Daiane Ferreira

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.

Diazepam and Fluoxetine Decrease the Stress Response in Zebrafish

The presence of pharmaceutical products in the aquatic environment has been reported in several studies. However, the impact of these drugs on living organisms is still uncharacterized. Here, we investigated the effects of acute exposure to either diazepam or fluoxetine on the stress response in Danio rerio. We showed that diazepam and fluoxetine inhibited the stress axis in zebrafish. Intermediate concentrations of diazepam suppressed the stress response as measured by cortisol levels, whereas fluoxetine inhibited cortisol increase at concentrations similar to those found in the environment. These data suggest that the presence of psychoactive drugs in aquatic ecosystems could cause neuroendocrine dysfunction in fish.

Chemical communication of handling stress in fish

We investigated whether juveniles of the nocturnal fish jundiá (Rhamdia quelen) and the diurnal fish Nile tilapia (Oreochromis niloticus) are able to chemically communicate stress to conspecifics. Groups of 8 fish were reared in tanks under recirculated water (water exchanged among all the tanks) for each species. Fish were handled in half of the tanks (stressor fish) and whole-body cortisol concentrations were compared among handled fish, non-handled fish exposed to water from the handled fish, and non-handled control fish held with no water communication. For each treatment cortisol concentrations were determined before exposure to the stressor (basal levels) and after 1, 2, 4, 8, and 24h. Basal levels of cortisol confirmed fish were unstressed in the beginning of the experiment. Cortisol was increased in the stressor fish 1h after handling. Fish receiving water from the stressor fish increased cortisol levels later (2h after the stressor fish were handled). As the isolated control group maintained cortisol levels unchanged throughout the experiment, we concluded that some chemical factor was released by the stressed fish in the water and thus stressed the conspecifics. This pattern was similar for both unrelated species, thus suggesting that this communication might have evolved earlier in fish and reinforcing the biological value of this kind of information.

Alcohol impairs predation risk response and communication in zebrafish.

The effects of ethanol exposure on Danio rerio have been studied from the perspectives of developmental biology and behavior. However, little is known about the effects of ethanol on the prey-predator relationship and chemical communication of predation risk. Here, we showed that visual contact with a predator triggers stress axis activation in zebrafish. We also observed a typical stress response in zebrafish receiving water from these conspecifics, indicating that these fish chemically communicate predation risk. Our work is the first to demonstrate how alcohol effects this prey-predator interaction. We showed for the first time that alcohol exposure completely blocks stress axis activation in both fish seeing the predator and in fish that come in indirect contact with a predator by receiving water from these conspecifics. Together with other research results and with the translational relevance of this fish species, our data points to zebrafish as a promising animal model to study human alcoholism.

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.

Agrichemicals chronically inhibit the cortisol response to stress in fish.

We studied the stress response of Rhamdia quelen fingerlings at 45, 90, 135 and 180 d following acute exposure to agrichemicals. Herein, we report the novel observation that acute exposure of fingerling-aged fish to a methyl parathion-based insecticide (MPBI) and to a tebuconazole-based fungicide (TBF) induced chronic inhibition of the stress response. In contrast, fish exposed to an atrazine-simazine-based herbicide (ASBH) recovered the stress response on day 45, and fish exposed to a glyphosate-based herbicide (GBH) did not present stress response inhibition. Additionally, fish exposed to MPBI, GBH and ASBH showed lower survival rates and attained lower final weights. In the case of TBF, the presence of the stressful stimulus more strongly influenced the changes in the performance parameters than did the agrichemical exposure itself. An impairment of the cortisol response may seriously hamper the adaptive response and the ability to promote the necessary metabolic and ionic adjustments to respond to environmental stress.

Chemical communication of predation risk in zebrafish does not depend on cortisol increase

We investigated chemical cues among groups of zebrafish (Danio rerio) when communicating information about the risk of predation. We found that visual cues of the predator (tiger Oscar, Astronotus ocellatus) did not increase whole-body cortisol levels in groups of zebrafish but that water conditioned by these (donor) zebrafish stressed (target) conspecifics, thereby increasing whole-body cortisol. This finding was confirmed when these zebrafish groups were in different aquaria and communicated exclusively via water transfer. This result indicates that the stress induced in the target zebrafish does not depend on an increase in whole-body cortisol levels in the donor zebrafish. Because cortisol participation is rejected in this predation-risk communication, other chemicals from the stress systems should be investigated.

Bee Products Prevent Agrichemical-Induced Oxidative Damage in Fish

In southern South America and other parts of the world, aquaculture is an activity that complements agriculture. Small amounts of agrichemicals can reach aquaculture ponds, which results in numerous problems caused by oxidative stress in non-target organisms. Substances that can prevent or reverse agrichemical-induced oxidative damage may be used to combat these effects. This study includes four experiments. In each experiment, 96 mixed-sex, 6-month-old Rhamdia quelen (118±15 g) were distributed into eight experimental groups: a control group that was not exposed to contaminated water, three groups that were exposed to various concentrations of bee products, three groups that were exposed to various concentrations of bee products plus tebuconazole (TEB; Folicur 200 CE™) and a group that was exposed to 0.88 mg L−1 of TEB alone (corresponding to 16.6% of the 96-h LC50). We show that waterborne bee products, including royal jelly (RJ), honey (H), bee pollen (BP) and propolis (P), reversed the oxidative damage caused by exposure to TEB. These effects were likely caused by the high polyphenol contents of these bee-derived compounds. The most likely mechanism of action for the protective effects of bee products against tissue oxidation and the resultant damage is that the enzymatic activities of superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST) are increased.

Repeated stressors do not provoke habituation or accumulation of the stress response in the catfish Rhamdia quelen

Fish repeatedly experience stressful situations under experimental and aquaculture conditions, even in their natural habitat. Fish submitted to sequential stressors can exhibit accumulation or habituation on its cortisol response. We posed a central question about the cortisol response profiles after exposure to successive acute stressors of a similar and different nature in Rhamdia quelen. We have shown that successive acute stressors delivered with 12-h, 48-h, and 1-week intervals provoked similar cortisol responses in juvenile R. quelen, without any habituation or accumulation. The cumulative stress response is more associated to short acute stressors with very short intervals of minutes to hours. In our work, we used an interval as short as 12h, and no cumulative response was found. However, if the length of time between stressors is of a day or week as used in our work the most common and an expected phenomenon is the attenuation of the response. Thus, also, the absence of both accumulation of the stress response and the expected habituation is an intriguing result. Our results show that R. quelen does not show habituation or accumulation in its stress responses to repeated stressors, as reported for other fish species

Antioxidant activity of bee products added to water in tebuconazole-exposed fish

An experiment was conducted to evaluate the potential of honey, propolis, and bee pollen for the reversal of lipid peroxidation induced by tebuconazole (TEB) in South American catfish (Rhamdia quelen), in which the concentration of thiobarbituric acid reactive substances (TBARS), the activity of the antioxidant enzyme glutathione-S-transferase (GST) and the concentrations of non-enzymatic antioxidants, reduced glutathione (GSH), ascorbic acid, and non-protein thiols were assessed. Honey (0.125 g L-1) and bee pollen (0.05 g L-1) added to the water reverse the production of TBARS induced by TEB, while propolis demonstrated a pro-oxidant effect, inducing an increase in TBARS production. The data presented herein suggest that the addition of water to honey and bee pollen potentially protects against the oxidative stress caused by agrichemicals.