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Wastewater reuse for agriculture is common worldwide; wastewater treatment, however, is rare in many countries, leading to high potential for exposure to harmful pathogens. Mexico City, one of the largest producers of untreated wastewater for agricultural use worldwide, was the site of key epidemiologic studies conducted in the 1990s. We both reviewed the literature on and conducted a cross-sectional study of diarrheal risk and wastewater contamination to provide an updated assessment of health risks and to inform an upcoming update of the 2006 WHO guidelines on wastewater reuse. We surveyed communities in the Mezquital Valley that use wastewater for irrigation and communities that use well water to compare the prevalence of self-reported diarrheal disease in children under five years old. Wastewater, well water, household environmental samples, and stool samples were collected and analyzed. Communities exposed to wastewater had a higher one-week prevalence of diarrhea (10%) compared to unexposed communities (5%). This association remained in an adjusted modified Poisson regression model (PR = 2.31, 95% CI 1.00, 5.31), but not when limited to households engaged in agriculture. Water quality indicators document differences between irrigation water from the two community groups. These results are in agreement with 25 population studies identified by our review that were conducted since or not included in the 2006 WHO guidelines and show consistent negative impacts of wastewater exposure on health. While overall diarrheal prevalence has declined when compared to studies conducted over 25 years ago in the same region, the association of diarrheal disease and wastewater exposure has remained and possibly increased. With rising urbanization worldwide, attention to these risks and wastewater treatment is becoming increasingly important. [ABSTRACT FROM AUTHOR]