Brominated flame retardants (BFRs) are ubiquitous and persistent environmental contaminants owing to their extensive use in consumer products. Although linked to various adverse health effects, the underlying molecular mechanisms remain complex. This review consolidates scientific evidence positioning mitochondria as a central target of BFR toxicity, unraveling the pathways that drive cellular damage. The analysis revealed that BFRs converge on the fundamental mechanisms of mitochondrial injury. They consistently impair bioenergetics by disrupting the electron transport chain and uncoupling oxidative phosphorylation, leading to ATP depletion and collapse of the mitochondrial membrane potential (ΔΨm). This energetic failure triggers a surge in reactive oxygen species, overwhelming antioxidant defenses, and causing severe oxidative damage. Beyond these common effects, this review highlights remarkable mechanistic plasticity. Tetrabromobisphenol A can induce distinct cell death programs, including apoptosis, necroptosis, and ferroptosis, depending on the cellular context of the study. Furthermore, BFR biotransformation can yield metabolites such as hydroxylated polybrominated diphenyl ethers (PBDEs) that exhibit significantly greater toxicity than their parent compounds. Finally, mitochondrial dysfunction is a central hub that orchestrates cellular damage by BFRs. This is critically highlighted by the replacement of BDE-209 with decabromodiphenyl ethane, a regrettable substitution, where the new compound shares similar mitotoxic mechanisms and has become a widespread pollutant. This underscores the urgent need for a paradigm shift toward mechanism-based risk assessment to prevent future cycles of hazardous chemical replacements and to guide the design of genuinely safer alternatives.

Fipronil sulfone is a persistent oxidative metabolite of the insecticide fipronil, frequently detected in aquatic systems and known to exert neurotoxic effects on non-target organisms. Despite its environmental relevance, the developmental impacts of fipronil sulfone remain underexplored. In this study, zebrafish embryos and larvae were exposed to environmentally relevant concentrations of fipronil sulfone (12.5-1000 μg/L) at different developmental stages (24, 48, 72, 120, and 144 hpf). Morphological, behavioral, and biochemical endpoints were assessed. Exposure to concentrations ≥125 μg/L significantly reduced swim bladder inflation (up to 100 % inhibition at ≥250 μg/L) and induced spinal curvature in over 60 % of larvae. Survival decreased progressively with increasing concentrations, reaching 100 % mortality at 1000 μg/L. Larvae exposed to 125 μg/L also exhibited hyperactivity followed by reduced locomotion, likely due to blocked GABAergic receptors and acetylcholinesterase inhibition. Moreover, oxidative stress was confirmed by increased reactive oxygen species and glutathione S-transferase activity. These findings demonstrate that even low concentrations of fipronil sulfone can compromise neurodevelopment and survival in zebrafish, underscoring the need for its inclusion in environmental monitoring and risk assessment frameworks.

Diisopentyl phthalate (DiPP) is present in many consumer goods, but can be absorbed into the human body, and can disrupt the endocrine system affecting reproductive health and fetal development. Studies revealed that biological samples of pregnant women in Brazil contained DiPP, raising even more the concerns about its usage. This study investigated how DiPP concentrations (12.5-1000 mu g l-1) affect the different developmental stages (24, 48, 72, 120, and 144 hpf) of embryos and larvae of zebrafish. DiPP induced deleterious alterations in neuromuscular development and morphometry of organs, Low concentrations of DiPP decreased acetylcholinesterase and cellular energy allocation concomitantly with increased glutathione S-transferase. Zebrafish swimming period seemed to be decreased not only due to direct neurotoxicity, but also to less allocation of energy for behavioral purposes. Moreover, high concentrations of DiPP induced spinal deformities and developmental alterations specially of the eye and liver of larvae. These findings emphasize that DiPP exerts complex effects that should be considered when assessing its potential effects on health of humans and the total environment. The biomarkers and behavioral parameters showed to be good complimentary early-warning tools exhibiting a high sensitivity compared to FET.

Mitochondria are affected by chemical substances and play a critical role in drug-induced liver injury (DILI). Chemical substances can have a significant impact on various cellular processes, such as the disruption of oxidative phosphorylation, oxidative stress, and alteration of glucose metabolism. Given the consequences of these effects, it is crucial to understand the molecular pathways of chemical substances in the context of hepatotoxicity to prevent and treat DILI. In this regard, the Seahorse XFe24 Analyzer is a valuable tool for assessing mitochondrial bioenergetics and glucose metabolism. The Mito Stress Test and Glycolytic Rate Assay allow real-time assessment of the metabolic state after chemical exposure. Additionally, HepG2 spheroids have emerged as an important alternative tool for assessing hepatotoxicity, as they provide results that are more comparable to those found in humans than monolayer cultures or animal tests (such as rodent tests). By integrating these two powerful tools, it is possible to bridge the gap between animal and human tests, resulting in more reliable results in the assessment of human hepatotoxicity and DILI. However, because of the high variability in characteristics between 3D cultures (such as spheroids and organoids), XF analyzer assays are not well optimized for use with HepG2 spheroids. Here, we describe a streamlined and optimized protocol for performing the Mito Stress Test and Glycolytic Rate Assay using HepG2 spheroids cultured in alginate microcapsules in the Seahorse XFe24 Analyzer.

Flame retardants, crucial for fire prevention, are used worldwide, but they are considered to be ‘emerging contaminants’ and may pose risks to human and environmental health. Aluminum diethyl phosphinate (ALPI) is a halogen-free flame retardant. To evaluate the toxicity of this compound, the following assays were performed: Salmonella/microsome mutagenicity assay; toxicity assays with two endpoints (mitochondrial dehydrogenase activity, plasma membrane integrity); micronucleus assay with human hepatoma cell line HepG2. ALPI was not mutagenic in Salmonella strains TA97, TA98, TA100, TA102, or TA104. ALPI was not cytotoxic at any concentration tested. The HepG2 micronucleus assay showed genotoxicity of ALPI at 200 µg/mL and no cytotoxicity (cytokinesis-block proliferation index, CBPI). Our data are relevant to the regulation of flame retardants.

Fungicides are often used prophylactically, to control fungal diseases. Although fungicides have been designed to control pests/fungi, they frequently share molecular targets with non-target species, including humans. Tebuconazole, a fungicide belonging to the class of triazoles, is widely employed, has moderate to high persistence in soil, and can be found in different environmental levels. This fungicide is metabolized to the main hydroxy-derived metabolite, Tebuconazole-tert-butyl-hydroxy (or hydroxytebuconazole). This study aims to unveil the action mechanism of Tebuconazole and the role played by its metabolite, Tebuconazole-tert-butyl-hydroxy (5-(4-Chlorophenyl)-2,2-dimethyl-3-(1H-1,2,4-triazol-1-ylmethyl)-1,3-pentanediol), within the expected spectrum of toxicity. In silico and in vitro analyses (MTT assay, cell cycle evaluation, annexin/PI assay, ROS accumulation assay, and mitochondrial membrane potential determination) were performed in HepG2 cells for 24 h and 48 h. Although in silico analysis suggested that both Tebuconazole and Tebuconazole-tert-butyl-hydroxy are potentially hepatotoxic, only Tebuconazole affected the tested cell line. Reduced MTT metabolism, and decreased mitochondrial membrane potential were the main findings. In conclusion, the action mechanism of Tebuconazole may be related to mitochondrial dysfunction. However, the findings of this study pointed out that Tebuconazole-tert-butyl-hydroxy does not play an important role in Tebuconazol toxicity. The study has generated new data that will help to understand how fungicides behave in the environment.

Some dyes currently used by the textile, pharmaceutical, food, cosmetic, and photographic industries have been shown to be toxic and/or mutagenic to aquatic life. Most of these dyes resist degradation processes available for treating wastewater, and these processes might generate even more toxic by-products. Despite the large number of available dyes and the large quantity of dyes released into the environment, studies on their toxicity are still scarce. We evaluated and compared the effects in the animal model Danio rerio (zebrafish) of environmentally relevant concentrations of Disperse Red 60 (DR 60), 73 (DR 73), and 78 (DR 78) using the fish embryo acute toxicity (FET) test, morphometric analysis, immunofluorescence imaging, and behavioral parameters. DR 60 caused ocular modifications, while the DR 73 caused non-inflation of the swim bladder (NISB), pericardial edema (PE), scoliosis (S) and abnormal yolk sac (AYS) from at 0.125 mg/L. In behavioral tests, all the dyes induced changes in velocity and time spent swimming of exposed larvae. However, these alterations in behavior seem to be caused by different factors dependent on the dye and its concentration. Nevertheless, behavior seems to add valuable information concerning the hazards analysis of dyes, since it reveals to be the most sensitive group of parameters tested in the current study. In conclusion, of the behavioral and developmental alterations caused by these dyes should be interpreted as an alert for greater attention when registering new dyes and releasing them into the environment. In the particular case of DR 60 the possibility that directly affects the eye of larvae is of great environmental concern, but also from the human health perspective.

Fungicides are specifically used for controlling fungal infections. Strobilurins, a class of fungicides originating from the mushroom Strobilurus tenacellus, act on the fungal mitochondrial respiratory chain, interrupting the ATP cycle and causing oxidative stress. Although strobilurins are little soluble in water, they have been detected in water samples (such as rainwater and drinking water), indoor dust, and sediments, and they can bioaccumulate in aquatic organisms. Strobilurins are usually absorbed orally and are mainly eliminated via the bile/fecal route and urine, but information about their metabolites is lacking. Strobilurins have low mammalian toxicity; however, they exert severe toxic effects on aquatic organisms. Mitochondrial dysfunction and oxidative stress are the main mechanisms related to the genotoxic damage elicited by toxic compounds, such as strobilurins. These mechanisms alter genes and cause other dysfunctions, including hormonal, cardiac, neurological, and immunological impairment. Despite limitations, we have been able to compile literature information about strobilurins. Many studies have dealt with their toxic effects, but further investigations are needed to clarify their cellular and underlying mechanisms, which will help to find ways to minimize the harmful effects of these compounds.

The search for new methods in the toxicology field has increased the use of early life stages of zebrafish (Danio rerio) as a versatile organism model. Here, we use early stages of zebrafish to evaluate glyphosate as pure active ingredient and within a commercial formulation in terms of oxidative stress. Biomarkers involved in the oxidative status were evaluated along with other markers of neurotoxicity, genotoxicity, cytotoxicity, energy balance and motor performance, and the selected tools were evaluated by its sensitivity in determining early-warning events. Zebrafish embryos exposed to glyphosate active ingredient and glyphosate-based formulation were under oxidative stress, but only the commercial formulation delayed the embryogenesis, affected the cholinergic neurotransmission and induced DNA damage. Both altered the motor performance of larvae at very low concentrations, becoming larvae hypoactive. The energy balance was also impaired, as embryos under oxidative stress had lower lipids reserves. Although data suggest that glyphosate-based formulation has higher toxicity than the active ingredient itself, the most sensitive biomarkers detected early-warning effects at very low concentrations of the active ingredient. Biochemical biomarkers of defense system and oxidative damage were the most sensitive tools, detecting pro-oxidant responses at very low concentrations, along with markers of motor performance that showed high sensitivity and high throughput, suitable for detecting early effects linked to neurotoxicity. Alterations on morphology during embryogenesis showed the lowest sensitivity, thus morphological alterations appeared after several alterations at biochemical levels. Tools evaluating DNA damage and cell proliferation showed mid-sensitivity, but low throughput, thus they could be used as complementary markers.

Understanding the interplay among organophosphorus flame retardants (OPFRs), microplastics, and freshwater organisms is crucial for unravelling the dynamics within freshwater environments and foreseeing the potential impacts of organic pollutants and plastic contamination. For that purpose, the present research assessed the exposure impact of 10 mg L−1 flame-retardant aluminium diethylphosphinate (ALPI), 10 μg mg−1liver microplastics polyurethane (PU), and the combination of ALPI and PU on the freshwater planarian Girardia tigrina. The exposure to both ALPI and PU revealed a sequential effect, i.e., a decrease in locomotor activity, while oxidative stress biomarkers (total glutathione, catalase, glutathione S-transferase, lipid peroxidation) and metabolic responses (cholinesterase activity, electron transport system, and lactate dehydrogenase) remained unaffected. Despite this fact, it was possible to observe that the range of physiological responses in exposed organisms varied, in particular in the cases of the electron transport system, cholinesterase activity, glutathione S-transferase, catalase, and levels of total glutathione and proteins, showing that the energetic costs for detoxification and antioxidant capacity might be causing a lesser amount of energy allocated for the planarian activity. By examining the physiological, behavioural, and ecological responses of planarians to these pollutants, insights can be gained into broader ecosystem-level effects and inform strategies for mitigating environmental risks associated with OPFRs and microplastic pollution in freshwater environments.

MDMA (3,4-methylenedioxymethamphetamine), an entactogen with empathogenic and prosocial effects, is widely used in music festivals and other festive settings. High MDMA doses have been associated with drug-induced liver injury and cases of hyperthermia. Although the latter condition is thought to increase MDMA hepatotoxicity, this correlation remains poorly explored for recreational MDMA doses. On the other hand, the fact that MDMA acts to extinguish fear and to reconsolidate memory could be explored as an adjunct to psychotherapy during treatment of neuropsychiatric disorders such as post-traumatic stress disorder. In this context, assessing MDMA toxicity is relevant, and tridimensional cell culture has emerged as an alternative to animal models in toxicity assessment. Herein, we have used HepG2 spheroids to evaluate MDMA-induced hepatotoxicity at recreational doses, under normo- or hyperthermia. The MTT reduction assay did not evidence significantly reduced cell viability. Moreover, MDMA did not increase reactive oxygen species production, deplete the mitochondrial membrane potential, arrest the cell cycle, or induce apoptotic cell death. These findings support further pre-clinical investigation of MDMA safety from the perspective of both harm reduction and therapy given that non-abusive recreational and therapeutic doses overlap.

Produced water (PW) generated by oil companies is a highly impacting waste that contains chemicals such as metals and organic and inorganic compounds. Given its polluting potential, PW requires effective treatment before being discharged into the environment. Conventional treatments have limited efficiency in removing PW toxicity, so alternative approaches must be developed and standardized. In this context, treatment with adsorbent materials like magnetized vermiculite (VMT-mag) is highlighted. This work aimed to evaluate the efficiency of treatment with VMT-mag in reducing PW toxicity to aquatic biota. For this purpose, three aquatic species (the midge Chironomus riparius, the planarian Girardia tigrina, and the crustacean Daphnia magna) were exposed to untreated PW and to PW treated with VMT-mag at laboratory conditions. The assessed endpoints included mortality, growth, emergence, and developmental time of C. riparius; mortality, locomotion, feeding, and head regeneration of G. tigrina; and intrinsic population growth rate (r) and reproductive output of D. magna. The results showed that all the species exposed to raw PW were impaired: C. riparius had delayed development, G. tigrina had reduced locomotor activity and delayed head regeneration, and D. magna had reduced reproduction and delayed intrinsic population growth rate (r). Most of the analyzed parameters showed that treatment with VMT-mag diminished PW toxicity. Therefore, using VMT-mag to treat PW may be the key to reducing the PW effects on aquatic organisms.

Department of Chemistry, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, São Paulo, Brazil, Department of Clinical Analyses, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, São Paulo, Brazil, Department of Pharmacology, School of Medicine of Ribeirão Preto, University of São Paulo, São Paulo, Brazil, Department of Molecular Pharmacology, Albert Einstein College of Medicine, New York City, NY, United States

Polybrominated Diphenyl Ethers (PBDEs) are a major class of brominated flame retardants, and their widespread use has led them to be considered contaminants with emerging concern. PBDEs have been detected in the indoor air, house dust, food, and all environmental compartments. The congener BDE-47 (2,2 ',4,4 '-tetrabromodiphenyl ether) is the most prevalent, and hepatotoxicity, neurotoxicity, immunological changes, endocrine disruption, and genotoxic potential have been related to its exposure. Although the BDE-47 molecular toxicity pathway is directly related to intrinsic apoptotic cell death, the role of autophagy in BDE-47 toxicity remains unclear. In this context, three-dimensional cell culture has emerged as a good strategy for the replacement of animals in toxi-cological testing. Here, we used HepaRG spheroids cultured in alginate microcapsules to investigate the role of autophagy in BDE-47-mediated hepatotoxicity. We developed mature and functional HepaRG spheroids by culturing them in alginate microcapsules. Histological analysis revealed that HepaRG spheroids formed an extracellular matrix and stored glycogen. No apoptotic and/or necrotic cores were observed. BDE-47 showed concentration-and time-dependent cytotoxicity in HepaRG spheroids. In the early exposure period, BDE-47 initially disrupted mitochondrial activity and increased the formation of acid compartments that promoted the increase in autophagic activity; however, this autophagy was blocked, and long-term exposure to BDE-47 promoted efficient apoptotic cell death through autophagy blockade, as evidenced by an increased number of fragmented/condensed nuclei. Therefore, for the first time, we demonstrated BDE-47 toxicity and its cell pathway induces cell death using a three-dimensional liver cell culture, the HepaRG cell line.

Benzotriazoles are heterocyclic compounds typically presenting a benzene ring fused with a triazole molecule. The industry uses these compounds as anti-corrosion agents and recently, they have been employed in the pharmaceutical industry and in detergent formulations. Benzotriazoles persist in the environment, and water treatment plants cannot degrade them completely. Consequently, these compounds have been detected in rivers, lakes, and drinking water, which makes assessing their safety for the human and aquatic animal populations crucial. Here, we have evaluated and compared how exposure to 1H-benzotriazole or 5-chloro-benzotriazole affect the zebrafish embryo-larval stages. We have determined the acute toxicity, morphometric alterations, and acetylcholinesterase activity on zebrafish embryos, as well as behavioral endpoints using the tail coiling assay. The estimated LC50 of 5-chloro-benzotriazole was 19 mg/L, whereas 1H-benzotriazole caused no mortality. The zebrafish embryos exposed to 20 and 25 mg/L 5-chloro-benzotriazole had decreased hatching rate and exhibited pericardial and yolk sac edemas. Furthermore, the embryo length and eye area were decreased, in contrast with an increased yolk sac after exposure to 20 mg/L 5-chloro-benzotriazole. In turn, 1H-benzotriazole also decreased the eye area of zebrafish embryos, but no other significant morphological alterations were observed. The tail coiling assay showed that the zebrafish embryos increased the percentage of time moving and the number of embryonic movements per minute after exposure to 1H-benzotriazole (15 mg/L) or 5-chloro-benzotriazole (20 and 25 mg/L), indicating that these compounds were potentially neurotoxic. However, acetylcholinesterase activity was not significantly altered in embryos exposed to 1H-benzotriazole, but significantly decreased when exposed to 0.05 mg/L 5-chloro benzotriazole confirming its neurotoxicity at a much lower concentration. Our findings showed that 5-chloro-benzotriazole seems to induce more harmful alterations to zebrafish embryos than 1H-benzotriazole. Nevertheless, 1H-benzotriazole seems to induce a direct effect on eye development for concentrations lower than the ones of 5-chloro-benzotriazole affecting zebrafish embryos.

Tebuthiuron (TBU), a phenylurea herbicide, is widely applied in agricultural and non-agricultural soils. Because TBU resists degradation, it can contaminate water and reach the biota once it is released into the environment. However, the potential toxic effects of TBU on aquatic developing organisms have been poorly studied. By taking advantage of the early-life stages of zebrafish (Danio rerio), we have combined morphological, biochemical, behavioural, and molecular approaches to investigate the developmental toxicity triggered by environmentally relevant concentrations (from 0.1 to 1000 μg/L) of TBU. Exposure to TBU did not elicit morphological abnormalities but it significantly delayed hatching. In addition, TBU altered the frequency of tail coils in one-day post-fertilization (dpf) old embryos. Moreover, TBU exposure during four days significantly inhibited the whole body AChE activity of larvae. At the molecular level, TBU did not significantly affect the mRNA levels of four genes (elavl3, gfap, gap43, and shha) that play key roles during the neurodevelopment of zebrafish. By assessing the motor responses to repeated light-dark stimuli, 6 dpf larvae exposed to TBU displayed hyperactivity, showing greater travelling distance during the dark periods. Our categorization of swimming speed revealed an interesting finding – after the light was turned off, the exposed larvae abandoned the freezing mode (<2 mm/s) and travelled mainly at cruising speed (2–20 mm/s), showing that the larval hyperactivity did not translate into higher swimming velocity. Overall, our results offer new insights into the TBU toxicity to developing organisms, namely effects in AChE activity and hyperactivity, providing support data for future studies considering environmental risk assessment of this herbicide.

BDE-47, a flame retardant that is frequently detected in environmental compartments and human tissues, has been associated with various toxic effects. In turn, information about the effects of aluminum diethyl-phosphinate (ALPI), a halogen-free flame retardant from a newer generation, is limited. This study aims to assess and compare the toxicity of BDE-47 and ALPI to zebrafish by analyzing the tail coiling, locomotor, acetylcholinesterase activities, and oxidative stress biomarkers. At 3000 µg/L BDE-47, the coiling frequency increased at 26–27 h post-fertilization (hpf), but the burst activity (%) and mean burst duration (s) did not change significantly. Here, we considered that the increased coiling frequency is a slight neurotoxic effect because locomotor activity was impaired at 144 hpf and 300 µg/L BDE-47. Moreover, we hypothesized that oxidative stress could be involved in the BDE-47 toxicity mechanisms. In contrast, only at 30,000 µg/L did ALPI increase the catalase activity, while the motor behavior during different developmental stages remained unaffected. On the basis of these findings, BDE-47 is more toxic than ALPI.

Reactive oxygen species (ROS) are important second messengers in many metabolic processes and signaling pathways. Disruption of the balance between ROS generation and antioxidant defenses results in the overproduction of ROS and subsequent oxidative damage to biomolecules and cellular components that disturb cellular function. Oxidative stress contributes to the initiation and progression of many liver pathologies such as ischemia-reperfusion injury (LIRI), non-alcoholic fatty liver disease (NAFLD), and hepatocellular carcinoma (HCC). Therefore, controlling ROS production is an attractive therapeutic strategy in relation to their treatment. In recent years, increasing evidence has supported the therapeutic effects of polyphenols on liver injury via the regulation of ROS levels. In the current review, we summarize the effects of polyphenols, such as quercetin, resveratrol, and curcumin, on oxidative damage during conditions that induce liver injury, such as LIRI, NAFLD, and HCC.

Retene is a polycyclic aromatic hydrocarbon (PAH) emitted mainly by biomass combustion, and despite its ubiquity in atmospheric particulate matter (PM), studies concerning its potential hazard to human health are still incipient. In this study, the cytotoxicity and genotoxicity of retene were investigated in human HepG2 liver cells. Our data showed that retene had minimal effect on cell viability, but induced DNA strand breaks, micronuclei formation, and reactive oxygen species (ROS) formation in a dose- and time-dependent manner. Stronger effects were observed at earlier time points than at longer, indicating transient genotoxicity. Retene activated phosphorylation of Checkpoint kinase 1 (Chk1), an indicator of replication stress and chromosomal instability, which was in accordance with increased formation of micronuclei. A protective effect of the antioxidant N-acetylcysteine (NAC) towards ROS generation and DNA damage signaling was observed, suggesting oxidative stress as a key mechanism of the observed genotoxic effects of retene in HepG2 cells. Altogether our results suggest that retene may contribute to the harmful effects caused by biomass burning PM and represent a potential hazard to human health.

In the context of a rising global temperature, biomass burning represents an increasing risk to human health, due to emissions of highly toxic substances such as polycyclic aromatic hydrocarbon (PAHs). Size-segregated particulate matter (PM) was collected in a region within the sugarcane belt of São Paulo state (Brazil), where biomass burning is still frequent, despite the phasing out of manual harvesting preceded by fire. The median of the total concentration of the 15 PAHs determined was 2.3 ± 1.8 ng m−3 (n = 19), where 63% of this content was in PM1.0. Concentrations of OPAHs and NPAHs were about an order of magnitude lower. PM2.5 collected in the dry season, when most of the fires occur, presented PAHs and OPAHs total concentrations three times higher than in the wet season, showing positive correlations with fire foci number and levoglucosan (a biomass burning marker). These results, added to the fact that biomass burning explained 65% of the data variance (PCA analysis), evidenced the importance of this practice as a source of PAHs and OPAHs to the regional atmosphere. Conversely, NPAHs appeared to be mainly derived from diesel-powered vehicles. The B[a]P equivalent concentration was estimated to be 4 times higher in the dry season than in the wet season, and was greatly increased during a local fire event. Cytotoxicity and genotoxicity of PM1.0 organic extracts were assessed using in vitro tests with human liver HepG2 cells. For both types of tests, significant toxicity was only observed for samples collected during the dry season. Persistent DNA damage that may have impaired the DNA repair system was also observed. The results indicated that there was a health risk associated with the air particulate mixture, mainly related to biomass burning, demonstrating the urgent need for better remediation actions to prevent the occurrence of burning events.