A comparison of the role of two blue-green algae in THM and HAA formation

The contribution of two blue-green algae species, Anabaena flos-aquae and Microcystis aeruginosa, to the formation of trihalomethanes (THMs) and haloacetic acids (HAAs) was investigated. The experiments examined the formation potential of these disinfection by-products (DBPs) from both algae cells and extracellular organic matter (EOM) during four algal growth phases. Algal cells and EOM of Anabaena and Microcystis exhibited a high potential for DBP formation. Yields of total THMs (TTHM) and total HAAs (THAA) were closely related to the growth phase. Reactivity of EOM from Anabaena was slightly higher than corresponding cells, while the opposite result was found for Microcystis. Specific DBP yields (yield/unit C) of Anabaena were in the range of 2-11 μmol/mmol C for TTHM and 2-17 μmol/mmol C for THAA, while those of Microcystis were slightly higher. With regard to the distributions of individual THM and HAA compounds, differences were observed between the algae species and also between cells and EOM. The presence of bromide shifted the dominant compounds from HAAs to THMs.     

Comparison of potentially pathogenic free-living amoeba hosts by Legionella spp. in substrate-associated biofilms and floating biofilms from spring environments

This study compares five genera of free-living amoebae (FLA) hosts by Legionella spp. in the fixed and floating biofilm samples from spring environments. Detection rate of Legionella spp. was 26.9% for the floating biofilms and 3.1% for the fixed biofilms. Acanthamoeba spp., Hartmanella vermiformis, and Naegleria spp. were more frequently detected in floating biofilm than in fixed biofilm samples. The percentage of pathogenic Acanthamoeba spp. among all the genus Acanthamoeba detected positive samples was 19.6%. The potential pathogenic Naegleria spp. (for example, Naegleria australiensis, Naegleria philippinensis, and Naegleria italica) was 54.2% to all the Naegleria detected positive samples. In the study, 12 serotypes of possible pneumonia causing Legionella spp. were detected, and their percentage in all the Legionella containing samples was 42.4%. The FLA parasitized by Legionella included unnamed Acanthamoeba genotype, Acanthamoeba griffini, Acanthamoeba jacobsi, H. vermiformis, and N. australiensis. Significant differences were also observed between the presence/absence of H. vermiformis and Legionella parasitism in FLA. Comparisons between the culture-confirmed method and the PCR-based detection method for detecting FLA and Legionella in biofilms showed great variation. Therefore, using these analysis methods together to detect FLA and Legionella is recommended.     

Patterns of metal bioaccumulation in two filter-feeding macroinvertebrates: Exposure distribution, inter-species differences and variability across developmental stages

This study focused on the metal bioaccumulation of two aquatic insects (Ephoron virgo and Hydropsyche spp.) in order to evaluate the spatial distribution of metals, the interspecific differences between both filter-feeders and the bioaccumulation dynamics during E. virgo development stages. Hg, Cd, Ni, Cr, As, Pb, Cu, Ti, Zn and Mn were quantified in insects and in suspended particulate matter (SPM) sampled downstream and upstream of a chemical plant, where more than 300,000 t of polluted sediments are deposited. Hg concentrations were one order of magnitude higher downstream of the sediment dump, which showed that the Hg pollution originated in the chemical plant. Cd, Ni, Cr, Pb, Ti, Zn and Mn in invertebrates revealed that metal pollution was present upstream in other parts of the river. Interspecific differences were observed for all metals but Mn; significantly higher concentrations were observed in E. virgo over Hydropsyche exocellata, except for Cd, which showed 10-fold higher values. Hg and Cd increased until E. virgo nymphs reached 11 mm and decreased afterwards in late instars when nymphs were about to emerge. Cr, Pb, Ti and Mn decreased along early instars followed by a steady state in late instars. Similar values were obtained for Cu, As and Zn along all instars. Sexual differences between males and females of E. virgo were observed for Cd, Cu and Mn. Hg and Cd persistence was strong across developmental stages since high concentrations were found in eggs and emerging adults. Because the behavior of different metals varied for the two species and during the developmental stages of E. virgo, care should be taken in the interpretation of insect metal concentrations when analyzing the food chain transfer of metals in river ecosystems.     

Recommendations following a multi-laboratory comparison of microbial source tracking methods

Microbial source tracking (MST) methods were evaluated in the Source Identification Protocol Project (SIPP), in which 27 laboratories compared methods to identify host sources of fecal pollution from blinded water samples containing either one or two different fecal types collected from California. This paper details lessons learned from the SIPP study and makes recommendations to further advance the field of MST. Overall, results from the SIPP study demonstrated that methods are available that can correctly identify whether particular host sources including humans, cows and birds have contributed to contamination in a body of water. However, differences between laboratory protocols and data processing affected results and complicated interpretation of MST method performance in some cases. This was an issue particularly for samples that tested positive (non-zero C_t values) but below the limits of quantification or detection of a PCR assay. Although false positives were observed, such samples in the SIPP study often contained the fecal pollution source that was being targeted, i.e., the samples were true positives. Given these results, and the fact that MST often requires detection of targets present in low concentrations, we propose that such samples be reported and identified in a unique category to facilitate data analysis and method comparisons. Important data can be lost when such samples are simply reported as positive or negative. Actionable thresholds were not derived in the SIPP study due to limitations that included geographic scope, age of samples, and difficulties interpreting low concentrations of target in environmental samples. Nevertheless, the results of the study support the use of MST for water management, especially to prioritize impaired waters in need of remediation. Future integration of MST data into quantitative microbial risk assessments and other models could allow managers to more efficiently protect public health based on site conditions.     

Fluidized bed ash and passive treatment reduce the adverse effects of acid mine drainage on aquatic organisms

Elevated concentrations of acidity and metals in acid mine drainage (AMD) may be effectively addressed by active and passive treatment technologies. However, typical evaluations consider only chemical water quality with little if any regard for biological metrics. Robust evaluations including both chemical and biological indicators of water quality improvement are needed. In this study, injection of alkaline fluidized bed ash (FBA) into a flooded underground coal mine was coupled with a five-cell passive treatment system to ameliorate an abandoned AMD discharge in eastern Oklahoma. The passive system included process units promoting both aerobic and anaerobic treatment mechanisms. Resulting water quality changes and biological responses were evaluated. Organisms of two distinct functional groups (the filter-feeding mollusk Corbicula fluminea and the wide-spectrum feeding fish Lepomis macrochirus) were exposed to mine waters in several treatment cells. The combination of treatment technologies was hypothesized to limit potential negative effects on these aquatic organisms. Tissues were harvested and analyzed for concentrations of several metals (Al, Fe, Mn, Mg, Ca, Ni, Cu and Zn) of interest. Organismal responses, such as hepatosomatic index, condition factor, and condition index, did not vary significantly among organisms exposed within different treatment cells when compared to non-AMD impaired waters. Metal tissue accumulation trends, compared to aqueous concentrations, were observed for Fe, Ni and Zn. Exposure experiments with these two organisms indicated that FBA introductions coupled with passive treatment decreased the potential adverse effects of AMD to biological systems.     

Detection of Legionella spp. and some of their amoeba hosts in floating biofilms from anthropogenic and natural aquatic environments

Floating biofilms develop at the water-air interface and harbor numerous microorganisms, some of which are human pathogens like Legionella pneumophila. The presence of Legionella spp. and especially L. pneumophila in such biofilms was investigated. In parallel, the occurrence of Naegleria spp., Acanthamoeba spp., Willaertia spp., Vahlkampfia spp. and Hartmanella spp. was determined and it was examined whether Acanthamoeba spp. isolates were naturally infected with L. pneumophila bacteria. Eight anthropogenic and 37 natural aquatic environments were sampled between June and August 2005. Both Legionella spp. and L. pneumophila were present in 100% of the floating biofilms of the anthropogenic aquatic systems. Eighty-one percent of all natural floating biofilm samples were positive for Legionella spp. and 70% of these samples were positive for L. pneumophila. Legionella concentrations were in the range of 10(1)-10(2)cells/cm(2). Naegleria spp. and Acanthamoeba spp., two well-known L. pneumophila amoeba hosts, were present in 50-92% and 67-72% of floating biofilm samples, respectively. Acanthamoeba spp. isolates appeared to be naturally infected with L. pneumophila bacteria as proved by fluorescent in situ hybridization.     

The occurrence of campylobacters in water sources in South Africa

Campylobacter spp., mainly C. jejuni and C. coli, are recognized as significant human bacterial pathogens, being responsible for increasing numbers of gastroenteritis cases worldwide. Several reports have indicated that environmental waters are potential reservoirs and transmitting vehicles for these bacteria. The purpose of this study was thus to examine the occurrence of campylobacters in drinking and environmental water sources of South Africa, a country with a warmer climate and higher microbial pollution levels than those previously addressed in the Northern Hemisphere where similar investigations have been undertaken. Various types of water samples (five drinking water, four ground water, 11 surface water and four raw sewage) were collected from different parts of South Africa. Detection was by enrichment in Bolton broth prior to plating on both selective mCCDA or through a 0.6microm membrane filter on non-selective blood agar isolation media. Out of 100 initially selected Campylobacter-like isolates, only 22 did not grow aerobically and were subsequently identified as Campylobacter spp. by biochemical tests. However, the results obtained by 16S rRNA sequence analysis indicated that only three of these strains (13.6%) were Campylobacter jejuni and the remaining 19 strains were identified as Arcobacter butzleri. The spread of Arcobacter via water warrants further investigation, especially in view of the higher levels of detection and pathogenic nature of these bacteria.     

Monitoring the role of aceticlasts in anaerobic digestion: activity and capacity

Aceticlastic methanogens are seen as a key to digester capacity and stability. This paper develops and applies an assay to measure digester stability by measuring the maximum aceticlastic methane production rate (V_max,ac). The V_max,ac in combination with acetate concentrations was found to be an effective digestion monitoring tool to indicate process upsets. At steady state, thermophilic, first stage and short SRT digesters generally had a greater V_max,ac than mesophilic, second stage or long SRT digesters. The ratio of the V_max,ac to the plant aceticlastic methane production rate, termed the Acetate Capacity Number (ACN), is a measure of the excess capacity of the digester. Either V_max,ac or ACN can be used to estimate the capability to handle higher organic loading rates. Monod modeling was used to predict V_max,ac, ACN and maximum VS loading rates for mesophilic and thermophilic digestion and for staged digesters to better understand expected digestion capacity and stability.     

Cryptosporidium spp. and Giardia duodenalis in two areas of Galicia (NW Spain)

The aim of the present study was to investigate the environmental dispersal of Cryptosporidium spp. and Giardia duodenalis in two distinct areas (coastal and inland) in Galicia (NW Spain). Faecal samples were collected from healthy asymptomatic domestic (cows and sheep) and wild animals (deer and wild boars) in the selected areas. In each of the selected areas, samples of untreated water (influent) and of treated water (final effluent) were collected from each of the 12 drinking water treatments plants (DWTPs) and 12 wastewater treatment plants (WTPs) under study. Analysis of a single sample from each of the 635 (coastal) and 851 (inland) domestic and wild animals selected at random revealed that the prevalences of cryptosporidiosis and giardiosis in coastal area were 9.2% and 15.9% respectively, and in inland area, 13.7% and 26.7% respectively. In the coastal area, Cryptosporidium spp. oocysts were detected in influent and effluent samples from 2/12 (16.6%) DWTPs and 8/12 (66.6%) WTPs, while G. duodenalis cysts were detected in influent and effluent samples from 3/12 (25.0%) DWTPs and 12/12 (100%) WTPs. The concentrations were notably higher in WTPs; the mean parasite concentrations in the final treated effluent were 10 oocysts per litre and 137.8 cysts per litre for Cryptosporidium and Giardia, respectively. The mean concentration of G. duodenalis cysts per litre was significantly higher (P < 0.05) than the mean concentration of Cryptosporidium spp. oocysts per litre in both the influent and the effluent samples from all the treatment plants. In the coastal area, C. parvum, C. hominis and G. duodenalis assemblages A (I and II) and E were most repeatedly detected. In the inland area, C. parvum, C. andersoni and G. duodenalis assemblages A (I and II), B and E were most frequently identified.     

Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

The contribution of fecal pollution from dogs in urbanized areas can be significant and is an often underestimated problem. Microbial source tracking methods (MST) utilizing quantitative PCR of dog-associated gene sequences encoding 16S rRNA of Bacteroidales are a useful tool to estimate these contributions. However, data about the performance of available assays are scarce. The results of a multi-laboratory study testing two assays for the determination of dog-associated Bacteroidales (DogBact and BacCan-UCD) on 64 single and mixed fecal source samples created from pooled fecal samples collected in California are presented here. Standardization of qPCR data treatment lowered inter-laboratory variability of sensitivity and specificity results. Both assays exhibited 100% sensitivity. Normalization methods are presented that eliminated random and confirmed non-target responses. The combination of standardized qPCR data treatment, use of normalization via a non-target specific Bacteroidales assay (GenBac3), and application of threshold criteria improved the calculated specificity significantly for both assays. Such measures would reasonably improve MST data interpretation not only for canine-associated assays, but for all qPCR assays used in identifying and monitoring fecal pollution in the environment.     

Developmental toxicity, uptake and distribution of sodium chromate assayed by frog embryo teratogenesis assay-Xenopus(FETAX)

The embryotoxicity and teratogenicity of Cr(VI) on the survival and morphology of the anuran Xenopus laevis have been assessed by frog embryo teratogenesis assay-Xenopus (FETAX). The lethal median (LC₅₀) and teratogenic median (TC₅₀) concentration values of Cr(VI) were 890 µM and 260 µM, respectively. The calculated teratogenic index (TI) value was 3.42, suggesting that hexavalent chromium has a teratogenic potential. Malformations of embryos included lifting of the body, coiling of the tail and body oedema. Furthermore, the chromium salt caused significant growth retardation at 25 µM exposure concentrations. The use of radiolabelled ⁵¹Cr(VI) allowed the determination of the time course uptake of Cr in Xenopus exposed to concentrations ranging from 0.025 to 500 µM. The evaluation of its distribution into the body (head-abdomen-tail) was evaluated at different exposure times. Chromium is taken up at 24 h by Xenopus embryos for all concentrations tested. At 48 h post fertilization (stage of larva) the amount of Cr accumulated by the two-day-old larva ranged from 0.42 to 580 pg mg^− 1 wet weight at 0.025 and 500 µM respectively. These amounts were lower than those at 24 h (2.77 to 11016 pg mg^− 1 wet weight embryo) reaching values of the same order of magnitude at 120 h (five-days-old larva). Since at 48 h Xenopus development leads to a swimming embryo, the observed uptake at 24 h could be the result of the binding of Cr to jelly coat compounds surrounding the embryo body as confirmed by gel filtration experiments on ⁵¹Cr-jelly coat. The interaction of Cr with jelly coat is in agreement with the role of jelly coat in protecting the embryo against pathogen and chemical toxins to ensure fertilization. This work further supports the hypothesis that Cr contamination of surface waters could contribute to explain the reported worldwide depletion of frog population.     

Detection of enteric viruses, Giardia and Cryptosporidium in two different types of drinking water treatment facilities

In this study, two types of drinking water treatment facilities (two conventional drinking water treatment plants (DWTPs) and two compact units (Cus)) were compared referring to their production capacity. Water samples were collected from three main points: (a) different water treatment steps (b) washings of sand filters and (c) distribution system at different distances from the water treatment plants. Both viruses and protozoa were concentrated from each water sample by adsorption and accumulation on the same nitrocellulose membrane filters (0.45 microm pore size). Enteroviruses were detected by plaque infectivity assay in BGM cells and HAV, HEV and Norovirus were detected by RT-PCR. Giardia and Cryptosporidium were detected by conventional staining methods and PCR. The results revealed that enterovirus load at the intake ranged between 10-15 PFU/L for the two compact units and between 4.5 and 75 PFU/L for the two conventional DWTPs. The virus load in distribution system of the first type DWTPs at 1 km from the plant was the same as that of the intake. Viruses in the other type of treatment plants CUs at 1, 5 and 7 km, were much reduced. Investigation of raw water sediments of the two DWTPs showed enterovirus counts between 12 and 17.5 PFU/L. Virus count was reduced in sand of filters after washing. Giardia cysts were equally detected by microscopy and PCR in only intake samples of EL-Hawamdia CU (33.3%) and Meet Fares DWTP (50%). Cryptosporidium oocysts were equally detected by microscopy and PCR in intake samples of Abo EL-Nomros CU (100%), EL-Hawamdia CU (66.7%) and Fowa DWTP (50%). At Meet Fares DWTP three positive intake samples for Cryptosporidium were detected by PCR, compared with only two positive samples by microscopy. Giardia cysts and Cryptosporidium oocysts were detected in raw water sediment and sand of filters before washing. Only one sample from Meet Fares DWTP sand of filters after washing was positive for both Giardia and Cryptosporidium. It can be concluded that the poor microbial quality of the water may be due to improper operational skills and management of the various water treatment plants (especially at the two high capacity treatment plants).     

Analytical comparison of nine PCR primer sets designed to detect the presence of Escherichia coli/Shigella in water samples

The analytical performance of 9 different PCR primer sets designed to detect Escherichia coli and Shigella in water has been evaluated in terms of ubiquity, specificity, and analytical detection limit. Of the 9 PCR primer sets tested, only 3 of the 5 primer sets targeting uidA gene and the primer set targeting tuf gene amplified DNA from all E. coli strains tested. However, of those 4 primer sets, only the primer set targeting the tuf gene also amplified DNA from all Shigella strains tested. For the specificity, only the primer sets targeting the uidA gene were 100% specific although the primer sets targeting 16S rRNA, phoE, and tuf genes only amplified Escherichia fergusonii as non-specific target. Finally, the primer set targeting the 16S-ITS-23S gene region, was not specific as it amplified DNA from many other Enterobacteriaceae species. In summary, only the assay targeting the tuf gene detected all E. coli/Shigella strains tested in this study. However, if it becomes important to discriminate between E. coli and E. fergusonii, assays targeting the uidA gene would represent a good choice although none of them were totally ubiquitous to detect of the presence of Shigella strains.     

Hepatic levels of metal and metallothioneins in two commercial fish species of the Northern Iberian shelf

Metal levels (Cd, Pb, Hg, Cr, Fe, Cu, Zn) together with metallothioneins (MTs) were determined in the liver of two commercial fish species collected along the Northern Iberian coast. The four-spotted megrim (Lepidorhumbos boscii) and the pouting (Trisopterus luscus) were selected as representative species of the middle/outer (200-500 m) and inner shelf (70-120 m), respectively. Metal pollution in the middle/outer shelf was mostly detected in the Asturias and Basque Country areas, whereas in the inner shelf, pollution was widely spread along the coast. Significantly high levels of Pb, Cd, Cr and Hg were detected in T. luscus from different sampling sites and associated to anthropogenic pressure. MTs were induced in fish that had higher amounts of Zn, Cd, Cr and Hg in their livers, supporting the use of MT as a biomarker of metal exposure. The study supports the usefulness of the selected fish species as sentinel organisms for future biomonitoring studies.     

Kinetic characterisation of an enriched Nitrospira culture with comparison to Nitrobacter

Nitrospira and Nitrobacter are nitrite-oxidising bacteria commonly identified in nitrogen removal wastewater treatment plants. Little is known about the growth parameters of Nitrospira or the effects of environmental conditions or inhibitory compounds on Nitrospira activity. These bacterial properties were investigated using an enriched Nitrospira culture and an enriched Nitrobacter culture or Nitrobacter literature values. Compared to Nitrobacter, Nitrospira was found to have a comparable optimal pH range (8.0-8.3); similar normalised activity-temperature relationship (0.44e(0.055(T-15))) for temperatures between 15 and 30 degrees C and a similar oxygen half-saturation constant, K(O) (0.54+/-0.14 mgL(-1)). The major differences identified were that Nitrospira had a lower nitrite half-saturation constant, K(S) (0.9+/-0.07 mgNO(2)-NL(-1)); lower inhibition threshold concentrations for free ammonia (between 0.04 and 0.08 mg NH(3)-NL(-1)) and free nitrous acid (less than 0.03 mg HNO(2)-NL(-1)) and a higher yield (0.15+/-0.04 g VSS g N(-1)). Therefore, Nitrospira is more likely to dominate nitrite oxidation under conditions with low ammonium and nitrite concentrations, which would provide an advantage to them due to their lower K(S) value while avoiding any free ammonia or free nitrous acid inhibition.     

Optimization of DGGE community fingerprinting for characterizing Escherichia coli communities associated with fecal pollution

We evaluated the use of DGGE fingerprinting to differentiate communities of Escherichia coli from animal and geographic sources. An initial screening of 15 gene candidates revealed the ability of three target genes (mdh, phoE and uidA-4) to effectively differentiate E. coli communities originating in horses, pigs, geese and goats. Cluster and jackknife analyses performed on the communities from a more extensive number of hosts (n = 150) including humans (via raw sewage), horses, pigs, geese and cows revealed that the internal accuracy of classification of E. coli community fingerprints to their origin was similar for each of the three genes (85-86%). Each of the three genes were tested for their ability to associate E. coli source- and sink communities in two settings featuring contaminated water; (i) a stream receiving municipal wastewater effluent and (ii) a pond inhabited by geese. For each gene, DGGE fingerprints effectively matched effluent- and downstream E. coli communities (98-100% similarity) and excluded upstream communities, while communities from goose fecal material were 77-79% similar to communities in pond water, indicating fecal inputs from geese. Furthermore, each gene discriminated against E. coli communities from hosts non-indigenous to either setting. DGGE analysis of E. coli communities appears to be a promising tool to augment existing efforts aiming to address the dynamics of bacteria pollution in complex, natural environments.     

Occurrence of Mycobacterium avium subsp. paratuberculosis in raw water and water treatment operations for the production of potable water

Mycobacterium avium subsp. paratuberculosis (Map) causes Johne’s disease of cattle and is implicated as a cause of Crohn’s disease in humans. The organism is excreted in animal faeces and can contaminate water catchment areas. This coupled with Map’s survival in the environment means that water destined for domestic use may be a source of exposure. This work was designed to determine the occurrence of Map in Lough Neagh (the largest freshwater lake in the British Isles), used as a reservoir, and in two water treatment works (WTW1 and WTW2) which abstract from the lough and which have slow sand filtration (SSF) and dissolved air flotation respectively as their principal treatment regimes. The organism was not detected in lough water samples by culture (n = 70) but 29% (20/70) were positive by PCR. In the raw water to WTW1 and WTW2 no culture positives were detected but 54% (13/24) and 58% (14/24) respectively were PCR positive. In WTW1 there were no culture positives at the SSF or final water but 31% (8/26) and 45% (9/20) respectively were PCR positive. In WTW2 similar results were obtained with 26% (6/23) and 48% (11/23) in the floccules and final water respectively. At WTW2 however one culture positive was detected in the final water. This latter finding is of concern. The inability to reach definitive conclusions indicates the need for further research, particularly in the detection methods for viable Map.     

Comparison of Enterococcus measurements in freshwater at two recreational beaches by quantitative polymerase chain reaction and membrane filter culture analysis

Cell densities of the fecal pollution indicator genus, Enterococcus, were determined by a rapid (3 h or less) quantitative polymerase chain reaction (QPCR) analysis method in 100 ml water samples collected from recreational beaches on Lake Michigan and Lake Erie during the summer of 2003. Measurements by this method were compared with counts of Enterococcus colony-forming units (CFU) determined by Method 1600 membrane filter (MF) analysis using mEI agar. The QPCR method had an estimated 95% confidence, minimum detection limit of 27 Enterococcus cells per sample in analyses of undiluted DNA extracts and quantitative analyses of multiple lake water samples, spiked with known numbers of these organisms, gave geometric mean results that were highly consistent with the spike levels. At both beaches, the geometric means of ambient Enterococcus concentrations in water samples, determined from multiple collection points during each sampling visit, showed approximately lognormal distributions over the study period using both QPCR and MF analyses. These geometric means ranged from 10 to 8548 cells by QPCR analysis and 1-2499 CFU by MF culture analysis in Lake Michigan (N=56) and from 8 to 8695 cells by QPCR and 3-1941 CFU by MF culture in Lake Erie (N=47). Regression analysis of these results showed a significant positive correlation between the two methods with an overall correlation coefficient (r) of 0.68.     

Quantitative risk assessment of Cryptosporidium in tap water in Ireland

Cryptosporidium species are protozoan parasites associated with gastro-intestinal illness. Following a number of high profile outbreaks worldwide, it has emerged as a parasite of major public health concern. A quantitative Monte Carlo simulation model was developed to evaluate the annual risk of infection from Cryptosporidium in tap water in Ireland. The assessment considers the potential initial contamination levels in raw water, oocyst removal and decontamination events following various process stages, including coagulation/flocculation, sedimentation, filtration and disinfection. A number of scenarios were analysed to represent potential risks from public water supplies, group water schemes and private wells. Where surface water is used additional physical and chemical water treatment is important in terms of reducing the risk to consumers. The simulated annual risk of illness for immunocompetent individuals was below 1 × 10^− 4 per year (as set by the US EPA) except under extreme contamination events. The risk for immunocompromised individuals was 2-3 orders of magnitude greater for the scenarios analysed. The model indicates a reduced risk of infection from tap water that has undergone microfiltration, as this treatment is more robust in the event of high contamination loads. The sensitivity analysis highlighted the importance of watershed protection and the importance of adequate coagulation/flocculation in conventional treatment. The frequency of failure of the treatment process is the most important parameter influencing human risk in conventional treatment. The model developed in this study may be useful for local authorities, government agencies and other stakeholders to evaluate the likely risk of infection given some basic input data on source water and treatment processes used.     

In vitro estimates of bioaccessible nickel in field-contaminated soils, and comparison with in vivo measurement of bioavailability and identification of mineralogy

This study determined nickel (Ni) bioaccessibility in weathered smelter-contaminated soils, separately for particle-sized fractions using two in vitro methods: simulated gastrointestinal digestion (PBET) and PBET followed by absorption by Caco-2 cells. Relative bioavailability of Ni in soils was determined in vivo using rats, validating in vitro estimates; a mineralogical basis of variation in bioavailability/bioaccessibility among soils was explored. In vitro assays identified the same difference in bioaccessibility for Ni among particle size fractions. PBET estimates were more precise, thus likely to be more useful in discerning differences among soils. In vivo bioavailability for Ni was below limit of detection for the small soil particles, and 31% and 56% for the larger particles. The relative bioavailability calculated from this work suggests that risk from ingesting Ni-contaminated soils could be overestimated by between 2- and 50-fold if the estimates of exposure are not adjusted for the lower bioavailability of weathered Ni originating from smelter emissions. The overestimation that would occur by using total Ni is greatest for the particle size that is most likely to adhere to the hands of children, demonstrating the importance of particle-size separation of soils for bioavailability determination and risk assessment.