A state-of-the-science review of analytical methods for urinary dialkylphosphate metabolites in the assessment of exposure to organophosphate pesticides: From 2000 to 2022.

The general population and workers are exposed to organophosphate insecticides, one of the leading chemical classes of pesticides used in rural and urban areas. This paper aims to conduct an integrative review of the most used analytical methods for identifying and quantifying dialkylphosphate-which are metabolites of organophosphate insecticides-in the urine of exposed workers, discussing their advantages, limitations and applicability. Searches utilized the PubMed, the Scientific Electronic Library Online and the Brazilian Digital Library of Theses and Dissertations databases between 2000 and 2021. Twenty-five studies were selected. The extraction methods most used were liquid-liquid extraction (LLE) (36%) and solid-phase extraction (SPE) (36%), with the SPE being more economical in terms of time and amount of solvents needed, and presenting the best percentage of recovery of analytes, when compared with LLE. Nineteen studies (76%) used the gas chromatography method of separation, and among these, 12 records (63%) indicated mass spectrometry used as a detection technology (analyzer). Studies demonstrate that dialkylphosphates are sensitive and representative exposure biomarkers for environmental and occupational organophosphate exposure.

Hematological Changes in Gas Station Workers.

(1) Background: Benzene, toluene, and xylene isomers (BTX) are present in gasoline. Exposure to benzene may lead to the appearance of a series of signs, symptoms, and complications, which are characterized by benzene poisoning, which is an occupational disease. This study evaluated the presence of signs and symptoms related to occupational exposure and whether occupational exposure to BTX is associated with the development of hematological changes. (2) Material and Methods: This cross-sectional epidemiological study included 542 participants, in which 324 were gas station workers (GSWs) and 218 were office workers (OWs) with no occupational exposure to benzene. To characterize the type of exposure (exposed and not exposed), trans,trans-Muconic acid (tt-MA), Hippuric acid (HA), and Methylhippuric acid (MHA) were used as exposure biomarkers. The tt-MA analysis revealed that the GSWs had 0.29 mg/g of urinary creatinine and the OWs had 0.13 mg/g of urinary creatinine. For HA, the GSWs presented 0.49 g/g of creatinine while the OWs presented 0.07. MHA analysis revealed that the GSWs had 1.57 g/g creatinine and the OWs had 0.01 g/g creatinine. Occupation habits and clinical symptoms were collected by questionnaire and blood samples were analyzed for hematological parameters. The persistence of hematological changes was evaluated with three serial blood collections every 15 days followed by laboratory hematological analysis. A descriptive analysis by the Chi-square test method was performed to evaluate the association between occupational exposure to fuels and the occurrence of changes in hematological parameters. (3) Results: In the GSWs, the most described signs and symptoms were somnolence (45.1%), headache (38.3%), dizziness (27.5%), tingling (25.4%), and involuntary movement (25%). Twenty GSWs that presented hematological alterations performed serial collections fifteen days apart. In addition, these workers presented total leukocyte counts above the upper limit and lymphocyte counts close to the lower limit. Leukocytosis and lymphopenia are hematological alterations present in chronic benzene poisoning. (4) Conclusions: The results found an initial change in different hematological parameters routinely used in clinics to evaluate health conditions. These findings reveal the importance of valuing clinical changes, even in the absence of disease, during the health monitoring of gas station workers and other groups that share the same space.

Evaluation of the developmental toxicity of solvents, metals, drugs, and industrial chemicals using a freshwater snail (Biomphalaria glabrata) assay.

A freshwater snail assay was employed to assess the embryotoxicity of solvents including acetone, methanol, ethanol, isopropanol, dimethyl-sulfoxide, glycerin, metals/metalloids including mercuric chloride (HgCl2), cadmium chloride (CdCl2,), antimony salts Sb+3 and Sb+5, drugs including colchicine, hydroxyurea, cyclophosphamide, an industrial chemical sodium azide (SA), an anionic surfactant dodecyl sodium sulfate-(DSS), H2O2 and sodium chloride (NaCl). The assay consists of exposing Biomphalaria glabrata egg masses (EM) to the substances for 96-hr and following up embryo/snail development for lethality, abnormal morphology (teratogenicity), and day of hatching up to day 10 or 14 after spawning. Based upon concentration-response relationships, LC50%s (embryolethality), EC50%s (teratogenicity) and IC50%s (hatching retardation) and 95%CIs were determined for tested chemicals. The LOECs indicated that HgCl2 (37 nM) and CdCl2 (140 nM) are potent embryotoxic agents in snails. Teratogenic indices (TI = LC50/EC50) for almost all tested chemicals were lower than or close to unity suggesting that these compounds were not teratogenic in this assay. The snail assay may be adequately performed in a cost-effective standardized protocol which enables testing a number of environmental chemicals over a broad concentration range. The snail assay needs to undergo further validation to be recognized for an internationally harmonized hazard identification in ecotoxicity risk assessment.

Risks related to the domestic laundering of filling station attendant uniforms: advances and uncertainties.

Gasoline is the most common transportation fuel in Brazil, with up to 1% of benzene. Benzene is a health-damaging organic solvent that is extensively used worldwide, being classified as a human carcinogen by the International Agency for Research on Cancer (Group 1). Many workers at filling stations are exposed to benzene, present in gasoline. The main routes of exposure and absorption of benzene are inhalation, oral, and dermal routes. The penetration and maintenance of benzene and other chemicals on personal protective equipment and clothing of workers who manipulate these solvents may increase their levels of exposure and offer risks to their family members, since contaminated clothing is laundered at their homes, which goes against legislative framework (Annex 2, Regulatory Standard No. 9). This way, the objective of this work was to discuss the importance of periodic changes and cleaning of filling station attendant uniforms performed by employers as preventive measures against the deterioration of the health of workers and their family members. We performed a narrative review; no systematic criteria were used in the search for national and international studies. After critical reading, we observed a lack of consistent data on this theme. Laundering of uniforms worn by filling station attendants should take place outside the domestic environment in order to protect the health of workers and their family members and avoid possible cross-contamination.

Gasoline-station workers in Brazil: Benzene exposure; Genotoxic and immunotoxic effects.

Chronic exposure to benzene is a risk factor for hematological malignancies. Gasoline-station workers are exposed to benzene in gasoline, via both inhalation and dermal contact (attendants and managers) or inhalation (workers in the on-site convenience stores and offices). We have studied the exposure of these workers to benzene and the resulting genotoxic and immunotoxic effects. Levels of urinary trans, trans-muconic acid were higher among gasoline-station workers than among office workers with no known exposure to benzene (comparison group). Among the exposed workers, we observed statistically significant biological effects, including elevated DNA damage (comet assay); higher frequencies of micronuclei and nuclear buds (CBMN assay); lower levels of T-helper lymphocytes and naive Th lymphocytes; lower CD4 / CD8 ratio; and higher levels of NK cells and memory Th lymphocytes. Both groups of exposed workers (inhalation and inhalation + dermal routes) showed similar genotoxic and immunotoxic effects.

Occurrence of green tobacco sickness and associated factors in farmers residing in Dom Feliciano Municipality, Rio Grande do Sul State, Southern Region of Brazil.

This study aimed to evaluate the occurrence of the green tobacco sickness (GTS) and its associated factors in tobacco familiar farmers residing in Dom Feliciano, Rio Grande do Sul State, Brazil. A cross-sectional study was conducted evaluating the sickness in 354 small tobacco farmers, between October 2011 and March 2012. The urinary concentration of cotinine, a biomarker of exposure to nicotine, was determined during the tobacco harvest period. Subjects presenting cotinine urinary levels ≥ 50ng/mL, that had contact with tobacco leaves up to 48 hours before the sample collection and reported at least one disease symptom were deemed as cases. A non-conditional logistic analysis was performed to evaluate the association between GTS and the population characteristics. A total of 122 (34.5%) cases were identified, with 39% of them being smokers and 61% being males. The median cotinine urinary concentrations were 75.6ng/mL (74.1ng/mg of creatinine) for non-cases and 755.8ng/mL (632.1ng/mg of creatinine) for the cases (p-value ≤ 0.01). The multivariate analysis showed a positive association between GTS and sunlight exposure time, exposure to pesticides, worse health status, and inverse association with wood cultivation. This study presented a high GTS prevalence and suggest that the use of urinary cotinine is a significant biomarker to determine GTS cases, influencing in the distribution by sex. Once the tobacco production involves health hazards, is important to implement measures to prevent the harm caused to tobacco farmers, as set in articles 17 and 18 of the World Health Organization Framework Convention on Tobacco Control.

Analysis of Benzene Exposure in Gas Station Workers Using Trans,Trans-Muconic Acid.

In Brazil, gas station workers are occupationally exposed to the benzene present in gasoline. Brazilian law indicates the use of trans,trans-muconic acid(t,t-MA) as a biomarker of benzene exposure. The aim of this study was to evaluate the level of exposure to benzene in gas station workers, through the quantification of t,t-MA present in urine. A total number of 269 gas station workers divided into 179 filling station attendants exposed by inhalation and dermal route and 90 convenience store workers exposed only by inhalation were included. A control group was formed by 100 office workers, without occupational exposure to benzene. The urinary levels of t,t-MA were evaluated by HPLC with a UV detector. Gas station workers showed higher mean values of t,t-MA (0.204 mg/g creatinine; 95% CI 0.170-0.237) than office workers (0.126 mg/g creatinine; 95% CI 0.0817-0.1693). T,t-MA levels were higher in convenience store workers exposed to gasoline only by inhalation (0.221 mg/g creatinine; 95% CI 0.160-0.282), than in those exposed to gasoline by inhalation and dermal route-filling station attendants (0.195 mg/g creatinine; 95% CI 0.155-0.235). Gas station workers with a higher level of t,t-MA had epistaxis. T,t-MA values were higher in the Downtown (0.15 mg/g creatinine) region's workers than in the more affluent South Zone region's workers (0.07 mg/g creatinine). Smoking habits influenced the urinary t,t-MA values, while the frequency of consumption of industrialized and frozen foods showed no influence.

Occurrence of green tobacco sickness and associated factors in farmers residing in Dom Feliciano Municipality, Rio Grande do Sul State, Southern Region of Brazil / Ocorrência da doença da folha verde do tabaco e fatores associados entre fumicultores no Município de Dom Feliciano, Rio Grande do Sul, Região Sul do Brasil / Ocurrencia de la enfermedad del tabaco verde y factores asociados en agricultores que residen en el Municipio de Dom Feliciano, Estado de Rio Grande do Sul, Región Sur de Brasil

Abstract: This study aimed to evaluate the occurrence of the green tobacco sickness (GTS) and its associated factors in tobacco familiar farmers residing in Dom Feliciano, Rio Grande do Sul State, Brazil. A cross-sectional study was conducted evaluating the sickness in 354 small tobacco farmers, between October 2011 and March 2012. The urinary concentration of cotinine, a biomarker of exposure to nicotine, was determined during the tobacco harvest period. Subjects presenting cotinine urinary levels ≥ 50ng/mL, that had contact with tobacco leaves up to 48 hours before the sample collection and reported at least one disease symptom were deemed as cases. A non-conditional logistic analysis was performed to evaluate the association between GTS and the population characteristics. A total of 122 (34.5%) cases were identified, with 39% of them being smokers and 61% being males. The median cotinine urinary concentrations were 75.6ng/mL (74.1ng/mg of creatinine) for non-cases and 755.8ng/mL (632.1ng/mg of creatinine) for the cases (p-value ≤ 0.01). The multivariate analysis showed a positive association between GTS and sunlight exposure time, exposure to pesticides, worse health status, and inverse association with wood cultivation. This study presented a high GTS prevalence and suggest that the use of urinary cotinine is a significant biomarker to determine GTS cases, influencing in the distribution by sex. Once the tobacco production involves health hazards, is important to implement measures to prevent the harm caused to tobacco farmers, as set in articles 17 and 18 of the World Health Organization Framework Convention on Tobacco Control.

Resumo: O estudo teve como objetivo avaliar a ocorrência de doença da folha verde (DFV) e fatores associados entre fumicultores familiares no Município de Dom Feliciano, Estado do Rio Grande do Sul, Brasil. Foi realizado um estudo transversal sobre DFV em 354 pequenos produtores de tabaco, entre outubro de 2011 e março de 2012. Foi medida a concentração urinária da cotinina, um biomarcador da exposição à nicotina, durante a temporada de colheita do tabaco. Os casos de DFV foram definidos como indivíduos com níveis urinários de cotinina ≥ 50ng/mL, que tiveram contato com folhas de tabaco em até 48 horas antes da coleta da amostra sanguínea e que relatavam pelo menos um sintoma da doença. Foi realizada uma análise logística não condicional para avaliar a associação entre DFV e as características da população. Foram identificados um total de 122 (34,5%) casos da doença, entre os quais 39% eram fumantes e 61% eram do sexo masculino. As concentrações urinárias medianas de cotinina foram 75,6ng/mL (74,1ng/mg de creatinina) para não-casos e 755,8ng/mL (632,1ng/mg de creatinina) para os casos (valor de p ≤ 0,01). A análise multivariada mostrou uma associação positiva entre DFV e tempo de exposição à luz solar, exposição a pesticidas e pior estado de saúde geral, e uma associação inversa com cultivo de lenha. O estudo identificou alta prevalência de DFV e sugere o uso da cotinina como biomarcador significativo para identificar os casos de DFV, influenciando a distribuição por sexo. Uma vez que a fumicultura envolve danos à saúde, é importante implementar medidas para prevenir os impactos para os fumicultores, de acordo com os artigos 17 e 18 da Convenção-Quadro para o Controle do Tabaco da Organização Mundial da Saúde.

Resumen: Este estudio visó evaluar la ocurrencia de la enfermedad del tabaco verde (GTS por sus siglas en inglés) y sus factores asociados en pequeños agricultores de tabaco, que residen en Dom Feliciano, Rio Grande do Sul, Brasil. Se realizó un estudio transversal, evaluando la enfermedad del tabaco verde, en 354 pequeños agricultores de tabaco, entre octubre de 2011 y marzo 2012. La concentración urinaria de cotinina, un biomarcador de la exposición a la nicotina, se determinó durante el periodo de cosecha del tabaco. Los individuos que presentan niveles de cotinina urinarios ≥ 50ng/mL, que tuvieron contacto con hojas de tabaco hasta 48 horas antes de la recogida de la muestra, e informaron de al menos un síntoma de la enfermedad, fueron considerados casos. Se realizó un análisis logístico no condicional para evaluar la asociación entre GTS y las características de la población. Se identificaron un total de 122 (34,5%) casos, siendo un 39% de ellos fumadores y un 61% hombres. Las concentraciones medias de cotinina urinaria fueron 75,6ng/mL (74,1ng/mg de creatinina) para los no casos y 755,8ng/mL (632,1ng/mg de creatinina) para los casos (valor de p ≤ 0,01). El análisis multivariado mostró una asociación positiva entre el GTS y las horas de exposición a la luz solar, exposición a los pesticidas, un estatus peor de salud, y la asociación inversa con la cultura maderera. Este estudio presentó una alta prevalencia de GTS y sugiere que el uso de la cotinina urinaria es un biomarcador significativo para identificar casos de GTS, influenciando en la distribución por sexo. Debido a que la producción de tabaco implica perjuicios para la salud, es importante implementar medidas para prevenirlos en los agricultores del tabaco, como está estipulado en los artículos 17 y 18 del Convenio Marco para el Control del Tabaco de la Organización Mundial de la Salud.

Estudo dos efeitos embriotóxicos de xenobióticos em biomphalaria glabrata / Study of the embryotoxics effect of xenobiotics in glabrata biomphalaria

(...) O objetivo deste estudo foi avaliar, em B.glabrata, o efeito embriotóxico de alguns solventes, i.e., acetona, etanol e do dimetilsulfóxido (DMSO) e de substâncias que foram genotóxicas, carcinogênicas e/ou teratogênicas em outros ensaios in vitro e in vivo, i.e., azida sódica (AS), ciclofosfamida (CP), cloreto de cádmio (CdCl2), colchicina (COL) e peróxido de hidrogênio (H2O2). Desovas de B. glabrata (< 15h após a postura, estágio de blástula) foram expostas (>200 ovos) por 96 horas à diferentes concentrações dos xenobióticos e avaliadas até o décimo dia. Foram analisados os desfechos embrioletalidade, teratogenicidade (indução de malformações) e retardo da eclosão e calculadas as Concentrações Letais 50 por cento (CL50 24h, 96h e 10 dias), as Concentrações Inibitórias 50 por cento (CI50–10 dias) e as Concentrações Efetivas 50 por cento (CE50–10 dias). Os resultados (M) para acetona, etanol e DMSO foram como se segue: Embrioletalidade (CL50 e (IC95 por cento)) 24h: 0,23 (0,22-0,25), 0,53 (0,45-0,56), 1,58 (1,50-1,67); 96h: 0,20 (0,19-0,21), 0,31 (0,29-0,32), 0,20 (0,19-0,21); 10 dias:0,17 (0,16-0,18), 0,26 (0,25-0,27), 0,15 (0,14-0,16). Retardo de Eclosão(CI50 e (IC95 por cento), 10 dias): 0,18 (0,18-0,19), 0,25 (0,21-0,30), 0,09 (0,07-0,11). A CE50 (IC95 por cento)-10 dias para indução de malformações foi 0,17 (0,16-0,18) M. Os valores obtidos (M) para a AS, CP, CdCl2, COL e H2O2 foram: a) CL50 e IC95 por cento-96h: (1,87 e 1,76-1,98)x10-4; (1,71 e 1,70-1,81)x10-2; (2,58 e 2,41-2,77)x10-6; (5,19 e 4,91-5,47)x10-6; (5,72 e 5,60-5,86)x10-7; b) CL50 e [IC95 por cento]-10 dias: (1,61 e 1,52-1,70)x10-4; (1,51 e 1,48-1,64)x10-2; (3,69 e 3,59-3,81)x10-7; (2,19 e 2,08-2,31) x10-6; (5,61 e 5,48-5,74)x10-7; c) CE50 e IC95 por cento -10d: (0,96 e 0,88-1,05)x10-4; (1,31 e 1,16-1,48)x10-2; (3,37 e 3,19-3,55)x10-7; (1,09 e 0,99-1,21)x10-6. d) CI50 e IC95 por cento-10d: (1,45 e 1,31-1,61)x10-4; (0,58 e 0,54-0,62)x10-2; (2,64 e 2,50-2,70)x10-7; (0,79 e 0,73-0,86)x10-6; (4,72 e 4,65-4,79)x10-7. Não foi possível determinar a CE50 para o etanol, DMSO e H2O2. Os resultados mostraram que os embriões de B. glabrata foram susceptíveis aos efeitos tóxicos dos xenobióticos testados e sugerem que o ensaio realizado parece constituir método sensível, simples e de baixo custo para avaliar a embriotoxicidade de poluentes aquáticos.