Erythrocytic Nuclear Abnormalities Research Articles

Hematological and genotoxic biomarkers in a natural population of freshwater turtles Trachemys dorbigni (Duméril & Bibron, 1835) (Testudines: Emydidae) living in an urban area in Southern Brazil.

The present study aimed to report the morphometric and hematological indices and genotoxicity of a free-life population of D'Orbigny's slider turtles (Trachemys dorbigni) living in an urban area in Southern Brazil. For that, 16 specimens were randomly captured in an urban canal that receives irregular releases of wastewater. Biometrics and external visual changes were analyzed, such as turtle shell deformities, and the presence of parasites. Blood samples were collected to evaluate the hematological profile and the presence of micronuclei and other erythrocyte nuclear abnormalities as potential mutagenic and genotoxic effects. Water physicochemical parameters were also measured. Organisms with ectoparasites (31.25%) and small carapace deformations (56.25%) were observed, but maximum carapace length and weight were considered normal for the species according to the literature. The blood profile indicated low hemoglobin and hematocrit and a high number of total leukocytes, particularly eosinophils which characterize parasitic infections. A frequency of 0.12% for the micronucleus was considered basal, but the frequency of other erythrocyte abnormalities was evident, mainly of blebbed nuclei (63.79%), indicating chromosomal damage in the early stage. The results of this study suggest that natural populations of chelonian inhabiting urbanized areas are impacted by anthropogenic activities in the surrounding environment. Furthermore, it provides comprehensive data which can serve as a comparative model for environmental monitoring studies involving turtles.

2,4-D-based herbicide underdoses cause mortality, malformations, and nuclear abnormalities in Physalaemus cuvieri tadpoles

Amphibians are considered bioindicators of the environment due to their high sensitivity and involvement in terrestrial and aquatic ecosystems. In the last two decades, 2,4-D has been one of the most widely used herbicides in Brazil and around the world, as its use has been authorized for genetically modified crops and therefore has been detected in surface and groundwater. Against this background, the aim of this work was to investigate the effects of environmentally relevant concentrations of 2,4-D-based herbicides on survival, malformations, swimming activity, presence of micronuclei and erythrocyte nuclear abnormalities in Physalaemus cuvieri tadpoles. The amphibians were exposed to six concentrations of 2,4-D-based herbicides: 0.0, 4.0, 30.0, 52.5, 75.0, and 100 μg L−1, for 168 h. At concentrations higher than 52.5 μg L−1, significantly increased mortality was observed from 24 h after exposure. At the highest concentration (100 μg L−1), the occurrence of mouth and intestinal malformations was also observed. The occurrence of erythrocyte nuclear abnormalities at concentrations of 30.0, 52.5, 75.0 and 100 μg L−1 and the presence of micronuclei at concentrations of 52.5, 75.0, and 100 μg L−1 were also recorded. These effects of 2,4-D in P. cuvieri indicate that the ecological risk observed at concentrations above 10.35 μg L−1 2,4-D may represent a threat to the health and survival of this species, i.e., exposure to 2,4-D at concentrations already detected in surface waters in the species' range is toxic to P. cuvieri.

Multiple morphophysiological responses of a tropical frog to urbanization conform to the pace-of-life syndrome.

The Pace-of-Life syndrome proposes that behavioural, physiological and immune characteristics vary along a slow-fast gradient. Urbanization poses several physiological challenges to organisms. However, little is known about how the health status of frogs is affected by urbanization in the Tropics, which have a faster and more recent urbanization than the northern hemisphere. Here, we analysed a suite of physiological variables that reflect whole organism health, reproduction, metabolic and circulatory physiology and leukocyte responses in Leptodactylus podicipinus. Specifically, we tested how leukocyte profile, erythrocyte morphometrics and germ cell density, as well as somatic indices and erythrocyte nuclear abnormalities differ throughout the adult life span between urban and rural populations. We used Phenotypic Trajectory Analysis to test the effect of age and site on each of the multivariate data sets; and a Generalised Linear Model to test the effect of site and age on nuclear abnormalities. Somatic indices, erythrocyte nuclear abnormalities, erythrocyte morphometrics and leukocyte profile differed between populations, but less so for germ cell density. We found a large effect of site on nuclear abnormalities, with urban frogs having twice as many abnormalities as rural frogs. Our results suggest that urban frogs have a faster pace of life, but the response of phenotypic compartments is not fully concerted.

Exposure to microcrystallized cellulose affects the health of tadpoles and sheds light on the threat these materials pose to amphibians

The increasing use of cellulose-based materials (CBMs) has provided beneficial applications in different sectors. However, its release into environments may represent an ecological risk, therefore demanding that ecotoxicological studies be conducted to understand the risks (current and future) of CBM pollution. Thus, we evaluated the possible effects of microcrystalline cellulose (CMs) in Physalaemus cuvieri tadpoles. After seven days of exposure to CMs (at 58.29 and 100 mg/L), the animals were subjected to behavioral evaluation, and different biomarkers (biometric and biochemical) were evaluated. Although our data do not point to a neurotoxic effect of CMs (inferred by the absence of behavioral changes and changes in AChE and BChE activity), animals exposed to CMs showed differences in body condition. Furthermore, we noticed an increase in the frequency of erythrocyte nuclear abnormalities and DNA damage, which were correlated with the ingestion of CMs. We noticed that the antioxidant activity of tadpoles exposed to CMs (inferred by SOD, CAT, and DPPH radical scavenging activity) was insufficient to control the increase in ROS and MDA production. Furthermore, exposure to CMs induced a predominant Th2-specific immune response, marked by suppressed IFN-γ and increased IL-10 levels, with a consequent reduction in NO levels. Principal component analysis and IBRv-2 indicate, in general, a primarily more toxic response to animals exposed to the highest CM concentration. Therefore, our study evidence that CMs affect the health of P. cuvieri tadpoles and sheds light on the threat these materials pose to amphibians.

A Deep Survey of Fish Health for the Recognition of Useful Biomarkers to Monitor Water Pollution

The aim of the present study was to evaluate the wild freshwater fish health status using a vast array of biomarkers as predictive factors of pollutant exposure. The European eel (Anguilla anguilla) and brown trout (Salmo trutta fario), resident in rivers with different degrees of pollution in the South of Italy (Picentino River with good environmental quality and Tusciano River with low environmental quality), were examined using biometric parameters, histopathological and immunohistochemical biomarkers to evaluate the health status and a possible correlation with the water quality. Several alterations identified in the liver positively correlated with water and soil pollutants: hemorrhage (p ≤ 0.05), cytoplasmic vacuolization (p ≤ 0.01), hemosiderosis (p ≤ 0.05), irregular arrangement of hepatocytes (p ≤ 0.01), lipid accumulation (p ≤ 0.05), necrosis (p ≤ 0.01), cellular hyperplasia (p ≤ 0.05), leukocyte infiltration (p ≤ 0.01) and melanomacrophages centers (MMC) (p ≤ 0.01). In the spleen, only hemosiderosis correlated with water and soil pollutants (p ≤ 0.05). The inflammatory biomarker tumor necrosis factor α (TNFα) and ciclooxigenase 2 (COX2) responded to the environmental pollution, as well as the oxidative stress biomarkers superoxide dismutase (SOD2) and 8-Hydroxy-2′-deoxyguanosine (8-OHdG). Erythrocytic nuclear abnormalities and erythrocytic cellular abnormalities were found to be significantly higher in the blood of both the European eel (p < 0.0001) and brown trout (p < 0.001) in the Tusciano River compared with the Picentino River. Taken together, these results outline the need to increase the number of suitable biomarkers to assess fish health and reinforce the importance of employing additional biomarkers in biomonitoring programs that can be applied to evaluate water quality and in environmental assessment around the world.

Per- and polyfluoroalkyl substances (PFAS) in little penguins and associations with urbanisation and health parameters

Per- and Polyfluoroalkyl substances (PFAS) are increasingly detected in wildlife and present concerning and unknown health risks. While there is a growing body of literature describing PFAS in seabird species, knowledge from temperate Southern Hemisphere regions is lacking. Little penguins (Eudyptula minor) can nest and forage within heavily urbanised coastal environments and hence may be at risk of exposure to pollutants. We analysed scat contaminated nesting soils (n = 50) from 17 colonies in lutruwita/Tasmania for 16 PFAS, plasma samples (n = 45) from nine colonies, and three eggs for 49 PFAS. We detected 14 PFAS across the sample types, with perfluorooctanesulfonic acid (PFOS) and perfluorohexanesulfonic acid (PFHxS) most commonly detected. Mean concentration of PFOS in plasma was 2.56 ± 4.3 ng/mL (<LOD–22.3), soil was 2.92 ± 5.39 μg/kg (<LOD–29.8) and eggs were 11.89 ± 11.30 μg/kg (0.27–22.84). Mean concentration of PFHxS in plasma was 0.9 ± 0.84 ng/mL (<LOD–5.6), soil was 11.4 ± 39.18 μg/kg (<LOD–213.3) and eggs were 0.27 ± 0.11 μg/kg (<LOD–0.38). Urbanisation of breeding sites was positively associated with PFOS and PFHxS concentrations in soils and blood. Phenology appeared to be an important predictor of PFHxS load, with moulting birds having lower levels compared to non-moulting birds. We investigated PFAS associations with health parameters and found that haematocrit and total plasma proteins in females were negatively associated with PFOS and positively associated in males. The number of erythrocytic nuclear abnormalities in males was positively associated with PFOS. Findings are the first to document the presence of PFAS in this resident seabird nesting and foraging close to anthropogenically modified habitats. We demonstrate that even in low concentrations, PFAS may have detectable relationships with penguin health. Ongoing research is needed to investigate foraging ecology and trophic bioaccumulation pathways, as well as understand sublethal consequences from exposure.

Erythrocytic nuclear abnormalities in Cyprinus carpio L. cultivated in water bodies of agroecosystems

Abstract Agrochemicals contaminate water bodies in agroecosystems, which affects the health of fishes. We assessed erythrocytic nuclear abnormalities (ENA) in Cyprinus carpio reared in uncontaminated and contaminated water bodies. We recorded micronucleus, notched nucleus, lobed nucleus, kidney bean-shaped nucleus, bi-nucleated, blebbed nucleus, karyopyknosis, nuclear shift, ooze out nucleus, and elongated nucleus at higher incidences in fish reared in the contaminated site. We observed a high percent incidence of ENA and micronucleus in fish from the contaminated site. The analysis of heavy metals with atomic absorption spectroscopy revealed that all heavy metals (Mn, Zn, Ni, Pb, Cd, Fe) were high in the contaminated site except for Cu in water and Cu and Cr in sediments. Correlation matrix analysis showed that total ENA increased with Cu (r = 0.996, p &lt; 0.0001), Mn (r = 0.942, p &lt; 0.0001), Zn (r = 0.517, p = 0.07), and Cr (r = 0.997, p = 0.0001). Compared to the uncontaminated site, the incidence of micronuclei was 20 times higher in the fish from the contaminated site, and this was influenced by the Cu, Mn, Zn, and Cr contents in the sediments. This study indicated that the heavy metal contamination of a water body located in an agroecosystem contributed to nuclear abnormalities in C. carpio.

The Effects of Acute Bisphenol A Toxicity on the Hematological Parameters, Hematopoiesis, and Kidney Histology of Zebrafish (Danio rerio)

In this study, the results of evaluating the acute toxicity of Bisphenol A on Danio rerio are presented, encompassing peripheral blood parameters, the composition of hematopoietic cells of erythroid and myeloid lines in the head kidney, and data from histological studies. The LC50 values of Bisphenol A for adult zebrafish individuals for 12, 24, and 48–96 h were determined, which were 18.04, 7.55, and 6.22 mg/L, respectively. The study includes data on the morphology and quantitative frequency of specific cells in the hematopoietic tissue of the head kidney, along with the consideration of adaptive mechanisms in hematopoiesis under BPA exposure. The application of polynomial regression analysis to reveal the concentration–effect relationship for some hematological and histological parameters was demonstrated. Significant increases in the frequency of erythrocyte nuclear abnormalities were observed at BPA concentrations of 6 and 8 mg/L, which indicates a genotoxic effect. BPA’s impact on fish peripheral blood parameters manifested as an increase in the number of erythrocytes (RBC) and immature erythrocytes, as well as a decrease in the number of lymphocytes. The most notable pathological changes in the head kidney’s hematopoietic tissue included circulatory disturbances and the formation of inflammation/degradation foci, as confirmed by histopathologic indices. At BPA concentrations of 2 and 4 mg/L, the observed changes were compensated for by hematopoietic adaptation mechanisms; however, at concentrations of 6 and 8 mg/L, acute systemic toxicity was evident.

Mortality and toxicity of a commercial formulation of cypermethrin in Physalaemus gracilis tadpoles

This study evaluated the lethal, sublethal, and toxic of a commercial formulation of cypermethrin in the anuran species Physalaemus gracilis. In the acute test, concentrations of 100–800 μg L−1 were tested over 96 h. In the chronic test, cypermethrin concentrations recorded in nature (1, 3, 6, and 20 μg L−1) were tested for mortality and then used for the micronucleus test and erythrocyte nuclear abnormalities over a 7-days period. The LC50 determined for P. gracilis for the commercial cypermethrin formulation was 273.41 μg L−1. In the chronic test, a mortality of more than 50% was observed at the highest concentration (20 μg L−1), as it caused half of the tadpoles studied to die. The micronucleus test showed significant results at concentrations of 6 and 20 μg L−1 and recorded the presence of several nuclear abnormalities, indicating the genotoxic potential of the commercial cypermethrin formulation for P. gracilis. Cypermethrin presented a high risk to the species, indicating that it has the potential to cause several problems in the short and long term and to affect the dynamics of this ecosystem. Therefore, it can be concluded that the commercial formulation of cypermethrin had toxicological effects on P. gracilis.

Chronic Degradation of Seasonally Dry Tropical Forests Increases the Incidence of Genotoxicity in Birds.

Multiple studies have shown that exposure to pollutants can increase genotoxic damage in different taxa. However, to our knowledge, the effects of environmental stress have been explored little. In certain stressful ecosystems, such as seasonally dry tropical forests, the combined effects of anthropogenic activities and ongoing global changes can cause an increase in environmental stresses, in turn, may trigger physiological and genetic effects on biodiversity. The present aims to assess changes in the prevalence of genotoxic damage in birds within three states of forest degradation in the Tumbesian Region of Western Ecuador. We used blood samples from 50 bird species to determine the frequency of micronucleus and nuclear abnormalities in erythrocytes. Our results revealed a significant impact of forest degradation on the occurrence probability of micronucleus and nuclear abnormalities at the community level. Localities with higher levels of degradation exhibited higher levels of abnormalities. However, when analyzing the dominant species, we found contrasting responses. While Lepidocolaptes souleyetii showed a reduction in the proportion of nuclear abnormalities from the natural to shrub-dominated localities Troglodytes aedon and Polioptila plumbea showed an increase for semi-natural and shrub-dominated respectively. We concluded that the degradation process of these tropical forests increases the stress of bird community generating genotoxic damage. Bird responses seem to be species-specific, which could explain the differences in changes in bird composition reported in other studies.

Acute toxicity and genotoxicity of the S-metolachlor-based herbicide Dual Gold® on Leptodactylus luctator (Hudson, 1892) tadpoles (Anura: Leptodactylidae).

Herbicides used in agriculture and their metabolites are frequently detected in surface water bodies, where they can persist and cause adverse effects on aquatic organisms. The aim of this study was to evaluate the acute toxicity and genotoxic effects of the S-metolachlor (SM)-based herbicide Dual Gold® (DG®), on Leptodactylus luctator tadpoles (Anura: Leptodactylidae). To assess the toxicity of the herbicide, including the median lethal concentration (LC50) at 24h, the no-observed-effect concentration (NOEC), and the lowest-observed-effect concentration (LOEC), tadpoles were exposed to five nominal concentrations of DG® (5.0, 6.2, 7.8, 9.8, and 12.2 mg/L), and to dechlorinated water as a negative control (NC). The LC5024h of DG® was 7.0 mg/L, the NOEC was 5.0 mg/L and the LOEC=6.2 mg/L. L. luctator tadpoles were sensitive to the herbicide, reaching 100% mortality after 24 h of exposure to the highest concentration tested (12.2 mg/L). To evaluate the potential genotoxicity of the herbicide, the frequencies of micronuclei (MN) and other erythrocyte nuclear abnormalities (ENA) were determined in larvae exposed to three nominal concentrations of DG® (1.0, 5.0, and 6.2 mg/L) for 48 and 96 h. The frequencies of MN and ENA were compared with a positive control (40 mg/L of Cyclophosphamide) and a negative control. The frequencies of MN and ENA in the erythrocytes of tadpoles exposed to the test concentrations of DG® and Cyclophosphamide were significantly higher than in the negative control group at both 48 and 96 h (with the only exception of MN at 1.0 mg/L at 48 h). Our results confirm the genotoxic and cytotoxic effects of this widely used herbicide in agriculture, a fact that represents a potential risk to amphibians that develop in ponds associated with or immersed in agroecosystems.

The Use of Multiple Biomarkers to Assess the Health of Anuran Amphibians in the Brazilian Cerrado Savanna: An Ecotoxicological Approach.

Changes in the natural landscape and the indiscriminate use of pesticides can have a major impact on aquatic environments and have contributed to the worldwide decline of amphibian populations. In the present study, we sampled tadpoles of three anuran amphibians (Boana albopunctata, Physalaemus cuvieri, and Dendropsophus minutus) from ponds in six different agricultural landscapes of the Brazilian Cerrado savanna and evaluated whether and to what extent genotoxic and mutagenic damage was related to land use (the amount of forest and agricultural remnants, and related physicochemical factors) and the presence of pesticides in the water of the study ponds. We also evaluated the hepatotoxicity in P. cuvieri, which was the most abundant species at five of the six sampling points. Clomazone and atrazine were the most common pesticides found in the ponds. The B. albopunctata and P. cuvieri tadpoles presented similar patterns of DNA damage among the sampling points. The least DNA damage was found in the D. minutus tadpoles, although this species was present in only one of the study ponds. More binucleated and anucleated cells were observed in B. albopunctata, but there was no significant variation among species in terms of the number of micronuclei or other erythrocytic nuclear abnormalities. Land use and physicochemical factors did not explain the variation in the DNA damage observed in the three anurans. The hepatotoxicity analyses of P. cuvieri revealed the presence of a series of alterations, including the enlargement of the sinusoids, vacuolization of the hepatocytes, the infiltration of inflammatory cells, hepatic steatosis, and dilation of the blood vessels. The interaction between physicochemical factors and the biomarkers analyzed in the present study is complex. In particular, it will be important to better elucidate which factors are contributing, either directly or indirectly, to the decline of anuran amphibian populations, especially in threatened biomes, such as the Brazilian Cerrado. In this case, we would encourage further in situ studies that assess the ecotoxicology of the landscape, together with the systematic monitoring of aquatic environments, to guarantee the long-term integrity of amphibian populations, and those of other organisms that play an essential functional role in the ecosystem. Environ Toxicol Chem 2023;00:1-18. © 2023 SETAC.

Mutagenicity, hepatotoxicity, and neurotoxicity of glyphosate and fipronil commercial formulations in Amazon turtles neonates (Podocnemis expansa)

Pesticides are considered one of the main causes of the population decline of reptiles worldwide, with freshwater turtles being particularly susceptible to aquatic contamination. In this context, we investigated the potential mutagenic, hepatotoxic, and neurotoxic effects in neonates of Podocnemis expansa exposed to substrate contaminated with different concentrations of glyphosate and/or fipronil during embryonic development. Eggs collected from the natural environment were artificially incubated in sand moistened with pure water, water added with glyphosate Atar 48® at concentrations of 65 and 6500 μg/L (groups G1 and G2, respectively), water added with fipronil Regent® 800WG at 4 and 400 μg/L (groups F1 and F2, respectively) and, water added with the combination of 65 μg/L glyphosate and 4 μg/L fipronil or with 6500 μg/L glyphosate and 400 μg/L fipronil (groups GF1 and GF2, respectively). For mutagenicity analysis, we evaluated the frequency of micronuclei (MN) and other erythrocyte nuclear abnormalities (ENAs), while for evaluation of hepatotoxicity and neurotoxicity, livers and encephalon were analyzed for histopathological alterations. Exposure to pesticides, alone or in combination, increased the frequency of erythrocyte nuclear abnormalities, particularly blebbed nuclei, moved nuclei, and notched nuclei. Individuals exposed to fipronil exhibited congestion and inflammatory infiltrate in their liver tissue, while, in the encephalon, congestion, and necrosis were present. Our study confirms that the incubation of eggs in substrate polluted with glyphosate and fipronil causes histopathological damage and mutagenic alteration in P. expansa, highlighting the importance of using different biomarkers to evaluate the ecotoxicological effects of these pesticides, especially in oviparous animals.

Erythrocytic Nuclear Abnormalities and Leucocyte Profiles of Asian Seabass (Lates calcarifer) Exposed to Polluted Seawater

Close to industrial activity and a major seaport in the Gulf of Thailand, Sichang Island has frequently suffered from pollution and oil spills. However, the environmental health status of the surrounding waters is relatively unknown. Between December 2017 and January 2018, we reared ninety Asian seabass (Lates calcarifer) in cage nets off the island and sampled blood to investigate nuclear abnormalities in erythrocytes and build leucocyte profiles. The fish were purchased locally and reared in locations where oil spills have been recorded. Environmental parameters were within the range of standard values. Erythrocyte nuclei had segmented, reniform and notched abnormalities; however, they dramatically varied after exposure. At the end of the three-month field experiment, very few micronuclei had been observed. Observed leucocytes were neutrophils, lymphocytes, and monocytes. The percentage of lymphocytes increased considerably, reaching a post-exposure peak at three months. Our data suggest that erythrocytic nuclear abnormalities of Asian seabass could be used as an early warning of toxic pollutants in the marine environment, and as a baseline environmental health indicator for Sichang Island and the surrounding area.

Integrated Use of Bioaccumulation, Genotoxic, and Haematological Endpoints to Assess the Effect of Water Remediation Strategies on Fish Health: A Complementary Study

Biosorption successfully remediates saline water contaminated with legacy contaminants, but its effects on the health of marine organisms remain unclear. Therefore, our aim was to address this knowledge gap with data on the accumulation ability, as well as the cytogenetic and biochemical effects in turbot (Scophthalmus maximus). To this end, we exposed turbot for seven days to a mixture of remediated metals (Rem treatments: Cd, Hg, and Pb), with and without the presence of nanoparticles (NP), and compared them with the maximum allowable concentrations (MAC treatment) for effluent discharges. We determined the metal accumulation in the blood and kidney and evaluated haematological changes (red blood cell count, haemoglobin, and mean cell haemoglobin (MCH)) and genotoxicity (erythrocytic nuclear abnormalities assay) in the blood. The results showed that remediation with non-living macroalgae significantly reduced the metallic blood and kidney burdens in the Rem treatments. Furthermore, no genotoxic potential occurred in the Rem and MAC treatments in parallel with the reduction in MCH levels in the Rem treatments, which would reflect hematopoietic disturbances in the MAC. Our results validate biosorption remediation as we achieved a considerable reduction in metal loads while maintaining the health status of fish, highlighting the importance of testing water remediation methods in the biota.

Exposure of adult zebrafish (Danio rerio) to SARS-CoV-2 at predicted environmentally relevant concentrations: Outspreading warns about ecotoxicological risks to freshwater fish

While the multifaceted social, economic, and public health impacts associated with the COVID-19 pandemic are known, little is known about its effects on non-target aquatic ecosystems and organisms. Thus, we aimed to evaluate the potential ecotoxicity of SARS-CoV-2 lysate protein (SARS.CoV2/SP02.2020.HIAE.Br) in adult zebrafish (Danio rerio) at predicted environmentally relevant concentrations (0.742 and 2.226 pg/L), by 30 days. Although our data did not show locomotor alterations or anxiety-like or/and anxiolytic-like behavior, we noticed that exposure to SARS-CoV-2 negatively affected habituation memory and social aggregation of animals in response to a potential aquatic predator (Geophagus brasiliensis). An increased frequency of erythrocyte nuclear abnormalities was also observed in animals exposed to SARS-CoV-2. Furthermore, our data suggest that such changes were associated with a redox imbalance [↑ROS (reactive oxygen species), ↑H2O2 (hydrogen peroxide), ↓SOD (superoxide dismutase), and ↓CAT (catalase)], cholinesterasic effect [↑AChE (acetylcholinesterase) activity], as well as the induction of an inflammatory immune response [↑NO (nitric oxide), ↑IFN-γ (interferon-gamma), and ↓IL-10 (interleukin-10)]. For some biomarkers, we noticed that the response of the animals to the treatments was not concentration-dependent. However, principal component analysis (PCA) and the “Integrated Biomarker Response” index (IBRv2) indicated a more prominent ecotoxicity of SARS-CoV-2 at 2.226 pg/L. Therefore, our study advances knowledge about the ecotoxicological potential of SARS-CoV-2 and reinforces the presumption that the COVID-19 pandemic has negative implications beyond its economic, social, and public health impacts.

Description and quantification of micronucleus and nuclear abnormalities in erythrocytes of the sentinel green turtle (Chelonia mydas) with fluorescence microscopy.

Contamination in marine ecosystems is of the most critical threats to marine turtles. The identification of useful biomarkers to detect and monitor the physiological and clinical effects of pollutants on these populations will allow early detection of alterations (e.g., mutagenic damages) that could risk their viability or favor the development of diseases, thus threatening the biodiversity of these ecosystems and human population. This study is aimed at describing and quantifying nuclear anomalies in peripheral blood erythrocytes of green turtles (Chelonia mydas) from three distinct foraging areas in Mexico (Akumal, Xcalak, and Punta Herrero). We developed a novel morphological index that could be used as a biomarker to identify abnormal nuclei in peripheral blood erythrocytes. Here we describe for the first time in C. mydas, with a species-specific staining protocol, distinct nuclear abnormalities such as blebbed, lobed, notched, eight shape nuclei, and binucleated cells. These nuclear abnormalities were present in >90 % of the subjects (n=30). Moreover, 50 % of the organisms presented erythrocytes with micronuclei. The number of nuclear abnormalities did not correlate with size of the green turtles or differ between sites, or health status. We found a higher frequency of green turtles with nuclear abnormalities in the southern region (Punta Herrero and Xcalak) with the highest frequency of micronucleus and buds. The former could be associated to the constant exposure to chemical pollutants of oceanographic origin in the southern coast of Quintana Roo. Furthermore, the increasing anthropogenic pollution in Akumal could also explain the highest variability in the number of nuclear abnormalities presented in resident individuals. We propose that a long-term monitoring programs of green turtle populations in the Mexican Caribbean that include a micronucleus test could be a useful to determine possible mutagenic damage in these animals.

Oxidative Stress in a Wild Population of the Freshwater Fish Hyphessobrycon Luetkenii Chronically Exposed to a Copper Mining Area: New Insights into Copper Toxicology.

In this study, we examine markers of oxidative stress in the tetra Hyphessobrycon luetkenii collected from two locations in the copper contaminated João Dias creek (southern Brazil). Also, specimens were translocated from a clean reference section of the creek to a polluted stretch and vice-versa. Fish were held at in submerged cages for 96h and then sacrificed. Nuclear abnormalities in erythrocytes and total antioxidant capacity, lipid peroxidation and protein carbonylation in gills, brain, liver and muscle displayed similar trends in both groups. Lipid peroxidation increased in all tissues of individuals translocated to the polluted site but only in liver and muscle of those translocated to the reference site. Increased protein carbonylation was also observed in gills of individuals translocated to the reference location. These results suggest similar oxidative stress among fish from the reference and polluted locations and that long-term metals exposure may require adaptations toward oxidative stress responses.

Potential ecotoxicity of substrate-enriched zinc oxide nanoparticles to Physalaemus cuvieri tadpoles

Although the ecotoxicological effects of ZnO nanoparticles (ZnO NPs) have already been reported in different taxa, little is known about their impacts on amphibians. Thus, we aimed to evaluate the potential effects of exposure of Physalaemus cuvieri tadpoles to substrates enriched with ZnO NPs (and with its ionic counterpart, Zn+2, ZnCl2 – both at 100 mg/kg) previously used in the cultivation of Panicum maximum (Guinea grass). We showed that although exposure for 21 days did not impact the survival, growth, and development of tadpoles, we noted an increase in the frequency of erythrocyte nuclear abnormalities in the “ZnCl2” and “ZnONP” groups, which was associated with suppression of antioxidant activity in the animals (inferred by SOD and CAT activity and DPPH free radical scavenging capacity). In the tadpoles of the “ZnONP” group, we also noticed a reduction in creatinine and bilirubin levels, alpha-amylase activity, and an increase in alkaline phosphatase activity. But the treatments did not alter the activity of the enzymes lactate dehydrogenase and gamma-glutamyl-transferase and total protein and carbohydrate levels. On the other hand, we report a cholinesterase and hypotriglyceridemic effect in the “ZnCl2” and “ZnONP” groups. Zn bioaccumulation in animals, from ZnO NPs, from Zn+2 released from them, or both, has been associated with causing these changes. Finally, principal component analysis (PCA) and the values of the “Integrated Biomarker Response” index revealed that the exposure of animals to substrates enriched with ZnO NPs caused more pronounced effects than those attributed to its ionic counterpart. Therefore, our study reinforces the need to consider the environmental risks of using these nanomaterials for agricultural purposes for amphibians.

Brodifacoum Levels and Biomarkers in Coastal Fish Species following a Rodent Eradication in an Italian Marine Protected Area: Preliminary Results.

Brodifacoum is the most common rodenticide used for the eradication of invasive rodents from islands. It blocks the vitamin K cycle, resulting in hemorrhages in target mammals. Non-target species may be incidentally exposed to brodifacoum, including marine species. A case study conducted on the Italian Marine Protected Area of Tavolara Island was reported after a rodent eradication using the aerial broadcast of a brodifacoum pellet. Brodifacoum presence and effects on non-target marine organisms were investigated. Different fish species were sampled, and a set of analyses was conducted to determine vitamin K and vitamin K epoxide reductase concentrations, prothrombin time, and erythrocytic nuclear abnormalities (ENA) assay. In all the examined organisms, brodifacoum was not detected. The results obtained showed differences in vitamin K and vitamin K epoxide concentrations among the samples studied, with a positive correlation for three species between vitamin K, vitamin K epoxide, and fish weight. The prothrombin time assay showed a good blood clotting capacity in the fish. Higher abnormality values were recorded for four species. The results of this study suggest that it is possible to hypothesize that the sampled fish were not likely to have been exposed to brodifacoum and that consequently there are no negative issues concerning human consumption.