Monitoring of diverse enteric pathogens across environmental and host reservoirs with TaqMan array cards and standard qPCR: a methodological comparison study.

BACKGROUND: Multiple bacteria, viruses, protists, and helminths cause enteric infections that greatly impact human health and wellbeing. These enteropathogens are transmited via several pathways through human, animal, and environmental reservoirs. Individual qPCR assays have been extensively used to detect enteropathogens within these types of samples, whereas the TaqMan array card (TAC), which allows simultaneous detection of multiple enteropathogens, has only previously been validated in human clinical samples. METHODS: In this methodological comparison study, we compared the performance of a custom 48-singleplex TAC relative to standard qPCR. We established the sensitivity and specificity of each method for the detection of eight enteric targets, by using spiked samples with varying levels of PCR inhibition. We then tested the prevalence and abundance of pathogens in wastewater from Melbourne (Australia), and human, animal, and environmental samples from informal settlements in Suva, Fiji using both TAC and qPCR. FINDINGS: Both methods exhibited similarly h specificity (TAC 100%, qPCR 94%), sensitivity (TAC 92%, qPCR 100%), and quantitation accuracy (TAC 91%, qPCR 99%) in non-inhibited sample matrices with spiked gene fragments. PCR inhibitors substantially affected detection via TAC, though this issue was alleviated by ten-fold sample dilution. Among samples from informal settlements, the two techniques performed similarly for detection (89% agreement) and quantitation (R2 0·82) for the eight enteropathogen targets. The TAC additionally included 38 other enteric targets, enabling detection of diverse faecal pathogens and extensive environmental contamination that would be prohibitively labour intensive to assay by standard qPCR. INTERPRETATION: The two techniques produced similar results across diverse sample types, with qPCR prioritising greater sensitivity and quantitation accuracy, and TAC trading small reductions in these for a cost-effective larger enteropathogen panel enabling a greater number of enteric pathogens to be analysed concurrently, which is beneficial given the abundance and variety of enteric pathogens in environments such as urban informal settlements. The ability to monitor multiple enteric pathogens across diverse reservoirs could allow better resolution of pathogen exposure pathways, and the design and monitoring of interventions to reduce pathogen load. FUNDING: Wellcome Trust Our Planet, Our Health programme.

Broad approaches to cholera control in Asia: Water, sanitation and handwashing.

Cholera has been eliminated as a public health problem in high-income countries that have implemented sanitation system separating the community's fecal waste from their drinking water and food supply. These expensive, highly-engineered systems, first developed in London over 150 years ago, have not reached low-income high-risk communities across Asia. Barriers to their implementation in communities at highest risk for cholera include the high capital and operating costs for this technological approach, limited capacity and perverse incentives of local governments, and a decreasing availability of water. Interim solutions including household level water treatment, constructing latrines and handwashing promotion have only marginally reduced the risk of cholera and other fecally transmitted diseases. Increased research to develop and policy flexibility to implement a new generation of solutions that are designed specifically to address the physical, financial and political constraints of low-income communities offers the best prospect for reducing the burden of cholera across Asia.

Nitrogen composition in urban runoff--implications for stormwater management.

A study was conducted to characterise the composition of nitrogen in urban stormwater in Melbourne, Australia, during baseflows and storm events, and to compare the results with international data. Nitrogen in Melbourne stormwater was predominantly dissolved (approximately 80%), with ammonia the least-abundant form (approximately 11%). Concentrations of nitrogen species did not vary significantly between baseflow and storms, although the proportion of nitrogen in particulate form was higher during storm events (p = 0.04). Whilst the composition of nitrogen in Melbourne was broadly consistent with international data, the level of dissolved inorganic nitrogen was higher in Melbourne (mu = 48% during baseflows and 49% during storms) than in the international literature (mu = 29%). Limitations in the international dataset precluded comparison of total dissolved nitrogen. The results have implications for stormwater management. Whilst nitrogen species concentrations are variable, they are not strongly related to flow conditions, so treatment systems must be designed to cope with stochastic inflow concentrations at all times. To optimise their performance, stormwater treatments should be designed to improve dissolved nitrogen removal. Further research is needed to improve the ability of treatment systems to achieve this aim.