Assessment of human adenovirus removal in a full-scale membrane bioreactor treating municipal wastewater

Human adenoviruses (HAdVs) in wastewater samples taken from four different treatment stages of a full-scale municipal wastewater treatment plant (i.e., incoming raw sewage, primary sedimentation effluent, membrane bioreactor (MBR) influent, and MBR effluent) were quantified by real-time PCR assays to further estimate removal efficiency of the HAdVs. Based on hexon gene sequence comparisons, HAdV species A, C, and F were consistently found in the wastewater samples. In general, all three identified HAdV species were detected in most of the wastewater samples using the real-time PCR assays. Overall HAdV concentrations were rather stable over the entire 8-month study period (January-August, 2008) (approximately 10⁶-10⁷ viral particles/L of wastewater for the raw sewage and primary effluent; 10⁸-10⁹ viral particles/L for the MBR influent; and, 10³-10⁴ viral particles/L for the MBR effluent). No significant seasonal differences were noticed for the HAdV abundances. Removal efficiencies of the viral particles in the full-scale MBR process were assessed and showed an average HAdV removal of 5.0 ± 0.6 logs over the study period. The removal efficiencies for F species (average log removal of 6.5 ± 1.3 logs) were typically higher (p-value <0.05) than those of the other two species (average of 4.1 ± 0.9 and 4.6 ± 0.5 logs for species A and C, respectively). These results demonstrate that the full-scale MBR system efficiently removed most HAdV from the wastewater leaving about 10³ viral particles/L in the MBR effluent.     

Quantification and molecular characterization of enteric viruses detected in effluents from two hospital wastewater treatment plants

Hospital wastewater has been described as an important source of spreading pathogenic microorganisms in the environment. However, there are few studies reporting the presence and concentrations of gastroenteric viruses and hepatitis A viruses in these environmental matrices. The aim of this study was to assess the contamination by viruses responsible for acute gastroenteritis and hepatitis derived from hospital wastewater treatment plants (WWTPs). Rotavirus A (RV-A), human adenoviruses (HAdV), norovirus genogroup I and II (NoV GI/GII) and hepatitis A viruses (HAV) were detected and quantified in sewage samples from two WWTPs located in Rio de Janeiro (Brazil) that operates different sewage treatments. WWTP-1 uses an Upflow Anaerobic Sludge Blanket (UASB reactor) and three serial anaerobic filters while WWTP-2 uses aerobic processes, activated sludge with extended aeration and final chlorination of the effluents. Viruses’ detection was investigated by using conventional PCR/RT-PCR, quantitative real-time PCR (qPCR) and partial sequencing of the genome of the viruses detected. Rate of viruses detection ranged from 7% (NoV GI in WWTP-1) to 95% (RV-A in WWTP-2) and genome from all viruses were detected. The most prevalent genotypes were RV-A SG I, HAdV species D and F, NoV GII/4 and HAV subgenotype IA. Mean values of viral loads (genome copies (GC)/ml) obtained in filtered effluents from anaerobic process was 1.9 × 10³ (RV-A), 2.8 × 10³ (HAdV) and 2.4 × 10³ (NoV GII). For chlorinated effluents from activated sludge process, the mean values of viral loads (GC/ml) was 1.2 × 10⁵ (RV-A), 1.4 × 10³ (HAdV), 8.1 × 10² (NoV GII) and 2.8 × 10⁴ (HAV). Data on viral detection in treated effluents of hospital WWTPs confirmed the potential for environmental contamination by viruses and could be useful to establish standards for policies on wastewater management.     

Real-time PCR detection of adenoviruses, polyomaviruses, and torque teno viruses in river water in Japan

The prevalence of DNA viruses in water from the Tamagawa River, Japan was quantitatively surveyed for 6 months, from April to September 2003. A total of 18 river water samples were subjected to virus concentration method using an electronegative membrane, followed by DNA extraction and direct quantitative real-time polymerase chain reaction (qPCR) for DNA viruses. Adenoviruses of serotypes 40 and 41 were detected most frequently in the river water samples tested (61.1%), at a concentration ranging from 3.16 × 10³ to 1.38 × 10⁵ copies/l, followed by JC polyomaviruses (11.1%) and torque teno viruses (5.6%). No sample was positive for BK polyomaviruses. In addition, for selective detection of virus particles, adenoviruses 40 and 41 were tested with qPCR combined with an immunomagnetic separation technique; they were detected in only 16.7% of the samples, showing a concentration ranging from 7.42 × 10² to 4.24 × 10⁴ copies/l. This study is significant since it is the first study to demonstrate the prevalence of polyomaviruses in water samples in Japan and to use immunomagnetic separation qPCR to detect adenovirus particles in aquatic environments.     

One year environmental surveillance of rotavirus specie A (RVA) genotypes in circulation after the introduction of the Rotarix® vaccine in Rio de Janeiro, Brazil

Rotavirus specie A (RVA) infection is the leading cause of severe acute diarrhea among young children worldwide. To reduce this major RVA health impact, the Rotarix^® vaccine (GlaxoSmithKline, Rixensart, Belgium) was introduced in the Brazilian Expanded Immunization Program in March 2006 and became available to the entire birth cohort. The aim of this study was to evaluate the spread of RVA in the environment after the introduction of Rotarix^® in Brazil. For this purpose, a Wastewater Treatment Plant (WTP) in Rio de Janeiro was monitored for one year to detect, characterize and discriminate RVA genotypes and identify possible circulation of vaccine strains. Using TaqMan^® quantitative PCR (qPCR), RVA was detected in 100% (mean viral loads from 2.40 × 10⁵ to 1.16 × 10⁷ genome copies (GC)/L) of sewage influent samples and 71% (mean viral loads from 1.35 × 10³ to 1.64 × 10⁵ GC/L) of sewage effluent samples. The most prevalent RVA genotypes were P[4], P[6] and G2, based on VP4 and VP7 classification. Direct nucleotide sequencing (NSP4 fragment) and restriction enzyme digestion (NSP3) analysis did not detect RVA vaccine-like strains from the sewage samples. These data on RVA detection, quantification and molecular characterization highlight the importance of environmental monitoring as a tool to study RVA epidemiology in the surrounding human population and may be useful on ongoing vaccine monitoring programs, since sewage may be a good screening option for a rapid and economical overview of the circulating genotypes.     

Occurrence of enteric viruses and faecal indicators in submarine groundwater discharges in the coastal environment of the Mexican Caribbean

The occurrence of faecal microorganisms such as total coliforms (TCs), Escherichia coli (E. coli), coliphages (somatic and F+ specific) and the genomes of faecal viruses: human adenovirus (HAdV), norovirus (NoV) genogroup I (GI) and II (GII), enterovirus (EV), aichi virus (AiV) and Pepper mild mottle virus (PMMoV), was explored in submarine groundwater discharges (SGDs) in the Mexican Caribbean. Ultra-filtrated water samples were subjected to nucleic acid extraction and real-time quantitative polymerase chain reaction (PCR). Results showed the presence of TC bacteria at all sites, whereas the detection of E. coli occurred only during the dry season. Coliphages occurred during the rainy and dry seasons, with concentrations ranging from 10 to 2560 plaque forming units (PFU/100 ml). HAdV and AiV were detected in 50% and 41% of the samples, at concentrations ranging from 10² to 10⁴ genome copies per litre (GC/L) and from 10² to 10⁶ GC/L, respectively. NoV GI and II were detected in 25% and 16% of the samples, at concentrations of 10¹ GC/L and 10²–10³ GC/L, respectively. PMMoV was detected in 50% of the samples at concentrations ranging from 10¹ to 10³ GC/L. The EV genomes were not detected. These findings demonstrate that faecal microorganisms can be transported through SGDs in the Mexican Caribbean, potentially contributing to human health risks for recreation.     

Linking non-culturable (qPCR) and culturable enterococci densities with hydrometeorological conditions

Quantitative polymerase chain reaction (qPCR) measurement of enterococci has been proposed as a rapid technique for assessment of beach water quality, but the response of qPCR results to environmental conditions has not been fully explored. Culture-based E. coli and enterococci have been used in empirical predictive models to characterize their responses to environmental conditions and to increase monitoring frequency and efficiency. This approach has been attempted with qPCR results only in few studies. During the summer of 2006, water samples were collected from two southern Lake Michigan beaches and the nearby river outfall (Burns Ditch) and were analyzed for enterococci by culture-based and non-culture-based (i.e., qPCR) methods, as well as culture-based E. coli. Culturable enterococci densities (log CFU/100 ml) for the beaches were significantly correlated with enterococci qPCR cell equivalents (CE) (R = 0.650, P < 0.0001, N = 32). Enterococci CE and CFU densities were highest in Burns Ditch relative to the beach sites; however, only CFUs were significantly higher (P < 0.0001). Culturable enterococci densities at Burns Ditch and the beaches were significantly correlated (R = 0.565, P < 0.0001, N = 32). Culturable E. coli and enterococci densities were significantly correlated (R = 0.682, P < 0.0001, N = 32). Regression analyses suggested that enterococci CFU could be predicted by lake turbidity, Burns Ditch discharge, and wind direction (adjusted R² = 0.608); enterococci CE was best predicted by Burns Ditch discharge and log-transformed lake turbidity × wave height (adjusted R² = 0.40). In summary, our results show that analytically, the qPCR method compares well to the non-culture-based method for measuring enterococci densities in beach water and that both these approaches can be predicted by hydrometeorological conditions. Selected predictors and model results highlight the differences between the environmental responses of the two method endpoints and the potentially high variance in qPCR results.     

Enhanced nitrogen and phosphorus removal from eutrophic lake water by Ipomoea aquatica with low-energy ion implantation

Ipomoea aquatica with low-energy N⁺ ion implantation was used for the removal of both nitrogen and phosphorus from the eutrophic Chaohu Lake, China. The biomass growth, nitrate reductase and peroxidase activities of the implanted I. aquatica were found to be higher than those of I. aquatica without ion implantation. Higher NO₃-N and PO₄-P removal efficiencies were obtained for the I. aquatica irradiation at 25 keV, 3.9 × 10¹⁶ N⁺ ions/cm² and 20 keV 5.2 × 10¹⁶ N⁺ ions/cm², respectively (p < 0.05). Moreover, the nitrogen and phosphorus contents in the plant biomass with ion implantation were also greater than those of the controls. I. aquatica with ion implantation was directly responsible for 51-68% N removal and 54-71% P removal in the three experiments. The results further confirm that the ion implantation could enhance the growth potential of I. aquatica in real eutrophic water and increase its nutrient removal efficiency. Thus, the low-energy ion implantation for aquatic plants could be considered as an approach for in situ phytoremediation and bioremediation of eutrophic waters.     

Accumulation of oxytetracycline and norfloxacin from saline soil by soybeans

Soil of former shrimp aquaculture facilities in Thailand may be contaminated by antibiotics (e.g. oxytetracycline and norfloxacin) and have elevated salinity. Therefore, reuse of this land can be problematic. The utility of soybean (Glycine max (L.) Merr.) for phytoremediation was investigated. The rate of germination and seedling emergence in prepared contaminated soil (conductivity 17.7 dS m^− 1 from adding 70 mg sodium chloride g^− 1 dry weight, 105 mg kg^− 1 dry weight oxytetracycline and 55 mg kg^− 1 dry weight norfloxacin) in sunlight was approximately 80% that of uncontaminated soil. This reduction was largely due to the high salinity. The antibiotics of interest degraded relatively rapidly in soil (half-life < 10 h for both) but loss was slower in deionised water. Accumulation of the antibiotics from deionised water by soybean resulted in little effect on growth rate and maximum levels in plants were observed after two days exposure, followed by declining concentrations. For soybean plants grown in saline soil, 90% removal of NaCl from soil adjacent to plant roots was observed, most within two days. Wilting and defoliation occurred, but plants recovered after 10 days and maximum salt levels in plants exceeded 20,000 mg g^− 1 dry weight with translocation from root to shoot tissue noted. Soybean plants also accumulated the antibiotics from prepared contaminated saline soil, but translocation from the roots was not observed. The results showed that soybean can be valuable for phytoremediation in these situations.     

Environmental occurrence, fate and transformation of benzodiazepines in water treatment

Benzodiazepine derivatives are prescribed in large quantities globally and are potentially new emerging environmental contaminants. Unfortunately, a dearth of data exists concerning occurrence, persistence and fate in the environment. This paper redresses this by reviewing existing literature, assessing the occurrence of selected benzodiazepine anxiolytics (diazepam, oxazepam and bromazepam) in wastewater influent and effluent and surface water from Slovenia, evaluating their removal during water treatment and identifying the transformation products formed during water treatment. Their occurrence was monitored in hospital effluent, river water and in wastewater treatment plant influent and effluent. The study reveals the presence of benzodiazepine derivatives in all samples with the highest amounts in hospital effluents: 111 ng L⁻¹, 158 ng L⁻¹ and 72 ng L⁻¹ for diazepam, bromazepam and oxazepam, respectively. Removal efficiencies with respect to biological treatment of diazepam were 16-18% (oxic), 18-32% (anoxic → oxic), 53-76% (oxic → anoxic) and 83% (oxic → anoxic → oxic → anoxic cascade bioreactors), while the removal oxazepam was 20-24% under anoxic conditions. Coupled biological and photochemical treatment followed by the adsorption to activated carbon resulted in a removal efficiency of 99.99%. Results reveal the recalcitrant nature of benzodiazepine derivatives and suggest that only combinational treatment is sufficient to remove them. In addition, eight novel diazepam and four novel oxazepam transformation products are reported.     

Removal of diatrizoate with catalytically active membranes incorporating microbially produced palladium nanoparticles

There is an increasing concern about the fate of iodinated contrast media (ICM) in the environment. Limited removal efficiencies of currently applied techniques such as advanced oxidation processes require more performant strategies. The aim of this study was to establish an innovative degradation process for diatrizoate, a highly recalcitrant ICM, by using biogenic Pd nanoparticles as free suspension or immobilized in polyvinylidene fluoride (PVDF) and polysulfone (PSf) membranes. As measured by HPLC-UV, the removal of 20 mg L⁻¹ diatrizoate by a 10 mg L⁻¹ Pd suspension was completed after 4 h at a pH of 10. LC-MS analysis provided evidence for the sequential hydrodeiodination of diatrizoate. Pd did not lose its activity after incorporation in the PVDF and PSf matrix and the highest activity (k_cat = 30.0 ± 0.4 h⁻¹ L g⁻¹ Pd) was obtained with a casting solution of 10% PSf and 500 mg L⁻¹ Pd. Subsequently, water containing 20 mg L⁻¹ diatrizoate was treated in a membrane contactor, in which the water was supplied at one side of the membrane while hydrogen was provided at the other side. In a fed batch configuration, a removal efficiency of 77% after a time period of 48 h was obtained. This work showed that membrane contactors with encapsulated biogenic nanoparticles can be instrumental for treatment of water contaminated with diatrizoate.     

The removal of pharmaceuticals, personal care products, endocrine disruptors and illicit drugs during wastewater treatment and its impact on the quality of receiving waters

A 5-month monitoring program was undertaken in South Wales in the UK to determine the fate of 55 pharmaceuticals, personal care products, endocrine disruptors and illicit drugs (PPCPs) in two contrasting wastewater plants utilising two different wastewater treatment technologies: activated sludge and trickling filter beds. The impact of treated wastewater effluent on the quality of receiving waters was also assessed.PPCPs were found to be present at high loads reaching 10 kg day⁻¹ in the raw sewage. Concentrations of PPCPs in raw sewage were found to correlate with their usage/consumption patterns in Wales and their metabolism. The efficiency of the removal of PPCPs was found to be strongly dependent on the technology implemented in the wastewater treatment plant (WWTP). In general, the WWTP utilising trickling filter beds resulted in, on average, less than 70% removal of all 55 PPCPs studied, while the WWTP utilising activated sludge treatment gave a much higher removal efficiency of over 85%. The monitoring programme revealed that treated wastewater effluents were the main contributors to PPCPs concentrations (up to 3 kg of PPCPs day⁻¹) in the rivers studied. Bearing in mind that in the cases examined here the WWTP effluents were also major contributors to rivers' flows (dilution factor for the studied rivers did not exceed 23 times) the effect of WWTP effluent on the quality of river water is significant and cannot be underestimated.     

Microorganisms and particles in AHU systems: Measurement and analysis

In this study, for better understanding the practical removal effect of air handling unit (AHU) system on airborne microorganisms (including bacteria and fungus) and particles and microbial growth, the samples of microorganisms and particles in 10 air handling unit (AHU) systems including fan coils and indoor air were collected and analyzed in air and component surfaces of such systems in two large public buildings. It is found that the removal efficiency is of the highest for bacteria 73.9% and the lowest for particles (0.5–2 μm) 24.4%. The surface concentration of equipment bacteria is 29 CFU/cm² and fungi 137 CFU/cm². Five of 10 systems have higher fungi concentrations on air intake than that on diffuser. The results also show that the central air supply system with common components (e.g., pre-filter and bag filter) has difficulty to achieve/maintain good performance once microorganisms and particles exist, especially for particle size D ≤ 3.3 μm. The size distribution has large influence on removal efficiency. The microbial growth on surfaces of duct and equipment was noticed and may be transferred into indoor air. This will decrease the indoor air quality and lead to adverse health effect.     

Accuracy and precision of Legionella isolation by US laboratories in the ELITE program pilot study

A pilot study for the Environmental Legionella Isolation Techniques Evaluation (ELITE) Program, a proficiency testing scheme for US laboratories that culture Legionella from environmental samples, was conducted September 1, 2008 through March 31, 2009. Participants (n = 20) processed panels consisting of six sample types: pure and mixed positive, pure and mixed negative, pure and mixed variable. The majority (93%) of all samples (n = 286) were correctly characterized, with 88.5% of samples positive for Legionella and 100% of negative samples identified correctly. Variable samples were incorrectly identified as negative in 36.9% of reports. For all samples reported positive (n = 128), participants underestimated the cfu/ml by a mean of 1.25 logs with standard deviation of 0.78 logs, standard error of 0.07 logs, and a range of 3.57 logs compared to the CDC re-test value. Centering results around the interlaboratory mean yielded a standard deviation of 0.65 logs, standard error of 0.06 logs, and a range of 3.22 logs. Sampling protocol, treatment regimen, culture procedure, and laboratory experience did not significantly affect the accuracy or precision of reported concentrations. Qualitative and quantitative results from the ELITE pilot study were similar to reports from a corresponding proficiency testing scheme available in the European Union, indicating these results are probably valid for most environmental laboratories worldwide. The large enumeration error observed suggests that the need for remediation of a water system should not be determined solely by the concentration of Legionella observed in a sample since that value is likely to underestimate the true level of contamination.     

Occurrence and removal of N-nitrosamines in wastewater treatment plants

The presence of nitrosamines in wastewater might pose a risk to water resources even in countries where chlorination or chloramination are hardly used for water disinfection. We studied the variation of concentrations and removal efficiencies of eight N-nitrosamines among 21 full-scale sewage treatment plants (STPs) in Switzerland and temporal variations at one of these plants. N-nitrosodimethylamine (NDMA) was the predominant compound in STP primary effluents with median concentrations in the range of 5-20 ng/L, but peak concentrations up to 1 μg/L. N-nitrosomorpholine (NMOR) was abundant in all plants at concentrations of 5-30 ng/L, other nitrosamines occurred at a lower number of plants at similar levels. From concentrations in urine samples and domestic wastewater we estimated that human excretion accounted for levels of <5 ng/L of NDMA and <1 ng/L of the other nitrosamines in municipal wastewater, additional domestic sources for <5 ng/L of NMOR. Levels above this domestic background are probably caused by industrial or commercial discharges, which results in highly variable concentrations in sewage. Aqueous removal efficiencies in activated sludge treatment were in general above 40% for NMOR and above 60% for the other nitrosamines, but could be lower if concentrations were below 8-15 ng/L in primary effluent. We hypothesize that substrate competition in the cometabolic degradation explains the occurrence of such threshold concentrations. An additional sand filtration step resulted in a further removal of nitrosamines from secondary effluents even at low concentrations. Concentrations released to surface waters were largely below 10 ng/L, suggesting a low impact on Swiss water resources and drinking water generation considering the generally high environmental dilution and possible degradation. However, local impacts in case a larger fraction of wastewater is present cannot be ruled out.     

UV inactivation and resistance of rotavirus evaluated by integrated cell culture and real-time RT-PCR assay

Rotaviruses are double-stranded RNA viruses which are among the most resistant waterborne enteric viruses to UV disinfection. An integrated cell culture and real-time RT-PCR (ICC real-time RT-PCR) assay was developed to detect the infectivity of rotaviruses in water, which uses real-time RT-PCR to detect RNA produced by infectious rotaviruses during replication in host cells. Detection of rotaviral RNA in host cells provides direct evidence of the presence of infectious rotavirus rather than just the presence of rotavirus RNA. Using this newly developed method, the inactivation and resistance of rotavirus to UV treatments at various doses was evaluated. With an initial concentration of 2 × 10⁴ PFU/ml simian rotavirus (SA11), a first-order linear relationship was obtained at UV dose range of 0-120 mJ cm⁻², and the inactivation rate constant was estimated to be 0.0343 cm² mJ⁻¹ (R² = 0.966). The dose-inactivation curve tailed off and reached plateau as the UV dose increased from 120 to 360 mJ cm⁻², indicating resistance phenomena of sub-populations of SA11 at very high UV doses. A maximal reduction of 4.8 log₁₀ was observed. Through parallel comparison with traditional culture assay, the ICC real-time RT-PCR method demonstrated more effective, sensitive and faster infectivity detection of rotavirus and, the results reveal that rotaviruses are more resistant to UV irradiation than previously reported with traditional cell culture assays.     

Influence of initial pesticide concentrations and plant population density on dimethomorph toxicity and removal by two duckweed species

Aquatic plants take up, transform and sequester organic contaminants and may therefore be used in phytoremediation for the removal of pollutants from wastewaters. A better understanding of factors affecting the rate of contaminant uptake by aquatic plants is needed to improve engineered systems for removal of pollutants from wastewaters. This work focused on the influence of initial concentrations of pesticide and population density of plants on toxicity and uptake of the fungicide dimethomorph by two duckweed species. An increased sensitivity to dimethomorph was observed with increasing duckweed population density. Less light, due to crowding, may explain this higher sensitivity and reduced removal rate. A positive relationship was also found between toxicity or contaminant uptake and initial pesticide concentration with a maximal removal of 41 and 26 µg g^− 1 fresh weight of dimethomorph (at 600 µg L^− 1 of dimethomorph and an initial density of 0.10 g E-flask^− 1) by Lemna minor and Spirodela polyrhiza, respectively. This research also indicated that these aquatic plants can efficiently eliminate organic contaminants and may ultimately serve as phytoremediation agents in the natural environment.     

Comparison of electrocoagulation and chemical coagulation pretreatment for enhanced virus removal using microfiltration membranes

This research studied virus removal by iron electrocoagulation (EC) followed by microfiltration (MF) in water treatment using the MS2 bacteriophage as a tracer virus. In the absence of EC, MF alone achieved less than a 0.5-log removal of MS2 virus, but, as the iron-coagulant dosage increased, the log virus removal increased dramatically. More than 4-log virus removal, as required by the Surface Water Treatment Rule, was achieved with 6-9 mg/L Fe³⁺. The experimental data indicated that at lower iron dosages and pH (<∼8 mg Fe/L and pH 6.3 and 7.3) negatively charged MS2 viruses first adsorbed onto the positively charged iron hydroxide floc particles before being removed by MF. At higher iron dosages and pH (>∼9 mg Fe/L and pH 8.3), virus removal was attributed predominantly to enmeshment and subsequent removal by MF. Additionally, the experimental data showed no obvious influence of ionic strength in the natural water range of 10⁻⁷-10⁻² M on MS2 virus removal by EC-MF. Finally, EC pretreatment significantly outperformed chemical coagulation pretreatment for virus removal. The proposed mechanism for this improved performance by EC is that locally higher iron and virus concentrations and locally lower pH near the anode improved MS2 enmeshment by iron flocs as well as adsorption of MS2 viruses onto the iron floc particles.