The Incidence of the XRCC1 rs25487 and PON1 rs662 Polymorphisms in a Population from Central Brazil: Patterns in an Area with a High Level of Agricultural Activity.

In Brazil, high levels of agricultural activity are reflected in the consumption of enormous amounts of pesticides. The production of grain in Brazil has been estimated at 289.8 million tons in the 2022 harvest, an expansion of 14.7% compared with 2021. These advances are likely associated with a progressive increase in the occupational exposure of a population to pesticides. The Paraoxonase 1 gene (PON1) is involved in liver detoxification; the rs662 variant of this gene modifies the activity of the enzyme. The repair of pesticide-induced genetic damage depends on the protein produced by the X-Ray Repair Cross-Complementing Group 1 gene (XRCC). Its function is impaired due to an rs25487 variant. The present study describes the frequencies of the rs662 and rs25487 and their haplotypes in a sample population from Goiás, Brazil. It compares the frequencies with other populations worldwide to verify the variation in the distribution of these SNPs, with 494 unrelated individuals in the state of Goiás. The A allele of the rs25487 variant had a frequency of 26% in the Goiás population, and the modified rs662 G allele had a frequency of 42.8%. Four haplotypes were recorded for the rs25487 (G > A) and rs662 (A > G) markers, with a frequency of 11.9% being recorded for the A-G haplotype (both modified alleles), 30.8% for the G-G haplotype, 14.3% for the A-A haplotype, and 42.8% for the G-A haplotype (both wild-type alleles). We demonstrated the distribution of important SNPs associated with pesticide exposure in an area with a high agricultural activity level, Central Brazil.

Current Status of Ecotoxicological Studies of Bats in Brazil.

Bats are sensitive to contaminants generated by agricultural activities, mining, and urbanization. In this review, we update the status of bat toxicology in Brazil. Agriculture, for example, in addition to habitat fragmentation and loss, undoubtedly affects non-target organisms through the use of pesticides. Other factors such as trace metals are a neglected problem in the country, as they can deposit on insects and plants reaching bats through the ingestion of these foods. Of the 184 species of bats in the country, only 4.9% have been investigated. The frugivorous species, Artibeus lituratus, has frequently been studied for the effects of pesticide exposure, and impacts at the cellular level on metabolism and reproduction have been observed. Given the scarcity of studies on bat ecotoxicology, we encourage national researchers and scientists elsewhere to increase knowledge of the effects of chemical contaminants on bats in Brazil.

Assessing Genotoxicity and Mutagenicity of Three Common Amphibian Species Inhabiting Agroecosystem Environment.

Amphibians are constantly exposed to pollutants and the stress of agricultural activities. We selected three anuran amphibian species Dendropsophus minutus, Boana albopunctata, and Physalaemus cuvieri, totaling 309 individuals. We collected tadpoles in 15 permanent ponds: 5 soybean crops, 3 corn crops, and 7 nonagricultural lands. Our study provides the first comparative data on the genotoxicity and mutagenicity of three common amphibian anurans. Dendropsophus minutus was the most vulnerable species compared with B. albopunctata and P. cuvieri for comet assay and micronuclei test. However, the more significant amount of DNA damage seen in D. minutus does not mean that their populations are threatened once such species adapt well to anthropogenic disturbances. Despite, P. cuvieri was less sensitive than the other two species; the DNA damage was significantly higher in soybean crops. Physalaemus cuvieri is a leptodactylidae species that deposit their eggs in foam nests, which are essential to protect eggs from dehydration. Moreover, the foam reduces the contact of eggs with water; thus, P. cuvieri eggs could be less exposed to contaminants present in pounds, compared with D. minutus and B. albopunctata, which deposit their eggs directly in the water. Therefore, this study was sufficiently sensitive to detect genotoxic and mutagenic effects in tadpoles exposed to agroecosystems. We strongly suggest D. minutus in future biomonitoring studies that involve the comparison of anthropized versus not anthropized environments. Overall, we recommend the comet assay and micronucleus test as effective methods for the detection of genotoxic damage in amphibian anurans to the environmental disturbance, especially in agricultural sites.

Chemical, ecotoxicological, cytotoxic, and mutagenic evaluation of gelling agents used in the production of 70% alcohol gel.

With COVID-19, there has been an increase in the use of gelling agents for hand sanitizer production, and as a result, the release of this product into wastewater could induce impacts and adverse reactions in living organisms. Thus, ecotoxicological and cytotoxicological assessments of gelling agents with test organisms from different trophic levels are necessary to assess their environmental safety. For this, seven cellulose-based gelling agents and a polyacrylic acid derivative (C940) were selected for tests with Artemia salina. The most toxic agent was tested on Allium cepa to assess cytotoxicity. The volatile compounds of the gelling agents were analyzed. Cellulose-based gelling agents were not considered toxic according to their LC50, but C940 presented moderate toxicity to A. salina and cytotoxicity to Allium cepa, but without mutagenicity. In addition, C940 contained cyclohexane as a volatile compound. Thus, cellulose-based gelling agents are better environmental options than carbomer for 70% alcohol gel sanitizer.

Influence of different concentrations of plasticizer diethyl phthalate (DEP) on toxicity of Lactuca sativa seeds, Artemia salina and Zebrafish.

Like other phthalates, diethyl phthalate (DEP) is considered as a contaminant of emerging concern (CEC) due to its ease in migrating from a package to water and food, and hence contaminate consumers, being metabolized and excreted in the urine. Its presence has a negative impact on aquatic ecosystems, especially with respect to disruption of the endocrine system and to reproductive disorders in humans. It mainly enters water bodies via sewage effluents from effluent treatment plants, due to its incomplete or inefficient removal. The objective of this work was to evaluate the toxicity of DEP at different trophic levels and to analyze data on the incidence and concentration of DEP according to its solubility. The concentrations ranged from 12.5 mg L-1 to 500 mg L-1 considering the response for toxicity at each trophic level and to determine the lethal concentration in 50% of the following organisms (LC50) (in mg L-1): Lactuca sativa seeds, Artemia salina Leach nauplii and Zebrafish embryo larval stage (Danio rerio), being 41,057.58 after 120 h; 401.77 after 48 h; and 470 after 96 h of exposure, respectively. As expected, higher organisms were more affected even at low concentrations, which shows the anthropological contribution of CECs to water bodies.

The effect of low-dose exposure on germline microsatellite mutation rates in humans accidentally exposed to caesium-137 in Goiânia.

A serious radiological accident occurred in 1987 in Goiânia, Brazil, which lead to extensive human and environmental contamination as a result of ionising radiation (IR) from caesium-137. Among the exposed were those in direct contact with caesium-137, their relatives, neighbours, liquidators and health personnel involved in the handling of the radioactive material and the clean-up of the radioactive sites. The exposed group consisted of 10 two-generation families, totalling 34 people. For each exposed family, at least one of the progenitors was directly exposed to very low doses of γ-IR. The control group consisted of 215 non-irradiated families, composed of a father, mother and child, all of them from Goiânia, Brazil. Genomic DNA was purified using 100 µl of whole blood. The amplification reactions were prepared according to PowerPlex® 16, following the manufacturer's instructions. Genetic profiles were obtained from a single polymerase chain reaction amplification. The exposed group had only one germline mutation of a paternal origin in the 'locus' D8S1179 and the observed mutation presented a gain of only one repeat unit. In the control group, 11 mutations were observed and the mutational events were distributed in five loci D16S539, D3S1358, FGA, Penta E and D21S11. The mutation rates for the exposed and control groups were 0.006 and 0.002, respectively. There was no statistically significant difference (P = 0.09) between the mutation rate of the exposed and control groups. In conclusion, the quantification of mutational events in short tandem repeats can provide a useful system for detecting induced mutations in a relatively small population.

Evaluation of the genotoxic, mutagenic, and histopathological hepatic effects of polyoxyethylene amine (POEA) and glyphosate on Dendropsophus minutus tadpoles.

Herbicides improve the productivity of a monoculture by eliminating weeds, although they may also be toxic and have negative effects on non-target organisms, such as amphibians. The present study evaluated the genotoxic, mutagenic, and histopathological hepatic responses of Dendropsophus minutus tadpoles to acute exposure (96 h) to the herbicide glyphosate (GLY, 65, 130, 260 and 520 µg/L) and the surfactant polyoxyethylene amine (POEA, 1.25, 2.5, 5 and 10 µg/L). On average, 174 % more genomic damage was observed in the tadpoles exposed to all concentrations of POEA in comparison with the control, while up to seven times more micronuclei were recorded, on average, at a concentration of 5 µg/L of POEA. All the individuals exposed to 10 µg/L of POEA died. The tadpoles exposed to GLY presented 165 % more DNA damage than the control, on average, at the highest concentrations (260 and 520 µg/L), and up to six times more micronuclei at 520 µg/L. The Erythrocyte Nuclear Abnormality test (ENA) detected a relatively high frequency of cells with lobed nuclei in the tadpoles expose to POEA at 5 µg/L and binucleated cells in those exposed to GLY at 520 µg/L. The hepatic histopathological observations revealed several types of lesions in the tadpoles exposed to both GLY and POEA. Overall, then, the results of the study indicate that both GLY and POEA have potential genotoxic, mutagenic, and hepatotoxic effects in D. minutus tadpoles. We emphasize the need for further studies to monitor the amphibian populations, such as those of D. minutus, which breed in aquatic environments associated with agricultural areas. The release of pollutants into natural habitats may have significant long-term impacts on the survival of anuran tadpoles.

Cultivated areas and rural workers' behavior are responsible for the increase in agricultural intoxications in Brazil? Are these factors associated?

In Brazil, the increased use of pesticides has caused several consequences such as mortality of non-target species, contamination of aquatic environments, and changes in biodiversity. Regarding humans, such products could increase the cases of poisoning, the number of different types of cancer, and also neurodegenerative and autoimmune diseases, especially in workers who occupationally handle pesticides. Therefore, this study aimed to evaluate the number of agricultural poisonings from 2007 to 2017 and if there is a correlation between harvested areas, and to analyze the possible factors that contribute to poisonings. Our results demonstrated an increase in poisonings by 8% and a correlation with the growth of the harvested area. We emphasize that other factors are also determinant for the increase of poisonings such as the absence or inappropriate use of personal protective equipment (PPE), the toxicity of mixtures, the mishandling, and the lack of information on how to use and spray such products. Among the circumstances about poisoning, we presented suicide attempts and accidental causes. We found that the area planted and harvested is also associated with the amount of poisoning; however, several other behavioral factors are also determinant for the growth of agricultural poisoning in Brazil.

The importance of understanding the distribution of GSTM1 and GSTT1 genotypes and haplotypes in a region with intense agriculture activity.

Brazil is one of the largest pesticide consumers in the world, mainly due to its intense agricultural activity. The State of Goias, situated in Central Brazil, is a region recognized as an essential producer of soy, corn, beans, sorghum, sugar cane, and cotton. In this study, we evaluated 602 unrelated individuals, distributed in central and southern regions in Goias, presenting combined frequencies (haplotypes) of the GSTT1 and GSTM1 genes. In all municipalities, the frequency of the GSTT1 null genotype was 38.2% and of the GSTM1 null genotype was 50.3%. Goiania, the capital of Goias, presented the highest frequencies of GSTT1 and GSTM1 null genotypes, probably due to a founder effect of non-representative colonizing ancestors. So, the ancestral population adapted to the environment, with the frequencies observed in Goiania. However, nowadays, as there is excessive use of pesticides, the community becomes susceptible to the harmful effects of xenobiotics exposure, mainly due to the high frequency of GSTT1 and GSTM1 null genotypes. As in Goias, the consumption of pesticides has shown considerable growth, haplotypes with null alleles are of high risk for the population. Our results indicated that it is essential to understand the frequencies of the GSTT1 and GSTM1 genes for the monitoring of risk groups, like farmers, who have contact with pesticides, directly or indirectly, as well as assisting in the development of preventive medicine practices.

Increased DNA damage is not associated to polymorphisms in OGGI DNA repair gene, CYP2E1 detoxification gene, and biochemical and hematological findings in soybeans farmers from Central Brazil.

Our study evaluated 163 individuals, being 74 soybean farmers, occupationally exposed to pesticides, and 89 individuals from Goias municipalities, Central Brazil, with similar conditions to the exposed group, comprising the control group. Of the 74 soybean farmers, 43 exposed directly to pesticides and 31 exposed indirectly. The exposed group consisted of individuals aged 19 to 63 years, 21 women and 53 men, and the control group had ages ranging from 18 to 64 years, being 36 women and 53 men. 18.9% of the exposed group were poisoned by pesticides, and the most common symptoms were headache and gastrointestinal problems. The genotype frequencies of the rs2031920 (T>C) polymorphism in the CYP2E1 gene present significant differences between the exposed and control groups (p = 0.02), showing that 24.3% of the exposed group were heterozygotes against 6.7% in the control group. For the OGG1 gene, two SNPs, rs1052133 (G>C) and rs293795 (T>C), were evaluated and the genotype frequencies were not statistically different between the exposed and control groups. The DNA damage was distinct (p < 0.05) in the three analyzed comet parameters (tail length, Olive tail moment, %DNA) between groups. However, there was no influence of age and alcohol consumption between the groups associated with the polymorphisms in the CYP2E1 and OGG1 genes and DNA damage. We also did not find altered hematological and biochemical parameters in the exposed group. Thus, this pioneering study at Goias State carried out an overview of the health of soybean farmers. We evaluated classic laboratory exams, associated with exposure markers (comet assay) and susceptibility markers (genetic polymorphisms), emphasizing the need to expand the Brazilian health assessment protocol. We found, in soybean farmers, increased DNA damage and a higher number of heterozygotes in CYP2E1 gene, compared with the control group, despite the lack of association with age, educational level, smoking, drinking habits, and genetic polymorphisms.

Evaluating the OGG1 rs1052133 and rs293795 polymorphisms in a sample of rural workers from Central Brazil population: a comparative approach with the 1000 Genomes Project.

Brazilian population is one of the largest consumers of pesticides in the world, especially the Central Brazil population. Thus, the aim of this study was to evaluate the frequency of genotypes, alleles, haplotypes, and the linkage disequilibrium (DL) of the OGG1 gene in rural workers from Central Brazil, comparing with the populations of the 1000 genome. Three hundred thirty healthy individuals not related and randomly selected were included in this study. We obtained genomic DNA from peripheral blood lymphocytes. The 748-bp OGG1 gene was amplified by PCR and sequenced. Of the 330 individuals, 215 (65%) were males and 115 (35%) were females. There were no differences in the distribution of the rs1052133 and rs293795 with age and sexes. Haplotypes containing only conserved T/C alleles were the most common in our population. The frequency of the mutant alleles of rs1052133 and rs293795, in our population, was 20% and 30%, respectively, and it is noteworthy, worldwide, that mutant alleles are commonly associated to an increased risk for the development of cancer, specially due to direct or indirect contact to pesticides, as occurs in rural workers of Central Brazil population.

Investigation of CAV1/CAV2 rs4236601 and CDKN2B-AS1 rs2157719 in primary open-angle glaucoma patients from Brazil.

Large-scale genome-wide association studies have identified several susceptibility variants associated with the risk of primary open-angle glaucoma (POAG), among which rs4236601 (CAV1/CAV2) at chromosome 7q31 and rs2157719 at chromosome 9p21 (CDKN2B-AS1). The purpose of this study was to investigate whether these variants contribute to the incidence of POAG in a sample of the Brazilian Southeastern population and to determine the best-fitted genetic model for these single nucleotide polymorphisms (SNPs). A case-control study with 557 individuals, 310 with POAG, and 247 controls was conducted through PCR and direct sequencing. We observed a significant effect of the heterozygous genotype (G/A) of rs2157719 that occurred more frequently in the control group (p = 0.0004; OR: 0.517, CI 95%: 0.357-0.745). Allele frequencies also differed between cases and controls (p = 0.006; OR: 0.694, CI 95%: 0.522-0.922) with the best-fitted genetic model for rs2157719 being the codominant model. No differences were observed for genotype and allele distributions in relation to rs4236601 in the CAV1/CAV2 region. The association of rs2157719 (CDKN2B-AS1) with the POAG phenotype corroborates previously published results, reinforcing the importance of this variant in POAG etiology.