Antimicrobial resistance (AMR) is a major threat to global health. Understanding the emergence, evolution, and transmission of individual antibiotic resistance genes (ARGs) is essential to develop sustainable strategies combatting this threat. Here, we use metagenomic sequencing to analyse ARGs in 757 sewage samples from 243 cities in 101 countries, collected from 2016 to 2019. We find regional patterns in resistomes, and these differ between subsets corresponding to drug classes and are partly driven by taxonomic variation. The genetic environments of 49 common ARGs are highly diverse, with most common ARGs carried by multiple distinct genomic contexts globally and sometimes on plasmids. Analysis of flanking sequence revealed ARG-specific patterns of dispersal limitation and global transmission. Our data furthermore suggest certain geographies are more prone to transmission events and should receive additional attention.

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This study examines the patterns, trends, and factors associated with functional community water points in rural Ethiopia and identifies potential areas of improvement in terms of practitioner response to functionality and functionality monitoring. It was part of an integrated WaSH and nutrition program implemented by UNICEF Ethiopia and the Government of Ethiopia. Cross-sectional surveys were conducted to collect WaSH-related data in communities and WaSH committees from four community-based nutrition (CBN) program groupings in Ethiopia. In all areas, CBN was implemented, but only in less than half of the areas, a WaSH intervention was implemented. Seventy-three representative kebeles, comprising 30 intervention and 43 control communities, were surveyed. Two structured surveys were conducted. The ‘community survey’ addressed community water points and their functionality and the main areas for improvement needed. The ‘WaSH committee survey’ investigated technical and management aspects of water points and their functionality. Data were analyzed using bivariate regression to identify community characteristics and management practices associated with functionality of water points and explore opportunities to improve water point functionality and monitoring. In the communities, 65% of water points were functional. Eighty percent of communities had a WaSH committee. The WaSH committee members reported that the most used water point types were protected dug wells and boreholes, and that 80% of their water points were functional. India Mark II pumps were more likely to be functional and communities with longer established WaSH committees had higher water point functionality. Communities suggested that the key factors for water point sustainability were improving water quality and water pressure, reducing water collection time, and speeding up repair times. Taking community leaders’ ‘priority lists’ into consideration offers sustainable opportunities for demand-driven, adaptive and targeted design and implementation of rural water supply programs, which, if they include the grassroots level as key informants and actors of change, can succeed. Interventions should integrate the ‘voice’ of the community, the WaSH committees, and other stakeholders and thereby facilitate transdisciplinary approaches at different stages of program management (planning, monitoring, and evaluation). This would help closing the knowledge to action gap and improve policy, programming, practice, and service delivery.

Recreational, reclaimed and drinking source waters worldwide are under increasing anthropogenic pressure, and often contain waterborne enteric bacterial, protozoan, and viral pathogens originating from non-point source fecal contamination. Recently, the capsid integrity (ci)-qPCR, utilizing the azo-dyes propidium monoazide (PMA) or ethidium monoazide (EMA), has been shown to reduce false-positive signals under laboratory conditions as well as in food safety applications, thus improving the qPCR estimation of virions of public health significance. The compatibility of two widely used human adenovirus (HAdV) qPCR protocols was evaluated with the addition of a PMA/EMA pretreatment using a range of spiked and environmental samples. Stock suspensions of HAdV were inactivated using heat, UV, and chlorine before being quantified by cell culture, qPCR, and ci-qPCR. Apparent inactivation of virions was detected for heat and chlorine treated HAdV while there was no significant difference between ci-qPCR and qPCR protocols after disinfection by UV. In a follow-up comparative analysis under more complex matrix conditions, 51 surface and 24 wastewater samples pre/post UV treatment were assessed for enteric waterborne HAdV to evaluate the ability of ci-qPCR to reduce the number of false-positive results when compared to conventional qPCR and cell culture. Azo-dye pretreatment of non-UV inactivated samples was shown to improve the ability of molecular HAdV quantification by reducing signals from virions with an accessible genome, thereby increasing the relevance of qPCR results for public health purposes, particularly suited to resource-limited low and middle-income settings.