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Antibiotic resistance among gram-negative bacteria is increasingly becoming a problem of global concern. Particularly problematic is the emergence of resistance to last-resort antibiotics such as carbapenems and colistin. The increasing number of reports on the plasmid-mediated colistin resistance gene mcr-1 in isolates worldwide is raising concerns for the future usefulness of this class of antibiotics. Dissemination of mcr-1 is believed to have originated mainly from animal breeding, however, the role of the environment as a transmission source is not yet fully understood. In the current study, 89 extended-spectrum β-lactamase-producing Escherichia coli isolated from 231 samples from different environmental sources in 12 villages in a rural area of Shandong, China, were screened for mcr-1. 17 (19.1%) mcr-1 -positive isolates were found from different environmental sources, aggregated in 6 villages. Plasmids of three different Inc-groups carrying mcr-1 were confirmed, indicating that the widespread geographical distribution of mcr-1 in the local area is due to a number of different plasmids. Additionally, almost a third (29.4%) of the isolates carried virulence factors associated to intestinal pathogenic E. coli. These results illustrate the high complexity of the transmission patterns of mcr-1 among different environmental matrices on a local scale and the potential for the environment to facilitate dissemination and emergence of antibiotic-resistant and virulent strains of bacteria. Image 1 • In rural China, the mcr-1 contamination was serious. • mcr-1 can be transferred cross environmental matrices through the plasmid. • Pathogenic E. coli with mcr-1 were found and indicate a potential health risk. [ABSTRACT FROM AUTHOR]

The aims of the study were to determine the survival of Escherichia coli O157 on lettuce as a function of temperature and light intensity, and to use that information in a screening-level quantitative microbial risk assessment (QMRA) in order to evaluate risk-reducing strategies including irrigation water quality guidelines, rinsing, and holding time between last irrigation and harvest. Iceberg lettuce was grown in a climate chamber and inoculated with E. coli O157. Bacterial numbers were determined with the standard plate count method after inoculation and 1, 2, 4, and 7 day(s) postinoculation. The experiments were carried out at 11, 18, and 25°C in light intensities of 0, 400, and 600 mmol (m²)-1s-1, There was a significant effect of temperature and light intensity on survival, with less bacteria isolated from lettuce incubated at 25 and 18°C compared with 11°C (P < 0.0001), and in light intensities of 400 and 600 mmol (m²)-1s-1 compared with 0 mmol (m²)-1s-1 (P < 0.001). The average log reductions after 1, 2, 4, and 7 day(s) were 1.14, 1.71, 2.04, and 3.0, respectively. The QMRA compared the relative risk with lettuce consumption from 20 scenarios. A stricter water quality guideline gave a mean fivefold risk reduction. Holding times of 1, 2, 4, and 7 day(s) reduced the risk 3, 8, 8, and 18 times, respectively, compared with harvest the same day as the last irrigation. Finally, rinsing lettuce for 15 s in cold tap water prior to consumption gave a sixfold risk reduction compared with eating unrinsed lettuce. Sensitivity analyses indicated that variation in bacterial inactivation had the most significant effect on the risk outcome. A QMRA determining the relative risks between scenarios reduces uncertainty and can provide risk managers with decision support. [ABSTRACT FROM AUTHOR]

The faecal contamination of greywater in a local treatment system at Vibya˚sen, north of Stockholm, Sweden was quantified using faecal indicator bacteria and chemical biomarkers. Bacterial indicator densities overestimated the faecal load by 100–1000-fold when compared to chemical biomarkers. Based on measured levels of coprostanol, the faecal load was estimated to be 0.04 g person−1 day−1. Prevalence of pathogens in the population and the faecal load were used to form the basis of a screening-level quantitative microbial risk assessment (QMRA) that was undertaken for rotavirus, Salmonella typhimurium, Campylobacter jejuni, Giardia lamblia and Cryptosporidium parvum. The different exposure scenarios simulated—direct contact, irrigation of sport fields and groundwater recharge—gave unacceptably high rotavirus risks (0.04) despite a low faecal load. The poor reduction of somatic coliphages, which were used as a virus model, in the treatment was one main reason and additional treatment of the greywater is suggested. Somatic coliphages can under extreme circumstances replicate in the wastewater treatment system and thereby underestimate the virus reduction. An alternative QMRA method based on faecal enterococci densities estimated similar risks as for rotavirus. Growth conditions for Salmonella in greywater sediments were also investigated and risk modelling based on replication in the system increased the probability of infection from Salmonella 1000-fold, but it was still lower than the risk of a rotavirus infection. [Copyright &y& Elsevier]

Monitoring of faecal indicator organisms, such as Escherichia coli , in environmental and drinking waters is inadequate for the protection public health, primarily due to the poor relationship between E. coli and the occurrence of human pathogens, especially viruses, in environmental samples. Nevertheless, measurements of faecal indicator organisms within the risk based approach, can provide valuable information related to the magnitude and variability of faecal contamination, and hence provide insight into the expected level of potential pathogen contamination. In this study, a modelling approach is presented that estimates the concentration of norovirus in surface water relying on indicator monitoring data, combined with specific assumptions regarding the source of faecal contamination. The model is applied to a case study on drinking water treatment intake from the Glomma River in Norway. Norovirus concentrations were estimated in two sewage sources discharging into the river upstream of the drinking water offtake, and at the source water intake itself. The characteristics of the assumed source of faecal contamination, including the norovirus prevalence in the community, the size of the contributing population and the relative treatment efficacy for indicators and pathogens in the sewage treatment plant, influenced the magnitude and variability in the estimated norovirus concentration in surface waters. The modelling exercise presented is not intended to replace pathogen enumeration from environmental samples, but rather is proposed as a complement to better understand the sources and drivers of viruses in surface waters. The approach has the potential to inform sampling regimes by identifying when the best time would be to collect environmental samples; fill in the gaps between sparse datasets; and potentially extrapolate existing datasets in order to model rarer events such as an outbreak in the contributing population. In addition, and perhaps most universally, in the absence of pathogen data, this approach can be used as a first step to predict the source water pathogen concentration under different contamination scenarios for the purpose of quantifying microbial risks. [ABSTRACT FROM AUTHOR]

Objectives To investigate the occurrence of antibiotic residues in different types of environmental samples including water samples in rural Shandong province, China. Further, to characterize the potential ecological risk for development of antibiotic resistance in the environment, and the potential direct human health risk of exposure to antibiotics via drinking water and vegetables. Methods Environmental samples ( n  = 214) (river water, waste water, drinking water, sediments, manure, soil and edible parts of vegetables) were collected in twelve villages in Shandong province in eastern China. High performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) was used to determine the concentration of antibiotic residues. The ratio of the measured environmental concentrations (MEC) to the predicted no-effect concentrations (PNEC) was used to evaluate the ecological risk (risk quotient, RQ) for development of antibiotic resistance. The potential risks to human health through exposure to antibiotics in drinking water were assessed by comparing measured environmental concentrations (MEC) and predicted no-effect concentration in drinking water (PNEC DW ), and in vegetables by comparing estimated daily intake (EDI) to ADI. Results Sulfapyridine, sulfamethoxazole, ciprofloxacin, enrofloxacin, levofloxacin, norfloxacin, chloramphenicol, florfenicol, doxycycline, and metronidazole were detected at concentrations ranging between 0.3 and 3.9 ng/L in river water, 1.3 and 12.5 ng/L in waste water, 0.5 and 21.4 ng/L in drinking water, 0.31 and 1.21 μg/kg in river sediment, 0.82 and 1.91 μg/kg in pig manure, 0.1 and 11.68 μg/kg in outlet sediment, 0.5 and 2.5 μg/kg in soil, and 6.3 and 27.2 μg/kg in vegetables. The RQs for resistance development were >1 for enrofloxacin, levofloxacin, and ranged between 0.1 and 1 for ciprofloxacin. MECs/PNEC DW ratios were <1 from exposure to antibiotics through drinking water for both adults and children. EDI/ADI ratios were <0.1 from exposure to antibiotics by vegetable consumption. Conclusions Antibiotic pollutants were ubiquitous in various environmental compartments of Shandong province of China. Risk estimates indicated a potential for the measured levels of enrofloxacin, levofloxacin and ciprofloxacin in waste water to pose an ecological risk for resistance selection, and further studies are needed to validate this finding. The investigated antibiotics did not appear to pose an appreciable direct human health risk from environmental exposure through drinking water or vegetables consumption. However, they might still pose a risk for resistance development. [ABSTRACT FROM AUTHOR]

On-farm measures can be used in multi-barrier schemes to manage microbial risks from consumption of wastewater-irrigated vegetables, especially where informality of the practice determines minimal external support for farmers. Evidence indicates that cessation of irrigation greatly reduces microbial contamination on leafy vegetables, but at the expense of produce quality. Replacing wastewater with higher-quality irrigation water during the last days of cultivation is an alternative to cessation of irrigation that does not compromise produce quality. This study evaluated the effect of wastewater substitution under on-farm conditions on different indicators of microbial contamination of lettuce. Lettuce was cultivated in experimental plots and irrigated with three water sources: spring water, water from a wastewater-polluted river and effluent from a primary wastewater treatment plant, but with the river water replaced by spring water in half the plots about two weeks before harvest. The experiment was repeated four times in different seasons. Irrigation water samples collected during cultivation and lettuce samples collected at harvest were analysed for helminth eggs, Escherichia coli and coliphages. Variables characterizing the irrigation practices and environmental conditions were recorded. There were no significant differences in helminth egg or E. coli concentrations on lettuce (medians ranged from −0.7 to −0.1 log 10 eggs g−1 and 0.6–1.4 log 10 cfu g−1, respectively) between any of the treatments involving wastewater irrigation; no statistical analysis was possible for coliphages because concentrations on lettuce were mostly at or below the detection limit (94% of samples). Variables associated with temperature and soil explained helminth egg and E. coli concentrations on lettuce, while number of days of irrigation with spring water (representing wastewater substitution) was significant only for E. coli. It was concluded that the experimental conditions were suboptimal for successful implementation of wastewater substitution for on-farm microbial risk management, but key variables for successful implementation were identified. • Extended die-off was studied for microbial reduction on wastewater-irrigated crop. • 11–14 days of die-off were achieved by replacing wastewater with a cleaner source. • Water, soil, and lettuce samples were taken during four crop cycles. • Studied microbes were reduced in < 1 log 10 by irrigation water substitution. • Reduction rates may be higher in warmer regions/with higher irrigation efficiencies. [ABSTRACT FROM AUTHOR]