Occurrence, Sources and Virulence Potential of Arcobacter butzleri in Urban Municipal Stormwater Systems.

A. butzleri is an underappreciated emerging global pathogen, despite growing evidence that it is a major contributor of diarrheal illness. Few studies have investigated the occurrence and public health risks that this organism possesses from waterborne exposure routes including through stormwater use. In this study, we assessed the prevalence, virulence potential, and primary sources of stormwater-isolated A. butzleri in fecally contaminated urban stormwater systems. Based on qPCR, A. butzleri was the most common enteric bacterial pathogen [25%] found in stormwater among a panel of pathogens surveyed, including Shiga-toxin producing Escherichia coli (STEC) [6%], Campylobacter spp. [4%], and Salmonella spp. [<1%]. Concentrations of the bacteria, based on qPCR amplification of the single copy gene hsp60, were as high as 6.2 log10 copies/100 mL, suggesting significant loading of this pathogen in some stormwater systems. Importantly, out of 73 unique stormwater culture isolates, 90% were positive for the putative virulence genes cadF, ciaB, tlyA, cjl349, pldA, and mviN, while 50-75% of isolates also possessed the virulence genes irgA, hecA, and hecB. Occurrence of A. butzleri was most often associated with the human fecal pollution marker HF183 in stormwater samples. These results suggest that A. butzleri may be an important bacterial pathogen in stormwater, warranting further study on the risks it represents to public health during stormwater use.

Application of a microbial and pathogen source tracking toolbox to identify infrastructure problems in stormwater drainage networks: a case study.

Water scarcity and increasing urbanization are forcing municipalities to consider alternative water sources, such as stormwater, to fill in water supply gaps or address hydromodification of receiving urban streams. Mounting evidence suggests that stormwater is often contaminated with human feces, even in stormwater drainage systems separate from sanitary sewers. Pinpointing sources of human contamination in drainage networks is challenging given the diverse sources of fecal pollution that can impact these systems and the non-specificity of traditional fecal indicator bacteria (FIB) for identifying these host sources. As such, we used a toolbox approach that encompassed microbial source tracking (MST), FIB monitoring, and bacterial pathogen monitoring to investigate microbial contamination of stormwater in an urban municipality. We demonstrate that human sewage frequently contaminated stormwater (in >50% of routine samples), based on the presence of the human fecal marker HF183, and often exceeded microbial water quality criteria. Arcobacter butzleri, a pathogen of emerging concern, was also detected in >50% of routine samples, with 75% of these pathogen-positive samples also being positive for the human fecal marker HF183, suggesting human municipal sewage as the likely source for this pathogen. MST and FIB were used to track human fecal pollution in the drainage network to the most likely point source of contamination, for which a sewage cross-connection was identified and confirmed using tracer dyes. These results point to the ubiquitous presence of human sewage in stormwater and also provide municipalities with the tools to identify sources of anthropogenic contamination in storm drainage networks.IMPORTANCEWater scarcity, increased urbanization, and population growth are driving municipalities worldwide to consider stormwater as an alternative water source in urban environments. However, many studies suggest that stormwater is relatively poor in terms of microbial water quality, is frequently contaminated with human sewage, and therefore could represent a potential health risk depending on the type of exposure (e.g., irrigation of community gardens). Traditional monitoring of water quality based on fecal bacteria does not provide any information about the sources of fecal pollution contaminating stormwater (i.e., animals/human feces). Herein, we present a case study that uses fecal bacterial monitoring, microbial source tracking, and bacterial pathogen analysis to identify a cross-connection that contributed to human fecal intrusion into an urban stormwater network. This microbial toolbox approach can be useful for municipalities in identifying infrastructure problems in stormwater drainage networks to reduce risks associated with water reuse.