Biomedical subjects

N J Ashbolt

Publications and source records attributed to N J Ashbolt.

At least 19 recordsLinked to original sources

PubMed2006

Soil inactivation of DNA viruses in septic seepage.

AIMS: To generate field-relevant inactivation data for incorporation into models to predict the likelihood of viral contamination of surface waters by septic seepage. METHODS AND RESULTS: Inactivation rates were determined for PRD1 bacteriophage and Adenovirus 2 in two catchment soils under a range of temperature, moisture and biotic status regimes. Inactivation rates presented for both viruses were significantly different at different temperatures and in different soil types (alpha = 0.05). Soil moisture generally did not significantly affect virus inactivation rate. Biotic status significantly affected inactivation rates of PRD1 in the loam soil but not the clay-loam soil. Adenovirus 2 was inactivated more rapidly in the loam soil than PRD1 bacteriophage. CONCLUSIONS: Virus inactivation rates incorporated into models should be appropriate for the climate/catchment in question with particular regard to soil type and temperature. Given that PRD1 is similar in size to adenoviruses, yet more conservative with regard to inactivation in soil, it may be a useful surrogate in studies of Adenovirus fate and transport. SIGNIFICANCE AND IMPACT OF THE STUDY: A better understanding of the factors that govern virus fate and transport in catchments would facilitate the design of barrier measures to prevent viral contamination of surface waters by septic seepage.

Adenoviridae↗

PubMed2006

Microbial exposure assessment of an urban recreational lake: a case study of the application of new risk-based guidelines.

New WHO and Australian guidelines promote a risk-management approach for minimising exposure to pathogens in recreational waters. Between 2003 and 2005, they were applied to Lake Parramatta (10 ha, 450 ML), a potential recreation site in Sydney, Australia. A three stage approach was developed involving (1) initial suitability assessment using historic data, (2) revised suitability assessment based on new data and (3) characterisation of hazardous (especially wet weather) events. Contrary to the stage 1 suitability classification, stage 2 baseline data indicated that during dry weather the lake had water quality sufficient for primary contact recreation (95th percentiles for enterococci = 19 MPN/100, n = 50) and the major pathogen source was wildfowl. Guideline principles provided a rationale for collecting microbiological and geographic data needed to understand local cycles of lake contamination/recovery. The concept of hazardous events was particularly useful. Studies of stormwater events led us to identify a transition point (> 10 mm rainfall in 24 h) where human-faecal pathogen risks increased and access needed to be controlled. Together baseline and event data yielded operational tools (i.e. event detection methods, action triggers, auditing criteria, remediation priorities) for minimising bather exposure.

Clostridium perfringens↗

PubMed2006

Microbial risk and removal--a utility perspective.

In this paper the results of a sampling programme, undertaken as part of the EU MicroRisk project, are described. This project was undertaken to ascertain the occurrence of pathogens and indicators in the River Thames and their subsequent removal through a treatment works. Appropriate physico-chemical surrogates, as determined by statistical correlation are proposed for the microorganisms identified in the raw water. This study shows that under normal raw water conditions the treatment works is able to remove microbial contamination with a significant margin of safety.

Animals↗

PubMed2006

Pathogen monitoring offers questionable protection against drinking-water risks: a QMRA (quantitative microbial risk analysis) approach to assess management strategies.

Risk mitigation provided by human monitoring and control over a water supply system has been consistently overlooked when estimating pathogen exposure to consumers. The Systems-Actions-Management (SAM) framework lends itself neatly to Quantitative Microbial Risk Assessment (QMRA) as one way to establish this link. The general premise is that an organisational protocol will influence how a human controller behaves, in turn influencing the system performance. For illustrative purposes, the framework was applied to a hypothetical water supply system to quantify the risk reduction offered by routine Cryptosporidium monitoring and the response to oocyst 'detects'. Our findings suggest that infrequent direct pathogen monitoring may provide a negligible risk barrier. The practice of sampling treated water to verify microbiological integrity is also dubious: oocyst densities were largely under-estimated, in part due to the spatial dispersion of oocysts in the waterbody, but predominantly from imperfect detection methods. The development of 'event-driven' monitoring schemes with barrier performance-based treatment verification methods, as promoted in new guidelines, is supported as a pressing issue to reduce the likelihood of undetected pathogen passage through a treatment plant.

Animals↗

PubMed2005

Environmental inactivation of Cryptosporidium oocysts in catchment soils.

AIMS: To generate field-relevant inactivation rates for Cryptosporidium oocysts in soil that may serve as parameter values in models to predict the terrestrial fate and transport of oocysts in catchments. METHODS AND RESULTS: The inactivation of Cryptosporidium oocysts in closed soil microcosms over time was monitored using fluorescence in situ hybridization (FISH) as an estimate of oocyst 'viability'. Inactivation rates for Cryptosporidium in two soils were determined under a range of temperature, moisture and biotic status regimes. Temperature and soil type emerged as significantly influential factors (P < 0.05) for Cryptosporidium inactivation. In particular, temperatures as high as 35 degrees C may result in enhanced inactivation. CONCLUSIONS: When modelling the fate of Cryptosporidium oocysts in catchment soils, the use of inactivation rates that are appropriate for the specific catchment climate and soil types is essential. FISH was considered cost-effective and appropriate for determining oocyst inactivation rates in soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Previous models for predicting the fate of pathogens in catchments have either made nonvalidated assumptions regarding inactivation of Cryptosporidium in the terrestrial environment or have not considered it at all. Field-relevant inactivation data are presented, with significant implications for the management of catchments in warm temperate and tropical environments.

Animals↗

PubMed2005

Quantifying the impact of runoff events on microbiological contaminant concentrations entering surface drinking source waters.

Concentrations of microbiological contaminants in streams increase during rainfall-induced higher flow 'event' periods as compared to 'baseflow' conditions. If the stream feeds a drinking water reservoir, such periods of heightened pathogen loads may pose a challenge to the water treatment plant and subsequently a health concern to water consumers downstream. In order to manage this risk, it is desirable to first quantify the differences in surface water quality between baseflow and event conditions. The Event Mean Concentration (EMC) is a flow-weighted average concentration of a contaminant over the duration of a single event, proposed here as a standard parameter for quantifying the net effect of events on microbial water quality. Application of the EMC concept was assessed using flow and quality data for several events from an urbanised catchment. Expected mean EMCs were significantly larger than expected mean baseflow concentrations (p-value< or =0.012) for three microbial agents - Escherichia coil (13,000 [n = 7] v. 610 [n = 16] mpn/100 ml), Cryptosporidium (234 [n = 6] v. 51 [n = 16] oocysts/10 litres) and Campylobacter (48 [n = 5] v. 2.1 [n = 16] mpn/100ml). These parameter estimates were complemented by estimating data variability and uncertainty in the form of second-order random variables. As such the results are in a format appropriate for potential use as components in probabilistic risk assessments evaluating the effect runoff events have on drinking water quality.

Animals↗

PubMed2005

Effluent quality from 200 on-site sewage systems: design values for guidelines.

The quality of effluent from an on-site sewage treatment system is a critical factor in designing the disposal area and, hence, ensuring the sustained performance of the system. Contaminant concentrations in effluent are typically specified in regulatory guidelines or standards; however, the accuracy of these guideline values are brought into question due to the poor performance of septic tanks and the high failure rates of disposal systems reported here and elsewhere. Results from studies of septic tank effluent quality indicated that the effluent is of poorer quality than currently suggested by guidelines. Aerated wastewater treatment systems were found to perform to accreditation guidelines; however, insufficient nutrient data is presently available to assess nutrient loads. It is proposed that the 80th percentile of system performance be adopted as the design value for sizing effluent disposal areas to minimise failure associated with overloading. For septic tanks this equates to 660 mg L(-1) SS, 330 mg L(-1) BOD, 250 mg L(-1) TN and 36 mg L(-1) TP.

Guidelines as Topic↗

PubMed2005

Biofilms in an urban water distribution system: measurement of biofilm biomass, pathogens and pathogen persistence within the Greater Stockholm Area, Sweden.

Distribution pipe biofilms can provide sites for the concentration of a wide range of microbial pathogens, thereby acting as a potential source of continual microbial exposure and furthermore can affect the aesthetic quality of water. In a joint project between Stockholm Water, the MISTRA "Sustainable Urban Water" program, the Swedish Institute for Infectious Disease Control and the Royal Technical University, Stockholm, the aim of the current study was to investigate biofilms formed in an urban water distribution system, and quantify the impact of such biofilms on potential pathogen accumulation and persistence within the Greater Stockholm Area, Sweden. When used for primary disinfection, ultra-violet (UV) treatment had no measurable influence on biofilm formation within the distribution system when compared to conventional chlorination. Biofilms produced within a model pilot-plant were found to be representative to those that had formed within the larger municipal water distribution system, demonstrating the applicability of the novel pilot-plant for future studies. Polystyrene microspheres (1.0 microm) and Salmonella bacteriophages demonstrated their ability to accumulate and persist within the model pilot-plant system, where the means of primary disinfection (UV-treatment, chlorination) had no influence on such phenomena. With the exception of aeromonads, potential pathogens and faecal indicators could not be detected within biofilms from the Stockholm water distribution system. Results from this investigation may provide information for water treatment and distribution management strategies, and fill key data gaps that presently hinder the refinement of microbial risk models.

Aeromonadaceae↗

PubMed2005

A deterministic model to quantify pathogen loads in drinking water catchments: pathogen budget for the Wingecarribee.

This paper describes the development and testing of a mathematical model as a tool to quantify pathogen loads in Sydney's drinking water catchments. It has been used to identify, quantify and prioritise sources of Cryptosporidium, Giardia and E. coli in the Wingecarribee catchment. The pathogen model promotes understanding of the relative significance of different sources of pathogen risks as well as their fate and transport as they move through the subcatchments. This pathogen model not only enables water utility managers to identify those catchment segments that may contribute the highest load of pathogens, but also where management options will be most effective.

Animals↗

PubMed2005

Evidence for the existence of Cryptosporidium oocysts as single entities in surface runoff.

There is uncertainty whether Cryptosporidium oocysts attach to particles or to each other under ambient water conditions. Particle size distributions of Cryptosporidium oocyst suspensions were determined over a range of ionic strengths and pHs to determine under those environmental conditions that may promote oocyst aggregation. Cryptosporidium oocysts were shown to only aggregate in high ionic strength solutions (>0.45 M) and remain largely as single entities at ionic strengths and pHs that were likely to be encountered in surface runoff. Similarly, in loam soil suspensions, rather than attaching to the soil particles the majority of oocysts also remained as single entities. Overall, oocysts are expected to remain largely unattached to either themselves or soil particles in overland runoff. This has implications for pathogen transport and modelling since oocysts that are freely suspended are more likely to be transported in runoff to surface waters than if attached to more dense soil/faecal particles.

Animals↗

PubMed2004

Fluorescence resonance energy transfer (FRET)-based specific labeling of Cryptosporidium oocysts for detection in environmental samples.

BACKGROUND: Accurate detection and quantification of Cryptosporidium oocysts in water are a challenge to the water industry. This article demonstrates a way to fluorescently label Cryptosporidium oocysts, based on fluorescence resonance energy transfer (FRET). Labeled oocysts can then be applied to environmental waters and their movement followed by flow cytometric detection and enumeration of the FRET-labeled oocysts, as demonstrated here with environmental water samples. METHODS: Cryptosporidium oocysts were labeled with three fluorochromes, FITC, Texas red, and Cy7, that through FRET yielded a Stokes shift of approximately 272 nm with excitation from a standard argon laser emitting at 488 nm. Defined flow cytometric settings and gatings were used to select FITC/green (530-nm), Texas red/red (650-nm), and Cy7/infrared (780-nm) fluorescing particles with light scatter properties similar to oocysts. Water concentrates were seeded with 10 tri-labeled oocysts and were analyzed using flow cytometry. Unseeded water concentrates were also analyzed. RESULTS: Analysis of unseeded water concentrates detected no autofluorescent particle similar to the labeled oocysts. Labeled oocysts were detected successfully with up to 85% recovery in water concentrates spiked with 10 tri-labeled oocysts. CONCLUSIONS: Low numbers of FRET-labeled oocysts can be quantified and clearly distinguished from autofluorescing background in environmental water concentrates.

Animals↗

PubMed2004

The fate of legionellae within distribution pipe biofilms: measurement of their persistence, inactivation and detachment.

Distribution pipe biofilms present a currently unquantified public health risk to consumers receiving water for domestic potable and non-potable use. The aim of this study was to quantify the numbers of legionellae, used here as model bacterial pathogens, that may accumulate, persist within and detach from distribution pipe biofilms. L. pneumophila recovered by standard culture from an 8 week-old biofilm formed within a novel pilot-scale water distribution system represented 1% of those present in the adjacent bulk water. A combined chlorine concentration exceeding 0.2 mg x L(-1) eliminated culturable sessile legionellae altogether, though the reduction in FISH-positive cells represented just 75+/-25% of the original amount, compared to a 5-log reduction in culturable cells during the same period. Where there was < 0.1 mg x L(-1) combined chlorine, an exponential decay/loss of sessile L. pneumophila was observed (k = 0.37 - 0.41) over the course of a 38-day experimental period. The inoculation of the system with 1 microm fluorescent microspheres and legionellae demonstrated that removal of the latter was dominated by chemical disinfection, with erosion and biological grazing playing lesser roles. Under turbulent (Re approximately 5000) conditions, larger clusters of biofilm become detached from substrata, with more than 90% of sessile legionellae mobilised into the bulk water phase. Interaction with both biofilms and a thermophilic Acanthamoeba isolate reduced the susceptibility of legionellae to thermal inactivation by between one and two orders of magnitude, though it increased their sensitivity to chemical (free and combined chlorine) disinfection.

Biofilms↗

PubMed2004

Assessing the health implications of turbidity and suspended particles in protected catchments.

The supply of unfiltered disinfected drinking water from Melbourne's fully protected catchments means that the water-quality managers must ensure that the source water poses no public health risk. High turbidity is currently used as a surrogate of pathogens, and harvesting of water is based on its measurement. The work presented here summarises suspended particle and associated pathogen, microbial indicator and faecal biomarker concentrations collected to (a) quantify turbidity in an Australian water supply system and (b) assess the possibility of increasing water harvesting from selected tributaries. Pathogens and microbial indicators were present in low numbers in these source waters; increased turbidity during storm events was not associated with an increase in pathogen concentration. The results confirmed that protected catchments, along with good management, were effective barriers to pathogen contamination. Aesthetic issues still need to be addressed, but no measurable increase in microbiological risk was associated with storm-generated particles.

Bacteria↗

PubMed2004

QMRA (quantitative microbial risk assessment) and HACCP (hazard analysis and critical control points) for management of pathogens in wastewater and sewage sludge treatment and reuse.

Hazard Analysis and Critical Control Points (HACCP) was applied for identifying and controlling exposure to pathogenic microorganisms encountered during normal sludge and wastewater handling at a 12,500 m3/d treatment plant utilising tertiary wastewater treatment and mesophilic sludge digestion. The hazardous scenarios considered were human exposure during treatment, handling, soil application and crop consumption, and exposure via water at the wetland-area and recreational swimming. A quantitative microbial risk assessment (QMRA), including rotavirus, adenovirus, haemorrhagic E. coli, Salmonella, Giardia and Cryptosporidium, was performed in order to prioritise pathogen hazards for control purposes. Human exposures were treated as individual risks but also related to the endemic situation in the general population. The highest individual health risk from a single exposure was via aerosols for workers at the belt press for sludge dewatering (virus infection risk = 1). The largest impact on the community would arise if children ingested sludge at the unprotected storage site, although in the worst-case situation the largest number of infections would arise through vegetables fertilised with sludge and eaten raw (not allowed in Sweden). Acceptable risk for various hazardous scenarios, treatment and/or reuse strategies could be tested in the model.

Animals↗

PubMed2004

Detection, integration and persistence of aeromonads in water distribution pipe biofilms.

The occurrence of Aeromonas spp. within biofilms formed on stainless steel (SS), unplasticized polyvinyl chloride (uPVC) and glass (GL) substrata was investigated in modified Robbins Devices (MRD) in potable (MRD-p) and recycled (MRD-r) water systems, a Biofilm Reactor (BR) and a laboratory-scale pipe loop (PL) receiving simulated recycled wastewater. No aeromonads were isolated from the MRD-p whereas 3-10% of SS and uPVC coupons (mean 3.85 CFU cm(-2) and 12.8 CFU cm(-2), respectively) were aeromonad-positive in the MRD-r. Aeromonads were isolated from six SS coupons (67%) (mean 63.4 CFU cm(-2)) and nine uPVC coupons (100%) (mean 6.50x 10(2) CFU cm(-2)) in the BR fed with recycled water and from all coupons (100%) in the simulated recycled water system (PL). Mean numbers of aeromonads on GL and SS coupons were 5.83 x 10(2) CFU cm(-2) and 8.73 x 10(2) CFU cm(-2), respectively. No isolate was of known human health significance (i.e. Aeromonas caviae, A. hydrophila or A. veronii), though they were confirmed as Aeromonas spp. by PCR and fluorescence in situ hybridization (FISH). Challenging the PL biofilms with a slug dose of A. hydrophila (ATCC 14715) showed that biofilm in the PL accumulated in the order of 10(3)-10(4) A. hydrophila cm(-2), the number of which decreased over time, though could not be explained in terms of conventional 1st order decay kinetics. A sub-population of FISH-positive A. hydrophila became established within the biofilm, thereby demonstrating their ability to incorporate and persist in biofilms formed within distribution pipe systems. A similar observation was not made for culturable aeromonads, though the exact human health significance of this remains unknown. These findings, however, further question the adequacy of culture-based techniques and their often anomalous discrepancy with direct techniques for the enumeration of bacterial pathogens in environmental samples.

Aeromonas↗

PubMed2003

Particle dispersion for further Cryptosporidium and Giardia detection by flow cytometry.

AIMS: The aim of this study was to overcome the analytical problems encountered during the detection of protozoans by flow cytometry resulting from particle compaction. METHODS AND RESULTS: Malvern Mastersizer (Malvern Instruments, Malvern, UK) was used to characterize the particle distribution of four different water samples and/or particle concentrates incubated with (i) low ionic strength solution or sequestring agent, (ii) anionic or non-ionic surfactants (iii) industry detergent formulations and (iv) physical treatment. The recovery of oocysts and cysts in seeded and treated particle concentrates was estimated by cytometry and microscopy. The decrease in ionic strength of the aqueous solution was most efficient in particle dispersion for different types of water. Moreover, samples treated with deionized water or tetrasodium pyrophosphate showed the highest recovery with more than 80% of the oocysts and cysts recovered. CONCLUSIONS: Chemical treatments that act by altering the ionic strength of the medium are the most efficient for all water types tested here but the overall detergency performance cannot be predicted for all water types. SIGNIFICANCE AND IMPACT OF THE STUDY: Flow cytometric detection has been replaced largely by immunomagnetic separation but the data recorded still have relevance in this technique as well as in molecular techniques requiring DNA or RNA extraction.

Animals↗

PubMed2003

A theoretical approach to assess microbial risks due to failures in drinking water systems.

A failure in treatment or in the distribution network of a surface water-works could have serious consequences due to the variable raw water quality in combination with an extended distribution. The aim of this study was to examine the theoretical impact of incidents in the drinking water system on the annual risk of infection in a population served by a large water treatment plant in Sweden. Reported incidents in the system were examined and a microbial risk assessment that included three pathogens, Cryptosporidium parvum, rotavirus and Campylobacter jejuni, was performed. The main risk incidents in water treatment were associated with sub-optimal particle removal or disinfection malfunction. Incidents in the distribution network included cross-connections and microbial pollution of reservoirs and local networks. The majority of the annual infections were likely to be due to pathogens passing treatment during normal operation and not due to failures, thus adding to the endemic rate. Among the model organisms, rotavirus caused the largest number of infections. Decentralised water treatment with membranes was also considered in which failures upstream fine-pored membranes would have little impact as long as the membranes were kept intact.

Disinfection↗

PubMed2003

Enteric virions and microbial biofilms--a secondary source of public health concern?

Through their many sorption sites, microbial biofilms can accumulate both organic and inorganic particulate and colloidal material from bulk water environments. An application of such first principles to the ecology of "biocolloidal" enteric virions would suggest that they too may be concentrated by biofilms in a similar way. Though previous studies have isolated human gastrointestinal (enteric) virions from microbial biofilms, the exact human health significance of this has been neither fully investigated nor completely understood. Through an assessment of the location, accumulation and persistence of model enteric virion (phiX174, MS2 and B40-8 bacteriophages as well as 20 nm fluorescent latex microspheres) within biofilms, the aim of the current study was to investigate whether the interaction of enteric virions with distribution pipe biofilms could provide a secondary source of public health concern to consumers. Model enteric virions were found to be incorporated into biofilms at concentrations representing 1% of those present in the adjacent bulk water environment. A sub-population (0.01%) of these persisted throughout an experimental period of 30 days, inferring their potential to accumulate over time. Furthermore, model enteric virions were partitioned into bacterial microcolonies, environments where biofilm bacteria can persist and re-grow, even in the presence of "acceptable" levels of disinfection. A risk model for enteric virion accumulation and release from distribution pipe biofilms suggested that associated risks may exceed USEPA benchmark values. These findings could have wide-reaching implications in water treatment and distribution strategies, and necessitate a re-appraisal of current water guideline values.

Bacteria↗