Sex and adrenal hormones in association with insecticide biomarkers among adolescents living in ecuadorian agricultural communities.

BACKGROUND: Organophosphate, pyrethroid, and neonicotinoid insecticides have resulted in adrenal and gonadal hormone disruption in animal and in vitro studies; limited epidemiologic evidence exists in humans. We assessed relationships of urinary insecticide metabolite concentrations with adrenal and gonadal hormones in adolescents living in Ecuadorean agricultural communities. METHODS: In 2016, we examined 522 Ecuadorian adolescents (11-17y, 50.7% female, 22% Indigenous; ESPINA study). We measured urinary insecticide metabolites, blood acetylcholinesterase activity (AChE), and salivary testosterone, dehydroepiandrosterone (DHEA), 17ß-estradiol, and cortisol. We used general linear models to assess linear (ß = % hormone difference per 50% increase of metabolite concentration) and curvilinear relationships (ß2 = hormone difference per unit increase in squared ln-metabolite) between ln-metabolite or AChE and ln-hormone concentrations, stratified by sex, adjusting for anthropometric, demographic, and awakening response variables. Bayesian Kernel Machine Regression was used to assess non-linear associations and interactions. RESULTS: The organophosphate metabolite malathion dicarboxylic acid (MDA) had positive associations with testosterone (ßboys = 5.88% [1.21%, 10.78%], ßgirls = 4.10% [-0.02%, 8.39%]), and cortisol (ßboys = 6.06 [-0.23%, 12.75%]. Para-nitrophenol (organophosphate) had negatively-trending curvilinear associations, with testosterone (ß2boys = -0.17 (-0.33, -0.003), p = 0.04) and DHEA (ß2boys = -0.49 (-0.80, -0.19), p = 0.001) in boys. The neonicotinoid summary score (ßboys = 5.60% [0.14%, 11.36%]) and the neonicotinoid acetamiprid-N-desmethyl (ßboys = 3.90% [1.28%, 6.58%]) were positively associated with 17ß-estradiol, measured in boys only. No associations between the pyrethroid 3-phenoxybenzoic acid and hormones were observed. In girls, bivariate response associations identified interactions of MDA, Para-nitrophenol, and 3,5,6-trichloro-2-pyridinol (organophosphates) with testosterone and DHEA concentrations. In boys, we observed an interaction of MDA and Para-nitrophenol with DHEA. No associations were identified for AChE. CONCLUSIONS: We observed evidence of endocrine disruption for specific organophosphate and neonicotinoid metabolite exposures in adolescents. Urinary organophosphate metabolites were associated with testosterone and DHEA concentrations, with stronger associations in boys than girls. Urinary neonicotinoids were positively associated with 17ß-estradiol. Longitudinal repeat-measures analyses would be beneficial for causal inference.

Urinary Glyphosate, 2,4-D and DEET Biomarkers in Relation to Neurobehavioral Performance in Ecuadorian Adolescents in the ESPINA Cohort.

BACKGROUND: Herbicides are the most used class of pesticides worldwide, and insect repellents are widely used globally. Yet, there is a dearth of studies characterizing the associations between these chemical groups and human neurobehavior. Experimental studies suggest that glyphosate and 2,4-dichlorophenoxyacetic acid (2,4-D) herbicides can affect neurobehavior and the cholinergic and glutamatergic pathways in the brain. We aim to assess whether herbicides and insect repellents are associated with neurobehavioral performance in adolescents. METHODS: We assessed 519 participants (11-17 years of age) living in agricultural communities in Ecuador. We quantified urinary concentrations of glyphosate, 2,4-D, and two N,N-diethyl-meta-toluamide (DEET) insect repellent metabolites [3-(diethylcarbamoyl)benzoic acid (DCBA) and 3-(ethylcarbamoyl)benzoic acid (ECBA)] using isotope-dilution mass spectrometry. We assessed neurobehavioral performance using 9 subtests across 5 domains (attention/inhibitory control, memory/learning, language, visuospatial processing, and social perception). We characterized the associations using generalized estimating equations and multiple imputation for metabolites below detection limits. Models were adjusted for demographic and anthropometric characteristics, urinary creatinine, and sexual maturation. Mediation by salivary cortisol, dehydroepiandrosterone, 17ß-estradiol, and testosterone was assessed using structural equation modeling. RESULTS: The mean of each neurobehavioral domain score was between 7.0 and 8.7 [standard deviation (SD) range: 2.0-2.3]. Glyphosate was detected in 98.3% of participants, 2,4-D in 66.2%, DCBA in 63.3%, and ECBA in 33.4%. 2,4-D was negatively associated with all neurobehavioral domains, but statistically significant associations were observed with attention/inhibition [score difference per 50% higher metabolite concentration (ß)=-0.19 95% confidence interval (CI): -0.31, -0.07], language [ß=-0.12 (95% CI: -0.23, -0.01)], and memory/learning [ß=-0.11 (95% CI: -0.22, 0.01)]. Glyphosate had a statistically significant negative association only with social perception [ß=-0.08 (95% CI: -0.14, -0.01)]. DEET metabolites were not associated with neurobehavioral performance. Mediation by gender and adrenal hormones was not observed. CONCLUSION: This study describes worse neurobehavioral performance associated with herbicide exposures in adolescents, particularly with 2,4-D. Replication of these findings among other pediatric and adult populations is needed. https://doi.org/10.1289/EHP11383.

Concurrent urinary organophosphate metabolites and acetylcholinesterase activity in Ecuadorian adolescents.

BACKGROUND: Organophosphates are insecticides that inhibit the enzymatic activity of acetylcholinesterase (AChE). Because of this, AChE is considered a physiological marker of organophosphate exposure in agricultural settings. However, limited research exists on the associations between urinary organophosphate metabolites and AChE activity in children. METHODS: This study included 526 participants from 2 exams (April and July-October 2016) of ages 12-17 years living in agricultural communities in Ecuador. AChE activity was measured at both examinations, and organophosphate metabolites, including para-nitrophenol (PNP), 3,5,6-trichloro-2-pyridinol (TCPy), and malathion dicarboxylic acid (MDA) were measured in urine collected in July-October. We used generalized estimating equation generalized linear model (GEEGLM), adjusting for hemoglobin, creatinine, and other demographic and anthropometric covariates, to estimate associations of urinary metabolite concentrations with AChE activity (July-October) and AChE% change between April and July-October. RESULTS: The mean (SD) of AChE and AChE% change (April vs July-October) were 3.67 U/mL (0.54) and -2.5% (15.4%), respectively. AChE activity was inversely associated with PNP concentration, whereas AChE% change was inversely associated with PNP and MDA. There was evidence of a threshold: difference was only significant above the 80th percentile of PNP concentration (AChE difference per SD increase of metabolite = -0.12 U/mL [95%CI: 0.20, -0.04]). Likewise, associations with AChE% change were significant only above the 80th percentile of TCPy (AChE % change per SD increase of metabolite = -1.38% [95%CI: 2.43%, -0.32%]) and PNP -2.47% [95%CI: 4.45%, -0.50%]). PNP concentration at ≥80th percentile was associated with elevated ORs for low AChE activity of 2.9 (95% CI: 1.5, 5.7) and for AChE inhibition of ≤ -10% of 3.7 (95% CI: 1.4, 9.8). CONCLUSIONS: Urinary organophosphate metabolites, including PNP, TCPy and MDA, particularly at concentrations above the 80th percentile, were associated with lower AChE activity among adolescents. These findings bring attention to the value of using multiple constructs of pesticide exposure in epidemiologic studies.

Intercepted journeys: Associations between migration and mobility experiences and depressive symptoms among substance using migrants at the Mexico-Guatemala border.

Substance use and depressive psychiatric symptoms have been associated with migration and mobility. The Mexico-Guatemala border is a key transit point for internal, regional, and international migration flows. However, there is limited knowledge of the role of substance use, migration, and mobility on mental health among migrants at this border. Our paper explores the association of migration and mobility patterns with possible major depressive symptoms among migrants at this key geographic region. We recruited 392 substance-using migrants using modified time-location sampling. Crude and adjusted logistic regression models were developed. We found that 12% of the sample had possible major depressive symptoms. After adjusting for relevant covariates, including gender, income, and perceived homelessness, we found that recent rural-urban and short-term migrants had higher odds of possible major depressive symptoms, whereas international migrants had lower odds. Findings of this paper suggest that although migrants experience hardship and uncertainty, they may respond with complex and nuanced forms of coping and planning.

Residential proximity to greenhouse crops and pesticide exposure (via acetylcholinesterase activity) assessed from childhood through adolescence.

BACKGROUND: Off-target drift of pesticides from farms increases the risk of pesticide exposure of people living nearby. Cholinesterase inhibitors (i.e. organophosphates and carbamates) are frequently used in agriculture and inhibit acetylcholinesterase (AChE) activity. Greenhouse agriculture is an important production method, but it is unknown how far pesticide drift from greenhouses can extend and expose people living nearby. METHODS: This study included 1156 observations from 3 exams (2008, Apr, 2016 and Jul-Oct 2016) of 623 children aged 4-to-17 years living in agricultural communities in Ecuador. AChE, a physiological marker of cholinesterase inhibitor exposure, was measured in blood. Geographic positioning of greenhouses and homes were obtained using GPS receivers and satellite imagery. Distances between homes and the nearest greenhouse edge, and areas of greenhouse crops within various buffer zones around homes were calculated. Repeated-measures regression adjusted for hemoglobin and other covariates estimated change in AChE relative to distance from greenhouses. RESULTS: The pooled mean (SD) of AChE activity was 3.58 U/mL (0.60). The median (25th-75th %tile) residential distance to crops was 334 m (123, 648) and crop area within 500 m of homes (non-zero values only) was 18,482 m2 (7115, 61,841). Residential proximity to greenhouse crops was associated with lower AChE activity among children living within 275 m of crops (AChE difference per 100 m of proximity [95% CI] = -0.10 U/mL [-0.20, -0.006]). Lower AChE activity was associated with greater crop area within 500 m of homes (AChE difference per 1000 m2 [95% CI] = -0.026 U/mL [-0.040, -0.012]) and especially within 150 m (-0.037 U/mL [-0.065, -0.007]). CONCLUSIONS: Residential proximity to floricultural greenhouses, especially within 275 m, was associated with lower AChE activity among children, reflecting greater cholinesterase inhibitor exposure from pesticide drift. Analyses of residential proximity and crop areas near homes yielded complementary findings. Mitigation of off-target drift of pesticides from crops onto nearby homes is recommended.

Summary data of home proximity to the nearest greenhouse (floricultural) crops and areas of greenhouse crops around various distances from homes in agricultural settings in Ecuador.

This article presents data of 623 children (1156 observations) between 4 and 17 years of age living in floricultural communities of Ecuador from 3 examination periods (2008, Apr 2016, and Jul-Oct 2016) as part of the study of Secondary Exposure to Pesticides among Children and Adolescents (ESPINA).We present geospatial data of residential distance to the nearest greenhouse crop and areas within various buffer sizes around homes of participants which is data used in the original research article: Suarez-Lopez JR, et al. "Residential proximity to greenhouse crops and pesticide exposure (via acetylcholinesterase activity) assessed from childhood through adolescence". These geospatial variables are related but different constructs of the potential for off-target drift of pesticides from crops onto homes nearby (via acetylcholinesterase activity) Understanding the associations between these distance and crop area variables is important as a growing number of investigations are using these constructs of off-target pesticide drift to characterize their associations with both exposure biomarkers and outcome measures. Geographic positioning of greenhouses and homes were obtained using GPS receivers and satellite imagery. Distances between homes and the nearest greenhouse edge, and areas of greenhouse crops within various buffer zones (0-150 m, 151-300 m, 310-500 m, 501-750 m, and 751-1000 m) around homes were calculated using Geographic Information System software. Beyond the dataset, we present demographic and anthropometric characteristics, and indicators of pesticide exposure of participants across categories of areas of greenhouse crops around homes for buffer sizes of 0-150 m around homes. We also present the distribution of areas of flower crops within various buffer zone sizes around children's homes and the correlation coefficients between household proximity to the nearest treated greenhouse crops and areas of flower crops within various buffer zones within 1000 m of homes.

Associations of acetylcholinesterase activity with depression and anxiety symptoms among adolescents growing up near pesticide spray sites.

BACKGROUND: The cholinergic system has an important role in mood regulation. Cholinesterase inhibitor pesticides (e.g. organophosphates) appear to increase depression and anxiety symptoms in the few existing animal and human studies. Human studies have not described such associations using biomarkers of exposure and studies among children are needed. METHODS: We studied 529 adolescents (ages 11-17y) in agricultural communities in the Ecuadorian Andes (ESPINA study). Acetylcholinesterase (AChE) activity was measured in a finger-stick sample. Anxiety and depression symptoms were assessed using the CDI-2 and MASC-2 (greater scores reflect greater internalizing symptoms). Models adjusted for age, gender, hemoglobin, income among others. RESULTS: The median age was 14.38y and 51% were female. The mean (SD) of the following parameters were: AChE 3.7 U/mL (0.55), depression T-score 53.0 (9.4) and anxiety T-score: 57.6 (9.8). Lower AChE activity (reflecting greater cholinesterase inhibitor exposure) was associated with higher depression symptoms (difference per SD decrease of AChE [ß [95% CI:]]: 1.09 [0.02, 2.16]), was stronger among girls (ß = 1.61) than boys (ß = 0.69), and among younger (<14.38y, ß = 1.61) vs. older children (ß = 0.57). The associations were strongest among girls <14.38y (ß = 3.30 [0.54, 6.05], OR for elevated symptoms per SD decrease in AChE = 2.58 [1.26, 5.27]). No associations were observed with anxiety scores. Analyses of AChE change between 2008 and 2016 concurred with these findings. DISCUSSION: We observed associations between a biomarker of pesticide exposure and children's depression symptoms. Lower AChE activity may create risk for depression in teenagers, particularly among girls during early adolescence.

Blood pressure after a heightened pesticide spray period among children living in agricultural communities in Ecuador.

INTRODUCTION: Agricultural pesticide spray periods increase the pesticide exposure potential of children living nearby and growing evidence indicates that they may affect children's health. We examined the association of time following a heightened agricultural production period, the Mother's Day flower harvest (May), with children's blood pressure (BP). METHODS: We included cross-sectional information of 313 children ages 4-9 years in Ecuadorian agricultural communities (the ESPINA study). Examinations occurred during a period of low flower production, but within 63-100 days (mean = 81.5, SD = 10.9) following the Mother's Day harvest. BP was measured twice using a pediatric sphygmomanometer and BP percentiles appropriate for age, gender and height were calculated. RESULTS: Participants were 51% male, 1.6% hypertensive and 7.7% had elevated BP. The mean (SD) BP percentiles were: systolic: 51.7 (23.9); diastolic: 33.3 (20.3). There was an inverse relationship between of time after the spray season with percentiles of systolic (difference [ß] per 10.9 days after the harvest: -4.3 [95%CI: -6.9, -1.7]) and diastolic BP (ß: -7.5 [-9.6, -5.4]) after adjusting for race, heart rate and BMI-for-age z-score. A curvilinear association with diastolic BP was observed. For every 10.9 days that a child was examined sooner after the harvest, the OR of elevated BP/hypertension doubled (OR: 2.0, 95% CI: 1.3, 3.1). Time after the harvest was positively associated with acetylcholinesterase. CONCLUSIONS: Children examined sooner after a heightened pesticide spray period had higher blood pressure and pesticide exposure markers than children examined later. Further studies with multiple exposure-outcome measures across pesticide spray periods are needed.

Home proximity to flower plantations and higher systolic blood pressure among children.

BACKGROUND: Pesticide drift from agricultural plantations increases the chemical exposure potential of people living nearby. Some studies have described positive associations between pesticide exposures and blood pressure (BP) in adults, whereas limited evidence in children suggests negative associations. This study characterized the association between home proximity to plantations and BP among children living in a flower-growing county in Ecuador. METHODS: We included 310 4-9-year-old children living in Pedro Moncayo County, Ecuador as part of The ESPINA study. We calculated age, gender and height-specific BP z-scores. Geographic coordinates of homes and flower plantations were collected using GPS receivers and satellite imagery. Exposure-outcome associations were analyzed using linear regression. RESULTS: The mean home distance to the nearest flower plantation was 449 m (SD: 347) and the median plantation area within 150 m of participants' homes was 989 m2 (25th-75Th percentile: 492-3164) among those with non-zero values. Children living closer to plantations had lower AChE activity. Systolic BP z-score increased with greater residential proximity to plantations (0.24 SD per 1000 m [95% CI: 0.01, 0.47]) and with greater areas of flower plantations within 150 m of homes (0.03 SD per 1000 m2 [0.00, 0.06]), after adjusting for socio-economic, anthropometric and other factors. Further adjustment for acetylcholinesterase and hemoglobin strengthened these associations. CONCLUSIONS: Proximity of homes to flower plantations and greater plantation areas within 150 m from homes were associated with higher systolic BP, independent of cholinesterase activity. This suggests that non-cholinesterase inhibitor pesticide drift from agricultural plantations may be sufficient to induce physiologic changes on children living nearby.

Potential short-term neurobehavioral alterations in children associated with a peak pesticide spray season: The Mother's Day flower harvest in Ecuador.

BACKGROUND: Exposures to cholinesterase inhibitor pesticides (e.g. organophosphates) have been associated with children's neurobehavioral alterations, including attention deficit and impulsivity. Animal studies have observed transient alterations in neurobehavioral performance in relation to cholinesterase inhibitor pesticide exposures; however, limited evidence exists regarding transient effects in humans. METHODS: We estimated the associations between neurobehavioral performance and time after Mother's Day flower harvest (the end of a heightened pesticide usage period) among 308 4-to 9-year-old children living in floricultural communities in Ecuador in 2008 who participated in the ESPINA study. Children's neurobehavior was examined once (NEPSY-II: 11 subtests covering 5 domains), between 63 and 100days (SD: 10.8days) after Mother's Day harvest (blood acetylcholinesterase activity levels can take 82days to normalize after irreversible inhibition with organophosphates). RESULTS: The mean (SD) neurobehavioral scaled scores across domains ranged from 6.6 (2.4) to 9.9 (3.3); higher values reflect greater performance. Children examined sooner after Mother's Day had lower neurobehavioral scores than children examined later, in the domains of (score difference per 10.8days, 95%CI): Attention/Inhibitory Control (0.38, 0.10-0.65), Visuospatial Processing (0.60, 0.25-0.95) and Sensorimotor (0.43, 0.10-0.77). Scores were higher with longer time post-harvest among girls (vs. boys) in Attention/Inhibitory Control. CONCLUSIONS: Our findings, although cross-sectional, are among the first in non-worker children to suggest that a peak pesticide use period may transiently affect neurobehavioral performance, as children examined sooner after the flower harvest had lower neurobehavioral performance than children examined later. Studies assessing pre- and post-exposure measures are needed.

Acetylcholinesterase activity and neurodevelopment in boys and girls.

BACKGROUND: Organophosphate exposures can affect children's neurodevelopment, possibly due to neurotoxicity induced by acetylcholinesterase (AChE) inhibition, and may affect boys more than girls. We tested the hypothesis that lower AChE activity is associated with lower neurobehavioral development among children living in Ecuadorian floricultural communities. METHODS: In 2008, we examined 307 children (age: 4-9 years; 52% male) and quantified AChE activity and neurodevelopment in 5 domains: attention/executive functioning, language, memory/learning, visuospatial processing, and sensorimotor (NEPSY-II test). Associations were adjusted for demographic and socioeconomic characteristics and height-for-age, flower worker cohabitation, and hemoglobin concentration. RESULTS: Mean ± standard deviation AChE activity was 3.14 ± 0.49 U/mL (similar for both genders). The range of scores among neurodevelopment subtests was 5.9 to 10.7 U (standard deviation: 2.6-4.9 U). Girls had a greater mean attention/executive functioning domain score than boys. In boys only, there were increased odds ratios of low (<9th percentile) neurodevelopment among those in the lowest tertile versus the highest tertile of AChE activity (odds ratios: total neurodevelopment: 5.14 [95% confidence interval (CI): 0.84 to 31.48]; attention/executive functioning domain: 4.55 [95% CI: 1.19 to 17.38], memory/learning domain: 6.03 [95% CI: 1.17 to 31.05]) after adjustment for socioeconomic and demographic factors, height-for-age, and hemoglobin. Within these domains, attention, inhibition and long-term memory subtests were most affected. CONCLUSIONS: Low AChE activity was associated with deficits in neurodevelopment, particularly in attention, inhibition, and memory in boys but not in girls. These critical cognitive skills affect learning and academic performance. Added precautions regarding secondary occupational pesticide exposure would be prudent.

Acetylcholinesterase activity, cohabitation with floricultural workers, and blood pressure in Ecuadorian children.

BACKGROUND: Acetylcholinesterase (AChE) inhibitors are commonly used pesticides that can effect hemodynamic changes through increased cholinergic stimulation. Children of agricultural workers are likely to have paraoccupational exposures to pesticides, but the potential physiological impact of such exposures is unclear. OBJECTIVES: We investigated whether secondary pesticide exposures were associated with blood pressure and heart rate among children living in agricultural Ecuadorian communities. METHODS: This cross-sectional study included 271 children 4-9 years of age [51% cohabited with one or more flower plantation workers (mean duration, 5.2 years)]. Erythrocyte AChE activity was measured using the EQM Test-mate system. Linear regression models were used to estimate associations of systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate with AChE activity, living with flower workers, duration of cohabitation with a flower worker, number of flower workers in the child's home, and number of practices that might increase children's exposure to pesticides. RESULTS: Mean (± SD) AChE activity was 3.14 ± 0.49 U/mL. A 1-U/mL decrease in AChE activity was associated with a 2.86-mmHg decrease in SBP (95% CI: -5.20, -0.53) and a 2.89-mmHg decrease in DBP (95% CI: -5.00, -0.78), after adjustment for potential confounders. Children living with flower workers had lower SBP (-1.72 mmHg; 95% CI: -3.53, 0.08) than other children, and practices that might increase exposure also were associated with lower SBP. No significant associations were found between exposures and heart rate. CONCLUSIONS: Our findings suggest that subclinical secondary exposures to pesticides may affect vascular reactivity in children. Additional research is needed to confirm these findings.

Lower acetylcholinesterase activity among children living with flower plantation workers.

BACKGROUND: Children of workers exposed to pesticides are at risk of secondary pesticide exposure. We evaluated the potential for lower acetylcholinesterase activity in children cohabiting with fresh-cut flower plantation workers, which would be expected from organophosphate and carbamate insecticide exposure. Parental home surveys were performed and acetylcholinesterase activity was measured in 277 children aged 4-9 years in the Secondary Exposure to Pesticides among Infants, Children and Adolescents (ESPINA) study. Participants lived in a rural county in Ecuador with substantial flower plantation activity. RESULTS: Mean acetylcholinesterase activity was 3.14 U/ml, standard deviation (SD) of 0.49. It was lower by 0.09 U/ml (95% confidence interval (CI) -0.19, -0.001) in children of flower workers (57% of participants) than non-flower workers' children, after adjustment for gender, age, height-for-age, hemoglobin concentration, income, pesticide use within household lot, pesticide use by contiguous neighbors, examination date and residence distance to nearest flower plantation. Using a 4 level polychotomous acetylcholinesterase activity dependent variable, flower worker cohabitation (vs. not) had odds ratio 3.39 (95% CI 1.19, 9.64) for being <15th percentile compared to the highest tertile. Children cohabitating for ≥5 years (vs. never) had OR of 4.11 (95% CI: 1.17, 14.38) of AChE activity within <15th percentile compared to the highest tertile. CONCLUSIONS: Cohabitation with a flower worker was related to lower acetylcholinesterase activity in children. This supports the hypothesis that the amount of take-home pesticides from flower workers suffices to decrease acetylcholinesterase activity, with lower activity associated with longer exposure.