A membrane filtration method for the enumeration of Escherichia coli in bathing water and other waters with high levels of background bacteria.

The presence and level of faecal indicator bacteria are important factors in estimating the microbiological quality of surface water and the risk of human infection upon exposure to this water. Until 2014, ISO 9308-1:2000 was available and used to enumerate faecal indicator Escherichia coli in bathing water. In 2014, this ISO was technically revised and replaced by ISO 9308-1:2014. This ISO introduced a less selective method for enumeration of E. coli that allows non-specific growth from waters containing high levels of bacteria, such as surface waters. This implies that currently there is no suitable reference membrane filtration method for the compliance monitoring of official bathing sites for E. coli according to the European Bathing Water Directive. Here, the performance characteristics of three chromogenic culture media, namely Tryptone Bile X-glucuronide (TBX) agar, Chromogenic Coliform Agar (CCA), and CHROMagar E. coli/Coliform (ECC) were investigated at 44 °C for water with varying levels of bacteria according to ISO 13843:2017. Based on performance characteristics, colony counts, and practical usage, TBX appeared the most suitable culture medium for the enumeration of E. coli in bathing water and other waters with high levels of background bacteria, such as surface water in agricultural areas and wastewater discharge points.

High genetic diversity of Vibrio cholerae in the European lake Neusiedler See is associated with intensive recombination in the reed habitat and the long-distance transfer of strains.

Coastal marine Vibrio cholerae populations usually exhibit high genetic diversity. To assess the genetic diversity of abundant V. cholerae non-O1/non-O139 populations in the Central European lake Neusiedler See, we performed a phylogenetic analysis based on recA, toxR, gyrB and pyrH loci sequenced for 472 strains. The strains were isolated from three ecologically different habitats in a lake that is a hot-spot of migrating birds and an important bathing water. We also analyzed 76 environmental and human V. cholerae non-O1/non-O139 isolates from Austria and other European countries and added sequences of seven genome-sequenced strains. Phylogenetic analysis showed that the lake supports a unique endemic diversity of V. cholerae that is particularly rich in the reed stand. Phylogenetic trees revealed that many V. cholerae isolates from European countries were genetically related to the strains present in the lake belonging to statistically supported monophyletic clades. We hypothesize that the observed phenomena can be explained by the high degree of genetic recombination that is particularly intensive in the reed stand, acting along with the long distance transfer of strains most probably via birds and/or humans. Thus, the Neusiedler See may serve as a bioreactor for the appearance of new strains with new (pathogenic) properties.

Prevalence and types of Campylobacter on poultry farms and in their direct environment.

To study whether broiler and layer farms contribute to the environmental Campylobacter load, environmental matrices at or close to farms, and caecal material from chickens, were examined. Similarity between Campylobacter from poultry and environment was tested based on species identification and Multilocus Sequence Typing. Campylobacter prevalence in caecal samples was 97% at layer farms (n = 5), and 93% at broiler farms with Campylobacter-positive flocks (n = 2/3). Campylobacter prevalence in environmental samples was 24% at layer farms, and 29% at broiler farms with Campylobacter-positive flocks. Campylobacter was detected in soil and surface water, not in dust and flies. Campylobacter prevalence in adjacent and remote surface waters was not significantly (P > 0.1) different. Detected species were C. coli (52%), C. jejuni (40%) and C. lari (7%) in layers, and C. jejuni (100%) in broilers. Identical sequence types (STs) were detected in caecal material and soil. A deviating species distribution in surface water adjacent to farms indicated a high background level of environmental Campylobacter. STs from layer farms were completely deviant from surface water STs. The occasional detection of identical STs in broilers, wastewater at broiler farms and surface water in the farm environment suggested a possible contribution of broiler farms to the aquatic environmental Campylobacter load.

A QMRA for the Transmission of ESBL-Producing Escherichia coli and Campylobacter from Poultry Farms to Humans Through Flies.

The public health significance of transmission of ESBL-producing Escherichia coli and Campylobacter from poultry farms to humans through flies was investigated using a worst-case risk model. Human exposure was modeled by the fraction of contaminated flies, the number of specific bacteria per fly, the number of flies leaving the poultry farm, and the number of positive poultry houses in the Netherlands. Simplified risk calculations for transmission through consumption of chicken fillet were used for comparison, in terms of the number of human exposures, the total human exposure, and, for Campylobacter only, the number of human cases of illness. Comparing estimates of the worst-case risk of transmission through flies with estimates of the real risk of chicken fillet consumption, the number of human exposures to ESBL-producing E. coli was higher for chicken fillet as compared with flies, but the total level of exposure was higher for flies. For Campylobacter, risk values were nearly consistently higher for transmission through flies than for chicken fillet consumption. This indicates that the public health risk of transmission of both ESBL-producing E. coli and Campylobacter to humans through flies might be of importance. It justifies further modeling of transmission through flies for which additional data (fly emigration, human exposure) are required. Similar analyses of other environmental transmission routes from poultry farms are suggested to precede further investigations into flies.

Fate of Extended-Spectrum ß-Lactamase-Producing Escherichia coli from Faecal Sources in Surface Water and Probability of Human Exposure through Swimming.

The goal of this study was to determine the fate of ESBL-producing Escherichia coli (ESBL-EC) emitted from faecal sources in surface water, and the probability of human exposure through swimming. Concentrations of ESBL-EC were measured in recreational waters and in source waters, being water in ditches surrounding poultry farms and municipal wastewater. Additionally, the potential of ESBL-EC in source waters to reach recreational waters located downstream of these sources was modeled. Modeled ESBL-EC concentrations in recreational waters appeared to be mostly determined by the concentrations in the source waters and by subsequent dilution in surface water, and less by inactivation. The mean (95%) risk of human exposure to ESBL-EC per person per swimming event, as assessed from measured ESBL-EC concentrations in recreational waters, was 0.16 (0.89) for men, 0.13 (0.72) for women and 0.20 (0.95) for children. Similar exposure risks were estimated for hypothetical recreational waters containing 100- or 1000-times diluted source water, located 10 days water travel time downstream of the sources. Human exposure to ESBL-EC through swimming is likely, if recreational waters are located downstream of poultry farms and municipal wastewater discharge points.

Effect of Climate Change on the Concentration and Associated Risks of Vibrio Spp. in Dutch Recreational Waters.

Currently, the number of reported cases of recreational- water-related Vibrio illness in the Netherlands is low. However, a notable higher incidence of Vibrio infections has been observed in warm summers. In the future, such warm summers are expected to occur more often, resulting in enhanced water temperatures favoring Vibrio growth. Quantitative information on the increase in concentration of Vibrio spp. in recreational water under climate change scenarios is lacking. In this study, data on occurrence of Vibrio spp. at six different bathing sites in the Netherlands (2009-2012) were used to derive an empirical formula to predict the Vibrio concentration as a function of temperature, salinity, and pH. This formula was used to predict the effects of increased temperatures in climate change scenarios on Vibrio concentrations. For Vibrio parahaemolyticus, changes in illness risks associated with the changed concentrations were calculated as well. For an average temperature increase of 3.7 °C, these illness risks were calculated to be two to three times higher than in the current situation. Current illness risks were, varying per location, on average between 10(-4) and 10(-2) per person for an entire summer. In situations where water temperatures reached maximum values, illness risks are estimated to be up to 10(-2) and 10(-1) . If such extreme situations occur more often during future summers, increased numbers of ill bathers or bathing-water-related illness outbreaks may be expected.

Detection of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli on flies at poultry farms.

In the Netherlands, extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli bacteria are highly prevalent in poultry, and chicken meat has been implicated as a source of ESBL-producing E. coli present in the human population. The current study describes the isolation of ESBL-producing E. coli from house flies and blow flies caught at two poultry farms, offering a potential alternative route of transmission of ESBL-producing E. coli from poultry to humans. Overall, 87 flies were analyzed in 19 pools. ESBL-producing E. coli bacteria were detected in two fly pools (10.5%): a pool of three blow flies from a broiler farm and a pool of eight house flies from a laying-hen farm. From each positive fly pool, six isolates were characterized and compared with isolates obtained from manure (n = 53) sampled at both farms and rinse water (n = 10) from the broiler farm. Among six fly isolates from the broiler farm, four different types were detected with respect to phylogenetic group, sequence type (ST), and ESBL genotype: A0/ST3519/SHV-12, A1/ST10/SHV-12, A1/ST58/SHV-12, and B1/ST448/CTX-M-1. These types, as well as six additional types, were also present in manure and/or rinse water at the same farm. At the laying-hen farm, all fly and manure isolates were identical, carrying blaTEM-52 in an A1/ST48 genetic background. The data imply that flies acquire ESBL-producing E. coli at poultry farms, warranting further evaluation of the contribution of flies to dissemination of ESBL-producing E. coli in the community.

Sources and transmission routes of campylobacteriosis: A combined analysis of genome and exposure data.

OBJECTIVES: To determine the contributions of several animal and environmental sources of human campylobacteriosis and identify source-specific risk factors. METHODS: 1417 Campylobacter jejuni/coli isolates from the Netherlands in 2017-2019 were whole-genome sequenced, including isolates from human cases (n = 280), chickens/turkeys (n = 238), laying hens (n = 56), cattle (n = 158), veal calves (n = 49), sheep/goats (n = 111), pigs (n = 110), dogs/cats (n = 100), wild birds (n = 62), and surface water (n = 253). Questionnaire-based exposure data was collected. Source attribution was performed using core-genome multilocus sequence typing. Risk factors were determined on the attribution estimates. RESULTS: Cases were mostly attributed to chickens/turkeys (48.2%), dogs/cats (18.0%), cattle (12.1%), and surface water (8.5%). Of the associations identified, never consuming chicken, as well as frequent chicken consumption, and rarely washing hands after touching raw meat, were risk factors for chicken/turkey-attributable infections. Consuming unpasteurized milk or barbecued beef increased the risk for cattle-attributable infections. Risk factors for infections attributable to environmental sources were open water swimming, contact with dog faeces, and consuming non-chicken/turkey avian meat like game birds. CONCLUSIONS: Poultry and cattle are the main livestock sources of campylobacteriosis, while pets and surface water are important non-livestock sources. Foodborne transmission is only partially consistent with the attributions, as frequency and alternative pathways of exposure are significant.

Pathogenic Vibrio species in dutch shellfish destined for direct human consumption.

Vibrio parahaemolyticus is a common cause of shellfish-related gastroenteritis all over the world. V. parahaemolyticus and Vibrio alginolyticus have previously been detected in water samples from the Oosterschelde, a large inlet on the North Sea, which is used for both recreational purposes and shellfish production. In 2006, oysters (Crassostrea gigas) from a noncommercial oyster bed in the Oosterschelde and oysters bought in Dutch fish shops were tested for the presence of pathogenic Vibrio species; in 2007 and 2008, oysters (C. gigas) and mussels (Mytilus edulis) from Oosterschelde production areas were examined. Total Vibrio numbers were related to water temperatures to study joint patterns. Vibrio was found in oysters and mussels from the production areas, and levels ranged from 6 to 622 most probable number (MPN) per g in oysters and 6 to 62 MPN/g in mussels. Vibrio levels in oysters from fish shops were 231 to >333 MPN/g, whereas levels in noncommercial oysters ranged from 231 to >2,398 MPN/g. About 80% of the isolated strains were V. alginolyticus, and approximately 10% were identified as V. parahaemolyticus. Vibrio counts in shellfish samples increased with increasing water temperature and declined when water temperatures dropped; Vibrio was not detected when water temperatures declined to <13.5 degrees C. Based on the obtained results and the known high V. parahaemolyticus dose (>10(4) cells per serving of oysters) required for infection, it is concluded that the risk of gastrointestinal infections with V. parahaemolyticus through consumption of shellfish from the Oosterschelde production sites is presumably low.

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.

Tracing the animal sources of surface water contamination with Campylobacter jejuni and Campylobacter coli.

Campylobacter jejuni and C. coli, the primary agents of human bacterial gastroenteritis worldwide, are widespread in surface water. Several animal sources contribute to surface water contamination with Campylobacter, but their relative contributions thus far remained unclear. Here, the prevalence, genotype diversity, and potential animal sources of C. jejuni and C. coli strains in surface water in the Netherlands were investigated. It was also assessed whether the contribution of the different animal sources varied according to surface water type (i.e. agricultural water, surface water at discharge points of wastewater treatment plants [WWTPs], and official recreational water), season, and local livestock (poultry, pig, ruminant) density. For each surface water type, 30 locations spread over six areas with either high or low density of poultry, ruminants, or pigs, were sampled once every season in 2018-2019. Campylobacter prevalence was highest in agricultural waters (77%), and in autumn and winter (74%), and lowest in recreational waters (46%) and in summer (54%). In total, 76 C. jejuni and 177 C. coli water isolates were whole-genome sequenced. Most C. coli water isolates (78.5%) belonged to hitherto unidentified clones when using the seven-locus sequence type (ST) scheme, while only 11.8% of the C. jejuni isolates had unidentified STs. The origin of these isolates, as defined by core-genome multi-locus sequence typing (cgMLST), was inferred by comparison with Campylobacter strain collections from meat-producing poultry, laying hens, adult cattle, veal calves, small ruminants, pigs, and wild birds. Water isolates were mainly attributed to wild birds (C. jejuni: 60.0%; C. coli: 93.7%) and meat-producing poultry (C. jejuni: 18.9%; C. coli: 5.6%). Wild bird contribution was high among isolates from recreational waters and WWTP discharge points, and in areas with low poultry (C. coli) or high ruminant (C. jejuni) densities. The contribution of meat-producing poultry was high in areas with high density of poultry, springtime, agricultural waters and WWTP discharge points. While wild birds and poultry were the main contributors to Campylobacter contamination in surface water, their contribution differed significantly by water type, season, and local poultry and ruminant densities.

Potential transmission pathways of clinically relevant fungi in indoor swimming pool facilities.

Possible transmission pathways of fungi in indoor swimming pool facilities were assessed through fungal counting in different areas of the facilities and typing of the collected fungal isolates. Air, water and surface samples were collected from seven different indoor swimming pool facilities. Fungal species were identified based on their internal transcribed spacer (ITS) sequences. Maximum fungal concentrations of 6.2 CFU/cm2, 1.39 CFU/100 mL, and 202 CFU/m³ were found on surfaces, in water and air, respectively. In total, 458 isolates were obtained, belonging to 111 fungal species, of which 50 species were clinically relevant. Phialophora oxyspora (13.3%) and Trichosporon dohaense (5.0%) were the most frequently isolated species and were merely detected on floors, as were the dermatophytes, Trichophyton interdigitale and T. rubrum. Penicillium spp. and Aspergillus spp. were the dominant fungi in water and air. No typical patterns of fungal concentrations along the preferential pathways of pool visitors were observed, however, sites where pool visitors converge while moving from one room (e.g. dressing room) to another (e.g. shower room) and walking barefoot displayed the highest fungal concentrations thus posing the highest risk of contamination. The dispersal of fungi on floors is most likely facilitated by the pool visitors and cleaning tools. Clinically relevant fungi, including the ones rarely identified in nature, were widely detected on floors, in water and in air, as well as on cleaning tools and flexibeams. Preventive measures such as cleaning should minimize the prevalence of clinically relevant fungi in swimming pool facilities since these potentially pose health risks to those vulnerable for infections.

Norovirus Outbreak Associated with Swimming in a Recreational Lake Not Influenced by External Human Fecal Sources in The Netherlands, August 2012.

Swimming in fecally contaminated recreational water may lead to gastrointestinal illness. A recreational water-associated outbreak of norovirus (NoV) infections affecting at least 100 people in The Netherlands occurred in August 2012. Questionnaire responses from patients indicated swimming in recreational lake Zeumeren as the most likely cause of illness. Most patients visited the lake during the weekend of 18⁻19 August, during which the weather was exceptionally warm (maximum temperatures 32⁻33 °C), and visitor numbers elevated. Patients, mostly children, became ill with gastroenteritis 1⁻6 days (median 2 days) after exposure. Four stool samples from patients were NoV GI positive. Subsurface sandy soil from one of the beaches where most patients swam was NoV GI positive; the water sample was negative. The epidemiological curve and the timeline of investigation based on reported symptoms demonstrate the difficulty in discovering the source in recreational water outbreaks. A NoV outbreak in a recreational lake that is not subjected to external fecal contamination sources shows the need for active communication about human shedding of viruses during and after diarrheal episodes and the advice to refrain from swimming, even a few weeks after the symptoms have resolved.

Cryptosporidium and Giardia in commercial and non-commercial oysters (Crassostrea gigas) and water from the Oosterschelde, The Netherlands.

The intestinal parasites Cryptosporidium and Giardia cause gastro-enteritis in humans and can be transmitted via contaminated water. Oysters are filter feeders that have been demonstrated to accumulate pathogens such as Salmonella, Vibrio, norovirus and Cryptosporidium from contaminated water and cause foodborne infections. Oysters are economically important shellfish that are generally consumed raw. Commercial and non-commercial oysters (Crassostrea gigas) and oyster culture water from the Oosterschelde, The Netherlands, were examined for the presence of Cryptosporidium oocysts and Giardia cysts. Nine of 133 (6.7%) oysters from two non-commercial harvesting sites contained Cryptosporidium, Giardia or both. Six of 46 (13.0%) commercial oysters harboured Cryptosporidium or Giardia in their intestines. Data on the viability of (oo)cysts recovered from Oosterschelde oysters were not obtained, however viable (oo)cysts were detected in surface waters that enter the Oosterschelde oyster harvesting areas. The detection of Cryptosporidium and Giardia in oysters destined for human consumption has implications for public health only when human pathogenic (oo)cysts that have preserved infectivity during their stay in a marine environment are present. Our data suggest that consumption of raw oysters from the Oosterschelde may occasionally lead to cases of gastro-intestinal illness.

Clinically relevant fungi in water and on surfaces in an indoor swimming pool facility.

The density of fungal contamination and the fungal diversity in an indoor swimming pool facility were assessed. A total of 16 surface samples and 6 water samples were analysed by using a combination of different (semi-) selective culture media. Isolated fungal colonies were identified to the genus or species level by sequencing of the internal transcribed spacer (ITS). The highest fungal counts in water and on surfaces were in the recreational pool (17CFU/100mL) and on a flexibeam (5.8CFU/cm2), respectively as compared with low counts (<0.1CFU/cm2) on the diving platform, bench tops and walls. The 357 obtained isolates belonged to 79 species and species complexes, 42 of which known as clinically relevant. Phialophora oxyspora (13.7%) and Phoma spp. (12.3%) were the most frequently identified groups. We demonstrated that despite chlorine treatment and regular cleaning of surfaces both water and surfaces were commonly infested with fungi, including many clinically relevant species.

Quantifying potential sources of surface water contamination with Campylobacter jejuni and Campylobacter coli.

Campylobacter is the most common causative agent of human bacterial gastroenteritis and is frequently found in surface water, where it indicates recent contamination with animal faeces, sewage effluent, and agricultural run-off. The contribution of different animal reservoirs to surface water contamination with Campylobacter is largely unknown. In the Netherlands, the massive poultry culling to control the 2003 avian influenza epidemic coincided with a 44-50% reduction in human campylobacteriosis cases in the culling areas, suggesting substantial environment-mediated spread of poultry-borne Campylobacter. We inferred the origin of surface water Campylobacter jejuni and Campylobacter coli strains in Luxembourg and the Netherlands, as defined by multilocus sequence typing, by comparison to strains from poultry, pigs, ruminants, and wild birds, using the asymmetric island model for source attribution. Most Luxembourgish water strains were attributed to wild birds (61.0%), followed by poultry (18.8%), ruminants (15.9%), and pigs (4.3%); whereas the Dutch water strains were mainly attributed to poultry (51.7%), wild birds (37.3%), ruminants (9.8%), and pigs (1.2%). Attributions varied over seasons and surface water types, and geographical variation in the relative contribution of poultry correlated with the magnitude of poultry production at either the national or provincial level, suggesting that environmental dissemination of Campylobacter from poultry farms and slaughterhouses can be substantial in poultry-rich regions.

Escherichia coli O157:H7 in drinking water from private water supplies in the Netherlands.

The microbiological quality of drinking water from 144 private water supplies in the Netherlands was tested and additionally the occurrence of Escherichia coli O157 was examined. Faecal indicators were enumerated by using standard membrane filtration methods. The presence of E. coli O157 was determined using a specific enrichment method. Eleven percent of the samples contained faecal indicators whereas E. coli O157:H7 was isolated from 2.7% of the samples that otherwise met the drinking water standards. The E. coli O157 positive water supplies were located on camp-sites in agricultural areas with large grazer densities. Pulsed field gel electrophoresis (PFGE) analysis suggested that cattle might have been the cause of contamination. Our results indicate that compliance with microbiological quality standards obtained in routine monitoring does not always guarantee the absence of pathogens. The presence of pathogens such as E. coli O157 may suggest possible health consequences; however, a risk assessment process should be performed as the monitoring of both faecal indicator parameters and pathogens do not predict the effect of microbial contamination of drinking water on a population.

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.

Distribution, Numbers, and Diversity of ESBL-Producing E. coli in the Poultry Farm Environment.

This study aimed to discern the contribution of poultry farms to the contamination of the environment with ESBL-producing Escherichia coli and therewith, potentially to the spread of these bacteria to humans and other animals. ESBL-producing E. coli were detected at all investigated laying hen farms (n = 5) and broiler farms (n = 3) in 65% (46/71) and 81% (57/70) of poultry faeces samples, respectively. They were detected in rinse water and run-off water (21/26; 81%), other farm animals (11/14; 79%), dust (21/35; 60%), surface water adjacent to farms (20/35; 57%), soil (48/87; 55%), on flies (11/73; 15%), and in barn air (2/33; 6%). The highest prevalence and concentrations in the outdoor environment were observed in soil of free-range areas at laying hen farms (100% of samples positive, geometric mean concentration 2.4×10(4) cfu/kg), and surface waters adjacent to broiler farms during, or shortly after, cleaning between production rounds (91% of samples positive, geometric mean concentration 1.9×10(2) cfu/l). The diversity of ESBL-producing E. coli variants with respect to sequence type, phylogenetic group, ESBL-genotype and antibiotic resistance profile was high, especially on broiler farms where on average 16 different variants were detected, and the average Simpson's Indices of diversity (SID; 1-D) were 0.93 and 0.94 among flock and environmental isolates respectively. At laying hen farms on average nine variants were detected, with SIDs of 0.63 (flock isolates) and 0.77 (environmental isolates). Sixty percent of environmental isolates were identical to flock isolates at the same farm. The highest proportions of 'flock variants' were observed in dust (94%), run-off gullies (82%), and barn air (67%), followed by surface water (57%), soil (56%), flies (50%) and other farm animals (35%).The introduction of ESBL-producing E. coli from poultry farms to the environment may pose a health risk if these bacteria reach places where people may become exposed.

[Infections following recreational activities in lakes, rivers and canals: present and future risks of transmission in the Netherlands]. / Infecties door recreatie in oppervlaktewater: huidige en toekomstige risico's op transmissie in Nederland.

Poor microbiological quality of surface water such as lakes, rivers and canals used for recreational purposes by large groups of people may lead to many cases of disease. Microbiological contamination can originate from various sources, including both human and animal faeces, but can also result from increased levels of naturally occurring microorganisms. In Europe, the European Bathing Water Directive applies to all official bathing sites. At these locations, water quality must be checked regularly and must meet requirements for maximum faecal contamination. However, the most common recreational water-related health conditions in the Netherlands are skin complaints, predominantly 'swimmers itch'; this is caused by Trichobilharzia, a parasite of non-faecal origin. The number of disease outbreaks in any summer is strongly linked to the number of days when the maximum temperature is 25°C (77°F ) or above. Besides the impact of climate on recreational water-related pathogens, human behaviour also plays a role; the outcome of the interaction between these and other factors is as yet uncertain.