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The toxicological effect of cellular extract of cyanobacterium Radiocystis fernandoi strain R28 containing RR and YR microcystins was analyzed in the fish Hoplias malabaricus with emphasis on the liver structure and energetic metabolism, after short-term exposure. Fish were intraperitoneally (i.p.) injected with 100 μg of equivalent MC-LR kg −1 body mass containing in the cellular extract of R. fernandoi strain R28. Twelve and 96 h post-injection, the plasma, liver and white muscle were sampled for biochemical analyses and liver was also sampled for morphological analyses. After i.p. injection, the activity of acid phosphatase (ACP), alanine aminotransferase (ALT) and direct bilirubin increased in the plasma, while ALT and aspartate aminotransferase (AST) decreased in the liver. Glucose, lactate and pyruvate increased while protein decreased in the plasma; glycogen, pyruvate and lactate decreased in the liver; and glycogen and glucose increased in the muscle. Ammonia increased in the plasma, liver and muscle. The hepatocyte cell shape changed from polyhedral to round after cellular extract injection; there was loss of biliary canaliculus organization, but the biliary duct morphology was conserved in the liver parenchyma. In conclusion, microcystins present in the cellular extract of R. fernandoi strain R28 affect the liver structure of H. malabaricus , but the liver was able to continuously produce energy by adjusting its intermediate metabolism; glycogenolysis and gluconeogenesis maintained glucose homeostasis and energy supply.

article i nfo Omnivorous fish utilize dietary carbohydrates better than carnivorous ones due to striking morphological and physiologicaldifferencesintheirdigestivetracts.However,digestivetractmorphologyandphysiologyalsovaries among omnivorous fishes, which can lead to different dietary utilization of plant sources as well. Starch, energy, and dry matter apparent digestibility coefficient (ADC) of wheat bran, cassava residue, ground corn, and broken rice were compared between the omnivorous freshwater jundia catfish (93.9 ± 34.0 g; mean ± standard deviation) and Nile tilapia (93.7 ± 51.6 g) by using 5 g kg �1 chromic oxide as a diet marker. Starch ADCs were significantly higher in tilapia for all plant sources (92.02% to 99.74% versus 55.87% to 90.61%), except for wheat bran, which was similar to that found in jundia. Starch-richer plant sources (ground corn and broken rice) showed the lowest starch digestibility for jundia. However, ground corn provided significantly higher energy digestibility in jundia (55.35%), as opposed to broken rice (86.59%) and ground corn (71.68%), in tilapia. Dry matter ADCs ranged from 22.89% for cassava residue in jundia to 89.17% for broken rice in tilapia. Higher dry matter ADCs were registered in tilapia for all plant sources, except for wheat bran. Despite presenting lower starch and energy digestibilities, jundia catfish showed an adaptive capacity to utilize starch sources by present- ing significantly higher specific activity of amylase (58 U mg protein �1

Metabolic responses and the growth performance of pacu Piaractus mesopotamicus were evaluated in fish reared under sustained-swimming (SS) conditions at three water flow speeds of 1, 2 and 3 BL sec-1 (Body Length sec-1) or motionless water for 50 days in recirculating aquaculture systems (RAS). The intermediary metabolites: glucose, lactate, pyruvate, amino acids, ammonia, triglycerides, and free fatty acids, plus the metabolism key enzymes, LDH, MDH, GDH, and transaminases, were assayed. The fish reared at 2 BL sec-1 exhibited improved energetic metabolism. The lipid and carbohydrate oxidative processes were enhanced in fish submitted to SS. The SS shifted 51% of dietary protein toward body growth at 2 BL sec−1, and long-term exercise at 1 BL sec-1 increased the fish body mass 21%. Sustained swimming in RAS is a promising practice in the rearing of pacu, possibly lowering diet costs and contributing to environmental water quality.

Parameters of the antioxidant defense systems of Brycon amazonicus (matrinxã--a neotropical fish) exposed to phenol for 96 h plus the recovery over 1 and 2 weeks were studied in erythrocytes and liver. Hematocrit increase was observed during phenol exposure and recovery for 1 week. Total superoxide dismutases (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH) did not change during phenol exposure. Erythrocyte glucose-6-phosphate dehydrogenase (G6PDH) increased during that period while catalase (CAT) activity decreased during phenol exposure and recovery for 2 weeks. In the liver, SOD and CAT did not change, whereas GPx increased in the first week of recovery and decreased after 2 weeks. A late response was observed for G6PDH activity which increased only at the second week. Ascorbate concentration in the brain decreased during phenol exposure and increased over recovery. From our results it appears that the oxidative stress was limited in matrinxã exposed to phenol, but seemed to occur during the recovery period.

In this study we measured plasma cortisol, plasma glucose, plasma sodium and potassium, and liver and gill hsp70 levels in juvenile matrinxã (Brycon amazonicus) subjected to a 96 h exposure to phenol (0, 0.2, and 2.0 ppm), and the effect of this exposure on their ability to respond to a subsequent handling stress. Fish were sampled prior to initiation of exposure and 96 h, and at 1, 6, 12, and 24 h post-handling stress. During the 96 h exposure, plasma cortisol and glucose levels remained unchanged in all treatments. While plasma sodium levels were significantly reduced in all groups, plasma potassium levels only decreased in fish exposed to 0 and 0.2 ppm of phenol. Liver hsp70 levels decreased significantly at 96 h in fish exposed to 2.0 ppm of phenol. All groups, except fish exposed to 0.2 ppm of phenol, were able to increase plasma cortisol and glucose levels after handling stress. Fish exposed to 2.0 ppm of phenol showed decreased gill and liver hsp70 levels after the handling stress. Our data suggest that exposure to phenol may compromise the ability of matrinxã to elicit physiological responses to a subsequent stressor.

Digestive enzyme responses plus metabolic implications were studied in tambaqui (Colossoma macropomum) fed isoproteic diets containing 28% crude protein, 3300 kcal of gross energy/kg and different amounts of cornstarch (30, 40 and 50%). Amylase, maltase, acid protease, trypsin and chymotrypsin from the alimentary tract were assayed. Plasma, liver and white muscle metabolites were gauged to profile metabolism of the fish. The alimentary tract of tambaqui is compartmentalized morphologically and enzymatically. Amylase was present through the gut; acid protease was detected in stomach; maltase, trypsin and chymotrypsin were found in pyloric caeca and proximal and distal intestine sections. Increase of cornstarch levels from 40 to 50% in the diet resulted in an increase in amylase and maltase. Trypsin and chymotrypsin were unresponsive to starch levels. Acid protease follows the protein/carbohydrate ratio decrease. The increase of dietary cornstarch resulted in liver glycogenesis and the increase in plasma triglycerides is suggestive of lipogenesis. Digestive biochemical responses of tambaqui correlated with changes of feeding plus the analyses of metabolic profile are assumed as a tool for optimizing diet formulation and are a clue to other feeding optimizations for freshwater tropical species.

This manuscript reports changes in key enzymes and metabolites related to protein metabolism and nitrogen excretion in the liver of juveniles jundiá (Rhamdia quelen) fed on isocaloric diets containing 20%, 27%, 34% and 41% of crude protein. The hepatic activity of alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), glutamate dehydrogenase (GDH), and arginase (ARG) increased with the content of protein in the diet, and the ratios among the aminotransferases and GDH allowed evaluating metabolic preference. The concentration of free amino acids, ammonia and urea also rose with the dietary protein content. Increase of plasma urea and ammonia was the resultant effect of over amino acids catabolism as consequence of dietary protein surplus. Since the increase of protein in the diets resulted in weight gain, the rise in the hepatic activity of protein-metabolising enzymes in the fish fed high protein diets denoted effective use of dietary amino acids for growth and as a substrate for gluconeogenesis. Analysis of changes on metabolite levels and key enzyme activities in amino acid metabolism is proposed as a tool for assessing the proper balance of diet macronutrients.

Effects of increasing dietary protein were studied in matrinxa, (Brycon cephalus), an omnivorous teleost from the Amazon Basin in Brazil. Four isocaloric diets were formulated to contain 20%, 27%, 34% and 41% of crude protein (CP). Plasma glucose levels were significantly increased while triacylglycerols were significantly reduced at 41% of CP. Free fatty acids were significantly reduced at each level of rising CP. Plasma amino acids and ammonium followed the dietary CP increase. Liver glycogen and amino acids were reduced; liver glucose and lactate were constant, and ammonium increased with the CP in diets. Muscle glycogen and pyruvate decreased, protein did not change, while lactate and free amino acids increased. Kidney glycogen proportionally rose with the increase of CP from 20-41%. Pyruvate and lactate augmented irregularly from 20-41% CP. A gluconeogenic profile was observed in the kidney; the liver worked as regulator of body glucose. Increase of dietary CP and decrease of carbohydrates pushed muscle and liver catabolism of fat and sugar stores to satisfy energetical demands. CP contents above 34% were not recommended for B. cephalus, if the carbohydrate does not compensate the metabolical demands.

Hemolytic anemia accompanied by changes in the immunology system is one of the sulfide intoxication harmful effects on Hoplosternum littorale. Hematological parameters are considered as effective indicators of stress caused by this hydrogen sulfide. During sulfide exposure, H. littorale neither alters the methemoglobin concentration nor forms sulfhemoglobin in the presence of high levels of dissolved sulfide in the water. Cytochrome c oxidase shows little activity in the gills and blood of H. littorale when exposed to sulfide. Alternative metabolic routes are suggested through which the accumulation of pyruvate leads to the formation of an end product other than lactate.

The silver catfish (Rhamdia quelen) is a teleost native to South America. It can resist cold winters and grow quickly in the summer and is an important species for aquaculture in both temperate and subtropical climates. The effect of different water temperatures (15, 23 and 31 °C) on hematological and metabolic parameters in blood, liver and white muscle was investigated in this species following chronic (21 days) and acute (12 h) exposure. In both experiments, hematocrit, hemoglobin and cortisol were unchanged, but plasma glucose levels increased at a temperature of 31 °C and decreased at 15 °C, when compared to control. Fatty acid levels decreased at 31 °C, and triacylglycerol levels increased at 15 °C in the 21-day experiment compared to control. One of the most interesting results of this study was the decrease of total protein in the liver and white muscle of fish exposed to 31 °C for 21 days. Following 21 days, liver glycogen levels decreased at 15 and 31 °C and glucose and lactate contents increased at 31 °C compared to control. In white muscle, glycogen and glucose levels increased at 15 and 31 °C, while lactate decreased at 31 °C. No alterations were observed in the liver or white muscle of fish exposed to the different temperature for 12 h. Taken together, these results suggest that temperature and time of exposure influence metabolic parameters in the plasma, liver and white muscle of silver catfish.

The neotropical freshwater fish matrinxa, Brycon cephalus (Gunther, 1869), was exposed to 1/3 of 96h-LC50 of Folidol 600 (methyl parathion) for 96 h and allowed to recover for 24, 48, 96, and 192 h. Acetylcholinesterase (AChE), alanine aminotransferase (ALAT), aspartate aminotransferase (AAT), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), and glutamate dehydrogenase (GDH) were assayed in brain, liver, muscle, and plasma. Plasma and brain AChE activities were strongly inhibited (64% and 87%, respectively), remaining low in the brain during recovery. Liver ALAT decreased 59.4% and plasma ALAT increased 94.2%. This response, associated with GDH reduction during the recovery period, was attributed to impairment of amino acid metabolism and to liver damage. The increase of heart and plasma AAT suggested tissue injury. Liver and plasma LDH and MDH did not change during the exposure to Folidol, but liver MDH decreased 34% during recovery, indicating a failure of oxidative metabolism in such tissue. Hepatic glycogen and glucose levels decreased 80.4 and 55%, respectively, followed by glucose mobilization to the plasma. The hepatic and muscular profile of lactate after recovery suggested gluconeogenesis without tissue hypoxia. These results revealed that methyl parathion affects the intermediary metabolism of matrinxa and that the assayed enzymes can work as good biomarkers of organophosphorus pesticide contamination.

Digestive enzyme responsiveness to feeding and associated adjustments of metabolism can be used to derive nutritionally effective diet formulations. Juvenile pintado (Pseudoplatystoma corruscans) were fed different diets. After feeding, fish were killed and blood, liver and white muscle were collected to evaluate metabolites. Stomach along with anterior, middle and posterior intestine were sampled for enzyme analysis. Non-specific protease, trypsin, chymotrypsin, amylase and lipase were assayed. Crude protein (CP) did not induce proteolytic activity; highest protease activities were observed in the stomach. Amylase was higher in the stomach in fish feeding on diets containing 13-25% starch. Lipase activity was observed along the gastrointestinal tract, with the highest activities observed in the middle section. The metabolic profile of white muscle was not affected by CP. In contrast, some plasma and liver metabolites were altered concomitant with changes in the digestive enzymes. Amino acid catabolism was increased. Digestion in pintado was responsive to cornstarch, reflected in intermediary metabolism; proteolytic activities of the digestive tract seem to be sufficient to deal with large amounts of dietary protein. As a result, we are able to recommend a balance between protein and energetic compounds, such as lipids and carbohydrates, in the diet to optimize fish growth.

Juvenile tambaqui, Colossoma macropomum, were exposed to sulfide and hypoxia for 12, 24, 48 and 96 h. Hemoglobin concentrations, red blood cell counts, and mean cell hemoglobin, were higher at 12 h in fish exposed to hypoxia. However, control fish and those exposed to sulfide and hypoxia had lower red blood cell count, hemoglobin concentration and hematocrit at 96 h. Methemoglobin was higher than in the controls, probably due to the hypoxemia induced by these stressors. Sulfhemoglobin was not detected in significant amounts in the blood of fish exposed to sulfide (in vivo), yet hemoglobin converted into sulfhemoglobin at 1-15 mM sulfide in vitro. Anaerobic metabolism seemed to be an important mechanism for adapting to sulfide exposure and blood pH returned to control values after 24 h of sulfide, preventing acidosis. The high sulfide tolerance in tambaqui is associated with its high tolerance to hypoxia.

Eight species of trypanosomatid flagellates, Trypanosoma cruzi, T. mega, T. conorhini, Leishmania donovani, L. braziliensis, Leptomonas seymouri, L. collosoma, and L. samueli, were examined for the presence of enzymes of the arginine-ornithine metabolism. Arginase was found in species of the genera Leishmania and Leptomonas. Citrulline hydrolase was found only in species of Leptomonas. Trypanosoma spp. did not present any of the mentioned enzymes. Ornithine carbamoyltransferase and argininosuccinate lyase were found only in Leptomonas samueli, which also possessed arginine deiminase. With the sole exception of L. samueli the other species seem to present a uniform enzyme constitution, peculiar to their genera and different from the enzyme patterns of other genera of trypanosomatids already known. The potential usefulness of these findings for taxonomical purposes is discussed.