Microbial Source Tracking Using 16S rRNA Amplicon Sequencing Identifies Evidence of Widespread Contamination from Young Children's Feces in an Urban Slum of Nairobi, Kenya

Child exposure to fecal contamination remains common in low- and middle-income countries after sanitation interventions. Unsafe disposal of children's feces may contribute to this continued exposure, but its relative importance to domestic fecal contamination is not well understood. To address this gap, we interviewed and collected environmental samples (drinking water, caregiver hands, child hands, surfaces, soil, open drainage ditches, standing water, streams) from 40 households in Kibera, an urban slum in Nairobi, Kenya. To track young children's feces (<3 years old) separately from other human-associated fecal sources, we validated distance-based and Bayesian (SourceTracker) microbial source tracking methods using amplicon-based sequencing of the 16S rRNA gene. Contamination by young children's feces could be identified and distinguished separately from older child/adult feces with high sensitivity and specificity in water and soil. Among environmental samples, young children's feces were almost always identified as the dominant source of human fecal contamination inside households (hands, surfaces) whereas older children/adult feces were often identified as the dominant source outside households (standing water, streams, soil). Markers for young children's feces were also detected in standing water and streams, and markers for both fecal sources were equally likely to be dominant in open ditches. These results establish motivation for sanitation interventions that directly address child feces management.