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BACKGROUND: Glyphosate is one of the most heavily used pesticides in the world, but little is known about sources of glyphosate exposure in pregnant people living in agricultural regions. OBJECTIVE: Our objective was to evaluate glyphosate exposure during pregnancy in relation to residential proximity to agriculture as well as agricultural spray season. METHODS: We quantified glyphosate concentrations in 453 urine samples collected biweekly from a cohort of 40 pregnant people in southern Idaho from February through December 2021. We estimated each participant’s glyphosate exposure as the geometric mean (GM) of glyphosate concentrations measured in all samples (average 푛= 11 samples/participant), as well as the GM of samples collected during the pesticide “spray season” (defined as those collected 1 May–15 August; average 푛= 5 samples/participant) and the “nonspray season” (defined as those collected before 1 May or after 15 August; average 푛= 6 samples/participant). We defined participants who resided < 0.5 km from an actively cultivated agriculture field to live “near fields” and those residing = 0.5 km from an agricultural field to live “far from fields” (푛= 22 and 18, respectively). RESULTS: Among participants living near fields, urinary glyphosate was detected more frequently and at significantly increased GM concentrations during the spray season in comparison with the nonspray season (81% vs. 55%; 0.228 μg/L vs. 0.150 μg/L, 푝 < 0.001). In contrast, among participants who lived far from fields, neither glyphosate detection frequency nor GMs differed in the spray vs nonspray season (66% vs. 64%; 0.154 μg/L vs. 0.165 μg/L, 푝= 0.45). Concentrations did not differ by residential proximity to fields during the nonspray season (0.154 μg/L vs. 0.165μg/L, for near vs. far, 푝= 0.53). DISCUSSION: Pregnant people living near agriculture fields had significantly increased urinary glyphosate concentrations during the agricultural spray season than during the nonspray season. They also had significantly higher urinary glyphosate concentrations during the spray season than those who lived far from agricultural fields at any time of year, but concentrations did not differ during the nonspray season. These findings suggest that agricultural glyphosate spray is a source of exposure for people living near fields. [ABSTRACT FROM AUTHOR]