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1.
Environ Sci Technol ; 55(18): 12223-12232, 2021 09 21.
Artigo em Inglês | MEDLINE | ID: mdl-34506112

RESUMO

The Clean Air Act requires the United States Environmental Protection Agency to review routinely the National Ambient Air Quality Standards, including fine particulate matter (PM2.5). A non-governmental Independent Particulate Matter Review Panel recently concluded that the current PM2.5 standards do not protect public health adequately and recommended revising the daily standard from 35 to 25-30 µg/m3 and the annual standard from 12 to 8-10 µg/m3. To assess the public health implications of adopting the PM2.5 standards proposed by the panel, the health benefits are quantified from their implementation based on both current (observed) and future (simulated) air quality data for California. The findings indicate that strengthening the standards would provide significant public health benefits valued at $42-$149 billion. Additionally, the stronger standards are shown to benefit environmental justice via health savings that are allocated more within environmentally and socioeconomically disadvantaged communities.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , Poluentes Atmosféricos/análise , Poluição do Ar/análise , California , Exposição Ambiental , Material Particulado/análise , Estados Unidos , United States Environmental Protection Agency
2.
Environ Health ; 20(1): 104, 2021 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-34535123

RESUMO

Toxic chemicals - "toxicants" - have been studied and regulated as single entities, and, carcinogens aside, almost all toxicants, single or mixed and however altered, have been thought harmless in very low doses or very weak concentrations. Yet much work in recent decades has shown that toxicants can injure wildlife, laboratory animals, and humans following exposures previously expected to be harmless. Additional work has shown that toxicants can act not only individually and cumulatively but also collectively and even synergistically and that they affect disadvantaged communities inordinately - and therefore, as argued by reformers, unjustly. As late as December 2016, the last full month before the inauguration of a president promising to rescind major environmental regulations, the United States federal environmental-health establishment, as led by the Environmental Protection Agency (EPA), had not developed coherent strategies to mitigate such risks, to alert the public to their plausibility, or to advise leadership in government and industry about their implications. To understand why, we examined archival materials, reviewed online databases, read internal industry communications, and interviewed experts. We confirmed that external constraints, statutory and judicial, had been in place prior to EPA's earliest interest in mixture toxicity, but we found no overt effort, certainly no successful effort, to loosen those constraints. We also found internal constraints: concerns that fully committing to the study of complex mixtures involving numerous toxicants would lead to methodological drift within the toxicological community and that trying to act on insights from such study could lead only to regulatory futility. Interaction of these constraints, external and internal, shielded the EPA by circumscribing its responsibilities and by impeding movement toward paradigmatic adjustment, but it also perpetuated scientifically dubious policies, such as those limiting the evaluation of commercial chemical formulations, including pesticide formulations, to only those ingredients said by their manufacturers to be active. In this context, regulators' disregard of synergism contrasted irreconcilably with biocide manufacturers' understanding that synergism enhanced lethality and patentability. In the end, an effective national response to mixture toxicity, cumulative risk, and environmental injustice did not emerge. In parallel, though, the National Institute of Environmental Health Sciences, which was less constrained, pursued with scientific investigation what the EPA had not pursued with regulatory action.


Assuntos
Política Ambiental/história , Poluentes Ambientais/toxicidade , Substâncias Perigosas/toxicidade , National Institute of Environmental Health Sciences (U.S.)/história , Medição de Risco/história , United States Environmental Protection Agency/história , Saúde Ambiental/história , Regulamentação Governamental , História do Século XX , História do Século XXI , Humanos , Justiça Social , Estados Unidos
3.
Artigo em Inglês | MEDLINE | ID: mdl-34360177

RESUMO

The United States Environmental Protection Agency monitors contaminants in drinking water and consolidates these results in the National Contaminant Occurrence Database. Our objective was to assess the co-occurrence of metal contaminants (total chromium, hexavalent chromium, molybdenum, vanadium, cobalt, and strontium) over the years 2013-2015. We used multilevel Tobit regression models with state and water system-level random intercepts to predict the geometric mean of each contaminant occurring in each public water system, and estimated the pairwise correlations of predicted water system-specific geometric means across contaminants. We found that the geometric means of vanadium and total chromium were positively correlated both in large public water systems (r = 0.45, p < 0.01) and in small public water systems (r = 0.47, p < 0.01). Further research may address the cumulative human health impacts of ingesting more than one contaminant in drinking water.


Assuntos
Água Potável , Poluentes Químicos da Água , Água Potável/análise , Humanos , Estados Unidos , United States Environmental Protection Agency , Água , Poluentes Químicos da Água/análise , Abastecimento de Água
5.
Artigo em Inglês | MEDLINE | ID: mdl-34205009

RESUMO

Extensive scholarship has demonstrated that communities of color, low-income communities, and Indigenous communities face greater environmental and health hazards compared to communities with more White or affluent people. Low-income, Indigenous, Black, and/or other populations of color are also more likely to lack access to health care facilities, healthy food, and adequate formal education opportunities. Despite the mountains of evidence that demonstrate the existence and significance of the elevated toxic social and environmental exposures experienced by these communities, the inclusion of these factors into chemical evaluations has been scarce. In this paper, we demonstrate a process built with publicly available data and simple geospatial techniques that could be utilized by the U.S. Environmental Protection Agency (USEPA) to incorporate cumulative approaches into risk assessments under the Toxic Substances Control Act. The use of these approaches, particularly as they relate to identifying potentially exposed and susceptible subpopulations, would help USEPA develop appropriate risk estimates and mitigation strategies to protect disproportionately burdened populations from the adverse effects of chemical exposures. By utilizing such approaches to inform risk evaluation and mitigation, USEPA can identify and protect those most burdened and impacted by toxic chemicals, and finally begin to close the gap of environmental health inequities.


Assuntos
Exposição Ambiental , Saúde Ambiental , Formaldeído , Substâncias Perigosas/toxicidade , Humanos , Medição de Risco , Estados Unidos , United States Environmental Protection Agency
6.
Sci Data ; 8(1): 169, 2021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34253739

RESUMO

The EPA developed the Adverse Outcome Pathway Database (AOP-DB) to better characterize adverse outcomes of toxicological interest that are relevant to human health and the environment. Here we present the most recent version of the EPA Adverse Outcome Pathway Database (AOP-DB), version 2. AOP-DB v.2 introduces several substantial updates, which include automated data pulls from the AOP-Wiki 2.0, the integration of tissue-gene network data, and human AOP-gene data by population, semantic mapping and SPARQL endpoint creation, in addition to the presentation of the first publicly available AOP-DB web user interface. Potential users of the data may investigate specific molecular targets of an AOP, the relation of those gene/protein targets to other AOPs, cross-species, pathway, or disease-AOP relationships, or frequencies of AOP-related functional variants in particular populations, for example. Version updates described herein help inform new testable hypotheses about the etiology and mechanisms underlying adverse outcomes of environmental and toxicological concern.


Assuntos
Rotas de Resultados Adversos , Bases de Dados Factuais , United States Environmental Protection Agency , Conjuntos de Dados como Assunto , Redes Reguladoras de Genes , Humanos , Estados Unidos
7.
Environ Int ; 155: 106701, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34146765

RESUMO

Bisphenol A diglycidyl ether (BADGE)-based epoxy resin is one of the most widely used epoxy resins with an annual production amount of several million tons. Compared with all other legacy or emerging organic compounds, BADGE is special due to its toxicity and high reactivity in the environment. More and more studies are available on its analytical methods, occurrence, transformation and toxicity. Here, we provided a comprehensive review of the current BADGE-related studies, with focus on its production, application, available analytical methods, occurrences in the environment and human specimen, abiotic and biotic transformation, as well as the in vitro and in vivo toxicities. The available data show that BADGE and its derivatives are ubiquitous environmental chemicals and often well detected in human specimens. For their analysis, a water-free sample pretreatment should be considered to avoid hydrolysis. Additionally, their complex reactions with endogenous metabolites are areas of great interest. To date, the monitoring and further understanding of their transport and fate in the environment are still quite lacking, comparing with its analogues bisphenol A (BPA) and bisphenol S (BPS). In terms of toxicity, the summary of its current studies and Environmental Protection Agency (EPA) ToxCast toxicity database suggests BADGE might be an endocrine disruptor, though more detailed evidence is still needed to confirm this hypothesis in in vivo animal models. Future study of BADGE should focus on its metabolic transformation, reaction with protein and validation of its role as an endocrine disruptor. We believe that the elucidation of BADGEs can greatly enhance our understandings of those reactive compounds in the environment and human.


Assuntos
Disruptores Endócrinos , Poluentes Ambientais , Compostos Benzidrílicos/análise , Compostos Benzidrílicos/toxicidade , Poluentes Ambientais/toxicidade , Compostos de Epóxi , Humanos , Estados Unidos , United States Environmental Protection Agency
8.
Environ Pollut ; 284: 117372, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34087668

RESUMO

With a rule published on 6 January 2021, the US Environmental Protection Agency (EPA) considers for the first time sub-threshold responses, abandoning the use of default dose-response models. This may affect worldwide scientific research, in terms of research design and methodology, and regulatory actions in China and other countries.


Assuntos
United States Environmental Protection Agency , China , Medição de Risco , Estados Unidos
9.
J Hazard Mater ; 419: 126361, 2021 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-34157464

RESUMO

Per and poly-fluoroalkyl substances (PFAS) have been recognized as contaminants of emerging concerns by the United States Environmental Protection Agency (US EPA) due to their environmental impact. Several advisory guidelines were proposed worldwide aimed at limiting their occurrences in the aquatic environments, especially for perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA). This review paper aims to provide a holistic review in the emerging area of PFAS research by summarizing the spatiotemporal variations in PFAS concentrations in surface water systems globally, highlighting the possible trends of occurrences of PFAS, and presenting potential human health impacts as a result of PFAS exposure through surface water matrices. From the data analysis in this study, occurrences of PFOA and PFOS in many surface water matrices were observed to be several folds higher than the US EPA health advisory level of 70 ng/L for lifetime exposure from drinking water. Direct discharge and atmospheric deposition were identified as primary sources of PFAS in surface water and cryosphere, respectively. While global efforts focused on limiting usages of long-chain PFAS such as PFOS and PFOA, the practices of using short-chain PFAS such as perfluorobutanoic acid (PFBA) and perfluorobutane sulfonic acid (PFBS) and PFAS alternatives increased substantially. These compounds are also potentially associated with adverse impacts on human health, animals and biota.


Assuntos
Ácidos Alcanossulfônicos , Água Potável , Fluorcarbonetos , Poluentes Químicos da Água , Ácidos Alcanossulfônicos/toxicidade , Animais , Água Potável/análise , Fluorcarbonetos/análise , Fluorcarbonetos/toxicidade , Humanos , Ácidos Sulfônicos/toxicidade , Estados Unidos , United States Environmental Protection Agency , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade
10.
Environ Res ; 201: 111551, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34192556

RESUMO

The color effluent discharged by alcohol distilleries comprises very high pollution loads due to the plethora of refractory chemicals even after anaerobic treatment and causing adverse effects to the environment. The present study aimed to examine the phytotoxic, cytotoxic, and genotoxic potential of the identified refractory organic and inorganic pollutants discharged in bio-methanated distillery effluent (BMDE). Physico-chemical analyses revealed that BMDE retains high BOD, COD, TDS along with heavy metals like Fe (572.64 mg L-1), Mn (4.269 mg L-1), Cd (1.631 mg L-1), Zn (2.547 mg L-1), Pb (1.262 mg L-1), (Cr 1.257 mg L-1), and Ni (0.781 mg L-1) beyond the permissible limits for effluent discharge. GC-MS analysis revelaed the presence of hexadecanoic acid, TMS ester; octadecanoic acid, TMS ester; 2,3 bis[(TMS)oxy]propyl ester; stigmasterol TMS ether; ß-sitosterol TMS ester; hexacosanoic acid; and tetradecanoic acid, TMS ester as major refractory organic pollutants, which are listed as potential endocrine disruptor chemicals (EDCs) as per USEPA. Furthermore, phytotoxicity assessment with Phaseolus aureus L. showed the toxic nature of BMDE as it inhibited various seedling growth parameters, seed germination, and suppression of α-amylase activity in seed germination experiment. Moreover, genotoxicity and cytotoxicity evaluation of the discharged BMDE evidenced in root-tip meristematic cells of Allium cepa L. where chromosomal aberration such as disturbed metaphase, c-mitosis, laggard chromosomes, sticky chromosomes, prolonged prophase, polyploid cells, and apoptotic bodies etc. were observed. Thus, this study's results suggested that BMDE discharged without adequate treatment poses potential risk to environment and may cause a variety of serious health threats in living beings upon exposure.


Assuntos
Poluentes Ambientais , Phaseolus , Dano ao DNA , Cebolas , Estados Unidos , United States Environmental Protection Agency
12.
Environ Int ; 154: 106566, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-33934018

RESUMO

For the past six decades, human health risk assessment of chemicals has relied on in vivo data from human epidemiological and experimental animal toxicological studies to inform the derivation of non-cancer toxicity values. The ongoing evolution of this risk assessment paradigm in an environmental landscape of data-poor chemicals has highlighted the need to develop and implement non-testing methods, so-called New Approach Methodologies (NAMs). NAMs include a growing number of in silico and in vitro data streams designed to inform hazard properties of chemicals, including kinetics and dynamics at different levels of biological organization, environmental fate and transport, and exposure. NAMs provide a fit-for-purpose science-basis for human hazard and risk characterization of chemicals ranging from data-gap filling applications to broad evidence-based decision-making. Systematic assembly and delivery of empirical and predicted data for chemicals are paramount to advancing chemical evaluation, and software tools serve an essential role in delivering these data to the scientific community. The CompTox Chemicals Dashboard (from here on referred to as the "Dashboard") is one such tool and is a publicly available web-based application developed by the US Environmental Protection Agency to provide access to chemistry, toxicity and exposure information for ~900,000 chemicals. The Dashboard is increasingly becoming a valuable resource for assessors tasked with the evaluation of potential human health risks associated with chemical exposures. In this context, the significant amount of information present in the Dashboard facilitates: 1) assembly of information on physicochemical properties and environmental fate and transport and exposure parameters and metrics; 2) identification of cancer and non-cancer health effects from extant human and experimental animal studies in the public domain and/or information not available in the public domain (i.e., "grey literature"); 3) systematic literature searching and review for developing cancer and non-cancer hazard evidence bases; and 4) access to mechanistic information that can aid or augment the analysis of traditional toxicology evidence bases, or potentially, serve as the primary basis for informing hazard identification and dose-response when traditional bioassay data are lacking. Finally, in silico predictive tools developed to conduct structure-activity or read-across analyses are also available within the Dashboard. This practical tutorial is intended to address key questions from the human health risk assessment community dealing with chemicals in both food and in the environment. Perspectives for future development or refinement of the Dashboard highlight foreseen activities to further support the research and risk assessment community in cancer and non-cancer chemical evaluations.


Assuntos
United States Environmental Protection Agency , Animais , Simulação por Computador , Humanos , Medição de Risco , Estados Unidos
13.
J Chromatogr A ; 1647: 462131, 2021 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-33971520

RESUMO

Three modes of facilitating mini-scale-liquid-liquid-extraction (msLLE) prior to automated integration with full evaporation dynamic headspace (FEDHS) extraction were evaluated in this work. For msLLE, 1.2 mL of dichloromethane (DCM) was added to a conical-bottomed vial containing 7 mL of aqueous sample. The solution was then subjected to three different mixing modes, namely vortex-assistance (where a "whirlpool" was created in the solution), agitation-assistance (where the vial was rotated in circular motion) and quickMix-assistance (where the vial was shaken at a high speed). Vortex-assistance was performed manually while the other two modes were automated using a commercial autosampler. Following this, the DCM extract was transferred automatically to another vial and was then vaporized and sent through a Tenax TA sorbent tube in the FEDHS step. Due to the stronger π interaction between the sorbent and the analytes of interest, the analytes were selectively concentrated while the DCM vapor passed through unhampered. After FEDHS, the analytes were thoroughly desorbed into a gas chromatography-mass spectrometric system for analysis. The applicability of this procedure was validated in the extraction of six chlorinated benzenes (CBs) (1,2-dichlorobenzene, 1,3-dichlorobenzene, 1,4-dichlorobenzene, 1,2,4-trichlorobenzene, 1,2,4,5-tetrachlorobenzne and hexachlorobenzene) from aqueous samples. The quickMix-assisted msLLE-FEDHS approach achieved good absolute extraction recoveries (between 74.2% and 88.7%), low limits of detection (between 0.0006 and 0.0116 µg/L), good linearity (r2≥0.9920), good repeatability (between 1.9% and 8.4%, and good reproducibility (between 9.0% and 13.6%). It was found to be superior to the methods published by the United States Environmental Protection Agency. Five consecutive fully automated quickMix-assisted-msLLE-FEDHS-GC-MS runs spanned only ca. 4 hr.


Assuntos
Clorobenzenos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Extração Líquido-Líquido/métodos , Clorobenzenos/análise , Clorobenzenos/química , Clorobenzenos/isolamento & purificação , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos Testes , Estados Unidos , United States Environmental Protection Agency
14.
Int J Mol Sci ; 22(6)2021 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-33799614

RESUMO

The estrogen receptors α (ERα) are transcription factors involved in several physiological processes belonging to the nuclear receptors (NRs) protein family. Besides the endogenous ligands, several other chemicals are able to bind to those receptors. Among them are endocrine disrupting chemicals (EDCs) that can trigger toxicological pathways. Many studies have focused on predicting EDCs based on their ability to bind NRs; mainly, estrogen receptors (ER), thyroid hormones receptors (TR), androgen receptors (AR), glucocorticoid receptors (GR), and peroxisome proliferator-activated receptors gamma (PPARγ). In this work, we suggest a pipeline designed for the prediction of ERα binding activity. The flagged compounds can be further explored using experimental techniques to assess their potential to be EDCs. The pipeline is a combination of structure based (docking and pharmacophore models) and ligand based (pharmacophore models) methods. The models have been constructed using the Environmental Protection Agency (EPA) data encompassing a large number of structurally diverse compounds. A validation step was then achieved using two external databases: the NR-DBIND (Nuclear Receptors DataBase Including Negative Data) and the EADB (Estrogenic Activity DataBase). Different combination protocols were explored. Results showed that the combination of models performed better than each model taken individually. The consensus protocol that reached values of 0.81 and 0.54 for sensitivity and specificity, respectively, was the best suited for our toxicological study. Insights and recommendations were drawn to alleviate the screening quality of other projects focusing on ERα binding predictions.


Assuntos
Disruptores Endócrinos/química , Receptor alfa de Estrogênio/química , Simulação de Acoplamento Molecular , Sítios de Ligação , Bases de Dados de Compostos Químicos , Conjuntos de Dados como Assunto , Disruptores Endócrinos/classificação , Disruptores Endócrinos/metabolismo , Receptor alfa de Estrogênio/metabolismo , Humanos , Ligantes , Ligação Proteica , Projetos de Pesquisa , Sensibilidade e Especificidade , Relação Estrutura-Atividade , Estados Unidos , United States Environmental Protection Agency
15.
PLoS One ; 16(4): e0249913, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33857202

RESUMO

Vaporization of benzene, toluene, ethylbenzene, and xylene (BTEX) compounds pollutes the air and causes health hazards at gasoline stations. This study revealed the risk of BTEX exposure according to the hazardous area classification at gasoline stations. The risk assessment of gasoline workers from a representative group of 47 stations, which followed the United States Environmental Protection Agency-IRIS method of assessing BTEX exposure, was expressed as the hazard index (HI). A result of matrix multipliers of the hazardous exposure index and fire possibility from flammable gas classified hazardous area-I and area-II at the fuel dispensers. BTEX concentrations were actively sampled in ambient air and a flammable gas detector was used to measure the flammability level. Results showed that the BTEX concentrations from ambient air monitoring were in the range of 0.1-136.9, 8.1-406.0, 0.8-24.1 and 0.4-105.5 ppb for benzene, toluene, ethylbenzene, and xylene, respectively, which exceeded the NIOSH exposure limit of 100 ppb of benzene concentration. The risk assessment indicated that five stations reached an unacceptable risk of worker exposure to BTEX (HI>1), which correlated with the numbers of gasoline dispensers and daily gasoline sold. The risk matrix classified hazardous area-I at 4 meters and hazardous area-II at 4-8 meters in radius around the fuel dispensers. This study revealed the hazardous areas at gasoline stations and suggests that entrepreneurs must strictly control the safety operation practice of workers, install vapor recovery systems on dispenser nozzles to control BTEX vaporization and keep the hazardous areas clear of fire ignition sources within an eight-meter radius of the dispensers.


Assuntos
Poluentes Ocupacionais do Ar/análise , Gasolina , Substâncias Perigosas/análise , Exposição por Inalação/estatística & dados numéricos , Veículos Automotores , Exposição Ocupacional/estatística & dados numéricos , Gestão da Segurança/estatística & dados numéricos , Derivados de Benzeno/análise , Humanos , National Institute for Occupational Safety and Health, U.S./normas , Medição de Risco , Estados Unidos , United States Environmental Protection Agency/normas
17.
Artigo em Inglês | MEDLINE | ID: mdl-33804855

RESUMO

The development of high-throughput screening methodologies may decrease the need for laboratory animals for toxicity testing. Here, we investigate the potential of assessing immunotoxicity with high-throughput screening data from the U.S. Environmental Protection Agency ToxCast program. As case studies, we analyzed the most common chemicals added to food as well as per- and polyfluoroalkyl substances (PFAS) shown to migrate to food from packaging materials or processing equipment. The antioxidant preservative tert-butylhydroquinone (TBHQ) showed activity both in ToxCast assays and in classical immunological assays, suggesting that it may affect the immune response in people. From the PFAS group, we identified eight substances that can migrate from food contact materials and have ToxCast data. In epidemiological and toxicological studies, PFAS suppress the immune system and decrease the response to vaccination. However, most PFAS show weak or no activity in immune-related ToxCast assays. This lack of concordance between toxicological and high-throughput data for common PFAS indicates the current limitations of in vitro screening for analyzing immunotoxicity. High-throughput in vitro assays show promise for providing mechanistic data relevant for immune risk assessment. In contrast, the lack of immune-specific activity in the existing high-throughput assays cannot validate the safety of a chemical for the immune system.


Assuntos
Ensaios de Triagem em Larga Escala , Testes de Toxicidade , Animais , Alimentos , Medição de Risco , Estados Unidos , United States Environmental Protection Agency
18.
Environ Manage ; 67(6): 1017-1028, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33837478

RESUMO

Research and development has been a key part of the foundation for improvements in US air quality since the establishment of the Environmental Protection Agency (EPA) 50 years ago. Although the scientific accomplishments and advances over the course of EPA's history are often overshadowed by policy debates, much of the air pollution science and engineering we now consider to be routine did not exist when EPA was established. Many of the advances in air pollutant measurement, monitoring, modeling, and control were developed by EPA researchers or supported by EPA programs. The technical foundation built during EPA's early years has since given the Agency the scientific ability to respond quickly and effectively to unexpected and emerging issues. Equally important, EPA also developed approaches to conducting and presenting science in policy settings to ensure that the science was as objective and complete as possible and was communicated effectively. A look back at some of the accomplishments of EPA scientists and engineers provides a reminder that the cumulative effect of continual, incremental advances can result in large and lasting benefits to society.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , Poluentes Atmosféricos/análise , Poluição do Ar/análise , Poluição do Ar/prevenção & controle , Estados Unidos , United States Environmental Protection Agency
19.
J Microbiol Methods ; 184: 106206, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33766607

RESUMO

Droplet digital polymerase chain reaction (ddPCR) was evaluated for the detection of fecal indicator bacteria (FIB), Enterococcus spp., in San Diego County beach water samples collected under diverse conditions, from multiple pollution sources, as part of regulatory monitoring activities over 20 months. Two US EPA-approved methods, qPCR (EPA 1609.1) and Enterolert (SM9230D), were used as reference comparator methods. A total of 361 samples were assayed by both ddPCR and qPCR and yielded an acceptable Index of Agreement (IA) of 0.89, based on EPA Site-Specific analysis guidelines. A Pearson's correlation coefficient of r = 0.87 (p < 0.001), further indicated a strong relationship between the methods results. From the 361 samples, 185 split samples with ddPCR and Enterolert values within the limits of quantification, were used as a 'training' data set to derive an intrinsic copy number equation (ICE) for scaling ddPCR gene copy number to Enterolert most probable number (MPN). Of the 1993 samples that comprised the complete 'test' data set assayed by ddPCR and Enterolert, 1086 generated results that fell within the limits of quantification for Enterolert and yielded an overall IA of 0.64. Re-analysis using median as a measure of central tendency to account for significant skewing of Enterolert data yielded an IA of 0.72. Beach grouping-specific IA values ranged from 0.63 to 0.93. Pearson's correlation coefficient, r, ranged from 0.13 to 0.94 within beach groupings and generated a combined value of 0.60 for all groupings. Using the ICE, a ddPCR advisory threshold of 1413 DNA copy number/100 mL was empirically determined to be the equivalent to the California Enterolert beach action threshold of 104 MPN/100 mL, based on comparison with all 1993 paired ddPCR and Enterolert results. Using the 1413 DNA copy number/100 mL as a beach action threshold for ddPCR resulted in a 90.4% agreement with Enterolert (6.0% false negative and 3.7% false positive). Together these findings support the conclusion that ddPCR readouts align closely with Enterolert MPN for identifying FIB exceedance levels of Enterococcus spp. in coastal waters of San Diego, CA.


Assuntos
Bactérias/genética , Bactérias/isolamento & purificação , Monitoramento Ambiental/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Água do Mar/microbiologia , Bactérias/crescimento & desenvolvimento , Fezes/microbiologia , Humanos , Água do Mar/química , Estados Unidos , United States Environmental Protection Agency , Poluição da Água/análise , Qualidade da Água
20.
J Microbiol Methods ; 184: 106186, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33766609

RESUMO

Fecal pollution remains a challenge for water quality managers at Great Lakes and inland recreational beaches. The fecal indicator of choice at these beaches is typically Escherichia coli (E. coli), determined by culture-based methods that require over 18 h to obtain results. Researchers at the United States Environmental Protection Agency (EPA) have developed a rapid E. coli qPCR methodology (EPA Draft Method C) that can provide same-day results for improving public health protection with demonstrated sensitivity, specificity, and data acceptance criteria. However, limited information is currently available to compare the occurrence of E. coli determined by cultivation and by EPA Draft Method C (Method C). This study provides a large-scale data collection effort to compare the occurrence of E. coli determined by these alternative methods at more than 100 Michigan recreational beach and other sites using the complete set of quantitative data pairings and selected subsets of the data and sites meeting various eligibility requirements. Simple linear regression analyses of composite (pooled) data indicated a correlation between results of the E. coli monitoring approaches for each of the multi-site datasets as evidenced by Pearson R-squared values ranging from 0.452 to 0.641. Theoretical Method C threshold values, expressed as mean log10 target gene copies per reaction, that corresponded to an established E. coli culture method water quality standard of 300 MPN or CFU /100 mL varied only from 1.817 to 1.908 for the different datasets using this model. Different modeling and derivation approaches that incorporated within and between-site variability in the estimates also gave Method C threshold values in this range but only when relatively well-correlated datasets were used to minimize the error. A hypothetical exercise to evaluate the frequency of water impairments based on theoretical qPCR thresholds corresponding to the E. coli water quality standard for culture methods suggested that the methods may provide the same beach notification outcomes over 90% of the time with Method C results differing from culture method results that indicated acceptable and unacceptable water quality at overall rates of 1.9% and 6.6%, respectively. Results from this study provide useful information about the relationships between E. coli determined by culture and qPCR methods across many diverse freshwater sites and should facilitate efforts to implement qPCR-based E. coli detection for rapid recreational water quality monitoring on a large scale in the State of Michigan.


Assuntos
Contagem de Colônia Microbiana/métodos , Monitoramento Ambiental/métodos , Escherichia coli/isolamento & purificação , Lagos/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Michigan , Estados Unidos , United States Environmental Protection Agency , Qualidade da Água
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