Influence of acute heat shock on antioxidant defense of tropical fish, Psalidodon bifasciatus.

Psalidodon bifasciatus is a fish species sensitive to physical and chemical changes in water. It serves as a good bioindicator of temperature variations and is utilized in environmental monitoring studies in Brazilian rivers. The objective of this study was to evaluate antioxidant defense biomarkers in the heart, brain, and muscle of P. bifasciatus exposed to a 10 °C thermal increase. P. bifasciatus were collected and divided into a control group (21 °C) and groups subjected to thermal shock (31 °C) for periods of 2, 6, 12, 24, and 48h. Two-way ANOVA indicated that a 10 °C temperature increase caused oxidative stress in P. bifasciatus. This was evidenced by altered levels of lipid peroxidation (LPO), carbonylated proteins (PCO), and glutathione peroxidase (GPx) in the heart, catalase (CAT) and LPO in the brain, and LPO in the muscle. Principal component analysis (PCA) and integrated biomarker response (IBR) analysis indicated that, compared to the heart and muscle, the brain exhibited a greater activation of the antioxidant response. Sensitivity analysis indicated that the muscle was the most sensitive organ, followed by the brain and heart. Our results indicate that the stress response is tissue-specific through the activation of distinct mechanisms. These responses may be associated with the tissue's function as well as its energy demand. As expected, P. bifasciatus showed changes in response to thermal stress, with the brain showing the greatest alteration in antioxidant defenses and the muscle being the most sensitive tissue.

Physiological strategies of acute thermal conditions of Rhamdia voulezi collected in the Iguaçu river watershed, Paraná, Brazil: biochemical markers of metabolic and oxidative stress.

Thermal pollution creates substantial challenges that alter energy demand and produce reactive oxygen species that damage fish DNA, proteins, and lipids. Rhamdia voulezi is a species of fish native to the Iguaçu river, Paraná, Brazil, that does not have scientific records of minimum (CTmin) and maximum (CTmax) temperatures required for survival. As it is a top predator species in the food chain and lives at temperatures below 22 °C, the loss of the species can cause functional problems in controlling the ecosystem and energy flow. The study evaluated the tissue metabolism of the brain, heart, and muscle of R. voulezi (n = 72) subjected to acute thermal stress of 31 °C for 2, 6, 12, 24, and 96 h after acclimatization to 21 °C. The biochemical markers SOD, GPx, MDH, HK, and CK of the brain, PCO of the heart and CAT, glycogen, G6PDH, and ALT of muscle were significant. PCA, IBR, thermal sensitive, and condition factor suggested that R. voulezi has different physiological strategies for acclimatization to 31 °C to mobilize and sustain the metabolic needs of oxygenation and energy allocation/utilization for tissue ATP production.

Metabolic responses in the gills of Yellowtail Lambari Astyanax lacustris under low- and high-temperature thermal stress.

OBJECTIVE: Ectothermic fish are directly affected by temperature changes in the environment. The aim of this study was to evaluate the metabolic responses in the gills of Yellowtail Lambari Astyanax lacustris under thermal stress. METHODS: To this end, we used spectrophotometry to evaluate the biomarkers of carbohydrate and protein metabolism, antioxidant defense, and oxidative damage in fish subjected to low (15°C) and high (31°C) temperatures, with control groups held at 23°C, for 2, 6, 12, 24, 48, and 96 h. RESULT: The results showed that cold thermal stress did not change the energy demand, and the antioxidant defense was reduced; therefore, the gills were vulnerable to the action of reactive oxygen species (ROS), presenting increased protein carbonylation at 12 h. With heat thermal stress, a higher energy demand was observed, which was verified by an increase in aerobic metabolism by glycolysis and the citric acid cycle. High-temperature stress also increased the antioxidant defenses, as verified by the increased activities of glutathione peroxidase, glutathione reductase, and glutathione S-transferase. However, the antioxidant defense system could not protect tissues from the action of ROS, as protein carbonylation increased at 6 and 24 h, indicating oxidative stress. CONCLUSION: The results showed that (1) temperature variations caused metabolic adjustments in the gills of Yellowtail Lambari, (2) the adaptive responses were different for winter and summer temperatures, and (3) Yellowtail Lambari recovered homeostasis when subjected to thermal stress, even with the occurrence of oxidative stress.

Assessing physiological responses and oxidative stress effects in Rhamdia voulezi exposed to high temperatures.

Exposure to high temperatures induces changes in fish respiration, resulting in an increased production of reactive oxygen species. This, in turn, affects the enzymatic and non-enzymatic components of antioxidant defenses, which are essential for mitigating cellular stress. Rhamdia voulezi, an economically important fish species endemic to Brazil's Iguaçu River, served as the subject of our study. Our goal was to assess enzymatic antioxidant biomarkers (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glucose-6-phosphate dehydrogenase), non-protein thiol levels (reduced glutathione), and markers of oxidative damage (lipoperoxidation and carbonylation) in the liver, gills, and kidneys of R. voulezi after acute exposure to high temperatures (31°C) for 2, 6, 12, 24, and 96 h. Control groups were maintained at 21°C. Our findings revealed that the liver exhibited increased superoxide dismutase levels up to 12 h and elevated glutathione S-transferase levels at 12 and 96 h at 31°C. In the gills, superoxide dismutase levels increased up to 24 h, along with increased lipoperoxidation at 2, 6, 12, and 96 h of exposure to high temperatures. The kidneys responded to heat stress at 12 h, with an increase in superoxide dismutase and catalase activity, and lipid peroxidation was observed at 2 and 6 h at 31°C. The three tissues evaluated responded differently to heat stress, with the liver demonstrating greater physiological adjustment to high temperatures. The intricate interplay of various antioxidant defense biomarkers and oxidative damage suggests the presence of oxidative stress in R. voulezi when exposed to high temperatures.

Integrated biomarker response index as an ally in the observation of metabolic biomarkers in muscle of Astyanax lacustris exposed to thermal variation.

Temperature is an important factor that conditions the physiological responses of fish, generating a stressful condition when in non-ideal parameters. Thus, the objective was to evaluate metabolic aspects in the muscle of fish Astyanax lacustris submitted to thermal shock. The specimens were subjected to 2, 6, 12, 24, 48, and 96 h of exposure to 15 °C and 31 1 °C, with their respective controls 23 °C. At 15 °C there was a reduction in glycogenolysis in the initial periods indicated by changes in glycogen phosphorylase (GP), pyruvate and lactate. Subsequently, there was an increase in GP activity, pyruvate levels and hexokinase activity in the next time periods, suggesting an increase in energy demand. At 31 °C there was observed low need for the protein metabolism, indicated by reduction in the activity of aspartate aminotransferase and alanine aminotransferase. At 15 °C, initial periods show an increase in glutathione reductase activity and an increase in carbonylated protein levels, indicating induction of oxidative stress for muscle. At 31 °C, there was a punctual increase in reduced glutathione levels at 24 h. In addition, the integrated biomarker response index proved to be a good ally in the evaluation of a set of biomarkers, corroborating the results observed by the biomarkers individually. Thus, it is possible to conclude that the acute thermal shock affects the metabolism of A. lacustris muscle, which undergoes rearrangements to deal with temperature, where 15 °C is more stressful than 31 °C.

Gradual increase of temperature trigger metabolic and oxidative responses in plasma and body tissues in the Antarctic fish Notothenia rossii.

Antarctica is considered a thermally stable ecosystem; however, climate studies point to increases in water temperatures in this region. These thermal changes may affect the biological processes and promote metabolic changes in the adapted organisms that live in this region, rendering the animals more vulnerable to oxidative damage. This study assessed the effect of acclimation temperature on the levels of stress response markers in plasma, kidney, gill, liver, and brain tissues of Notothenia rossii subjected to gradual temperature changes of 0.5 °C/day until reaching temperatures of 2, 4, 6, and 8 °C. Under the effect of the 0.5 °C/day acclimation rate, gill tissue showed increased glutathione-S-transferase (GST) activity; kidney tissue showed increased H+-ATPase activity. In the liver, there was also an increase in GSH. In plasma, gradual decreases in the concentrations of total proteins and globulins were observed. These responses indicate a higher production of reactive oxygen species ROS, an imbalance in energy demand, and a lack in protein synthesis. Gradual increase in temperature may cause opposite responses to the thermal shock model in N. rossii.

Cold and warm waters: energy metabolism and antioxidant defenses of the freshwater fish Astyanax lacustris (Characiformes: Characidae) under thermal stress.

Subtropical fish are exposed to seasonal variations in temperature that impose a set of adaptations on their metabolism necessary for the maintenance of homeostasis. In this study, we addressed the effects of temperature variation on the metabolism of Astyanax lacustris, a species of freshwater fish common in the subtropical region of Brazil. Biomarkers of carbohydrate and protein metabolism, antioxidant defense, and oxidative damage were evaluated in the liver of A. lacustris exposed to low (15 °C) and high (31 °C) temperature thermal shock, with controls at 23 °C for 2, 6, 12, 24, 48, 72, and 96 h. A high energy demand was observed during the first 48 h of exposure to 15 °C, which is necessary for metabolic adjustment at low temperatures, with an increase in glycolysis, citric acid cycle, and amino acid catabolism. In addition, at 31 °C, glucose was exported in the first 12 h of exposure, and an increase in the citric acid cycle suggested acetyl-CoA as the pathway substrate, originating from the oxidation of lipids. The antioxidant defenses did not change at 15 °C, as opposed to 31 °C, in which there were changes in several antioxidant defense markers, indicating a response to the production of ROS. However, oxidative stress was observed at both temperatures, with oxidative damage detected by lipid peroxidation at 15 °C and protein carbonylation at 31 °C.

Biomarkers of oxidative stress and cell damage in freshwater bivalves Diplodon parodizi exposed to landfill leachate.

Landfill is a public and environmental health problem; establishing and understanding methodologies to decrease its toxicity are thus necessary. Leachate samples were collected, at a sanitary landfill, immediately after the exit from the landfill, i.e. raw leachate (collection point A), after conventional treatment (point B) and after treatment by wetlands (point C). D. parodizi specimens were exposed to 3%, 10% and control (0%) dilutions of leachate from these collection points for 7 days. Markers of antioxidant defences and cell damage were analysed. At point B, the gills of D. parodizi showed higher glutathione-S-transferase (GST) and glutathione reductase (GR) activity; the latter is a supplier of glutathione reductase (GSH). The low GST activity at point A was associated with the hormesis effect. Higher levels of superoxide dismutase (SOD), ethoxyresorufin-O-deethylase (EROD) and glutathione peroxidase (GPx) occurred at point A. Glucose-6-phosphate dehydrogenase (G6PDH) was inhibited at the points with the highest pollutant load and at the highest leachate dilutions. Higher levels of markers at point A may be related to the high pollutant charge and specific compounds present in the untreated leachate. The multi-xenobiotic resistance mechanism (MXR), metallothionein-like proteins (MT) and lipid peroxidation (LPO) did not vary among treatments. The biomarker responses showed negative effects of the leachate on the freshwater bivalve and simultaneously showed that the wetland treatment employed at the Caximba sanitary landfill is effective.

Effect of gradual temperature increase on the carbohydrate energy metabolism responses of the Antarctic fish Notothenia rossii.

The warming of the Southern Ocean waters may affect the biological processes and the performance of the fish inhabiting it. The notothenioid group is metabolically specialized to low-temperature environments and may be vulnerable to the climatic changes imposed on the Antarctic continent. However, gradual temperature changes potentially allow an opportunity for plasticity adjustments. The present study evaluated the effect of gradual increase of temperature on the enzymatic and nonenzymatic parameters of energy metabolism in renal, branchial, hepatic, and encephalic tissue of Notothenia rossii subjected to a gradual temperature change of 0.5 °C/day until reaching 2 °C, 4 °C, 6 °C, and 8 °C. Under the effect of an acclimation rate of 0.5 °C/day, the gill tissue showed increased phosphofructokinase (PFK) enzyme activity. In the kidney, there was increased activity of the malate dehydrogenase (MDH), glucose-6-phosphatase (G6PDH), and glycogen phosphorylase (GP) enzymes. There was an increase in lactate concentration in the liver and an increase in GP enzyme activity in the brain. The specific tissue responses indicate the presence of thermal plasticity and an attempt to regulate energy metabolism to mitigate thermal stress in this species under these experimental conditions, possibly through the activation of glycolysis, gluconeogenesis, and glycogenolysis.

Effect of long-term thermal challenge on the Antarctic notothenioid Notothenia rossii.

The thermal stability of the Antarctic Ocean raises questions concerning the metabolic plasticity of Antarctic notothenioids to changes in the environmental temperature. In this study, Notothenia rossii survived 90 days at 8 °C, and their condition factor level was maintained. However, their hepatosomatic (0.29×) index decreased, indicating a decrease in nutrient storage as a result of changes in the energy demands to support survival. At 8 °C, the plasma calcium, magnesium, cholesterol, and triglyceride concentrations decreased, whereas the glucose (1.91×) and albumin (1.26×) concentrations increased. The main energy substrate of the fish changed from lipids to glucose due to a marked increase in lactate dehydrogenase activity, as demonstrated by an increase in anaerobic metabolism. Moreover, malate dehydrogenase activity increased in all tissues, suggesting that fish acclimated at 8 °C exhibit enhanced gluconeogenesis. The aerobic demand increased only in the liver due to an increase (2.23×) in citrate synthase activity. Decreases in the activities of superoxide dismutase, catalase, and glutathione-S-transferase to levels that are most likely sufficient at 8 °C were observed, establishing a new physiological activity range for antioxidant defense. Our findings indicate that N. rossii has some compensatory mechanisms that enabled its long-term survival at 8 °C.

Metabolic responses in Antarctic Nototheniidae brains subjected to thermal stress.

Antarctic Nototheniidae is an attractive group for studying metabolic and physiological responses at high temperatures. The present work investigated the metabolic responses of the carbohydrate metabolism and antioxidant system to thermal stress at 8 °C (for 2-144 h) in the brains of Notothenia rossii and Notothenia coriiceps. In N. coriiceps, glycogenolysis was essential in the first hours of exposure (2 h) at 8 °C and, in addition to inhibiting glucose-6-phosphatase activity, was important for activating the pentose phosphate pathway. In N. rossii, anaerobic metabolism was reduced in the first hours of exposure (2 and 6 h) at 8 °C, followed by reduced hexokinase activity, suggesting energy regulation between neurons and astrocytes. The antioxidant system results indicated the importance of the actions of the glutathione-dependent antioxidant enzymes glutathione-S-transferase and glutathione peroxidase as well as those of catalase in N. coriiceps and the action of glutathione-S-transferase, glutathione peroxidase and glutathione reductase in N. rossii, especially during the first 12 h of thermal stress exposure. These results indicate tissue-specific patterns and species-specific responses to this stress.

Heat stress in the heart and muscle of the Antarctic fishes Notothenia rossii and Notothenia coriiceps: Carbohydrate metabolism and antioxidant defence.

Carbohydrate metabolism and the antioxidant defence system of heart and muscle of the Antarctic notothenioids Notothenia rossii and Notothenia coriiceps were evaluated in response to heat stress (8 °C) over 144 h. N. rossii heart exhibited decreased glycolysis and aerobic metabolism after up to 12 h of exposure to 8 °C, and anaerobiosis was inhibited within 24 h. However, these pathways were stimulated after 72 h at 8 °C. The consumption of glucose-6-phosphate, derived from hexokinase (HK), by glucose-6-phosphate dehydrogenase (G6PDH) decreased in N. rossii heart within 6 h at 8 °C, with a subsequent increase at 72 h. In N. rossii muscle at 8 °C, glycolysis was stimulated within 2 h by an increase in pyruvate kinase (PK), and aerobic metabolism was stimulated at 144 h, together with anaerobiosis. In N. coriiceps heart at 8 °C, glucose break down by HK decreased within 2 h and subsequently increased at 12 and 24 h. Increased glucose-6-phosphate consumption by G6PDH occurred within 6 h at 8 °C. In N. coriiceps muscle at 8 °C, glycolysis was stimulated at 2 and 6 h, with subsequent inhibition within 24 h, as indicated by HK activity. Aerobic metabolism was inhibited at 72 and 144 h at 8 °C through the inhibition of citrate synthase (CS). Heat stress caused responses were only occasional and transient in antioxidant defence system of both species in the heart and muscle, leading to increased glutathione (GSH) and decreased levels of lipoperoxidation in the heart of both species. The results obtained in this study in the heart and muscles indicate that under heat stress at 8 °C, N. rossii is more responsive than N. coriiceps with respect to carbohydrate metabolism.

Effects of heat stress on the renal and branchial carbohydrate metabolism and antioxidant system of Antarctic fish.

The objective of the present study was to assess the effect of short-term (2-144 h) heat stress (8 °C) on energy production processes and antioxidant defense systems in the kidneys and gills of Notothenia rossii and Notothenia coriiceps. Heat stress affected energy metabolism and oxidative stress parameters in a time-, tissue-, and species-dependent manner, and gills were more sensitive than kidneys to heat stress. N. rossii kidneys were able to stabilize carbohydrate metabolism after 12 h of heat stress, whereas the glycogen levels in N. coriiceps kidneys fluctuated in response to varying glucose-6-phosphatase (G6Pase) levels. The gills of N. rossii were able to stabilize their energy demand and aerobic metabolism under heat stress, whereas in the gills of N. coriiceps, changes in carbohydrate metabolic pathways depended on the exposure time: initially, anaerobiosis was activated after 6 h; the energy demand, characterized by glycogen consumption, increased after 72 h, and aerobic metabolism was activated within 144 h. With regard to the antioxidant defenses of the N. rossii kidney, it was found that levels of antioxidant enzymes were reduced during the first hours of heat stress, contributing to increased lipid peroxidation, whereas N. coriiceps kidneys did not show signs of oxidative damage. The gills of N. rossii exhibited more pronounced oxidative damage in response to heat stress than those of N. coriiceps despite the presence of increasing levels of antioxidants, likely due to tissue hypoxia.

Time course of lead induced proteomic changes in gill of the Antarctic limpet Nacella Concinna (Gastropoda: Patellidae).

The effect of increasing levels of metals from anthropogenic sources on Antarctic invertebrates is poorly understood. Here we exposed limpets (Nacella concinna) to 0, 0.12 and 0.25 µg L− 1 lead for 12, 24, 48 and 168 h. We subsequently quantified the changes in protein abundance from gill, using 2D gel electrophoresis and mass spectrometry. We identified several antioxidant proteins, including the metal binding Mn-superoxide dismutase and ferritin, increasing abundances early on. Chaperones involved in the redox-dependent maturation of proteins in the endoplasmic reticulum (ER) showed higher abundance with lead at 48 h. Lead also increased the abundance of Zn-binding carbonic anhydrase at 12 h, suggesting a challenge to acid-base balance. Metabolic proteins increased abundance at 168 h, suggesting a greater ATP demand during prolonged exposure. Changes in abundance of the small G-protein cdc42, a signaling protein modifying cytoskeleton, increased early and subsequently reversed during prolonged exposure, possibly leading to the modification of thick filament structure and function. We hypothesize that the replacement of metals initially affected antioxidant proteins and increased the production of reactive oxygen species. This disrupted the redox-sensitive maturation of proteins in the ER and caused increased ATP demand later on, accompanied by changes in cytoskeleton. SIGNIFICANCE: Proteomic analysis of gill tissue in Antarctic limpets exposed to different concentrations of lead (Pb) over a 168 h time period showed that proteomic changes vary with time. These changes included an increase in the demand of scavenging reactive oxygen species, acid-base balance and a challenge to protein homeostasis in the endoplasmic reticulum early on and subsequently an increase in energy metabolism, cellular signaling, and cytoskeletal modifications. Based on this time course, we hypothesize that the main mode of action of lead is a replacement of metal-cofactors of key enzymes involved in the scavenging of reactive oxygen species and the regulation of acid-base balance.