Bioconcentrations of herbicides used in sugarcane crops in tilapia (Oreochromis niloticus) and the risk for human consumption.

The practice of intensive herbicide use in the sugarcane industry has a high risk of compromising the quality of the water and the organisms that live there due to losses through runoff, leaching and other processes. In this work, the dynamics of four herbicides present in three different mixtures were evaluated through their incorporation and elimination in the muscle tissue of tilapia (Oreochromis niloticus). The highest mean values of bioconcentration factors were 1.730 for ametryn, 0.891 for tebuthiuron, 0.322 for hexazinone and 4.783 for diuron. Diuron presented the highest risk regarding the consumption of tilapia fillets by the population. However, considering that the fish would reach maximum levels of diuron when exposed to extremely high concentrations, an individual weighing 70 kg would need to ingest approximately 1.5 kg of this food product to surpass the acceptable daily intake of 0.007 mg kg-1 body weight. It was concluded that the risk of injury to the population consuming tilapia fillets from fish exposed to herbicides in water arising from sugarcane activities is very low. According to the risk estimation performed in this work, which is substantiated by the assumptions of the World Health Organization and the International Life Sciences Institute, there is a low risk of injury to the population consuming tilapia fillets from fish exposed to water containing herbicides in concentrations arising from sugarcane activities. However, as the risk was estimated from laboratory conditions, caution should be taken where herbicide applications are carried out with high frequency near water bodies, as the consumption of fish from these areas is quite common.

Multiresidue Method for Quantification of Sulfonamides and Trimethoprim in Tilapia Fillet by Liquid Chromatography Coupled to Quadrupole Time-of-Flight Mass Spectrometry Using QuEChERS for Sample Preparation.

A multiresidue method for detecting and quantifying sulfonamides (sulfapyridine, sulfamerazine, sulfathiazole, sulfamethazine, sulfadimethoxine, sulfamethoxazole, and sulfamethoxypyridazine) and trimethoprim in tilapia fillet (Oreochromis niloticus) using liquid chromatography coupled to mass spectrometry was developed and validated. The sample preparation was optimized using the QuEChERS approach. The chromatographic separation was performed using a C18 column and 0.1% formic acid in water and acetonitrile as the mobile phase in the isocratic elution mode. Method validation was performed based on the Commission Decision 2002/657/EC and Brazilian guideline. The validation parameters evaluated were linearity (r ≥ 0.99); limits of detection (LOD) and quantification (LOQ), 1 ng·g-1 and 5 ng·g-1, respectively; intraday and interdays precision (CV lower than 19.4%). The decision limit (CCα 102.6-120.0 ng·g-1 and 70 ng·g-1 for sulfonamides and trimethoprim, respectively) and detection capability (CCß 111.7-140.1 ng·g-1 and 89.9 ng·g-1 for sulfonamides and trimethoprim, respectively) were determined. Analyses of tilapia fillet samples from fish exposed to sulfamethazine through feed (incurred samples) were conducted in order to evaluate the method. This new method was demonstrated to be fast, sensitive, and suitable for monitoring sulfonamides and trimethoprim in tilapia fillet in health surveillance programs, as well as to be used in pharmacokinetics and residue depletion studies.

Depletion study, withdrawal period calculation and bioaccumulation of sulfamethazine in tilapia (Oreochromis niloticus) treated with medicated feed.

The residue depletion of sulfamethazine (SMZ) was evaluated in tilapia (Oreochromis niloticus) after 11 days of administration of medicated feed containing SMZ, at the dose of 422 mg/kg body weight (bw). The determination of SMZ in feed and tilapia fillet was carried out using the QuEChERS approach for sample preparation, and high performance liquid chromatography with diode array detector (HPLC-DAD) and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS) for quantitation, respectively. Both methods were validated based on international and Brazilian guidelines and shown to be suitable for the intended purposes. The withdrawal period to reach the maximum residue level (MRL) of 100 µg/kg, according to the European Union (EU) legislative framework to all substances belonging to the sulfonamide (SA) group (EU, 2010), was 10 days (260 °C-day). After treatment, the maximum level of SMZ accumulation in the tilapia muscle was 1.6 mg/kg. SMZ was shown to be quickly excreted by tilapia. Thus, considering the acceptable daily intake of SMZ established by the Codex Commission (0-0.05 mg/kg bw), and a factor of 5 times the upper amount of fish consumption in Brazil (38 kg/year), this study showed that there is a low risk of adverse effects to consumers. This study offers subsidies not only for the establishment of public policies with regard to the use of veterinary drugs currently not allowed in a country by their legal legislative framework for fish farming, but also to fish producers for the proper handling to ensure safe fish fillets.

Mixtures of diflubenzuron and p-chloroaniline changes the activities of enzymes biomarkers on tilapia fish (Oreochromis niloticus) in the presence and absence of soil.

The insecticide Diflubenzuron (DFB), used by many fish farming, when metabolized or degraded produces the extremely toxic compound p-chloroaniline (PCA). Once in the aquatic environment, these compounds can form mixtures and their bioavailability depends on factors such as the presence of soil. The toxic effects of the isolated compounds and their mixtures in the proportions: 75%, 50%, and 25% of PCA were analyzed in tilapia (Oreochromis niloticus) in the presence and absence of soil after 96h. The enzymes catalase (CAT), acid (AcP) and alkaline (AlP) phosphatases and alanine (ALT) and aspartate (AST) aminotransferases of the liver of the tilapia (Oreochromis niloticus) were used as biomarkers. DFB and the mixture containing 75% of this compound did not present high toxicity to fish; however, 25mg/L of PCA alone and 15mg/L of the mixture with 75% of this compound promoted 50% mortality of tilapia (Oreochromis niloticus). In the presence of soil, these toxicity values decreased to 37 and 25mg/L, respectively. Independent of the presence of soil, a synergistic effect was observed when the proportion of PCA was 75% and to the mixture, with 25% PCA was observed the antagonistic effect. Different concentrations of the compounds and their mixtures induced CAT activity independently of the presence of soil. Additionally, increases in phosphatases and transaminases activities were observed. In some cases, the enzymes also had their activities decreased and the dose-dependence effects were not observed. This research showed that the presence of soil influenced the toxicity of the compounds but not altered interaction type among them. Diflubenzuron, p-chloroaniline, and mixtures thereof caused disorders in enzymes important for the health of tilapia (Oreochromis niloticus).

The sugarcane herbicide ametryn induces oxidative stress and developmental abnormalities in zebrafish embryos.

Ametryn (AMT) is one of the most widely used herbicides in tropical sugarcane crops, the main culture of São Paulo State, Brazil. It is known as a diffuse pollutant, being found in surface water and sediment of water bodies adjacent to the crop fields. In the present study, the toxicity of AMT to zebrafish (Danio rerio) embryos was evaluated using developmental and biochemical endpoints. At the biochemical level, lactate dehydrogenase responded at the lowest concentration tested (4 µg L-1) indicating a high demand of energy required to cope with the stress condition. Antioxidant enzyme levels were changed at intermediate/high concentrations while oxidative damage (lipid peroxidation) was observed at the last concentration tested (10 mg L-1). This suggests that, like for other herbicides from the triazine group, oxidative stress is a major pathway of toxicity for AMT. Several developmental effects such as oedemas and tail deformities were also observed. The 96 h-EC50 values calculated for different developmental parameters were between 17 and 29 mg L-1. AMT also affected hatching (96 h-EC50 = 22.5 mg L-1) and positioning in the water column (96 h-EC50 = 13.2 mg L-1). In a previous work of the group, lethal toxicity of AMT showed to be much higher to adults than to embryos. However, in the present work, sublethal endpoints assessed suggest that important effects are observed at lower concentrations, improving the sensitivity of the embryo test. Moreover, in this work, sublethal effects were observed for concentrations in the same range as the ones found in the environment, and thus, given that this chemical is widely used in tropical fields, a refined evaluation of risk should be performed based on the monitoring of sublethal and long-term parameters and considering mixture scenarios.

Chitosan nanoparticles loaded the herbicide paraquat: the influence of the aquatic humic substances on the colloidal stability and toxicity.

Polymeric nanoparticles have been developed for several applications, among them as carrier system of pesticides. However, few studies have investigated the fate of these materials in the environment in relation to colloidal stability and toxicity. In nature, humic substances are the main agents responsible for complexation with metals and organic compounds, as well as responsible for the dynamics of these nanoparticles in aquatic and terrestrial environments. In this context, the evaluation of the influence of aquatic humic substances (AHS) on the colloidal stability and toxicity of polymeric nanoparticles of chitosan/tripolyphosphate with or without paraquat was performed. In this study, the nanoparticles were prepared by the ionic gelation method and characterized by size distribution measurements (DLS and NTA), zeta potential, infrared and fluorescence spectroscopy. Allium cepa genotoxicity studies and ecotoxicity assays with the alga Pseudokirchneriella subcapitata were used to investigate the effect of aquatic humic substances (AHS) on the toxicity of this delivery system. No changes were observed in the physical-chemical stability of the nanoparticles due to the presence of AHS using DLS and NTA techniques. However some evidence of interaction between the nanoparticles and AHS was observed by infrared and fluorescence spectroscopies. The ecotoxicity and genotoxicity assays showed that humic substances can decrease the toxic effects of nanoparticles containing paraquat. These results are interesting because they are important for understanding the interaction of these nanostructured carrier systems with species present in aquatic ecosystems such as humic substances, and in this way, opening new perspectives for studies on the dynamics of these carrier systems in the ecosystem.

Acute toxicity and bioconcentration of fungicide tebuconazole in zebrafish (Danio rerio).

This research work investigated the bioconcentration of tebuconazole [(±)-α-[2-(4-chlorophenyl)ethyl]-α-(1,1-dimethylethyl)-1H-1,2,4-triazole-1-ethanol] fungicide in zebrafish (Danio rerio) under laboratory conditions and a first-order kinetic pesticide dissipation in the water. The concentrations of tebuconazole fitted to an equivalent nonlinear kinetic type model which allowed the calculation of the following parameters: bioconcentration factor (38.80 L kg(-1) ), time to reach maximum fish concentration (6 days), maximum concentration in fish (0.0075 µg mg(-1) ), half-life in fish (24 days) and time needed for the fish to eliminate 95% of the maximum concentration (105 days). These calculations permitted the establishment of theoretical reference limit values for human consumption of fish and the establishment of safe limits for the water pesticide concentration. The data would also be useful in safe strategies associated with fishery activities that are conducted in aquatic regions close to crops using tebuconazole. The information will contribute to enlarge the tebuconazole toxicokinetics database of aquatic organisms.