BackgroundPesticides may impact respiratory health, yet evidence of their impact on pediatric asthma morbidity is limited, particularly among urban children. ObjectiveTo characterize pesticide biomarker concentrations and evaluate their associations with pediatric asthma morbidity among predominantly low-income, Black children in Baltimore City, USA. MethodsWe measured urinary concentrations of 10 biomarkers for pyrethroid insecticides (cyfluthrin:4F-3PBA, permethrin:3PBA), organophosphate insecticides (chlorpyrifos:TCPY, malathion:MDA, parathion:PNP, diazinon:IMPY), and herbicides (glyphosate:AMPA, GPS; 2,4-dicholorphenoxyacetic acid:2,4-D; 2,4,5-tricholorphenoxyacetic acid:2,4,5-T) among 148 children (5–17 years) with established asthma. Urine samples and asthma morbidity measures (asthma symptoms, healthcare utilization, lung function and inflammation) were collected every three months for a year. Generalized estimating equations were used to examine associations between pesticide biomarker concentrations and asthma morbidity measures, controlling for age, sex, race, caregiver education, season, and environmental tobacco smoke. In sensitivity analyses, we assessed the robustness of our results after accounting for environmental co-exposures. ResultsFrequently detected (≥90% detection) pesticide biomarker concentrations (IMPY, 3PBA, PNP, TCPY, AMPA, GPS) varied considerably within children over the follow-up period (intraclass correlation coefficients: 0.1–0.2). Consistent positive significant associations were observed between the chlorpyrifos biomarker, TCPY, and asthma symptoms. Urinary concentrations of TCPY were associated with increased odds of coughing, wheezing, or chest tightness (adjusted Odds Ratio, aOR, TCPY:1.60, 95% Confidence Interval, CI:1.17–2.18). Urinary concentrations of TCPY were also associated with maximal symptom days (aOR:1.38, CI:1.02–1.86), exercise-related symptoms (aOR:1.63, CI:1.09–2.44), and hospitalizations for asthma (aOR:2.84, CI:1.08–7.43). We did not observe consistent evidence of associations between the pesticide exposures assessed and lung function or inflammation measures. ConclusionAmong predominantly Black children with asthma, we found evidence that chlorpyrifos is associated with asthma morbidity. Further research is needed to assess the contribution of pesticide exposures to pediatric respiratory health and characterize exposure sources among vulnerable populations to inform targeted interventions against potentially harmful pesticide exposures.