A biomarker approach to study the effects of polluted Brazilian urban reservoirs in a native fish.

Two of the largest water reservoirs in the Metropolitan Region of São Paulo, Brazil (MRSP), named Billings and Guarapiranga, are facing high levels of anthropic impact. This is evidenced by the presence of contaminants and pollutants, which are deteriorating their water quality. Therefore, this study evaluated antioxidant defense enzymes, lipoperoxidation and genotoxicity, in adult females of a native species, Astyanax altiparanae from the Guarapiranga and Billings reservoirs. The study also aimed to evaluate these biomarkers during two different periods of the year, the rainy (summer) and dry (winter) seasons. The oxidative stress was evaluated by the activity of enzymes such as glutathione peroxidase, glutathione S-transferases, superoxide dismutase, and catalase in the gills and liver, and the occurrence of lipoperoxidation was also evaluated in both organs. The genotoxicity was assessed by performing comet assay, micronucleus, and nuclear abnormality tests on blood samples. The results showed that fish from both reservoirs are subjected to oxidative stress and genotoxic damage, mainly during winter, but fish living in Billings showed greater alterations than fish from Guarapiranga. Likewise, the results of the principal component analysis suggested that caffeine, nitrogenous compounds, and some metals might be triggering these toxic effects in fish.

Early ontogeny of the freshwater fish Rhytiodus microlepis (Characiformes, Anostomidae) from the Amazon basin.

The early development of the freshwater fish Rhytiodus microlepis is characterized by the description of external morphological, meristic, and morphometric changes, as well as the growth patterns, thereby establishing a reference for the identification of its larvae and juveniles. Specimens were collected from the Amazon river channel and floodplain. Ninety-seven individuals were analysed with standard length varying between 4.31 and 79.23 mm. Rhytiodus microlepis larvae are altricial, with an elongated and fusiform body, anal opening reaching the middle region of the body, and simple nostrils becoming double and tubular during development. The pigments vary from one to two chromatophores in the dorsal region of the head in pre-flexion and flexion, but later the pigmentation pattern intensifies, transverse bands appear along the body, and a conspicuous spot appears in the basal region of the caudal fin. The total number of myomeres ranges from 49 to 50. During the transition from larval (post-flexion) to the juvenile periods, the most significant anatomical changes occur, such as the presence of all fins and increased body pigmentation. Integrated myomere count and pigmentation pattern are effective for the correct identification of the initial life stages of R. microlepis from the Amazon basin. Our results expand the knowledge about the early life history of Neotropical freshwater fish species.

How does temperature rise affect a freshwater catfish Rhamdia quelen? A proteomic approach.

Outside of scientific circles, climate change is a hotly debated topic due to all its consequences. Changes in the temperature can affect aquatic organisms and it is important to understand and to detect earlier signals. This study aimed to analyze how a Neotropical fish species responds to temperature increases, using proteomic analysis as a tool. For this, fish of the species Rhamdia quelen, male and female, were exposed to two temperatures: 25 °C and 30 °C. After 96 h, the animals were anesthetized, euthanized and the liver was collected for proteomic analysis. Using freely available online software and databases (e.g. MetaboAnalyst, Gene Ontology and UniProt), we define the altered proteins in both sexes: 42 in females and 62 in males. Data are available via ProteomeXchange with identifier PXD046475. Differences between the two temperatures were observed mainly in the amino acid metabolic pathways. The cellular process and the immune response was altered, indicating that effects at lower levels of biological organization could serve as a predictor of higher-level effects when temperature rise affects wildlife populations. Thus, we conclude that the increase in temperature is capable of altering important cellular and physiological processes in R. quelen fish, with this response being different for males and females.

On the lectotype designation of Bryconamericus rubropictus (Berg) (Characiformes, Characidae, Stevardiinae).

The lectotype specimen of Bryconamericus rubropictus (Berg) and its designation have remained imprecisely documented since its publication. The lack of a photograph or an unambiguous illustration, correct size, inaccurate labelling, and proper specimen separation has led to an uncertainty about the identity and nomenclatural status of the lectotype. We recovered and provided detailed morphological data on the specimen that must be recognised as the lectotype. This contribution brings stability and clarity on the nomenclatural status of the species and its type material.

Experimentally exposed toxic effects of long-term exposure to environmentally relevant concentrations of CIP in males and females of the silver catfish Rhamdia quelen.

Ciprofloxacin (CIP) is an antibiotic commonly used in human and veterinary medicine. It is present in the aquatic environment, but we still know very little about its effect on non-targeted organisms. This study aimed to evaluate the effects of long-term exposure to environmental CIP concentrations (1, 10, and 100 µg.L-1) in males and females of Rhamdia quelen. After 28 days of exposure, we collected the blood for the analysis of hematological and genotoxic biomarkers. Additionally, we measured 17 ß-estradiol and 11 keto-testosterone levels. After the euthanasia, we collected the brain and the hypothalamus to analyze acetylcholinesterase (AChE) activity and neurotransmitters, respectively. The liver and gonads were assessed for biochemical, genotoxic, and histopathological biomarkers. At 100 µg.L-1 CIP, we observed genotoxicity in the blood, nuclear morphological changes, apoptosis, leukopenia, and a reduction of AChE in the brain. In the liver was observed oxidative stress and apoptosis. At 10 µg.L-1 CIP, leukopenia, morphological changes, and apoptosis were presented in the blood and a reduction of AChE in the brain. Apoptosis, leukocyte infiltration, steatosis, and necrosis occurred in the liver. Even at the lowest concentration (1 µg.L-1), adverse effects such as erythrocyte and liver genotoxicity, hepatocyte apoptosis, oxidative stress, and a decrease in somatic indexes were observed. The results showed the importance of monitoring CIP concentrations in the aquatic environment that cause sublethal effects on fish.

Un caso de leucismo en el Juil de Jamapa, Rhamdia laticauda (Siluriformes, Heptapteridae) en el noreste del estado de Oaxaca, México

Se reporta la primera observación de un espécimen leucístico en el Juil de Jamapa, Rhamdia laticauda en el noreste del estado de Oaxaca. En un estudio ictiológico de la Reserva de la Biosfera Tehuacán-Cuicatlán en el río Xiquila de la cuenca alta del río Papaloapan, el pez se capturó con pesca eléctrica y se fotografió. El bagre vivo mostró una reducción de pigmentos en la piel, manchas blancas y ojos negros normales. Esto contribuye al conocimiento de la variabilidad intraespecífica y la historia natural de R. laticauda. Se discuten las causas potenciales de esta anomalía de color.

Here, we report the first observation of a leucistic specimen in the Rock catfish, Rhamdia laticauda in the Northeast of Oaxaca, Mexico. We caught the specimen by electric fishing and then photographed during an ichthyological survey carried out in the Tehuacán-Cuicatlán Biosphere Reserve in the Xiquila River of the upper Papaloapan River basin. The live catfish showed a lack of pigment in the skin, white spots, and normal eye color. This information contributes to the knowledge about intraspecific variability and natural history of R. laticauda. We discuss the potential causes of this color anomaly.

Integrative study of the reproductive biology and growth of Acestrorhynchus pantaneiro Menezes, 1992 (Characiformes, Acestrorhynchidae).

We studied the reproductive strategy, sexual system and growth of dientudo paraguayo Acestrorhynchus pantaneiro. After 2 years of monitoring in shallow areas of a floodplain lake from the lower Paraná basin (Argentina), it was evidenced that water temperature modulated gonadal maturation, but it was the river water level the synchronising stimulus that triggered spawning. This species exhibited a single annual breeding period from October to January, with most spawning activity in November. According to the von Bertalanffy growth curve, fish would reach autumn to winter months with LS of ~120 mm, already mature males. The first mature females were found at LS of 210 mm, becoming sexually mature between the second and third breeding seasons. This is the first integrative study that includes the body-length frequency distribution and sex differential size at first maturity and growth, and reports the presence of intersex gonads questioning its sexual pattern from gonochoristic to sequential hermaphrodite species. The sexual pattern, the multiple spawning behaviour and a medium to high absolute fecundity support the opportunistic and invasive behavior observed in previous contributions for this characiform species.

Chromosomal Mapping of the Histone Multigene Family and U2 snRNA in Hypancistrus Species (Siluriformes, Loricariidae) from the Brazilian Amazon.

Loricariidae (Siluriformes) comprises ∼1026 species of neotropical fish, being considered the most diverse among the Siluriformes. Studies on repetitive DNA sequences have provided important data on the evolution of the genomes of members of this family, especially of the Hypostominae subfamily. In this study, the chromosomal mapping of the histone multigene family and U2 snRNA was performed in two species belonging to the Hypancistrus genus, Hypancistrus sp. "pão" (2n = 52, 22m + 18sm +12st) and Hypancistrus zebra (2n = 52, 16m + 20sm +16st). The presence of dispersed signals of histones H2A, H2B, H3, and H4 in the karyotype of both species, with each sequence displaying a varied level of accumulation and dispersion of these sequences between them was observed; in addition, U2 snDNA probe only showed positive results in H. zebra, which present this multigene in the terminal region of three chromosomal pairs. The obtained results resemble data already analyzed in the literature, in which the action of transposable elements interfere in the organization of these multigene families, in addition to other evolutionary processes that shape the evolution of the genome, such as circular or ectopic recombination. This study also shows that the dispersion of the multigene histone family is quite complex, and from this, these data serve as a point of discussion for the evolutionary processes that occur in the Hypancistrus karyotype.

Effects of live prey concentration, salinity, and weaning age on larviculture of Piaractus brachypomus reared in a recirculating aquaculture system.

This study evaluated the effects of live prey concentration (nauplii of Artemia sp.), water salinity, and weaning age on survival, growth, and stress resistance rate (Rs) of Piaractus brachypomus under larviculture in a recirculating aquaculture system (RAS). Larvae aged 6 days post-hatching (1.64 ± 0.11 mg) were distributed in 28-L tanks (five larvae L-1), in two RASs. The experiment was carried in a 2 × 2 × 2 factorial arrangement, as follows: two feeding strategies (sudden transition from live food to commercial food after 10 (FT10) and 20 (FT20) days of larviculture with Artemia); two daily initial prey concentrations (P350 = 350 and P700 = 700 nauplii larva-1, these being increased every 5 days); and two water salinities (S0 = fresh water and S2 = 2 g of salt L-1). Weight (W), total length (TL), and daily specific growth rate (SGR) were evaluated after 10, 20, 30, and 40 days of larviculture. After 40 days of larviculture, survival was evaluated and a test of air exposure was performed to determine stress resistance rate (Rs). Noteworthy results during this period are the lowest specific daily growth rate (SGR) after weaning for FT10 and the best growth results for S2 and P700. After 40 days, weight (W) and total length (TL) showed effects of P, FT, and S with higher values for P700, FT20, and S2 (P < 0.05). The interaction P × FT × S also had effects on survival and Rs at the end of the experiment, with higher survival and Rs for P700FT20S2 (P < 0.05). Larviculture of P. brachypomus in RAS, in association with the three managements-live prey concentration P700, salinity S2, and age at feed transition FT20-promotes maximization of survival, growth, and stress resistance rate of the animals. The larviculture of P. brachypomus in RAS must be carried out with an initial concentration of live prey of 700 nauplii larva-1, at a salinity of 2 g of salt L-1 and with the feeding transition starting in 20 days of larviculture, for maximization intensive larviculture of this species.

Early ontogeny of the commercially valuable fish red-bellied pacu Piaractus brachypomus (Characiformes, Serrasalmidae) from the Amazon, Brazil.

The initial development of the red-bellied pacu Piaractus brachypomus is described using morphological, meristic and morphometric characteristics. A total of 127 individuals were analysed (47 in the yolk-sac, 35 in pre-flexion, five in flexion, 20 in post-flexion and 20 in juvenile) with standard length varying between 2.92 and 48.61 mm. The larvae are born poorly developed and have a discoidal yolk at ~6.33 mm standard length. During early ontogeny, the mouth passes from terminal to subterminal and the anal opening reaches the vertical line over the midline region of the body. There are changes in body shape from long and moderate to deep, head length from small to large, and eye diameter from moderate to large. Dendritic chromatophores were present in the ventral, dorsal and upper part of the swim bladder in the early larval stages. Rounded spots are evident all over the body in juveniles. The total number of myomeres ranges from 39 to 41 (20-23 pre-anal, 17-20 post-anal). Through the morphometric relationships, it was evidenced that the greatest changes during the initial ontogeny of P. brachypomus occur in the transition from the post-flexion stage to the juvenile period, indicating changes in behaviour, foraging and physiology.

Hatchery tanks induce intense reduction in microbiota diversity associated with gills and guts of two endemic species of the São Francisco River.

The São Francisco River (SFR), one of the main Brazilian rivers, has suffered cumulative anthropogenic impacts, leading to ever-decreasing fish stocks and environmental, economic, and social consequences. Rhinelepis aspera and Prochilodus argenteus are medium-sized, bottom-feeding, and rheophilic fishes from the SFR that suffer from these actions. Both species are targeted for spawning and restocking operations due to their relevance in artisanal fisheries, commercial activities, and conservation concerns. Using high-throughput sequencing of the 16S rRNA gene, we characterized the microbiome present in the gills and guts of these species recruited from an impacted SFR region and hatchery tanks (HT). Our results showed that bacterial diversity from the gill and gut at the genera level in both fish species from HT is 87% smaller than in species from the SFR. Furthermore, only 15 and 29% of bacterial genera are shared between gills and guts in R. aspera and P. argenteus from SFR, respectively, showing an intimate relationship between functional differences in organs. In both species from SFR, pathogenic, xenobiont-degrading, and cyanotoxin-producer bacterial genera were found, indicating the critical pollution scenario in which the river finds itself. This study allowed us to conclude that the conditions imposed on fish in the HT act as important modulators of microbial diversity in the analyzed tissues. It also raises questions regarding the effects of these conditions on hatchery spawn fish and their suitability for restocking activities, aggravated by the narrow genetic diversity associated with such freshwater systems.

Nutritional feed additives reduce the adverse effects of transport stress in the immune system of Tambaqui (Colossoma macropomum).

Here we show that selected nutritional feed additives reduce the adverse effects of transport stress on the immune system and hematology of tambaqui. We formulated a control diet to contain normal dietary levels of vitamin E (21.6 mg/ kg diet) and C (143 mg/kg), then we added supra levels of these vitamins (vit E - 264 mg/kg and vit C - 1000 mg/kg) to a second diet. Finally, a third diet was produced to contain similar levels of vitamins from diet 2 with 0.1% beta-glucan supplementation. Four hundred thirty-two tambaquis (20.91 g ± 0.27 g) were randomly assigned to 12 aquaria and fed the diets for 15 days; then, all fish were transported for five h and then returned to the aquaria. Blood samples were collected before and after the transport and at the end of the trial (60 days). Transportation significantly increased blood glucose that returned to baseline levels at the end of the trial. However, cortisol seemed to be unresponsive to the stress. Surprisingly, the stress significantly increased the immunoglobulin level after transport. Additionally, the transport markedly reduced the red blood cell count and leukocyte and lymphocytes counts while increasing the control group's neutrophil number. These effects lasted until the end of the trial in the control group. Supra levels of the vitamins and glucan supplementation prevented the decrease in red blood cell and leukocyte count after the stress. Additionally, beta-glucan supplementation induced lower cortisol levels in all the sampling points. However, the effect on the immune parameters was limited, increasing only the lysozyme activity and serum protein levels in the beta-glucan supplemented group and the group fed only the supra levels of vitamins, respectively. In sum, our results indicated that transport for five h induced a limited effect on stress biomarkers. The use of supra levels of antioxidant vitamins alone or in combination with beta-glucan could restore or prevent the adverse effects of stress on hematology and the immune system.

Phylogenomic and expression analysis of Colossoma macropomum cyp19a1a and cyp19a1b and their non-classical role in tambaqui sex differentiation.

The genes coding for Cytochrome P450 aromatase (cyp19a1a and cyp19a1b) and estrogen (E2) receptors (esr1, esr2a and esr2b) play a conserved role in ovarian differentiation and development among teleosts. Classically, the "gonad form" of aromatase, coded by the cyp19a1a, is responsible for the ovarian differentiation in genetic females via ligation and activation of the Esr, which mediates the endocrine and exocrine signaling to allow or block the establishment of the feminine phenotype. However, in neotropical species, studies on the molecular and endocrine processes involved in gonad differentiation as well as on the effects of sex modulators are recent and scarce. In this study, we combined in silico analysis, real-time quantitative PCR (qPCR) assay and quantification of E2 plasma levels of differentiating tambaqui (Colossoma macropomum) to unveil the roles of the paralogs cypa19a1a and cyp19a1b during sex differentiation. Although the synteny of each gene is very conserved among characids, the genomic environment displays striking differences in comparison to model teleost species, with many rearrangements in cyp19a1a and cyp19a1b adjacencies and transposable element traces in both regulatory regions. The high dissimilarity (DI) of SF-1 binding motifs in cyp19a1a (DI = 10.06 to 14.90 %) and cyp19a1b (DI = 8.41 to 13.50 %) regulatory region, respectively, may reflect in an alternative pathway in tambaqui. Indeed, while low transcription of cyp19a1a was detected prior to sex differentiation, the expression of cyp19a1b and esr2a presented a large variation at this phase, which could be associated with sex-specific differential expression. Histological analysis revealed that anti-estradiol treatments did not affect gonadal sex ratios, although Fadrozole (50 mg kg-1 of food) reduced E2 plasma levels (p < 0,005) as well cyp19a1a transcription; and tamoxifen (200 mg kg-1 of food) down regulated both cyp19a1a and cyp19a1b but did not influence E2 levels. Altogether, our results bring into light new insights about the evolutionary fate of cyp19a1 paralogs in neotropical fish, which may have generated uncommon roles for the gonadal and brain forms of cyp19a1 genes and the unexpected lack of effect of endocrine disruptors on tambaqui sexual differentiation.

Regulación genética de la determinación sexual y diferenciación gonadal en peces teleósteos / Genetic regulation of sex determination and gonadal differentiation in teleost fish / Regulação genética da determinação sexual e diferenciação gonadal em peixes teleósteos

RESUMEN Comprender el control de la determinación del sexo y la diferenciación sexual en peces es fundamental para mejorar aspectos de manejo, productividad, economía y conservación de las especies. El objetivo de esta revisión es brindar información de los principales mecanismos genético-moleculares de determinación y diferenciación sexual en peces teleósteos. La búsqueda de información se desarrolló entre 2019 - 2021 a través de bases de datos bibliográficas utilizando frases como: "sex determination fish", "sexual differentiation fish'" y "sex neotropical fish". La selección de la información se realizó llevando en consideración máximo 10 años de publicación, descartando documentos considerados como tesis de maestra o doctorado. La determinación del sexo puede ser definido por sistemas cromosómicos como XX/XY ZZ/ZW XX/X0, ZZ/Z0, XXI, XX2 y XIX2Y o modulado por diferentes genes autosómicos tales como cyp19al , fox12, figla, dmrtl , sox9, amh ygsdf sin embargo, a pesar de los grandes avances en la investigación en el área molecular; el proceso de regulación en la determinación y diferenciación del sexo en peces aún no está completamente dilucidado, especialmente en especies Neotropicales.

AВSTRАСT Understanding the control of sex determination and sexual differentiation in fish is essential to improve aspects of management, productivity economy and conservation of the species. The objective of this review is to provide information on the main genetic-molecular mechanisms of sexual determination and differentiation in teleost fish. The information search was developed between 2019 - 2021 through bibliographic databases using phrases such as: "sex determination fish", "sexual differentiation fish" and "sex neotropical fish". The selection of the information was carried out taking into consideration a maximum of 10 years of publication, discarding documents considered as master's or doctoral theses. The sex determination can be defined by chromosome systems such as XX/XY, ZZ/ZW, XX/X0, ZZ/ Z0, XXI, XX2 and XIX2Y or modulated by different autosomal genes such as cyp19al1 fox12, figla, dmrtl: sox9, amh, and gsdf, However despite the great advances in research in the molecular area, the regulation process in the determination and differentiation of sex in fish is not yet fully elucidated, especially in species Neotropical.

RESUMO Compreender o controle da determinação do sexo e a diferenciação sexual nos peixes é fundamental para melhorar a gestão, produtividade, economia e conservação das espécies. O objetivo desta revisão é fornecer informação sobre os principais mecanismos genéticos-moleculares da determinação e diferenciação sexual em peixes teleósteos. A busca da informação foi realizada entre 2019 - 2021 através de bases de dados bibliográficas, utilizando frases como: "sex determination fish", ""sexual differentiation fish" y ""sex neotropical fish". A seleção da informação foi realizada levando em consideração no máximo 10 años de publicação, descartando-se documentos considerados como teses de mestrado ou doutorado. A determinação do sexo pode ser definida por sistemas cromossômicos como XX/XY, ZZ/ZW XX/X0, ZZ/Z0, XXl, XX2 e XIX2Y ou modulada por diferentes genes autossômicos tais como cyp19al , fox12, figla, dmrtl1 sox9, amh e gsdf no entanto, apesar dos grandes avanços na pesquisa molecular o processo de regulação na determinação e diferenciação do sexo nos peixes ainda não está totalmente elucidado, especialmente nas espécies Neotropicais.

Genomic selection signatures in farmed Colossoma macropomum from tropical and subtropical regions in South America.

Tambaqui or cachama (Colossoma macropomum) is one of the most important neotropical freshwater fish used for aquaculture in South America, and its production is concentrated at low latitudes (close to the Equator, 0°), where the water temperature is warm. Therefore, understanding how selection shapes genetic variations and structure in farmed populations is of paramount importance in evolutionary biology. High-throughput sequencing to generate genome-wide data for fish species allows for elucidating the genomic basis of adaptation to local or farmed conditions and uncovering genes that control the phenotypes of interest. The present study aimed to detect genomic selection signatures and analyze the genetic variability in farmed populations of tambaqui in South America using single-nucleotide polymorphism (SNP) markers obtained with double-digest restriction site-associated DNA sequencing. Initially, 199 samples of tambaqui farmed populations from different locations (located in Brazil, Colombia, and Peru), a wild population (Amazon River, Brazil), and the base population of a breeding program (Aquaculture Center, CAUNESP, Jaboticabal, SP, Brazil) were genotyped. Observed and expected heterozygosity was 0.231-0.350 and 0.288-0.360, respectively. Significant genetic differentiation was observed using global FST analyses of SNP loci (FST = 0.064, p < 0.050). Farmed populations from Colombia and Peru that differentiated from the Brazilian populations formed distinct groups. Several regions, particularly those harboring the genes of significance to aquaculture, were identified to be under positive selection, suggesting local adaptation to stress under different farming conditions and management practices. Studies aimed at improving the knowledge of genomics of tambaqui farmed populations are essential for aquaculture to gain deeper insights into the evolutionary history of these fish and provide resources for the establishment of breeding programs.

Ichthyoplankton DNA metabarcoding: Challenges and perspectives.

DNA metabarcoding has been widely used to access and monitor species. However, several challenges remain open for its mainstream application in ecological studies, particularly when dealing with a quantitative approach. In a From the Cover article in this issue of Molecular Ecology, Mariac et al. (2021) report species-level ichthyoplankton dynamics for 97 fish species from two Amazon river basins using a clever quantitative metabarcoding approach employing a probe capture method. They clearly show that most species spawned during the floods, although ~20% also spawned mainly during the receding period and some other year-round, but interestingly, species from the same genus reproduced in distinct periods (i.e., inverse phenology). Opportunistically, Mariac et al. (2021) reported that during an intense hydrological anomaly, several species had a sharp reduction in spawning activity, demonstrating a quick response to environmental cues. This is an interesting result since the speed at which fish species can react to environmental changes, during the spawning period, is largely unknown. Thus, this study brings remarkable insights into basic life history information that is imperative for proposing strategies that could lead to a realistic framework for sustainable fisheries management practices and conservation, fundamental for an understudied and threatened realm, such as the Amazon River basin.

Integrating Cytogenetics and Population Genomics: Allopatry and Neo-Sex Chromosomes May Have Shaped the Genetic Divergence in the Erythrinus erythrinus Species Complex (Teleostei, Characiformes).

Diversity found in Neotropical freshwater fish is remarkable. It can even hinder a proper delimitation of many species, with the wolf fish Erythrinus erythrinus (Teleostei, Characiformes) being a notable example. This nominal species shows remarkable intra-specific variation, with extensive karyotype diversity found among populations in terms of different diploid chromosome numbers (2n), karyotype compositions and sex chromosome systems. Here, we analyzed three distinct populations (one of them cytogenetically investigated for the first time) that differed in terms of their chromosomal features (termed karyomorphs) and by the presence or absence of heteromorphic sex chromosomes. We combined cytogenetics with genomic approaches to investigate how the evolution of multiple sex chromosomes together with allopatry is linked to genetic diversity and speciation. The results indicated the presence of high genetic differentiation among populations both from cytogenetic and genomic aspects, with long-distance allopatry potentially being the main agent of genetic divergence. One population showed a neo-X1X2Y sexual chromosome system and we hypothesize that this system is associated with enhanced inter-population genetic differentiation which could have potentially accelerated speciation compared to the effect of allopatry alone.

Sublethal effects of environmental concentrations of caffeine on a neotropical freshwater fish.

Caffeine is a contaminant frequently detected in water bodies. Growth trends in both human population and caffeine consumption per capita are expected to exacerbate the occurrence of caffeine in freshwaters. Yet the effects of caffeine on native fish fauna are poorly understood. We exposed larvae of an endemic Neotropical catfish (Rhamdia quelen) to a range of caffeine concentrations for 30 days. We found that larvae exposed to the highest concentration (16 mg L-1) showed skeletal deformations and reduced growth. We further compiled measured environmental concentrations of caffeine in surface freshwater globally and performed a risk assessment. Our analysis points to a low risk to R. quelen and equally sensitive fish species in ~90% of the freshwater ecosystems considered in our analysis. The risk quotient is higher in freshwater ecosystems of South and Central America, where R. quelen is endemic. Although the ecotoxicological risk is currently low in most places, increased caffeine consumption, exacerbated by the lack of sanitation, is expected to increase caffeine concentrations in many parts of the world, posing a threat of sublethal morphological effects to local fish species.

Effects of melatonin supplementation on the quality of cryopreserved sperm in the neotropical fish Prochilodus lineatus.

The cryopreservation process causes damage to sperm structures and supplementation of the cryoprotective medium is an alternative to reduce these damages. The aim of this study was to determine the effect of melatonin supplementation on post-thaw sperm quality in Prochilodus lineatus. The cryoprotective medium was supplemented with 2.00, 2.75, 3.50, and 4.25 mM melatonin, and the control group (without melatonin). Sperm motility parameters, membrane integrity, sperm morphology, oxidative stress (lipid peroxidation and enzymatic activity), and fertilization capacity were analyzed in the post-thaw sperm. Samples cryopreserved with 2.00 mM melatonin yielded higher sperm motility rate than other treatments with the addition of melatonin. Sperm curvilinear velocity (VCL) and average path velocity (VAP) were higher in samples containing 2.00 mM melatonin than in other treatments. Samples from control and with 2.00 mM melatonin presented higher membrane integrity and morphological normality than samples containing 4.25 and 2.75 mM melatonin, respectively. Regarding oxidative stress, lower lipid peroxidation (LPO) occurred in 2.75 and 3.50 mM melatonin, compared to control. While higher enzyme activity of catalase (CAT) occurred in the control than in other treatments, no differences were observed in the activity of superoxide dismutase (SOD). Higher fertilization and hatching rates occurred at 2.75 mM melatonin compared to 4.25 mM. Although no significant differences were observed in LPO between the control and samples supplemented with 2 mM melatonin, it was observed that this dosage of melatonin allowed higher VCL and VSL and reduced values of CAT. However, as there are no differences in motility and fertilization rates between the control and the 2 mM concentration, it is suggested that further studies be carried out with lower concentrations of melatonin in order to determine its effectiveness as an antioxidant in the sperm of this species.

Impact of a large dam on reproduction of a non-migratory teleost species, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae) / Impacto de uma grande barragem na reprodução de uma espécie de teleósteo não migradora, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae)

The release of water from the reservoir hypolimnion, lower concentration of oxygen and the anthropogenic regulation of the river flow, could affect the reproduction of fish, especially migratory species. However, little is known about the effects of these changes in water on non-migratory species. In this sense, the reproduction of Acestrohynchus lacustris was evaluated in two sections of São Francisco River, Minas Gerais, Brazil. Section 1, located immediately downstream from Três Marias Dam (18°09'31.65"S and 45°13'36.00"W) and section 2, located at the confluence of the São Francisco and the Abaeté Rivers (18°02'47.78"S and 45°10'57.95"W). For this, we obtained the physico-chemical parameters of water of each study section. Additionally, biometric data and biological indices of all specimens were measured. Fecundity and follicles diameters were measured in females. Temperature, dissolved oxygen and flow showed lower values in section 1. Fish captured in this section, had lower values of GSI in both sexes, and females presented decreased values of fecundity and follicles diameter. This species showed reproductive activity in the two sections analyzed, however, in section 1 where the temperature and dissolved oxygen presented significant lower values, the reproductive capacity of A. lacustris, was negatively affected.

A liberação de água do hipolímnio do reservatório, baixa concentração de oxigênio e a regulação antropogênica do fluxo do rio podem afetar a reprodução de peixes, principalmente espécies migradoras. No entanto, pouco se sabe sobre os efeitos dessas mudanças nas condições da água em espécies não migradoras. A reprodução de Acestrohynchus lacustris foi avaliada em duas seções do rio São Francisco. Seção 1, localizada imediatamente a jusante da barragem de Três Marias e seção 2, localizada na confluência dos rios São Francisco e Abaeté. Para isso, foram obtidos os parâmetros físico-químicos da água de cada seção do estudo. Além disso, dados biométricos e índices biológicos de todos os peixes capturados foram obtidos. Adicionalmente, nas fêmeas foram medidos os diâmetros dos folículos vitelogênicos e a fecundidade. Temperatura, oxigênio dissolvido e fluxo apresentaram valores mais baixos na seção 1. Os peixes capturados nesta seção apresentaram menores valores de IGS em ambos os sexos, e as fêmeas apresentaram menores valores de fecundidade e diâmetro dos folículos. Essa espécie apresentou atividade reprodutiva nas duas seções analisadas, porém, na seção 1, onde os parâmetros da água apresentam piores condições para o processo reprodutivo de peixes, a capacidade reprodutiva de A. lacustris foi afetada negativamente.