Effects of climate change and mixtures of pesticides on the Amazonian fish Colossoma macropomum.

Several studies highlighted the complexity of mixing pesticides present in Amazonian aquatic environments today. There is evidence that indicates that ongoing climate change can alter the pattern of pesticide use, increasing the concentration and frequency of pesticide applications. It is known that the combination of thermal and chemical stress can induce interactive effects in aquatic biota, which accentuates cell and molecular damage. However, considering that the effects of climate change go beyond the increase in temperature the objective of this study was to evaluate the effect of climate change scenarios proposed by 6 th IPCC report and a mixture of pesticides on the tambaqui (Colossoma macropomum). The hypothesis of this study is that the negative effects will be accentuated by the combination of an extreme climate changes scenario and a mixture of pesticides. To test the hypothesis, juvenile tambaqui were exposed to a combination of four pesticides (chlorpyrifos, malathion, carbendazim and atrazine) in two scenarios, one that simulates current environmental conditions and another that predicted the environmental scenario for the year 2100. Fish were subjected to the experimental conditions for 96 h. At the end of the experiment, samples of blood, gills, liver, brain, and muscle were obtained for hematological, genotoxic, biochemical, and histopathological analyses. The results demonstrate that environmentally realistic concentrations of pesticides, when mixed, can alter the biochemical responses of tambaqui. The extreme scenario promotes hematological adjustments, but impairs branchial antioxidant enzymes. There is an interaction between the mixture of pesticides and the extreme scenario, accentuating liver tissue damage, which demonstrates that even increased activity of antioxidant and biotransformation enzymes were not sufficient to prevent liver damage.

Malathion alters the transcription of target genes of the tumour suppressor tp53 and cancerous processes in Colossoma macropomum: Mechanisms of DNA damage response, oxidative stress and apoptosis.

Different classes of pesticides such as fungicides, herbicides, and insecticides, can induce differential expression of genes that are involved in tumorigenesis events in fish, including the expression of tumor suppressor tp53. The degree and duration of the stressful condition is decisive in defining which tp53-dependent pathway will be activated. Herein we evaluate the target genes expression that participates in the regulation pathway of the tumor suppressor tp53 and in the cancerous processes in tambaqui after exposure to malathion. Our hypothesis is that malathion promotes a gene response that is differentially regulated over time, with positive regulation of tp53 target genes related to the apoptotic pathway and a negative regulation of genes that promote antioxidant responses. The fish were exposed to a sublethal concentration of the insecticide for 6 and 48 h. Liver samples were used to analyze the expression of 11 genes using real-time PCR. Overall, the malathion promoted over time increases in tp53 expression and differential expression of tp53 related genes. The exposure resulted in the activation of damage response related genes, caused a positive expression of atm/atr genes. The pro-apoptotic gene bax was up-regulated and the anti-apoptotic bcl2 was down-regulated. Increased expression of mdm2 and sesn1 in the first hours of exposure and no effect on the antioxidant genes sod2 and gpx1 were also observed. We also witnessed an increase in the expression of the hif-1α gene, with no effect on ras proto-oncogene. The extension of this stressful condition accentuated tp53 transcription, and minimized the levels of mdm2, sens1 and bax; however, it down regulated the levels of bcl2 and the bcl2/bax ratio, which indicates the maintenance of the apoptotic response to the detriment of an antioxidant response.

Digestive tube of an herbivorous fish (Rhytiodus microlepis) from the Amazonian floodplain lakes: A morphological and histochemical study.

This study describes the morphology and histochemistry of the digestive tube of the herbivorous fish Rhytiodus microlepis, which is commonly found in the Amazonian floodplain lakes, Brazil. Thirty-eight fish were used in this study. The digestive tube was measured and dissected for anatomical description, and stained with (i) haematoxylin and eosin, for histological analyses; (ii) periodic acid-Schiff, for detected neutral mucins; and (iii) Alcian Blue (AB) pH 1.0 and 2.5 for acid and sulphated acid mucins. The oesophagus, with its secretory cells and taste buds, is adapted for lubrication of vegetable matter, resistance to friction and reception of stimuli. The stomach is U-shaped and rich in gastric glands, particularly in cardiac and fundic regions, which are adapted to receive large amounts of food, and promotes the digestion. The intestine comprises more than 70% of the digestive tube. All portions of the intestine, except the rectum, have the same pattern of mucosal folds, and the goblet cells reacted positively to all histochemical methods (PAS, AB 1.0 and 2.5), while rodlet cells reacted only to periodic acid-Schiff (PAS) staining. Neutral mucins and sulphated acid mucins predominated in the anterior portion of the intestine and a high concentration of carboxylated acid mucosubstances were present in the other portions. The rectum showed a strong reaction to all histochemical methods. The muscular layer of the intestine consists of three layers, which showed features presumably related to the rigid nature of the food. A simple squamous epithelium constitutes the serous layer over the entire length of the tube.

Distribution of the acanthocephalan Neoechinorhynchus buttnerae and semiquantitative analysis of histopathological damage in the intestine of tambaqui (Colossoma macropomum).

In this paper, we have described for the first time a semiquantitative method to evaluate histopathological damage, taking the degree of Neoechinorhynchus buttnerae attachment to the intestinal wall of the tambaqui (Colossoma macropomum), an important species in Brazilian aquaculture, into account. Twelve specimens of tambaqui were collected from a fish farm. Their bowels were removed and divided into seven morphologically distinct portions according to density and distribution of the parasite studies. Fragments from each fraction were histologically processed and analyzed. There was a clear preference on the part of N. buttnerae for the intermediate regions of the intestinal tube, where the highest densities were recorded. The intensity of damage to the host, estimated by calculating the Histopathological Alteration Index (HAI), showed severe and irreversible changes only where the parasite had its proboscis penetrated into the intestine wall.

Potential risks of natural mercury levels to wild predator fish in an Amazon reservoir.

Mercury (Hg) is a toxic metal that bioaccumulates in aquatic organisms and along food chain. Many studies have reported the problem of mercury exposure in aquatic systems from Amazon basin, but very few have focused on the potential risks to wild fish. The present study reports the bioaccumulation of mercury and alterations in target organs of the predator fish Hoplias malabaricus (traíra) from Samuel reservoir, Amazon basin, Northern Brazil. About 18% of fish had mercury levels in muscle exceeding the safe limit for ingestion through food, established by WHO (0.5 µg Hg g(-1)). Fish were separated in two groups according to mercury bioaccumulation in liver (<0.2 µg Hg g(-1)-group I and >0.2 µg Hg g(-1)-group II) for biomarker comparisons. Catalase activity and number of macrophage centers were statistically higher in group II, confirming the potential of Hg to interfere with redox balance and to recruit defense cells to the liver. Conversely, erythrocyte nuclear alterations were less frequent in group II, indicating a more rigorous selection of erythrocytes or hormesis pattern of response. Glutathione S-transferase activity, lipid peroxidation, and histopathological analyses were not statistically different in the liver and gills of both groups. Comparison of lipid peroxidation levels of these fish with others captured in Southern Brazil during another study and the high incidence of morphological alterations in the liver and gills suggest that the bioaccumulation of mercury during continuous exposure is posing potential risks to the species.