemented in A. lacustris fish farms to ensure production. • Taxonomic descriptions of a myxosporean species infecting an economic important fish species from fish farms in Brazil. • Morphological, histological and molecular analysis identified a new Henneguya species in freshwater fish. • The new Henneguya species has considerable pathogenic potential. [ABSTRACT FROM AUTHOR]

Atrazine (ATZ) is among the most widely used herbicides in the world, and yet it has a potential to contaminate aquatic environments due to pesticide leaching from agricultural areas. In the Neotropical region, studies about the effects of this herbicide in native aquatic wildlife is scarce.Our study aimed at investigating the effects of a 30-day exposure to a commercial atrazine formulation on oxidative stress parameters, histopathology in testis and liver, and hormone levels in males and female of yellow-tailed tetra fish (Astyanax altiparanae). Adults were exposed to low but environmentally relevant concentrations of atrazine as follows: 0 (CTL-control), 0.5 (ATZ0.5), 1 (ATZ1), 2 (ATZ2) and 10 (ATZ10) μg/L. Our results showed decreased GST activity in gills in all groups of exposed animals and increased CAT activity in gills from the ATZ10 group. In the liver, there was an increase in lipid peroxidation in fish from ATZ1 and ATZ2 groups. Histological analysis of the liver showed increased percentage of sinusoid capillaries in ATZ2 fish, increased vascular congestion in ATZ1 and increased leukocyte infiltration in the ATZ10 group. Hepatocyte diameter analysis revealed a decrease in cell size in all groups exposed to ATZ, and a decrease in hepatocyte nucleus diameter in ATZ1, ATZ2 and ATZ10 groups. Endocrine parameters did not show significant changes following ATZ exposure, although an increase of triiodothyronine/thyroxine (T 3 /T 4) ratio was observed in ATZ2 fish. Our results provide evidence that even low, environmentally relevant concentrations of ATZ produced oxidative damage and histological alterations in adult yellow-tailed tetra. • Exposure to low atrazine concentrations induced oxidative stress in gills, liver and muscles in yellow-tailed tetra fish. • Atrazine induced histological damage in the liver and testis. • Atrazine induced little endocrine disruption, mainly altering triiodothyronine/thyroxine (T3/T4) ratio. [ABSTRACT FROM AUTHOR]

The reproductive physiology of fish can be changed by the presence of pollutants in the water, which act as endocrine disrupting compounds (EDC). We evaluated the impacts of water contaminants in polluted reservoirs acting as possible EDC on the reproductive physiology of Astyanax fasciatus and Hoplias malabaricus males. We used biomarkers with different levels of biological organization. Hoplias malabaricus adult males were collected in the summer and winter at five different sites in the Tietê River Basin: the Ponte Nova reservoir (PN), considered a reference site due to the low anthropogenic influence; the Billings reservoir (BIL) at two different branches; and the Guarapiranga reservoir (GUA) at two different branches. Astyanax fasciatus adult males were collected at PN and BIL. BIL and GUA are subjected to great anthropogenic action. We analyzed gonadal histomorphology, testosterone (T), 11-ketotestosterone (11-KT), estradiol (E 2) plasma levels, and gene expression of hepatic vitellogenin (vtgA) and pituitary follicle stimulating hormone (fshβ). In the PN reservoir (reference), the biomarkers analyzed in both species did not differ between the periods analyzed. This is an evidence that the animals keep the same reproductive activity during both seasons. The changes in the plasma concentration of gonadal steroids in both species in polluted reservoirs suggest the presence of EDC compounds in the water and/or adjusts of the physiological setpoint to allow the reproduction in such adverse conditions. The use of vtgA as biomarker suggests the presence of estrogenic compounds, mainly in BIL, but with a more evident response of H. malabaricus. However, even considering physiological changes, both species present testes during the maturation phase that allow the reproduction in an environment with a high degree of pollution. • Steroid data suggest a response to EDC and/or adjusts to reproduce in polluted water. • vtgA expression in males suggests the presence of estrogenic compounds in reservoirs. • The estrogenic response is more evident in Hoplias malabaricus. • Summer is the season with the main responses of the biomarkers. • Both species present mature testes allowing reproduction in polluted reservoirs. [ABSTRACT FROM AUTHOR]

Brazilian freshwater ecosystems are continuously exposed to pesticides and domestic sewage. The Uruguay River was chosen for this study because of its international importance, as it flows through Brazil, Argentina, and Uruguay. It receives contaminants such as pesticides and domestic residues. Thus, the aim of this study to assess the accumulation of pesticides in muscle of the fish Astyanax jacuhiensis , its biochemical responses, and the presence of pesticides in water. In total, seven pesticides were registered in water from both river sites. Eight pesticides were detected in fish muscle. The biochemical responses showed that brain lipid peroxidation (LPO) and protein carbonyl (PC) in A. jacuhiensis were higher in the summer. Muscle showed the highest LPO levels in the spring and the highest PC in the summer. Liver LPO and PC levels were higher in the spring and summer. In the gills, the PC was higher in the spring and the LPO in the spring and winter. In the brain and in the gills, glutathione-S-transferase activity was high in the summer and autumn. Catalase activity was lower during the winter and spring. Non-protein thiol (NPSH) levels were lower in the brain in the winter and spring. Muscle tissue showed lower NPSH in the winter (site 1). Liver NPSH showed increased levels in liver in the spring and winter (site 2). The biochemical results clearly is related to pesticides and/or to the presence of other contaminants in the water such as metals or domestic sewage. The accumulation of pesticides in fish muscle added evidence that pesticides have been used in the area surrounding the Uruguay River. In conclusion, the biomarkers assayed in the present study could be used in future investigations considering other sampling sites along Uruguay River. Image 1 • The results highlight the contamination of the Uruguay River by pesticides. • Muscle of Astyanax jacuhiensis showed a diversity of pesticides. • Fish tissues exhibited oxidative damage due to pesticide presence in the muscle. • Summer showed highest concentration of different pesticides in fish muscle. [ABSTRACT FROM AUTHOR]

Bioindicator organisms are important tools in environmental monitoring studies. Understanding this, the overall goal of the present study was to evaluate the sensitivity and viability of the native fish species Banded tetra, (Astyanax aeneus ; Günther, 1860), widely spread in the aquatic ecosystems of the Yucatan Peninsula in Mexico, as a bioindicator organism. In order to do this, we performed a bioassay at sublethal concentrations using copper (CuSO 4) to experimentally evaluate and validate the relationship between the trace metals and oxidative stress biomarkers response [(catalase (CAT), lipoperoxidation content (LPO)], detoxification [(glutathione S-transferase (GST), metallothionein content (MT)] and neurotoxicity (AChE) in muscle of A. aeneus. Results showed changes in biomarkers after 96 h: Catalase activity (CAT) was significantly higher above 1.5 and 2 mg/L (154.35 and 172.50% increase, respectively); lipid peroxidation contents (TBARS), GST activity, and MT content were very similar to CAT activity at 1.5 and 2 mg/L of Cu. In terms of neurotoxicity, AChE activity was significantly inhibited at 0.1 mg/L (64%; p < 0.001) and 0.5 mg/L (44%; p < 0.001) of Cu. Based on the bioassay results, we performed a trace metal monitoring campaign in muscle of A. aeneus caught in 15 sites with different anthropogenic activities, during the summer of 2017, to establish a baseline of trace metals pollution in the state of Campeche. A. aeneus showed the highest trace metal accumulation in the following order: Al > Fe > Mn > Zn > Cu > Hg > Cr > Pb > Cd > V > As, while sites were arrange as follows: Xnoha lagoon > Palizada River > Candelaria River > Ululmal > Maravillas > López Mateos. PCA showed a cluster between biomarkers (GST, CAT, TBARS, and MT) and concentration of metals (Cd, Cu, Fe, Zn, Hg and Cr). Conversely, AChE inhibition was not related to a specific metal, but highest inhibitions (>50%) were present in those sites with intensive agricultural practices. These results determined that, based on its physiological response and trace metal bioaccumulation, Astyanax aeneus can be considered a good bioindicator for evaluating the presence of trace metals in tropical aquatic systems of the Yucatan Peninsula. Image 1 • Astyanax aeneus was chosen for the determination of trace metals in aquatic ecosystems. • Trace metals and biomarkers were analyzed and validate in wild Astyanax aeneus muscle. • Oxidative stress biomarkers are sensitive in this native fish to trace metal pollution. • Astyanax aeneus is a good bioindicators to metal exposure in aquatic ecosystems. [ABSTRACT FROM AUTHOR]

Increased malondialdehyde (MDA) levels are commonly considered an indicator of lipid peroxidation derived from oxidative stress insults promoted by exposure of fish to pollutants. However, a decrease in MDA levels after xenobiotic exposure has been also reported, an effect that is mostly attributed to enhanced antioxidant defenses. In this study, we assessed whether pollutant-mediated MDA decrease would be associated with antioxidant enhancement or with its metabolism by aldehyde dehydrogenase (ALDH) in the liver and gills of lambari (Astyanax altiparanae) exposed to diesel oil (0.001, 0.01, and 0.1 mL/L). MDA levels were decreased in the liver of lambari exposed to diesel. The activities of the antioxidant enzymes, catalase (CAT) and glutathione peroxidase (GPx), were unchanged in the liver, while that of glucose-6-phosphate dehydrogenase (G6PDH) was decreased. In contrast, levels of total glutathione (tGSH) and the activity of glutathione S -transferase (GST) were increased in the liver, which partly support antioxidant protection against lipid peroxidation. More importantly, ALDH activity increased in a concentration-dependent manner, being negatively correlated with MDA levels, indicating MDA metabolism by ALDH. In the gills, diesel exposure increased MDA and lipid hydroperoxide levels, and promoted increases in antioxidant defenses, indicating oxidative stress. Curiously, ALDH activity was undetectable in the gills, supporting the possibility of direct MDA excretion in the water by the gills. Analyses of MDA in the water revealed increased levels of MDA in the aquaria in which the fish were exposed to diesel, compared to control aquaria. A second experiment was carried out in which the fish were intraperitoneally injected with MDA (10 mg/kg) and analyzed after 1, 6, and 12 h. MDA injection caused a time-dependent decrease in hepatic MDA levels, did not alter ALDH, CAT, GPx, and GST activities, and decreased G6PDH activity and tGSH levels. In the gills, MDA injection caused a slight increase in MDA levels after 1 h, but did not alter GPx, G6PDH, and GST activities. MDA injection also enhanced CAT activity and tGSH levels in the gills. MDA concentration in water increased progressively after 1, 6, and 12 h, supporting the hypothesis of direct MDA excretion as an alternative route for MDA elimination in fish. Our results suggest that the decreased MDA levels after exposure of lambari to diesel oil pollutant probably reflects an association between enhanced antioxidant protection, MDA metabolism, and MDA excretion in water. • Diesel exposure decreased MDA levels in fish liver. • MDA decrease in the liver was associated with increased ALDH activity. • ALDH was undetectable in gills. • Diesel exposure and MDA injection resulted in increased MDA levels in water. • MDA decrease seems to be related to antioxidant increase, mda metabolism and excretion. [ABSTRACT FROM AUTHOR]