A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage

Cells are continually exposed to agents arising from the internal and external environments, which may damage DNA. This damage can cause aberrant cell ...

Florida International University

Endosulfan, the last remaining organochlorine pesticide, has been the subject of a number of international regulations and restriction/banning action plans worldwide. Occurrence of endosulfan residues in South Florida environments has been widely described in the literature for more than two decades. This work describes a selective, sensitive, and fast online solid-phase extraction (SPE) method coupled with liquid chromatography separation and tandem mass spectrometry (LC-MS/MS) for the determination of endosulfan isomers and endosulfan sulfate in water samples at low part per trillion levels with very little sample preparation. A negative atmospheric pressure chemical ionization source was carefully optimized to produce reproducible spectra of the target compounds with no adduct ion formation. Selected reaction monitoring transitions were monitored and quantitation was performed against a per-deuterated internal standard β-endosulfan (d4). The automated online SPE clean-up was performed using only 20 mL of untreated water sample prior to LC-MS/MS analysis. The method was capable of separating and quantifying endosulfan within a 24-min run using acetonitrile and water as mobile phases and presenting statistically calculated method detection limits of 3, 10, and 7 ng/L for endosulfan sulfate, α-endosulfan, and β-endosulfan, respectively. In addition, a QuEChERS method was successfully developed and applied for endosulfan determination in sediments/soils, floating and submerged algal mats, and small fish. Minimal matrix effects were observed in all matrices analyzed and recoveries for all analytes ranged from 50-144 %. The developed methodology was applied to monitor the occurrence and to assess the potential transport of endosulfan in the Loveland Slough watershed, an area adjacent to Everglades National Park showing long-term contamination with endosulfans.

Outcompeting GC for the detection of legacy chlorinated pesticides: online-SPE UPLC APCI/MSMS detection of endosulfans at part per trillion levels.

Polydimethylsiloxane (PDMS) is a polymer material with high absorptive properties increasingly used as a passive environmental sampler for persistent organic compounds. However, the partitioning behavior of hydrophobic chemicals to PDMS remains largely unknown. Organochlorines (OCs) and polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants of great concern due to their persistence and potential toxic effects on humans and animals. In this study, the affinity of 20 OCs and 25 PAHs for commercially available PDMS pellets was determined to assess their effectiveness as passive samplers. Experiments were conducted to estimate the absorption rates (k) and equilibrium concentrations, demonstrating that 16 OCs and 21 PAHs were efficiently absorbed by PDMS, while others remained dissolved in water. A model has been proposed to predict dissolved concentrations in water based on the Kow of the compound, suggesting that PDMS is a suitable passive sampler for these compounds.

Using Polydimethylsiloxane (PDMS) Pellets to Create an Absorption Model for the Determination of Equilibrium Concentrations of Dissolved Contaminants in the Aquatic Environment.

Polychlorinated biphenyls' (PCB) exposure has been reported to be associated with depressive symptoms, which is correlated to lower dopamine- (DA) and thyroxine-concentrations (T4). T4 is necessary for DA-synthesis and it binds to transthyretin (TTR) being transported into the brain. PCBs can displace T4 by binding to TTR itself, being transported into the brain and disturbing DA-synthesis, where depressive symptoms might occur. Consequently, the free T4-concentration (fT4) increases when PCBs bind to TTR. The interaction of PCBs with fT4 and its associations with the main DA metabolite, homovanillic acid (HVA), and depressive symptoms were investigated. In total, 116 participants (91.6% men) were investigated, who took part in three annual examinations (t1⁻t3) of the HELPcB health surveillance program. Blood was collected for measuring PCBs, hydroxy PCBs (OH-PCBs), and fT4 and urine for HVA. Depressive Symptoms were assessed with a standardized questionnaire. Interactions were tested cross-sectionally with multiple hierarchical regressions and longitudinally with mixed effect models. Related to HVA, an interaction was cross-sectionally found for lower-chlorinated PCBs (LPCBs) and dioxin-like PCBs (dlPCBs); longitudinally only for LPCBs. Related to depressive symptoms, the interaction was found for LPCBs, dlPCBs, and OH-PCBs; longitudinally again only for LPCBs. The results give first hints that a physiological process involving the thyroid and DA system is responsible for depressive symptoms after PCB exposure.

Depressive Symptoms After PCB Exposure: Hypotheses for Underlying Pathomechanisms via the Thyroid and Dopamine System.

Seabirds are suitable biomonitors for several persistent organic pollutants (POP), such as polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polycyclic aromatic hydrocarbons (PAHs), although scarce studies of PAHs in seabirds are available, especially in South American populations. Therefore, this study aimed to assess OCPs, PCBs and PAHs, through gas chromatography-electron capture detector (GC-ECD) and gas chromatography-mass spectrometry (GC-MS) analyses, in liver (n = 9) and muscle tissue (n = 13) from juvenile Magellanic penguins (Spheniscus magellanicus) found stranded on the coast of Rio de Janeiro, Southeastern Brazil. DDT-related compounds were the most frequently detected OCP, and 4,4'-dichlorodiphenyldichloroethylene (DDE), the main DDT metabolite found in penguin tissues. OCP concentrations in liver were two-fold higher than in muscle tissues. Compound specific ratios identified recent exposure of penguins to some OCPs as well as evidence of legacy pollution associated with industrial sources. The predominant PCB congeners were PCB 8/5, PCB 138/160 and PCB 153/132, with concentrations ranging from <LOQ and 1500 ng g dry weight. This study comprises one of the few PAHs reports in penguin tissues, and, although most compounds were detected at very low levels or below the limit of quantitation (LOQ), the concentrations reported herein were up to 100-fold higher than in previous studies in penguins. Therefore, considering penguin vulnerability to marine oil spills during migration routes, further assessments are required in different tissues in order to assess potential environmental health risks to these sentinel species.

Occurrence and tissue distribution of organochlorinated compounds and polycyclic aromatic hydrocarbons in Magellanic penguins (Spheniscus magellanicus) from the southeastern coast of Brazil.

The current coronavirus pandemic is leading to significant impacts on the planet, changing our way of life. Although the COVID-19 virus mechanisms of action and pathogenesis are still under extensive research, immune system effects are evident, leading, in many cases, to respiratory distress. Although apparent pollution reduction has been noticed by the population, environmental and human health impacts due to the increased use of plastic waste and disinfectants is concerning. One of the main routes of human exposure to pollutants is through drinking water. Thus, this point of view discusses some major contaminants in drinking water known to be immunotoxic, exploring sources and drinking water routes and emphasizing the known mechanisms of action that could likely compromise the effective immune response of humans, particularly raising concerns regarding people exposed to the COVID-19 virus. Based on a literature review, metals, plastic components, plasticizers, and per- and polyfluoroalkyl substances may display the potential to exacerbate COVID-19 respiratory symptoms, although epidemiological studies are still required to confirm the synergistic effects between these pollutants and the virus.

Drinking water pollutants may affect the immune system: concerns regarding COVID-19 health effects.

Environmental exposure risk to different xenobiotics, which can potentially alter the function of the endocrine system, remains a great health and safety concern for aquatic species and humans. Steroid hormones, pharmaceuticals and personal care products (PPCPs) have been identified as important aquatic contaminants due to their widespread occurrence in surface waters and their endocrine disrupting properties. Heavily populated areas in South Florida not served by municipal wastewater collection present an unexpected high risk of anthropogenic contaminants to nearby coastal systems through surface runoff and groundwater flow. Previous studies in South Florida have been largely concentrated on assessing the relevance of the fate and transport of inorganic nutrients, heavy metals and pesticides with regulatory criteria. Therefore, a significant gap exists in assessing occurrence, distribution and biological significance of the presence of human related organic contaminants in natural surface waters. In this study, we have developed a fast and sensitive online solid-phase extraction followed by liquid chromatography-high resolution mass spectrometry (SPE-LC-HRMS) method using a Q-Exactive system for the determination of the occurrence and distribution of selected wastewater tracers/indicators, recalcitrant PPCPs and steroid hormones in South Florida surface waters. Seasonal and spatial variations of these contaminants were monitored from 2017 to 2019. The presence of total coliforms and E. coli were also evaluated in order to further assess water quality. Correlations between hormones and anthropogenic tracers were explored to better elucidate the sources, pathways and exposure risks to these contaminants. Caffeine, sucralose, Diethyl-m-toluamide (DEET) and carbamazepine were frequently detected in the water samples, which is indicative of extensive wastewater intrusion impacting the surface water. Estrone (E1), 17-β-estradiol (E2), and 17-α-ethynylestradiol (EE2) levels found in surface water raises concern of potential endocrine disruption effects in the aquatic ecosystem. Hazard quotient has been calculated to identify areas with high ecological risks to aquatic organisms.

Understanding the occurrence and distribution of emerging pollutants and endocrine disruptors in sensitive coastal South Florida Ecosystems.

Lipids are excellent biomarkers for assessing coral stress, although staghorn coral data (Acropora cervicornis) is lacking. Lipid extraction is the most critical step in lipidomic assessments, usually performed using carcinogenic solvents. Efficient alternative using less toxic methods, such as the BUME method using butanol and methanol as extraction solvents, have not been applied to coral lipidomics evaluations. Thus, we aimed to develop a lipidomic approach to identify important coral health biomarkers by comparing different solvent mixtures in staghorn corals. Total lipid extraction was equivalent for both tested methods, but due to its efficiency in extracting polar lipids, the BUME method was chosen. It was then applied to different coral masses (0.33-1.00 g), resulting in non-significant differences concerning number of lipid classes and compounds. Therefore, this method can be successfully applied to coral assessments in a climate change context, with the added benefit of low sample masses, lessening coral sampling impacts.

Application of an Improved Chloroform-Free Lipid Extraction Method to Staghorn Coral (Acropora cervicornis) Lipidomics Assessments.

Per- and polyfluoroalkyl substances (PFAS) are persistent anthropogenic pollutants present in many environmental media worldwide due to their extensive uses in many industrial and commercial products combined with their high thermal and chemical stabilities. Its ubiquitous presence in surface and drinking water supply and significant adverse health effects observed in wildlife and humans, associated with its bioaccumulation potential, pose big concerns. In this study, we have developed and validated a semi-automated solid phase extraction (SPE) followed by liquid chromatography-mass spectrometry (LC-MS/MS) for the determination of legacy and emerging short-chain PFAS substitutes in surface and tap water at low parts-per-trillion (ppt) levels in South Florida environments. Surface waters from Biscayne Bay and adjacent canals (n = 15) and tap waters from different counties (Miami-Dade, Broward, and Palm Beach County) (n = 21) were collected between October 2020 (wet season) and February 2021 (dry season). Total PFAS concentrations up to 242 ng L (average of 168 ng L) were found in tap water from Grapeland Heights, which is the closest location to the Miami international airport that was sampled. The highest average total PFAS level of 106 ng L was observed in surface water from the Biscayne Canal C-8 for the wet and dry season. In general, average total PFAS was higher in tap water (86.3 ng L) than in surface waters (46.3 ng L), whereas the most predominant and frequently detected PFAS were PFBA, PFBS, PFPeA, PFHxA, PFHxS, PFOA and PFOS. PFAS levels found could represent a high human health risk, and ecological risk based on PFOS levels above recommended thresholds are also noted. Such knowledge on PFAS occurrence, distribution and sources in South Florida will provide essential information for local and regional regulatory agencies related to water quality, further facilitating the development of guidelines and procedures for PFAS pollution control and reduction in Florida.

Assessment of per- and polyfluoroalkyl substances in Biscayne Bay surface waters and tap waters from South Florida.

Per- and polyfluoroalkyl substances (PFAS) and phthalate esters (PAEs) are emerging contaminants of higher concern due to their wide industrial and commercial use, toxicity, and potential adverse health effects. In this study, we assessed PFAS and PAEs exposure in American oysters collected in three study sites in Florida, USA. Potential physiological effects of these contaminants were assessed by collecting oyster biometric data, calculating condition indices, and assessing oxidative stress levels in these individuals. Finally, a human health risk assessment was conducted based on the concentrations found in the consumable Tampa Bay (TB) oysters. All PFAS and PAEs compounds assessed in this study were detected in at least one oyster in all study sites. Among all locations, ΣPFAS concentration range was 0.611-134.78 ng·g and ΣPAEs <0.328-1021 ng·g. Despite the smaller size of Biscayne Bay (BB) oysters, they displayed the highest concentrations of most of the PFAS and PAEs compounds, which is likely associated with population size, and other sources in the area. Condition index (CI) III was smaller in BB oysters, likely indicating a stressed population. Even though BB oysters were the most affected individuals, Marco Island (MI) oysters displayed the highest levels of lipid peroxidation, which can also be associated with environmental factors and decreased food availability. Conversely, TB oysters exhibited the highest levels of hydrogen peroxide, likely indicating a better defense mechanism in TB oysters compared to MI oysters. The human health risk assessment for TB oysters indicated low risk from PFAS and PAEs exposure, but there is no reference dose for other compounds and the human diet is wider than only oysters. Therefore, the risk of contaminant exposure is likely higher. This study demonstrates the value of integrating data on contaminant exposure and physiological responses of bioindicator specimens to better understand how emerging contaminants are affecting marine wildlife.

Natalia S. Quinete

Department of Chemistry and Biochemistry,
Institute of Environment,
Department of Chemistry and Biochemistry, Institute of Environment

Florida International University

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage

Natalia S. Quinete

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Chemistry and Biochemistry,Institute of Environment,Department of Chemistry and Biochemistry, Institute of Environment

Florida International University

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage

Department of Chemistry and Biochemistry,
Institute of Environment,
Department of Chemistry and Biochemistry, Institute of Environment