Simulated climate change and atrazine contamination can synergistically impair zebrafish testicular function.

Elements that interfere with reproductive processes can have profound impacts on population and the equilibrium of ecosystems. Global warming represents the major environmental challenge of the 21st century, as it will affect all forms of life in the coming decades. Another coexisting concern is the persistent pollution by pesticides, particularly the herbicide Atrazine (ATZ), which is responsible for a significant number of contamination incidents in surface waters worldwide. While it is hypothesized that climate changes will significantly enhance the toxic effects of pesticides, the actual impact of these phenomena remain largely unexplored. Here, we conducted a climate-controlled room experiment to assess the interactive effects of the projected 2100 climate scenario and environmentally realistic ATZ exposures on the reproductive function of male zebrafish. The gonadosomatic index significantly decreased in fish kept in the extreme scenario. Cellular alterations across spermatogenesis phases led to synergic decreased sperm production and increased germ cell sloughing and death. ATZ exposure alone or combined with climate change effects, disrupted the transcription levels of key genes involved in steroidogenesis, hormone signaling and spermatogenesis regulation. An additive modulation with decreased 11-KT production and increased E2 levels was also evidenced, intensifying the effects of androgen/estrogen imbalance. Moreover, climate change and ATZ independently induced oxidative stress, upregulation of proapoptotic gene and DNA damage in post-meiotic germ cell, but the negative effects of ATZ were greater at extreme scenario. Ultimately, exposure to simulated climate changes severely impaired fertilization capacity, due to a drastic reduction in sperm motility and/or viability. These findings indicate that the future climate conditions have the potential to considerably enhance the toxicity of ATZ at low concentrations, leading to significant deleterious consequences for fish reproductive function and fertility. These may provide relevant information to supporting healthcare and environmental managers in decision-making related to climate changes and herbicide regulation.

Effects of climate change and mixtures of pesticides on the Amazonian fish Colossoma macropomum.

Several studies highlighted the complexity of mixing pesticides present in Amazonian aquatic environments today. There is evidence that indicates that ongoing climate change can alter the pattern of pesticide use, increasing the concentration and frequency of pesticide applications. It is known that the combination of thermal and chemical stress can induce interactive effects in aquatic biota, which accentuates cell and molecular damage. However, considering that the effects of climate change go beyond the increase in temperature the objective of this study was to evaluate the effect of climate change scenarios proposed by 6 th IPCC report and a mixture of pesticides on the tambaqui (Colossoma macropomum). The hypothesis of this study is that the negative effects will be accentuated by the combination of an extreme climate changes scenario and a mixture of pesticides. To test the hypothesis, juvenile tambaqui were exposed to a combination of four pesticides (chlorpyrifos, malathion, carbendazim and atrazine) in two scenarios, one that simulates current environmental conditions and another that predicted the environmental scenario for the year 2100. Fish were subjected to the experimental conditions for 96 h. At the end of the experiment, samples of blood, gills, liver, brain, and muscle were obtained for hematological, genotoxic, biochemical, and histopathological analyses. The results demonstrate that environmentally realistic concentrations of pesticides, when mixed, can alter the biochemical responses of tambaqui. The extreme scenario promotes hematological adjustments, but impairs branchial antioxidant enzymes. There is an interaction between the mixture of pesticides and the extreme scenario, accentuating liver tissue damage, which demonstrates that even increased activity of antioxidant and biotransformation enzymes were not sufficient to prevent liver damage.

In vivo exposure to high temperature compromises quality of the sperm in Colossoma macropomum.

Increases in temperature can affect the reproduction of fish by decreasing the quality of gametes for fertilization. Therefore, this study aimed to evaluate the in vivo effect of temperature on the production and sperm quality of Colossoma macropomum, which is an economically relevant species for Brazil, and other countries. Broodstock were exposed for 10 days at temperatures of 29 °C (n = 4) and 35 °C (n = 4). After exposure, semen was collected and sperm quality was evaluated for kinetic, biochemical, morphological parameters, membrane integrity, and oxygen consumption. The sperm quality of males of specimens of C. macropomum exposed in vivo to the higher temperature was compromised and showed a difference in all the analyses (P < 0.05). Sperm showed decreased motility (51.9 ± 11.6 s) compared to the control (61.3 ± 5.5 s); curvilinear (CLV), straight line (SLV), and average path (APV) velocities decreased, and straightness (STR) and beat cross frequency (BCF) increased; oxygen consumption decreased (32.8 ± 3.4 pmol. (s.ml)-1, picomol per second per ml) compared to the control (46.6 ± 3.5 pmol. (s.ml)-1); and the activity of catalase (CAT) and glutathione S-transferase (GST) enzymes increased and decreased, respectively, in sperm from fish exposed to high temperatures. Furthermore, an increase in non-viable sperm with damaged membranes and sperm with altered morphology was observed. The observed alterations indicate that C. macropomum confined in environments with high temperatures possibly produces more sperm that are unviable for the fertilization process.

Ammonia Increases the Stress of the Amazonian Giant Arapaima gigas in a Climate Change Scenario.

Ammonia is toxic to fish, and when associated with global warming, it can cause losses in aquaculture. In this study, we investigated the physiological and zootechnical responses of Arapaima gigas to the current scenarios and to RCP8.5, a scenario predicted by the IPCC for the year 2100 which is associated with high concentrations of environmental ammonia (HEA). Forty-eight chipped juvenile A. gigas were distributed in two experimental rooms (current scenario and RCP8.5) in aquariums with and without the addition of ammonia (0.0 mM and 2.44 mM) for a period of 30 days. The HEA, the RCP8.5 scenario, and the association of these factors affects the zootechnical performance, the ionic regulation pattern, and the levels of ammonia, glucose, triglycerides, sodium, and potassium in pirarucu plasma. The branchial activity of H+-ATPase was reduced and AChE activity increased, indicating that the species uses available biological resources to prevent ammonia intoxication. Thus, measures such as monitoring water quality in regard to production, densities, and the feed supplied need to be more rigorous and frequent in daily management in order to avoid the accumulation of ammonia in water, which, in itself, proved harmful and more stressful to the animals subjected to a climate change scenario.

Bacterioplankton Communities in Dissolved Organic Carbon-Rich Amazonian Black Water.

The Amazon River basin sustains dramatic hydrochemical gradients defined by three water types: white, clear, and black waters. In black water, important loads of allochthonous humic dissolved organic matter (DOM) result from the bacterioplankton degradation of plant lignin. However, the bacterial taxa involved in this process remain unknown, since Amazonian bacterioplankton has been poorly studied. Its characterization could lead to a better understanding of the carbon cycle in one of the Earth's most productive hydrological systems. Our study characterized the taxonomic structure and functions of Amazonian bacterioplankton to better understand the interplay between this community and humic DOM. We conducted a field sampling campaign comprising 15 sites distributed across the three main Amazonian water types (representing a gradient of humic DOM), and a 16S rRNA metabarcoding analysis based on bacterioplankton DNA and RNA extracts. Bacterioplankton functions were inferred using 16S rRNA data in combination with a tailored functional database from 90 Amazonian basin shotgun metagenomes from the literature. We discovered that the relative abundances of fluorescent DOM fractions (humic-, fulvic-, and protein-like) were major drivers of bacterioplankton structure. We identified 36 genera for which the relative abundance was significantly correlated with humic DOM. The strongest correlations were found in the Polynucleobacter, Methylobacterium, and Acinetobacter genera, three low abundant but omnipresent taxa that possessed several genes involved in the main steps of the ß-aryl ether enzymatic degradation pathway of diaryl humic DOM residues. Overall, this study identified key taxa with DOM degradation genomic potential, the involvement of which in allochthonous Amazonian carbon transformation and sequestration merits further investigation. IMPORTANCE The Amazon basin discharge carries an important load of terrestrially derived dissolved organic matter (DOM) to the ocean. The bacterioplankton from this basin potentially plays important roles in transforming this allochthonous carbon, which has consequences on marine primary productivity and global carbon sequestration. However, the structure and function of Amazonian bacterioplanktonic communities remain poorly studied, and their interactions with DOM are unresolved. In this study, we (i) sampled bacterioplankton in all the main Amazon tributaries, (ii) combined information from the taxonomic structure and functional repertory of Amazonian bacterioplankton communities to understand their dynamics, (iii) identified the main physicochemical parameters shaping bacterioplanktonic communities among a set of >30 measured environmental parameters, and (iv) characterized how bacterioplankton structure varies according to the relative abundance of humic compounds, a by-product from the bacterial degradation process of allochthonous DOM.

Correlation between caffeine and coprostanol in contrasting Amazonian water bodies.

The evaluation of contamination by domestic sewage is relevant in the Amazon region; however, it has neither been well-developed nor accompanied by research or monitoring programs. In this study, caffeine and coprostanol as indicators of sewage were investigated in water samples from Amazonian water bodies that crisscross the city of Manaus (Amazonas state, Brazil) and cover regions with distinct main land uses such as high-density residential, low-density residential, commercial, industrial, and environmental protection areas. Thirty-one water samples were studied based on their dissolved and particulate organic matter (DOM and POM) fractions. Quantitative determination of both caffeine and coprostanol was carried out using LC-MS/MS with APCI in the positive ionization mode. The streams of the urban area of Manaus had the highest concentrations of caffeine (1.47-69.65 µg L-1) and coprostanol (2.88-46.92 µg L-1). Samples from the peri-urban Tarumã-Açu stream and from the streams in the Adolpho Ducke Forest Reserve showed much lower concentrations of caffeine (20.20-165.78 ng L-1) and coprostanol (31.49-120.44 ng L-1). Samples from the Negro River showed a wider range of concentrations of caffeine (20.59-873.59 ng L-1) and coprostanol (31.72-706.46 ng L-1), with the highest values found in the outfalls of the urban streams. Levels of caffeine and coprostanol were significantly positively correlated in the different organic matter fractions. The coprostanol/(coprostanol + cholestanol) ratio proved to be a more suitable parameter than the coprostanol/cholesterol one in low-density residential areas. Proximity to densely populated areas and the flow of water bodies appear to influence the caffeine and coprostanol concentrations, which was observed in their clustering in the multivariate analysis. The results indicate that caffeine and coprostanol can be detected even in water bodies that receive very low domestic sewage input. Therefore, this study revealed that both caffeine in DOM and coprostanol in POM represent viable alternatives for use in studies and monitoring programs even in remote areas of the Amazon, where microbiological analyses are often unfeasible.

Vitellogenin Ab structure of the amazonian Arapaima gigas.

The vitellogenin is composed by polypeptides that are precursors of egg yolk proteins that provides embryo and larvae nutrition. The mRNA encoding for vitellogenin Ab (Vtg-Ab; 4,536 bp long and 1,512 amino acids) were obtained by RNA-Seq library sequencing of pirarucu gonads. The Vtg-Ab sequences had high homology with Vtgs of other three teleosts species of the order Osteoglossiformes. The transcript of ovarian Vtg was identified based on structural criteria, and so we classify the Vtg of pirarucu as Vtg-Ab due to the truncated or shortened phosvitin (N-terminal end) and phosvitinless domain (C-terminal end). The Vtg-Ab of pirarucu present two major deletions with 133 amino acids in the Lipovitellin I domain and 89 amino acids in the truncated or shortened Phosvitin domain, both located in the N-terminal end region. The three-dimensional (3-D) structure Vtg-Ab protein shows the presence of a typical 4α-helices bundle protein that runs in anti-parallel. In general, the characterization of Vtg-Ab may be the useful elucidation of the hormonal regulation of vitellogenesis and improve the production of pirarucu for broodstock management in aquaculture and preparation of Vtg antibody production (species-specific) for sex identification.

Tissue distribution of appetite regulation genes and their expression in the Amazon fish Colossoma macropomum exposed to climate change scenario.

Climate change leads to an increase in water acidification and temperature, two environmental factors that can change fish appetite and metabolism, affecting fish population in both wild and aquaculture facilities. Therefore, our study tested if climate change affects gene expression levels of two appetite-regulating peptides - Neuropeptide Y (NPY) and Cholecystokinin (CCK) - in the brain of tambaqui, Colossoma macropomum. Additionally, we show the distribution of these genes throughout the body. Amino acid sequences of CCK and NPY of tambaqui showed high similarity with other Characiformes, with the closely related order Cypriniformes, and even with the more distantly related order Salmoniformes. High apparent levels of both peptides were expressed in all brain areas, while expression levels varied for peripheral tissues. NPY and CCK mRNA were detected in all peripheral tissues but cephalic kidney for CCK. As for the effects of climate change, we found that fish exposed to extreme climate scenario (800 ppm CO2 and 4.5 °C above current climate scenario) had higher expression levels of NPY and lower expression levels of CCK in the telencephalon. The extreme climate scenario also increased food intake, weight gain, and body length. These results suggest that the telencephalon is probably responsible for sensing the metabolic status of the organism and controlling feeding behavior through NPY, likely an orexigenic hormone, and CCK, which may act as an anorexigenic hormone. To our knowledge, this is the first study showing the effects of climate change on the endocrine regulation of appetite in an endemic and economically important fish from the Amazon. Our results can help us predict the impact of climate change on both wild and farmed fish populations, thus contributing to the elaboration of future policies regarding their conservation and sustainable use.

Impact of high temperature, CO2 and parasitic infection on inflammation, immunodepression and programmed cell death in Colossoma macropomum at the transcriptional level.

The production of tambaqui Colossoma macropomum has recently reached a milestone, being considered the main native species produced in South American continental waters. Despite the importance of this fish, its immunity is poorly understood, and global warming could pose severe risks to its health as increasing water temperature leads to an increase in the incidence of parasitic diseases. In an experimental context based on the high-emission scenario of the 5th Intergovernmental Panel on Climate Change (IPCC) report, we evaluated the synergistic effect of exposure to the extreme climate change scenario (RCP8.5) during two exposure periods (7 and 30 days) and two levels of parasitism by monogeneans (low and high). The goal was to understand how the tambaqui immune system will react to this challenge. To achieve this goal, we analyzed the expression of nine immunity-related genes (jak3, stat3, il-10, socs1, casp1, il-1ß, tp53, bcl2, and hif-1α) in the spleen. Our main findings showed downregulation in the jak3/stat3 pathway, genes related to the control of inflammation and apoptosis, in addition to upregulation of proinflammatory genes and those related to pyroptosis during the first 7 days of exposure to the extreme climate scenario, also indicating a stage of immunodepression in these animals. After 30 days of exposure, all genes tended to return to similar levels in the current scenario, possibly due to the decrease in parasite load caused by chronic exposure to the extreme scenario. Our data strongly suggest that the increase in parasitism intensity caused by the extreme climate change scenario is responsible for disturbances in the host's immune system. However, more studies are needed to clarify this poorly understood cascade of events.

Climate change affects the parasitism rate and impairs the regulation of genes related to oxidative stress and ionoregulation of Colossoma macropomum.

Global climate change represents a critical threat to the environment since it influences organismic interactions, such as the host-parasite systems, mainly in ectotherms including fishes. Rising temperature and CO2 are predicted to affect this interaction other and critical physiological processes in fish. Herein, we investigated the effects of different periods of exposure to climate change scenarios and to two degrees of parasitism by monogeneans in the host-parasite interaction, as well as the antioxidant and ionoregulatory responses of tambaqui (Colossoma macropomum), an important species in South American fishing and aquaculture. We hypothesized that temperature and CO2 changes in combination with parasite infection would interfere with the host's physiological processes that are related to oxidative stress and ionoregulation. We experimentally exposed C. macropomum to low and high levels of parasitism in the current and extreme climate scenarios (4.5 °C and 900 ppm CO2 above current levels) for periods of seven and thirty days and we use as analyzed factors; the exposure time, the climate scenario and parasitism level in a 2 × 2 × 2 factorial through a three-way ANOVA as being fish the experimental unit (n = 8). An analysis of gill enzymatic and gene expression profile was performed to assess physiological (SOD, GPx and Na+/K+-ATPase enzymes) and molecular (Nrf2, SOD1, HIF-1α and NKA α1a genes) responses. A clear difference in the parasitism levels of individuals exposed to the extreme climate scenario was observed with a rapid and aggressive increase that was higher after 7 days of exposure though showed a decrease after 30 days. The combination of exposure to the extreme climate change scenario and parasitism caused oxidative stress and osmoregulatory disturbance, which was observed through the analysis of gene expression (Nrf2, SOD1, HIF-1α and NKA α1a) and antioxidant and ionoregulatory enzymes (SOD, GPx and Na+/K+-ATPase) on the host, possibly linked to inflammatory processes caused by the high degree of parasitism. In the coming years, these conditions may result in losses of performance for this species, and as such will represent ecological damage and economical losses, and result in a possible vulnerability in relation to food security.

Involvement of purinergic system and electron transport chain in two species of cichlids from the Amazon basin exposed to hypoxia.

The Amazonian aquatic ecosystem undergoes seasonal variations and daily changes that directly affect the availability of oxygen. During the day the levels of oxygen can reach supersaturation, and at night can drop to zero. In this way, aquatic organisms are exposed daily to physiological challenges regarding the availability of oxygen. The present study revealed significant differences in the physiology and performance of two cichlids: Geophagus proximus (black water cichlid - from Negro River) and Chaetobranchopsis orbicularis (white water cichlid - from Amazon River), exposed to hypoxia. The white water cichlid showed lower value (1.99 ± 0.79 pKa) of critical pressure of oxygen (Pcrit) and a longer time (68.00 ± 14.11 min) for total loss of balance (LOE); however, this species showed 50% mortality during exposure to hypoxia, while the black water cichlid did not show mortality. Both cichlids presented a decrease in O2 consumption rate (OCR) during hypoxia.. In this sense, it was observed that the black water cichlid presented several physiological strategies during hypoxia, such as, a significant increase in plasma cortisol levels, nucleoside triphosphate diphosphohydrolase activity (for adenosine diphosphate (ADP) as a substrate) in the gills, and the activity of adenosine deaminase (ADA) in gills and liver, in addition to a significant increase in the activity of complexes (II-III) in the transporter chain of electrons in both analyzed tissues and succinate dehydrogenase activity of gills' mitochondria. On the other hand, the only physiological change observed in the white water cichlid was a significant reduction in the activity of complexes II-III in gills and liver. Based on our findings, we can hypothesize that the white water cichlid specie has less tolerant to hypoxia when compared to the black water cichlid.

Copper and cadmium impair sperm performance, fertilization and hatching of oocytes from Amazonian fish Colossoma macropomum.

The contamination of aquatic environments by transition metals can have a direct influence on the reproductive process of several organisms in the aquatic biota. This study aimed to evaluate the effect of cadmium and copper on the sperm of tambaqui (Colossoma macropomum). Male (n = 4) and female (n = 4) specimens of C. macropomum were induced to spermiation and ovulation, with sperm being activated in the following media: 0; 0.6; 1.2 and 1.8 mg/L of cadmium (CdCl2) and 0; 0.4; 0.8 and 1.2 mg/L of copper (CuCl2). Sperm quality was assessed through time (s) and motility rate (%), superoxide dismutase (SOD) and glutathione S-transferase (GST) activities, lipoperoxidation levels (LPO), and morphological characteristics. In parallel, the effects of these metals on the rate of fertilization and hatching of the oocytes were evaluated. The duration and motility rate of sperm were longer in the control treatment, 85.67 ± 11.01 s; 90 ± 0.01%, and progressively decreased to 44.67 ± 4.16 s and 60 ± 5%, respectively, in concentrations of 1.8 mg/L (44.67 ± 4.16 s; 60 ± 5%) of CdCl2 and to 65.67 ± 3.30 s; 70 ± 5%, respectively, in concentrations of 0.8 mg/L of CuCl2. We observed an increase in the activity of the SOD enzyme in sperm cells exposed to 1.2 mg/L of CdCl2. The LPO levels were increased significantly in sperm cells exposed to 1.2 and 1.8 mg/L of CdCl2 and 0.8 mg/L of CuCl2. Fertilization and hatching were severely impaired in the presence of Cd and Cu. These data indicate that environments contaminated with cadmium and copper harm the gametes of C. macropomum.

Aerobic Metabolism Impairment in Tambaqui (Colossoma macropomum) Juveniles Exposed to Urban Wastewater in Manaus, Amazon.

The main purpose of the present study was to investigate the potential use of metabolic parameters as non-specific biomarkers of pollution. The Igarapé do Quarenta is a small urban river crossing an industrial area in the city of Manaus, Amazon, and receives the city wastewater without treatment. The fish tambaqui (Colossoma macropomum) were exposed to water collected from two different sites of that stretch for 96 h. After exposure, routine metabolic rate (RMR) was measured, and fish were euthanized for measurements of electron transport system (ETS) activity, Copper (Cu) and Cadmium (Cd) bioaccumulation and biliary PAHs. Water in the sampling points presented low oxygen and high pH, conductivity, dissolved ions, Cu, Cd and ammonia. Bile concentrations of PAHs were high suggesting industrial pollution. The tambaqui exposed to water from Igarapé do Quarenta showed increased RMR and decreased ETS/RMR suggesting impairment of metabolic fish performance and the potential use of these parameters as biomarkers.

Understanding the Science Surrounding Environmental Consequences and Rehabilitation Actions Stemming from Brazil's Fundão Tailing Dam Rupture.

Collapse of the Fundão tailings dam in November 2015 in Minas Gerais, Brazil, resulted in the release of approximately 36 million cubic meters of primarily clay and fine sand-sized particles to the environment. The spill event extended over 600 km of the Rio Doce catchment and affected terrestrial and aquatic ecosystems and compromised socioeconomic activities throughout the watershed. Numerous monitoring programs and research investigations in the catchment since that time contribute to better understanding of the environmental consequences and plans for rehabilitation of the landscape. The Society of Environmental Toxicology and Chemistry (SETAC) Latin America Geographic Unit hosted the Fundão Dam Rupture Science Meeting in Brasilia, Brazil, in June 2019 to report on the findings from these studies. Fourteen technical papers from that meeting are presented in this Integrated Environmental Assessment and Management (IEAM) special series. These papers and the technical discussions at the meeting reflect the consensus opinions of scientists who attended the symposium. Delegates generally agreed that society must commit to rehabilitation after disasters based on the best available evidence describing the structure and function of affected ecosystems. Scientists can play a crucial role in prioritizing and facilitating rehabilitation actions, as well as monitoring for progress toward achieving goals. Integr Environ Assess Manag 2020;16:569-571. © 2020 SETAC.

O colapso da barragem de rejeitos de Fundão em novembro de 2015 em Minas Gerais, Brasil, resultou na liberação de aproximadamente 36 milhões de metros cúbicos de partículas do tamanho de argila e areia fina para o meio ambiente. O derramamento se estendeu por mais de 600 km na bacia hidrográfica do Rio Doce, afetou os ecossistemas terrestres e aquáticos e comprometeu as atividades socioeconômicas em toda a bacia hidrográfica. Numerosos programas de monitoramento e pesquisas na bacia, desde então, contribuem para uma melhor compreensão das consequências ambientais e para os planos de reabilitação da paisagem. A Sociedade de Toxicologia e Química Ambiental (SETAC) da América Latina sediou o Encontro Científico sobre a Ruptura da Barragem do Fundão em Brasília, Brasil, em junho de 2019, para relatar os resultados desses estudos. Quatorze artigos técnicos dessa reunião são apresentados nesta série especial da revista Integrated Environmental Assessment and Management (IEAM). Esses artigos e as discussões técnicas do encontro refletem as opiniões consensuais dos cientistas que participaram do simpósio. Os delegados em geral concordaram que a sociedade deve se comprometer com a reabilitação após desastres com base na melhor evidência disponível que descreva a estrutura e a função dos ecossistemas afetados. Os cientistas podem desempenhar um papel crucial na priorização e facilitação de ações de reabilitação, bem como no acompanhamento da evolução do cumprimento das metas. Integr Environ Assess Manag 2020;16:569-571.

Fish Skin and Gut Microbiomes Show Contrasting Signatures of Host Species and Habitat.

Teleost fish represent an invaluable repertoire of host species to study the factors shaping animal-associated microbiomes. Several studies have shown that the phylogenetic structure of the fish gut microbiome is driven by species-specific (e.g., host ancestry, genotype, or diet) and habitat-specific (e.g., hydrochemical parameters and bacterioplankton composition) factors. However, our understanding of other host-associated microbial niches, such as the skin mucus microbiome, remains limited. The goal of our study was to explore simultaneously the phylogenetic structure of the fish skin mucus and gut microbiome and compare the effect of species- and habitat-specific drivers on the structure of microbial communities in both tissues. We sampled 114 wild fish from 6 populations of 3 ecologically and phylogenetically contrasting Amazonian teleost species. Water samples were collected at each site, and 10 physicochemical parameters were characterized. The skin mucus, gut, and water microbial communities were characterized using a metabarcoding approach targeting the V3-V4 regions of the 16S rRNA. Our results showed a significant distinction between the phylogenetic profile and diversity of the microbiome from each microbial niche. Skin mucus and bacterioplankton communities were significantly closer in composition than gut and free-living communities. Species-specific factors mostly modulated gut bacterial communities, while the skin mucus microbiome was predominantly associated with environmental physicochemistry and bacterioplankton community structure. These results suggest that the variable skin mucus community is a relevant target for the development of microbial biomarkers of environmental status, while the more conserved gut microbiome is better suited to study long-term host-microbe interactions over evolutionary time scales.IMPORTANCE Whether host-associated microbiomes are mostly shaped by species-specific or environmental factors is still unresolved. In particular, it is unknown to what extent microbial communities from two different host tissues from the same host respond to these factors. Our study is one of the first to focus on the microbiome of teleost fish to shed a light on this topic as we investigate how the phylogenetic structure of microbial communities from two distinct fish tissues are shaped by species- and habitat-specific factors. Our study showed that in contrast to the teleost gut microbiome, skin mucus communities are highly environment dependent. This result has various implications: (i) the skin mucus microbiome should be used, rather than the gut, to investigate bacterial biomarkers of ecosystem perturbance in the wild, and (ii) the gut microbiome is better suited for studies of the drivers of phylosymbiosis, or the coevolution of fish and their symbionts.

Disturbance of oxidant/antioxidant status and impairment on fillet fatty acid profiles in Brycon amazonicus subjected to acute heat stress.

Amazon fish are vulnerable to climate change. Current temperature increases in Amazonian rivers are likely to continue in the coming years. Elevated temperatures impair homeostasis and subject fish to oxidative stress; nevertheless, the effects of elevated temperature on plasma and muscle oxidative stress as well as fillet fatty acid composition in matrinxã (Brycon amazonicus) remain unknown. Therefore, the aims of the present study were to determine whether antioxidant system is able to withstand acute thermal stress to avoid plasma and muscle oxidative damage and to determine the manner in which matrinxã adjust their muscle fatty acid metabolism in a response to acute heat stress. We exposed juvenile matrinxã to four temperatures (28 °C as control and 30, 32, and 34 °C) for 72 h and observed the effects on plasma and muscle oxidant/antioxidant status and on fatty acid composition. Plasma reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in matrinxã exposed to 34 °C compared with the control group, while they were significantly higher in the muscle of matrinxã exposed to 32 °C and 34 °C compared with the control group. Superoxide dismutase (SOD) activity and total antioxidant capacity (TAC) levels were significantly lower in the plasma and muscle of matrinxã exposed to 34 °C compared with the control group. Plasma glutathione peroxidase (GPx) activity was significantly lower in matrinxã exposed to 32 and 34 °C compared with the control group, while it was significantly lower in the muscle of matrinxã exposed to 34 °C. The total content of saturated fatty acid (SFA) was significantly higher in the fillet of matrinxã exposed to 34 °C compared with the control group, while the total content of polyunsaturated fatty acid (PUFA) was significantly lower. Based on this evidence, we conclude that acute heat stress at 32 °C and 34 °C causes plasma and muscular oxidative stress, and the enzymatic and non-enzymatic antioxidant system is unable to prevent oxidative damage. Moreover, increased levels of total SFA and decreased levels of PUFA occur in an attempt to withstand heat stress and maintain membrane fluidity; nevertheless, these responses can represent negative impacts for consumers.

Purine levels and purinergic signaling in plasma and spleen of Brycon amazonicus exposed to acute heat thermal stress: An attempt to regulate the immune response.

Amazon fish are vulnerable to climate change. Several lines of evidence suggest that the temperature of Amazonian rivers will increase in the coming years. Elevated temperature disturbs homeostasis and subjects fish to physiological stress; however, the effects of temperature on immunity remain poorly understood, particularly those effects involving purinergic signaling. This system fine-tunes the inflammatory and immune responses triggered by stress. Therefore, the aims of this study were to determine whether acute heat stress induces the release of nucleotides into extracellular compartment and to determine whether purinergic enzymes modulate the proinflammatory effects of adenosine triphosphate (ATP) in plasma and spleen of matrinxã (Brycon amazonicus) exposed to acute heat stress. We exposed juvenile matrinxã to four temperature regimes (28 °C as control, 30, 32 and 34 °C) for 72 h and observed the effects on purinergic signaling. Plasma cortisol levels were significantly higher in fish exposed to 34 °C than in the control group, while spleen ATP, adenosine diphosphate (ADP) and adenosine monophosphate (AMP) levels were significantly higher in this group than in controls. Activities of spleen nucleoside triphosphate diphosphohydrolase (NTPDase) and 5'-nucleotidase were significantly higher in fish exposed to 34 °C than those of the control group, while spleen interleukin-1 (IL-1) and interleukin-6 (IL-6) levels were higher in this same group than in the control group. No significant differences were observed between the groups regarding plasma parameters. Based on these data, we concluded that acute heat stress at 34 °C caused physiological stress in matrinxã, manifesting as elevated plasma cortisol levels. The most important finding is that purinergic enzymes were modulated, though not efficiently, in response to the excessive release of nucleotides into the extracellular space. In summary, the purinergic signaling pathway may be involved in the impairment of immune and inflammatory responses in matrinxã exposed acutely to 34 °C.

How will farmed populations of freshwater fish deal with the extreme climate scenario in 2100? Transcriptional responses of Colossoma macropomum from two Brazilian climate regions.

Tambaqui (Colossoma macropomum Cuvier, 1818) is an endemic fish of the Amazon and Orinoco basins, and it is the most economically important native species in Brazil being raised in five climatically distinct regions. In the face of current global warming, environmental variations in farm ponds represent additional challenges that may drive new adaptive regional genetic variations among broodstocks of tambaqui. In an experimental context based on the high-emission scenario of the 5th Intergovernmental Panel on Climate Change (IPCC) report, we used two farmed tambaqui populations to test this hypothesis. RNA-seq transcriptome analysis was performed in the liver of juvenile tambaqui from northern (Balbina Experimental Station, Balbina, AM) and southeastern (Brumado Fish Farming, Mogi Mirim, SP) Brazilian regions kept for 30 days in artificial environmental rooms mimicking the current and extreme climate scenarios. Three Illumina MiSeq runs produced close to 120 million 500 bp paired-end reads; 191,139 contigs were assembled with N50 = 1595. 355 genes were differentially expressed for both populations in response to the extreme scenario. After enrichment analysis, each population presented a core set of genes to cope with climate change. Northern fish induced genes related to the cellular response to stress, activation of MAPK activity, response to unfolded protein, protein metabolism and cellular response to DNA damage stimuli. Genes biologically involved in regulating cell proliferation, protein stabilisation and protein ubiquitination for degradation through the ubiquitin-proteasome system were downregulated. Genes associated with biological processes, including the cellular response to stress, MAPK cascade activation, homeostatic processes and positive regulation of immune responses were upregulated in southeastern fish. The downregulated genes were related to cytoskeleton organisation, energy metabolism, and the regulation of transcription and biological rhythms. Our findings reveal the signatures of promising candidate genes involved in the regional plasticity of each population of tambaqui in dealing with upcoming climate changes.

Extreme climate scenario and parasitism affect the Amazonian fish Colossoma macropomum.

Global warming caused by greenhouse gases accumulation, in particular carbon dioxide, is the major current environmental challenge, as it will affect all life forms over the next decades. Aquaculture, a fast growing food production sector, is already facing the effects of global warming. The fish immune system is expected to be especially affected by increased temperature and carbon dioxide, mainly when associated to infectious diseases outbreaks. Here, we experimentally investigated the associated effects of an extreme climate scenario and two levels of monogenean parasitism on the hematological and immunological response of an important Amazon fish for continental aquaculture: Colossoma macropomum. Individuals of C. macropomum with low and high levels of parasitism were exposed to current and extreme climate scenarios (4.5 °C and 900 ppm CO2 above current levels). We characterized their hematological profile using classical methods, and their immune-related gills gene expression (HSP70, IL-1ß and IL-10) using quantitative real-time polymerase chain reaction (qPCR). After 7 days of exposure, we observed that exposure to extreme climate scenario caused rapid increase of parasitism intensity and likely acute inflammation, indicated by the higher expression of HSP70 and IL-1ß. The IL-10 gene was downregulated in both groups exposed to extreme climate scenario, contrasting with animals exposed to current scenario. Thus, in the current scenario, the parasitized tambaqui showed a response to the tissue damage, which was not observed in the animals exposed to the extreme scenario.

Temperature and food availability alters the physiology and aerobic capacity of tambaqui (Colossoma macropomum).

Fish of the Amazon experience both daily and seasonal variation in temperature and food availability. In the present work, we investigated the influence of nutrient status changes resulting from feeding Colossoma macropomum five flooded forest fruits on aerobic and swimming performance. To assess the effects of diet, three groups of fish were provided different types of food. One group of was provided with commercial feed, another was given fruit feed, and the last group was not fed. The effects of temperature were tested by keeping half of each group of animals at either 26 °C or 33°C for 30 days. After this period, the animals were transferred to either a respiratory chamber or swimming tunnel. Then, tissues were collected for relevant analyses. Results showed that C. macropomum depresses aerobic metabolism when food is withheld (27% reduction in MO2), and observed effects were intensified at the highest temperature (40% reduction in MO2 at 33 °C). Further, increased temperature negatively influenced hematological (Ht, Hb, VCM), biochemical (glucose, triglycerides, cholesterol), hepatossomatic index and enzymatic (MDH and CS in white muscle) parameters. The animals feeding commercial and fruits enriched diets, have a physiological and biological advantage when compared to the unfed animals at both temperatures. Metabolic suppression is an important strategy for maintaining swimming capacity in C. macropomum. It is suggested that even after a long period of fasting, carbohydrates are important energetic substrates.