Molecular Techniques in Ecohealth Research Toolkit: Facilitating Estimation of Aggregate Gastroenteritis Burden in an Irrigated Periurban Landscape

Abstract

Assessment of microbial hazards associated with certain environmental matrices, livelihood strategies, and food handling practices are constrained by time-consuming conventional microbiological techniques that lead to health risk assessments of narrow geographic or time scope, often targeting very few pathogens. Health risk assessment based on one or few indicator organisms underestimates true disease burden due a number of coexisting causative pathogens. Here, we employed molecular techniques in a survey of Cryptosporidium parvum, Giardia lamblia, Campylobacter jejuni, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., Shigella spp., Vibrio cholera, and Rotavirus A densities in canal water with respect to seasonality and spatial distribution of point-nonpoint pollution sources. Three irrigational canals stretching across nearly a 150-km(2) periurban landscape, traditionally used for agricultural irrigation but function as vital part of municipal wastewater stabilization in recent years, were investigated. Compiled stochastic data (pathogen concentration, susceptible populations) and literature-obtained deterministic data (pathogen dose-response model parameter values) were used in estimating waterborne gastroenteritis burden. Exposure scenarios include swimming or fishing, consuming canal water-irrigated vegetables, and ingesting or inhaling water aerosols while working in canal water-irrigated fields. Estimated annual gastroenteritis burden due individual pathogens among the sampling points was -10.6log(10) to -2.2log(10) DALYs. Aggregated annual gastroenteritis burden due all the target pathogens per sampling point was -3.1log(10) to -1.9log(10) DALYs, far exceeding WHO acceptable limit of -6.0log(10) DALYs. The present approach will facilitate the comprehensive collection of surface water microbiological baseline data and setting of benchmarks for interventions aimed at reducing microbial hazards in similar landscapes worldwide.

Molecular Analysis for Screening Human Bacterial Pathogens in Municipal Wastewater Treatment and Reuse

“…Routinely used culture-based E. coli detection methods are inadequate in differentiating between pathogenic and nonpathogenic strains of E. coli . Thus, some researchers have used real-time nested PCR to quantify pathogenic E. coli O157:H7 . Nevertheless, the development of 16S rRNA DGGE as a fingerprinting tool in the current study will be helpful in analyzing microdiversity directly from genomic DNA extracted from water samples, bypassing pure culture isolation.…”

“…Thus, some researchers have used real-time nested PCR to quantify pathogenic E. coli O157:H7. 34 Nevertheless, the development of 16S rRNA DGGE as a fingerprinting tool in the current study will be helpful in analyzing microdiversity directly from genomic DNA extracted from water samples, bypassing pure culture isolation. Furthermore, subsequent sequencing of DGGE bands (band length of ∼550 bp) can reveal the identity of the strains.…”

The Use of (Treated) Domestic Wastewater for Irrigation: Current Situation and Future Challenges

“…Current methods for the detection of pathogenic viruses, bacteria, protozoa and helminthes tend to be inaccurate, time-consuming and difficult [24,161]. Culturebased E. coli detection methods, for example, proved to be insufficient in differentiating between pathogenic and nonpathogenic strains [162]. As a result, molecular techniques, including DNA or RNA sequencing, are been used to improve the detection, monitoring and track of specific pathogens in order to understand outbreaks occurrences, forecast transmission dynamics, and detect antibiotic-resistant bacteria, among other objectives [163][164][165].…”