Changes of the bacterial assemblages throughout an urban drinking water distribution system

We analyzed the bacterial 16S rRNA gene diversity throughout the major components of the drinking water distribution system of a ca. 52,000-inhabitants city (Trikala City, Greece) in order to describe the changes of the bacterial assemblages and to detect possible bacterial pathogens which are not included in the standard monitoring process. Bacterial DAPI counts and DNA extraction was performed in the water pumping wells, the water treatment tank and tap water from households. Approximately 920 bp of the bacterial 16S rDNA were PCR-amplified, cloned, and sequenced for a total of 191 clones, which belonged to 112 unique phylotypes. The water of the pumping wells harbored a typical subsurface bacterial assemblage, with no human pathogens, dominated by β-Proteobacteria. Cell abundance in the water treatment tank decreased significantly, close to detection limit, but bacterial diversity remained high. However, the dominance of β-Proteobacteria decreased considerably, indicating the sensitivity of this group to drinking water disinfection treatment. Tap water from the households hosted a much less diverse, low-cell bacterial assemblage, dominated by Mycobacterium-like phylotypes, related to biofilm bacterial communities.     

Numerical Modeling on the Effect of Dissolved Oxygen on Nitrogen Transformation and Transport in Unsaturated Porous System

Nitrogen pollution in groundwater resulting from wastewater application to land is a common problem, and it causes a major threat to groundwater-based drinking water supplies. In this study, a numerical model is developed to study the nitrogen species transport and transformation in unsaturated porous media. Further, a new mass transfer module for dissolved oxygen (DO) is incorporated in the one-dimensional numerical model for nitrogen species transport to describe the fate and transport of nitrogen species, dissolved oxygen, dissolved organic carbon (DOC), and biomass. The spatial and temporal variation of dissolved oxygen is incorporated in the model through the mass transfer from gaseous phase to water phase in an unsaturated porous system. The numerical results of the water flow model and single species and multispecies transport model in an unsaturated zone developed for this purpose have been validated with the available analytical/numerical solution. The developed model is applied in clay loam, silt, and sand soils to analyze the transport behavior of nitrogen species under unsaturated condition. The numerical results suggest that the high rate of oxygen mass transfer from the air phase to the water phase positively increases the dissolved oxygen in the applied wastewater and enhances the nitrification process. Because of this high oxygen mass transfer, the nitrate nitrogen concentration significantly increases in the unsaturated zone and the same is transported to a larger depth at higher simulation period. On the other hand, the low rate of oxygen mass transfer implicitly enhances the denitrification process and finally reduces the nitrate nitrogen concentration in the unsaturated zone. The numerical results also show that the nitrate nitrogen transport is rapid in sandy soil when compared with clay loam and silty soils under high oxygen mass transfer rate. In essence, the high oxygen mass transfer rate significantly increases the nitrate nitrogen in the unsaturated zone, especially at a greater depth at larger time levels and eventually affects the groundwater quality.     

Antibiotic resistance profile of bacterial isolates from animal farming aquatic environments and meats in a peri-urban community in Daejeon, Korea

The increasing usage of antibiotics in the animal farming industry is an emerging worldwide problem contributing to the development of antibiotic resistance. The purpose of this work was to investigate the prevalence and antibiotic resistance profile of bacterial isolates collected from animal farming aquatic environments and meats in a peri-urban community in Daejeon, Korea. In an antibacterial susceptibility test, the bacterial isolates showed a high incidence of resistance (～26.04%) to cefazolin, tetracycline, gentamycin, norfloxacin, erythromycin and vancomycin. The results from a test for multiple antibiotic resistance indicated that the isolates were displaying an approximately 5-fold increase in the incidence of multiple antibiotic resistance to combinations of two different antibiotics compared to combinations of three or more antibiotics. Most of the isolates showed multi-antibiotic resistance, and the resistance patterns were similar among the sampling groups. Sequencing data analysis of 16S rRNA showed that most of the resistant isolates appeared to be dominated by the classes Betaproteobacteria and Gammaproteobacteria, including the genera Delftia, Burkholderia, Escherichia, Enterobacter, Acinetobacter, Shigella and Pseudomonas.     

Determination of an acceptable assimilable organic carbon (AOC) level for biological stability in water distribution systems with minimized chlorine residual

There is considerable interest in minimizing the chlorine residual in Japan because of increasing complaints about a chlorinous odor in drinking water. However, minimizing the chlorine residual causes the microbiological water quality to deteriorate, and stricter control of biodegradable organics in finished water is thus needed to maintain biological stability during water distribution. In this investigation, an acceptable level of assimilable organic carbon (AOC) for biologically stable water with minimized chlorine residual was determined based on the relationship between AOC, the chlorine residual, and bacterial regrowth. In order to prepare water samples containing lower AOC, the fractions of AOC and biodegradable organic matter (BOM) in tap water samples were reduced by converting into biomass after thermal hydrolysis of BOM at alkaline conditions. The batch-mode incubations at different conditions of AOC and chlorine residual were carried out at 20°C, and the presence or absence of bacterial regrowth was determined. The determined curve for biologically stable water indicated that the acceptable AOC was 10.9 μg C/L at a minimized chlorine residual (0.05 mg Cl₂/L). This result indicated that AOC removal during current water treatment processes in Japan should be significantly enhanced prior to minimization of the chlorine residual in water distribution.     

Bacterial communities in urban aerosols collected with wetted-wall cyclonic samplers and seasonal fluctuations of live and culturable airborne bacteria

Airborne transmission of bacterial pathogens from point sources (e.g., ranches, dairy waste treatment facilities) to areas of food production (farms) has been suspected. Determining the incidence, transport and viability of extremely low levels of pathogens require collection of high volumes of air and characterization of live bacteria from aerosols. We monitored the numbers of culturable bacteria in urban aerosols on 21 separate days during a 9 month period using high volume cyclonic samplers at an elevation of 6 m above ground level. Culturable bacteria in aerosols fluctuated from 3 CFU to 6 million CFU/L of air per hour and correlated significantly with changes in seasonal temperatures, but not with humidity or wind speed. Concentrations of viable bacteria determined by fluorescence staining and flow cytometry correlated significantly with culturable bacteria. Members of the phylum Proteobacteria constituted 98% of the bacterial community, which was characterized using 16S rRNA gene sequencing using DNA from aerosols. Aquabacterium sp., previously characterized from aquatic environments, represented 63% of all clones and the second most common were Burkholderia sp; these are ubiquitous in nature and some are potential human pathogens. Whole genome amplification prior to sequencing resulted in a substantial decrease in species diversity compared to characterizing culturable bacteria sorted by flow cytometry based on scatter signals. Although 27 isolated colonies were characterized, we were able to culture 38% of bacteria characterized by sequencing. The whole genome amplification method amplified DNA preferentially from Phyllobacterium myrsinacearum, a minor member of the bacterial communities, whereas Variovorax paradoxus dominated the cultured organisms.     

Occurrence of antibiotic resistance and characterization of resistance genes and integrons in Enterobacteriaceae isolated from integrated fish farms in South China

Antibiotics are still widely applied in animal husbandry to prevent diseases and used as feed additives to promote animal growth. This could result in antibiotic resistance to bacteria and antibiotic residues in animals. In this paper, Enterobacteriaceae isolated from four integrated fish farms in Zhongshan, South China were tested for antibiotic resistance, tetracycline resistance genes, sulfonamide resistance genes, and class 1 integrons. The Kirby-Bauer disk diffusion method and polymerase chain reaction (PCR) assays were carried out to test antibiotic susceptibility and resistance genes, respectively. Relatively high antibiotic resistance frequencies were found, especially for ampicillin (80%), tetracycline (52%), and trimethoprim (50%). Out of 203 Enterobacteriaceae isolates, 98.5% were resistant to one or more antibiotics tested. Multiple antibiotic resistance (MAR) was found highest in animal manures with a MAR index of 0.56. Tetracycline resistance genes (tet(A), tet(C)) and sulfonamide resistance genes (sul2) were detected in more than 50% of the isolates. The intI1 gene was found in 170 isolates (83.7%). Both classic and non-classic class 1 integrons were found. Four genes, aadA5, aadA22, dfr2, and dfrA17, were detected. To our knowledge, this is the first report for molecular characterization of antibiotic resistance genes in Enterobacteriaceae isolated from integrated fish farms in China and the first time that gene cassette array dfrA17-aadA5 has been detected in such fish farms. Results of this study indicated that fish farms may be a reservoir of highly diverse and abundant antibiotic resistant genes and gene cassettes. Integrons may play a key role in multiple antibiotic resistances posing potential health risks to the general public and aquaculture.     

Evaluating seasonal dynamics of bacterial communities in marine fish aquaculture: a preliminary study before applying phage therapy

The increasing problem of antibiotic resistance in common pathogenic bacteria and the concern about the spreading of antibiotics in the environment bring the need to find new methods to control fish pathogens. Phage therapy represents a potential alternative to antibiotics, but its use in aquaculture requires a detailed understanding of bacterial communities, namely of fish pathogenic bacteria. Therefore, in this study the seasonal dynamics of the overall bacterial communities, microbiological water quality and disease-causing bacteria were followed in a marine aquaculture system of Ria de Aveiro (Portugal). Analysis of the bacterial diversity of the water samples by denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments indicates that the bacterial community structure varied seasonally, showing a higher complexity during the warm season. The diversity of the main fish pathogenic bacteria, assessed by DGGE targeting the Vibrio genus, showed lower seasonal variation, with new dominating populations appearing mainly in the spring. Bacterial indicators, faecal coliforms and enterococci, enumerated by the filter-membrane method, also varied seasonally. The fluorescent in situ hybridization (FISH) results showed that the specific groups of bacteria varied during the study period and that the non-indigenous Enterobactereaceae family was the most abundant group followed by Vibrio and Aeromonas. The seasonal variation detected in terms of density and structure of total and pathogenic bacterial communities demonstrates the need for a careful monitoring of water through the year in order to select the suitable phages to inactivate fish pathogenic bacteria. The spring season seems to be the critical time period when phage therapy should be applied.     

Contact Tank Design Impact on Process Performance

In this study three-dimensional numerical models were refined to predict reactive processes in disinfection contact tanks (CTs). The methodology departs from the traditional performance assessment of contact tanks via hydraulic efficiency indicators, as it simulates directly transport and decay of the disinfectant, inactivation of pathogens and accumulation of by-products. The method is applied to study the effects of inlet and compartment design on contact tank performance, with special emphasis on turbulent mixing and minimisation of internal recirculation and short-circuiting. In contrast to the conventional approach of maximising the length-to-width ratio, the proposed design changes are aimed at addressing and mitigating adverse hydrodynamic structures, which have historically led to poor hydraulic efficiency in many existing contact tanks. The results suggest that water treatment facilities can benefit from in-depth analyses of the flow and kinetic processes through computational fluid dynamics, resulting in up to 38 % more efficient pathogen inactivation and 14 % less disinfection by-product formation.     

Characterisation of Indicator Organisms and Pathogens in Domestic Greywater for Recycling

Greywater from baths, showers and washbasins was collected separately from all other domestic wastewater at a university block of flats with a dual reticulation system and analysed for a range of contaminants including indicator organisms and pathogens. Greywater flow and temperature were also monitored and a diurnal variation was observed. Physical and chemical water quality parameters were similar to previously published data, although measured COD and BOD levels appeared to be lower, possibly due to settlement or biodegradation in the storage tanks. Plate counts and indicator organism concentrations were consistently high suggesting a high level of human bacterial contamination necessitating biological treatment and disinfection if the water is to be used for recycling. However, these high levels of indicator organisms did not correlate to pathogen presence and should not be used as pathogen indicators in greywater. One positive count of Salmonella veltereden was observed as well as low levels of Giardia. Cryptosporidium, Escherichia coli O157:H7, enteroviruses and Legionella were not identified in any of the samples. The research also highlighted a number of problems with the complexity of this type of sampling programme, such as identifying the most likely time to isolate pathogens and analysing an ‘unusual’ water source.     

Limnochemistry and nutrient dynamics in Upper Lake, Bhopal, India

In limnetic ecosystem, water quality depends upon physical, chemical, and biological factors. Effects of temperature, light scattering, and absorption by suspended and dissolved matter, transport, and mixing of nutrients within the lake are the significant factors as far as water quality is concerned. Nutrient loading into the lake and internal cycling of nutrients is always a matter of concern and critical to number of processes. During the winter season, heat and momentum transfer at the lake surface and the temperature-density relation of water destabilize the water column and drive vertical mixing and transport processes. The deepening of the surface layer produces nutrient transfer from the hypolimnion into the euphotic zone of epilimnion. It may also resuspend sediments that would have settled under stratified conditions, or redistribute particles that may still be in suspension. Thus, there exists a complex connection between the hydrodynamics and water quality issues. Present study is an effort to understand how seasonal changes in the limnetic ecosystem regulate the limnochemistry and movement of nutrient. The study revealed that significant variations of nutrients and organic load were observed between epilimnion and hypolimnion during summer season, and the lake was found in hyper-eutrophic condition throughout the study period.     

Total organic carbon analyzers as tools for measuring carbonaceous matter in natural waters

For some utilities, new US drinking water regulations may require the removal of disinfection byproduct (DBP) precursor material as a means of minimizing DBP formation. The Environmental Protection Agency's Stage 1 DBP Rule relies on total organic carbon (TOC) concentrations as a measure of the effectiveness of treatment techniques for removing organic material that could act as DBP precursors. Accordingly, precise and accurate methods are needed for the determination of TOC and dissolved organic carbon (DOC) concentrations in raw and finished potable water supplies. This review describes the current analytical technologies and summarizes the key factors affecting measurement quality. It provides a look into the fundamental principles and workings of TOC analyzers. Current peroxydisulfuric acid wet ashing methods and combustion methods are discussed. Issues affecting quality control, such as non-zero blanks and preservation, are covered. Some of the difficulties in analyzing water for TOC and DOC that were identified up to 20 years ago still remain problematic today. Limitations in technology, reagent purity, operator skill and knowledge of natural organic matter (NOM) can preclude the level of precision and accuracy desirable for compliance monitoring.     

Destruction of fecal bacteria in wastewater by three photosensitizers

Agriculture in Tunisia faces acute problems of water quality and quantity caused by limited conventional water resources. One possibility to cope with low water resources is to purify wastewater for reuse. Among different methods for the disinfection of wastewater, the inactivation of fecal coliforms using a combination of a photosensitiser (Rose Bengal, Methylene Blue, cationic porphyrin) with sunlight was determined on a small scale. In parallel the sensitizer photobleaching was also followed under the same conditions. The results described in this paper show that the meso-substituted cationic porphyrin is more efficient and more photostable than Methylene Blue and Rose Bengal in wastewater. A lower cationic porphyrin concentration, 1 microM, resulted in very little cell death. Higher concentrations, 5 microM or 10 microM, produced more cell death. Nevertheless there was a small difference between concentrations 5 and 10 microM. By increasing the duration of irradiation we can improve the log reduction in bacteria and compensate for a low concentration of sensitizer or for a less efficient type of sensitizer. The same log reduction in fecal bacteria was obtained with 5 and 10 microM of cationic porphyrin during the fourth hour of treatment.     

Whole-cell bacterial biosensors for rapid and effective monitoring of heavy metals and inorganic pollutants in wastewater

The increasing number of potentially harmful pollutants in the wastewater effluent discharge necessitates the need for the development of fast and cost effective analytical techniques for extensive monitoring programmes to assess the effectiveness of the treatment process. This study compared the use of bacterial biosensors to the conventional Daphnia magna assay, Chemical Oxygen Demand (COD) and Biochemical Oxygen Demand (BOD) tests as well as chemical analysis, for monitoring the toxicity of wastewater. The bacterial biosensors constructed in this study, using S. sonnei and E. coli, were found to be sensitive to the toxicity of the wastewater effluents. A linear increase in bioluminescence with increasing concentration of heavy metals and inorganic pollutants in water was observed, with a correlation coefficient (r(2)) as high as 0.995 and 0.997, respectively. No notable correlation between biosensor toxicity and BOD and COD test results was observed. These bacterial biosensors could provide appropriate alternatives for a rapid, sensitive and cost effective detection of wastewater quality. However, the differences in sensitivity obtained for the different systems suggest that the use of a battery of toxicity assays may be required to provide a real ecotoxicological assessment of wastewater samples.     

Assessing the feasibility of wastewater recycling and treatment efficiency of wastewater treatment units

Wastewater reuse can significantly reduce environmental pollution and save the water sources. The study selected Cheng-Ching Lake water treatment plant in southern Taiwan to discuss the feasibility of wastewater recycling and treatment efficiency of wastewater treatment units. The treatment units of this plant include wastewater basin, sedimentation basin, sludge thickener and sludge dewatering facility. In this study, the treatment efficiency of SS and turbidity were 48.35–99.68% and 24.15–99.36%, respectively, showing the significant removal efficiency of the wastewater process. However, the removal efficiencies of NH₃–N, total organic carbon (TOC) and chemical oxygen demand (COD) are limited by wastewater treatment processes. Because NH₃–N, TOC and COD of the mixing supernatant and raw water are regulated raw water quality standards, supernatant reuse is feasible and workable during wastewater processes at this plant. Overall, analytical results indicated that supernatant reuse is feasible.     

Removal and recovery of metals from a coal pile runoff

The removal and recovery of heavy metals from a coal pilerunoff water using a mixture of multiple metal-tolerantbacterial strains of ATCC 55673, and ATCC 55674 and a Pseudomonas sp. was investigated. The analysis of elementalcomposition of metal precipitates recovered from the bacterialbiomass by transmission electron microscopy and energy dispersiveX-ray analysis revealed the presence of metals originally presentin the wastewater. In addition, analysis of metals in culturesupernatant and bacterial biomass by inductively coupled plasmaemission spectroscopy (ICP-ES) indicated a removal range of 82-100% and a recovery of 15-58% of metals from the wastewater and bacterial biomass, respectively.     

印染废水处理污泥危险废物鉴别过程中有毒污染物的迁移转化及鉴别因子筛选

简述了印染废水处理污泥危险废物鉴别过程中有毒污染物的迁移转化及鉴别因子筛选。以采用活性染料的印染企业为例,从常用的原辅材料、使用的生产工艺、常用的废水处理工艺等几个方面来对印染废水处理污泥危险废物鉴别过程中有毒污染物的迁移及转化进行分析。提出危险废物鉴别工作任重而道远,危险废物鉴别的关键在于日常监管。     

Assessment of physicochemical parameters and prevalence of virulent and multiple-antibiotic-resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> in treated effluent of two wastewater treatment plants and receiving aquatic <Emphasis Type="Italic">milieu</Emphasis> in Durban,South Africa

The poor operational status of some wastewater treatment plants often result in the discharge of inadequately treated effluent into receiving surface waters. This is of significant public health concern as there are many informal settlement dwellers (ISDs) that rely on these surface waters for their domestic use. This study investigated the treatment efficiency of two independent wastewater treatment plants (WWTPs) in Durban, South Africa and determined the impact of treated effluent discharge on the physicochemical and microbial quality of the receiving water bodies over a 6-month period. Presumptive Escherichia coli isolates were identified using biochemical tests and detection of the mdh gene via PCR. Six major virulence genes namely eae, hly, fliC, stx1, stx2, and rfbE were also detected via PCR while antibiotic resistance profiles of the isolates were determined using Kirby-Bauer disc diffusion assay. The physicochemical parameters of the wastewater samples ranged variously between 9 and 313.33 mg/L, 1.52 and 76.43 NTUs, and 6.30 and 7.87 for COD, turbidity, and pH respectively, while the E. coli counts ranged between 0 and 31.2?×?10³ CFU/ml. Of the 200 selected E. coli isolates, the hly gene was found in 28 %, fliC in 20 %, stx2 in 17 %, eae in 14 %, with stx1 and rfbE in only 4 % of the isolates. Notable resistance was observed toward trimethoprim (97 %), tetracycline (56 %), and ampicillin (52.5 %). These results further highlight the poor operational status of these WWTPs and outline the need for improved water quality monitoring and enforcement of stringent guidelines.     

Incidence of metal and antibiotic resistance in Pseudomonas spp. from the river water, agricultural soil irrigated with wastewater and groundwater

A total of 144 isolates of Pseudomonas spp. (48 each from the Yamuna River water, wastewater irrigated soil and groundwater irrigated soil) were tested for their resistance against certain heavy metals and antibiotics. Minimum inhibitory concentrations (MICs) of Hg^2?+?, Cd^2?+?, Cu^2?+?, Zn^2?+?, Ni^2?+?, Pb^2?+?, Cr^3?+? and Cr^6?+? for each isolate were also determined. A maximum MIC of 200 ??g/ml for mercury and 3,200 ??g/ml for other metals were observed. The incidences of metal resistance and MICs of metals for Pseudomonas isolates from the Yamuna water and wastewater irrigated soil were significantly different to those of groundwater irrigated soil. A high level of resistance against tetracycline and polymyxin B (81.2%) was observed in river water isolates. However, 87.5% of Pseudomonas isolates from soil irrigated with wastewater showed resistance to sulphadiazine, whereas 79.1% were resistant to both ampicillin and erythromycin. Isolates from soil irrigated with groundwater exhibited less resistance towards heavy metals and antibiotics as compared to those of river water and wastewater irrigated soil. Majority of the Pseudomonas isolates from water and soil exhibited resistance to multiple metals and antibiotics. Resistance was transferable to recipient Escherichia coli AB2200 strains by conjugation. Plasmids were cured with the curing agent ethidium bromide and acridine orange at sub-MIC concentration.     

Comparison of the microbial load of incoming and distal outlet waters from dental unit water systems in Istanbul

This is a cross-sectional study of the incoming and distal outlet water quality from 41 dental units in Istanbul, carried out to compare the total microbial loads using traditional culture method versus epifluorescence microscopy. The possible presence of Legionella pneumophila using traditional culture method was also analyzed. One hundred and twenty three samples were taken from the high-speed handpiece lines, air-water syringe lines and source (incoming) water supplies from 41 dental units. The samples were assayed for live/dead bacteria, heterotrophic bacterial counts and presence of L. pneumophila bacteria. Thirty nine out of 41 dental units (91%) were not able to meet the standard limit of 200 CFU/ml in dental unit waters. The live bacterial counts were 1-1.5 orders of magnitude higher than aerobic mesophilic heterotrophic bacteria. L. pneumophila (serogroup 2-14) was isolated from five out of 41 units. Some dental units were using commercially bottled (19 l) drinking water as a source. The source water of eight dental unit was heavily contaminated which were fed up by commercially bottled drinking water.     

Waterborne outbreaks in diarrhoea endemic foci of India: a longitudinal exploration and its implications

Diarrhoea remains a global public health enigma raising deep concerns for the health planners since contaminated potable water often spoils the community health structure. We hereby report a 6-year odd continuing outbreak surveillance report based on potable water indices, during which 264 water samples were screened from different districts of West Bengal, India. Samples were analysed for the presence of different enteropathogenic bacterial species by conventional molecular tools and their sensitivity to antibiotics. 78.03% samples were positive for enteropathogenic bacterial organisms and 75% samples harbored Coliform. 45.45, 12.12, and 4.16% samples were positive for E.coli, V. cholerae, and V. mimicus, respectively. Diarrhoeagenic E.coli 7 EPEC, 10 ETEC, and 2 EIEC were isolated along with 2 V. cholerae O1 Ogawa (ctxA and tcpA ElTor positive), one each from tube well and pond. Interestingly, 4 V.cholerae non-O1/non-O139 also harbored hlyA gene. The detection of toxin genes among this bacterial pool of sampled water indicates the fallout of the potable water sources, thus enabling us to establish that it is none other than the contaminated potable water system which often wreaks havoc in the south Bengal diarrhoeal menace. The consequences are further complicated by the presence of drug-resistant pathogenic bacterial pool to fluoroquinolone, beta-lactams, and cephalosporins, in the accessible potable water, with threats of outbreaks exploding into an epidemic, given suitable environment, poor sanitation, and unhygienic practices. Therefore, we strongly recommend re-modelling of ‘point-of-use water disinfection’ measures and adequate personal hygiene for healthier community life.