Cândida Toni

Assessment of oxidative stress in Rhamdia quelen exposed to agrichemicals.

Due to the proximity of crop and fish culture areas, some agrichemicals that could be harmful for fish could enter into fishponds by different ways, such as by leaching through rain. Rhamdia quelen (Teleostei) were exposed to sublethal concentrations of methyl parathion (MP), a glyphosate based herbicide (Gly), and tebuconazole (Teb). The liver of R. quelen exposed to MP and Teb showed enhanced levels of thiobarbituric acid reactive substances (TBARS), higher than in the control fish (56% and 59%, respectively). In contrast, Gly did not alter the TBARS generation. The protein carbonyl content increased only in fish exposed to Teb. Fish exposed to the three agrichemicals showed a significant decrease of catalase activity (52%, 48%, and 67%, respectively) and increased glutathione-S-transferase (57%, 46%, and 160%, respectively) activity. Fish exposed to MP, Gly, and Teb showed higher reduced glutathione (151%, 472%, and 130%, respectively, when compared with the control levels) and ascorbic acid concentrations (121%, 102%, and 184%, respectively),while the non-protein thiol content increased only in R. quelen exposed to tebuconazole. Fish exposed to MP and Teb showed several pathological changes in the liver, including hepatocyte degeneration and bile stagnation. The present work reports for the first time the toxicity of the pesticide MP and the fungicide Teb in R. quelen, and as in other works, suggests the relatively lower liver toxicity of Gly for fish. The data presented herein demonstrate that sublethal concentrations of MP and Teb cause changes in oxidative stress parameters as well as hepatic cell injuries in R. quelen, and that these parameters have the potential to be developed as bioindicators of exposure to these agrichemicals.

Toxicological responses of Cyprinus carpio after exposure to a commercial herbicide containing imazethapyr and imazapic

Cyprinus carpio was exposed to imazethapyr and imazapic at laboratory and at field conditions. The laboratory experiment was carried out for 7 days and at rice field for 7, 30 and 90 days. Oxidative stress parameters and antioxidant profile were studied as well as metabolic parameters. After 7 days, brain AChE activity increases in laboratory and field, but in muscle, reduction was observed only in laboratory. At the same period, brain and muscle TBARS and liver CAT increase in the laboratory. Metabolic parameters showed changes in both conditions and exposure periods. After 30 days in rice field, brain AChE activity decreases and in muscle it was enhanced. After 90 days in field, only muscle AChE activity was reduced. The disorders in oxidative stress parameters and metabolism remained, indicating mainly a protein catabolism. This study pointed out short- and long-term effects of rice herbicides at environmentally relevant concentrations on toxicological parameters in tissues of C. carpio.

Oxidative stress biomarkers in Cyprinus carpio exposed to commercial herbicide bispyribac-sodium.

Cyprinus carpio were exposed under field conditions to 20.87 µg l−1 of commercial herbicide bispyribac‐sodium (Nominee®, SC), during 7, 21 and 72 days. Enzymatic parameters such as catalase (CAT), glutathione S‐transferase (GST) and acetylcholinesterase (AChE) activities, as well as thiobarbituric acid‐reactive substances (TBARS) and protein carbonyl contents were studied in different tissues. After 7 days of exposure, GST activity decreased. At the same period, brain AChE activity increased, but a reduction of activity was observed in muscle tissue. Brain TBARS levels increased at 7 days. After 21 days of exposure liver CAT levels and muscle AChE activities decreased. In the same period, liver protein carbonyl and muscle TBARS increased. After 72 days of exposure in the field, AChE activity was reduced in both brain and muscle. Protein carbonyl contents in liver and brain TBARS levels increased. Muscle AChE activity, TBARS and protein carbonyl can be used as biomarkers of exposure to the herbicide bispyribac‐sodium. This study demonstrates effects of exposure to bispyribac‐sodium under rice field conditions on oxidative stress parameters in tissues of Cyprinus carpio. Copyright

Assessment of oxidative stress and metabolic changes in common carp (Cyprinus carpio) acutely exposed to different concentrations of the fungicide tebuconazole.

The aim of this research is to determine the lethal concentration (LC(50-96 h)) of fungicide tebuconazole for common carp (Cyprinus carpio) and to investigate the occurrence of oxidative stress and metabolic changes after acute exposure to different concentrations of tebuconazole. For this purpose, several parameters were assessed as whole-body levels of thiobarbituric acid-reactive substance (TBARS), protein carbonyl, catalase, superoxide dismutase, glutathione S-transferase, nonprotein thiols, ascorbic acid, glycogen, glucose, lactate, protein, amino acids and ammonia in tebuconazole-exposed fish. The calculated LC(50-96 h) was 2.37 mg L(-1). Fish exhibited significant increase in TBARS levels in all concentrations used while the enzymatic and nonenzymatic antioxidants were decreased. Among the metabolic parameters, glycogen and glucose increased at 1.19 mg L(-1) and protein levels decreased at 1.78 and 2.37 mg L(-1). In conclusion, the fish health was adversely affected by exposure to tebuconazole, and those changes can compromise animal survival in the natural environment. The results indicate that the some of the parameters measured like a possible biomarkers of exposure to tebuconazole for this species of fish.

The effects of ammonia and water hardness on the hormonal, osmoregulatory and metabolic responses of the freshwater silver catfish Rhamdia quelen

The aim of the present study was to assess the effects of ammonia and water hardness on endocrine, osmoregulatory and metabolic parameters in silver catfish (Rhamdia quelen). The specimens (60-120g) were subjected to six treatments in triplicate, combining three levels of un-ionized ammonia (NH3) (0.020±0.008mg/L [1.17±0.47μM], 0.180±0.020mg/L [10.57±1.17μM] and 0.500±0.007mg/L [29.36±0.41μM]) and two levels of water hardness (normal: 25mgCaCO3/L and high: 120mgCaCO3/L), and sampled after two exposure times (1 and 5 days post-transfer). Plasma cortisol, metabolites, osmolality and ionic values were determined concomitantly with the mRNA expression levels of different adenohypophyseal hormones (growth hormone, GH; prolactin, PRL; and somatolactin, SL). Previously, full-length PRL and SL as well as β-actin cDNAs from R. quelen were cloned. Exposure to high NH3 levels enhanced plasma cortisol levels in fish held under normal water hardness conditions but not in those kept at the high hardness value. The increase in water hardness did not alter plasma metabolites, whereas it modulated the osmolality and ion changes induced by high NH3 levels. However, this hardness increase did not lead to the decreased GH expression that was observed 5 days after exposure to 0.18mg/L NH3 in fish held at the normal water hardness level, whereas PRL expression was enhanced after one day of exposure under the increased hardness conditions. Additionally, SL expression decreased in specimens exposed for 5 days to 0.18mg/L NH3 and maintained at the high water hardness level. The results showed that increasing water hardness attenuated the hormonal parameters evaluated in R. quelen specimens exposed to high NH3 levels, although plasma metabolism do not appear to suffer major changes.

Sedative effect of 2-phenoxyethanol and essential oil of Lippia alba on stress response in gilthead sea bream (Sparus aurata)

The anesthetic efficacy of the essential oil of Lippia alba (EOLA) in Sparus aurata was evaluated by induction and recovery times of anesthesia. After, specimens were exposed to anesthetics low concentrations for 4h, under nonstress or stress conditions. Range 100-300 μL L(-1) EOLA induced anesthesia. Plasmatic cortisol, glucose, lactate, and osmolality enhanced after EOLA exposure in the undisturbed (UF) and stressed fish (SF). Lower corticotropin-releasing hormone binding-protein expression occurred in SF/EOLA compared with 2-PHE/stress combination or to EOLA/undisturbed conditions. Stress processes reduced prolactin (PRL) expression in the control fish, while UF exhibited reduced PRL levels after exposure to both anesthetics. Proopiomelanocortin (POMCa) mRNA was higher after 2-PHE exposure in SF compared to control; POMCb expression was higher in SF/EOLA in contrast to control and UF/EOLA conditions. Thus, EOLA was an effective anesthetic, but it was unable to prevent a stress response in S. aurata; while 2-PHE is advisable to sedate S. aurata without causing stress, but it was not effective at preventing a stress response in the present work.

Exposure to tebuconazol in rice field and laboratory conditions induces oxidative stress in carp (Cyprinus carpio).

Pesticides can have an effect on the biochemical and physiological functions of living organisms. The changes seen in fish and their response to pesticides can be used as an example for vertebrate toxicity. In this study, carp fish (Cyprinus carpio) were exposed to different concentrations of tebuconazol fungicide, by rice field (31.95 μg/L) and laboratory (33.47 and 36.23 μg/L) conditional testing, during a 7 day period. Parameters such thiobarbituric acid-reactive substance levels (TBARS), protein carbonyl, catalase, glutathione S-transferase and acetylcholinesterase activities were studied, using the liver, brain and white muscle of the fish. The field experiment showed that the TBARS levels were increased in all the analyzed tissues. Similarly, the protein carbonyl of the liver and the brain AChE activity increased after 7 days. The laboratory experiment demonstrated that the TBARS levels in the liver were increased in both of the concentration tests. TBARS levels in the muscle increased only by the lowest test concentration. On the other hand, the protein carbonyl was increased only by the highest concentration. The results indicate that the tebuconazol exposure from the field and laboratory conditions directly affected the health of the fish, showing the occurrence of oxidative stress.

Alterations in carbohydrate and protein metabolism in silver catfish (Rhamdia quelen) exposed to cadmium.

Changes in carbohydrate and protein metabolism were studied in silver catfish Rhamdia quelen exposed to cadmium (0; 0.236 or 0.414 mg/L) during 7 and 14 days. After exposure time the fish were exposed to recovery period (water without cadmium), during 7 and 14 days. Different alterations in the metabolic parameters were observed such as an increase in lactate, protein, amino acid and ammonia levels as well as a reduction in glucose values after the exposure periods in liver. In muscle, glycogen and glucose values enhanced after cadmium exposure at both concentrations for 7 days; however, at 0.414 mg/L cadmium, protein levels decreased while amino acids and ammonia levels enhanced. An increase in the lactate values was found in plasma after 7 days of exposure and a reduction in the lactate, glucose and protein levels occurred after 14 days of exposure. Results indicated that the metabolic alterations after cadmium exposure were dependent on the tissue type and exposure time. Cadmium exposure for 14 days and recovery period also of 14 days seem to be less harmful to the liver and muscle. However, even after recovering from some changes, fish health may be affected making them more sensitive to some environmental changes.

Comparative study on effects of dietary with diphenyl diselenide on oxidative stress in carp (Cyprinus carpio) and silver catfish (Rhamdia sp.) exposed to herbicide clomazone

The study investigated the capacity of diphenyl diselenide [(PhSe)2] (3.0mg/kg), on reduce the oxidative damage in liver, gills and muscle of carp and silver catfish exposed to clomazone (192h). Silver catfish exposed to clomazone showed increased thiobarbituric acid-reactive substance (TBARS) in liver and muscle and protein carbonyl in liver and gills. Furthermore, clomazone in silver catfish decrease non-protein thiols (NPSH) in liver and gills and glutathione peroxidase and ascorbic acid in liver. (PhSe)2 reversed the effects caused by clomazone in silver catfish, preventing increases in TBARS and protein carbonyl. Moreover, NPSH and ascorbic acid were increased by values near control. The results suggest that (PhSe)2 attenuated the oxidative damage induced by clomazone in silver catfish. The clomazone no caused an apparent situation of oxidative stress in carp, showing that this species is more resistant to this toxicant. Altogether, the containing (PhSe)2 diet helps fish to increase antioxidants defenses.

Oxidative stress in carp exposed to quinclorac herbicide under rice field condition.

This study evaluated parameters of oxidative stress and antioxidant profile in fish after herbicide exposure. Cyprinus carpio were exposed to quinclorac (initial concentration 344.60μg/L) for 7, 30, and 90 days under rice field condition. Thiobarbituric acid-reactive substances (TBARS) were evaluated in brain, liver and muscle tissues, and protein carbonyl in liver. Enzymatic parameters such as catalase (CAT) and gluthatione S-transferase (GST) activities also were studied in liver. TBARS levels fluctuated in the brain showing increase in 7 days and decrease in 30 days, while in liver it was observed increase in 7 and 30 days, as well as in muscle after 30 and 90 days. The protein carbonyl was also increased after 30 and 90 days of herbicide exposure. CAT and GST activities were decreased after 30 and 90 days, respectively. The alterations observed suggest that a commercial formulation containing quinclorac causes oxidative damage in different tissues of carp after a long time of exposure. This study pointed out the importance of quinclorac toxicity considering the concentration used in rice fields.