Acute sublethal exposure to ethiprole impairs physiological and oxidative status in the Neotropical fish Astyanax altiparanae.

Ethiprole, a phenylpyrazole insecticide, has been increasingly used in the Neotropical region to control stink bug pests in soybean and maize fields. However, such abrupt increases in use may have unintended effects on non-target organisms, including those inhabiting freshwater ecosystems. Here, we evaluated the effects of acute (96 h) sublethal exposure to ethiprole (up to 180 µg/L, which is equivalent to 0.013% of the recommended field dose) on biomarkers of stress in the gills, liver, and muscle of the Neotropical fish Astyanax altiparanae. We further recorded potential ethiprole-induced effects on the structural histology of A. altiparanae gills and liver. Our results showed that ethiprole exposure increased glucose and cortisol levels in a concentration-dependent manner. Ethiprole-exposed fish also exhibited higher levels of malondialdehyde and greater activity of antioxidant enzymes, such as glutathione-S-transferase and catalase, in both gills and liver. Furthermore, ethiprole exposure led to increased catalase activity and carbonylated protein levels in muscle. Morphometric and pathological analyses of the gills revealed that increasing ethiprole concentration resulted in hyperemia and loss of integrity of the secondary lamellae. Similarly, histopathological analysis of the liver demonstrated higher prevalence of necrosis and inflammatory infiltrates with increasing ethiprole concentration. Altogether, our findings demonstrated that sublethal exposure to ethiprole can trigger a stress response in non-target fish species, which may lead to potential ecological and economic imbalances in Neotropical freshwater systems.

Exposure to the herbicide atrazine induces oxidative imbalance, morphological damage and decreased survival in juvenile fish

Synthetic herbicides have been intensively used in weed control, although often involved in environmental contamination, critically affecting non-target species. However, never was investigated the effect of commercial formulation using atrazine on developing juvenile fish exposed for 35 days. Juveniles (Astyanax altiparanae) (n = 600) were assigned to the following ATZ-exposed groups: 0 (CTR-control), 0.56 (ATZ0.56), 1.00 (ATZ1.00), 1.66 (ATZ1.66) and 11.66 (ATZ11.66) µg/L. We found a 36.6% decrease in juvenile survival rate in the ATZ11.66 group compared to control and other groups. Juveniles from ATZ11.66 also showed hyperglycemia and increased cortisol levels. Increased the imbalance oxidative with an increase in malondialdehyde (MDA) and Carbonylated proteins levels markers in muscle, gills, and liver. We also found increased activity of the antioxidant enzymes superoxide dismutase (SOD) in gills and SOD and catalase (CAT) in muscles from ATZ11.66 fish, and increased glutathione S-transferase (GST) activities in the liver from all exposed groups compared to control. The morphological consequences of this were loss of secondary lamella integrity, increased mucus-secreting cells, hyperplasia, and lamellar fusion, as well as increased aneurysms percentage. The liver showed vascular congestion associated with endothelial hyperplasia, steatosis, and a decrease in the nuclei percentage. Our results showed that exposure to a commercial formulation of ATZ at 11.66 µg/L can be causing an imbalance in the oxidative markers and morphological damages and decreased survival in a juvenile Neotropical species of great ecological relevance and commercial interest.

Effects of subchronic exposure to environmentally relevant concentrations of the herbicide atrazine in the Neotropical fish Astyanax altiparanae.

Atrazine (ATZ) is among the most widely used herbicides in the world, and yet it has a potential to contaminate aquatic environments due to pesticide leaching from agricultural areas. In the Neotropical region, studies about the effects of this herbicide in native aquatic wildlife is scarce.Our study aimed at investigating the effects of a 30-day exposure to a commercial atrazine formulation on oxidative stress parameters, histopathology in testis and liver, and hormone levels in males and female of yellow-tailed tetra fish (Astyanax altiparanae). Adults were exposed to low but environmentally relevant concentrations of atrazine as follows: 0 (CTL-control), 0.5 (ATZ0.5), 1 (ATZ1), 2 (ATZ2) and 10 (ATZ10) µg/L. Our results showed decreased GST activity in gills in all groups of exposed animals and increased CAT activity in gills from the ATZ10 group. In the liver, there was an increase in lipid peroxidation in fish from ATZ1 and ATZ2 groups. Histological analysis of the liver showed increased percentage of sinusoid capillaries in ATZ2 fish, increased vascular congestion in ATZ1 and increased leukocyte infiltration in the ATZ10 group. Hepatocyte diameter analysis revealed a decrease in cell size in all groups exposed to ATZ, and a decrease in hepatocyte nucleus diameter in ATZ1, ATZ2 and ATZ10 groups. Endocrine parameters did not show significant changes following ATZ exposure, although an increase of triiodothyronine/thyroxine (T3/T4) ratio was observed in ATZ2 fish. Our results provide evidence that even low, environmentally relevant concentrations of ATZ produced oxidative damage and histological alterations in adult yellow-tailed tetra.

"Fearing the enemy": Growth and stress biomarker responses of sexually reversed Oreochromis niloticus in the presence of aquatic predatory insects.

Fishes can change their physiological responses when threatened by the presence of predators. Such physiological plasticity, however, usually implies costs that may impede organismal development and reproduction and reduce the ability to cope with other biotic and abiotic stresses. Here, we evaluated the growth and stress biomarker responses in sexually reversed Nile tilapia, Oreochromis niloticus, fingerlings indirectly threatened by the presence of the aquatic insect predator Belostoma anurum (Hemiptera: Belostomatidae). We also evaluated whether the presence of B. anurum would affect growth in fingerlings that received food containing the masculinizing hormone 17 α-methyltestosterone. The antioxidant responses were evaluated by measuring the activity of enzymes (e.g., superoxide dismutase, catalase, and glutathione-S-transferase). Oxidative stress biomarkers (e.g., malondialdehyde and nitric oxide) and blood glucose and lactate responses were also evaluated. Our results revealed that predator exposure did not affect growth in O. niloticus fingerlings reared in the presence or absence of the masculinizing hormone. However, sexually reversed tilapia fingerlings significantly increased not only the glucose and lactate blood levels, but also exhibited increased activities of superoxide dismutase and glutathione-S-transferases enzymes when threatened by the presence of B. anurum nymphs. Collectively, our findings indicate that despite not exhibiting reduced growth performance, sexually reversed tilapia fingerlings were physiologically stressed by the presence of the predator, which may reduce their ability to face environmental and abiotic stresses.

Curcuma longa as additive in the diet for Astyanax aff. bimaculatus.

The aim of the present study was to evaluate the effect of turmeric (Curcuma longa) as additive in the diet for Astyanax aff. bimaculatus. Fish (0.83 ± 0.04 g) were fed, for 60 days, with six diets containing 0.0, 20.0, 40.0, 60.0, 80.0, and 100.0 g turmeric kg-1 feed. There was an increasing linear effect of turmeric on the thickness of the muscular layer, and height and width of the folds of the intestine. In the liver, a quadratic effect was observed of turmeric on the percentage of hepatocyte cytoplasm and a decreasing linear effect on the percentage of sinusoid capillaries. A quadratic effect was also observed of turmeric on the liver glycogen. There was no effect of turmeric on the antioxidant activity in the liver, carcass composition or productive performance of the fish. Thus, we concluded that Curcuma longa has trophic effects on the epithelium and the muscular layer of the intestine of A. aff. bimaculatus. Additionally, low levels of Curcuma longa cause increased deposition of liver glycogen and high levels cause reduction.

Intestinal and liver morphometry of the Yellow Tail Tetra (Astyanax altiparanae) fed with oregano oil.

This study aimed to evaluate the effect of oregano oil on the intestinal and liver morphometry of yellow tail tetra, Astyanax altiparanae. Fish (1.46 ± 0.09 g) were kept in a 60-L aquaria, at a stocking density of 0.5 fi sh L-1. Six diets containing varying amounts of oregano oil were evaluated (0.0; 0.5; 1.0; 1.5; 2.0 and 2.5 g of oregano oil kg-1). At the end of 90 days, the fi sh were euthanised. Four intestines and four livers were collected per treatment, which were fi xed in Bouin and embedded in resin. For height and width folds, the absorption surface area and thickness of the muscular layer a positive linear effect of oregano oil was observed. A decrescent linear effect on the total number of goblet cells was also observed. For the cytoplasmic percentage of hepatocytes and liver glycogen, a positive linear effect of oregano oil was observed. There was a decreasing linear effect on the percentage of nuclei in the hepatocytes and capillaries. Thus, the oregano essential oil promotes increased absorption areas, modulates the amount of goblet cells involved in protecting the intestinal mucosa and promotes cytoplasmic increase with greater deposition of liver glycogen in yellow tail tetra.

Gelatin in replacement of bovine heart in feed training of Lophiosilurus alexandri in different water salinities.

The aim of this study was to evaluate commercial gelatin in the total replacement of bovine heart in feed training of "pacamã" Lophiosilurus alexandri in different water salinities. A completely randomized experimental design, in a 2 × 3 factorial arrangement, was performed using two types of moist ingredients (bovine heart and gelatin) and three water salinities (0.0; 2.0 and 4.0 g of salt L(-1)) with three replications. Juveniles (2.39 ± 0.08 cm standard length and 0.20 ± 0.03 g of weight) were conditioned to accept commercial diets by the technique of the gradual transition of ingredients. At the end of 36 days no differences were observed to weight gain, length gain and specific growth rate. The feed training efficiency was better (P < 0.05) with the gelatin use, 100.0%. There was a negative effect of salinity on the survival rate and management efficiency in the concentration of 4 g of salt L(-1), with values of 58.6 ± 12.0 % and 58.0 ± 12.0 %, respectively. Lophiosilurus alexandri juveniles could be feed-trained to accept commercial diets with gelatin in the total replacement of bovine heart in freshwater or salinity of 2 g of salt L(-1).

Transport of Astyanax altiparanae Garutti and Britski, 2000 in saline water / Transporte de Astyanax altiparanae Garutti and Britski, 2000 em água salinizada

Two experiments were performed. The first aimed to assess the tolerance of fingerlings Astyanax altiparanae to water salinity. Fish were exposed to salinity of 0, 3, 6, 9, 12 or 15 g NaCl L-1 for 96 hours. The fish mortality was 0%, in the levels of 0, 3 and 6 g L-1; 75% in the level of 9 g L-1and 100% at 12 and 15 g L-1 of common salt. The second experiment aimed to assess the parameters of water quality, mortality and blood glucose during transport. For this, A. altiparanae were stored in plastic bags at 22, 30 and 37 g of fish L-1 stocking densities and salinity of 0, 3, 6 and 9 g L-1, for. Fish showed similar mortality levels in the different salinities and stocking densities. The increase in fish density reduced the dissolved oxygen levels and salinity decreased the pH. The blood glucose levels were higher in those fish with 0 g L-1 salinity and higher stocking densities. The addition of salt to the water reduces the stress responses of A. altiparanae during transport.

Foram realizados dois experimentos. O primeiro teve o objetivo de avaliar a tolerância de alevinos de Astyanax altiparanae a salinidade da água. Os peixes foram submetidos às salinidades de 0, 3, 6, 9, 12 ou 15 g de NaCl L-1 durante 96 horas. A mortalidade dos peixes foi de 0%, nos níveis de 0, 3 e 6 g L-1; de 75% no nível de 9 g L-1 e de 100% em 12 e 15 g L- 1 de NaCl. No segundo experimento, objetivou-se avaliar os parâmetros de qualidade de água, mortalidade e a glicose sanguínea durante o transporte. Para isso, A. altiparanae foram estocados em sacos plásticos nas densidades de 22, 30 e 37 g de peixe L-1 e salinidades de 0, 3, 6 e 9 g de NaCl L-1. A mortalidade foi semelhante nas diferentes salinidades e densidades de estocagem. O aumento da densidade de peixes reduziu o nível de oxigênio dissolvido e a salinidade da água reduziu o pH. Os níveis de glicose sanguínea foram maiores nos peixes expostos a salinidade de 0 g L-1 e nas maiores densidades de estocagem. A adição de sal na água reduz as respostas de estresse em A. altiparanae durante o transporte.