Publications by authors named "Simon Toze"

25 Publications

  • Page 1 of 1

Virological Characterization of Roof-Harvested Rainwater of Densely Urbanized Low-Income Region.

Food Environ Virol 2021 Sep 29;13(3):412-420. Epub 2021 Jun 29.

Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute (IOC), Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, RJ, Brazil.

Roof-harvested rainwater (RHRW) is considered relatively clean water, even though the possible presence of pathogens in the water may pose human health risks. In this study, we investigated the occurrence of enteric viruses in the first flush (10 mm) of RHRW from a densely populated and low-income urbanized region of Rio de Janeiro. One hundred samples (5 L) were collected from 10 rainfall events between April 2015 and March 2017. RNA and DNA viruses were concentrated using the skimmed milk flocculation method and analyzed using the TaqMan® quantitative RT-qPCR and qPCR. Human adenoviruses, noroviruses, rotaviruses A, and avian parvoviruses were detected in 54%, 31%, 12%, and 12% of the positive samples. JC polyomavirus, also targeted, was not detected. Virus concentrations ranged from 1.09 × 10 to 2.58 × 10 genome copies/Liter (GC/L). Partial nucleotide sequence confirmed the presence of HAdV type 41, norovirus genotype GII.4, and avian parvovirus 1. The results suggest that the first flush diversion devices may not adequately remove enteric virus from the rainwater. Additional treatment of RHRW is required to mitigate potential health risks from potable use of captured water.
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http://dx.doi.org/10.1007/s12560-021-09484-yDOI Listing
September 2021

Comparative decay of culturable faecal indicator bacteria, microbial source tracking marker genes, and enteric pathogens in laboratory microcosms that mimic a sub-tropical environment.

Sci Total Environ 2021 Jan 9;751:141475. Epub 2020 Aug 9.

Department of Soil, Water, and Climate, and the BioTechnology Institute, University of Minnesota, St. Paul, MN 55108, USA.

Enteric pathogens can be present in drinking water catchments due to several point and non-point sources of faecal contamination. Pathogen and contaminant signatures will decay due to environmental stresses, such as temperature, Ultra Violet (UV) radiation, salinity, and predation. In this study, we determined the decay of the culturable faecal indicator bacterium (FIB) Escherichia coli (E. coli), two sewage-associated marker genes (Bacteroides HF183 and crAssphage CPQ_056), and enteric pathogens (Campylobacter spp., human adenovirus 40/41, and Cryptosporidium parvum) in two freshwater laboratory microcosms using culture-based, quantitative PCR (qPCR) and vital dye (determine the fraction of viable Cryptosporidium oocysts) assays. Freshwater samples from the Lake Wappa and Lake Wivenhoe (Australia) were seeded with untreated sewage and C. parvum oocysts, and their declining concentrations were measured over a 28-day period. Moreover, 16S rRNA amplicon sequencing was also undertaken to determine the change/shift in sewage-associated bacterial communities using SourceTracker. Overall, culturable E. coli and the HF183 marker gene decayed significantly (p < 0.05) faster than did the qPCR measured enteric pathogens suggesting that the absence of culturable FIB or qPCR HF183 in water samples may not indicate the absence of pathogens. The decay of crAssphage was similar to that of HAdV 40/41 and other pathogens tested, suggesting crAssphage may be a better surrogate for enteric viruses in sub-tropical catchment waters. The decay rates were greater at 25 °C compared to 15 °C, suggesting that FIB and pathogens persist longer in the winter season compared to summer. Overall decay rates of the tested microorganisms in this microcosm study suggest that sub-tropical conditions, especially temperature, have a negative impact on the persistence of tested microorganisms. Sewage-associated bacterial communities also showed similar patterns. Based on the results, which showed differences in simulated summer and winter temperatures for pathogen decay, corresponding management options and treatment need to be adjusted accordingly to minimize human health risks effectively.
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http://dx.doi.org/10.1016/j.scitotenv.2020.141475DOI Listing
January 2021

A review on microbial contaminants in stormwater runoff and outfalls: Potential health risks and mitigation strategies.

Sci Total Environ 2019 Nov 5;692:1304-1321. Epub 2019 Jul 5.

CSIRO Land and Water, Waite Laboratories, Waite Rd., Urrbrae, SA 5064, Australia.

Demands on global water supplies are increasing in response to the need to provide more food, water, and energy for a rapidly growing population. These water stressors are exacerbated by climate change, as well as the growth and urbanisation of industry and commerce. Consequently, urban water authorities around the globe are exploring alternative water sources to meet ever-increasing demands. These alternative sources are primarily treated sewage, stormwater, and groundwater. Stormwater including roof-harvested rainwater has been considered as an alternative water source for both potable and non-potable uses. One of the most significant issues concerning alternative water reuse is the public health risk associated with chemical and microbial contaminants. Several studies to date have quantified fecal indicators and pathogens in stormwater. Microbial source tracking (MST) approaches have also been used to determine the sources of fecal contamination in stormwater and receiving waters. This review paper summarizes occurrence and concentrations of fecal indicators, pathogens, and MST marker genes in urban stormwater. A section of the review highlights the removal of fecal indicators and pathogens through water sensitive urban design (WSUD) or Best Management Practices (BMPs). We also discuss approaches for assessing and mitigating health risks associated with stormwater, including a summary of existing quantitative microbial risk assessment (QMRA) models for potable and non-potable reuse of stormwater. Finally, the most critical research gaps are identified for formulating risk management strategies.
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http://dx.doi.org/10.1016/j.scitotenv.2019.07.055DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7126443PMC
November 2019

Optimization of sampling strategy to determine pathogen removal efficacy of activated sludge treatment plant.

Environ Sci Pollut Res Int 2017 Aug 28;24(23):19001-19010. Epub 2017 Jun 28.

CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Brisbane, 4102, Australia.

Large-scale wastewater schemes rely on multi-barrier approach for the production of safe and sustainable recycled water. In multi-barrier wastewater reclamation systems, conventional activated sludge process (ASP) often constitutes a major initial treatment step. The main aim of this research was to determine most appropriate sampling approach to establish pathogen removal efficacy of ASP. The results suggest that ASP is capable of reducing human adenovirus (HAdV) and polyomavirus (HPyV) by up to 3 log. The virus removal data suggests that HAdV removal is comparable to somatic bacteriophage belonging to Microviridae family. Due to the high removal of Escherichia coli (>3 log) and very poor correlation with the enteric virus, it is not recommended that E. coli be used as a surrogate for enteric virus removal. The results also demonstrated no statistically significant differences (t test, P > 0.05) in calculated log removal values (LRVs) for HAdV, HPyV, and Microviridae from samples collected on hydraulic retention time (HRT) or simultaneous paired samples collected for influent and effluent. This indicates that a more practical approach of simultaneous sampling for influent and effluent could be used to determine pathogen removal efficiency of ASP. The results also suggest that a minimum of 10, preferably 20 samples, are required to fully capture variability in the removal of virus. In order to cover for the potential seasonal prevalence of viruses such as norovirus and rotavirus, sampling should be spread across all seasons.
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http://dx.doi.org/10.1007/s11356-017-9557-5DOI Listing
August 2017

Quantification of hookworm ova from wastewater matrices using quantitative PCR.

J Environ Sci (China) 2017 Jul 24;57:231-237. Epub 2017 Feb 24.

CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Qld 4102, Australia; School of Public Health, The University of Queensland, Herston Road, Qld 4006, Australia.

A quantitative PCR (qPCR) assay was used to quantify Ancylostoma caninum ova in wastewater and sludge samples. We estimated the average gene copy numbers for a single ovum using a mixed population of ova. The average gene copy numbers derived from the mixed population were used to estimate numbers of hookworm ova in A. caninum seeded and unseeded wastewater and sludge samples. The newly developed qPCR assay estimated an average of 3.7×10 gene copies per ovum, which was then validated by seeding known numbers of hookworm ova into treated wastewater. The qPCR estimated an average of (1.1±0.1), (8.6±2.9) and (67.3±10.4) ova for treated wastewater that was seeded with (1±0), (10±2) and (100±21) ova, respectively. The further application of the qPCR assay for the quantification of A. caninum ova was determined by seeding a known numbers of ova into the wastewater matrices. The qPCR results indicated that 50%, 90% and 67% of treated wastewater (1L), raw wastewater (1L) and sludge (~4g) samples had variable numbers of A. caninum gene copies. After conversion of the qPCR estimated gene copy numbers to ova for treated wastewater, raw wastewater, and sludge samples, had an average of 0.02, 1.24 and 67 ova, respectively. The result of this study indicated that qPCR can be used for the quantification of hookworm ova from wastewater and sludge samples; however, caution is advised in interpreting qPCR generated data for health risk assessment.
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http://dx.doi.org/10.1016/j.jes.2016.10.019DOI Listing
July 2017

Comparative prevalence of Escherichia coli carrying virulence genes and class 1 and 2 integrons in sub-tropical and cool temperate freshwater.

Environ Sci Pollut Res Int 2017 Aug 21;24(22):18263-18272. Epub 2017 Jun 21.

CSIRO Land and Water Flagship, Ecosciences Precinct, 41 Boggo Road, Dutton Park, Brisbane, QLD, 4102, Australia.

Aquatic environments are now recognized secondary habitat of potentially pathogenic Escherichia coli. In this study, PCR-based analyses were used to determine the phylogenetic composition and frequency of occurrence of eight clinically significant virulence genes (VGs) in E. coli isolates from sub-tropical Brisbane and cool temperate Tasmania freshwater in Australia. In Brisbane, non-commensal E. coli isolates belonging to the B2 and D phylogenetic group were dominant (72%). A significantly higher number (P < 0.05) of E. coli carrying VGs were detected in the sub-tropical freshwaters compared to the cool temperate water. Furthermore, diarrheagenic pathotype (EHEC) was also observed in the sub-tropical freshwater. The genes east1 and eaeA were significantly more common (P < 0.00001) than other VGs. The eaeA gene which codes for intimin protein along with toxin genes east1, stx , stx , and LT1 were mostly detected in phylogenetic groups B2 and D. The ANOVA results also suggested a statistically significant difference (P < 0.016) between the VGs carried by phylogenetic groups B2 and D. Class 1 integrase (intl1) and class 2 integrase (intl2) genes were detected in 38 (24.83%) and 23 (15.03%) of E. coli isolates, respectively. The Gretna site (Tasmania) with known fecal input from bovine and ovine sources had the highest number of E. coli carrying intl1 (29%) and intl2 (13%) genes. In addition, class 2 integron was more commonly detected in the phylogenetic group B2. The results of this study highlight the need to better understand sources and reasons for the high prevalence of E. coli carrying clinically significant VGs in a sub-tropical environment and its public health implications.
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http://dx.doi.org/10.1007/s11356-017-9497-0DOI Listing
August 2017

Human health risks for Legionella and Mycobacterium avium complex (MAC) from potable and non-potable uses of roof-harvested rainwater.

Water Res 2017 08 5;119:288-303. Epub 2017 Apr 5.

Drexel University, 3141 Chestnut Street, Philadelphia, PA 19104, USA.

A quantitative microbial risk assessment (QMRA) of opportunistic pathogens Legionella pneumophila (LP) and Mycobacterium avium complex (MAC) was undertaken for various uses of roof-harvested rainwater (RHRW) reported in Queensland, Australia to identify appropriate usages and guide risk management practices. Risks from inhalation of aerosols due to showering, swimming in pools topped up with RHRW, use of a garden hose, car washing, and toilet flushing with RHRW were considered for LP while both ingestion (drinking, produce consumption, and accidental ingestion from various activities) and inhalation risks were considered for MAC. The drinking water route of exposure presented the greatest risks due to cervical lymphadenitis and disseminated infection health endpoints for children and immune-compromised populations, respectively. It is therefore not recommended that these populations consume untreated rainwater. LP risks were up to 6 orders of magnitude higher than MAC risks for the inhalation route of exposure for all scenarios. Both inhalation and ingestion QMRA simulations support that while drinking, showering, and garden hosing with RHRW may present the highest risks, car washing and clothes washing could constitute appropriate uses of RHRW for all populations, and toilet flushing and consumption of lettuce irrigation with RHRW would be appropriate for non- immune-compromised populations.
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http://dx.doi.org/10.1016/j.watres.2017.04.004DOI Listing
August 2017

Seasonal Assessment of Opportunistic Premise Plumbing Pathogens in Roof-Harvested Rainwater Tanks.

Environ Sci Technol 2017 02 26;51(3):1742-1753. Epub 2017 Jan 26.

Drexel University , 3141 Chestnut Street, Philadelphia, Pennsylvania 19104, United States.

A seasonal study on the occurrence of six opportunistic premise plumbing pathogens (OPPPs) in 24 roof-harvested rainwater (RHRW) tanks repeatedly sampled over six monthly sampling events (n = 144) from August 2015 to March 2016 was conducted using quantitative qPCR. Fecal indicator bacteria (FIB) Escherichia coli (E. coli) and Enterococcus spp. were enumerated using culture-based methods. All tank water samples over the six events were positive for at least one OPPP (Legionella spp., Legionella pneumophila, Mycobacterium avium, Mycobacterium intracellulare, Pseudmonas aeruginosa, or Acanthamoeba spp.) during the entire course of the study. FIB were positively but weakly correlated with P. aeruginosa (E. coli vs P. aeruginosa τ = 0.090, p = 0.027; Enterococcus spp. vs P. aeruginosa τ = 0.126, p = 0.002), but not the other OPPPs. FIBs were more prevalent during the wet season than the dry season, and L. pneumophila was only observed during the wet season. However, concentrations of Legionella spp., M. intracellulare, Acanthamoeba spp., and M. avium peaked during the dry season. Correlations were assessed between FIB and OPPPs with meteorological variables, and it was determined that P. aeruginosa was the only OPPP positively associated with an increased antecedent dry period, suggesting stagnation time may play a role for the occurrence of this OPPP in tank water. Infection risks may exceed commonly cited benchmarks for uses reported in the rainwater usage survey such as pool top-up, and warrant further exploration through quantitative microbial risk assessment (QMRA).
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http://dx.doi.org/10.1021/acs.est.6b04814DOI Listing
February 2017

Amplicon-based profiling of bacteria in raw and secondary treated wastewater from treatment plants across Australia.

Appl Microbiol Biotechnol 2017 Feb 12;101(3):1253-1266. Epub 2016 Nov 12.

CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Brisbane, QLD, 4102, Australia.

In this study, we investigated the use of Illumina high-throughput sequencing of 16S ribosomal RNA (rRNA) amplicons to explore microbial diversity and community structure in raw and secondary treated wastewater (WW) samples from four municipal wastewater treatment plants (WWTPs A-D) across Australia. Sequence reads were analyzed to determine the abundance and diversity of bacterial communities in raw and secondary treated WW samples across the four WWTPs. In addition, sequence reads were also characterized to phenotypic features and to estimate the abundance of potential pathogenic bacterial genera and antibiotic-resistant genes in total bacterial communities. The mean coverage, Shannon diversity index, observed richness (S ), and abundance-based coverage estimate (ACE) of richness for raw and secondary treated WW samples did not differ significantly (P > 0.05) among the four WWTPs examined. Generally, raw and secondary treated WW samples were dominated by members of the genera Pseudomonas, Arcobacter, and Bacteroides. Evaluation of source contributions to secondary treated WW, done using SourceTracker, revealed that 8.80-61.4% of the bacterial communities in secondary treated WW samples were attributed to raw WW. Twenty-five bacterial genera were classified as containing potential bacterial pathogens. The abundance of potentially pathogenic genera in raw WW samples was higher than that found in secondary treated WW samples. Among the pathogenic genera identified, Pseudomonas and Arcobacter had the greatest percentage of the sequence reads. The abundances of antibiotic resistance genes were generally low (<0.5%), except for genes encoding ABC transporters, which accounted for approximately 3% of inferred genes. These findings provided a comprehensive profile of bacterial communities, including potential bacterial pathogens and antibiotic-resistant genes, in raw and secondary treated WW samples from four WWTPs across Australia and demonstrated that Illumina high-throughput sequencing can be an alternative approach for monitoring WW quality in order to protect environmental and human health.
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http://dx.doi.org/10.1007/s00253-016-7959-9DOI Listing
February 2017

Attachment and Detachment Behavior of Human Adenovirus and Surrogates in Fine Granular Limestone Aquifer Material.

J Environ Qual 2015 Sep;44(5):1392-401

The transport of human adenovirus, nanoparticles, and PRD1 and MS2 bacteriophages was tested in fine granular limestone aquifer material taken from a borehole at a managed aquifer recharge site in Adelaide, South Australia. Comparison of transport and removal of virus surrogates with the pathogenic virus is necessary to understand the differences between the virus and surrogate. Because experiments using pathogenic viruses cannot be done in the field, laboratory tests using flow-through soil columns were used. Results show that PRD1 is the most appropriate surrogate for adenovirus in an aquifer dominated by calcite material but not under high ionic strength or high pH conditions. It was also found that straining due to size and the charge of the colloid were not dominant removal mechanisms in this system. Implications of this study indicate that a certain surrogate may not represent a specific pathogen solely based on similar size, morphology, and/or surface charge. Moreover, if a particular surrogate is representative of a pathogen in one aquifer system, it may not be the most appropriate surrogate in another porous media system. This was apparent in the inferior performance of MS2 as a surrogate, which is commonly used in virus transport studies.
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http://dx.doi.org/10.2134/jeq2015.01.0052DOI Listing
September 2015

Assessment of Genetic Markers for Tracking the Sources of Human Wastewater Associated Escherichia coli in Environmental Waters.

Environ Sci Technol 2015 Aug 22;49(15):9341-6. Epub 2015 Jul 22.

†CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Brisbane, Queensland 4102, Australia.

In this study, we have evaluated the performance characteristics (host-specificity and -sensitivity) of four human wastewater-associated Escherichia coli (E. coli) genetic markers (H8, H12, H14, and H24) in 10 target (human) and nontarget (cat, cattle, deer, dog, emu, goat, horse, kangaroo, and possum) host groups in Southeast Queensland, Australia. The overall host-sensitivity values of the tested markers in human wastewater samples were 1.0 (all human wastewater samples contained the E. coli genetic markers). The overall host-specificity values of these markers to differentiate between human and animal host groups were 0.94, 0.85, 0.72, and 0.57 for H8, H12, H24, and H14, respectively. Based on the higher host-specificity values, H8 and H12 markers were chosen for a validation environmental study. The prevalence of the H8 and H12 markers was determined among human wastewater E. coli isolates collected from a wastewater treatment plant (WWTP). Among the 97 isolates tested, 44 (45%) and 14 (14%) were positive for the H8 and H12 markers, respectively. A total of 307 E. coli isolates were tested from environmental water samples collected in Brisbane, of which 7% and 20% were also positive for the H8 and H12 markers, respectively. Based on our results, we recommend that these markers could be useful when it is important to identify the source(s) of E. coli (whether they originated from human wastewater or not) in environmental waters.
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http://dx.doi.org/10.1021/acs.est.5b02163DOI Listing
August 2015

Evaluating two infiltration gallery designs for managed aquifer recharge using secondary treated wastewater.

J Environ Manage 2013 Mar 4;117:115-20. Epub 2013 Feb 4.

CSIRO Water for a Healthy Country Flagship, CSIRO Centre for Environment and Life Sciences, Private Bag No. 5, PO Wembley, Western Australia 6913, Australia.

As managed aquifer recharge (MAR) becomes increasingly considered for augmenting water-sensitive urban areas, fundamental knowledge of the achievable scale, longevity and maintenance requirements of different options will become paramount. This paper reports on a 39 month pilot scale MAR scheme that infiltrated secondary treated wastewater through unsaturated sand into a limestone and sand aquifer. Two types of infiltration gallery were constructed to compare their hydraulic performance, one using crushed, graded gravel, the other using an engineered leach drain system (Atlantis Leach System(®)). Both galleries received 25 kL of nutrient-rich, secondary treated wastewater per day. The Atlantis gallery successfully infiltrated 17 ML of treated wastewater over three years. The slotted distribution pipe in the gravel gallery became clogged with plant roots after operating for one year. The infiltration capacity of the gravel gallery could not be restored despite high pressure cleaning, thus it was replaced with an Atlantis system. Reduction in the infiltration capacity of the Atlantis system was only observed when inflow was increased by about 3 fold for two months. The performance of the Atlantis system suggests it is superior to the gravel gallery, requiring less maintenance within at least the time frame of this study. The results from a bromide tracer test revealed a minimum transport time of 3.7 days for the recharged water to reach the water table below 9 m of sand and limestone. This set a limit on the time available for attenuation by natural treatment within the unsaturated zone before it recharged groundwater.
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http://dx.doi.org/10.1016/j.jenvman.2012.12.018DOI Listing
March 2013

Urban stormwater harvesting and reuse: a probe into the chemical, toxicology and microbiological contaminants in water quality.

Environ Monit Assess 2013 Aug 21;185(8):6645-52. Epub 2012 Dec 21.

CSIRO Land and Water, Ecosciences Precinct, Boggo Rd, Dutton Park, Queensland, 4102, Australia.

Stormwater is one of the last major untapped urban water resources that can be exploited as an alternative water source in Australia. The information in the current Australian Guidelines for Water Recycling relating to stormwater harvesting and reuse only emphasises on a limited number of stormwater quality parameters. In order to supply stormwater as a source for higher value end-uses, a more comprehensive assessment on the potential public health risks has to be undertaken. Owing to the stochastic variations in rainfall, catchment hydrology and also the types of non-point pollution sources that can provide contaminants relating to different anthropogenic activities and catchment land uses, the characterisation of public health risks in stormwater is complex, tedious and not always possible through the conventional detection and analytical methods. In this study, a holistic approach was undertaken to assess the potential public health risks in urban stormwater samples from a medium-density residential catchment. A combined chemical-toxicological assessment was used to characterise the potential health risks arising from chemical contaminants, while a combination of standard culture methods and quantitative polymerase chain reaction (qPCR) methods was used for detection and quantification of faecal indicator bacteria (FIB) and pathogens in urban stormwater. Results showed that the concentration of chemical contaminants and associated toxicity were relatively low when benchmarked against other alternative water sources such as recycled wastewater. However, the concentrations of heavy metals particularly cadmium and lead have exceeded the Australian guideline values, indicating potential public health risks. Also, high numbers of FIB were detected in urban stormwater samples obtained from wet weather events. In addition, qPCR detection of human-related pathogens suggested there are frequent sewage ingressions into the urban stormwater runoff during wet weather events. Further water quality monitoring study will be conducted at different contrasting urban catchments in order to undertake a more comprehensive public health risk assessment for urban stormwater.
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http://dx.doi.org/10.1007/s10661-012-3053-7DOI Listing
August 2013

Sensitive detection of human adenovirus from small volume of primary wastewater samples by quantitative PCR.

J Virol Methods 2013 Feb 23;187(2):395-400. Epub 2012 Nov 23.

CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Qld 4102, Australia.

An accurate quantitative detection of enteric viruses from the primary wastewater requires, sample concentration followed by extraction of nucleic acid with high purity. A highly efficient and sensitive method was developed for the concentration and quantitative detection of human adenovirus (HAdv) from wastewater samples. The two-step method which combines concentration of virus from 10 mL sample with centrifugal filters followed by extraction and purification of DNA with commercially available nucleic acid extraction kit resulted in high purity DNA for downstream quantitative PCR (qPCR). The results obtained on analytical sensitivities of five commercial nucleic acid extraction kits show that they differ in their ability for DNA yield and purity. Nevertheless, despite variable analytical sensitivities extracted nucleic acid was found to be relatively PCR inhibition free. The genomic copy numbers of HAdv detected from the same concentrated wastewater sample were significantly higher (P<0.01) when Qiagen Blood and Tissue kit (1.54×10(6) L(-1)) was used as compared to Mo-Bio PowerSoil kit (5.30×10(5) L(-1)) which suggests that the nucleic acid extraction kit can influence the sensitivity of qPCR assays. The method developed in this study is simple, rapid, sensitive, and can be applicable for the qPCR detection of adenovirus and other DNA virus in wastewater.
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http://dx.doi.org/10.1016/j.jviromet.2012.11.002DOI Listing
February 2013

Occurrence of virulence genes associated with Diarrheagenic pathotypes in Escherichia coli isolates from surface water.

Appl Environ Microbiol 2013 Jan 2;79(1):328-35. Epub 2012 Nov 2.

CSIRO Land and Water, Ecosciences Precinct, Queensland, Australia.

Escherichia coli isolates (n = 300) collected from six sites in subtropical Brisbane, Australia, prior to and after storm events were tested for the presence of 11 virulence genes (VGs) specific to diarrheagenic pathotypes. The presence of eaeA, stx(1), stx(2), and ehxA genes specific for the enterohemorrhagic E. coli (EHEC) pathotype was detected in 56%, 6%, 10%, and 13% of isolates, respectively. The VGs astA (69%) and aggR (29%), carried by enteroaggregative (EAEC) pathotypes, were frequently detected in E. coli isolates. The enteropathogenic E. coli (EPEC) gene bfp was detected in 24% of isolates. In addition, enteroinvasive E. coli (EIEC) VG ipaH was also detected in 14% of isolates. During dry periods, isolates belonging to the EAEC pathotype were most commonly detected (23%), followed by EHEC (11%) and EPEC (11%). Conversely, a more uniform prevalence of pathotypes, EPEC (14%), EAEC (12%), EIEC (10%), EHEC (7%), and ETEC (7%), was observed after the storm events. The results of this study highlight the widespread occurrence of potentially diarrheagenic pathotypes in the urban aquatic ecosystems. While the presence of VGs in E. coli isolates alone is insufficient to determine pathogenicity, the presence of diarrheagenic E. coli pathotypes in high frequency after the storm events could lead to increased health risks if untreated storm water were to be used for nonpotable purposes and recreational activities.
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http://dx.doi.org/10.1128/AEM.02888-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3536071PMC
January 2013

Water quality assessment using the AREc32 reporter gene assay indicative of the oxidative stress response pathway.

J Environ Monit 2012 Nov;14(11):2877-85

National Research Centre for Environmental Toxicology (Entox), The University of Queensland, Brisbane, QLD 4108, Australia.

The reporter gene assay AREc32 is based on the induction of the Nrf2 mediated oxidative stress response pathway in the human breast cancer cell line MCF7, where eight copies of the antioxidant response element (ARE) are linked to a reporter gene encoding for luciferase. The Nrf2-ARE pathway is responsive to many chemicals that cause oxidative stress, among them a large number of pesticides and skin irritants. We adopted and validated the AREc32 bioassay for water quality testing. tert-Butylhydroquinone served as the positive control, phenol as the negative control and other reactive chemicals were assessed for their specificity. An environmentally relevant reference chemical, benzo(a)pyrene was the most potent inducer of all tested chemicals. The concentration causing an induction ratio (IR) of 1.5 (EC(IR1.5)) was chosen as the effect benchmark value. The assay was applied to 21 water samples ranging from sewage to drinking water, including secondary treatment and various tertiary treatment options (ozonation, biologically activated carbon filtration, membrane filtration, reverse osmosis, advanced oxidation, chlorination, chloramination). The samples were enriched by solid phase extraction. In most samples the oxidative stress response was far more sensitive than cytotoxicity. The primary and secondary treated effluent exceeded the effect threshold IR 1.5 at a relative enrichment factor (REF) of 1, i.e., the native samples were active. All tertiary treated samples were less potent and their EC(IR1.5) lay between REF 1 and 10. The Nrf2 pathway was induced at a REF of approximately 10 for surface waters and drinking water, and above this enrichment cytotoxicity took over in most samples and quenched the induction. The blank (ultrapure water run through the sample enrichment process) was cytotoxic at an REF of 100, which is the limit of concentrations range that can be evaluated. Treatment typically decreased both the cytotoxicity and oxidative stress response apart from drinking water treatment where chlorination caused an increase in oxidative stress response, presumably due to the formation of disinfection by-products. This study demonstrates the relevance and applicability of the oxidative stress response pathway for water quality monitoring.
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http://dx.doi.org/10.1039/c2em30506bDOI Listing
November 2012

Managed aquifer recharge of treated wastewater: water quality changes resulting from infiltration through the vadose zone.

Water Res 2011 Nov 3;45(17):5764-72. Epub 2011 Sep 3.

CSIRO Water for a Healthy Country Flagship, CSIRO Centre for Environment and Life Sciences, Wembley, Western Australia, Australia.

Secondary treated wastewater was infiltrated through a 9 m-thick calcareous vadose zone during a 39 month managed aquifer recharge (MAR) field trial to determine potential improvements in the recycled water quality. The water quality improvements of the recycled water were based on changes in the chemistry and microbiology of (i) the recycled water prior to infiltration relative to (ii) groundwater immediately down-gradient from the infiltration gallery. Changes in the average concentrations of several constituents in the recycled water were identified with reductions of 30% for phosphorous, 66% for fluoride, 62% for iron and 51% for total organic carbon when the secondary treated wastewater was infiltrated at an applied rate of 17.5 L per minute with a residence time of approximately four days in the vadose zone and less than two days in the aquifer. Reductions were also noted for oxazepam and temazepam among the pharmaceuticals tested and for a range of microbial pathogens, but reductions were harder to quantify as their magnitudes varied over time. Total nitrogen and carbamazepine persisted in groundwater down-gradient from the infiltration galleries. Infiltration does potentially offer a range of water quality improvements over direct injection to the water table without passage through the unsaturated zone; however, additional treatment options for the non-potable water may still need to be considered, depending on the receiving environment or the end use of the recovered water.
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http://dx.doi.org/10.1016/j.watres.2011.08.058DOI Listing
November 2011

Bioanalytical tools for the evaluation of organic micropollutants during sewage treatment, water recycling and drinking water generation.

Water Res 2011 Aug 7;45(14):4238-47. Epub 2011 Jun 7.

The University of Queensland, National Research Centre for Environmental Toxicology (Entox), 39 Kessels Rd, Brisbane, Qld 4108, Australia.

A bioanalytical test battery was used for monitoring organic micropollutants across an indirect potable reuse scheme testing sites across the complete water cycle from sewage to drinking water to assess the efficacy of different treatment barriers. The indirect potable reuse scheme consists of seven treatment barriers: (1) source control, (2) wastewater treatment plant, (3) microfiltration, (4) reverse osmosis, (5) advanced oxidation, (6) natural environment in a reservoir and (7) drinking water treatment plant. Bioanalytical results provide complementary information to chemical analysis on the sum of micropollutants acting together in mixtures. Six endpoints targeting the groups of chemicals with modes of toxic action of particular relevance for human and environmental health were included in the evaluation: genotoxicity, estrogenicity (endocrine disruption), neurotoxicity, phytotoxicity, dioxin-like activity and non-specific cell toxicity. The toxicity of water samples was expressed as toxic equivalent concentrations (TEQ), a measure that translates the effect of the mixtures of unknown and potentially unidentified chemicals in a water sample to the effect that a known reference compound would cause. For each bioassay a different representative reference compound was selected. In this study, the TEQ concept was applied for the first time to the umuC test indicative of genotoxicity using 4-nitroquinoline as the reference compound for direct genotoxicity and benzo[a]pyrene for genotoxicity after metabolic activation. The TEQ were observed to decrease across the seven treatment barriers in all six selected bioassays. Each bioassay showed a differentiated picture representative for a different group of chemicals and their mixture effect. The TEQ of the samples across the seven barriers were in the same order of magnitude as seen during previous individual studies in wastewater and advanced water treatment plants and reservoirs. For the first time a benchmarking was performed that allows direct comparison of different treatment technologies and covers several orders of magnitude of TEQ from highly contaminated sewage to drinking water with TEQ close or below the limit of detection. Detection limits of the bioassays were decreased in comparison to earlier studies by optimizing sample preparation and test protocols, and were comparable to or lower than the quantification limits of the routine chemical analysis, which allowed monitoring of the presence and removal of micropollutants post Barrier 2 and in drinking water. The results obtained by bioanalytical tools were reproducible, robust and consistent with previous studies assessing the effectiveness of the wastewater and advanced water treatment plants. The results of this study indicate that bioanalytical results expressed as TEQ are useful to assess removal efficiency of micropollutants throughout all treatment steps of water recycling.
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http://dx.doi.org/10.1016/j.watres.2011.05.032DOI Listing
August 2011

Incorporating parameter uncertainty into Quantitative Microbial Risk Assessment (QMRA).

J Water Health 2011 Mar;9(1):10-26

Queensland University of Technology, George Street, Brisbane, QLD 4000, Australia.

Modern statistical models and computational methods can now incorporate uncertainty of the parameters used in Quantitative Microbial Risk Assessments (QMRA). Many QMRAs use Monte Carlo methods, but work from fixed estimates for means, variances and other parameters. We illustrate the ease of estimating all parameters contemporaneously with the risk assessment, incorporating all the parameter uncertainty arising from the experiments from which these parameters are estimated. A Bayesian approach is adopted, using Markov Chain Monte Carlo Gibbs sampling (MCMC) via the freely available software, WinBUGS. The method and its ease of implementation are illustrated by a case study that involves incorporating three disparate datasets into an MCMC framework. The probabilities of infection when the uncertainty associated with parameter estimation is incorporated into a QMRA are shown to be considerably more variable over various dose ranges than the analogous probabilities obtained when constants from the literature are simply 'plugged' in as is done in most QMRAs. Neglecting these sources of uncertainty may lead to erroneous decisions for public health and risk management.
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http://dx.doi.org/10.2166/wh.2010.073DOI Listing
March 2011

Risk assessment of aquifer storage transfer and recovery with urban stormwater for producing water of a potable quality.

J Environ Qual 2010 Nov-Dec;39(6):2029-39

CSIRO Land and Water, Private Bag No 2, Glen Osmond, SA 5064, Australia.

The objective of the Parafield Aquifer Storage Transfer and Recovery research project in South Australia is to determine whether stormwater from an urban catchment that is treated in a constructed wetland and stored in an initially brackish aquifer before recovery can meet potable water standards. The water produced by the stormwater harvesting system, which included a constructed wetland, was found to be near potable quality. Parameters exceeding the drinking water guidelines before recharge included small numbers of fecal indicator bacteria and elevated iron concentrations and associated color. This is the first reported study of a managed aquifer recharge (MAR) scheme to be assessed following the Australian guidelines for MAR. A comprehensive staged approach to assess the risks to human health and the environment of this project has been undertaken, with 12 hazards being assessed. A quantitative microbial risk assessment undertaken on the water recovered from the aquifer indicated that the residual risks posed by the pathogenic hazards were acceptable if further supplementary treatment was included. Residual risks from organic chemicals were also assessed to be low based on an intensive monitoring program. Elevated iron concentrations in the recovered water exceeded the potable water guidelines. Iron concentrations increased after underground storage but would be acceptable after postrecovery aeration treatment. Arsenic concentrations in the recovered water continuously met the guideline concentrations acceptable for potable water supplies. However, the elevated concentration of arsenic in native groundwater and its presence in aquifer minerals suggest that the continuing acceptable residual risk from arsenic requires further evaluation.
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http://dx.doi.org/10.2134/jeq2010.0078DOI Listing
February 2011

Valuing the subsurface pathogen treatment barrier in water recycling via aquifers for drinking supplies.

Water Res 2010 Mar 16;44(6):1841-52. Epub 2009 Dec 16.

CSIRO Water for a Healthy Country, Private Bag No. 2, Glen Osmond, SA 5064, Australia.

A quantitative microbial risk assessment (QMRA) was performed at four managed aquifer recharge (MAR) sites (Australia, South Africa, Belgium, Mexico) where reclaimed wastewater and stormwater is recycled via aquifers for drinking water supplies, using the same risk-based approach that is used for public water supplies. For each of the sites, the aquifer treatment barrier was assessed for its log(10) removal capacity much like for other water treatment technologies. This information was then integrated into a broader risk assessment to determine the human health burden from the four MAR sites. For the Australian and South African cases, managing the aquifer treatment barrier was found to be critical for the schemes to have low risk. For the Belgian case study, the large treatment trains both in terms of pre- and post-aquifer recharge ensures that the risk is always low. In the Mexico case study, the risk was high due to the lack of pre-treatment and the low residence times of the recharge water in the aquifer. A further sensitivity analysis demonstrated that human health risk can be managed if aquifers are integrated into a treatment train to attenuate pathogens. However, reduction in human health disease burden (as measured in disability adjusted life years, DALYs) varied depending upon the number of pathogens in the recharge source water. The beta-Poisson dose response curve used for translating rotavirus and Cryptosporidium numbers into DALYs coupled with their slow environmental decay rates means poor quality injectant leads to aquifers having reduced value to reduce DALYs. For these systems, like the Mexican case study, longer residence times are required to meet their DALYs guideline for drinking water. Nevertheless the results showed that the risks from pathogens can still be reduced and recharging via an aquifer is safer than discharging directly into surface water bodies.
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http://dx.doi.org/10.1016/j.watres.2009.12.008DOI Listing
March 2010

Use of static Quantitative Microbial Risk Assessment to determine pathogen risks in an unconfined carbonate aquifer used for Managed Aquifer Recharge.

Water Res 2010 Feb 27;44(4):1038-49. Epub 2009 Aug 27.

CSIRO Water for a Healthy Country, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia, Brisbane, QLD 4067, Australia.

Managed Aquifer Recharge (MAR) is becoming a mechanism used for recycling treated wastewater and captured urban stormwater and is being used as a treatment barrier to remove contaminants such as pathogens from the recharged water. There is still a need, however, to demonstrate the effectiveness of MAR to reduce any residual risk of pathogens in the recovered water. A MAR research site recharging secondary treated wastewater in an unconfined carbonate aquifer was used in conjunction with a static Quantitative Microbial Risk Assessment (QMRA) to assess the microbial pathogen risk in the recovered water following infiltration and aquifer passage. The research involved undertaking a detailed hydrogeological assessment of the aquifer at the MAR site and determining the decay rates of reference pathogens from an in-situ decay study. These variables along with literature data were then used in the static QMRA which demonstrated that the recovered water at this site did not meet the Australian Guidelines for recycled water when used for differing private green space irrigation scenarios. The results also confirmed the importance of obtaining local hydrogeological data as local heterogeneity can influence of residence time in the aquifer which, in turn, influences the outcomes. The research demonstrated that a static QMRA can be used to determine the residual risk from pathogens in recovered water and showed that it can be a valuable tool in the preliminary design and operation of MAR systems and the incorporation of complementary engineered treatment processes to ensure that there is acceptable health risk from the recovered water.
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http://dx.doi.org/10.1016/j.watres.2009.08.028DOI Listing
February 2010

Human pathogens and their indicators in biosolids: a literature review.

Environ Int 2009 Jan 13;35(1):187-201. Epub 2008 Sep 13.

CSIRO Land and Water, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia, QLD 4067, Australia.

A growing beneficial reuse of biosolids in agriculture has led to concerns about potential contamination of water resources and the food chain. In order to comprehend the potential risks of transmission of diseases to the human population, an advanced quantitative risk assessment is essential. This requires good quantitative data which is currently limited due to the methodological limitations. Consequently, further development and standardization of methodologies for the detection, enumeration and viability assessment of pathogens in biosolids is required. There is a paucity of information on the numbers and survival of enteric virus and protozoan pathogens of concern in biosolids. There is a growing urgency for the identification of more reliable alternative indicators, both index and model microorganisms, which could be used for potential public health risk assessment. In this review, we have summarized reported literature on the numbers and fate of enteric pathogens and indicators in biosolids. The advantages and limitations of the use of conventional and alternative index and model microorganisms for the prediction of pathogen presence in biosolids are also discussed.
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http://dx.doi.org/10.1016/j.envint.2008.07.006DOI Listing
January 2009

Decay of endocrine-disrupting chemicals in aerobic and anoxic groundwater.

Water Res 2008 Feb 7;42(4-5):1133-41. Epub 2007 Sep 7.

CSIRO Land and Water, PMB2, Glen Osmond, SA 5064, Australia.

Biodegradation and adsorption of selected endocrine-disrupting chemicals (EDCs), namely oestrogens (E2 and EE2) and phenolic compounds (BPA, 4-t-OP and 4-n-NP), in aquifer materials was investigated in the laboratory in order to understand the behaviour and fate of these chemicals associated with reclaimed water during managed aquifer recharge. Biodegradation experiments were conducted in microcosms with aquifer material and groundwater mixture, or with aquifer material and effluent mixture in the presence of glucose under both aerobic and anoxic conditions. All five selected compounds were degraded by microorganisms in both types of aquifer material-water mixtures under aerobic and anoxic conditions. Under aerobic conditions, EE2 was found to degrade faster in the aquifer material supplemented with effluent with a half-life of 15 days compared with that of 26 days in the aquifer material and groundwater microcosm. No significant difference between the two aquifer material-water mixtures was found for the other four compounds with half-lives ranging between 0.2 and 4.1 days. Under anoxic conditions, however, little biodegradation was observed for the selected EDCs except for E2, which degraded in the aquifer material in the presence of both water types.
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http://dx.doi.org/10.1016/j.watres.2007.08.029DOI Listing
February 2008

Persistence of biofilm-associated Escherichia coli and Pseudomonas aeruginosa in groundwater and treated effluent in a laboratory model system.

Microbiology (Reading) 2003 Jan;149(Pt 1):47-55

CSIRO Land and Water, Floreat, Western Australia.

This study was based on the hypothesis that groundwater-derived biofilms may provide a reservoir for coliform or pathogenic bacteria as has been observed in drinking water distribution systems. Escherichia coli, labelled with green fluorescent protein, was found to colonize all layers of mixed-population biofilms developed in association with indigenous groundwater micro-organisms in a laboratory-scale reactor. Biofilm-associated E. coli was removed at a slower rate from the reactor flasks than planktonic E. coli under a continuous flow regime. During flow-through of groundwater, planktonic E. coli removal was slower in flasks containing coverslips for enhanced biofilm development compared to a control flask without coverslips. Conversely, during flow-through of treated effluent, planktonic E. coli removal was faster in flasks with coverslips compared to without. Removal of attached E. coli was also fastest in the coverslip-containing flasks with effluent flow-through. This suggests that an increase in available nutrients may reduce E. coli survival potential due to either enhanced competition for nutrients or enhanced antagonism by the indigenous microbial population. Under identical conditions, GFP-labelled Pseudomonas aeruginosa was found to persist in the biofilms for longer than E. coli, most notably when exposed to flow-through of treated effluent. However, prolonged persistence of P. aeruginosa in the effluent could not be attributed to an association with the biofilms. This study has shown that under certain conditions the presence of mixed-population biofilms may limit the survival potential of enteric bacterial pathogens introduced into groundwater.
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http://dx.doi.org/10.1099/mic.0.25938-0DOI Listing
January 2003
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