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.

Yarrowia lipolytica biomass-a potential additive to boost metabolic and physiological responses of Nile tilapia.

We studied the effects of Yarrowia lipolytica biomass on digestive enzymes, blood biochemical profile, energy metabolism enzymes, and proximate meat composition of Nile tilapias. The experiment was entirely randomized with four replications. The animals (n = 20 per repetition) were fed with 0%, 3%, 5%, and 7% of biomass for 40 days and then blood and liver were analyzed. There was an increase in the activities of chymotrypsin (5, 7% groups), trypsin (3, 5% groups), and sucrase (7% group) compared to the respective control groups. On the other hand, maltase activity was significantly reduced for all yeast biomass treatments, while the supplementation did not influence lipase and amylase activities. Moreover, the blood triacylglycerol concentrations were increased in the 7% group, while any treatment modified blood total cholesterol, glycemia, and hepatic glycogen content. Y. lipolytica biomass promoted significant increases in meat protein and lipid contents without changes in moisture and ash parameters. Furthermore, Y. lipolytica biomass promoted increases in hexokinase (3% group), phosphofructokinase (5, 7% groups), glucose-6-phosphate dehydrogenase (5% group), citrate synthase (3% group), aspartate aminotransferase and alanine aminotransferase (3% group), and glutamate dehydrogenase (3, 5% groups) compared to the respective control groups. At the same time, no changes were observed in the activity of glucose-6-phosphatase. Y. lipolytica biomass supplementation in tilapias' diet can modulate the digestive system and improve nutrient disponibility to the cells. Moreover, the changes in the metabolic profile and in energy metabolism can be correlated with the improvement of meat composition. Therefore, the Y. lipolytica biomass has a great potential to be used as a feed ingredient for Nile tilapias.

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.

Effects of heat shock on energy metabolism and antioxidant defence in a tropical fish species Psalidodon bifasciatus.

Predictions about global warming have raised interest in assessing whether ectothermic organisms will be able to adapt to these changes. Understanding the physiological mechanisms and metabolic adjustment capacity of fish subjected to heat stress can provide subsidies that may contribute to decision-making in relation to ecosystems and organisms subjected to global climate change. This study investigated the antioxidant defence system and energy metabolism of carbohydrate and protein responses in the gill, liver and kidney tissues of Psalidodon bifasciatus (Garavello & Sampaio 2010), a Brazilian freshwater fish used in aquaculture and in biological studies, following exposure to heat shock at 31°C for 2, 6, 12, 24 and 48 h. The fish presented signs of stress in all tissues tested, as evidenced by increased lipid peroxidation concentration at 2 h and phosphofructokinase, hexokinase and malate dehydrogenase activity at 48 h in the gills; increased glutathione-S-transferase activity at 12 h, citrate synthase activity at 24 h and concentration of reduced glutathione (GSH) concentration at 12 and 48 h in the liver; and through increased activity of superoxide dismutase at 48 h, glutathione reductase at 24 h, glucose-6-phosphate dehydrogenase at 48 h and concentration of GSH at 24 h in the kidney. In the kidneys, changes in the antioxidant system were more prominent, whereas in the gills, there were greater changes in the carbohydrate metabolism. These results indicated the importance of glycolysis and aerobic metabolism in the gills, aerobic metabolism in the liver and pentose-phosphate pathway in the kidneys during homeostasis. The biomarker response was tissue specific, with the greatest number of biomarkers altered in the gills, followed by those in the kidneys and liver.

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.

Immunomodulatory effects of Yarrowia lipolytica as a food additive in the diet of Nile tilapia.

The use of yeasts as a dietary additive for fish can act as a source of nutrients and as an immunostimulant. This work aimed to evaluate the effects of the fermented biomass of the yeast Yarrowia lipolytica as a food additive on zootechnical and hematological parameters, and on immune response in the plasma and kidney of Nile Tilapia (Oreochromis niloticus). After supplementation with 3, 5, and 7% of the yeast biomass for 35 days, the blood and tissues of the animals of each experimental group were collected for analysis. The addition of this biomass in the feed promoted an improvement of zootechnical parameters in tilapia. There was also a rise in the number of neutrophils (groups with 3, 5, and 7%) and monocytes (group 3, 5, and 7%) compared with the control group. Moreover, there was an increase in the levels of lysozyme, myeloperoxidase, and nitrite/nitrate content in the blood of animals fed with yeast biomass. On the other hand, there were no observed alterations in survival and hematological parameters of animals fed with yeast biomass. In the analysis of the kidney, the addition of biomass in feed promoted an increase in levels of myeloperoxidase (group with 3%) but did not alter the levels of lysozyme and nitrite/nitrate content. In conclusion, this study demonstrated that Y. lipolytica had growth and immunostimulatory effects on Nile tilapia. These findings strongly suggest the potential application of a Y. lipolytica-based immunostimulant for tilapia aquaculture.

Assessment of antioxidant system, cholinesterase activity and histopathology in Lithobates catesbeianus tadpoles exposed to water from an urban stream.

Anthropogenic activities promote changes in community structure and decrease the species abundance of amphibians. The aim of this study was to assess potential alterations in the antioxidant system and cholinesterase activity, histopathological and oxidative damage in Lithobates catesbeianus tadpoles exposed to water from the Cascavel River, in Southern Brazil. Water samples (140 L each) were collected from the headwater, urban and rural areas of the river. Tadpoles were acclimated for seven days. After acclimatization tadpoles were reared in water from the river, except for the control aquarium. After seven days, a portion of the liver was removed and prepared for cholinesterase (ChE), superoxide dismutase (SOD), catalase (CAT) and lipid peroxidation (LPO) analysis; another part of the tissue was prepared for histological examination. An elevation of CAT activity was observed for water from both urban and rural environments. A decrease in LPO reaction was detected, mainly among the tadpoles exposed to water from the rural area. These alternations can cause delay the metamorphosis and lead to metabolic dysfunction, interfering in survival capacity and diminishing, not only individual fitness, but that of the whole population.

Evaluation of the antioxidant system and neurotoxic effects observed in Rhamdia branneri (Teleostei: Heptapteridae) sampled from streams of the lower Iguazu River basin.

The use of multiple biomarkers has been shown to be an efficient method for evaluating environmental contamination. In this work, we evaluate neurotoxic effects and the antioxidant system responses of the R. branneri collected in two streams of lower Iguazu River basin, relating them with different percentage of vegetation coverture, presence of pesticides and fall and winter seasons. The biological samples were collected in March and August of 2015, from two streams that belong to the lower Iguazu River basin (Brazil): the Manoel Gomes River and the Arquimedes Stream. Soil analyses were performed, and the results showed the presence of the following organophosphates in the Manoel Gomes River and the Arquimedes Stream: disulfoton, methyl parathion, and ronnel. The present study detected inhibition of cholinesterase activity in the brain and muscle of fish samples during the fall from the Manoel Gomes River and the Arquimedes Stream. In the Manoel Gomes River, elevated lipoperoxidation was also observed during the fall. It was observed that the increase or decrease of biomarkers was related to temporal variation and, possibly, to the exposure of animals to agrochemicals. Although the Manoel Gomes River and the Arquimedes Stream are located in regions with large areas of vegetation, the soil analyses show that agrochemical residues are able to reach these locations, which suggests that the fauna are in contact with oxidant and anti-cholinesterase agents during the fall, in addition to respond differently during each season.

Assessment of the oxidative and neurotoxic effects of glyphosate pesticide on the larvae of Rhamdia quelen fish.

The aim of this work was to investigate the effects of glyphosate on the antioxidant system, as well as the neurotoxic effects on the larvae of Rhamdia quelen. A completely randomized design was implemented with the eggs of silver catfish distributed in 48 containers with 300 mL of water, which were subdivided randomly into two groups: control and treated with 6.5 mg L of glyphosate. These groups were evaluated at four time points (12 h, 24 h, 48 h, and 72 h), each with six replications. The survival rate of eggs/larvae (%) was evaluated, and samples were collected for antioxidant system analysis (catalase - CAT, glutathione transferase - GST, glutathione reductase - GR, and lipoperoxidation - LPO), and neurotoxic evaluation (cholinesterase - ChE). Throughout the 72 h of experimentation, there was a higher survival rate among the animals treated with glyphosate. The highest value of integrated biomarkers response (IBR = 1.26) was at 12 h, presenting induction of the cholinesterase (ChE) enzyme and GR. At 24 h, the value of IBR was -2.56, with inhibition of ChE and induction of GR. At 48 h, the value was -0.76, with induction of LPO. The lowest value of IBR was at 72 h (-4.65), with induction of GST and inhibition of all other biomarkers. Finally, it was possible to detect an acute effect of glyphosate throughout the early development of R. quelen, with a decrease in the antioxidant system control and neurotoxic effects.

Metabolism of the mesoionic compound (MI-D) by mouse liver microsome, detection of its metabolite in vivo, and acute toxicity in mice.

The mesoionic derivative 4-phenyl-5-[4-nitrocinnamoyl]-1,3,4-thiadiazolyl-2-phenylamine chloride (MI-D) has antitumoral and anti-inflammatory effects. In this study, we present aspects of its metabolism and toxicity in mice. MI-D was metabolized in vitro by liver microsome, generating a main product with a much shorter retention time than MI-D in high-performance liquid chromatography (HPLC) analysis but with a spectrum similar to that of the original molecule. Mass spectrometry with electrospray ionization in positive mode analysis of the purified compound by HPLC indicated that the product of metabolism has four additional hydroxyl groups (m/z = 465) compared with MI-D (m/z = 401). The HPLC analyses of plasma and urine samples from mice treated with MI-D showed the presence of the metabolite product. The kinetic parameters K(m) (19.5 +/- 4.5 microM) and V(max) [1.5 +/- 0.4 units of fluorescence/(100 microg of microsomal protein/mL/s)] were estimated, confirming the metabolism of MI-D and indicating that the reaction follows Michaelis-Menten kinetics. Acute toxicity was established on the basis of an estimation of mean lethal dose (LD-50; 181.2 mg/kg) and histopathological analysis of animals that survived the LD-50 test. Abdominal adhesions, inflammatory foci, and formation of granulomas were observed. Altogether, the results contribute to the advancement of research in support of MI-D as a future chemotherapeutic drug.

The inhibition of lipoperoxidation by mesoionic compound MI-D: a relationship with its uncoupling effect and scavenging activity.

Important biological activities have been described for mesoionic compounds. We previously reported that MI-D (4-phenyl-5-(4-nitro-cinnamoyl)-1,3,4-thiadiazolium-2-phenylamine chloride) inhibited the respiratory chain, collapsed the transmembrane potential, and stimulated ATPase activity in intact rat liver mitochondria. It is known that drugs that affect mitochondrial membrane potential may facilitate the induction of cell death by apoptosis. Mitochondria have also a central role in the generation of reactive oxygen species, therefore it would be important to investigate how MI-D could affect processes related to oxidative stress. In this work, we evaluated the effects of MI-D on the lipoperoxidation and its ability to scavenge free radicals. Interestingly, it was observed that MI-D promoted a strong inhibition of the lipoperoxidation induced by Fe(3+)-ADP/2-oxoglutarate in isolated mitochondria (95%+/-0.27 at the highest concentration of 80 nmol mg(-1) protein) in a dose-dependent manner. However, at the same concentration its effect was less intense (22%+/-3.46) when the lipoperoxidation was initiated by peroxyl radicals generated from the azocompound AAPH. Lipid peroxidation in both coupled and uncoupled submitochondrial particles initiated with Fe(2+)/NADH was also inhibited by MI-D. The inhibition was about four times greater in coupled particles (approximately 34% at 80 nmol mg(-1) protein) in relation to uncoupled. MI-D inhibited the soybean phosphatidylcholine liposomes lipoperoxidation in a dose-dependent manner (5-80 microM) regardless of the radical being generated in lipid or aqueous phase. The mesoionic compound showed ability of scavenging superoxide radical (7, 11 and 31% for 25, 38 and 80 microM, respectively). Our results strongly suggest that the inhibition of lipoperoxidation promoted by MI-D is due to its scavenger action and to its previously described uncoupling effect.

Population structure and somatic indexes of Hypostomus cf. ancistroides (Siluriformes, Loricariidae) collected from the Bonito river, Ivaí river basin, Turvo, Paraná

This study aimed to provide information about the population structure and somatic index of Hypostomus cf. ancistroides collected from the Bonito river, located in the Ivaí river basin. The length-weight relationship was isometric for both the sexes. The length structure analysis showed that the larger individuals (from 18.1cm to 27.0cm in length) predominated, and the lowest abundances occurred at the size extremes (9.1-12cm and 27.1-30.0cm). The reproduction period occured between October and January for the females and between November and January for the males. The liver somatic index cannot be used as an indicator of the reproduction period in either of the sexes, due to no correlation between the liver somatic index and the gonad somatic index. The gonad weight exerted no influence on the monthly mean condition factor and the correlation between the condition factor and gonad somatic index was high. The condition factor could be an indicator of the reproduction period of this species.

Este estudo teve o objetivo de fornecer informações sobre a estrutura populacional e os índices somáticos de Hipostomus cf. ancistroides coletados no rio Bonito localizado na bacia do rio Ivaí. A relação comprimento-peso, para machos e para fêmeas, foi isométrica. A análise da estrutura em comprimento mostrou que há um predomínio de indivíduos maiores (entre 18.1 a 27.0 cm) sendo que as menores abundâncias ocorreram nas classes de comprimentos extremas (9.1 a 12.0 cm e 27.1 a 30.0 cm). O período reprodutivo em fêmeas ocorreu entre os meses de outubro a janeiro. Os machos possuem um período reprodutivo entre novembro e janeiro. O índice hepatossomático, para ambos os sexos, não pode ser utilizado como indicador do período reprodutivo, devido à não correlação entre o índice hepatossomático e o índice gonadossomático. O peso das gônadas não influenciou o valor médio do fator de condição e foi observada uma alta correlação entre o fator de condição e o índice gonadossomático. O fator de condição pode ser um indicador do período de reprodução desta espécie.

Blood parameter analysis and morphological alterations as biomarkers on the health of Hoplias malabaricus and Geophagus brasiliensis

Este trabalho teve como objetivo avaliar a influência do ambiente sobre a higidez dos peixes. Animais, das espécies Hoplias malabaricus e Geophagus brasiliensis foram coletados em três ambientes distintos, sendo ambiente I região urbana e ambientes II e III em região rural. Foram realizadas análises do número total de eritrócitos por microlitro de sangue, microhematócrito, taxa de hemoglobina, porcentagem de leucócito e contagem diferencial de leucócitos em extensão sanguínea. Calcularam-se os índices hematimétricos absolutos: volume corpuscular médio e concentração de hemoglobina corpuscular média. Para análises das alterações morfológicas, brânquias, fígado, gônadas e rim seguiram processamento histológico de rotina. Foram observados índices hematológicos ligeiramente menores em indivíduos coletados no ambiente III em relação aos animais coletados no ambiente I. As análises histológicas de brânquias, fígado e gônadas das espécies G. brasiliensis e H. malabaricus demonstraram alterações morfológicas em porcentagens baixas. Foram identificas alterações internas nos rins, sendo representadas por degeneração e cristalização no interior dos túbulos renais. Em 92% dos indivíduos de G. brasiliensis, coletados no ambiente I, foi observado cristalizações no interior de túbulos. Sugere-se que tais alterações estejam relacionadas com a baixa circulação de água no local.

Histopathology of the fish Corydoras paleatus contaminated with sublethal levels of organophosphorus in water and food.

The effects of contamination, through water or food, of a sublethal dose of the organophosphate methyl parathion were analyzed in tissues that are responsible for absorption (gills, intestine) and metabolism (liver), in the freshwater fish Corydoras paleatus. In gill respiratory lamellae, epithelial hyperplasia, edema, and detachment occurred, diminishing sooner after contamination by food than after contamination through water. In the intestine, lipoid vacuolization of enterocytes, apical cytoplasm, and an increase in goblet cell activity occurred mainly after ingestion of contaminated food. The liver exhibited cloudy swelling, bile stagnation, focal necrosis, atrophy, and vacuolization after contamination through both absorption routes, the highest degeneration being between T(8) and T(24). Metabolic processes that depend on liver function were equally impaired by the two routes of contamination, but secondary effects vary with gill and intestine pathologies as a consequence of water and food contamination, respectively. Therefore, a "safe" sublethal dose of methyl parathion causes serious health problems in C. paleatus.