Evaluating sewage-associated JCV and BKV polyomaviruses for sourcing human fecal pollution in a coastal river in Southeast Queensland, Australia

In this study, the host-sensitivity and host-specificity of JC virus (JCV) and BK virus (BKV) polyomaviruses were evaluated by testing wastewater and fecal samples from nine host groups in Southeast Queensland, Australia. The JCV and BKV polyomaviruses were detected in 63 human wastewater samples collected from primary and secondary effluent, suggesting high sensitivity of these viruses in human wastewater. In the 81 animal wastewater and fecal samples tested, 80 were polymerase chain reaction (PCR) negative for the JCV and BKV markers. Only one sample (out of 81 animal wastewater and fecal samples) from pig wastewater was positive. Nonetheless, the overall host-specificity of these viruses to differentiate between human and animal wastewater and fecal samples was 0.99. To our knowledge, this is the first study in Australia that reports on the high specificity of JCV and BKV polyomaviruses. To evaluate the field application of these viral markers for detecting human fecal pollution, 20 environmental samples were collected from a coastal river. In the 20 samples tested, 15% (3/20) and 70% (14/20) samples exceeded the regulatory guidelines for Escherichia coli and enterococci levels for marine waters. In all, five (25%) samples were PCR positive for JCV and BKV, indicating the presence of human fecal pollution in the coastal river investigated. The results suggest that JCV and BKV detection using PCR could be a useful tool for identifying human-sourced fecal pollution in coastal waters.     

Identifying sources of fecal contamination inexpensively with targeted sampling and bacterial source tracking

Most bacterial source tracking (BST) methods are too expensive for most communities to afford. We developed targeted sampling as a prelude to BST to reduce these costs. We combined targeted sampling with three inexpensive BST methods, Enterococcus speciation, detection of the esp gene, and fluorometry, to confirm the sources of fecal contamination to beaches on Georgia's Jekyll and Sea Islands during calm and stormy weather conditions. For Jekyll Island, the most likely source of contamination was bird feces because the percentage of Ent. faecalis was high (30%) and the esp gene was not detected. For the Sea Island beach during calm conditions, the most likely sources of fecal contamination were leaking sewer lines and wildlife feces. The leaking sewer lines were confirmed with fluorometry and detection of the esp gene. For the Sea Island beach during stormflow conditions, the most likely sources of fecal contamination were wildlife feces and runoff discharging from two county-maintained pipes. For the pipes, the most likely source of contamination was bird feces because the percentage of Ent. faecalis was high (30%) and the esp gene was not detected. Sediments were also a reservoir of fecal enterococci for both Jekyll and Sea Islands. Combining targeted sampling with two or more BST methods identified sources of fecal contamination quickly, easily, and inexpensively. This combination was the first time targeted sampling was conducted during stormy conditions, and the first time targeted sampling was combined with enterococcal speciation, detection of the esp gene, and fluorometry.     

Monitoring of natural and synthetic hormones in a polluted river

Natural (estradiol, estrone, testosterone, estriol) and synthetic hormones (ethinylestradiol) are constantly excreted into the environment from human and animal sources but little is known of their transport. The purpose of this study was to determine how far along a 100 km river course that hormones could be detected after contamination with sewage effluent or fishpond effluent. Fourteen sites in the Lower Jordan River drainage were sampled (two sites above the sewage effluent contamination, eight sites below the contamination and four tributaries) before and after the dry season of 2002 (Spring and Fall). Samples were tested for testosterone, estrogen (estrone and estradiol combined), estriol, ethinylestradiol, ammonia and fecal coli. It was found that the fecal coli count dropped exponentially (from 250,000 to 60/100 ml3) and the ammonia dropped from 15 to less than 1mg/l over the initial 25 km stretch. Over the same stretch, the hormone values declined by half from their maximum values for testosterone (3.3 ng/l), estriol (8.8 ng/l), ethinylestradiol (6.1 ng/l), and estrogen (4.9 ng/l). From 67 to 100 km mark, testosterone (4.8 ng/l) and estrogen (2.4 ng/l) were still elevated while ethinylestradiol and estriol were >or=1.5 ng/l. The high level of testosterone and estrogen between 67 and 100 km marks was probably due to major discharge from fishponds between 23 and 27 km marks. Levels of ethinylestradiol above 1 ng/l, a level which can affect fish, was seen in 70% (12/16) of the samples tested. The data suggest that hormones in readily measured quantities can be transported considerable distances from the source of pollution.     

Estrogenic activity in the environment: municipal wastewater effluent, river, ponds, and wetlands

Estrogenic activity of regional water samples was evaluated. Samples obtained from wetlands and ponds involved in various agricultural land uses, from three river sites over four seasons, and from municipal wastewater effluent held in storage lagoons were evaluated. The estrogen-responsive cell line MCF-7 BOS was used in the E-screen assay to determine 17beta-estradiol equivalents (E2 Eq) of water samples extracted by solid-phase extraction. Estrogenic activity in surrounding wetlands and ponds from different land uses was not different, with 10(-12) M E2 Eq (0.3 ppt). Estrogenic activity of Red River samples was within the same range as wetland-pond samples. The highest activity was found downstream from municipal wastewater treatment effluent discharge sites, in winter when river flow was lowest (approximately 6 x 10(-13) M E2 Eq). Results showed that 7 of 20 wetland-pond samples and 5 of 12 river samples were below the limits of quantitation (approximately 3 x 10(-14) M E2 Eq). Toxicity was found in fall and summer river samples upstream from municipal wastewater release sites. The timing of toxicity did not coincide to the presence of elevated fecal coliforms. Estrogenic activity in wastewater effluent from lagoons decreased over time (approximately 25 to 5 x 10(-13) M E2 Eq) with an apparent half-life of 8 d for one lagoon. The median concentration of detectable estrogenic activity in regional water samples was approximately 50-fold less than the median 17beta-estradiol concentration of estradiol detected in some U.S. streams in previous studies.     

多管发酵法与其改进法检测水中粪大肠菌群的比较

粪大肠菌群是水体中粪便污染的重要指示菌,对于评价水质污染程度具有重要的卫生学意义。多管发酵法是传统的粪大肠菌群检测方法,但其步骤繁杂、周期长。目前,已有部分监测及科研人员对其进行了改进,笔者采用改进法与国标多管发酵法进行比较验证,认为Ec培养法可完全替代国标多管发酵法,纸片法亦可用于除酸性水样及菌量大的原污水外的其他环境水样中粪大肠菌群的检测。     

Antibiotic resistance profiles to determine sources of fecal contamination in a rural Virginia watershed

Antibiotic resistance analysis (ARA) was used to determine if enterococci of human origin were present in a stream (Spout Run) that passes through a rural nonsewered community (Millwood, VA). Millwood consists of 82 homes, all served by individual septic systems, and Spout Run drains a 5,800-ha karst topography watershed that contains large populations of livestock and wildlife. Periodic monitoring by state regulatory officials had resulted in Spout Run being placed on the Virginia impaired stream list and Millwood being categorized as an at-risk community. Stream samples were collected monthly and analyzed for fecal coliforms and enterococci (May 1999-May 2000); ARA was performed on enterococci stream isolates on a quarterly basis. All 117 stream samples were positive for fecal coliforms, and 32% exceeded the Virginia recreational water standard (1,000 fecal coliforms/100 mL). A library of 1,174 known source Enterococcus isolate antibiotic resistance profiles was constructed, and yielded correct classification rates of 94.6% for 203 human isolates, 93.7% for 734 livestock isolates, and 87.8% for 237 wildlife isolates. Antibiotic resistance analysis of 2,012 enterococcal isolates recovered from stream samples indicated isolates of human origin appeared throughout the stream as it passed through Millwood, with a yearly average of approximately 10% human, 40% wildlife, and 50% livestock. There were no human origin isolates in samples collected upstream from Millwood, and the percent human origin isolates declined downstream from Millwood. While a human signature was found in Spout Run, it was small compared with the proportion of isolates from livestock and wildlife.     

Cholesterol,beta-sitosterol,ergosterol, and coprostanol in agricultural soils

In this work we analyzed the sterol content of agricultural soils. Three eukaryotic sterols, cholesterol, beta-sitosterol, and ergosterol were chosen as representative of the animal, plant, and fungal kingdoms, while coprostanol was validated as a marker of human fecal matter contamination. Three soils subjected to different treatments (sewage sludge application, irrigation by saline waters, and contamination by industrial and municipal wastes) were sampled and their sterol content was measured and compared with adjacent untreated soils. The effects of time, location, and treatment were evaluated by means of a number of statistical techniques. Beta-sitosterol concentration varied from 0.9 to 30 mg kg(-1). Lesser values were measured in Cremona (2.1 mg kg(-1)) than in Bari (4.0 mg kg(-1)) and Naples (10.9 mg kg(-1)) soils. No significant effects were detected for cholesterol and ergosterol. Coprostanol was present after sewage sludge disposal and contamination by industrial and municipal wastes, while it was absent in the soil treated with saline water and in the adjacent untreated soil. Coprostanol concentration did not vary much within site and time of sampling, with a mean value of 0.2 mg kg(-1). We confirmed coprostanol as a useful persistent marker of human fecal matter contamination. Multivariate analysis highlighted a clear distinction between the eukaryotic sterols and coprostanol. In addition, a different behavior between ergosterol and cholesterol on one side and beta-sitosterol on the other was detected. This preliminary work suggests that sterols deserve a deeper study of their use as indicators in agricultural soils.     

MICROBIOLOGICAL QUALITY OF PUGET SOUND BASIN STREAMS AND IDENTIFICATION OF CONTAMINANT SOURCES1

ABSTRACT: Fecal coliforms, Escherichia coli, enterococci, and somatic coliphages were detected in samples from 31 sites on streams draining urban and agricultural regions of the Puget Sound Basin Lowlands. Densities of bacteria in 48 and 71 percent of the samples exceeded U.S. Environmental Protection Agency's freshwater recreation criteria for Escherichia coli and enterococci, respectively, and 81 percent exceeded Washington State fecal coliform standards. Male‐specific coliphages were detected in samples from 15 sites. Male‐specific F⁺RNA coliphages isolated from samples taken at South Fork Thornton and Longfellow Creeks were serotyped as Group II, implicating humans as potential contaminant sources. These two sites are located in residential, urban areas. F⁺RNA coliphages in samples from 10 other sites, mostly in agricultural or rural areas, were serotyped as Group I, implicating non‐human animals as likely sources. Chemicals common to wastewater, including fecal sterols, were detected in samples from several urban streams, and also implicate humans, at least in part, as possible sources of fecal bacteria and viruses to the streams.     

Environmental load of Cryptosporidium parvum oocysts from cattle manure in feedlots from the central and western United States

The first step in assessing the risk of water contamination by Cryptosporidium parvum oocysts from feedlot cattle (Bos taurus) production systems is to quantify the number of C. parvum oocysts present in the fecal material deposited by feedlot cattle. Our primary objective for this project was to estimate the daily environmental load of C. parvum oocysts in fecal material deposited by feedlot cattle from across the central and western USA. Our secondary goal was to genotype isolates of C. parvum from feedlot cattle to help facilitate proper identification of mammalian sources of waterborne C. parvum. Based on 5274 fecal samples from 22 feedlots in seven states (California, Washington, Colorado, Oklahoma, Texas, Nebraska, and South Dakota), we estimated a point prevalence of C. parvum of 0.99 to 1.08% in fecal material from feedlot pens from a wide range of climates and a diverse range of feedlot management systems. On average, fresh fecal material from throughout feedlot systems (recent arrivals to nearing slaughter) contained about 1.3 to 3.6 oocysts/g feces, which roughly translates to about 2.8 x 10(4) to 1.4 x 10(5) oocysts/animal per day.     

工业园区水污染控制多目标规划研究

本文分析了我国水污染控制规划的新特点，结合传统的水污染控制规划模型提出了适应新特点的流域工业园区水污染控制多目标规划的建模思路。新模型以污水排放口、污水处理厂、输水管线以及接纳污水的水体组成的水环境系统为基础。并纳入工业园区水污染治理子系统，同时考虑工业园区总量控制要求。     

CASS＋人工湿地工艺处理城镇污水工程实例

本文论述了成都市某小流域末端污水处理站的工艺流程,介绍了主要的构筑物和设备及其参数。该污水处理站属于成都市2009年小流域治污工程,采用CASS＋人工湿地工艺处理城镇污水。运行稳定后,且在进水稳定下出水氨氮和CODCr达到《城镇污水处理站污染物排放标准》（GB18918-2002）中的一级A类标准。     

Studies on the effect of Bladex and Etazine on bacterial indices of pollution

The effect of the triazine herbicides Etazine and Bladex on coliform and fecal streptococci groups was studied in Nile river water, Nile water seeded with sewage, and autoclaved Nile water. The results indicate the fecal streptococci were able to persist and multiply in Nile water containing Bladex or Etazine and seeded with sewage. The successive addition of Bladex and Etazine (up to 10 mg/liter) increased its degradation rate and had no inhibitory effect on the bacterial parameters.     

高效油水分离器和过滤器处理油田污水的现场实验研究

为了开发高效、低耗的含油污水处理工艺技术和设备,在吉林油田新立采油厂,对石油大学（北京）研制开发的高效油水分离器和高效过滤器处理油田污水的技术性能进行了现场测试研究。油水分离器采用斜通道波纹板作内构件,过滤器用炉渣做滤料。现场实验表明：高效油水分离器和高效过滤器串联处理油田污水,不论污水含油量高低,其过滤器出口水中含油量不高于１０ｍｇ／Ｌ者达９４％：过滤器出口水中悬浮物不高于１０ｍｇ／Ｌ者占８６％。结果说明该套装置在油和悬浮物的脱除及操作方面性能优异。有推广应用前景。     

AN ANALYSIS OF COLIFORM BACTERIA IN THE UPPER ILLINOIS WATERWAY1

ABSTRACT: During the summer of 1971 about 150 water samples were examined for total and fecal coliform bacteria in the Upper Illinois Waterway at 19 river stations. The data per station were found to be normal geometric distributed. Bacteria densities changed with sampling dates and generally decreased with water movement downstream. Several sewage treatment effluents made marked pulses along the bacterial die-off curves. The observed fecal coliform results were evaluated in terms of the Illinois Pollution Control Board's standards. The FC:TC ratio on the waterway for each station were presented. Using Chick's Law, coliforms death rates were estimated. Efforts to correlate observed total and fecal densities with temperature, flow, algal densities, dissolved oxygen, and 5-day biochemical oxygen demand were not successful. (KEY TERMS: algae; biochemical oxygen demand; coliform bacteria; dissolved oxygen; flow; stream survey; temperature; water pollution; water quality standards)     

GROUNDWATER QUALITY IN THE CORTARO AREA NORTHWEST OF TUCSON,ARIZONA1

ABSTRACT The Cortaro Area is currently the depository for much of the liquid waste from the City of Tucson. In the past, more than one-half of the sewage effluent was used for crop irrigation. However, since 1970 virtually all of the sewage effluent has been percolated in the normally dry Santa Cruz River channel. Nitrate and chloride contents were monitored monthly in water samples from about 20 large-capacity irrigation wells. Contents and seasonal trends for these constituents were closely related to the disposal of sewage effluent. Water quality problems other than nitrate include total dissolved solids, boron, coliform, and lead. High lead contents in the area appear to be a natural phenomenon and the coliform contents are likely related to poor well construction. The other quality problems are primarily due to sewage effluent.     

Sorption-desorption of carbamazepine from irrigated soils

Th anti-seizure medication carbamazepine is often found in treated sewage effluent and environmental samples. Carbamazepine has been shown to be very persistent in sewage treatment, as well as ground water. Due to environmental persistence, irrigation with sewage effluent could result in carbamazepine contamination of surface and ground water. To determine the potential for leaching of carbamazepine, a series of adsorption and desorption batch equilibrium experiments were conducted on irrigated soils. It was found that carbamazepine adsorption to biosolid-amended (T) soils had a KD of 19.8 vs. 12.6 for unamended soil. Based on adsorption, carbamazepine leaching potential would be categorized as low. During desorption significant hysteresis was observed and KD increased for both soils. Desorption isotherms also indicate a potential for irreversibly bound carbamazepine in the T soil. Results indicate that initial removal of carbamazepine via adsorption from irrigation water is significant and that desorption characteristics would further limit the mobility of carbamazepine through the soil profile indicating that carbamazepine found in sewage effluent used for irrigation has a low leaching potential.     

Survival potential of Escherichia coli and Enterococci in subtropical beach sand: implications for water quality managers

Fecal bacteria have traditionally been used as indicator organisms to monitor the quality of recreational waters. Recent work has questioned the robustness of traditional indicators, particularly at seawater bathing beaches. For example, a study of Florida beaches found unexpectedly high abundances of Escherichia coli, fecal coliforms, and enterococci in beach sand. The aim of the present study was to explain these abundances by assessing the survival of E. coli and enterococci in beach sand relative to seawater. We used a combination of quantitative laboratory mesocosm experiments and field observations. Results suggested that E. coli and enterococci exhibited increased survivability and growth in sand relative to seawater. Because fecal bacteria are capable of replicating in sand, at least under controlled laboratory conditions, the results suggest that sand may be an important reservoir of metabolically active fecal organisms. Experiments with "natural" mesocosms (i.e., unsterilized sand or water rich in micropredators and native bacteria) failed to show the same increases in fecal indicators as was found in sterile sand. It is postulated that this was due to predation and competition with indigenous bacteria in these "natural" systems. Nonetheless, high populations of indicators were maintained and recovered from sand over the duration of the experiment as opposed to the die-off noted in water. Indicator bacteria may wash out of sand into shoreline waters during weather and tidal events, thereby decreasing the effectiveness of these indicators as predictors of health risk and complicating the interpretations for water quality managers.     

A long-term study comparing membrane filtration with Colilert defined substrates in detecting fecal coliforms and Escherichia coli in natural waters

Assessment methods for determining the presence and number of fecal bacteria and Escherichia coli (E. coli) in waters, foodstuffs, sewage effluent, and soils have evolved from multiple tube fermentations (MTF's) to membrane filtrations (MF's) to, most recently, defined substrate technologies (DST's). Mounting evidence indicates Colilert DST (IDEXX, Westbrook, ME) to be a versatile assessment technique for detecting and enumerating E. coli over a range of applications. This study compared the performance of Colilert DST with a confirmed standard MF technique using m-FC broth (Millipore, Bedford, MA) in assessing E. coli in ten different environmental water samples obtained monthly over a 3-year period from the upper Appomattox River, VA. For the duration of the study, E. coli counts measured by Colilert DST were positively correlated (Pearson's correlation coefficient=0.956; slope=0.979; p<0.0001) with E. coli counts measured by confirmed MF procedures. The results of a two-factor ANOVA revealed that Colilert DST counts compared equally to confirmed MF counts by year (p=0.974), by stream sampled (p=1.0), and by season (p=0.696). E. coli counts were significantly lower during cold season months (Dec/Jan/Feb) than during warm season months (Jun/Jul/Aug) for each year contributing to marked variation in sample quality. Counts obtained by Colilert DST compared equally to those obtained by MF across all samples and dates for the three years. Colilert DST presents a laboratory protocol that is simpler to manage, quicker to process, and easier to quantify results than MF. These factors, plus the enhanced precision and versatility of Colilert DST over the span of this three-year study attests to its suitability for testing ambient surface waters.     

Bacterial source tracking guides management of boat head waste in a coastal resort area

Fecal contamination of water bodies causes a public health problem and economic loss. To control such contamination management actions need to be guided by sound science. From 2007–2009 a study was undertaken to determine the sources of fecal bacteria contamination to the marine waters adjoining the Town of Wrightsville Beach, North Carolina, USA. The research effort included sampling for fecal coliform and Enterococcus bacteria, sampling for optical brighteners, dye studies, and use of molecular bacterial source tracking techniques including polymerase chain reaction (PCR) and terminal restriction fragment polymorphism (T-RFLP) fingerprinting of the Bacteroides–Prevotella group. Of the 96 samples collected from nine locations during the study, the water contact standard for Enterococcus was exceeded on 13 occasions. The T-RFLP fingerprint analyses demonstrated that the most widespread source of fecal contamination was human, occurring in 38% of the samples, with secondary ruminant and avian sources also detected. Optical brightener concentrations were low, reflecting a lack of sewage line leakage or spills. A lack of sewer leaks and lack of septic systems in the town pointed toward discharge from boat heads into the marine waters as the major cause of fecal contamination; this was supported by dye studies. Based on these data, the Town initiated action to have the U.S. Environmental Protection Agency declare the coastal waters (out to 3 nautical miles), the nearby Atlantic Intracoastal Waterway and its tributaries a no-discharge zone (NDZ) to alleviate the human fecal pollution. The Town garnered supporting resolutions from other local communities who jointly petitioned the North Carolina Department of Environmental and Natural Resources. This State regulatory agency supported the local government resolutions and sent an application for an NDZ to the EPA in April 2009. The EPA concurred, and in February 2010 the coastal waters of New Hanover County, NC, became the first marine area on the U.S. eastern seaboard between Delaware and the Florida Keys to be declared a no-discharge zone.     

Isolation of salmonella bacteriophages from swine effluent lagoons

Bacteriophages (phages) associated with Salmonella were collected from nine swine manure lagoons in Mississippi. Phages were isolated by an enrichment protocol or directly from effluent. For enrichment, chloroform-treated samples were filtered (0.22 mum) and selectively enriched by adding a cocktail of Salmonella strains in trypticase soy broth. After overnight incubation at 35 degrees C, chloroform was added and samples stored at 5 degrees C. Enriched samples were tested by double agar layer (DAL) plaque assay against individual Salmonella isolates. Phage titers of 2.9 x 10(8) to 2.1 x 10(9) plaque forming units (pfu) per mL were produced, but estimation of phage titers in lagoons was not possible. For direct isolation, effluent was clarified by centrifugation, filtered (0.22 microm), and used in DAL plaque assays to select single-plaque isolates for 15 Salmonella strains. Plaque counts varied among Salmonella strains and lagoons. The most sensitive strain for direct phage recovery was ATCC 13311. Phage titers estimated by direct isolation with ATCC 13311 ranged among lagoons from 12 to 148 pfu per mL. In limited host range tests, 66 isolates recovered by the enrichment protocol produced plaques only on Enteritidis and Typhimurium strains of Salmonella and none produced plaques on lagoon isolates of Citrobacter, Escherichia, Proteus, Providencia, or Serratia. Electron microscopy (EM) showed purified enrichment isolates had Podoviridae morphology (tailless 50-nm icosahedral heads with tail spikes). Electron microscopy of clarified concentrated effluent showed 5.5:1 tailless to tailed phages. The isolated phages have potential as typing reagents, specific indicators, and biocontrol agents of Salmonella.