Oil spill in an amazon blackwater environment: Biochemical and physiological responses of local fish species.

The accidental spill of petroleum asphalt cement (PAC) in São Raimundo (SR Harbor, located on the Rio Negro (Manaus, Amazonas, Brazil) was monitored through the analysis of polyciclic aromatic hydrocarbons (PAHs) in water and a set of biomarkers in fishes (exposure biomarkes: PAHs-type metabolites concentrations in bile; the activities of ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in liver. Effect biomarkers: lipid peroxidation concentration (LPO) in liver, acetylcholinesterase activity in brain, and genotoxic DNA damage in erythrocytes). Two fish species, Acarichthys heckelii and Satanoperca jurupari, were collected 10, 45, and 90 days after the PAC spill in São Raimundo. At the same time, fish were collected from the Tupé Sustainable Development Reserve (Tupé) which served as a reference area. The sampling periods were related to the rising waters of the natural flood pulse of the Rio Negro. Higher concentrations of PAHs in water were observed at 10 and 45 days and returned to the values of TP 90 days after the PAC spill, a period in which harbor waters rose about 0.2 m. Unlike the PAHs in water, biomarker responses in both fish species significantly increased following the PAC spill in SR. Hepatic ethoxyresorufin-O-deethylase (EROD), PAH-like metabolites in bile, and erythrocyte DNA damage increases, together with inhibition of acetylcholinesterase (AChE) activity in the brain were the most evident responses for both fish species. The calculated pyrolytic index showed mixed sources of PAHs (petrogenic and pyrolytic). The applied PCA-FA indicated important relationships between dissolved organic carbon (DOC) and PAHs concentrations in water, where DOC and PAHs concentrations contributed to biomarkers responses for both fish species in all collection periods.

Effective practices for thermal tolerance polygon experiments using mottled catfish Corydoras paleatus.

Temperature is an important environmental factor that affects how organisms allocate metabolic resources to physiological processes. Laboratory experiments that determine absolute thermal limits for representative species are important for understanding how fishes are affected by climate change. Critical Thermal Methodology (CTM) and Chronic Lethal Methodology (CLM) experiments were utilized to construct a complete thermal tolerance polygon for the South American fish species, Mottled catfish (Corydoras paleatus). Mottled catfish showed Chronic Lethal Maxima (CLMax) of 34.9 ± 0.52 °C and Chronic Lethal Minima (CLMin) of 3.8 ± 0.08 °C. Fish were chronically acclimated (∼2 weeks) to 6 temperatures ranging from 7.2 ± 0.05 °C →32.2 ± 0.16 °C (7 °C, 12 °C, 17 °C, 22 °C, 27 °C, and 32 °C), and CTM used to estimate upper and lower acute temperature tolerance. Linear regressions of Critical Thermal Maxima (CTMax) and Minima (CTMin) data with each acclimation temperature were used along with CLMax and CLMin to create a complete thermal tolerance polygon. The highest CTMax was 38.4 ± 0.60 °C for fish acclimated to 32.2 ± 0.16 °C, and the lowest CTMin was 3.36 ± 1.84 °C for fish acclimated to 7.2 ± 0.05 °C. Mottled catfish have a polygon measuring 785.7°C2, and the slope of the linear regressions showed the species gained 0.55 °C and 0.32 °C of upper and lower tolerance per degree of acclimation temperature, respectively. We compared slopes of CTMax or CTMin regression lines to each other using a set of comparisons between 3, 4, 5, or 6 acclimation temperatures. Our data demonstrated that 3 acclimation temperatures were as sufficient as 4 â†’ 6 to pair with estimates of chronic upper and lower thermal limits for accurately determining a complete thermal tolerance polygon. Construction of this species' complete thermal tolerance polygon provides a template for other researchers. The following is sufficient to generate a complete thermal tolerance polygon: Three chronic acclimation temperatures that are spread somewhat evenly across a species' thermal range, include an estimation of CLMax and CLMin, and are followed by CTMax and CTMin measurements.

Severe damages caused by Malathion exposure in Colossoma macropomum.

The increase in pesticide use in response to agricultural demands poses a risk to non-target organisms, including fish. Integrated analysis of biochemical, histopathological and genetic parameters in fish exposed to Malathion insecticide provide information on the toxicity mechanisms of this pesticide, which is classified as a probable carcinogen for humans. The present study assessed the biological responses of Colossoma macropomum after exposure to Malathion. We started determining the lethal concentration, which is the concentration capable of killing 50% of the subjects in an acute toxicity test (LC50-96 h), which was 15.77 ± 3.30 mgL-1. The fish were, then, exposed to Malathion during 96 h at a sublethal concentration, 7.30 mgL-1. Overall, we observed an increased activity of biotransformation and antioxidant enzymes, which reduced production of mitochondrial reactive oxygen species after 96 h exposure, as well as kept constant the mitochondrial respiration, Acetylcholinesterase activity and DNA damage. However, fish exposed to insecticide presented severe gill histopathological damage and increased expression of proto-oncogene ras. Taken together, the results suggest that, after four days of exposure to the Malathion, C. macropomum efficiently activates its defense mechanisms, suggesting that the basal response mechanisms are responsive. On the other hand, histopathologic damages evidenced the adverse effects of Malathion on fish, since it promoted gill necrosis and increased the expression of ras oncogene that is directly related to tumorigenesis events.

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.

Experimentally increased temperature and hypoxia affect stability of social hierarchy and metabolism of the Amazonian cichlid Apistogramma agassizii.

The primary goal of this study was to understand how changes in temperature and oxygen could influence social behaviour and aerobic metabolism of the Amazonian dwarf cichlid Apistogramma agassizii. Social hierarchies were established over a period of 96h by observing the social interactions, feeding behaviour and shelter use in groups of four males. In the experimental environment, temperature was increased to 29°C in the high-temperature treatment, and oxygen lowered to 1.0mg·L(-1)O2 in the hypoxia treatment. Fish were maintained at this condition for 96h. The control was maintained at 26°C and 6.6mg·L(-1)O2. After the experimental exposure, metabolism was measured as routine metabolic rate (RMR) and electron transport system (ETS) activity. There was a reduction in hierarchy stability at high-temperature. Aggression changed after environmental changes. Dominant and subdominant fish at high temperatures increased their biting, compared with control-dominant. In contrast, hypoxia-dominant fish decreased their aggressive acts compared with all other fish. Shelter use decreased in control and hypoxic dominant fish. Dominant fish from undisturbed environments eat more than their subordinates. There was a decrease of RMR in fish exposed to the hypoxic environment when compared with control or high-temperature fish, independent of social position. Control-dominant fish had higher RMR than their subordinates. ETS activity increased in fish exposed to high temperatures; however, there was no effect on social rank. Our study reinforces the importance of environmental changes for the maintenance of hierarchies and their characteristics and highlights that most of the changes occur in the dominant position.

Temporal exposure to malathion: Biochemical changes in the Amazonian fish tambaqui, Colossoma macropomum.

The main toxicity mechanism of organophosphate insecticides such as malathion is the acetylcholinesterase enzyme inhibition. However, fish responses to organophosphates may vary depending on the activation of different defense mechanisms as well as the length of exposure. As such, the evaluation of acetylcholinesterase activity, in combination with the evaluation of biotransformation and antioxidants enzymes levels, is useful for indicating damage in fish exposed to this insecticide. Moreover, evaluating mitochondrial activity might evidence how the hierarchic responses occur in relation to the length of time that the fish is exposed. Therefore, the aim of our study is to evaluate whether the length of exposure to malathion differentially affects the biochemical responses of tambaqui. Our hypothesis is that the physiological alterations due to exposure are time dependent. Fish were exposed to sublethal concentrations of the insecticide during 6, 12, 24, 36, and 48 h. Contrary to expectations, there was no acetylcholinesterase activity inhibition during the experiment, which indicates an absence of neurotoxicity. Phase II biotransformation mechanism was activated early, especially in the liver. Oxidative damage was evident in the first hours of exposure and was concurrent with the activation of antioxidant enzymes. Mitochondrial bioenergetics were differentially affected by the length of exposure. The data suggest that the tambaqui regulates mitochondrial respiration differently over time, seeking to maintain homeostasis and ATP demand, and ensures the activation of response mechanisms, thus minimizing oxidative damage and avoiding the neurotoxicity of malathion.

Oxygen-dependent distinct expression of hif-1α gene in aerobic and anaerobic tissues of the Amazon Oscar, Astronotus crassipinnis.

The aquatic habitats of the Amazon basin present dramatic variation of oxygen level, and, to survive such changes, many aquatic animals developed biochemical and physiological adaptations. The advanced teleost Astronotus crassipinnis (Perciformes) is a fish tolerant to hypoxia and known to endure such naturally variable environments. Hypoxia-Inducible factor-1α (hif-1α) is among the most important and studied genes related to hypoxia-tolerance, maintaining regular cellular function and controlling anaerobic metabolism. In the present work, we studied hif-1α expression and related it to changes in metabolic pathways of Astronotus crassipinnis exposed to 1, 3 and 5 h of hypoxia, followed by 3 h of recovery. The results show that A. crassipinnis depresses aerobic metabolic under hypoxia, with a decrease in glycolysis and oxidative enzyme activities, and increases its anaerobic metabolism with an increase in LDH activity coupled with a decrease in oxygen consumption, which indicates an increase in anaerobic capacity. In addition, the animal differentially regulates hif-1α gene in each tissue studied, with a positive relationship to its metabolic profile, suggesting that hif-1α might be one of the most important induction factors that regulate hypoxia tolerance in this species.

Gene expression, genotoxicity, and physiological responses in an Amazonian fish, Colossoma macropomum (CUVIER 1818), exposed to Roundup® and subsequent acute hypoxia.

Roundup® (RD) is a glyphosate-based herbicide used to control weeds in agriculture, and fishponds. In the Amazon, hypoxia is a natural phenomenon in flooded areas. Beyond the challenge of hypoxia, fish need to cope with the use of pesticides as RD that increases in the aquatic environment through the leaching of agricultural areas, and in aquaculture fish tanks. Thus, there is a need to better understand the combined effects of hypoxia and RD contamination for aquatic biota. The aim of this study was to investigate the effects of Roundup® (RD) and subsequent acute hypoxia in the gene expression, genotoxicity, histological and physiological responses of Colossoma macropomum. Fish were individually exposed to four different treatments during 96 h: normoxia (N), hypoxia (H), RD plus normoxia (NRD), and RD plus hypoxia (HRD) (RD concentration represents 75% of LC50 - nominal concentration 15 mg L-1 to C. macropomum). HRD fishes presented down-regulation of hif-1α gene and ras oncogene, while NRD fish presented overexpression of ras; no difference occurred in hif-1α gene expression in both normoxia treatments. The glutathione-S-transferase and catalase activities increased in HRD fish liver compared to NRD. Otherwise, there was no difference in lipoperoxidation (LPO) between all treatments. Genetic Damage Index, measured throughout comet assay in erythrocytes of all treatments, presented similar values, excepted by fish exposed to NRD. As regard as hypoxic exposure, hypoxic fish presented significantly lower values, compared to HRD fishes. An increase in liver histological injuries occurred in H and HRD fish groups. In conclusion, we may affirm that C. macropomum is sensitive concerning RD contamination and that this sensitivity increases when combined with hypoxia.