Legionella detection in wastewater treatment plants with increased risk for Legionella growth and emission.

Legionnaires' disease (LD) is a severe pneumonia mainly caused by the bacterium Legionella pneumophila. Although many environmental sources of LD have been described, the sources of the majority of non-outbreak LD cases have not been identified. In several outbreaks in the Netherlands, wastewater treatment plants (WWTPs) were identified as the most likely source of infection. In this study, four criteria for Legionella growth and emission to air and surface waters were selected based on the literature and a risk matrix was drafted. An inventory was made of all WWTPs and their characteristics in the Netherlands. The risk matrix was applied to identify WWTPs at risk for Legionella growth and emission. Wastewater was collected at WWTPs with moderate to high risk for Legionella growth and emission. In 18% of the sampled WWTPs, Legionella spp. was detected using culture methods. The presented risk matrix can be used to assess the risks of Legionella growth and emission for WWTPs and support surveillance by prioritizing WWTPs. When Legionella is detected in the wastewater, it is recommended to take action to prevent emission to air or discharge on surface waters and, if possible, reduce the Legionella concentration.

The effective design of sampling campaigns for emerging chemical and microbial contaminants in drinking water and its resources based on literature mining.

As well as known contaminants, surface waters also contain an unknown variety of chemical and microbial contaminants which can pose a risk to humans if surface water is used for the production of drinking water. To protect human health proactively, and in a cost-efficient way, water authorities and drinking water companies need early warning systems. This study aimed to (1) assess the effectiveness of screening the scientific literature to direct sampling campaigns for early warning purposes, and (2) detect new aquatic contaminants of concern to public health in the Netherlands. By screening the scientific literature, six example contaminants (3 chemical and 3 microbial) were selected as potential aquatic contaminants of concern to the quality of Dutch drinking water. Stakeholders from the Dutch water sector and various information sources were consulted to identify the potential sources of these contaminants. Based on these potential contamination sources, two sampling sequences were set up from contamination sources (municipal and industrial wastewater treatment plants), via surface water used for the production of drinking water to treated drinking water. The chemical contaminants, mycophenolic acid, tetrabutylphosphonium compounds and Hexafluoropropylene Oxide Trimer Acid, were detected in low concentrations and were thus not expected to pose a risk to Dutch drinking water. Colistin resistant Escherichia coli was detected for the first time in Dutch wastewater not influenced by hospital wastewater, indicating circulation of bacteria resistant to this last-resort antibiotic in the open Dutch population. Four out of six contaminants were thus detected in surface or wastewater samples, which showed that screening the scientific literature to direct sampling campaigns for both microbial and chemical contaminants is effective for early warning purposes.

Evaluation of water quality guidelines for public swimming ponds.

Swimming ponds are artificial ecosystems for bathing in which people imitate the conditions of natural waters. Swimming in natural water may pose health risks if the water quality is microbiologically poor. Swimming ponds are small water bodies that may be used by relatively large groups of people, moreover, the water is not disinfected, e.g. by using chlorine. The draft new swimming pool legislation in the Netherlands includes water quality requirements for swimming ponds. This study focused on the examination and evaluation of the new microbiological water quality requirements, including Escherichia coli, intestinal enterococci, Pseudomonas aeruginosa and Staphylococcus aureus, in thirteen public swimming pools. In eight of thirteen swimming ponds the water quality met the requirements for fecal indicators; 93-95% of the samples met the requirement for E. coli (≤100/100 ml) and intestinal enterococci (≤50/100 ml). The requirement for P. aeruginosa (≤10/100 ml) was met in eleven of thirteen swimming ponds (99% of the samples). In 68% of the samples the requirement for S. aureus (<1/100 ml) was met. A linear mixed effect analysis showed that E. coli and intestinal enterococci concentrations were significantly dependent on the log10 of turbidity. P. aeruginosa concentrations were significantly dependent on water temperature. 31-45% of the variation between swimming ponds was explained by considering 'pond' as a random effect in the analysis. The monitoring of microbiological parameters in swimming pond water needs selective analytical methods, such as those used in this study, due to large numbers of background bacteria. The draft new Dutch swimming pool legislation provides proper guidance to ensure the microbiological safety of swimming pond water; it would benefit from inclusion of turbidity as an extra parameter. S. aureus is a relevant parameter for non-fecal shedding, although scientific literature does not provide evidence for a norm value based on a dose-response relation for exposure to S. aureus in water.

Linking water quality monitoring and climate-resilient water safety planning in two urban drinking water utilities in Ethiopia.

Unsafe drinking water is a recognized health threat in Ethiopia, and climate change, rapid population growth, urbanization and agricultural practices put intense pressure on availability and quality of water. Climate change-related health problems due to floods and waterborne diseases are increasing. With increasing insight into impacts of climate change and urbanization on water availability and quality and of required adaptations, a shift towards climate-resilient water safety planning was introduced into an Ethiopian strategy and guidance document to guarantee safe drinking water. Climate-resilient water safety planning was implemented in the urban water supplies of Addis Ababa and Adama, providing drinking water to 5 million and 500,000 people, respectively. Based on the risks identified with climate-resilient water safety planning, water quality monitoring can be optimized by prioritizing parameters and events which pose a higher risk for contaminating the drinking water. Water quality monitoring was improved at both drinking water utilities and at the Public Health Institute to provide relevant data used as input for climate-resilient water safety planning. By continuously linking water quality monitoring and climate-resilient water safety planning, utilization of information was optimized, and both approaches benefit from linking these activities.

Multidrug-Resistant and Extended Spectrum Beta-Lactamase-Producing Escherichia coli in Dutch Surface Water and Wastewater.

OBJECTIVE: The goal of the current study was to gain insight into the prevalence and concentrations of antimicrobial resistant (AMR) Escherichia coli in Dutch surface water, and to explore the role of wastewater as AMR contamination source. METHODS: The prevalence of AMR E. coli was determined in 113 surface water samples obtained from 30 different water bodies, and in 33 wastewater samples obtained at five health care institutions (HCIs), seven municipal wastewater treatment plants (mWWTPs), and an airport WWTP. Overall, 846 surface water and 313 wastewater E. coli isolates were analysed with respect to susceptibility to eight antimicrobials (representing seven different classes): ampicillin, cefotaxime, tetracycline, ciprofloxacin, streptomycin, sulfamethoxazole, trimethoprim, and chloramphenicol. RESULTS: Among surface water isolates, 26% were resistant to at least one class of antimicrobials, and 11% were multidrug-resistant (MDR). In wastewater, the proportions of AMR/MDR E. coli were 76%/62% at HCIs, 69%/19% at the airport WWTP, and 37%/27% and 31%/20% in mWWTP influents and effluents, respectively. Median concentrations of MDR E. coli were 2.2×10(2), 4.0×10(4), 1.8×10(7), and 4.1×10(7) cfu/l in surface water, WWTP effluents, WWTP influents and HCI wastewater, respectively. The different resistance types occurred with similar frequencies among E. coli from surface water and E. coli from municipal wastewater. By contrast, among E. coli from HCI wastewater, resistance to cefotaxime and resistance to ciprofloxacin were significantly overrepresented compared to E. coli from municipal wastewater and surface water. Most cefotaxime-resistant E. coliisolates produced ESBL. In two of the mWWTP, ESBL-producing variants were detected that were identical with respect to phylogenetic group, sequence type, AMR-profile, and ESBL-genotype to variants from HCI wastewater discharged onto the same sewer and sampled on the same day (A1/ST23/CTX-M-1, B23/ST131/CTX-M-15, D2/ST405/CTX-M-15). CONCLUSION: In conclusion, our data show that MDR E. coli are omnipresent in Dutch surface water, and indicate that municipal wastewater significantly contributes to this occurrence.

Prevalence and characterization of ESBL- and AmpC-producing Enterobacteriaceae on retail vegetables.

In total 1216 vegetables obtained from Dutch stores during 2012 and 2013 were analysed to determine the prevalence of 3rd-generation cephalosporin (3GC) resistant bacteria on soil-grown fresh produce possibly consumed raw. Vegetables grown conventionally and organically, from Dutch as well as foreign origin were compared. Included were the following vegetable types; blanched celery (n=192), bunched carrots (n=190), butterhead lettuce (n=137), chicory (n=96), endive (n=188), iceberg lettuce (n=193) and radish (n=120). Overall, 3GC-resistant Enterobacteriaceae were detected on 5.2% of vegetables. Based on primary habitat and mechanism of 3GC-resistance, these bacteria could be divided into four groups: ESBL-producing faecal species (Escherichia coli, Enterobacter spp.), AmpC-producing faecal species (Citrobacter freundii, Enterobacter spp.), ESBL-producing environmental species (Pantoea spp., Rahnella aquatilis, Serratia fonticola), and AmpC-producing environmental species (Cedecca spp., Hafnia alvei, Pantoea spp., Serratia plymuthica), which were detected on 0.8%, 1.2%, 2.6% and 0.4% of the vegetables analysed, respectively. Contamination with faecal 3GC-resistant bacteria was most frequently observed in root and bulb vegetables (average prevalence 4.4%), and less frequently in stem vegetables (prevalence 1.6%) and leafy greens (average prevalence 0.6%). In Dutch stores, only four of the included vegetable types (blanched celery, bunched carrots, endive, iceberg lettuce) were available in all four possible variants: Dutch/conventional, Dutch/organic, foreign/conventional, foreign/organic. With respect to these vegetable types, no statistically significant difference was observed in prevalence of 3GC-resistant Enterobacteriaceae between country of origin or cultivation type (5.2%, 5.7%, 5.7% and 3.3%, respectively). Vegetables consumed raw may be a source of dissemination of 3GC-resistant Enterobacteriaceae and their resistance genes to humans. The magnitude of the associated public health risk presumably depends on the types of bacteria that are ingested, i.e., faecal or environmental species, and may therefore be higher for root and bulb vegetables compared to leafy greens.

Extended spectrum ß-lactamase- and constitutively AmpC-producing Enterobacteriaceae on fresh produce and in the agricultural environment.

The attribution of fresh produce to the overall community-associated exposure of humans to ESBL- or AmpC-producing bacteria is currently unknown. To address this issue, the prevalence of ESBL- and AmpC-producing Enterobacteriaceae on fresh produce produced in the Netherlands was determined. Seven vegetable types that are consumed raw were selected: blanched celery, bunched carrots, chicory, endive, iceberg lettuce, mushrooms, and radish. The vegetables were mostly obtained from supermarkets. To determine whether the agricultural environment is the source of ESBL-producing Enterobacteriaceae on fresh produce, iceberg lettuce was also obtained directly from three farms, in conjunction with soil and irrigation water. ESBL-producing Enterobacteriaceae isolated from vegetables and environment were all environmental species: Rahnella aquatilis (n = 119), Serratia fonticola (n = 45) and Pantoea agglomerans (n = 1). ESBL genes of R. aquatilis and S. fonticola were identified as blaRAHN-1 and blaRAHN-2 and blaFONA-1, blaFONA-2, blaFONA-3/6 and blaFONA-5, respectively. For R. aquatilis and S. fonticola, different prevalence numbers were observed using different isolation methods, which could at least partially be explained by an inverse correlation between the level of cefotaxime resistance of these species and incubation temperature. R. aquatilis was isolated from 0 to 46% of soil samples and 11 to 83% of vegetable samples, and S. fonticola from 2 to 60% of soil samples and 0 to 1.3% of vegetable samples. Third generation cephalosporin-resistant faecal Enterobacteriaceae were isolated from 2.7%, 1.3% and 1.1% of supermarket vegetables, iceberg lettuce from farms, and agricultural soil respectively. Faecal Enterobacteriaceae were all identified as Citrobacter and Enterobacter species and, with the exception of one Citrobacter koseri strain, all had phenotypes indicative of constitutive AmpC production. Comparison of fresh produce and its agricultural environment indicates that the Enterobacteriaceae population on fresh produce reflects that of the soil it is grown in. Public health risks associated with exposure to ESBL- and AmpC-producing bacteria through consumption of uncooked fresh produce are diverse. They range from occasional ingestion of 3GC-resistant opportunistic pathogens which may result in difficult-to-treat infections, to frequent ingestion of relatively harmless ESBL-producing environmental bacteria that may therewith constitute a continuously replenished intestinal reservoir facilitating dissemination of ESBL genes to (opportunistic) pathogens.