The limitation of the ratio of fecal coliforms to total coliphage as a water pollution index

The fecal coliform populations of raw sewage, sewage lagoon effluent, and river water were determined using the most probable number technique. The total coliphage populations of the three water sources were determined using Escherichia coli B (ATCC 11303-1) host cells. The ratios of fecal coliforms to coliphage in the three water samples were 87:1, 4.2:1, and 0.15:1, respectively. The ratio of fecal coliforms to coliphage in stored raw sewage decreased from 87:1 to about 1:1 within 7 days at 20°C and within 28 days at 4°C. These changing ratios resulted from the greater longevity of the coliphage compared with that of the coliform bacteria. The use of the ratio of fecal coliforms to coliphage is not considered reliable as an index of when a fecal pollution event occurred because the ratio is influenced by prior contamination, presence of sediment, chlorination, and temperature.     

Impact of population and latrines on fecal contamination of ponds in rural Bangladesh

A majority of households in Bangladesh rely on pond water for hygiene. Exposure to pond water fecal contamination could therefore still contribute to diarrheal disease despite the installation of numerous tubewells for drinking. The objectives of this study are to determine the predominant sources (human or livestock) of fecal pollution in ponds and examine the association between local population, latrine density, latrine quality and concentrations of fecal bacteria and pathogens in pond water. Forty-three ponds were analyzed for E. coli using culture-based methods and E. coli, Bacteroidales and adenovirus using quantitative PCR. Population and sanitation spatial data were collected and measured against pond fecal contamination. Humans were the dominant source of fecal contamination in 79% of the ponds according to Bacteroidales measurements. Ponds directly receiving latrine effluent had the highest concentrations of fecal indicator bacteria (up to 10⁶ Most Probable Number (MPN) of culturable E. coli per 100 mL). Concentrations of fecal indicator bacteria correlated with population surveyed within a distance of 30-70 m (p < 0.05) and total latrines surveyed within 50-70 m (p < 0.05). Unsanitary latrines (visible effluent or open pits) within the pond drainage basin were also significantly correlated to fecal indicator concentrations (p < 0.05). Water in the vast majority of the surveyed ponds contained unsafe levels of fecal contamination attributable primarily to unsanitary latrines, and to lesser extent, to sanitary latrines and cattle. Since the majority of fecal pollution is derived from human waste, continued use of pond water could help explain the persistence of diarrheal disease in rural South Asia.     

Peak flux performance and microbial removal by selected membrane bioreactor systems

A pilot study was conducted over a period of 18 months at the Point Loma Wastewater Treatment Plant (PLWWTP) in San Diego, CA to evaluate the operational and water quality performance of six selected membrane bioreactor (MBR) systems at average and peak flux operation. Each of these systems was operated at peak flux for 4 h a day for six consecutive days to assess peak flux performance. Virus seeding studies were also conducted during peak flux operation to assess the capability of these systems to reject MS-2 coliphage. When operating at steady state, these MBR systems achieved an effluent BOD concentration of <2 mg/L and a turbidity of <0.1 NTU. Peak flux for the MBR systems ranged from 56 to 76 L/m²/h (liters per square meter per hour) with peaking factors in the range of 1.5-3.2. When switching from average to peak flux operation, a reversible drop of 22-32% in temperature-corrected permeability was observed for all submerged MBR systems. The percent drop in permeability increased as MLSS concentration in the membrane tank increased from 11,100 mg/L to 15,300 mg/L and was observed to be highest for the system operating at highest MLSS concentration. Such trends were not observed with an external MBR system. Each MBR system was able to sustain a 4-h-a-day peak flow for six consecutive days with only moderate membrane fouling. The membrane fouling was quantified by measuring the drop in temperature-corrected permeability. This drop ranged from 13 to 33% over six days for different submerged MBR systems. The MBR systems achieved microbial removal in the range of 5.8-6.9 logs for total coliform bacteria, >5.5 to >6.0 logs for fecal coliform bacteria and 2.6 to >3.4 logs for indigenous MS-2 coliphages. When operating at peak flux, seeded MS-2 coliphage removal ranged from 1.0 to 4.4 logs, respectively. The higher log removal values (LRVs) for indigenous MS-2 coliphage among different MBR systems were probably the result of particle association of indigenous coliphage. Differences in membrane pore size (0.04-0.2 μm) amongst the MBR systems evaluated did not have a substantial impact on indigenous MS-2 coliphage removal, but seeded MS-2 coliphage removal varied among the different MBR systems.     

Isolation and characterization of Bacteroides host strain HB-73 used to detect sewage specific phages in Hawaii

Previous studies have shown that Escherichia coli and enterococci are unreliable indicators of fecal contamination in Hawaii because of their ability to multiply in environmental soils. In this study, the method of detecting Bacteroides phages as specific markers of sewage contamination in Hawaii’s recreational waters was evaluated because these sewage specific phages cannot multiply under environmental conditions. Bacteroides hosts (GB-124, GA-17), were recovered from sewage samples in Europe and were reported to be effective in detecting phages from sewage samples obtained in certain geographical areas. However, GB-124 and GA-17 hosts were ineffective in detecting phages from sewage samples obtained in Hawaii. Bacteroides host HB-73 was isolated from a sewage sample in Hawaii, confirmed as a Bacteroides sp. and shown to recover phages from multiple sources of sewage produced in Hawaii at high concentrations (5.2-7.3 × 10⁵ PFU/100 mL). These Bacteroides phages were considered as potential markers of sewage because they also survived for three days in fresh stream water and two days in marine water. Water samples from Hawaii’s coastal swimming beaches and harbors, which were known to be contaminated with discharges from streams, were shown to contain moderate (20-187 CFU/100 mL) to elevated (173-816 CFU/100 mL) concentrations of enterococci. These same samples contained undetectable levels (<10 PFU/100 mL) of F+ coliphage and Bacteroides phages and provided evidence to suggest that these enterococci may not necessarily be associated with the presence of raw sewage. These results support previous conclusions that discharges from streams are the major sources of enterococci in coastal waters of Hawaii and the most likely source of these enterococci is from environmental soil rather than from sewage.     

Sulfamethoxazole contamination of a deep phreatic aquifer

Groundwater samples were obtained from the water table region of a phreatic aquifer (unsaturated zone depth up to 28 m) under land irrigated with wastewater effluents for about 5 decades and a relatively deep pumping well (109 m), used as a drinking water source till 2007, located downstream (1300 m) of wastewater effluent and sludge infiltration facilities. Sulfamethoxazole (SMX) concentrations in secondary effluents varied between 90 and 150 ng/L. SMX was extracted using SPE and was analyzed by HPLC-MS/MS. SMX (maximum concentration of 37 ng/L) was detected in the water table region, in two monitoring wells, after an unsaturated zone transport period of about 16 years. The maximum SMX concentration detected in the pumping well was of 20 ng/L. These results question wastewater effluent disposal strategies including the suitability of irrigation with effluents on the replenishment area of an aquifer supplying drinking water.     

Survival of prototype strains of somatic coliphage families in environmental waters and when exposed to UV low-pressure monochromatic radiation or heat

The potential use of specific somatic coliphage taxonomic groups as viral indicators based on their persistence and prevalence in water was investigated. Representative type strains of the 4 major somatic coliphage taxonomic groups were seeded into reagent water and an ambient surface water source of drinking water and the survival of the added phages was measured over 90 days at temperatures of 23-25 and 4 °C. Microviridae (type strain PhiX174), Siphoviridae (type strain Lambda), and Myoviridae (type strain T4) viruses were the most persistent in water at the temperatures tested. The Microviridae (type strain PhiX174) and the Siphoviridae (type strain Lambda) were the most resistant viruses to UV radiation and the Myoviridae (type strain T4) and the Microviridae (type strain PhiX174) were the most resistant viruses to heat. Based on their greater persistence in water over time and their relative resistance to heat and/or UV radiation, the Myoviridae (type strain T4), the Microviridae (type strain PhiX174), and the Siphoviridae (type strain Lambda) were the preferred candidate somatic coliphages as fecal indicator viruses in water, with the Microviridae (type strain PhiX174) the most resistant to these conditions overall.     

North and south american studies on the potential of coliphage as a water quality indicator

Studies were undertaken to assess the potential of coliphages to be used universally as water quality indicators and more specifically as health hazard indicators. Data were obtained from three water bodies, a northern Canadian River, inshore water samples from Lake Ontario and from marine beaches in Brazil. Data from this two continent, three water body study indicated (a) that within location fecal coliform and coliphages are positively correlated, (b) coliphage values can be predicted by using fecal coliform MPN, fecal streptococci MF and E. coli MF data and (c) a water quality guideline of 20 coliphage/100 ml for recreational fresh waters is proposed.     

Removal of native coliphages and coliform bacteria from municipal wastewater by various wastewater treatment processes: implications to water reuse

The efficacy of a conventional activated sludge wastewater treatment process and the membrane bioreactor technology in removing microbial pathogens was investigated. Total and fecal coliforms and somatic and F-specific coliphages were used as indicators of pathogenic bacteria and viruses. Up to 5.7 logs removal of coliforms and 5.5 logs of coliphages were observed in the conventional treatment process with advanced tertiary treatment. Addition of chemical coagulants seemed to improve the efficacy of primary and secondary treatment for microorganism removal. Complete removal of fecal coliforms and up to 5.8 logs removal of coliphages was observed in the MBR system. It was shown that the MBR system was capable of high removal of coliphages despite the variation in feed coliphage concentrations. The results of this study indicated that the MBR system can achieve better microbial removal in far fewer steps than the conventional activated sludge process with advanced tertiary treatment. The final effluent from either treatment processes can be potentially reused.     

F-specific RNA coliphages: occurrence,types, and survival in natural waters

A small, well-defined watershed was investigated over a 2-year period to determine the prevalence of F-specific RNA coliphage (F + RNA) serotypes as indicators of animal fecal contamination. Sampling sites collected runoff from areas of urban and agricultural land use patterns. F-specific coliphages were concentrated from 2-L freshwater samples by polyethylene glycol precipitation, isolated using the double agar layer (DAL) method, confirmed as F + RNA by RNAse suppression, and serotyped. A subset of serotyped F + RNA were confirmed by genotyping. To determine relative survival, 10 confirmed F + RNA field isolates and 5 prototypic F + RNA were spiked into surface water and incubated at 25 degrees C for 36 days. F-specific coliphage isolation was strongly associated with rainfall events and was infrequent from primarily animal impacted surface waters. Field isolates were predoffiinantly Type I F + RNA (81%) and raw sewage isolates were predominantly Type III F + RNA (57%). Genotyping from either the watershed or raw sewage samples never positively identified Type IV F + RNA. Results from laboratory studies showed that F + RNA differ in their survival in water and that Type IV strains were the least persistent. Type III F + RNA were found to be reliably related to the release of uncontrolled human fecal material in the watershed, but the results of this study suggest that further study is required before utilizing for fecal source identification in natural waters.     

Investigation of norovirus occurrence in groundwater in metropolitan Seoul, Korea

Groundwater is an important source of drinking and household water worldwide. Hence, the quality of groundwater is very important for preventing waterborne disease outbreaks and should be properly monitored. This study investigated the prevalence of waterborne viruses and fecal indicators in groundwater in metropolitan Seoul and Gyeonggi province, South Korea. A total of 116 samples of groundwater were taken using NanoCeram filters during both summer (June to August) and fall-winter seasons (October to December) in 2008. Among 71 sampling sites, 28 (48.3%) and 18 (35.3%) were positive for norovirus (NoV) from the summer and fall-winter season, respectively. The identified genotypes of NoV include GI-1, 4, 8, 9 and GII-4, 10, 11 (or 17), 13, 15 (or 16). None of fecal indicators was significantly correlated with NoV in groundwater. Among the tested fecal indicators, somatic coliphage (95.3%) showed an excellent true-negative rate of NoV occurrence. The combination of chemical, microbial and viral indicators increased the positive predictive value (50-100%). This study demonstrated a high prevalence of NoV in groundwater in metropolitan Seoul areas and characterized the positive and negative predictive values of a fecal indicator for predicting NoV prevalence.     

Fate of physical,chemical, and microbial contaminants in domestic wastewater following treatment by small constructed wetlands

In order to evaluate the efficacy of constructed wetlands for treatment of domestic wastewater for small communities located in rural areas, small-scale wetland mesocosms (400 L each) containing two treatment designs (a mixture of Typha, Scirpus, and Juncus species; control without vegetation) were planted into two depths (45 or 60 cm) with pea gravel. Each mesocosm received 19 L/day of primary-treated domestic sewage. Mesocosms were monitored (inflow and outflow samples) on a monthly basis over a 2-year period for pH, total suspended solids (TSS), 5-day biochemical oxygen demand (BOD(5)), total Kjeldahl nitrogen (TKN), dissolved oxygen (DO), and conductivity. Microbiological analyses included enumeration of fecal coliforms, enterococci, Salmonella, Shigella, Yersinia, and coliphage. Significant differences between influent and effluent water quality for the vegetated wetlands (p<0.05) were observed in TSS, BOD(5), and TKN. Increased DO and reduction in fecal coliform, enterococcus, Salmonella, Shigella, Yersinia, and coliphage populations also were observed in vegetated wetlands. Greatest microbial reductions were observed in the planted mesocosms compared to those lacking vegetation. Despite marked reduction of several contaminants, wetland-treated effluents did not consistently meet final discharge limits for receiving bodies of water. Removal efficiencies for bacteria and several chemical parameters were more apparent during the initial year compared to the second year of operation, suggesting concern for long-term efficiency and stability of such wetlands.     

Distribution of enteroviruses in sediments contiguous with a deep marine sewage outfall

The distribution of enteroviruses, coliforms, fecal coliforms and fecal streptococcus discharged from a raw sewage outfall pipe 3.6 km from shore at a depth of 44 m was studied. Enteroviruses and indicator bacteria were only isolated from water in an area within 200 m of the outfall. However, viruses were isolated from sediments as far as 3.6 km from the outfall at recreational bathing beaches. The concentration of enteroviruses in the sediment at the outfall boil ranged from 112 to 78 PFU l⁻¹ and 0 to 30 PFU l⁻¹ at the bathing beach. Indicator bacteria were not detected in the water or sediment at the bathing beach. Analysis of sediments for viruses may give a better idea of the long term distribution of sewage discharged from the outfall pipe.     

Disinfection efficiency of peracetic acid, UV and ozone after enhanced primary treatment of municipal wastewater

The City of Montreal Wastewater Treatment Plant uses enhanced physicochemical processes (ferric and/or alum coagulation) for suspended solids and phosphorus removal. The objective of this study was to assess the ability of peracetic acid (PAA), UV, or ozone to inactivate the indicator organisms fecal coliforms, Enterococci, MS-2 coliphage, or Clostridium perfringens in the effluent from this plant. PAA doses to reach the target fecal coliform level of 9000 CFU/100mL exceeded 6 mg/L; similar results were obtained for enterococci, and no inactivation of Clostridium perfringens was observed. However a 1-log reduction of MS-2 occurred at PAA doses of 1.5 mg/L and higher. It was expected that this effluent would have a high ozone demand, and would require relatively high UV fluences, because of relatively high effluent COD, iron and suspended solids concentrations, and low UV transmittance. This was confirmed herein. For UV, the inactivation curve for fecal coliforms showed the typical two-stage shape, with the target of 1000 CFU/100 mL (to account for photoreactivation) occurring in the asymptote zone at fluences >20 mJ/cm(2). In contrast, inactivation curves for MS-2 and Clostridium perfringens were linear. Clostridium perfringens was the most resistant organism. For ozone, inactivation was already observed before any residuals could be measured. The transferred ozone doses to reach target fecal coliform levels ( approximately 2-log reduction) were 30-50 mg/L. MS-2 was less resistant, but Clostridium perfringens was more resistant than fecal coliforms. The different behaviour of the four indicator organisms studied, depending on the disinfectant, suggests that a single indicator organism might not be appropriate. The required dose of any of the disinfectants is unlikely to be economically viable, and upstream changes to the plant will be needed.     

Occurrence of perchlorate in drinking water sources of metropolitan area in Japan

The occurrence of perchlorate in the Tone River Basin was investigated using an ion chromatograph (IC) coupled with a tandem mass spectrometer (MS/MS). Perchlorate was found at high concentrations in the upper Tone River and its tributary, Usui River, and the maximum concentrations were 340 and 2300 microg/L, respectively. The possible sources of perchlorate in two areas were attributable to industrial effluents. In case of the upper Tone River, perchlorate concentration in an effluent was 1100 microg/L and its concentrations in a tributary (or waterway) directly downstream of the outlet of the effluent ranged from 44 to 1500 microg/L. In case of the Usui River, perchlorate concentration in another effluent was 15,000 microg/L and its concentrations downstream of the outlet of the effluent were 1100-3900 microg/L. Due to the discharge of perchlorate in the upper Tone River Basin, perchlorate concentrations in the river waters of the middle and lower Tone River Basin were generally 10-20 microg/L. Perchlorate concentrations in 30 tap water samples were investigated. Water sources of three tap water samples were other than the Tone River Basin and their perchlorate concentrations were 0.16-0.87 microg/L. On the other hand, water sources of the remaining 27 samples were the upper, middle and lower Tone River Basin and their perchlorate concentrations were 0.06-37 microg/L. Perchlorate concentrations were more than 1 microg/L in 19 tap water samples and more than 10 microg/L in 13 samples. It was shown that tap waters in the Tone River Basin were widely contaminated with perchlorate. To our knowledge, this study was the first to report on perchlorate contamination of environmental and drinking waters in Japan.     

Virus removal during simulated soil-aquifer treatment

Removals of indigenous coliphage and seeded poliovirus type 1 during simulated soil-aquifer treatment were evaluated during transport of secondary effluent under unsaturated flow conditions in 1-m soil columns. Independent variables included soil type (river sand or sandy loam) and infiltration rate. Removal of coliphage was in all cases less than removal of poliovirus type 1 (strain LSc-2ab), supporting contentions that indigenous coliphage can act as a conservative indicator of groundwater contamination by viral pathogens of human origin. Coliphage retention was significantly more efficient (p<0.001) in the finer-grained sandy loam (93%) than in sand (76%). Increasing reactor detention time from 5 to 20 h increased coliphage attenuation from 70% to 99% in a 1-m sand column. There was a significant linear correlation (p=0.012) between log-transformed (fractional) coliphage concentration [log(C/C(0))] and reactor detention time. Re-mobilization of attached coliphage occurred during simulated rainfall using low-ionic-strength water. Inhibition of aerobic respiration resulted in significantly less efficient coliphage attenuation (p=0.033), suggesting the involvement of aerobic microorganisms in the survival/retention of this virus.     

Monitoring source water for microbial contamination: evaluation of water quality measures

Watershed management programs often rely on monitoring for a large number of water quality parameters to define contaminant issues. While coliforms have traditionally been used to identify microbial contamination, these indicators cannot discriminate among potential contaminant sources. Microbial source tracking (MST) can provide the missing link that implicates the sources of contamination. The objective of this study was to use a weight-of-evidence approach (land use analysis using GIS, sanitary surveys, traditional water quality monitoring, and MST targets) to identify sources of pollution within a watershed that contains a raw drinking water source. For the study watersheds, statistical analyses demonstrated that one measure each of particulate matter (turbidity, particle counts), organic matter (total organic carbon, dissolved organic carbon, UV(254) absorbance), and indicator organisms (fecal coliforms, enterococci) were adequate for characterizing water quality. While these traditional parameters were useful for assessing overall water quality, they were not intended to differentiate between microbial sources at different locations. In contrast, the MST targets utilized (Rhodococcus coprophilus, sorbitol-fermenting Bifidobacteria, and male-specific coliphages) pinpointed specific sources of microbial pollution. However, these targets could not be used for routine monitoring due to a high percentage of non-detects.     

Hydrogeological investigation of ground water arsenic contamination in south Calcutta

Typical clinical symptoms of acute arsenic poisoning have been detected in 1000 residents near a factory in P.N. Mitra Lane, Behala, South Calcutta, located in a thickly populated area manufacturing copper acetoarsenite (Paris-Green) an arsenical pesticide for the past 25 years. Soil around the effluent dumping point of the factory was exceptionally contaminated, with arsenic, copper and chromium concentrations of 20,100-35,500 mg kg-1, 33,900-51,100 mg kg-1 and 5300-5510 mg kg-1. Arsenic and copper concentrations in bore-hole soils collected up to a depth of 24.4 m at the effluent dumping point, decreased with depth. Arsenous acid, arsenic acid, methylarsonic acid (MA) and dimethylarsinic acid (DMA) were detected in bore-hole soils up to a depth of 1.37 m, after which only inorganic arsenical compounds were present. A positive correlation was established between arsenic and copper authenticated the Paris-Green waste disposal site as the source of contamination. Mechanism of ground water contamination from this disposal site had been probed by a systematic hydrogeological survey and the arsenic content of the tube-well waters in the surrounding areas. Hydraulic conductivity was maximum in the central part. The site for disposal of the effluent was a ditch located in the zone of discharge. Sparingly soluble Paris-Green cumulatively deposited in the waste disposal site is decomposed by micro-organisms to water-soluble forms and finally percolated to underground aquifers along with rain water through the discharge zone. The contaminant is currently moving towards WNW with ground water flow and the residents in the direction of encroaching contamination are insecure due to penetration of the contaminant.     

Uptake and release of zinc by aquatic bryophytes (Fontinalis antipyretica L. ex. Hedw.)

The zinc uptake and posterior release by an aquatic bryophyte—Fontinalis antipyretica L. Ex Hedw.—was experimentally studied in laboratory exposing the plants to different zinc concentrations in the range, 1.0–5.0 mg l⁻¹, for a 144 h contamination period, and then exposed to metal-free water for a 120 h decontamination period. The experiments were carried out in perfectly mixed contactors at controlled illumination, using mosses picked out in February 1997, with a background initial zinc concentration of 263 mg g⁻¹ (dry wt.). A first-order mass transfer kinetic model was fitted to the experimental data to determine the uptake and release constants, k₁ and k₂, the zinc concentration in mosses at the end of the uptake period, C_mu, and at the equilibrium, for the contamination and decontamination stages, C_me and C_mr, respectively. A bioconcentration factor, BCF=k₁/k₂ (zinc concentration in the plant, dry wt./zinc concentration in the water) was determined. A biological elimination factor defined as BEF=1−C_mr/C_mu was also calculated. BCF decreases from about 4500 to 2950 as Zn concentration in water increases from 1.05 to 3.80 mg l⁻¹. BEF is approximately constant and equal to 0.80. Comparing Zn and Cu accumulation by Fontinalis antipyretica, it was concluded that the uptake rate for Zn (145 h⁻¹) is much lower than for Cu (628 h⁻¹) and the amount retained by the plant decreased by a factor of about seven.