Non-lethal biomarkers as promising tools for fish health assessment: In situ exposure to bifenthrin as a case study.

Non-lethal biomonitoring should provide an innovative approach to establish bioethical protocols for the management of both aquaculture and wild fisheries resources. We aimed to assess non-lethal biomarkers in Piaractus mesopotamicus caged in a rice field during a bifenthrin (BF) application. We analyzed parameters related to the immune system, energy metabolism and oxidative stress in fish skin mucus and blood plasma. Fish exposed to BF showed a significant increase in skin mucus glucose levels and the enzymatic activities of protease, alkaline phosphatase and superoxide dismutase. Regarding plasmatic parameters, BF increased the levels of glucose, total protein and albumin, but decreased triglycerides. In addition, increased activities of lysozyme and alkaline phosphatase were found in the blood plasma of exposed fish. Our results indicated an increased energy demand, altered immune function and a mild oxidative stress response in fish exposed in situ to BF. We have shown that skin mucus and blood plasma are very promising matrices for the development of non-lethal biomarkers to assess fish health in a stressed environment.

Nanosilica and copper ecotoxicity in Gambusia holbrooki fish.

When silica nanoparticles (SiNP) reach the water bodies interact with the already existing pollutants in the environments. This study aimed to evaluate the ecotoxicity of SiNP under the presence/absence of Cu in mosquitofish (Gambusia holbrooki). Fish were exposed to 0, 10 and 100 mg SiNP L-1, alone or mixed with Cu (0.25 mg L-1). After 96 h, the amount of colony forming units (CFU) of bacteria living on the skin mucus was analysed, and oxidative stress, tissue damage enzymes, and neurotoxicity were evaluated. We observed a reduction in CFU when Cu was present in the media. The liver was the target organ, evidencing a decrease in tissue damage enzymatic activities, activation of the antioxidant system in all treatments, and lipid oxidative damage when the SiNP and Cu were mixed. Overall, SiNP ecotoxicity was proved, which could also be enhanced by the presence of ubiquitous elements such as metals.

Metallic, metal oxide, and metalloid nanoparticles toxic effects on freshwater microcrustaceans: An update and basis for the use of new test species.

In this article, we performed a literature review on the metallic, metal oxide, and metalloid nanoparticles (NP) effects on freshwater microcrustaceans, specifically focusing on (i) the main factors influencing the NP toxicity and (ii) their main ecotoxicological effects. Also, given that most studies are currently developed on the standard test species Daphnia magna Straus, we analyzed (iii) the potential differences in the biological responses between D. magna and other freshwater microcrustacean, and (iv) the ecological implications of considering only D. magna as surrogate of other microcrustaceans. We found that NP effects on microcrustaceans depended on their intrinsic properties as well as the exposure conditions. Among the general responses to different NP, we identified body burial, feeding inhibition, biochemical effects, metabolic changes, and reproductive and behavioral alterations. The differences in the biological responses between D. magna and other freshwater microcrustacean rely on the morphology (size and shape), ecological traits (feeding mechanisms, life cycles), and intrinsic sensitivities. Thus, we strongly recommend the use of microcrustaceans species with different morphological, physiological, and ecological characteristics in future ecotoxicity tests with NP to provide relevant information with regulation purposes regarding the discharge of NP into aquatic environments. PRACTITIONER POINTS: Nanoparticles effects depend on intrinsic and external factors. Nanoparticles affect the morphology, physiology, and behavior. Effects on Daphnia differ from other microcrustaceans. The use of more diverse test species is suggested.

Low temperature stress in a cultured fish (Piaractus mesopotamicus) fed with Pyropia columbina red seaweed-supplemented diet.

This study aimed to analyze the cold stress effects (in terms of hematology, energy reserves, and oxidative stress) in Piaractus mesopotamicus (pacú) and their mitigation by a Pyropia columbina red seaweed-supplemented diet. For this purpose, juvenile fish were fed with a control (CD) or a red seaweed-supplemented diet (RD) for 60 days, and then, the animals were exposed to a low temperature (14 °C) and a control temperature (24 °C) for 24 h. The cold shock generated an increase of hemoglobin levels in fish fed with both diets. In CD-fed fish, plasmatic triglycerides, cholesterol, and hepatic glycogen decreased after the thermal shock; meanwhile, the animals fed with RD showed decreased hepatic proteins, but increased cholesterol and hepatic glycogen. Regarding oxidative stress, antioxidant enzymes augmented their activity in the liver, intestine, and gills; meanwhile, lipid oxidative damage was observed in the liver and intestine of fish exposed to 14 °C and fed with both diets. Pacú was sensitive to cold shock, but no mitigation effects were observed in fish fed with the supplemented diet. Further research should target higher concentrations of P. columbina in supplemented diets to take advantage of this valuable resource.

Nanoparticles Toxicity in Fish Models.

The increasing production and use of nanoparticles (NP) have raised concerns regarding the potential toxicity to human and environmental health. In this review, we address the up to date information on nanotoxicity using fish as models. Firstly, we carried out a systematic literature search (articles published up to February 2019 in the Scopus database) in order to quantitatively assess the scientific research on nanoparticles, nanotoxicity and fish. Next, we carried out a narrative synthesis on the main factors and mechanisms involved in NP toxicity in fish. According to the bibliometric analysis, there is a low contribution of scientific research on nanotoxicity compared with the general nanoparticles scientific production. The literature search also showed that silver and titanium NP are the most studied nanomaterials and Danio rerio is the fish species most used. In comparison with freshwater fish, the effects of nanomaterials on marine fish have been little studied. After a non-systematic literature analysis, we identified several factors involved in nanotoxicity, as well as the effects and main toxicity mechanisms of NP on fish. Finally, we highlighted the knowledge gaps and the need for future research.

Toxicological Effects Induced by Silver Nanoparticles in Zebra Fish (Danio Rerio) and in the Bacteria Communities Living at Their Surface.

The antimicrobial activity of silver nanoparticles (AgNP) makes them useful in a wide range of products although their environmental impact is still uncertain. The main goal of this study was to evaluate short-term effects induced by AgNP on gills oxidative status and bacterial communities living at the skin mucus of zebrafish. Both the number of bacteria colony forming units and bacteria growth obtained from skin mucus were lower in all concentrations tested (25, 50 and 100 µg nAg/L). Besides, AgNP exposure caused a significant decrease in bacteria growth in zebrafish exposed to 100 µg nAg/L. AgNP accumulated in zebrafish gills at both highest concentrations tested, but this accumulation did not appear to result in oxidative stress. Overall the results indicated toxicological effects of AgNP on bacteria communities living at the zebrafish mucus surface. Although silver accumulation was verified in gills, no evidence of toxicity in terms of oxidative stress was found.

Evaluación de genotoxicidad a través de la frecuencia de Micronúcleos en eritrocitos de Piaractus mesopotamicus (pacúes) expuestos in vivo a nanopartículas de plata / Assessment of genotoxicity by Micronucleus frequency in Piaractus mesopotamicus erythrocytes exposed to silver nanoparticles in vivo

El uso creciente de nanomateriales en productos industriales y de consumo ha incrementado la preocupación mundial respecto a sus posibles efectos adversos en los sistemas biológicos. Como consecuencia de la falta de un marco legislativo y la ausencia de un consenso sobre los protocolos experimentales, las investigaciones ecotoxicológicas se llevan a cabo a un ritmo mucho más lento que la producción de nuevas nanopartículas. Por esta razón, existe una necesidad creciente de realizar estudios que aporten conocimiento sobre el riesgo de estos contaminantes emergentes de propiedades únicas. El objetivo del presente trabajo fue evaluar la frecuencia de micronúcleos (FMN) en eritrocitos de ejemplares juveniles de pacú (Piaractus mesopotamicus) expuestos a nanopartículas de plata (Nano-Ag) a las concentraciones de 0 μg·L-1 (control); 2,5 μg·L-1; 10 μg·L-1; y 25 μg·L-1, durante 24 horas. Se observó que la FMN se incrementó significativamente (p<0,01) en la concentración de 25 μg·L-1, mientras que no hubo diferencias significativas entre los grupos expuestos a 2,5 y 10 μg·L-1 y el control. Estos resultados sugieren que los eventos aneugénicos o clastogénicos podrían representar un posible mecanismo de toxicidad de Nano-Ag en esta especie.

The growing use of nanomaterials in consumer and industrial products has aroused global concern about possible adverse effects on biological systems. Due to the lack of a regulation framework and the absence of a consensus on the experimental protocols, ecotoxicological investigations are carried out much slower than the production of new nanoparticles. For this reason, there is a growing need for studies that provide knowledge about the risk of these emerging contaminants of unique properties. The objective of the present study was to evaluate the frequency of micronuclei (FMN) in erythrocytes of juvenile Piaractus mesopotamicus exposed to silver nanoparticles (nano-Ag; Nanotek SA) at concentrations of 0 μg·L-1 (control); 2.5 μg·L-1; 10 μg·L-1; and 25 μg·L-1, for 24 hours (n = 10 per treatment). The FMN show a significant increase (p <0.01) in fish exposed to 25 μg·L-1 of Nano-Ag, while there were no significant differences among the groups exposed to 2.5 and 10 μg·L-1 with the control. These results suggest that the aneugenics or clastogenics events may represent a possible mechanism of toxicity of Nano-Ag in this specie.

Histopathological changes in the gills and liver of Prochilodus lineatus from the Salado River basin (Santa Fe, Argentina).

This study evaluated the histopathological changes in gills and liver of Prochilodus lineatus inhabiting the Salado River basin. Fish were collected in four different sampling stations. The histological lesions in the tissues were examined under light microscopy and evaluated with quantitative analyses. The morphometric analysis of the gills showed a significant shortening of secondary lamellae and a lower percentage of area for gas exchange in fish from station 1 (an urban area, located near the mouth of the Salado River) in comparison with fish gills from the reference site (station 4, a relatively pristine area). Moreover, a significantly higher area occupied with necrotic foci and the occurrence of an important inflammatory response were observed in fish liver of station 1 than the samples caught from other stations. Thus, histopathological evidences showed differences among sites, which could be related to different environmental conditions.

Biochemical stress responses in tissues of the cichlid fish Cichlasoma dimerus exposed to a commercial formulation of endosulfan.

Median lethal concentration (LC(50)) and sublethal effects of the commercial endosulfan formulation Zebra Ciagro(®) on the fish Cichlasoma dimerus were studied. The 96-h LC(50) was estimated as 17.7 µg/L. In order to investigate sublethal effects, fish were exposed to 25% and 50% LC(1) (3.4 and 6.8 µg/L, respectively). Endosulfan (ED) significantly increased the hemoglobin concentration and white blood cell count after 96 h. Differential leukocytes count was also altered, due to an increase in the percentage of neutrophils in exposed fish. The hepatopancreatic tissue of fish under ED treatment showed a decrease in aspartate aminotransferase and alanine aminotransferase and an increase in alkaline phosphatase. Lipid peroxidation levels in the 6.8-µg/L ED-containing group were higher than those in control fish for all organs tested (gills, hepatopancreas, and brain).

Multiple biomarkers responses in Prochilodus lineatus allowed assessing changes in the water quality of Salado River basin (Santa Fe, Argentina).

This field study assessed water quality of Salado River basin by using a set of biomarkers in the fish Prochilodus lineatus. Multiple biomarkers were measured, including morphological indexes (condition factor, liver somatic index), hematological (red and white blood cells) and biochemical (glucose, total protein and cholinesterase activity) parameters. Besides, detoxication and oxidative stress markers (antioxidant enzymes, lipid peroxidation) were measured in liver, gills and kidney. Despite water quality assessment did not show marked differences among sites, biomarkers responses indicate that fish are living under stressful environmental conditions. According to multivariate analysis glucose, glutathione S-transferase activity, lipid peroxidation levels and the count of white blood cells are key biomarkers to contribute to discrimination of sites. So, we suggest use those biomarkers in future monitoring of freshwater aquatic systems.