Identification of markers of depression and neurotoxicity in pesticide exposed agriculture workers.

Earlier, we reported that chronic exposure to pesticides causes a reduction in the acetylcholinesterase activity and hematological and biochemical alterations in agriculture workers. In continuation with that, the present study aimed to investigate the pesticide-induced neurochemical imbalance and its association with behavior alterations in agricultural workers. A significant increase in depressive symptoms, assessed by the Beck Depression Inventory-II was observed in pesticide exposed workers as compared to the unexposed. A decrease in the level of dopamine in plasma and levels of dopamine, 3,4-dihydroxyphenylacetic acid, homovanillic acids, norepinephrine, serotonin, and hydroxyindoleacetic acid in urine was also observed. An increase in the levels of MAO-A and MAO-B has also been observed in these individuals. The decreased levels of neurotransmitters in the blood and urine have been linked with increased levels of MAO and pesticide residues in plasma and urine. Furthermore, these changes were associated with a higher incidence of depression in agricultural workers.

Salt-assisted acetonitrile extraction and HPLC-QTOF-MS/MS detection for residues of multiple classes of pesticides in human serum samples.

Detecting pesticide residues in human serum is a challenging process. In this study we developed and validated a method for the extraction and recovery of residues of multiple classes of pesticides from serum using one reagent. Salt-assisted acetonitrile extraction and high-performance liquid chromatography with quadrupole time of flight tandem mass spectrometry were used to quantitate 34 pesticides classified in nine groups of chemicals in human serum samples, which are frequently detected in food. The recoveries for 33 of analyzed pesticides ranged from 86 to 112% with relative standard deviations below 15%. The limits of quantitation and linearity of 31 of the pesticides were 1 µg/L and >0.990, respectively. The lower limit of quantitation has been reported in the literature particularly for multi-classes pesticide mixtures in human serum. The salt-acetonitrile reagent was allowed to achieve good recoveries and detection limits, which could be attributed to salt altering the solvent polarity, preferentially collecting the organic phase in the solution, and promoting the extraction. The developed method was applied for two organophosphate pesticide metabolites, diethylphosphate and 3,5,6-trichloro-2-pyridinol, in serum from rats that were fed a nonlethal quantity of chlorpyrifos. The concentrations of these two were 252.18 ± 15.47 and 0.63 ± 0.23 µg/L, respectively.

Determination of insecticide residues and their adverse effects on blood profile of occupationally exposed individuals.

Insecticides, essential for crop protection measures, leave behind several toxic residues that can result in a series of human health disorders. Therefore, this study was planned for the determination of residues and adverse effects of insecticides in blood samples of sprayers, pesticide-industry workers and controls by using blood parameters of these individuals as biomarkers. Optimized analytical methods using GC-MS and HPLC for the simultaneous detection of 22 currently used insecticides were adopted. Eight of twenty-seven (22.22%) sprayers' blood samples were found positive for five different insecticides. Eleven of twenty-seven (40.74%) pesticide-industry workers were found positive for eight different insecticides. The blood samples of both the exposed groups, sprayers and industry workers had significantly (P < 0.001; Mann-Whitney U-tests) low hemoglobin-Hb concentrations (12.17 ±â€¯2.13 and 12.22 ±â€¯2.37 g/dl respectively) than the average value of the control group with 14.23 ±â€¯2.37 g/dl. The erythrocyte sedimentation rates (ESRs) in sprayers and insecticide industry workers (28.78 ±â€¯20.72 and 28.17 ±â€¯25.14 mm/1st h respectively) were greater significantly (P < 0.001; Mann-Whitney U test) than the control blood samples (9.53 ±â€¯3.34 mm/1st h). These results indicate that the exposed individuals have experienced significant hemotoxic effects during insecticide exposure. The study also predicts the risk to exposed individuals in developing countries like Pakistan and demands realization of safety measures to prevent such dangerous effects of pesticide exposures.

Pesticide genotoxicity in cotton picking women in Pakistan evaluated using comet assay.

To control agricultural pests and meet the increasing food demands, pesticides use has been increased substantially over time. Although pesticides are relatively specific to their targets, they can affect non-target organisms and are hazardous for the population around the application areas particularly to the individuals engaged in different types of agricultural activities. This situation is worse in developing and under-developed countries where personal protective equipment is merely used and regulatory guidelines are hardly practiced. In the present study, DNA damage in women exposed to pesticides while picking cotton with bare hands was assessed using single cell gel electrophoresis assay or comet assay. The presence of pesticides in blood serum of exposed individuals was also analyzed using high-performance liquid chromatography. Blood samples were collected from 138 (69 exposed and 69 control) randomly selected females from a major cotton growing area (Bahawalpur District) of the Punjab province of Pakistan. DNA damage, as determined by the mean comet tail length, was significantly higher (p < 0.001) in the exposed group compared to the unexposed. A positive correlation of DNA damage with age and exposure time was also observed. Residues of three pesticides, cyhalothrin, endosulfan, and deltamethrin found significantly higher (p < 0.001) in the serum samples of the exposed group compared to the unexposed. It was observed that the groups with higher mean comet tail length also had a higher concentration of pesticides in their serum samples indicating a positive association of DNA damage and pesticide exposure. The present study suggests that exposure to pesticides leads to DNA damage.

Background biomonitoring of residue levels of 137 pesticides in the blood plasma of the general population in Beijing.

Due to the widespread use of pesticides, human exposure to pesticides is possible and can potentially cause adverse impacts on public health. We measured 137 pesticides including organophosphorus, organochlorine, pyrethroid and carbamate pesticides together with various herbicides in 100 human blood samples collected from the general population in Beijing. The samples were analysed by triple quadrupole tandem gas chromatography-mass spectrometry. In total, 24 organochlorine pesticides, 5 pyrethroid pesticides and 6 organophosphorus pesticides were detected. The detection rates of HCB, α-HCH, ß-HCH, γ-HCH, p,p'-DDE and quintozene were 99, 96, 74, 72, 96 and 95%, respectively. No statistically significant gender difference in the blood concentrations of the pesticides was found. Consistent with the trend of the increasing ß-HCH, p,p'-DDE and quintozene concentrations with age, a strong positive correlation between the age and concentrations of ß-BHC, p,p'-DDE and quintozene was observed.

[Simultaneous determination of 21 organophosphrous pesticides in human serum by QuEChERS-gas chromatography-mass spectrometry].

Objective: A method for the simultaneous determination of organophosphrous pesticides inhuman serum was developed. Methods: Using C18 and PSA as absorbent, 21 organophosphrous pesticides including methamidophos, dichlorvos, acephate, Omethoate, demeton and etc. in human serum were determined by combing QuEChERS with GC-MS, organophosphrous pesticides were qualitatedsimultaneously by retention time and characteristic ions. Their contents were determined by internal standard method of quantitative ions. Results: The linear range was from0to 0.60µg/ml with a good correlation coefficients (r ≥ 0.9961) . The LODs of this methodranged from 0.0009 to 0.0111 µg/ml. The average recoveries of3 organophosphrous pesticides (methamidophos、dichlorvos and acephate) ranged from 71.8%~85.3%, The average recoveries of other 18 organophosphrous pesticidesranged from 82.9%~102.9%. The relative standard deviations were0.8%~7.0%. Conclusion: The convenient operation and versatility of this method are suitable forthe fast screening and accurate detection of 21 organophosphrous pesticides in human serum.

Simultaneous determination of multiclass pesticide residues in human plasma using a mini QuEChERS method.

Blood is one of the most assessable matrices for the determination of pesticide residue exposure in humans. Effective sample preparation/cleanup of biological samples is very important in the development of a sensitive, reproducible, and robust method. In the present study, a simple, cost-effective, and rapid gas chromatography-tandem mass spectrometry method has been developed and validated for simultaneous analysis of 31 multiclass (organophosphates, organochlorines, and synthetic pyrethroids) pesticide residues in human plasma by means of a mini QuEChERS (quick, easy, cheap, effective, rugged, and safe) method. We have adopted a modified version of the QuEChERS method, which is primarily used for pesticide residue analysis in food commodities. The QuEChERS method was optimized by use of different extraction solvents and different amounts and combinations of salts and sorbents (primary-secondary amines and C18) for the dispersive solid-phase extraction step. The results show that a combination of ethyl acetate with 2% acetic acid, magnesium sulfate (0.4 g), and solid-phase extraction for sample cleanup with primary-secondary amines (50 mg) per 1-mL volume of plasma is the most suitable for generating acceptable results with high recoveries for all multiclass pesticides from human plasma. The mean recovery ranged from 74% to 109% for all the analytes. The limit of quantification and limit of detection of the method ranged from 0.12 to 13.53 ng mL-1 and from 0.04 to 4.10 ng mL-1 respectively. The intraday precision and the interday precision of the method were 6% or less and 11% or less respectively. This method would be useful for the analysis of a wide range of pesticides of interest in a small volume of clinical and/or forensic samples to support biomonitoring and toxicological applications. Graphical Abstract Pesticide residues analysis in human plasma using mini QuEChERS method.

Tea consumption in pregnancy as a predictor of pesticide exposure and adverse birth outcomes: The MIREC Study.

INTRODUCTION: Pesticide residues in tea may contribute to exposure during pregnancy; however, the impact on maternal and infant health is not well understood. The aim of this study was to determine whether tea intake in the first trimester was associated with elevated concentrations of various pesticides in maternal blood or urine. Further, we examined the relationship between tea consumption and adverse birth outcomes. METHODS: Data from the Maternal-Infant Research on Environmental Chemicals (MIREC) Study, a pan-Canada pregnancy cohort, were used. All singleton, live births (n=1898) with available biomarkers were included in the analyses. Descriptive statistics were used to characterize the population. The geometric means (GM) of organochlorine (OC) pesticide constituents or metabolites in maternal plasma (lipid adjusted) and organophosphate (OP) pesticide metabolites (adjusted for specific gravity) in maternal urine were calculated for participants who drank regular, green or herbal tea in the first trimester and for those who did not. Differences between groups were examined using chi-square or t-tests. Associations between frequency of drinking tea and adverse birth outcomes were examined using logistic regression (preterm birth and small-for-gestational-age) or generalized linear models (birthweight decile and head circumference). RESULTS: The GM of the OC pesticide constituent trans-nonachlor was 2.74 mg/g lipid, and for metabolites oxychlordane and p,p'-DDE this was 1.94 ng/g lipid and 55.8 ng/g lipid, respectively. OP pesticide metabolite concentrations adjusted for specific gravity, were dimethylphosphate (GM: 3.19 µg/L), dimethylthiophosphate (GM: 3.29 µg/L), dimethyldithiophosphate (GM: 0.48 µg/L), diethlphosphate (GM: 2.46), and diethylthiophosphate (GM: 0.67 µg/L). There was no significant difference in mean concentrations for these OC or OP pesticide constituents or metabolites between tea drinkers - of any type - and non-tea drinkers. Further, no association was found between tea intake and adverse birth outcomes. CONCLUSIONS: Pesticide concentrations did not differ by tea intake. Further, tea intake in the first trimester was not associated with adverse birth outcomes. In this study population, there was no evidence for concern about tea intake being a source of the OP or OC pesticide metabolites measured or adversely affecting birth outcomes; however, tea intake was lower than national Canadian data for women of reproductive age.

Hematological and hepatic alterations in Brazilian population heavily exposed to organochlorine pesticides.

The aim of this study was to investigate the frequency of hematological and hepatic alterations and possible association with serum levels of beta-hexachlorocyclohexane (beta-HCH), p,p'-DDE, and hexachlorobenzene (HCB) among residents in an area heavily contaminated with organochlorine (OC) pesticides. A cross-sectional study was conducted in 415 male and 432 female residents aged >14 years. Serum samples were collected and analyzed for OC pesticides concentrations and biochemical parameters. Frequencies of hematological and hepatic alterations were calculated for each gender. Association between beta-HCH, p,p'-DDE (1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene), and HCB levels and presence of alterations was determined by logistic regression stratified by gender and controlling for confounders. Highest frequencies were observed for eosinophilia (23% men and 18% women), low hemoglobin (12% men and 15% women), and low erythrocyte count (12% men). High levels of bilirubin, glutamic-oxaloacetic transaminase (GOT), and glutamic-pyruvic transaminase (GPT) were observed, respectively, in 10, 11, and 12% of men and <10% of women. Gamma-glutamyl transferase (GGT) was elevated in 26 and 25% of males and females, respectively. Multivariate analysis revealed associations between eosinophilia and beta-HCH in men (OR = 1.06, 95%CI = 1.01-1.12) and women (OR = 1.05, 96%CI = 0.99-1.11), p,p'-DDE in men (OR = 1.03, 95%CI = 0.99-1.06) and women (OR = 1.02, 95%CI = 0.99-1.06), and HCB in women (OR = 1.54, 95%IC = 0.85-4.45). Beta-HCH was found to be associated with increased risk of elevated bilirubin in females (OR = 1.18, 95%CI = 1.07-1.29) and males (OR = 4.21, 95%CI = 1.87-9.47 for fourth vs. first quintile). Thus, OC pesticides may exert adverse effects on hematopoietic tissue and liver in populations chronically exposed to high levels of these compounds.

Monitoring of pesticide residues in human blood from Punjab, India.

In the present study, the current levels of pesticide residues were studied in the human populace of Punjab state. A total of 111 human blood samples were analyzed by gas chromatography and pesticide residues were detected in 35 % of the blood sample(s). Residues of alpha-hexachlorocyclohexane (α-HCH), beta-hexachlorocyclohexane (ß-HCH), p,p'-dichlorodiphenyldichloroethane (p,p' DDD), p,p' dichlorodiphenyldichloroethylene (p,p' DDE), p,p' dichlorodiphenyldichloroethane (p,p' DDT), ß-endosulfan, monocrotophos, profenophos and phosalone were found in human blood samples with mean levels of 1.11, 5.89, 0.51, 3.88, 0.39, 34.90, 0.79, 0.39 and 6.76 ng ml(-1), respectively, with ß-endosulfan as a leading pesticide residue. A paradigm shift in the pattern of the pesticide usage was observed with a shift from organochlorine pesticides to organophosphates.

A review of methods for the analysis of orphan and difficult pesticides: glyphosate, glufosinate, quaternary ammonium and phenoxy acid herbicides, and dithiocarbamate and phthalimide fungicides.

This article reviews the chromatography/MS methodologies for analysis of pesticide residues of orphan and difficult chemical classes in a variety of sample matrixes including water, urine, blood, and food. The review focuses on pesticide classes that are not commonly included in multiresidue analysis methods such as highly polar or ionic herbicides including glyphosate, glufosinate, quaternary ammonium, and phenoxy acid herbicides, and some of their major degradation or metabolite products. In addition, dithiocarbamate and phthalimide fungicides, which are thermally unstable and have stability issues in some solvents or sample matrixes, are also examined due to their special needs in residue analysis.

Organochlorine pesticide residues in biological compartments of healthy mothers.

The present study was conducted to detect placental and breast milk (BM) transfer of organochlorine pesticides (OCPs) between biological compartments in healthy lactating mothers. The study explores the effect of parity, breast-feeding and urban/rural residence on body burden of OCP residues. The study included 38 healthy participants submitted to cesarean delivery. Sociodemographic data and specimens of maternal and umbilical sera, adipose tissue (Adp T) and BM were collected. Specimens were analyzed to detect OCP residues. The lindane in BM, o,p'-dichlorodiphenyldichloroethane (DDD) in maternal serum (MS) and total dichlorodiphenyltrichloroethane (DDT) and o,p'-dichlorodiphenyldichloroethylene in umbilical serum were the only detected residues in significantly higher frequencies and/or means in the primigravidae than multigravidae. There was a high risk of detecting o,p'-DDD (odds ratio = 8.3) in umbilical serum of the mothers with o,p'-DDD detected in the MS. Total DDT was detected in about 65% of specimens of BM, MS and Adp T and in about 40% of the umbilical serum specimens. There was only a significant positive correlation between total DDT residues in the BM and Adp T. Total DDT residues in umbilical serum and Adp T were significantly higher in the urban than in the rural mothers. The detection of some OCPs in maternal biological compartments suggests their potential placental and BM transfer to her child during pregnancy and lactation, respectively. Furthermore, it might reflect the persistence or recent use of these pesticides in the environment.

[Determination of 15 kinds of pesticides in blood by gas chromatography-mass spectrometry].

OBJECTIVE: To develop the method of gas chromatography-mass spectrometry (GC-MS) for simultaneous determination of 15 herbicides in blood. METHODS: 2ml of blood in vitro were sampled, concentrated and extracted with dichloromethane, reconstant with methanol agents of Gulonic acid lactone solution, and detected by GC-MS. RESULTS: Experimental results show that diazinon, atrazine, prometryn, methyl parathion, butachlor, bifenthrin at 4-80 microg/L, phorate, malathion, 2,4-D butyl ester, chlordane, fenpropathrin at 10-200 microg/L, alpha-endosulfan, beta-endosulfan, cyhalothrin at 20-400 microg/L, dimethoate at 40-800 microg/L, with good linear response. The correlation coefficient (r2) were between 0.998-1.000, respectively. The recovery of all analysts averaged between 56%-128% in blood samples. The detection limits of all compounds between 0.05 and 1.00 microg/L. The lower limit of quantification between 0.20 and 3.001 microg/L. CONCLUSION: The methods is apply to detect the content of analysts in blood samples.

Organochlorine pesticide residues in human blood samples collected from Haryana, India and the changing pattern.

Blood samples were collected during August 1992 and August 2002 from various hospitals of Haryana state and analyzed for the presence of HCH and DDT residues and the change in concentration of pesticide residues was calculated in terms of % reduction. The study revealed that the mean residue levels of total HCH in human blood samples have declined by 87.6% while those of total DDT have decreased by 98.9% during a gap of 10 years. The obtained results reveal that during 1992 p,p'-DDT was the major component with the mean value of 6.125 mg/L followed by p,p'-DDE, γ-HCH, α-HCH and ß-HCH while in 2002, ß-HCH and p,p'-DDE were comparable with mean value of 0.053 and 0.052 mg/L, respectively followed by p,p'-DDT, α-HCH and p,p'-DDD.

Organochlorine pesticide residues in blood samples of agriculture and sheep wool workers in Bangalore (rural), India.

To describe exposure level of organochlorine pesticides (OCP) among workers occupationally engaged in agriculture and sheep wool associated jobs, the present study was carried out in rural neighborhood of Bangalore city, India. Thirty participants were interviewed and obtained informed consent before blood sample collection. The maximum concentrations of OCP were detected in blood samples of agriculture workers than sheep wool workers. Among the metabolites of HCH and DDT, lindane (γ-HCH) and p,p'-DDE were the most contributed to the total OCP. There were no differences in pesticide residues found between sex and work groups. It was observed that about 30% of samples exceeded the tolerance limits of 10 µg/L prescribed for HCH under the prevention of food adulteration act. Therefore, the present study recommends continuous monitoring with larger sample size.

Bio-monitoring of persistent organochlorines in human milk and blood samples from sub-Himalayan region of India.

In the present study, concentrations of organochlorine pesticide residues viz. Dichlorodiphenyltrichloroethane and its metabolites (DDTs) and Hexachlorocyclohexane isomers (HCHs) in human breast milk and human blood samples, collected from several high altitude regions of Garhwal Himalaya in Uttarakhand, India viz. Devprayag, Chamoli, Uttarkashi, Joshimath, Bhatwari and Gangnani (altitude ranging from 472 to 1,982 m above sea level) were determined. Mean concentrations of HCH and DDT in human milk samples ranged from 4.53 to 34.32 mg/kg and 6.09 to 12.98 mg/kg, respectively. While the human blood showed mean values ranging from 6.64 to 281.7 µg/L and 12.37 to 104.10 µg/L for HCH and DDT, respectively. The study showed much higher concentrations of organochlorine residue contamination in the Garhwal region as compared to other parts of India. Risk assessments for infants were also calculated and were found within WHO limits.

[Method establishment for the determination of 8 organophosphorus pesticides in serum using LC/MS/MS].

OBJECTIVE: To establish the method to determine the presence and concentration of 8 organophosphors pesticides in serum by LC/MS/MS. METHOD: The serum samples were extracted by ethyl acetateether and then separated on C18 5 microm (2.1 mm x 150 mm) column with pH = 3.5 acid-ammonium acetate buffer solution, methanol--water as mobile phase to mixed gradient elution. Electro-spray ionization source (ESI) was applied and operated in positive ion mode, and multiple reaction monitoring(MRM) mode was adopted. RESULT: Within concentration ranging from 0-1000 ng/ml the correlation coefficient for each compound was greater than 0.990, recoveries were between 95%-129%, RSD < 7.9%. The RSD of intra-day and inter-day were less 12% (n = 6). CONCLUSION: The procedure of preparing serum samples is sample and the result of this method is efficient, accurate and specific.

Organophosphate pesticide exposure among farm women and children: Status of micronutrients, acetylcholinesterase activity, and oxidative stress.

Nutritional status plays a major role in determining the possible adverse health outcomes due to pesticide toxicity. The objective of the present study was to assess the organophosphorus pesticide residue levels among farm women (FW) (24-45years) and farm children (FC) (9-12 and 13-15years) belonging to the Rangareddy district (Telangana, India) along with their micronutrient status, inhibition of acetylcholinesterase activity, and oxidative stress levels. Residues of Chlorpyrifos, Diazinon, Malathion, Monocrotophos and Phosalone were found in the serum samples of FW and FC along with significantly low levels of vitamins and minerals. Inhibition of AChE activity was observed in FW and FC and altered oxidative stress parameters among FW. Correlation studies have found significant associations between the pesticide residues, micronutrients and antioxidant enzymes. The study suggests an association between pesticide exposure coupled with micronutrient deficiency, induced AChE inhibition, and oxidative stress.

Incidence of organochlorine pesticides and the health condition of nestling ospreys (Pandion haliaetus) at Laguna San Ignacio, a pristine area of Baja California Sur, Mexico.

We identified and quantified organochlorine (OC) pesticide residues in the plasma of 28 osprey (Pandion haliaetus) nestlings from a dense population in Laguna San Ignacio, a pristine area of Baja California Sur, Mexico, during the 2001 breeding season. Sixteen OC pesticides were identified and quantified. α-, ß-, δ- and γ-hexachlorocyclohexane, heptaclor, heptachlor epoxide, endosulfan I and II, endosulfan-sulfate, p,p'-DDE, p,p'-DDD, aldrin, dieldrin, endrin, endrin aldehyde, and endrin ketone were the OCs found in the plasma of nestlings, ranging from 0.002 to 6.856 pg/µl (parts per billion). No differences were found in the concentration of pesticides between genders (P > 0.05). In our work, the concentrations detected in the plasma were lower than those reported to be a threat for the species and that affect the survival and reproduction of birds. The presence of OC pesticides in the remote Laguna San Ignacio osprey population is an indication of the ubiquitous nature of these contaminants. OCs are apparently able to travel long distances from their source to the study area. A significant relationship between hemoglobin and mean corpuscular hemoglobin concentrations and OC concentrations were found suggesting that a potential effect on the health of chicks may exist in this osprey population caused by the OC, e.g. anemia. The total proteins were positively correlated with α-BHC, endosulfan I, and p,p'-DDD. It has been suggested that OC also affects competitive interactions and population status over the long term in vertebrate species, and our results could be used as reference information for comparison with other more exposed osprey populations.

Pesticide distribution in an agricultural environment in Argentina.

An assessment of the off-site migration of pesticides from agricultural activity into the environment in the Neuquen River Valley was performed. The aim of this study was to evaluate the distribution of pesticides in several compartments of a small agricultural sub-catchment. Soil, surface water, shallow groundwater and drift deposition were analyzed for pesticide residues. Results showed the presence of some pesticide residues in soil, surface water and shallow groundwater compartments. The highest detection frequencies in water (surface and subsurface) were found for azinphos-methyl and chlorpyrifos (>70%). In terms of concentration, the highest levels were observed in shallow groundwater for azinphos methyl (22.5 µg/L) and carbaryl (45.7 µg/L). In the soil, even before the application period had started, accumulation of residues was present. These residues increased during the period studied. Spray drift during pesticide application was found to be a significant pathway for the migration of pesticide residues in surface water, while leaching and preferential flows were the main transport routes contributing to subsurface contamination.