Characteristics of indicator microorganisms in paddy rice plots after reclaimed water irrigation.

A study was performed to examine the effects of reclaimed-water irrigation on microorganism concentration in the ponded-water of paddy rice plots. Several treatments were used and each one was triplicated to evaluate the change of indicator microorganisms (total coliforms (TC), faecal coliforms (FC), and E. coli) concentrations in 2003 and 2004 growing seasons. Their concentrations increased significantly after irrigation, but decreased by approximately 45% in 24 h. It implies that agricultural activities such as ploughing and fertilising need to be practiced one or two days after irrigation, considering health risks. Treatments with UV-disinfected water irrigation, demonstrated significantly lower concentrations than others, including control plots where natural water was irrigated. The monitoring result from actual paddy rice fields and experimental paddy plots showed that concentrations of indicator microorganisms ranged from 10(2) to 10(5) MPN/100 mL. The field scale water reuse research project is in progress and national guidelines for reclaimed-water irrigation are under preparation in Korea. A comprehensive assessment of existing agricultural practices and a thorough monitoring in the fields as well as treatment plots are recommended to make national guidelines more representative.     

Stormflow-dominated loads of faecal pollution from an intensively dairy-farmed catchment

Rainstorms can flush large amounts of faecal pollution from land sources into water bodies, threatening, particularly, contact recreation and bivalve shellfish harvest. We quantified the faecal pollution loads of stormflows in the Toenepi Stream, draining a catchment in intensive dairy-farming (Waikato Region, New Zealand). In this stream, as is typical, E. coli concentration peaks well ahead of flow on storm flow hydrographs, which complicates calculation of loads. However, stormflow E. coli concentration correlates with turbidity in the Toenepi Stream, so we used a continuously-recording turbidimeter to estimate 'continuous' E. coli concentrations and thence E. coli fluxes (cfu/s) and loads (cfu). E. coli was measured on 25 out of the 30 (83%) of storm events occurring in the Toenepi Stream in a 12-month period, using an automatic sampler sampling every 2 hrs over stormflow hydrographs for microbial analysis (within 48 hr). E. coli (cfu) yield on individual events tended to increase systematically with event size. The sum of storm-flow exports (occurring 24% of total time) amounted to 95% of the total annual E. coli export from the Toenepi Catchment. The stream exported about 6% of the (expected) total E. coli production in cattle faeces within the catchment.     

The utility of the AC/TC ratio for watershed management: a case study.

A human-impacted watershed was monitored during the dry summer seasons in 2002 and 2003 to investigate the impact of providing access to sewer mains to local village residences. Faecal coliform concentrations were monitored at select sites along the 30-mile stretch of creek, together with faecal streptococci, enterococci and total coliforms. Analysis of the results found that levels of faecal coliforms were inadequate at identifying significant known influxes of human and animal sewage established by sanitary survey. However, the bacterial ratio of atypical colonies to total coliform colonies (AC/TC), obtained from the total coliform membrane filter assay on m-Endo media, correctly indexed human faecal impact of inadequately sewered villages located along the creek. In addition, the AC/TC ratio correctly classified the predominant source of faecal runoff in the creek headwaters as agricultural, and indicated when aged agricultural faecal material was introduced by tributaries. An approach for watershed management that uses the AC/TC ratio in addition to levels of bacteria is proposed.     

Bacteriological eutrophication indicators in four Colombian water bodies (South America)

For the first time in Colombia the relative abundance of Pseudomonas spp., Vibrio spp., Plesiomonas shigelloides , the Aeromonas hydrophila group, and the faecal and total coliforms were studied in three dams and a natural lagoon with a different trophic state. Coliforms were low in all the water bodies, but their number was greater in the dams with a greater anthropic activity. Vibrio spp. was not detected. The density of P. shigelloides was irregular. Pseudomonas spp. was always present and showed relatively high abundance. The bacteria of the A. hydrophila group were constant and their numbers increased with the trophic state, for which they could be indicators of the eutrophication phenomena. Significant correlation ( P < 0.05) was established between total and faecal coliforms and pH, transparency, nitrites, nitrates, and orthophosphates. P. shigelloides was correlated with faecal coliforms and Aeromonas hydrophila with total coliforms and transparency.     

Bacteriological eutrophication indicators in four Colombian water bodies (South America)

For the first time in Colombia the relative abundance of Pseudomonas spp., Vibrio spp., Plesiomonas shigelloides , the Aeromonas hydrophila group, and the faecal and total coliforms were studied in three dams and a natural lagoon with a different trophic state. Coliforms were low in all the water bodies, but their number was greater in the dams with a greater anthropic activity. Vibrio spp. was not detected. The density of P. shigelloides was irregular. Pseudomonas spp. was always present and showed relatively high abundance. The bacteria of the A. hydrophila group were constant and their numbers increased with the trophic state, for which they could be indicators of the eutrophication phenomena. Significant correlation ( P < 0.05) was established between total and faecal coliforms and pH, transparency, nitrites, nitrates, and orthophosphates. P. shigelloides was correlated with faecal coliforms and Aeromonas hydrophila with total coliforms and transparency.     

The impact of cyanobacteria on growth and death of opportunistic pathogenic bacteria

Climate change may cause increased microbial growth in water sources and more knowledge is required on how this may affect the hygienic water quality, i.e., whether increased occurrence of cyanobacteria and algae may stimulate the growth rate of opportunistic pathogenic bacteria. Laboratory experiments were performed to investigate if the presence of the cyanobacteria Anabanea lemmermannii and Microcystis aeruginosa affected the survival and growth rate of the opportunistic pathogenic bacteria Aeromonas hydrophila and Pseudomonas aeruginosa, and the faecal indicators Escherichia coli and coliforms. Cyanobacteria were cultured in bottles containing the nutrient-poor medium 02. Sewage, A. hydrophila or P. aeruginosa was added to cyanobacterial cultures and the bacterial growth and survival was followed. E. coli and coliforms from sewage died within few days and the decay rate was not affected by the presence of cyanobacteria. The presence of Anabaena stimulated the growth rate of P. aeruginosa, but had no effect on the growth rate of A. hydrophila. Microcystis had no effect on the growth rate of P. aeruginosa and an inhibiting effect on the growth rate of A. hydrophila.     

Modelling faecal coliforms dynamics in the Seine estuary, France.

A model describing the dynamics of faecal coliforms (FC) in the Seine estuary has been developed and coupled with a three-dimensional hydrodynamic model. As input to this model three types of FC sources were considered: (1) FC transported by the Seine river flow at the estuary entrance at Poses dam; (2) FC brought in through the tributaries of the Seine estuary; and (3) the outfalls of the treated effluents of the WWTPs located along the estuary. As previous studies in the Seine estuary showed that a large proportion of FC was attached to SM, two pools of FC were considered separately in the model: free FC and FC attached to SM. Two main processes controlling the fate of FC in the estuary were considered: mortality and settling. The model calculates for a given discharge of the Seine river at Poses the longitudinal distribution of FC along the estuary. The model was validated by comparison of model calculations with experimental data in various hydrological conditions. The model also correctly reproduced the impact of the main river flow rate on the level of estuarine water's contamination. Finally, the model was used to test different scenarios of sanitation, suggesting priorities for sanitation efforts.     

Differentiation of faecal Escherichia coli from human and animal sources by random amplified polymorphic DNA-PCR (RAPD-PCR).

In this study the assessment of randomly amplified polymorphic DNA (RAPD) analysis was established as a molecular epidemiological tool. RAPD analysis was performed to differentiate faecal Escherichia coli isolates from human and animal sources. E. coli strains (128) were isolated from human and animal faeces (from cattle and sheep). Genomic DNA was extracted and randomly amplified polymorphic DNA-PCR (RAPD-PCR) fingerprinting was performed. Seven arbitrary primers were tested with a view to discriminating between E. coli isolates from humans and E. coli isolates from animals. RAPD profiles were analysed with hierarchical cluster analysis using an unweighted pair group method. RAPD profiles obtained with three of the tested primers (1247, 1290 and 1254) established a distinct differentiation between E. coli isolates from humans and E. coli from animals. Low levels of misclassification and high levels of specificity make RAPD a sensitive, efficient and reliable means of distinguishing closely related strains.     

Bacterial indicators of faecal pollution in the waters of the El-Kabir River and Akkar watershed in Syria and Lebanon

Water samples for bacterial analyses were taken, when possible, at 41 sites in Lebanon and Syria in the El‐Kabir River watershed. Samples were analysed for total coliform (TC), faecal coliform (FC) and faecal streptococcus. The Lebanese samples also were analysed for the presence of sulphate‐reducing bacteria. The TC and FC concentrations were extremely high throughout the watershed, rendering the water unfit for any human uses. The origin of the bacteria is the untreated sewage waste from the numerous settlements situated throughout the watershed. This primary origin is supported by high ratios of FC/faecal streptococcus, although impacts from animal wastes also were observed. The bacterial levels were higher in summer than in winter, indicating a high winter river flow that dilutes a relatively constant source of human sewage discharged to the system. Spring waters also exhibited elevated levels of bacteria, implicating surface land use and waste disposal practices upstream of the springs. The data clearly shows that human health is at a high risk and that strategies for improving water quality and protecting spring water must be expeditiously implemented.     

Experience with the antibiotic resistance analysis and DNA fingerprinting in tracking faecal pollution at two lake beaches.

Posting or closing of swimming beaches because of faecal contamination is a widespread problem reported in many locations. In a risk-based approach to this problem, the risk to swimmers' health is assessed by field monitoring of indicator bacteria and the associated risks are managed by source controls and other remedial measures. In risk assessment, great advances have been made in recent years with the introduction of microbial source tracking (MST) techniques. Two such techniques, antibiotic resistance analysis and DNA fingerprinting, were applied in a study of causes of faecal contamination at two lake beaches in Toronto, Ontario. Both methods identified bird faeces as the dominant sources of E. coli. Coping with this type of pollution presents a major environmental challenge.     

Microbiological quality of reclaimed water used for golf courses' irrigation.

Microbial quality of reclaimed water used for irrigation in two golf courses located in the southern Iberian Peninsula (Spain and Portugal) was evaluated. Bacterial indicators for faecal pollution (total and faecal coliforms, Escherichia coli and enterococci) were tested by membrane filtration using appropriate selective media. In addition, somatic E. coli bacteriophages, enteric viruses (entero-, hepatitis A and rota-) and Legionella pneumophila were also analysed. The results obtained showed that all wastewater treatment processes reduced adequately the number of indicator microorganisms although a significant correlation between pathogenic and indicator microorganisms tested was not found. L. pneumophila was detected by PCR but not confirmed by culture. Survival experiments of pathogenic microorganisms in aerosols and irrigated turf are conducted to determine the health hazards for the golf practice and to propose a microbial standard for wastewater used for irrigation of golf courses.     

Subsurface constructed wetlands for wastewater treatment and reuse in agriculture: five years of experiences in Sicily, Italy.

The paper reports the results of an investigation on a H-SSF constructed wetland for tertiary treatment in a small community located in eastern Sicily, Italy. The system is designed to reuse wastewater for the irrigation of olive orchards covering about 150 hectares. From March 2001 to September 2005, wastewater samples were collected and the following chemical-physical parameters were determined according to Standard Methods: temperature, pH, EC, DO, TSS (105degrees C), BOD5, COD, TN, TP. The following microbiological parameters were also evaluated: total coliforms, faecal coliforms, Escherichia coli, faecal streptococci, Salmonella, helminth eggs. Mean removal efficiencies ranged from 77 to 92% (TSS), 37 to 72% (BOD5), 51 to 79% (COD), 97 to 99.5% (E. coli). Removal efficiency of Salmonella and helminth eggs was always 100% in all samples; however, only 150% of total samples matched the limit of E. coli (< 50 CFU/100 mL) fixed by Italian legislation for wastewater reuse.     

Direct detection of bacterial faecal indicators in water samples using PCR.

The presence of enteric pathogens in water resources represents a serious risk for public health. Therefore, their precise detection, and especially detection of E. coli, which is obviously regarded as the main indicator of faecal contamination of water, is an essential step in ensuring bacterial safety of water. Numerous PCR protocols for detection of E. coli have been published to date. They are usually based on amplification of regions derived from lacZ (beta-D-galactosidase) and uidA (beta-D-glucuronidase) gene sequences. However, these methods are not universal enough for precise detection of all E. coli strains found in water samples. We developed a novel triplex PCR method for detection of E. coli in which cyd gene coding for cytochrome bd complex was co-amplified along with lacZ and uidA genes. Our triplex PCR approach significantly increases the specificity and reliability of E. coli detection in water samples. This approach allowed us to distinguish Shigella flexneri from E. coli. In addition, we were able to detect even non-coliform Klebsiella and Raoutella spp., some of which can also cause infections to humans.     

Predictive input parameters for enteric virus presence at the inlet of a potable water supply.

Kentucky river water samples were collected at the input of a potable water supply weekly, for a period of two years. Eighteen parameters were analysed and/or recorded for each weekly observation. The variables were of a physical, chemical and biological nature. Enteric virus presence was detected by cell culture analysis from approximately 200 L samples of river water. A database of 108 observations was assembled. Of these only 100 complete observations were used for the modelling analysis. Model input parameters were selected solely based on their statistical significance in forward, backward and stepwise selection procedures. Multivariate logistic regression (MLR) was used to determine the relative significance of the input parameters for predicting the presence or absence of enteric virus. The final, best-fit MLR model that used log-transformed values for enterococci, the AC/TC ratio, faecal coliforms (FC) and temperature demonstrated 82.5 and 81.4% sensitivity and specificity, respectively.     

Assessment of faecal pollution and relative algal abundances in Lakes Oconee and Sinclair, Georgia, USA

Water quality was assessed for five subregions of Lake Oconee and Lake Sinclair in Georgia, USA. The areas were chosen for their levels of human impact, including: (i) suburban development >30 years old; (ii) modern suburban development; (iii) industry; (iv) agriculture; and (v) an area of low human activity. The measured temperature, pH, dissolved oxygen concentration and turbidity values were normal for oligotrophic lakes. Faecal pollution in the lakes was determined using the membrane filtration method (Method 1600) on mEI plates for enterococci. There was a positive correlation between turbidity and the number of faecal enterococci in the lakes. The faecal pollution level was higher for the old suburban and agricultural areas. Faecal pollution in the agricultural region of Lake Oconee exceeded EPA regulatory standards. The faecal pollution source was identified using polymerase chain reaction detection, with Bifidobacterium adolescentis being a marker of human faecal pollution, and bovine-associated Bacteroides (BoBac) as a marker of cattle faecal pollution. Human faecal pollution was detected in the agricultural, old suburban and industrial areas of the lakes. In contrast, bovine faecal pollution was detected only in the agricultural area of Lake Oconee. Measurements of chlorophyll-a and relative algal community abundance indicated the least-impacted and modern suburban areas had significantly lower numbers of primary producers, being dominated by diatoms.     

Indigenous soil bacteria and low moisture may limit but allow faecal bacteria to multiply and become a minor population in tropical soils.

The soil environment in Hawaii is generally characterised as sub-optimal but permissive to support the in situ growth of E. coli and enterococci. However, soil desiccation and competition for nutrients by major indigenous soil microflora have been identified as potential factors that could limit a rapid and continual growth of faecal indicator bacteria in this soil environment. Despite these limitations, the genetic capacities of E. coli and enterococci are robust enough to enable these bacteria to become established as minor populations of Hawaii's soil microflora. Although the concentrations of E. coli and enterococci may have represented a fraction of the total soil microbiota, their presence in this habitat was very significant, for two important reasons: (a) soil was a major environmental source of E. coli and enterococci, and (b) the elevated counts of these bacteria in streams that routinely exceeded the EPA standards were due to run-off from soil. As a result, E. coli and enterococci were inadequate indicators to measure the degree of faecal contamination and potential presence of sewage-borne pathogens in Hawaiian streams.     

Microbiological quality of groundwater sources used by rural communities in Limpopo Province, South Africa.

A survey of the microbiological quality of water from 194 boreholes (97 privately owned and 97 communal boreholes) in the rural Thitale-Hlanganani area of the Limpopo Province, South Africa was carried out between August 2002 and August 2003. Very little information on the microbiological quality of privately-owned boreholes in rural communities is available raising concerns about the safety of these groundwater supplies. In this study, levels of total coliforms, thermotolerant (faecal) coliforms, faecal enterococci, Clostridium perfringens (vegetative cells and spores) and somatic coliphages were determined for community and privately-owned borehole water. The average counts for total coliforms, faecal coliforms, faecal enterococci and Clostridium perfringens exceeded the South African recommended guideline limits of 0-10 counts.100 ml(-1) for total coliforms and 0 counts. 100 ml(-1) for faecal coliforms, faecal enterococci and Clostridium perfringens respectively. Comparisons between the levels of indicator bacteria present in private and communal boreholes during dry seasons indicated a statistical difference for faecal enterococci bacteria (p = 0.005) and Clostridium perfringens (p = 0.08). Comparisons between the levels of indicator bacteria present in private and communal boreholes during rainy seasons indicated statistical differences between total coliforms (p = 0.05), faecal coliforms (p = 0.03) and Clostridium perfringens (p = 0.009) bacteria. No significant differences in the presence of somatic coliphages in both private and communal borehole water were see. The results indicated the need for environmental impact assessment studies to monitor the microbiological quality of groundwater sources in rural communities.     

Impact of Treated Wastewater Reuse on Agriculture and Aquifer Recharge in a Coastal Area: Korba Case Study

Treated wastewater (TWW) reuse has increasingly been integrated in the planning and development of water resources in Tunisia. The present study aimed the evaluation of the environmental and health impact that would have the reuse of TWW for crops direct irrigation or for the recharge of the local aquifer in Korba (Tunisia). For this purpose water analyses were carried on the TWW intended for the aquifer recharge and on underground water of this area. As for underground water before recharge, no contamination by organic matter or heavy metals is shown but high salinity, nitrate, potassium and chloride concentrations are detected. The bacteriological analyses show the occurrence of faecal streptococcus, thermo-tolerant coliforms, total coliforms and E coli, but absence of salmonella. These results indicate that this water is not suitable for irrigation worse still for drinking purpose. The monitoring of TWW pollutants has demonstrated that oxygen demands (COD and BOD) do not exceed the Tunisian standards for TWW used in agriculture (NT 106.03) except for August when samples reach high values (COD = 139 mg O₂ L^− 1, BOD = 34). It is also the case for temperature, electrical conductivity (EC), salinity and pH. Heavy metal concentrations are under the detection limit. The determination of nutrients shows relatively low concentrations of nitrates, nitrites and orthophosphate (the maxima in mg L^− 1 are respectively 6.6, 5.6 and 0.92) whereas the potassium levels are high (up to 48.8 mg L^− 1) and the ammonia levels very high, reaching 60.6 mg L^− 1. As for bacteriological pollution, while no salmonella and intestinal nematods are detected, high concentrations of total coliforms, thermo-tolerant coliforms, faecal streptococci and E. coli are analysed. Consequently, the better use of TWW in this region would be the use of infiltration basins for the recharge of the deteriorated aquifer by TWW. It would give the opportunity to better the quality of the TWW reaching the groundwater by an additional treatment for bacteriological and suspended solid (TSS) contaminants while being an alternative water for the aquifer recharge and a coastal barrier against seawater intrusion.     

Sources and Sinks of Escherichia coli in Benthic and Pelagic Fish

Escherichia coli and fecal coliform bacteria were isolated from five benthic and four pelagic fish species to determine their role in the fecal contamination of recreational waters. All fish were collected during fall 2006 from Southworth Marsh in the Duluth-Superior Harbor, a public beach that is commonly posted to minimize water contact due to high E. coli levels. Although fecal coliform bacteria were isolated from each fish species, they were only isolated from 66% and 72% of the individual benthic and pelagic fish, respectively. While 42% of the fecal coliforms from benthic fish were E. coli, only 4% of these bacteria from pelagic fish were E. coli. Cluster analysis showed different fish species harbored identical strains of E. coli and some fish contained multiple E. coli strains. The potential source for 65% of the E. coli isolates obtained from fish were identified by using the HFERP DNA fingerprinting method and libraries of E. coli DNA fingerprints from warm-blooded animals and environmental isolates collected in the area. The E. coli strains whose source could be identified were most similar to strains isolated from sediments, Canada geese, mallard ducks, and wastewater. None of the fish E. coli had DNA fingerprints matching those from any water or beach sand isolates. Although our results demonstrate that benthic fish contain E. coli, it may be more appropriate to consider these fish as a vector of E. coli from other sources, rather than a new source of E. coli contamination in aquatic environments.     

不同水生植物对大肠杆菌去除作用的比较研究

目前国内外水体污染防治针对病原菌的去除技术研究甚少,基于研究区域内已有的人工湿地和生态沟渠基本配置形式,通过室内常温盆栽试验筛选可高效去除病原菌(总大肠菌群和埃希氏大肠杆菌)的水生植物,并在自然生态沟渠(200 m)进行实地筛选。结果表明:水生植物能通过根系分泌物(降低水体pH)抑制大肠杆菌生存繁殖,并增加湿地拦截吸附能力;大部分水生植物组能有效吸附并去除埃希氏大肠杆菌,其中以水芹去除效果最佳;除香菇草、野茭白外,其他水生植物均能有效去除总大肠菌群;自然生态沟渠中再力花、美人蕉、岩菖蒲对总大肠菌群去除效率较高。总体而言,岩菖蒲、狐尾藻、再力花、水葫芦、美人蕉、水芹对大肠杆菌的去除率较高,可作为适宜的生态沟渠水生植物用于生态防治实践中。