Objectives: To estimate the association between improved water and sanitation access and handwashing of mothers living in a peri-urban community of Lima, Peru. Methods: We observed 27 mothers directly, before and after installation of private, piped water and sewerage connections in the street just outside their housing plots, and measured changes in the proportion of faecal-hand contamination and hand-to-mouth transmission events with handwashing. Results: After provision of water and sewerage connections, mothers were approximately two times more likely to be observed washing their hands within a minute of defecation, compared with when they relied on shared, external water sources and non-piped excreta disposal (RR = 2.14, 95% CI = 0.99-4.62). With piped water and sewerage available at housing plots, handwashing with or without soap occurred within a minute after 48% (10/21) of defecation events and within 15 min prior to 8% (11/136) of handling food events. Conclusions: Handwashing increased following installation of private, piped water and sewerage connections, but its practice remained infrequent, particularly before food-related events. Infrastructural interventions should be coupled with efforts to promote hygiene and ensure access to water and soap at multiple on-plot locations convenient to mothers. © 2014 John Wiley & Sons Ltd.