Water, Sanitation and Hygiene (WASH) and environmental risk factors for soil-transmitted helminth intensity of infection in Timor-Leste, using real time PCR

Background: No investigations have been undertaken of risk factors for intensity of soil-transmitted helminth (STH) infection in Timor-Leste. This study provides the first analysis of risk factors for intensity of STH infection, as determined by quantitative PCR (qPCR), examining a broad range of water, sanitation and hygiene (WASH) and environmental factors, among communities in Manufahi District, Timor-Leste. Methods: A baseline cross-sectional survey of 18 communities was undertaken as part of a cluster randomised controlled trial, with additional identically-collected data from six other communities. qPCR was used to assess STH infection from stool samples, and questionnaires administered to collect WASH, demographic, and socioeconomic data. Environmental information was obtained from open-access sources and linked to infection outcomes. Mixed-effects multinomial logistic regression was undertaken to assess risk factors for intensity of Necator americanus and Ascaris infection. Results: 2152 participants provided stool and questionnaire information for this analysis. In adjusted models incorporating WASH, demographic and environmental variables, environmental variables were generally associated with infection intensity for both N. americanus and Ascaris spp. Precipitation (in centimetres) was associated with increased risk of moderate-intensity (adjusted relative risk [ARR] 6.1; 95% confidence interval [CI] 1.9–19.3) and heavy-intensity (ARR 6.6; 95% CI 3.1–14.1) N. americanus infection, as was sandy-loam soil around households (moderate-intensity ARR 2.1; 95% CI 1.0–4.3; heavy-intensity ARR 2.7; 95% CI 1.6–4.5; compared to no infection). For Ascaris, alkaline soil around the household was associated with reduced risk of moderate-intensity infection (ARR 0.21; 95% CI 0.09–0.51), and heavy-intensity infection (ARR 0.04; 95% CI 0.01–0.25). Few WASH risk factors were significant. Conclusion: In this high-prevalence setting, strong risk associations with environmental factors indicate that anthelmintic treatment alone will be insufficient to interrupt STH transmission, as conditions are favourable for ongoing environmental transmission. Integrated STH control strategies should be explored as a priority. Author summary: We present a detailed analysis of WASH, environmental and demographic factors associated with intensity of STH infection in Manufahi District, Timor-Leste, using qPCR. Investigation of risk factors for intensity of STH infection is rarely undertaken, and prior analyses have used microscopic-based eggs per gram of faeces (epg) measures, which are of lower diagnostic accuracy than qPCR. Additionally, few analyses have investigated combined WASH and environmental risk factors in association with STH. This is important due to the extensive potential interrelatedness of environmental, social, behavioural and host factors in any given setting influencing STH survival and transmission. This analysis uses categorical intensity of infection variables for Necator americanus and Ascaris spp., and advanced statistical modelling to adjust for multinomial intensity outcomes, dependency of observations, effects of poverty, and confounding from other measured variables. As such, this analysis provides a comprehensive assessment of risk factors for STH in Manufahi District, Timor-Leste. This is of importance for development of policy and programmatic decisions; risk factors need to be considered not only for their clinical and statistical significance, but more broadly in terms of what may represent modifiable pathways for STH transmission.