Screening of antimycotics in Swedish sewage treatment plants - Waters and sludge

Concentrations of six pharmaceutical antimycotics were determined in the sewage water, final effluent and sludge of five Swedish sewage treatment plants (STPs) by solid phase extraction, liquid/solid extraction, and liquid chromatography-electrospray tandem mass spectrometry. The antimycotics were quantified by internal standard calibration. The results were used to estimate national flows that were compared to predictions based on sales figures. Fluconazole was the only one of the six investigated antimycotics that was detected (at concentrations ranging from 90 to 140 ng L⁻¹) in both raw sewage water and final effluent. Negligible amounts of this substance were removed from the aqueous phase, and its levels were below the limit of quantification in all of the analyzed sludge samples. In contrast, clotrimazole, ketoconazole and econazole were present in all of the sludge samples, at concentrations ranging between 200 and 1000 μg kg⁻¹, dry weight. There were close correlations between the national measured and predicted antimycotic mass flows. Antimycotic fate analysis, based on sales figures, indicated that 53% of the total amount of fluconazole sold appeared in the final effluents of the STPs, while 1, 155, 35, 209 and 41% of the terbinafine, clotrimazole, ketoconazole, econazole and miconazole sold appeared in the digested dewatered sludge.     

Antimicrobial resistance in Enterococcus spp. isolated in inflow, effluent and sludge from municipal sewage water treatment plants

Antimicrobial resistance of enterococci was investigated in 42 samples of crude inflow, treated effluent and sludge collected in 14 municipal sewage treatment plants of Portugal. A total of 983 enterococci were recovered and tested, using the diffusion agar method, regarding their sensitivity to 10 different antimicrobial drugs. Multidrug resistance was present in 49.4% of the isolates. Only 3.3% and 0.6% of the investigated strains were resistant to ampicillin and vancomycin, respectively. Resistances found against rifampicin (51.5%), tetracycline (34.6%), erythromycin (24.8%) and nitrofurantoin (22.5%), are causes for substantial concern. Almost 14% of isolates were resistant to ciprofloxacin. Wastewater treatment resulted in enterococci decrease between 0.5 and 4log; nevertheless, more than 4.4 x 10(5)CFU/100ml were present in the outflow of the plants. Our data indicate that the use of antimicrobials had created a large pool of resistance genes and that sewage treatment processes are unable to avoid the dissemination of resistant enterococci into the environment.     

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.     

Natural freezing as a wastewater treatment method: E. coli inactivation capacity

Inactivation capacity of E. coli (strain ATCC 15597) in water by natural freezing was examined via two freezing methods: spray freezing and freezing in a freezer. The effect of freezing temperature (-5, -15 and -35 degrees C), storage time, freeze-thaw cycles on the survival of the test organism were investigated. In addition, the number of cells injured by the freezing process was also examined by using different growth media. The bacteria frozen at the warmer temperature (-5 degrees C) was most sensitive to storage and freeze-thaw cycles as compared to those frozen at -15 and -35 degrees C. In general, greater inactivation efficiencies were achieved under longer storage time and warmer freezing temperature conditions. Freezing and thawing caused cell injury. More cells were injured when frozen at -15 degrees C. The percentage of cells injured decreased as freeze-thaw cycles increased. The spray-freezing process was found more effective in killing the cells. On average, the log reduction rate for the spray ice with two-day storage time was about 4 log units higher than those without any storage after freezing. The results indicated that the natural freezing processes are not only cost-effective techniques for chemical and physical contaminant removal from wastewater or enhancing sludge dewaterability in cold regions but also effective in reducing E. coli concentration.     

Probability of detecting and quantifying faecal contaminations of drinking water by periodically sampling for E. coli: a simulation model study

Drinking water supply companies monitor the presence of Escherichia coli in drinking water to verify the effectiveness of measures that prevent faecal contamination of drinking water. Data are lacking, however, on the sensitivity of the monitoring programmes, as designed under the EU Drinking Water Directive. In this study, the sensitivity of such a monitoring programme was evaluated by hydraulic model simulations of contamination events and calculations of the detection probability of the actual sampling programme of 2002. In the hydraulic model simulations of 16-h periods of 1l h(-1) ingress of untreated domestic sewage, the spread of the contamination through the network and the E. coli concentration dynamics were calculated. The results show that when large parts of the sewage reach reservoirs, e.g. when they originate from the treatment plant or a trunk main, mean detection probabilities are 55-65%. When the contamination does not reach any of the reservoirs, however, the detection probability varies from 0% (when no sampling site is reached) to 13% (when multiple sites are reached). Mean detection probabilities of nine simulated ingress incidents in mains are 5.5% with an SD of 6.5%. In reality, these detection probabilities are probably lower as the study assumed no inactivation or clustering of E. coli, 100% recovery efficiency of the E. coli detection methods and immediate mixing of contaminations in mains and reservoirs. The described method provides a starting point for automated evaluations and optimisations of sampling programmes.     

UV inactivation and characteristics after photoreactivation of Escherichia coli with plasmid: health safety concern about UV disinfection

Occurrence and degree of photoreactivation after ultraviolet (UV) exposure have been widely studied. However, the characteristics of photoreactivated microorganisms were rarely investigated. Hence, in this study, Escherichia coli with plasmids of ampicillin (amp)-resistance or fluorescence was used as indicators to examine the UV inactivation efficiencies and variations of characteristics of E. coli after subsequent photoreactivation.The experimental results indicate that the amp-resistant bacteria and the fluorescent bacteria used in this study had similar trends of UV dose-response curves. 3.5-log₁₀ and 3-log₁₀ reductions were achieved with a UV dose of 5 mJ/cm² for the amp-resistant and fluorescent E. coli, respectively. There was no significant difference in the UV inactivation behavior, as compared with common strains of E. coli.For the amp-resistant E. coli and the fluorescent E. coli, after exposures with UV doses of 5, 15, 25, 40 and 80 mJ/cm², the corresponding percent photoreactivations after a 4 h exposure to photoreactivating light were 1% and 46% respectively for a UV dose of 5 mJ/cm², and essentially negligible for all other UV doses. Furthermore, the photoreactivated amp-resistant bacteria still have the ability of amp-resistance. And the revived fluorescent E. coli showed similar fluorescent behavior, compared with the untreated bacteria. The experimental results imply that after UV inactivation and subsequent photoreactivation, the bacteria retained the initial characteristics coded in the plasmid. This reveals a possibility that some characteristics of bacteria can retain or recover through photoreactivation, and a safety concern about pathogenicity revival might need to be considered with UV disinfection and photoreactivation.     

Linear correlation between inactivation of E. coli and OH radical concentration in TiO2 photocatalytic disinfection

The biocidal action of the TiO2 photocatalyst has been now well recognized from massive experimental evidences, which demonstrates that the photocatalytic disinfection process could be technically feasible. However, the understanding on the photochemical mechanism of the biocidal action largely remains unclear. In particular, the identity of main acting photooxidants and their roles in the mechanism of killing microorganisms is under active investigation. It is generally accepted that reactive oxygen species (ROS) and OH radicals play the role. The aim of this study is to determine how the OH radical, acting either independently or in collaboration with other ROS, is quantitatively related to the inactivation of E. coli. The steady-state concentrations of OH radicals ([*OH]ss) in UV-illuminated TiO2 suspensions could be quantified from the measured photocatalytic degradation rates of p-chlorobenzoic acid (a probe compound) and its literature bimolecular rate constant with OH radicals. The results demonstrated an excellent linear correlation between [*OH]ss and the rates of E. coli inactivation, which indicates that the OH radical is the primary oxidant species responsible for inactivating E. coli in the UV/TiO2 process. The CT value of OH radical for achieving 2 log E. coli inactivation was initially found to be 0.8x10(-5) mg min/l, as predicted by the delayed Chick-Watson model. Although the primary role of OH radicals in photocatalytic disinfection processes has been frequently assumed, this is the first quantitative demonstration that the concentration of OH radicals and the biocidal activity is linearly correlated.     

Bacterial pathogen incidences in sludge from Swedish sewage treatment plants

This study surveyed the presence of bacterial pathogens in eight Swedish sewage treatment plants (STPs), with four different treatment methods, focusing on detection of zoonotic bacteria in raw and treated sludge. Salmonella spp., Listeria monocytogenes, Campylobacter coli and jejuni, Escherichia coli O157 and indicator bacteria were investigated. Samplings were performed from July 2000 to June 2002, resulting in 64 raw sludge samples and 69 treated sludge samples. The samples from raw sludge (67%) and treated sludge (55%) were positive for Salmonella; 49 different serotypes were detected. Restriction enzyme analysis and pulsed field gel electrophoresis of Salmonella serotypes indicated that Salmonella persists in STPs and that there is a continuous supply of new strains. There are differences in treatment methods concerning the reduction of pathogens and indicator bacteria. If spread on arable land, sludge increases the environmental load of pathogens; this increases the risk for spreading diseases to people and animals.     

Determination of sorption of seventy-five pharmaceuticals in sewage sludge

Sorption of 75 active pharmaceutical ingredients (APIs) to three different types of sludge (primary sludge, secondary sludge with short and long sludge age respectively) were investigated. To obtain the sorption isotherms batch studies with the APIs mixture were performed in four nominal concentrations to water containing 1 g of sludge. The range of APIs concentrations was between ng L⁻¹ to μg L⁻¹ which are found in the wastewater effluents. Isotherms were obtained for approximately 45 of the APIs, providing distribution coefficients for linear (K_d), Freundlich (K_f) and Langmuir (K_L) isotherms. K_d, K_f and K_L ranging between 7.1 × 10⁴ and 3.8 × 10⁷, 1.1 × 10⁻² and 6.1 × 10⁴ and 9.2 × 10⁻³ and 1.1 L kg⁻¹, respectively. The obtained coefficients were applied to estimate the fraction of APIs in the water phase (see Abstract Graphic). For 37 of the 75 APIs, the predicted presence in the liquid phase was estimated to >80%. 24 APIs were estimated to be present in the liquid phase between 20 and 80%, and 14 APIs were found to have <20% presence in the liquid phase, i.e. high affinity towards sludge. Furthermore, the effect of pH at values 6, 7 and 8 was evaluated using one way ANOVA-test. A significant difference in K_ds due to pH changes were found for 6 of the APIs (variation 10-20%).     

Fate of natural estrogens in batch mixing experiments using municipal sewage and activated sludge

Since natural estrogens such as 17beta-estradiol (E2) and estrone (E1) are excreted daily by humans, E2 and E1, which are classified as inevitable endocrine-disrupting chemicals, are always present in sewage wastewater. For several years, the monitoring and removal of natural estrogens at sewage treatment plants have been examined by many investigators. However, little is known regarding the exact behavior of estrogens in actual sewage when in contact with activated sludge. In this study, the fate of E2 and E1 as a result of adsorption and decomposition in batch mixing experiments using municipal wastewater and activated sludge collected from an actual municipal sewage treatment plant was investigated. Estrogen concentrations were determined using an enzyme-linked immunosorbent assay (ELISA) kit. E2 and E1 in sewage were removed from the liquid phase in contact with activated sludge, and E2 and E1 adsorbed on the sludge were decomposed in 4h. Significant changes in the adsorption and decomposition of E2 and E1 on the sludge were not observed at low temperatures or when different sludge samples were used such as those acclimatized to low-loading and high-loading conditions. In contrast, the processes leading to the removal of estrogens, such as the adsorption and decomposition of estrogens in contact with activated sludge, were inactivated by sterilizing the sludge. Natural estrogens adsorb onto the activated sludge, and are therefore easy to be biodegraded. In a biological reaction process, therefore, the estrogens will be rapidly removed at the initial stage, when the sewage is satisfactorily mixed with the sludge.     

Confirmation of E. coli among other thermotolerant coliform bacteria in paper mill effluents, wood chips screening rejects and paper sludges

Paper sludges are solid wastes material generated from the paper production, which have been characterized for their chemical contents. Some are rich in wood fiber and are a good carbon source, for example the primary and de-inking paper sludges. Others are made rich in nitrogen and phosphorus by pressing the activated sludge, resulting from the biological water treatments, with the primary sludge, yielding the combined paper sludge. Still, in the absence of sanitary effluents very few studies have addressed the characterization of their coliform microflora. Therefore, this study investigated the thermotolerant coliform population of one paper mill effluent and two paper mill sludges and wood chips screening rejects using chromogenic media. For the first series of analyses, the medium used was Colilert broth and positive tubes were selected to isolate bacteria in pure culture on MacConkey agar. In a second series of analyses, double selective media, based on ss-galactosidase and ss-glucuronidase activities, were used to isolate bacteria. First, the presence of thermotolerant coliforms was detected in low numbers in most water effluents, but showed that the entrance of the thermotolerant coliforms was early in the industrial process. Also, large numbers of thermotolerant coliforms, i.e., 7,000,000 MPN/g sludge (dry weight; d.w.), were found in combined sludges. From this first series of isolations, bacteria were purified on MacConkey medium and identified as Citrobacter freundii, Enterobacter sp, E. sakazakii, E. cloacae, Escherichia coli, K. pneumoniae, K. pneumoniae subsp. rhinoscleromatis, K. pneumoniae subsp. ozaenae, K. pneumoniae subsp. pneumoniae, Pantoea sp, Raoultella terrigena, R. planticola. Second, the presence of thermotolerant coliforms was measured at more than 3,700-6,000 MPN/g (d.w) sludge, whereas E. coli was detected from 730 to more than 3,300 MPN/g (d.w.) sludge. The presence of thermotolerant coliform bacteria and E. coli was sometimes detected from wood chips screening rejects in large quantities. Also, indigenous E. coli were able to multiply into the combined sludge, and inoculated E. coli isolates were often able to multiply in wood chips and combined sludge media. In this second series of isolations, API20E and Biolog identified most isolates as E. coli, but others remained unidentified. The sequences of the 16S rDNA confirmed that most isolates were likely E. coli, few Burkholderia spp, but 10% of the isolates remained unidentified. This study points out that the coliform bacteria are introduced by the wood chips in the water effluents, where they can survive throught the primary clarifier and regrow in combined sludges.     

城市污水厂污泥处置技术分析及福州市污泥处置的思考

本文对城市污水厂污泥处理处置主要技术进行分析,并阐述了国外污泥处置技术发展趋势,提出了污泥处理处置工作急需解决的问题。结合当前的污泥处置技术发展趋势,通过对福州市污水厂污泥处置现状及存在问题分析,提出了福州市今后的污泥处置技术路线:将厌氧消化做为有效减容、减量的手段,对污染物指标达到国家土地利用标准的污泥考虑土地利用,污染物指标不符合国家土地利用标准的污泥考虑焚烧和建材利用。     

Seasonal change and fate of coliphages infected to Escherichia coli O157:H7 in a wastewater treatment plant

Seasonal change of virulent phage infected to two E. coli O157:H7 strains (O:157-phage) in the influent of a domestic wastewater treatment plant in the central part of Japan and fate of O:157-phage in the plant were monitored almost monthly from March 2001 to February 2002. Coliphage infected to nonpathogenic E. coli O157:H7 ATCC43888 (43888-phage) was detected for 1 year. On the other hand, phage infected to pathogenic E. coli O157:H7 EDL933 (EDL-phage) was detected intermittently. Concentration of EDL-phage was almost one-tenth of that of 43888-phage. The progressive decrease in phage concentration with the treatment steps was observed. No phage was detected in the supernatant from the secondary settling tank and effluent. PCR amplification of the Stx 2 gene that encodes Shiga toxin (Stx) was observed when O:157-phage concentration in the influent was high x10(3) PFU/ml order. Concentration and percentage of suspended O:157-phage decreased with the progress of the wastewater treatment. 933W phage, which encodes Stx 2 gene, was more fragile and sensitive to chlorination than T4 phage. However, addition of 0.02 mg/l chlorine, in conformance with the required concentration of the plant, did not affect the viability of T4 and 933 W phages. On the other hand, 1mg/l chlorine inactivated the 933 W phage significantly.     

ESBL-producing E. coli in Austrian sewage sludge

The aim of this study was to investigate the degree of contamination of sewage sludge with ESBL-producing Escherichia coli strains and the effectiveness of different sewage sludge treatment methods.Monthly sewage sludge samples were collected between January and September 2009 in 5 different sewage treatment plants and tested for the presence of ESBL E. coli. In addition, the number of colony forming units (CFU) of E. coli and coliform bacteria before and after the different sludge treatment methods (aerobic/anaerobic digestion, lime stabilization, and thermal treatment) was investigated.Of the 72 sewage sludge samples investigated, ESBL-positive E. coli were found in 44 (61.1%) sewage sludge samples. The classification of β-lactamase groups was carried out in 15 strains resulting in the detection of 2 different groups (CTX-M and TEM) of bla genes. All 15 of them had a CTX-M gene and 4 of these strains furthermore carried a TEM gene.With regard to the CFU of E. coli and coliform bacteria, thermal treatment and lime stabilization following dehydration sufficiently reduced pathogen concentrations. The plants using merely stabilization and dehydration showed an increase of E. coli and coliform bacteria and thus also an increase in ESBL-producing E. coli.     

Effect of Antarctic solar radiation on sewage bacteria viability

The majority of coastal Antarctic research stations discard untreated sewage waste into the near-shore marine environment. However, Antarctic solar conditions are unique, with ozone depletion increasing the proportion of potentially damaging ultraviolet-B (UV-B) radiation reaching the marine environment. This study assessed the influence of Antarctic solar radiation on the viability of Escherichia coli and sewage microorganisms at Rothera Research Station, Adelaide Island, Antarctic Peninsula. Cell viability decreased with increased exposure time and with exposure to shorter wavelengths of solar radiation. Cell survival also declined with decreasing cloud cover, solar zenith angle and ozone column depth. However, particulates in sewage increased the persistence of viable bacteria. Ultraviolet radiation doses over Rothera Point were highest during the austral summer. During this time, solar radiation may act to partially reduce the number of viable sewage-derived microorganisms in the surface seawater around Antarctic outfalls. Nevertheless, this effect is not reliable and every effort should be made to fully treat sewage before release into the Antarctic marine environment.     

Survival of Escherichia coli in two sewage treatment plants using UV irradiation and chlorination for disinfection

We investigated the survival of Escherichia coli in two STPs utilising UV irradiation (STP-A) or chlorination (STP-B) for disinfection. In all, 370 E. coli strains isolated from raw influent sewage (IS), secondary treated effluent (STE) and effluent after the disinfection processes of both STPs were typed using a high resolution biochemical fingerprinting method and were grouped into common (C-) and single (S-) biochemical phenotypes (BPTs). In STP-A, 83 BPTs comprising 123 isolates were found in IS and STE, of which 7 BPTs survived UV irradiation. Isolates tested from the same sites of STP-B (n = 220) comprised 122 BPTs, however, only two BPTs were found post-chlorination. A representative isolate from each BPT from both STPs was tested for the presence of 11 virulence genes (VGs) associated with uropathogenic (UPEC) or intestinal pathogenic (IPEC) E. coli strains. Strains surviving UV irradiation were distributed among seven phylogenetic groups with five BPTs carrying VGs associated with either UPEC (4 BPTs) or IPEC (1 BPT). In contrast, E. coli strains found in STP-B carried no VGs. Strains from both STPs were resistant to up to 12 out of the 21 antibiotics tested but there was no significant difference between the numbers of antibiotics to which surviving strains were resistant to in these STPs. Our data suggests that some E. coli strains have a better ability to survive STPs utilising chlorination and UV irradiation for disinfection. However, strains that survive UV irradiation are more diverse and may carry more VGs than those surviving SPTs using chlorination.     

The effects of temperature, pH, and ammonia concentration on the inactivation of Ascaris eggs in sewage sludge

The reported inactivation of Ascaris eggs during alkaline sludge stabilization is highly variable. The objective of our research was to better understand the sources of this variability by quantifying the effects of temperature, pH, and ammonia concentration on the inactivation of indigenous Ascaris eggs in wastewater sludge. Primary sludge was supplemented with ammonia (0, 1000, and 5000mg/l NH(3)-N) and Ca(OH)(2) and incubated in sealed bottles across the range of temperatures (20, 30, 40, and 50 degrees C) and pH (7 and 12) that may be encountered during treatment. Changes in egg viability over time were fit to a two-parameter kinetic model (shoulder and first-order region); to compare treatment conditions, the time for 99% inactivation (t(99)) was also calculated. Each 10 degrees C increase in temperature caused a significant decrease in t(99) at every pH and ammonia concentration tested. At 50 degrees C, the effect of temperature was dominant, such that no effect of pH or ammonia was observed. At 30 and 40 degrees C, raising the pH from 7 to 12 decreased t(99), but at 20 degrees C no pH effect was seen over 80 d (very little inactivation occurred). At 20, 30, and 40 degrees C, the addition of ammonia dramatically decreased t(99). The effect of pH could not be completely separated from that of ammonia, as the unamended sludge samples contained 100-200mg/l indigenous ammonia. Because temperature, pH, and ammonia all contributed to Ascaris egg inactivation, it is essential that these parameters are measured and accounted for when assessing the effectiveness of alkaline stabilization. Furthermore, inactivation by ammonia could be exploited to improve the effectiveness of alkaline sludge stabilization.     

Population structure of Cladophora-borne Escherichia coli in nearshore water of Lake Michigan

We previously reported that the macrophytic green alga Cladophora harbors high densities (up to 10(6) colony-forming units/g dry weight) of the fecal indicator bacteria, Escherichia coli and enterococci, in shoreline waters of Lake Michigan. However, the population structure and genetic relatedness of Cladophora-borne indicator bacteria remain poorly understood. In this study, 835 E. coli isolates were collected from Cladophora tufts (mats) growing on rocks from a breakwater located within the Indiana Dunes National Lakeshore in northwest Indiana. The horizontal fluorophore enhanced rep-PCR (HFERP) DNA fingerprinting technique was used to determine the genetic relatedness of the isolates to each other and to those in a library of E. coli DNA fingerprints. While the E. coli isolates from Cladophora showed a high degree of genetic relatedness (92% similarity), in most cases, however, the isolates were genetically distinct. The Shannon diversity index for the population was very high (5.39). Both spatial and temporal influences contributed to the genetic diversity. There was a strong association of isolate genotypes by location (79% and 80% for lake- and ditch-side samplings, respectively), and isolates collected from 2002 were distinctly different from those obtained in 2003. Cladophora-borne E. coli isolates represented a unique group, which was distinct from other E. coli isolates in the DNA fingerprint library tested. Taken together, these results indicate that E. coli strains associated with Cladophora may be a recurring source of indicator bacteria to the nearshore beach.     

Escherichia coli, enterococci, and Bacteroides thetaiotaomicron qPCR signals through wastewater and septage treatment

Fecal indicators such as Escherichia coli and enterococci are used as regulatory tools to monitor water with 24 h cultivation techniques for possible input of sewage or feces and presence of potential enteric pathogens yet their source (human or animal) cannot be determined with routine methods. This critical uncertainty has furthered water pollution science toward new molecular approaches. Members of Bacteroides genus, such as Bacteroides thetaiotaomicron are found to have features that allow their use as alternative fecal indicators and for Microbial Source Tracking (MST). The overall aim of this study was to evaluate the concentration and fate of B. thetaiotaomicron, throughout a wastewater treatment facility and septage treatment facility. A large number of samples were collected and tested for E. coli and enterococci by both cultivation and qPCR assays. B. thetaiotaomicron qPCR equivalent cells (mean: 1.8 × 10⁷/100 mL) were present in significantly higher concentrations than E. coli or enterococci in raw sewage and at the same levels in raw septage. The removal of B. thetaiotaomicron target qPCR signals was similar to E. coli and enterococci DNA during the treatment of these wastes and ranged from 3 to 5 log₁₀ for wastewater and was 7 log₁₀ for the septage. A significant correlation was found between B. thetaiotaomicron marker and each of the conventional indicators throughout the waste treatment process for both raw sewage and septage. A greater variability was found with enterococci when compared to E. coli, and CFU and equivalent cells could be contrasted by various treatment processes to examine removal and inactivation via septage and wastewater treatment. These results are compared and contrasted with other qPCR studies and other targets in wastewater samples providing a view of DNA targets in such environments.     

Accumulation and fate of green fluorescent labeled Escherichia coli in laboratory-scale drinking water biofilters

Biological filters combining microbial activity and rapid sand filtration are used in drinking water treatment plants for enhanced biodegradable organic matters (BOM) removal. Biofilms formed on filter media comprised of bacteria enclosed in a polymeric matrix are responsible for the adsorption of BOM and attachment of planktonic microorganisms. This study investigated the removal of Escherichia coli cells injected into laboratory-scale biofilters and the role of biofilm in retaining the injected E. coli. Green fluorescent protein was used as a specific marker to detect and quantify E. coli in the biofilms. About 35% of the total injected E. coli cells were observed in the filter effluents, when initial cell concentrations were measured at 7.4 x 10(6) CFU/mL and 1.6 x 10(7) CFU/mL in two separate experiments. The results from real-time PCR and plate count analysis indicated that 95% of the E. coli retained inside the filters were either non-viable or could not be recovered by colony counting techniques. Injected cells were unevenly distributed inside the filter with more than 70% located at the top 1/5 of the filter. Images obtained from an epifluorescent microscope showed that E. coli cells were embedded inside the biofilm matrix and presented mainly as microcolonies intertwined with other microorganisms, which was consistent with findings from standard plate count methods and qPCR.