Predicting bioremediation of hydrocarbons: laboratory to field scale.

There are strong drivers to increasingly adopt bioremediation as an effective technique for risk reduction of hydrocarbon impacted soils. Researchers often rely solely on chemical data to assess bioremediation efficiently, without making use of the numerous biological techniques for assessing microbial performance. Where used, laboratory experiments must be effectively extrapolated to the field scale. The aim of this research was to test laboratory derived data and move to the field scale. In this research, the remediation of over thirty hydrocarbon sites was studied in the laboratory using a range of analytical techniques. At elevated concentrations, the rate of degradation was best described by respiration and the total hydrocarbon concentration in soil. The number of bacterial degraders and heterotrophs as well as quantification of the bioavailable fraction allowed an estimation of how bioremediation would progress. The response of microbial biosensors proved a useful predictor of bioremediation in the absence of other microbial data. Field-scale trials on average took three times as long to reach the same endpoint as the laboratory trial. It is essential that practitioners justify the nature and frequency of sampling when managing remediation projects and estimations can be made using laboratory derived data. The value of bioremediation will be realised when those that practice the technology can offer transparent lines of evidence to explain their decisions.

Effects of copper and the sea lice treatment Slice on nutrient release from marine sediments.

Copper-based antifoulant paints and the sea lice treatment Slice are widely used, and often detectable in the sediments beneath farms where they are administered. Ten-day, whole sediment mesocosm experiments were conducted to examine how increasing sediment concentrations of copper or Slice influenced final water column concentrations of ammonium-nitrogen (NH(4)-N), nitrate+nitrite-nitrogen (NO(X)-N) and phosphate-phosphorus (PO(4)-P) in the presence of the non-target, benthic organisms Corophium volutator and Hediste diversicolor. Nominal sediment concentrations of copper and Slice had significant effects on the resulting concentrations of almost all nutrients examined. The overall trends in nutrient concentrations at the end of the 10-day incubations were highly similar between the trials with either copper or Slice, irrespective of the invertebrate species present. This suggests that nutrient exchange from the experimental sediments was primarily influenced by the direct effect of copper/Slice dose on the sediment microbial community, rather than the indirect effect of reduced bioturbation/irrigation due to increased macrofaunal mortality.

Leaching of bioluminescent Escherichia coli O157:H7 from sheep and cattle faeces during simulated rainstorm events.

AIMS: Development of a novel inoculation technique to improve the current methods of determining the leaching of Escherichia coli O157:H7 from faeces. METHODS AND RESULTS: Ruminant faeces were inoculated with a high [c. 10(7) colony forming units (CFU) g(-1)] or low (c. 10(4) CFU g(-1)) load of a lux-marked strain of E. coli O157:H7 via injection, and subjected to four simulated heavy rainfall events. The population density and metabolic activity of E. coli O157:H7 recovered within the leachate was determined following each simulated rain event and compared with the indigenous E. coli population. The concentration of E. coli O157:H7 in the leachates followed a similar trend to that of nonpathogenic E. coli. Significantly greater densities of generic and pathogenic E. coli were recovered in the leachates generated from sheep faeces compared with cattle faeces. Pathogen metabolic activity was also significantly greater in sheep faeces. CONCLUSIONS: Our findings show that E. coli O157:H7 may readily leach from ruminant faeces during rain events. The bacterium leaches more freely from sheep faeces than from cattle faeces and displays greater metabolic activity within sheep leachate. SIGNIFICANCE AND IMPACT OF THE STUDY: A novel inoculation technique was developed that allowed the determination of both population density and cellular activity of E. coli O157:H7 in leachate derived from faeces.

Survival of Escherichia coli O157:H7 in waters from lakes, rivers, puddles and animal-drinking troughs.

Survival of Escherichia coli O157:H7 in surface waters may increase the potential for dissipation of the organism to facilitate cycles of livestock re-infection and lead to human infection. Although previous studies have monitored survival of the organism in a range of surface waters, there is limited information on the influence of physico-chemical characteristics on persistence. Microcosms of four different surface water types (n=31) from the UK were inoculated with E. coli O157:H7 and incubated at 10 degrees C. The water types studied were: lake, puddle, river, and animal-drinking trough waters. Numbers of E. coli O157:H7 declined over time in all waters, although cells were still detected in 45% of non-sterile samples after 2 months. Persistence of E. coli O157:H7 was enhanced by water aeration and by prior sterilisation; however there was no correlation between water chemistry and mean E. coli O157:H7 die-off times or rates in any water type. Survival of the pathogen was better in lake and puddle waters than in river or drinking trough waters. Further studies are needed to establish the key water quality factors that regulate pathogen survival.

Physiological and toxicological characterization of an engineered whole-cell biosensor.

Bioluminescence-based bacterial biosensors are often reported as reliable and efficient tools for risk assessment and environmental monitoring. However, there are few data comparing the metabolism of genetically engineered strains to the corresponding wild type. A pollutant-degrading bacterium capable of mineralising 2,4-dichlorophenoxyacetic acid (2,4-D), Burkholderia sp. strain RASC c2, was genetically engineered to produce light constitutively and tested for assessing the main causes of biodegradation constraint affected by growth rates, toxicity, bioavailability and metal speciation in complex environments. This research focuses on such aspects by characterizing two pollutant-degrading isolates, the wild type and the genetically engineered biosensor (lux-marked). Degradation and growth rates of both isolates were assessed with different concentrations of 2,4-D as the sole carbon source. Kinetic rates were affected by initial concentration of substrate and isolates showed distinct growth rates at different 2,4-D concentrations. Toxic effects of zinc and copper were also comparatively assessed using a dehydrogenase assay and light output. The isolates were sensitive to both metals and at similar EC(50) values. Therefore, bioluminescence response of the lux-marked isolate accurately reflected the toxic response of the parental organism towards zinc and copper, making it an ideal test-organism for assessing toxicity in the context of pollutant mineralization.

Rhizoremediation of pentachlorophenol by Sphingobium chlorophenolicum ATCC 39723.

Sphingobium chlorophenolicum is well known as a pentachlorophenol (PCP) degrader. The objective of this study was to evaluate PCP degradation in a loamy sandy soil artificially contaminated with PCP using phytoremediation and bioaugmentation. Measurements of PCP concentrations were carried out using high performance liquid chromatography analyses (HPLC). The toxic effect of PCP on plants was studied through the monitoring of weight plant and root length. The biodegradation of PCP by S. chlorophenolicum in soil was assessed with a bioluminescence assay of Escherichia coli HB101 pUCD607. Bacterial analyses were carried out by plating on Mineral Salt Medium (MSM) for S. chlorophenolicum, MSM for PCP-degrading/tolerant organisms and Trypticase Soy Broth Agar (TSBA) for heterotrophic organisms. The introduction of S. chlorophenolicum into soil with plants showed a faster degradation when compared to the non-inoculated soil. The monitoring of the plant growth showed a protective role of S. chlorophenolicum against the toxicity of PCP. The bioassay confirmed that initial toxicity was lowered while degradation progressed. There was a significant increase of organisms tested in the roots in comparison to those in the soil. This study showed that the presence of S. chlorophenolicum enhanced the PCP degradation in a loamy soil and also it had a protective role to prevent phytotoxic effects of PCP on plant growth. The combined use of bioaugmentation and plants suggests that the rhizosphere of certain plant species may be important for facilitating microbial degradation of pesticides in soil with important implications for using vegetation to stabilize and remediate surface soils.

Salivary nitrate--an ecological factor in reducing oral acidity.

Human oral cavities represent a novel environment with a constant supply of concentrated nitrate. For humans, over 80% of dietary nitrate originates from fruit and vegetables. With a healthy, balanced diet, rich in fruit and vegetables, the concentration of nitrate in saliva can reach up to more than three times the European drinking water standard. The physiological function of the active excretion of salivary nitrate is unknown. Furthermore, little is known of the ecological function of oral nitrate and the effect on the oral environment during its subsequent oral microbial conversions. The objectives of the research were to investigate the effect on salivary pH coupled with oral microbial nitrate and/or nitrite reduction. Human saliva samples were incubated anaerobically in the presence of 111.0 mmol glucose (2%), with and without 1.5 mmol nitrate/nitrite, and pH and nitrate/nitrite consumption were measured during the time-course of the incubations. We found that anaerobic incubation of saliva containing a mixture of oral bacteria in the presence of nitrate/nitrite substrates and glucose resulted in a higher pH than was found in controls in the absence of nitrate/nitrite. These results suggest that the presence of these electron acceptors repressed acid fermentation, or increased alkali production, or consumed acid produced, thus reducing salivary acidity. This finding identifies salivary nitrate as a possible ecological factor in reducing oral acidity. The possibility that a symbiotic relationship between host nitrate excretion and nitrate-reducing microorganisms might help to protect against tooth decay should be explored further.

Survival of Escherichia coli O157:H7 in the rhizosphere of maize grown in waste-amended soil.

AIMS: To assess whether the persistence of Escherichia coli O157:H7 in soil amended with cattle slurry and ovine stomach content waste is affected by the presence of a maize rhizosphere. METHODS AND RESULTS: Cattle slurry and ovine stomach content waste were inoculated with E. coli O157:H7. Wastes were then applied to soil cores with and without established maize plants. The pathogen survived in soil for over 5 weeks, although at significantly greater numbers in soil receiving stomach content waste in comparison to cattle slurry. Persistence of the pathogen in soil was unaffected by the presence of a rhizosphere. CONCLUSIONS: Other factors may be more influential in regulating E. coli O157:H7 persistence in waste-amended soil than the presence or absence of a rhizosphere; however, waste type did have significant affect on the survival of E. coli O157:H7 in such soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Escherichia coli O157:H7 can be present within animal-derived organic wastes that are routinely spread on land. Introduced measures with regards to such waste disposal may decrease exposure to the organism; however, the persistence of E. coli O157:H7 for considerable periods in waste-amended soil may still pose some risk for both human and animal infection. This study has shown that whilst survival of E. coli O157:H7 in waste-amended soil is not significantly affected by the presence or absence of a maize rhizosphere; it may vary significantly with waste type. This may have implications for land and waste management.

Comparative evaluation of a bioluminescent bacterial assay in terrestrial ecotoxicity testing.

Despite the widespread and successful use of luminescence-based bioassays in water testing, their applications to soils and sediments is less proven. In part this is because such bioassays have mainly been carried out in an aqueous-based medium and, as such, favour contaminants that are readily water-soluble. In this study, aqueous solutions and soils contaminated with heavy metals (HM), polar organic contaminants and hydrophobic organic contaminants (HOCs) were tested using a range of luminescence-based bioassays (Vibrio fischeri, Escherichia coli HB101 pUCD607 and Pseudomonas fluorescens 10586r pUCD607). For the first two chemical groups, the assays were highly reproducible when optimised extraction procedures were employed but for HOCs the bioassay response was poor. Quantitative structure-activity relationships (QSARs) obtained from aqueous solutions had a linear response although correlation for the chemicals tested using bacterial bioassays was significantly less sensitive than that of sublethal tests for Tetrahymena pyriformis. Bacterial and Dendrobaena veneta bioassay responses to extracts from HM amended soils showed that a clear relationship between trophic levels could be obtained. There is no doubt that the wide range of bioluminescent-based bioassays offers complementary applications to traditional testing techniques but there is a significant need to justify and optimise the extraction protocol prior to application.

Influence of nematodes on resource utilization by bacteria--an in vitro study.

The positive influence of bacterial feeding nematodes on bacterial mediated processes such as organic matter mineralization and nutrient cycling is widely accepted, but the mechanisms of these interactions are not always apparent. Both transport of bacteria by nematodes, and nutritional effects caused by nematode N excretion are thought to be involved, but their relative importance is not known because of the difficulties in studying these interactions in soil. We developed a simple in vitro assay to study complex nematode/bacterial interactions and used it to conduct a series of experiments to determine the potential influence of nematode movement and nutritional effects on bacterial resource use. The system used bacterial feeding and nonfeeding insect parasitic nematodes, and luminescent bacteria marked with metabolic reporter genes. Both nutritional enhancement of bacterial activity and bacterial transport were observed and we hypothesize that in nature, the relative importance of transport is likely to be greater in bulk soil, whereas nematode excretion may have greater impact in the rhizosphere. In both cases, the ability of nematodes to enhance bacterial resource utilization has implications for soil components of biogeochemical cycling.

2,4-Dichlorophenoxyacetic acid (2,4-D) biodegradation in river sediments of Northeast-Scotland and its effect on the microbial communities (PLFA and DGGE).

A bench-scale study was conducted to investigate 2,4-D biodegradation rates at different concentrations (10, 100 and 1000 microg per gram of dry weight) in distinct sediments samples collected on the River Ythan, Northeast-Scotland. Mineralisation of 14C 2,4-D occurred mostly within 30 days for all tested concentrations with a degradation rate ranging from 5 to 750 microg d(-1). Biodegradation rates were affected by the biological and biochemical characteristics of the indigenous microbial community in the studied sediments rather than factors such as compound bioavailability and/or toxicity. PLFA-profiling provided evidences of the effect of 2,4-D amendments on the microbial communities and DGGE-profiling showed changes in the genetic potential of the microbial populations which might affect metabolic characteristics of the sediment. PLFAs biomarkers suggested that the pathway of alpha-ketoglutarate-dependent dioxygenase was the main route of 2,4-D biodegradation. This pathway is commonly found in microorganisms of the beta-subdivision of proteobacteria.

Persistence of Escherichia coli O157 on farm surfaces under different environmental conditions.

AIMS: To compare the persistence of Escherichia coli O157 on a variety of common faecally contaminated farmyard material surfaces (wood and steel) under different moisture and temperature regimes. METHODS AND RESULTS: Samples of field-conditioned farmyard materials (galvanized steel and wood) were cut into pieces and contaminated with fresh cattle faeces inoculated with nontoxigenic E. coli O157 (strain 3704). Thereafter, they were stored at four different environmental conditions; with temperature (5 and 20 degrees C) and moisture (moist or dry) as variables. Transfer of the pathogen to hands from the surfaces was also evaluated. Escherichia coli O157 numbers declined over time on all surfaces albeit at different rates according to the sample material and environmental conditions. Persistence was greatest on moist wood samples under cooler temperatures with large population numbers remaining after 28 days. Desiccation of surfaces resulted in a more rapid decline in E. coli O157 populations under both temperature regimes. Substantial numbers of colonies may also potentially be transferred to human hands from the surfaces during brief contact. CONCLUSIONS: When environmental conditions are favourable, E. coli O157 may persist for considerable times on a range of surfaces. However, when exposed to higher temperatures and dehydration, survival is notably decreased. Overall, bacterial persistence was significantly greater on wood samples relative to steel. SIGNIFICANCE AND IMPACT OF THE STUDY: Escherichia coli O157 is a prevalent pathogen, common in ruminant faeces. Contact with contaminated faeces may lead to human infection, resulting in possible severe illness. Although our study used only one strain of bacteria, our findings indicates that E. coli O157 has the potential to persist for long periods of time on gates, stiles and other farmyard surfaces under a range of environmental conditions. These farmyard surfaces therefore pose a potential infection pathway particularly where there is a high risk of direct human contact (e.g. child petting zoos, open farms).

Survival of E. coli O157:H7 in organic wastes destined for land application.

AIM: To determine the persistence of Escherichia coli O157 in contrasting organic wastes spread to land and to assess the potential environmental risk associated with the disposal of these wastes to land. METHODS AND RESULTS: Twenty-seven organic wastes originating from slaughterhouses, wastewater treatment plants (raw and treated sewage), creameries and farms (bovine slurry), were inoculated with E. coli O157:H7 and incubated at 10 degrees C. Although pathogen numbers gradually declined in all the wastes, albeit at different rates even in the same waste type, E. coli O157:H7 was still viable in 77% of organic wastes tested after 2 months. CONCLUSIONS: Long-term storage of organic wastes led to a significant and gradual decline in E. coli O157:H7 numbers. Consequently, storage may be a useful means of reducing the pathogen load of wastes destined for land application. However, in most cases, long-term storage cannot be expected to completely eliminate E. coli O157:H7 from waste. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results indicate that current legislation may be insufficient to protect the environment from E. coli O157:H7 contamination from untreated wastes spread to land.

Bacterial diversity promotes community stability and functional resilience after perturbation.

The relationships between bacterial community diversity and stability were investigated by perturbing soils, with naturally differing levels of diversity, to equivalent toxicity using copper sulfate and benzene. Benzene amendment led to large decreases in total bacterial numbers and biomass in both soils. Benzene amendment of an organo-mineral/improved pasture soil altered total soil bacterial community structure but, unlike amendment of the mineral/arable soil, maintained genetic diversity, based on polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis targeting DNA and RNA, until week 9 of the perturbation experiment. Assuming equivalent toxicity, the genetic diversity of the naturally more diverse soil was more resistant to benzene perturbation than the less diverse soil. The broad scale function (mineralization of 14C-labelled wheat shoot) of both benzene- and copper-treated soil communities was unaffected. However, narrow niche function (mineralization of 14C-labelled 2,4-dichlorophenol) was impaired for both benzene-polluted soils. The organo-mineral soil recovered this function by the end of the experiment but the mineral soil did not, suggesting greater resilience in the more diverse soil. Despite a large reduction in bacterial numbers and biomass in the copper-treated soils, only small differences in bacterial community diversity were observed by week 9 in the copper-polluted soils. The overall community structure was little altered and functionality, measured by mineralization rates, remained unchanged. This suggested a non-selective pressure and a degree of genetic and functional resistance to copper perturbation, despite a significant reduction in bacterial numbers and biomass. However, initial shifts in physiological profiles of both copper-polluted soils were observed but rapidly returned to those of the controls. This apparent functional recovery, accompanied by an increase in culturability, possibly reflects adaptation by the surviving communities to perturbation. The findings indicate that, although soil communities may be robust, relationships between diversity and stability need to be considered in developing a predictive understanding of response to environmental perturbations.

Root border cells take up and release glucose-C.

BACKGROUND AND AIMS: Border cells are released from the root tips of many plant species, and can remain viable in the rhizosphere for 1 week. Whether border cells are capable of controlled glucose exchange with their environment was investigated. METHODS: Border cells were removed from Zea mays L. root tips, and immersed in (14)C-labelled D-glucose. In one experiment, the hexose transport inhibitor, phlorizin, was used to investigate active glucose uptake from a range of glucose concentrations. In another experiment, glucose efflux from border cells was monitored over time. KEY RESULTS: Glucose uptake by the border cells increased with increasing glucose concentration from 0.2 to 20 mm. At 0.2 mm glucose, uptake was mainly active, as evidenced by the approx. 60 % inhibition with phlorizin. At 2 and 20 mm glucose, however, uptake was mainly via diffusion, as phlorizin inhibition was negligible. Glucose efflux increased with time for live border cells in both 2 and 20 mm glucose. There was no clear efflux/time pattern for heat-killed border cells. CONCLUSIONS: Border cells actively take up glucose, and also release it. Under our experimental conditions, glucose uptake and efflux were of similar order of magnitude. In the rhizosphere net glucose exchange will almost certainly depend on local soil conditions.

Influence of Soil Temperature and Matric Potential on Sugar Beet Seedling Colonization and Suppression of Pythium Damping-Off by the Antagonistic Bacteria Pseudomonas fluorescens and Bacillus subtilis.

ABSTRACT Pseudomonas fluorescens B5 and Bacillus subtilis MBI 600 colonized sugar beet seedlings at matric potentials of -7 x 10(3), -140 x 10(3), and -330 x 10(3) Pa and under five temperature regimes ranging from 7 to 35 degrees C, with diurnal fluctuations of 5 to 22 degrees C. No interaction between matric potential and temperature was observed. In situ bioluminescence indicated physiological activity of Pseudomonas fluorescens B5. Colonization of the root at >/=4 cm below the seed decreased at very low matric potential (-330 x 10(3) Pa). Total population size of Pseudomonas fluorescens B5 per seedling was significantly increased at -140 x 10(3) Pa. However, matric potential had no significant effect on the population density of Pseudomonas fluorescens per gram of root fresh weight and did not affect the distribution of the population down the root. Total population size per seedling and downward colonization by Pseudomonas fluorescens B5 were significantly reduced at high temperatures (25 to 35 degrees C). Maximum colonization down the root occurred at intermediate temperature (15 degrees C) at both matric potentials (-7 x 10(3) and -140 x 10(3) Pa). Addition of B. subtilis MBI 600 to the seed had no effect on rhizosphere populations of Pseudomonas fluorescens B5. Populations of B. subtilis MBI 600, which consisted largely of spores, were slightly reduced at lower matric potentials and were not affected by temperature. Survival and dry weight of plants in soils infested with Pythium spp. decreased with increasing soil temperature and matric potential, indicating an increase in disease pressure. However, there was no significant interaction between the two factors. At -330 x 10(3) Pa, soil dryness but not Pythium infection was the limiting factor for plant emergence. At temperatures of 7 to 25 degrees C and matric potentials of -7 x 10(3) to 120 x 10(3) Pa, treatment with Pseudomonas fluorescens B5 increased plant survival and dry weight. At 7 degrees C and -120 x 10(3) Pa, there was almost complete emergence of seeds treated with Pseudomonas fluorescens B5. Antagonistic activity of Pseudomonas fluorescens B5 decreased with increasing soil temperature and decreasing matric potential. At 25 to 35 degrees C and -7 x 10(3) Pa, no effect was observed. In regimes with different day and night temperatures, the maximum (day) temperature was decisive for disease development and antagonistic activity. B. subtilis MBI 600 displayed no significant antagonistic effect against Pythium ultimum and did not influence the performance of Pseudomonas fluorescens B5 in combined inocula.

Nematode-enhanced microbial colonization of the wheat rhizosphere.

The mechanisms by which seed-applied bacteria colonize the rhizosphere in the absence of percolating water are poorly understood. Without mass flow, transport of bacteria by growing roots or soil animals, particularly nematodes may be important. We used a sand-based microcosm system to investigate the ability of three species of nematodes (Caenorhabditis elegans, Acrobeloides thornei and a Cruznema sp.) to promote rhizosphere colonization by four strains of beneficial rhizobacteria. In nearly all cases, rhizosphere colonization was substantially increased by the presence of nematodes, irrespective of bacterial or nematode species. Our results suggest that nematodes are important vectors for bacteria rhizosphere colonization in the absence of percolating water.

Big bacteria pass through very small holes.

Can normal-sized bacteria pass through small holes, and if so how? We have found that common, potentially pathogenic, bacteria (which are nominally larger than 0.2 microm) can cross a 0.2 microm nylon membrane. All of the bacteria crossed from the upper membrane surface to the solid medium below the membrane; this ability was highly repeatable and did not depend on the make of membrane used. Bacteria growing below the membrane exhibited normal size and morphology. We suggest that the ability of common bacteria to pass through small holes has important implications in relation to their role as pathogens.

On-line microbial biosensing and fingerprinting of water pollutants.

The potential for biosensors to contribute to on-line toxicity testing for monitoring of water quality is currently constrained both by the relevance of the biosensors available and the technology for biosensor delivery. This paper reports the use of novel slow release biosensor delivery for on-line monitoring instrumentation, with environmentally relevant bacteria for both simple toxicity testing and more complex toxicity fingerprinting of industrial effluents. The on-line toxicity test, using bioluminescence-based biosensors, proved to be as sensitive and reliable as the corresponding batch test, with comparable contaminant EC(50) values from both methods. Toxicity fingerprinting through the investigation of the kinetics (dose-response) and the dynamics (response with time) of the biosensor test response proved to be diagnostic of both effluent type and composition. Furthermore, the slow release of biosensors immobilised in a polyvinyl alcohol (PVA) matrix greatly improved biosensor delivery, did not affect the sensitivity of toxicity testing, and demonstrated great potential for inclusion in on-line monitoring instrumentation.

A novel toxicity fingerprinting method for pollutant identification with lux-marked biosensors.

A novel technique is described for the identification and quantification of environmental pollutants based on toxicity fingerprinting with a metabolic lux-marked bacterial biosensor. This method involved characterizing the toxicity-based responses of the biosensor to seven calibration pollutants as acute temporal-dose response fingerprints. An algorithm is described to allow comparisons of responses of an unknown pollutant to be made against the calibration data. This is based on predicting pollutant concentration at each of six different time points over the course of a 5-min assay. If the prediction is consistent between the unknown pollutant and a calibration pollutant at the 95% test level, this is considered to be a positive identification. All seven calibration pollutants could be successfully distinguished from each other with this technique. Environmental samples, individually spiked with single concentrations of pollutants, were compared in this way against the calibration pollutants. An 83% identification success was achieved, with no false positives at the 95% test level. This is a simple and rapid technique that potentially can be applied to monitoring of industrial wastewater or as a screening tool for regulators.