Acute exposure of embryo, larvae and adults of Danio rerio to fipronil commercial formulation reveals effects on development and motor control.

The insecticide fipronil, one of the main pesticides used in Brazil, is often detected in natural aquatic environments, and causes neuronal hyperexcitation by inhibiting GABAergic neurotransmission, leading to putative alterations in behaviour and development. This work sought to analyse the toxicity of formulated Regent® 800WG (80% fipronil) on development (fish embryo toxicity test, FET), morphology, and swimming behaviour of larvae and adults of zebrafish (Danio rerio). FET was performed following OECD236 guidelines at concentrations ranging from 0.002 to 1600 µg.L-1 of formulated Regent® 800WG. Adults were exposed to 0.2, 2 and 20 µg.L-1 of the product for 24 and 96 h, and were submitted to the light-dark, novel tank and swimming endurance tests No lethal parameters were observed in larvae, but in concentrations above 400 µg.L-1, there was shortening of the body axis and decreased swimming behavior. In adults, exposure to the pesticide did not lead to changes in free swimming parameters. However, a marked decrease of swimming endurance was observed at all experimental treatments, although probably not in consequence of energetic depletion, since baseline blood glucose levels and condition factor were similar at all conditions. Furthermore, zebrafish adults did not show their natural preference for the dark environment. The pesticide likely has anxiolytic effects on zebrafish, as well as a compromising effect on locomotor control, illustrating that behavioural changes, which could affect activities on the natural environment, such as escape and predation, may occur even in environmentally relevant concentrations of this pollutant.

Chromium accumulation and biomarker responses in the Neotropical fish Prochilodus lineatus caged in a river under the influence of tannery activities.

In order to assess the effects of tannery effluents (TE) in organism health, juveniles of Prochilodus lineatus were submitted to in situ tests at four different river locations: site A - upstream of the tannery; site B - next to the tannery; and sites C and D - downstream of the tannery. After 96 h exposure in the river, samples of fish tissue, river water and sediment were collected in order to quantify chromium (Cr) concentrations. Tissue samples were used to assess the activity of ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST), the content of glutathione (GSH) and metallothionein (MT) and the occurrence of lipid peroxidation (LPO) and DNA damage. Higher Cr concentrations were detected in the water and sediments from site B and in the liver of fish confined at site B, compared to the other sites. Fish caged at site B demonstrated higher levels of liver MT and hepatic EROD activity in relation to fish caged at the other sites. Moreover, fish from site B presented increased liver and branchial GST activities, as well as more GSH in the liver, than fish from site A. There were no significant variations in the occurrence of LPO and DNA damage among fish caged at the different sites. Thus, TE increased Cr levels in the water, sediments, and fish livers and stimulated the synthesis of MT and GSH and the activities of EROD and GST. In conclusion, TE affect the quality of the river and promote changes in biochemical biomarkers and Cr accumulation in P. lineatus.

Biomarkers of waterborne copper exposure in the Neotropical fish Prochilodus lineatus.

The main goal of the present study was to investigate the effects of acute exposure to copper (Cu) using a Neotropical freshwater fish as sentinel species through multi biomarkers analysis at different biological levels. Juveniles of Prochilodus lineatus were kept under control condition (no Cu addition in the water) or exposed to environmentally relevant concentrations of waterborne Cu (5, 9 and 20µgL(-1)) for 96h. These concentrations were selected to bracket the current Brazilian water quality criteria for Cu in fresh water (9 and 13µgL(-1) dissolved copper). Endpoints analyzed included ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, reduced glutathione (GSH) and metallothionein-like protein (MT) concentration, lipid peroxidation (LPO) level, tissue damage index, and incidence of free melano-macrophages (FMM) and melano-macrophage centers (MMC) in the liver. They also included DNA damage (frequency of nucleoids per comet class, number of damaged nucleoids per fish and DNA damage score) in erythrocytes, as well as muscle and brain acetylcholinesterase (AChE) activity and behavioral parameters (swimming distance and velocity, time spent swimming and swimming activity in the upper and lower layers of the water column). Fish exposed to any of the Cu concentrations tested showed increased liver MT concentration and LPO level, higher number of damaged nucleoids in erythrocytes per fish, and inhibited muscle AChE activity. Also, increased liver SOD activity was observed in fish exposed to 9 and 20µgL(-1) Cu. Fish exposed to 5 and 9µgL(-1) Cu spent lower amount of time swimming. Fish exposed to 9µgL(-1) Cu showed increased swimming distance and velocity while those exposed to 20µgL(-1) Cu had lower swimming distance and velocity, as well as, spent less time swimming in the lower layer of the water column when compared to those kept under control condition. These findings indicate that Cu exposure at environmentally relevant concentrations (below or close to the current Brazilian water quality criteria) induced significant biological (histological, biochemical and genetic) and ecological (swimming and exploratory abilities) damages in the Neotropical fish P. lineatus. They also suggest that MT concentration, DNA damage (comet assay), LPO (TBARS method), SOD and AChE activity, together with swimming behavior analyses are potential biomarkers to assess and monitor areas impacted by Cu in fresh water.

Histological changes in gills of two fish species as indicators of water quality in Jansen Lagoon (São Luís, Maranhão State, Brazil).

Water quality of the Jansen Lagoon (São Luís, Maranhão State, Brazil) was assessed through histological biomarkers and microbiological parameters. To this end, 29 fish specimens (11 Centropomus undecimalis and 18 Sardinella sp) and eight water samples were collected during the rainy and dry periods of 2013. The lagoon water showed thermotolerant coliform indices above the limit set forth in CONAMA Resolution 357/2005. Histological changes observed in the gills were: lifting of the respiratory epithelium, hyperplasia of the lamellar epithelium, incomplete and complete fusion of several lamellae, disorganization of the lamellae, congestion of blood vessels, aneurysms, hypertrophy of the respiratory epithelium, hemorrhage and rupture of the lamellar epithelium and parasite. The histological alteration index (HAI) average value to Sardinella sp was 31.8 and to C. undecimalis was 22.2. The average HAI value in both species corresponds to category 21-50, with tissue injuries being classified from moderate to severe. The presence of histological injuries and the HAI values indicate that the fish sampled from the Jansen Lagoon are reacting to non-specific xenobiotics present at the site.

Acute effects of sediments taken from an urban stream on physiological and biochemical parameters of the neotropical fish Prochilodus lineatus.

Juveniles of Prochilodus lineatus were exposed to sediments collected from one of five sites along an urban stream into which various types of contaminants are discharged. After 24 or 96 h fish were examined and the results compared with those from control groups (fish exposed only to water, for the same period). Plasma ion levels varied significantly and fish exposed to site 5 sediment showed a transient increase in both sodium and chloride concentrations. Plasma glucose was significantly higher in fish exposed to sediment from sites 2 and 5. The higher liver glutathione-S-transferase activity registered in fish exposed to sediment from sites 1, 4 and 5 suggests the presence of organic contaminants at these sites and the enhancement of liver catalase activity in fish exposed to sediment from sites 3 and 4 may be due to contaminant-mediated oxyradical production. The overall results revealed that sites 4 and 5 are more severely contaminated, probably due to organic contaminants from agricultural sources and municipal landfill.