BACKGROUND: Sanitation aims to sequester human feces and prevent exposure to fecal pathogens. More than 2.4 billion people worldwide lack access to improved sanitation facilities and almost one billion practice open defecation. We undertook systematic reviews and meta-analyses to compile the most recent evidence on the impact of sanitation on diarrhea, soil-transmitted helminth (STH) infections, trachoma, schistosomiasis, and nutritional status assessed using anthropometry. METHODS AND FINDINGS: We updated previously published reviews by following their search strategy and eligibility criteria. We searched from the previous review's end date to December 31, 2015. We conducted meta-analyses to estimate pooled measures of effect using random-effects models and conducted subgroup analyses to assess impact of different levels of sanitation services and to explore sources of heterogeneity. We assessed risk of bias and quality of the evidence from intervention studies using the Liverpool Quality Appraisal Tool (LQAT) and Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) approach, respectively. A total of 171 studies met the review's inclusion criteria, including 64 studies not included in the previous reviews. Overall, the evidence suggests that sanitation is protective against diarrhea, active trachoma, some STH infections, schistosomiasis, and height-for-age, with no protective effect for other anthropometric outcomes. The evidence was generally of poor quality, heterogeneity was high, and GRADE scores ranged from very low to high. CONCLUSIONS: This review confirms positive impacts of sanitation on aspects of health. Evidence gaps remain and point to the need for research that rigorously describes sanitation implementation and type of sanitation interventions.