Investigating the impact of Psidium guajava leaf hydroalcoholic extract in improving glutamatergic toxicity-induced oxidative stress in Danio rerio larvae.

Glutamate is one of the predominant excitatory neurotransmitters released from the central nervous system; however, at high concentrations, this substance may induce excitotoxicity. This phenomenon is involved in numerous neuropathologies. At present, clinically available pharmacotherapeutic agents to counteract glutamatergic excitotoxicity are not completely effective; therefore, research to develop novel compounds is necessary. In this study, the main objective was to determine the pharmacotherapeutic potential of the hydroalcoholic extract of Psidium guajava (PG) in a model of oxidative stress-induced by exposure to glutamate utilizing Danio rerio larvae (zebrafish) as a model. Data showed that treatment with glutamate produced a significant increase in oxidative stress, chromatin damage, apoptosis, and locomotor dysfunction. All these effects were attenuated by pre-treatment with the classical antioxidant N-acetylcysteine (NAC). Treatment with PG inhibited oxidative stress responsible for cellular damage induced by glutamate. However, exposure to PG failed to prevent glutamate-initiated locomotor damage. Our findings suggest that under conditions of oxidative stress, PG can be considered as a promising candidate for treatment of glutamatergic excitotoxicity and consequent neurodegenerative diseases.

Assessing the disparity: comparative toxicity of Copper in zebrafish larvae exposes alarming consequences of permissible concentrations in Brazil.

Copper (Cu) is a naturally occurring metal with essential micronutrient properties. However, this metal might also pose increased adverse environmental and health risks due to industrial and agricultural activities. In Brazil, the maximum allowable concentration of Cu in drinking water is 2 mg/L. Despite this standard, the impact of such concentrations on aquatic organisms remains unexplored. This study aimed to evaluate the toxicity of CuSO4 using larval zebrafish at environmentally relevant concentrations. Zebrafish (Danio rerio) larvae at 72 hr post-fertilization (hpf) were exposed to nominal CuSO4 concentrations ranging from 0.16 to 48 mg/L to determine the median lethal concentration (LC50), established at 8.4 mg/L. Subsequently, non-lethal concentrations of 0.16, 0.32, or 1.6 mg/L were selected for assessing CuSO4 -induced toxicity. Morphological parameters, including body length, yolk sac area, and swim bladder area, were adversely affected by CuSO4 exposure, particularly at 1.6 mg/L (3.31 mm ±0.1, 0.192 mm2 ±0.01, and 0.01 mm2 ±0.05, respectively). In contrast, the control group exhibited values of 3.62 mm ±0.09, 0.136 mm2 ±0.013, and 0.3 mm2 ±0.06, respectively. Behavioral assays demonstrated impairments in escape response and swimming capacity, accompanied by increased levels of reactive oxygen species (ROS) and lipid peroxidation. In addition, decreased levels of non-protein thiols and reduced cellular viability were noted. Data demonstrated that exposure to CuSO4 at similar concentrations as those permitted in Brazil for Cu adversely altered morphological, biochemical, and behavioral endpoints in zebrafish larvae. This study suggests that the permissible Cu concentrations in Brazil need to be reevaluated, given the potential enhanced adverse health risks of exposure to environmental metal contamination.

Permissible concentration of mancozeb in Brazilian drinking water elicits oxidative stress and bioenergetic impairments in embryonic zebrafish.

Mancozeb (MZ) is widely used as a fungicide in Brazil due to its effectiveness in combating fungal infections in plantations. However, its toxicity to non-target organisms, including aquatic organisms, has been reported in the literature. Recently, Brazilian legislation was updated to allow a concentration of 8 µg/L of MZ in drinking water (Ordinance GM/MS nº 888, of May 4, 2021). However, the safety of this concentration for aquatic organisms has not yet been put to the test. To address this gap, we conducted a study using zebrafish (Danio rerio) embryos at 4 hpf exposed to MZ at the concentration allowed by law, as well as slightly higher sublethal concentrations (24, 72, and 180 µg/L), alongside a control group. We evaluated various morphophysiological markers of toxicity, including survival, spontaneous movements, heart rate, hatching rate, body axis distortion, total body length, total yolk sac area, and total eye area. Additionally, we measured biochemical biomarkers such as reactive oxygen species (ROS) levels, lipid peroxidation, non-protein thiols (NPSH), and mitochondrial bioenergetic parameters. Our results showed that the concentration of 8 µg/L, currently permitted in drinking water according to Brazilian legislation, increased ROS production levels and caused alterations in mitochondrial physiology. Among the markers assessed, mitochondrial bioenergetic function appeared to be the most sensitive indicator of MZ embryotoxicity, as a decrease in complex I activity was observed at concentrations of 8 and 180 µg/L. Furthermore, concentrations higher than 8 µg/L impaired morphophysiological markers. Based on these findings, we can infer that the concentration of MZ allowed in drinking water by Brazilian environmental legislation is not safe for aquatic organisms. Our study provides evidence that this fungicide is a potent embryotoxic agent, highlighting the potential risks associated with its exposure.

Behavioral changes occur earlier than redox alterations in developing zebrafish exposed to Mancozeb.

As agriculture expands to provide food and wellbeing to the world's growing population, there is a simultaneous increasing concern about the use of agrochemicals, which can harm non-target organisms, mainly in the aquatic environment. The fungicide Mancozeb (MZ) has been used on a large-scale and is a potent inducer of oxidative stress. Therefore, there is an urgent need for the development of more sensitive biomarkers designed to earlier biomonitoring of this compound. Here we tested the hypothesis that behavioral changes induced by sublethal MZ concentrations would occur first as compared to biochemical oxidative stress markers. Embryos at 4 h post-fertilization (hpf) were exposed to Mancozeb at 5, 10 and 20 µg/L. Controls were kept in embryo water only. Behavioral and biochemical parameters were evaluated at 24, 28, 72, and 168 hpf after MZ exposure. The results showed that MZ significantly altered spontaneous movement, escape responses, swimming capacity, and exploratory behavior at all exposure times. However, changes in ROS steady-stead levels and the activity of antioxidant enzymes were observable only at 72 and 168 hpf. In conclusion, behavioral changes occurred earlier than biochemical alterations in zebrafish embryos exposed to MZ, highlighting the potential of behavioral biomarkers as sensitive tools for biomonitoring programs.