Multi-state study of Enterobacteriaceae harboring extended-spectrum beta-lactamase and carbapenemase genes in U.S. drinking water.

Community-associated acquisition of extended-spectrum beta-lactamase- (ESBL) and carbapenemase-producing Enterobacteriaceae has significantly increased in recent years, necessitating greater inquiry into potential exposure routes, including food and water sources. In high-income countries, drinking water is often neglected as a possible source of community exposure to antibiotic-resistant organisms. We screened coliform-positive tap water samples (n = 483) from public and private water systems in six states of the United States for blaCTX-M, blaSHV, blaTEM, blaKPC, blaNDM, and blaOXA-48-type genes by multiplex PCR. Positive samples were subcultured to isolate organisms harboring ESBL or carbapenemase genes. Thirty-one samples (6.4%) were positive for blaCTX-M, ESBL-type blaSHV or blaTEM, or blaOXA-48-type carbapenemase genes, including at least one positive sample from each state. ESBL and blaOXA-48-type Enterobacteriaceae isolates included E. coli, Kluyvera, Providencia, Klebsiella, and Citrobacter species. The blaOXA-48-type genes were also found in non-fermenting Gram-negative species, including Shewanella, Pseudomonas and Acinetobacter. Multiple isolates were phenotypically non-susceptible to third-generation cephalosporin or carbapenem antibiotics. These findings suggest that tap water in high income countries could serve as an important source of community exposure to ESBL and carbapenemase genes, and that these genes may be disseminated by non-Enterobacteriaceae that are not detected as part of standard microbiological water quality testing.

A pilot study on the assessment of trace organic contaminants including pharmaceuticals and personal care products from on-site wastewater treatment systems along Skaneateles Lake in New York State, USA.

On-site wastewater treatment systems (OWTSs or septic systems) are designed to treat and dispose effluents on the same property that produces the wastewater. Approximately 25% of the U.S. population is served by such facilities. Nevertheless, studies on the treatment efficiency and discharge of organic contaminants through septic effluents are lacking. This pilot study showed the occurrence of organic contaminants including pharmaceuticals and personal care products (PPCPs), perfluoroalkyl surfactants (PFASs), polybrominated diphenyl ethers (PBDEs), and polychlorinated biphenyls (PCBs) in septic effluents, adjacent lake water samples, household drinking water in homes that use lake water or a well adjacent to the lake as a source of drinking water, and offshore lake water samples. Septic effluent as well as lake and tap water samples were collected from several households with OWTSs around Skaneateles Lake located in central New York. The advanced on-site systems were installed in some households for the purpose of limiting nutrient levels in the effluent to protect the local surface water. Additionally, because many of these systems serve homes with limited land, advanced treatment systems were needed. The median concentrations of ten PPCPs (ranged from 0.45 to 388 ng/L) and eleven PFASs (ranged from 0.20 to 14.6 ng/L) in septic water were significantly higher (p ≤ 0.01) than in lake water samples. The median concentrations of PPCPs and PFASs in lake and tap water samples were not significantly different (p ≥ 0.65). The median concentrations of ∑PBDEs in septic, lake, and tap water samples were 7.47, 3.49, and 2.22 ng/L, respectively, and those for ∑PCBs were 33.1, 29.2, and 28.6 ng/L, respectively. The mass flux of PPCPs (i.e. the mass flow of PPCPs per unit area per unit time) through the disposal of treated septic effluent from textile biofilter and aerobic treatments to the dispersal unit ranged from 12 (carbamazepine) to 66900 µg/m(2)/day (caffeine) whereas that for PFASs ranged from 7.0 (perfluorobutanesulfonate) to 833 µg/m(2)/day (perfluorooctanoic acid). Based on the ratio of measured concentrations and method detection limit, triclocarban, perfluorooctanoic acid, and perfluorooctanesulfonate have the potential to be used as chemical tracers of septic water contamination in Skaneateles Lake. The median concentrations of atenolol, a beta-blocker drug, in septic water were significantly (ρ = 0.86, p = 0.01) correlated with enterococci counts.

Growth of Escherichia coli in model distribution system biofilms exposed to hypochlorous acid or monochloramine.

Bacteria indigenous to water distribution systems were used to grow multispecies biofilms within continuous-flow slide chambers. Six flow chambers were also inoculated with an Escherichia coli isolate obtained from potable water. The effect of disinfectants on bacterial populations was determined after exposure of established biofilms to 1 ppm of hypochlorous acid (ClOH) for 67 min or 4 ppm of monochloramine (NH(2)Cl) for 155 min. To test the ability of bacterial populations to initiate biofilm formation in the presence of disinfectants, we assessed the biofilms after 2 weeks of exposure to residual concentrations of 0.2 ppm of ClOH or 4 ppm of NH(2)Cl. Lastly, to determine the effect of recommended residual concentrations on newly established biofilms, we treated systems with 0.2 ppm of ClOH after 5 days of growth in the absence of disinfectant. Whole-cell in situ hybridizations using fluorescently tagged, 16S rRNA-targeted oligonucleotide probes performed on cryosectioned biofilms permitted the direct observation of metabolically active bacterial populations, including certain phylogenetic groups and species. The results of these studies confirmed the resistance of established bacterial biofilms to treatment with recommended levels of disinfectants. Specifically, Legionella pneumophila, E. coli, and beta and delta proteobacteria were identified within biofilms both before and after treatment. Furthermore, although it was undetected using routine monitoring techniques, the observation of rRNA-containing E. coli within biofilms demonstrated not only survival but also metabolic activity of this organism within the model distribution systems. The persistence of diverse bacterial species within disinfectant-treated biofilms suggests that current testing practices underestimate the risk to immunocompromised individuals of contracting waterborne disease.

Detection, isolation, and molecular subtyping of Escherichia coli O157:H7 and Campylobacter jejuni associated with a large waterborne outbreak.

The largest reported outbreak of waterborne Escherichia coli O157:H7 in the United States occurred in upstate New York following a county fair in August 1999. Culture methods were used to isolate E. coli O157:H7 from specimens from 128 of 775 patients with suspected infections. Campylobacter jejuni was also isolated from stools of 44 persons who developed diarrheal illness after attending this fair. There was one case of a confirmed coinfection with E. coli O157:H7 and C. jejuni. Molecular detection of stx(1) and stx(2) Shiga toxin genes, immunomagnetic separation (IMS), and selective culture enrichment were utilized to detect and isolate E. coli O157:H7 from an unchlorinated well and its distribution points, a dry well, and a nearby septic tank. PCR for stx(1) and stx(2) was shown to provide a useful screen for toxin-producing E. coli O157:H7, and IMS subculture improved recovery. Pulsed-field gel electrophoresis (PFGE) was used to compare patient and environmental E. coli O157:H7 isolates. Among patient isolates, 117 of 128 (91.5%) were type 1 or 1a (three or fewer bands different). Among the water distribution system isolates, 13 of 19 (68%) were type 1 or 1a. Additionally, PFGE of C. jejuni isolates revealed that 29 of 35 (83%) had indistinguishable PFGE patterns. The PFGE results implicated the water distribution system as the main source of the E. coli O157:H7 outbreak. This investigation demonstrates the potential for outbreaks involving more than one pathogen and the importance of analyzing isolates from multiple patients and environmental samples to develop a better understanding of bacterial transmission during an outbreak.