Genotoxicity Biomarkers Research Articles

Micronuclei in embryos of eight seabird species in northwestern Mexico: A biomarker of exposure to coastal pollution?

The micronucleus (MN) test may be used to evaluate genome instability in birds and the potential of different species to function as biomarkers of genotoxicity. However, little is known regarding genome instability in seabird embryos or the instability present among embryonic development stages. Therefore, the present study aimed to describe the frequencies of micronucleated erythrocytes (MNE) and micronucleated polychromatic erythrocytes (MNPCE) in blood samples collected from the embryos of eight seabird species nesting on the coast of Sinaloa, Mexico. An additional description of blood cell maturation along with embryo development during incubation was conducted based on the proportion of polychromatic erythrocytes (PCE), and the potential relationships between metals (Hg and Cd concentrations in egg content) and the MN frequencies in embryo blood were evaluated. The PCE proportion appears to decline as incubation advances (initial stage > intermediate stage > advanced stage), and the values varied between species (Suliformes/Pelecaniformes < Charadriiformes: Laridae), which may be related to differences among incubation periods and reproductive strategies. Interspecific variation in the MNPCE frequency was found in embryos showing advanced development, which could be related to both variations in life-history traits and ecological factors and not Hg or Cd exposure. The genomic instability values in this study are the first to be reported for embryos of seabird species nesting in a subtropical coastal region.

Metals, pesticides, and emerging contaminants on water bodies from agricultural areas and the effects on a native amphibian

In the Paraná River lower basin, an important agro-productive area of Argentina, crop fields and cattle breeding activities are common and may affect water quality. So, the aim of this study was to analyze the impacts of cattle breeding and agricultural activities on a stream from Buenos Aires, through physicochemical parameters (metals, pesticides, and emerging contaminants) and ecotoxicological parameters with Rhinella arenarum larvae, a native amphibian species. Three sites were selected on an ordinary plain stream that goes through agricultural fields and a cattle breeding establishment (upstream –S1-, near –S2- and downstream –S3- the establishment). Physicochemical parameters were measured in situ (in water) and in laboratory (in water and sediment samples: metals, pesticides, ivermectin and oxytetracycline). A semi-static chronic toxicity bioassay (504 h) was performed with water samples, and neurotoxicity, oxidative stress and genotoxicity biomarkers were measured after acute exposure (96 h). According to the index, a degradation in the water quality was observed in all sites. Ivermectin (8.03 mg/kg) and oxytetracycline (1.9 mg/kg) were detected in sediment samples from S2. Pesticides were detected in all sites, mainly in water samples: S1 presented the highest variability (7 residues) and in S3 AMPA, glyphosate and acetochlor concentrations were higher (10.3, 22.4 and 23.8 μg/L). Also, all sites significantly produced lethality at chronic exposure. Lethality at 504h was 40% for S1, 56.66% for S2 and 93.33% for S3. At acute exposure, the oxidative stress biomarkers were altered on R. arenarum larvae exposed to all sites and the neurotoxicity biomarkers were altered on larvae exposed to S1 and S3. Water quality was severely degraded by the surrounding agricultural and cattle breeding activities, which may represent a threat to the ecosystems.

Association of NEF2L2 Rs35652124 Polymorphism with Nrf2 Induction and Genotoxic Stress Biomarkers in Autism.

Increased oxidative/genotoxic stress is known to impact the pathophysiology of ASD (autism spectrum disorder). Clinical studies, however, reported limited, heterogeneous but promising responses to treatment with antioxidant remedies. We determined whether the functional polymorphism of the Nrf2 gene, master regulator of anti-oxidant adaptive reactions to genotoxic stress, links to the genotoxic stress responses and to an in vitro effect of a NRF2 inductor in ASD children. Oxidative stress biomarkers, adaptive responses to genotoxic/oxidative stress, levels of master antioxidant regulator Nrf2 and its active form pNrf2 before and after inducing by dimethyl fumarate (DMF), and promotor rs35652124 polymorphism of NFE2L2 gene encoding Nrf2 were studied in children with ASD (n = 179). Controls included healthy adults (n = 101). Adaptive responses to genotoxicity as indicated by H2AX and cytoprotection by NRF2 contents positively correlated in ASD children with a Spearman coefficient of R = 0.479 in T+, but not CC genotypes. ASD children with NRF2 rs35652124 CC genotype demonstrated significantly higher H2AX content (0.652 vs. 0.499 in T+) and pNrf2 induction by DMF, lowered 8-oxo-dG concentration in plasma and higher cfDNA/plasma nuclease activity ratio. Our pilot findings suggest that in ASD children the NEF2L2 rs35652124 polymorphism impacts adaptive responses that may potentially link to ASD severity. Our data warrant further studies to reveal the potential for NEF2L2 genotype-specific and age-dependent repurposing of DMF and/or other NRF2-inducing drugs.

In vitro genotoxicity potential investigation of 7 oxy-PAHs.

Air pollutants include many compounds among them oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs). As they are suspected to generate DNA damage and mutagenicity, an understanding of their mode of action could highlight a carcinogenic potential risk in exposed population. In this article, a prospective study on seven oxy-PAHs selected in terms of occurrence in the environment was conducted on mutagenicity, genotoxicity, and cytotoxicity potentials using in vitro assays including Ames test on five strains, kinetic analysis of cytotoxicity and apoptosis, phosphorylation of histone H2AX, and p53 induction assays on human lung cell line BEAS-2B. Ames test demonstrated that mutagenicity pattern depended on the oxy-PAH tested. Except for BAQ, all oxy-PAHs tested gave mutagenic effect, in the absence and/or in the presence of metabolic activation (S9 fraction). At 24 h of exposure, the majority of oxy-PAHs induced γ-H2AX in BEAS-2B cells and/or phosphorylation of p53 at serine 15 and cell death at highest tested concentrations. Although 9,10-AQ and B[b]FO were mutagenic in bacteria, they failed to induce any of the other genotoxicity biomarkers. In comparison with the benzo[a]pyrene, all oxy-PAHs were less potent in terms of genotoxic potential at the same concentration. These results highlighted the genotoxic and mutagenic potential of these oxy-PAHs and provide preliminary information concerning their possible mechanism of action for toxicity, contributing to a better evaluation of the real associated health risks for human and environment.

Sub-Lethal Effects Of Chlorpyrifos On Geno-Neurotoxic Profiles In Freshwater Fish Cyprinus Carpio(L.)

Pesticide toxicity is a global concern and chlorpyrifos is one of the most widely used organophosphate agrochemicals,it is frequently detected in surface ground waters of India. Chlorpyrifos(CPF) LC50 – 96 h was calculated as 0.318 mg/l using probit analysis based on LC50 value two sublethal concentrations of 1/5th of LC50 (0.0636 mg/l) and 1/10th of LC50 (0.0318 mg/l) were determined. The influence of CPF on the acetylcholinesterase (AChE) activity and Ach content and the Genotoxicity of Cyprinus carpio was evaluated. Results confer that significant inhibition in the AchE activity in brain tissue resulting in a build-up of Ach at the neuronal synapse resulting in a decrease of cholinergic transmission finally impairs the neurophysiological activity and ultimately result in the death of fish although studies on genotoxic effects of CPF were carried out to examine the prevalence of nuclear abnormalities in freshwater fish Cyprinus carpio using Comet and MN assay in blood erythrocytes and gill cells as percentile tail DNA damage as comets and micronuclei and other nuclear anomalies were scored in erythrocytes of Cyprinus carpio it was found that significant effect was observed for both concentrations and periods of exposure in the treated fishes. The highest DNA damage and increased micronuclei frequency and inhibitory AchE activity in Cyprinus carpio were found to be high on the highest sublethal 1/5th of LC50 followed by Sublethal 1/10th of LC50 concentration. Overall findings imply that AchE and genotoxic biomarkers may be useful diagnostic tools for chlorpyrifos poisoning in biomonitoring programs.

Salvinia molesta phytoremediation capacity as a nature-based solution to prevent harmful effects and accumulation of ciprofloxacin in Neotropical catfish.

Phytoremediation has been a potential solution for the removal of pharmaceuticals from water. Here, we evaluated the toxicological safety of ciprofloxacin-contaminated water treated by 96h with Salvinia molesta. The Neotropical catfish Rhamdia quelen was used as a model, and the potential of the phytoremediation technique for mitigating the drug accumulation in the fishes was also studied. Fish exposed to Cipro (1 and 10µg·L-1) in untreated water showed toxic responses (alteration of hematological, genotoxicity, biochemical, and histopathological biomarkers) and accumulated Cipro in their muscles at concentrations high for human consumption (target hazardous quotient > 1). Fish exposed to water treated with S. molesta showed no toxic effect and no accumulation of Cipro in their tissues. This must be related to the fact that S. molesta removed up to 97% of Cipro from the water. The decrease in Cipro concentrations after water treatment with S. molesta not only prevented the toxic effects of Cipro on R. quelen fish but also prevented the antimicrobial accumulation in fish flesh, favouring safe consumption by humans. For the very first time, we showed the potential of phytoremediation as an efficiently nature-based solution to prevent environmental toxicological effects of antimicrobials to nontarget organisms such as fish and humans. The use of S. molesta for Cipro-removal from water is a green technology to be considered in the combat against antimicrobial resistance.

Using native fish in eco-genotoxic assessment of heavy metal contamination pollution arising from nearby large Brazilian rivers

ABSTRACT Worldwide water quality has declined progressively due to continuous pollution of aquatic resources by agrochemicals in particular heavy metals. Fish genotoxicity biomarkers are vital to identify and complement chemical parameters for determining environmental risk of adverse effects. Therefore, it was of interest to examine the eco-genotoxicity attributed to water pollution over different stream sections of Brazilian rivers by using Cichlasoma paranaense (Teleostei: Cichlidae), a neotropical freshwater cichlid fish, as a biological model. Chemical analysis of water and sediments collected from different Brazilian rivers sites demonstrated contamination by metals. Cichlasoma paranaense were collected at a reference location (a permanent water preservation area), maintained in the lab under standard conditions (controlled temperature, lighting, daily feeding, and constant aeration) and exposed to environmental samples of water and sediments. Subsequently, micronucleus (MN) and nuclear abnormalities (NA) frequencies were assessed in erythrocytes obtained from the caudal and gill regions. The highest concentrations of Cu were found in samples from river sites with forest fragmentation attributed to intensive agriculture practices. Similarly, exposure of fish to samples from agricultural areas induced significantly higher number of genotoxic effects. There was no marked difference between the tissues (tail and gill) regarding the observed frequencies of MN and NA. Thus C. paranaense fish served as a reliable model for detecting genotoxic effects, especially when water samples were collected near the discharge of agrochemicals. Evidence indicates that this method be considered for other global river sites which are also exposed to agrochemicals discharges containing Cu.

Association of MTHFR C677T polymorphism with risk of preterm birth in Indian mothers: a case–control study

BackgroundSince MTHFR is the key enzyme in folate metabolism, its reduction can lead to hyperhomocysteinemia, which can have a negative impact on pregnancy outcome. Moreover, MTHFR polymorphism has also been linked with oxidative stress and genotoxicity. Identifying its ethnicity-specific association can help to reduce the incidence of preterm birth (PTB). Material and methods: Age-matched preterm birth mothers (< 37 weeks) and full-term mothers (> 37 weeks) were carefully selected for the study. The polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method was adopted to analyse MTHFR C677T polymorphism. Oxidative stress (OS) analysis was performed by measuring the levels of antioxidants (superoxide dismutase (SOD) and catalase (CAT)) and OS damage markers (lipid peroxidation (LPO), total protein). Genotoxicity was confirmed by the cytokinesis-block micronucleus cytome (CBMN-Cyt) assay. The statistical analysis was performed by Student’s t test, chi-square test, and one-way ANOVA. The relevant risk of premature delivery was represented by odds ratios (ORs) with 95% confidence intervals (95% CIs).ResultsThe MTHFR polymorphism showed statistical significance for PTB outcome with CT and TT genotype frequencies at p < 0.01 and p < 0.05, respectively, between cases and controls. Within the PTB categories (extreme-, very-, moderate-PTB), TT genotype showed statistical significance at (p < 0.05), while CT genotype remained insignificant. Also, statistically high oxidative stress and DNA damage were observed in cases compared to controls for all genotypes. Furthermore, the T allele of the MTHFR gene was found to be linked with significantly increased OS and DNA damage on comparison within the groups.ConclusionsThis study confirms the MTHFR C677T polymorphism, oxidative stress, and genotoxicity biomarkers are associated with the PTB outcome. Analysis of these biomarkers during pregnancy can be of clinical significance.

Systematization of research on the incidence of pesticides in people, use of biomarkers

Currently the use of pesticides in agriculture has expanded in the search for greater productivity. These products can harm people's health in various ways. These effects can be captured through the use of genotoxicity biomarkers. The objective of this research is to systematize studies on biomarkers of genotoxicity of people exposed to pesticides in South America. The PRISMA method was applied to determine the studies to be analyzed. 15 documents met the inclusion criteria. Among the adverse health effects perceived in studies are neurological, respiratory, dermatological and endocrine disorders, as well as an increased risk of cancer. The main biomarkers identified are the comet assay, the cytokinesis blockade micronucleus assay, and the buccal cytoma micronucleus assay. Polymerase chain reaction, chromosomal aberrations, flow cytometry, and fluorescence in situ hybridization were also taken into account. Limitations were determined by biomarker. The usefulness of using multiple biomarkers is highlighted for a more complete and precise evaluation of pesticide exposure and genotoxic damage in agricultural workers in South America. The establishment of protective measures for workers against the use of pesticides and opting for the use of pesticides of biological origin will contribute to the preservation of people's health

A cathodic photoelectrochemical immunoassay with dual signal amplification for the ultrasensitive detection of DNA damage biomarkers

Toxicity screening and risk assessment of an overwhelmingly large and ever-increasing number of chemicals are vitally essential for ecological safety and human health. Genotoxicity is particularly important because of its association with mutagenicity, carcinogenicity and cancer. Phosphorylated histone H2AX (γH2AX) is an early sensitive genotoxic biomarker. It is therefore highly desirable to develop analytical methods for the detection of trace γH2AX to enable screening and assessment of genotoxicity. Here, we developed a novel cathodic photoelectrochemical (PEC) immunoassay with dual signal amplification for the rapid and ultrasensitive detection of γH2AX in cell lysates. A sandwich immuno-reaction targeting γH2AX was first carried out on a 96-well plate, using a secondary antibody/gold nanoparticle/glucose oxidase conjugate as the labeled detection antibody. The conjugate increased the production of H2O2 and thus provided the first mechanism of signal amplification. The immuno-reaction product containing H2O2 was then detected on a photocathode prepared from Bi2+xWO6 rich in oxygen vacancies, with H2O2 acting as electron acceptor. The oxygen vacancies acted as both adsorption and activation sites of H2O2 and thus enhanced the photocurrent, which provided another mechanism of signal amplification. As a result, an ultrasensitive immunoassay for γH2AX determination was established with a limit of detection of 6.87 pg/mL (S/N = 3) and a wide linear range from 0.01 to 500 ng/mL. The practicability of this assay was verified by detecting γH2AX in cell lysates exposed to known genotoxic chemicals. Our work offers a promising tool for the screening of genotoxic chemicals and opening a new avenue toward environmental risk assessment.

Hematological, Genotoxic and Oxidative Stress Response in Oreochromis niloticus (Nile Tilapia) Exposed to Varying Concentrations of Ethanolic Extract of Commelina benghalensis

This study evaluated the hematological, genotoxic and oxidative stress biomarkers of Oreochromisniloticus exposed to different concentrations of ethanolic extract of Commelina benghalensis(Dayflower). Sublethal exposure at 1% and 10% LC50 was done for twenty-one (21) days with physico-chemical parameters of cultured water and behavioral responses assessed. Hematological; genotoxic; antioxidant enzyme and non-enzyme activities studies were analyzed using standard methods. Results showed increase in the levels of Red blood cells; heamoglobin and white blood cells across extract concentrations compared to control. The number of micronucleated cells increased compared to control; though not significant difference (p&gt;0.05). Superoxide dismutase (SOD); reduced glutathione (GSH) and glutathionde-S-transferase (GST) activities were inhibited by the plant extract; while the level of lipid peroxidation as expressed by malondialdehyde (MDA) increased compared to control. Skin colour change; erratic swimming and mortality were observed at both higher concentration (10%) and lower concentration (1%) of the extract; though behavioral response at 10% extract concentration was higher than 1% extract concentration. In conclusion; the plant C.benghalensis has the potential to exert stress and promote cellular damage in contrast to its ability to boost the blood status of O. niloticus at varying concentration.

In vivo cytogenotoxicity testing of isotretinoin by the micronucleus assay in the blood of male Sprague-Dawley rats Isotretinoin is cytotoxic and genotoxic

Isotretinoin (ISO), one of vitamin A-derived retinoids, is comparatively the most widely prescribed drug in acne vulgaris pathogenesis. Despite its excellent therapeutic success, the systemic use of this drug showed undesirable serious side effects such as teratogenesis, oxidative stress, and genotoxicity. The uses of retinoids in cancer therapy are limited due to severe adverse reactions. This has led the scientific community to ask for further studies qualifying ISO features and comparing their efficacy and safety. The purpose of this study was to evaluate the cytogenotoxicity of multiple oral doses (5, 10, 15, and 20 mg/kg, daily for seven consecutive days) of ISO in male Sprague-Dawley rats. The micronucleus assay was used to investigate genotoxicity biomarkers such as the percentage of micronucleated polychromatic erythrocytes (%MNPCEs) and the percentage of aberrant cells (%Abc). Another goal was to test the cytotoxicity of the drug by measuring the ratio between PCEs and normochromatic erythrocytes (NCEs) (PCEs/NCEs). In comparison with the control, the three cytogenetic endpoints: %MNPCEs, P/N, and %Abc significantly (P ≤ 0.0017) dose-dependent increase. This suggested genotoxicity and cytotoxicity of the tested ISO doses. Therefore, the therapeutic uses of ISO should be restricted to a very narrow range border. Further studies are needed to shed more light on the safety profile of ISO therapy.

Associations between DNA methylation and genotoxicity among lead-exposed workers in China

Studies have shown that lead (Pb) exposure caused genotoxicity, however, the underlying mechanisms remain unclear. A mechanism may be via DNA methylation which is one of the most widely studied epigenetic regulations for cellular activities. Whether this is involved in Pb-induced genotoxicity has rarely been studied. Our study aimed to examine whether DNA methylation was associated with Pb exposure and genotoxicity, and to explore its potential mediating roles. A total of 250 Pb-exposed workers were enrolled. Blood lead levels (BLLs) and genotoxic biomarkers (Micronuclei and Comet) were analyzed. Methylation levels at CpG sites of LINE1 and Alu and promoter region of P53, BRCA1, TRIM36 and OGG1 were measured by pyrosequencing. Generalized linear model (GLM) combined with restricted cubic splines (RCS) were used to analyze relationships between Pb exposure, DNA methylation and genotoxicity. Mediation effect was used to explore mediating roles of DNA methylation. The distribution of BLLs was right-skewed and showed wide ranges from 23.7 to 636.2 μg/L with median (P25, P75) being 218.4 (106.1, 313.9) μg/L among all workers. Micronuclei frequencies showed Poisson distribution [1.94 ± 1.88‰] and Comet tail intensity showed normal distribution [1.69 ± 0.93%]. GLM combined with RCS showed that Alu methylation was negatively associated with BLLs, while P53 and OGG1 methylation were positively associated with BLLs. Micronuclei were negatively associated with Alu and TRIM36 methylation but positively with P53 methylation. Comet was positively associated with P53 and BRCA1 methylation. Mediation effect showed that Alu methylation mediated 7% effects on association between Pb exposure and micronuclei, whereas, P53 methylation mediated 14% and BRCA1 mediated 9% effects on association between Pb exposure and Comet. Our data show that Pb exposure induced changes of global and gene-specific DNA methylation which mediated Pb-induced genotoxicity.

Assessment of Protective Effects of Carvacrol on Haloperidol-Induced Oxidative Stress and Genotoxicity in Human Peripheral Blood Lymphocytes.

Haloperidol is a first-generation antipsychotic drug that has several indications in a wide range of mental conditions. The extensive prescription of haloperidol is correlated with some less-known adverse effects such as genotoxicity. Carvacrol is a monoterpenoid mainly found in oregano and thyme. It has the potential to scavenge free radicals in addition to increasing antioxidant defense enzyme activities and glutathione levels. In this study, we attempted to explore the possible potential of haloperidol in inducing genotoxicity in human peripheral lymphocytes as well as the protective role of carvacrol against this effect. The lymphocytes were divided into separate groups as follows: control group (cosolvent and NS); carvacrol group (5 μM); haloperidol group (25, 50, and 100 ng/ml); haloperidol (25, 50, and 100 ng/ml) + carvacrol (5 μM); positive control (0.8 μg/ml Cisplatin). After 24 hours of treatment, we conducted a cytokinesis-Block micronucleus test and an alkaline comet assay in order to determine genetic damage. Additionally, we measured glutathione and MDA levels as the biomarkers associated with oxidative stress. Significant increases in the levels of genotoxicity biomarkers (micronucleus frequency, DNA percentage in tail and tail moment) were observed in haloperidol-treated cells. The result of our oxidative stress tests also demonstrated that haloperidol had the potential to induce oxidative stress via reducing the levels of glutathione and increasing lipid peroxidation. Treatment with carvacrol significantly decreased the genotoxic events. It can be presumed that the induction of oxidative stress by haloperidol is the critical event associated with haloperidol-mediated genotoxicity. Therefore, using carvacrol as a natural antioxidant protected human lymphocytes against haloperidol genetic damage.

Environmental quality and ecotoxicity of sediments from the lower Salado River basin (Santa Fe, Argentina) on amphibian larvae

The lower Salado River basin receive agricultural, industrial and domestic waste water. So, the aim was to evaluate the quality of three sampling sites that belong to the Salado River basin (S1: Cululú stream; S2: Salado River, at Esperanza City, S3: Salado River at Santo Tomé City) based on physicochemical parameters, metals and pesticides analyses and ecotoxicity on Rhinella arenarum larvae. R. arenarum larvae (Gosner Stage -GS- 25) were chronically exposed (504h) to complex matrixes of surface water and sediment samples of each site for the determination of the survival rate. Biomarkers of oxidative stress, neurotoxicity and genotoxicity were analyzed in R. arenarum larvae (GS. 25) after exposure (96h) to the complex matrix of water and sediment. The water quality index showed a marginal quality for all sites, influenced mainly by low dissolved oxygen, high total suspended solid, phosphate, nitrite, conductivity, Pb, Cr and Cu levels. Metal concentrations were higher in sediment than in water samples (˜34-35000 times). In total, thirty different pesticides were detected in all water and sediment samples, S1 presented the greatest variety (26). Glyphosate and AMPA were detected in sediments from all sites, being higher in S3. N,N-Diethyl-meta-toluamide (DEET) and atrazine were detected in all water samples. Greatest mortality was observed in larvae exposed to samples from S1 from 288h (43.3%), reaching a maximum value of 50% at 408h. Oxidative stress and genotoxicity were observed in larvae exposed to S1 and S3 matrix samples. Neurotoxicity was observed in larvae exposed to all matrix samples. The integrated biomarker response index showed that larvae exposed to S1 and S3 were the most affected. According to the physicochemical data and the ecotoxicity assessment, this important river basin is significantly degraded and may represent a risk to aquatic biota, especially for R. arenarum larvae.

Bioaccumulation, genotoxicity, and risks to native fish species from inorganic contaminants in the Pantanal Sul-Mato-Grossense, Brazil

The Aquidauana River is one of the most important rivers in the Pantanal region, Brazil. However, its waters have been contaminated by nearby anthropogenic activities, threatening native fish species. In this study, our objectives were: 1) to determine the concentrations of Al, As, Cd, Cu, Fe, Mn, Ni, Pb, and Zn in water and sediment samples from the Aquidauana River and to assess the risks posed to aquatic biota; 2) to quantify the concentration of these elements in muscle and liver tissue samples from four native fish species; 3) to evaluate the potential bioaccumulation of inorganic elements in the muscles and liver; and 4) to investigate genotoxicity biomarkers and their association with the inorganic element concentrations present in the muscle tissue. Water and fish samples were collected in November 2020. The concentrations of Al, As, Cd, Cu, Fe, and Pb in the water samples were in disagreement with the Brazilian legislation and presented risks to the aquatic biota. In terms of mixtures of inorganic elements, there was a great increase in the risk to biota. The As concentration did not meet the Brazilian standard for sediments in the sample collected at sampling site 6. The concentrations of Cd and Pb in the muscle tissue of Hypostomus regani, Prochilodus lineatus, Brycon hilarii, and Mylossoma duriventre exceeded the Brazilian standards for human consumption. H. regani showed greater genotoxic damage, and the higher the Al and Fe concentrations in the muscle tissue, the higher the frequencies of lobulated nuclei and nuclear invaginations. Together, our results demonstrate the negative impacts on native fish species from the Aquidauana River contamination and indicate risks to Pantanal biodiversity.

Mercury bioaccumulation, genotoxic and biochemical biomarkers reveal the health status of yellow-spotted Amazon River turtles (Podocnemis unifilis) in an environmental protection area in the Amazon

ABSTRACT Chelonians are considered good bioindicators of environmental quality. The assessment of the health status of chelonian populations in the Amazon is important because they are traditionally consumed in large numbers in riverine communities and sustainable use reserves. The present study aimed to evaluate the health of Podocnemis unifilis (Testudines, Podocnemididae) in an environmental protection area in the Amazon region in Brazil. We analyzed the biomarkers lipoperoxidation, carbonylation of proteins, occurrence of micronuclei and erythrocytic nuclear abnormalities, quantified metallothioneins, and evaluated mercury bioaccumulation. We generated pioneering data regarding biomarkers in wild Amazonian freshwater turtles. All biomarker responses did not vary significantly between the sexes. The occurrence of oxidative and genotoxic damage, as well as concentrations of metallothioneins was low compared to other studies. In addition, the bioaccumulation of mercury in the muscle of the animals was below the limit recommended for human consumption by the World Health Organization. Our results provide baseline data on mercury bioaccumulation and biomarker responses that can be useful for future comparisons with other freshwater turtles. The data also provide evidence that the sustainable exploitation of these turtles in the study area (Piagaçu-Purus Sustainable Development Reserve) does not pose a risk to local riverine communities, as the detected mercury concentrations are safe for human consumption. In this sense, our results highlight the importance of protected areas for the conservation of healthy turtle populations, at the same time ensuring the health of the human populations that use them as a food resource.

HBM4EU Chromates Study-Genotoxicity and Oxidative Stress Biomarkers in Workers Exposed to Hexavalent Chromium.

A study was conducted within the European Human Biomonitoring Initiative (HBM4EU) to characterize occupational exposure to Cr(VI). Herein we present the results of biomarkers of genotoxicity and oxidative stress, including micronucleus analysis in lymphocytes and reticulocytes, the comet assay in whole blood, and malondialdehyde and 8-oxo-2′-deoxyguanosine in urine. Workers from several Cr(VI)-related industrial activities and controls from industrial (within company) and non-industrial (outwith company) environments were included. The significantly increased genotoxicity (p = 0.03 for MN in lymphocytes and reticulocytes; p < 0.001 for comet assay data) and oxidative stress levels (p = 0.007 and p < 0.001 for MDA and 8-OHdG levels in pre-shift urine samples, respectively) that were detected in the exposed workers over the outwith company controls suggest that Cr(VI) exposure might still represent a health risk, particularly, for chrome painters and electrolytic bath platers, despite the low Cr exposure. The within-company controls displayed DNA and chromosomal damage levels that were comparable to those of the exposed group, highlighting the relevance of considering all industry workers as potentially exposed. The use of effect biomarkers proved their capacity to detect the early biological effects from low Cr(VI) exposure, and to contribute to identifying subgroups that are at higher risk. Overall, this study reinforces the need for further re-evaluation of the occupational exposure limit and better application of protection measures. However, it also raised some additional questions and unexplained inconsistencies that need follow-up studies to be clarified.

Ecotoxicological assessment of complex environmental matrices from the lower Paraná River basin

Sediments of aquatic ecosystems constitute the fate of most atmospheric and terrestrial pollutants. Since aquatic organisms, such as amphibians, interact with sediments, the presence of pollutants may affect their survival, growth and reproduction. So, the aim of this study was to evaluate, the sediment and water quality of five sites from the lower basin of the Paraná River (Buenos Aires, Argentina) with different anthropic impacts: Morejón stream (S1), de la Cruz stream upstream (S2) and downstream (S3), Arrecifes river (S4), tributary stream of Arrecifes river (S5). Physicochemical parameters were measured in situ (water) and in laboratory (water and sediment samples). Also, a screening of metals and pesticides was performed. Chronic (504 h) lethal toxicity bioassays were performed exposing Rhinella arenarum larvae to sediment and water samples. Oxidative stress (catalase, superoxide dismutase, glutathione S transferase, reduced glutathione and lipid peroxidation) and genotoxicity (micronuclei test) biomarkers were analyzed at acute (96 h) exposure. According to the calculated water quality index, S1 and S3 showed excellent quality, S2 good quality and, S4 and S5 poor quality. Dissolved oxygen was low in all sites (2.26–5.63 mg/L) and S5 had the highest organic matter content. Copper levels exceeded the limit for the protection of aquatic life in S2 and S4; arsenic levels exceeded its limit in S4; and selenium levels exceeded its limit in S4 and S5. Pesticides were mainly detected in water samples. Sediment from S5 showed higher sulfide and organic matter concentrations. At 504 h, no significant mortality was observed in the control group while S5 caused the greatest mortality (80%), followed by S2 (66.67%), S1 (63.33%), S3 (46.67%) and S4 (43.4%). All samples caused oxidative stress and lipid peroxidation, and samples from S4 also caused genotoxicity. The analysis of sediment and water samples was a suitable approach to assess the effects of water bodies on a native amphibian species.

Automatic recognition of micronucleus by combining attention mechanism and AlexNet

BackgroundMicronucleus (MN) is an abnormal fragment in a human cell caused by disorders in the mechanism regulating chromosome segregation. It can be used as a biomarker for genotoxicity, tumor risk, and tumor malignancy. The in vitro micronucleus assay is a commonly used method to detect micronucleus. However, it is time-consuming and the visual scoring can be inconsistent.MethodsTo alleviate this issue, we proposed a computer-aided diagnosis method combining convolutional neural networks and visual attention for micronucleus recognition. The backbone of our model is AlexNet without any dense layers and it is pretrained on the ImageNet dataset. Two attention modules are applied to extract cell image features and generate attention maps highlighting the region of interest to improve the interpretability of the network. Given the problems in the data set, we leverage data augmentation and focal loss to alleviate the impact.ResultsExperiments show that the proposed network yields better performance with fewer parameters. The AP value, F1 value and AUC value reach 0.932, 0.811 and 0.995, respectively.ConclusionIn conclusion, the proposed network can effectively recognize micronucleus, and it can play an auxiliary role in clinical diagnosis by doctors.