Identifying sources of fecal contamination inexpensively with targeted sampling and bacterial source tracking

Most bacterial source tracking (BST) methods are too expensive for most communities to afford. We developed targeted sampling as a prelude to BST to reduce these costs. We combined targeted sampling with three inexpensive BST methods, Enterococcus speciation, detection of the esp gene, and fluorometry, to confirm the sources of fecal contamination to beaches on Georgia's Jekyll and Sea Islands during calm and stormy weather conditions. For Jekyll Island, the most likely source of contamination was bird feces because the percentage of Ent. faecalis was high (30%) and the esp gene was not detected. For the Sea Island beach during calm conditions, the most likely sources of fecal contamination were leaking sewer lines and wildlife feces. The leaking sewer lines were confirmed with fluorometry and detection of the esp gene. For the Sea Island beach during stormflow conditions, the most likely sources of fecal contamination were wildlife feces and runoff discharging from two county-maintained pipes. For the pipes, the most likely source of contamination was bird feces because the percentage of Ent. faecalis was high (30%) and the esp gene was not detected. Sediments were also a reservoir of fecal enterococci for both Jekyll and Sea Islands. Combining targeted sampling with two or more BST methods identified sources of fecal contamination quickly, easily, and inexpensively. This combination was the first time targeted sampling was conducted during stormy conditions, and the first time targeted sampling was combined with enterococcal speciation, detection of the esp gene, and fluorometry.     

Potential of Enterococcus faecalis as a human fecal indicator for microbial source tracking

Regulatory agencies are interested in a fecal indicator bacterium with a host range limited to humans because human fecal contamination represents the greatest hazard to humans, yet is a relatively easy nonpoint source to remedy. Watersheds with human fecal contamination could be given first priority for cleanup. A fecal indicator bacterium with a host range limited to humans and a few other warm-blooded animal species would also simplify microbial source tracking because only a few animal species would be required for any host origin database. The literature suggests that the fecal indicator bacterium Enterococcus faecalis has a limited host range. On this basis, we selected this bacterium for study. Of 583 fecal streptococcal isolates obtained on Enterococcosel agar from Canada goose, cattle, deer, dog, human, chicken, and swine, 392 were considered presumptive enterococci and were subsequently speciated with the API 20 Strep system. Of these isolates, 22 were Ent. durans (5.6%), 61 were Ent. faecalis (15.6%), 98 were Ent. faecium (25.0%), 86 were Ent. gallinarum (21.9%), and 125 were unidentified (31.9%). The host range of the Ent. faecalis isolates was limited to dogs, humans, and chickens. Media were developed to isolate and identify Ent. faecalis quickly from fecal samples and this scheme eliminated Ent. faecalis isolates from dogs. When the remaining Ent. faecalis isolates were ribotyped, it was possible to differentiate clearly among the isolates from human and chicken. It may be that combining the potentially limited host range of Ent. faecalis with ribotyping is useful for prioritizing watersheds with fecal contamination.     

Putative temporal variability of Escherichia coli ribotypes from yearling steers

Escherichia coli is a ubiquitous component of the intestinal microflora of warm-blooded animals, and is an indicator of fecal contamination of surface waters. Ribotype profiling of E. coli is one of several genotypic methods that has been developed to determine the host origin of fecal bacteria. Like most genotypic methods of source tracking, ribotyping requires a host origin database to identify environmental isolates. To determine the extent of temporal variability of ribotypes and its effect on a host origin database, E. coli isolates were obtained from fecal samples of two herds of Black Angus steers at a long-term experimental site at four sampling times from October 1999 to July 2000. Fecal samples were taken from six randomly chosen steers at each time. At a similarity index of 90% as calculated by unweighted pair-group method using arithmetic averages (UPGMA), 240 ribotypes were identified from 451 E. coli isolates. Only 20 ribotypes (8.3%), comprising 33% of the total isolates, were shared among sampling times and were considered resident ribotypes. Two of the twenty resident ribotypes appeared at three sampling times, and the remaining eighteen appeared at two. The majority of the ribotypes, therefore, were transient and unique to each sampling time and steer. Both the apparent turnover of E. coli ribotypes and a clonal diversity index of 0.97 (indicative of extensive ribotype variability) suggest the necessity of ribotyping a large number E. coli isolates per host to establish a host origin database that is independent of temporal variability, or complete enough to be effective.     

Geographic variability of Escherichia coli ribotypes from animals in Idaho and Georgia

Several genotypic methods have been developed for determining the host origin of fecal bacteria in contaminated waters. Some of these methods rely on a host origin database to identify environmental isolates. It is not well understood to what degree these host origin isolates are geographically variable (i.e., cosmopolitan or endemic). This is important because a geographically limited host origin database may or may not be universally applicable. The objective of our study was to use one genotypic method, ribotyping, to determine the geographic variability of the fecal bacterium, Escherichia coli, from one location in Idaho and three locations in Georgia for cattle (Bos taurus), horse (Equus caballus), swine (Sus scrofa), and chicken (Gallus gallus domesticus). A total of 568 fecal E. coli isolates from Kimberly, ID (125 isolates), Athens, GA (210 isolates), Brunswick, GA (102 isolates), and Tifton, GA (131 isolates), yielded 213 ribotypes. The percentage of ribotype sharing within an animal species increased with decreased distance between geographic locations for cattle and horses, but not for swine and chicken. When the E. coli ribotypes among the four host species were compared at one location, the percent of unshared ribotypes was 86, 89, 81, and 79% for Kimberly, Athens, Brunswick, and Tifton, respectively. These data suggest that there is good ribotype separation among host animal species at each location. The ability to match environmental isolates to a host origin database may depend on a large number of environmental and host origin isolates that ideally are not geographically separated.     

ENHANCING FECAL COLIFORM TOTAL MAXIMUM DAILY LOAD MODELS THROUGH BACTERIAL SOURCE TRACKING1

ABSTRACT: Surface water impairment by fecal coliform bacteria is a water quality issue of national scope and importance. In Virginia, more than 400 stream and river segments are on the Commonwealth's 2002 303(d) list because of fecal coliform impairment. Total maximum daily loads (TMDLs) will be developed for most of these listed streams and rivers. Information regarding the major fecal coliform sources that impair surface water quality would enhance the development of effective watershed models and improve TMDLs. Bacterial source tracking (BST) is a recently developed technology for identifying the sources of fecal coliform bacteria and it may be helpful in generating improved TMDLs. Bacterial source tracking was performed, watershed models were developed, and TMDLs were prepared for three streams (Accotink Creek, Christians Creek, and Blacks Run) on Virginia's 303(d) list of impaired waters. Quality assurance of the BST work suggests that these data adequately describe the bacteria sources that are impairing these streams. Initial comparison of simulated bacterial sources with the observed BST data indicated that the fecal coliform sources were represented inaccurately in the initial model simulation. Revised model simulations (based on BST data) appeared to provide a better representation of the sources of fecal coliform bacteria in these three streams. The coupled approach of incorporating BST data into the fecal coliform transport model appears to reduce model uncertainty and should result in an improved TMDL.     

Estrogenic activity in the environment: municipal wastewater effluent, river, ponds, and wetlands

Estrogenic activity of regional water samples was evaluated. Samples obtained from wetlands and ponds involved in various agricultural land uses, from three river sites over four seasons, and from municipal wastewater effluent held in storage lagoons were evaluated. The estrogen-responsive cell line MCF-7 BOS was used in the E-screen assay to determine 17beta-estradiol equivalents (E2 Eq) of water samples extracted by solid-phase extraction. Estrogenic activity in surrounding wetlands and ponds from different land uses was not different, with 10(-12) M E2 Eq (0.3 ppt). Estrogenic activity of Red River samples was within the same range as wetland-pond samples. The highest activity was found downstream from municipal wastewater treatment effluent discharge sites, in winter when river flow was lowest (approximately 6 x 10(-13) M E2 Eq). Results showed that 7 of 20 wetland-pond samples and 5 of 12 river samples were below the limits of quantitation (approximately 3 x 10(-14) M E2 Eq). Toxicity was found in fall and summer river samples upstream from municipal wastewater release sites. The timing of toxicity did not coincide to the presence of elevated fecal coliforms. Estrogenic activity in wastewater effluent from lagoons decreased over time (approximately 25 to 5 x 10(-13) M E2 Eq) with an apparent half-life of 8 d for one lagoon. The median concentration of detectable estrogenic activity in regional water samples was approximately 50-fold less than the median 17beta-estradiol concentration of estradiol detected in some U.S. streams in previous studies.     

Equilibrium sampling used to monitor malodors in a Swine waste lagoon

The concentrations of malodorous compounds in a 0.4-ha anaerobic lagoon that received waste from approximately 2000 sows were monitored during the late summer to late fall of 2006 to gain insight into the factors influencing their concentrations. Selected malodorous compounds were measured by the use of equilibrium samplers consisting of submersible stir plates and stir bar sorbtive sampling with polydimethylsiloxane-coated magnetic stir bars. During the same period, air and water temperatures, suspended solids, total organic carbon and nitrogen content, and wastewater pH were recorded. Concentrations of malodorous compounds were higher at the surface of the lagoon than at the middle or bottom of the lagoon. Skatole concentration, for instance, averaged 54, 24, and 38 microg L(-1) near the surface, in the middle, and at the lowest sampling depths, respectively. While the lagoon was being pumped down during field application of wastewater, concentrations of malodorous compounds fluctuated widely, increased 16-fold as compared with the sampling period before pumping, and continued to increase as fall progressed and temperatures cooled. Suspended solids, volatile suspended solids, and total organic carbon increased near the bottom of the lagoon during this same period. The increases in the concentrations of malodorous compounds in the wastewater during the fall could have been due to a combination of several factors. These factors include reduced degradation by lagoon bacteria, less wind stripping of volatile compounds from the lagoon surface due to lowering of the lagoon surface after crop application, and/or reduced evaporation of malodorous compounds due to falling temperatures.     

Evaluating sewage-associated JCV and BKV polyomaviruses for sourcing human fecal pollution in a coastal river in Southeast Queensland, Australia

In this study, the host-sensitivity and host-specificity of JC virus (JCV) and BK virus (BKV) polyomaviruses were evaluated by testing wastewater and fecal samples from nine host groups in Southeast Queensland, Australia. The JCV and BKV polyomaviruses were detected in 63 human wastewater samples collected from primary and secondary effluent, suggesting high sensitivity of these viruses in human wastewater. In the 81 animal wastewater and fecal samples tested, 80 were polymerase chain reaction (PCR) negative for the JCV and BKV markers. Only one sample (out of 81 animal wastewater and fecal samples) from pig wastewater was positive. Nonetheless, the overall host-specificity of these viruses to differentiate between human and animal wastewater and fecal samples was 0.99. To our knowledge, this is the first study in Australia that reports on the high specificity of JCV and BKV polyomaviruses. To evaluate the field application of these viral markers for detecting human fecal pollution, 20 environmental samples were collected from a coastal river. In the 20 samples tested, 15% (3/20) and 70% (14/20) samples exceeded the regulatory guidelines for Escherichia coli and enterococci levels for marine waters. In all, five (25%) samples were PCR positive for JCV and BKV, indicating the presence of human fecal pollution in the coastal river investigated. The results suggest that JCV and BKV detection using PCR could be a useful tool for identifying human-sourced fecal pollution in coastal waters.     

Use of Escherichia coli BOX-PCR fingerprints to identify sources of fecal contamination of water bodies in the State of São Paulo, Brazil

Repetitive element sequence-based polymerase chain reaction (rep-PCR) is one of the commonest methods used to identify sources of fecal contamination of water systems. In this work, BOX-A1R-based repetitive extragenic palindromic-PCR (BOX-PCR) was used to discriminate Escherichia coli strains originating from different animals and water sources, and the suitability of the technique for bacterial source tracking (BST) was evaluated. A total of 214 strains from humans, 150 strains from animals, 55 strains from sewage and 77 strains from water bodies were analyzed by the BOX-PCR technique. When maximum similarity between the fingerprints was used, a correct classification rate of 84% was achieved for strains from human and animal sources. Furthermore, 95% of the strains found in sewage were classified as being from human sources by at least one of the four classification tools used. Classification of the strains found in water bodies in the State of S?o Paulo was based on the fingerprints obtained for human and animal sources. Most of the sampling sites appeared to be affected by mixed sources of fecal contamination. The use of BOX-PCR for BST could be especially valuable in developing countries, where simplicity and cost are important considerations.     

Respective contributions of point and non-point sources of E. coli and enterococci in a large urbanized watershed (the Seine river, France)

Because the large rivers of the Seine watershed have a low microbiological water quality, the main sources of fecal contamination were investigated in the present study. The inputs of the point (wastewater treatment plants (WWTPs) effluents) and non-point sources (surface runoff and soil leaching) of fecal bacteria were quantified for Escherichia coli and intestinal enteroccoci used as bacterial indicators. In order to assess the contamination through non-point sources, fecal indicators abundance was estimated in samples collected in small streams located in rural areas upstream from all point sources; these small rivers were characterized by the land use of their watershed. Bacterial indicator numbers were also measured in effluents of WWTPs, some using classical treatment (settling followed by activated sludge process) and some using an additional disinfection stage (UV irradiation). These data were used to estimate the respective importance of each type of source at the scale of the whole Seine river watershed taking into account the land use and the population density. It shows the predominant importance of the point sources of fecal indicator bacteria at the scale of the whole watershed. In a scenario in which activated sludge treatment would be complemented with UV in all WWTPs located in this watershed, the non-point sources of fecal indicator bacteria would be dominant.     

Evaluating slurry broadcasting and injection to ley for phosphorus losses and fecal microorganisms in surface runoff

The recent growth in the size of dairy cattle farms and the concentration of farms into smaller areas in Finland may increase local water pollution due to increased manure production and slurry application to grass. Therefore, a field study was conducted to monitor losses of total phosphorus (TP), dissolved reactive phosphorus (DRP), and fecal microorganisms in surface runoff from a perennial ley. Cattle slurry was added once a year in June 1996-1997 (Study I) and biannually in June and October 1998-2000 (Study II). The slurry was surface broadcast or injected into the clay soil. The field had a slope of 0.9 to 1.7%. Mineral fertilizer was applied on control plots. Biannual slurry broadcasting increased DRP (p < 0.001) and TP losses (p < 0.001) and numbers of fecal microorganisms in surface runoff waters. The highest losses of TP (2.7 kg ha(-1) yr(-1)) and DRP (2.2 kg ha(-1) yr(-1)) and the highest numbers of fecal coliforms (880 colony-forming units [CFU] per 100 mL) and somatic coliphages (2700 plaque-forming units [PFU] per 100 mL) were measured after broadcasting slurry to wet soil followed by rainfall in fall 1998. Injection reduced the TP and DRP losses in surface runoff by 79 and 86%, respectively, compared with broadcasting (17 Oct. 1998-27 Oct. 1999). Corresponding numbers for fecal coliforms were 350 CFU (100 mL)(-1) and for somatic coliphages were 110 PFU (100 mL)(-1) in surface runoff after injection in October 1998. Slurry injection should be favored when spreading slurry amendments to grassland to avoid losses of P and fecal microorganisms in runoff to surface waters.     

Reduction of pathogen indicator organisms in dairy wastewater using an ecological treatment system

Ecological treatment systems can provide a sustainable, plant-based alternative to traditional wastewater treatment. One factor essential to the success of these systems is ensuring their ability to reduce coliform concentrations in wastewater. Wastewater is the primary source of fecal contamination in aquatic ecosystems, containing total and fecal coliforms on the order of 10(8)-10(10) and 10(7)-10(9) CFU L(-1), respectively. This study assessed the ability of an ecological treatment system to reduce concentrations of total coliforms and Escherichia coli from dairy wastewater. Low strength wastewater was pumped into the system during July of 2005 and high strength in September 2005. Wastewater passes through a series of anaerobic, aerobic, and clarifier reactors and wetland cells before exiting the system. Regardless of wastewater strength, average total coliform and E. coli concentrations were consistently reduced by at least 99% from influent to effluent, with the majority of the reduction (76%) occurring in the first two reactors. Relationships between internal concentrations of solids and coliforms indicated that increased reduction of solids may further reduce coliform concentrations. Although U.S. Environmental Protection Agency discharge requirements for E. coli were not always met, the substantial reductions achieved indicate that ecological treatment systems have the potential to successfully reduce coliforms in wastewater to meet discharge limits. The results from this study will be used to guide design and management of future ecological treatment systems, so that larger and more consistent coliform reductions can be achieved.     

AGRICULTURAL LAND USE IMPACTS ON BACTERIAL WATER QUALITY IN A KARST GROUNDWATER AQUIFER1

ABSTRACT: The impact on water quality by agricultural activity in karst terrain is an important consideration for resource management within the Appalachian Region. Karst areas comprise about 18 percent of the Region's land area. An estimated one-third of the Region's farms, cattle, and agricultural market value are on karst terrain. The purpose of this study was to compare fecal bacteria densities in karst groundwater impacted by two primary agricultural land uses in central Appalachia. Fecal bacteria densities were measured in cave streams draining two primary land management areas. The first area was pasture serving a beef cow-calf operation. The second area was a dairy. Neither area had best management practices in place for controlling animal wastes. Median fecal coliform and fecal streptococcus densities were highest in cave streams draining the dairy. Median fecal coliform densities in the dairy-impacted stream were greater than 4,000 CFU/100 ml and the median fecal coliform densities in the pasture-impacted streams were less than 10 CFU/100 ml. Median fecal streptococcus densities in the same streams were greater than 2,000 CFU/100 ml and 32 CFU/100 nil, respectively. A second dairy, with best management practices for control of animal and milkhouse waste, did not appear to be contributing significant amounts of fecal bacteria to the karst aquifer. It was concluded that agriculture was affecting bacterial densities in the karst aquifer. New management practices specifically designed to protect karst groundwater resources may be one way to protect the groundwater resource.     

MULTIOBJECTWE ALLOCATION OF WATER SHORTAGE IN THE SVARTA RIVER,SWEDEN1

ABSTRACT: A model is proposed for allocation of water shortages among competing water uses in the Svarta River basin in Sweden. The three major competing uses in the basin are hydroelectricity generation, irrigation water supply, and urban water supply. Minor uses that impact upon the allocation are minimum river flow requirements for fishlife and for dilution of treated wastewater, and storage level restrictions for recreation purposes in the main storage facility, Lake Sommen. Analysis of the competing demands on the water are modeled through the method-of-weights multiobjective technique using a deterministic mixed-integer optimization formulation. The (0–1) variables in the formulation are required to synthesize the restricted validity of permits for withdrawal of irrigation water from the river and to simulate the complex operating rules of the major regulation facility on the river. Due to the deterministic nature of the formulation, the model is used on a hydrologic scenario basis. Use of the model is demonstrated by application to the Svarta River.     

Reduction of malodorous compounds from a treated swine anaerobic lagoon

There is a need for treatment technologies that can eliminate environmental problems associated with anaerobic lagoons. These technologies must be able to capture nutrients, kill pathogens, and reduce emissions of ammonia and nuisance odors. To meet these needs, a full-scale wastewater treatment plant was installed as a demonstration project on one of three 4360-pig (Sus scrofa) production units in a finishing farm in Duplin County, North Carolina. Once the treatment plant was operational, flow of raw manure into the unit's corresponding lagoon was discontinued and the lagoon was used to store treated wastewater. Water quality was monitored in the converted lagoon and in the two conventional lagoons. A gas chromatographic method was developed to measure concentration of five selected malodorous compounds (phenol, p-cresol, 4-ethylphenol, indole, and skatole) in liquid lagoon samples. Dramatic improvements in the water quality parameters TKN, NH3-N, solids, COD, and BOD in the converted waste lagoon paralleled reductions in malodorous compounds. Nine months after conversion, identified malodorous compounds in liquid extractions averaged 6.6 and 38.8 ng mL(-1) in water from the converted lagoon and the conventional lagoons, respectively. The reduction was particularly marked for p-cresol, 4-ethylphenol, and skatole, all of which make important contributions to swine waste odors due to their characteristic odors and low detection thresholds.     

PREDICTING FECAL COLIFORM BACTERIA LEVELS IN THE CHARLES RIVER,MASSACHUSETTS, USA1

In Massachusetts, the Charles River Watershed Association conducts a regular water quality monitoring and public notification program in the Charles River Basin during the recreational season to inform users of the river's health. This program has relied on laboratory analyses of river samples for fecal coliform bacteria levels, however, results are not available until at least 24 hours after sampling. To avoid the need for laboratory analyses, ordinary least squares (OLS) and logistic regression models were developed to predict fecal coliform bacteria concentrations and the probabilities of exceeding the Massachusetts secondary contact recreation standard for bacteria based on meteorological conditions and streamflow. The OLS models resulted in adjusted R²s ranging from 50 to 60 percent. An uncertainty analysis reveals that of the total variability of fecal coliform bacteria concentrations, 45 percent is explained by the OLS regression model, 15 percent is explained by both measurement and space sampling error, and 40 percent is explained by time sampling error. Higher accuracy in future bacteria forecasting models would likely result from reductions in laboratory measurement errors and improved sampling designs.     

A MODELLING METHOD FOR EVALUATING WATER QUALITY POLICIES IN NONSERIAL RIVER SYSTEMS1

ABSTRACT: This paper describes a modelling method which simplifies the evaluation of water quality policies for nonserial (branching) river systems. The method introduces dummy facilities at the junctions of the major tributary branches with the mainstream as replacements for the facilities and activities on the tributaries. The cost functions for the dummy facilities and the DO and BOD concentrations at the junctions as determined for each tributary are introduced into the mainstream serial system model which is then solved for the optimal values of the mainstream treatment plant efficiencies, the dummy facility effeciencies, and the tributary system DO and BOD concentrations using nonlinear programming. Given the optimum values for the dummy facility efficiencies and the values for the tributary system DO and BOD concentrations, the optimum values for the tributary treatment plant efficiencies are found using nonlinear programming. The method is applied to a river system with a mainstream and one major tributary which contain industrial and municipal organic and thermal polluters and their associated wastewater treatment plants.     

Source characterization of nervous system active pharmaceutical ingredients in healthcare facility wastewaters

Nervous system active pharmaceutical ingredients (APIs), including anti-depressants and opioids, are important clinically administered pharmaceuticals within healthcare facilities. This study provides source characterization data describing the composition and magnitude of nervous system APIs present in healthcare facility wastewaters. Concentrations and mass loadings of ten nervous system APIs and three nervous system API metabolites are reported for wastewaters from a hospital, nursing, assisted living, and independent living facility within a single municipality. Concentrations of nervous system APIs ranged from non-detectable levels for alprazolam in all four facility wastewaters to a high of 290 ng/L amitriptyline in nursing facility wastewater. The summed mean concentration of all thirteen analytes ranged from 402 ng/L in independent living facility wastewater to 624 ng/L in assisted living facility wastewater. Wastewater flow rates from each facility were combined with concentration data to estimate the daily mass loading of nervous system APIs leaving each facility through wastewater discharge to the municipal sewer system. The total mass loading of all thirteen analytes for the hospital, nursing, assisted living, and independent living facility was 228, 44, 29.5, and 28.1 mg/day, respectively. The total mass loading of nervous system APIs contributed to the municipality's wastewater from all four facilities was 330 mg/day.     

Denitrification in anaerobic lagoons used to treat swine wastewater

Anaerobic lagoons are commonly used for the treatment of swine wastewater. Although these lagoons were once thought to be relatively simple, their physical, chemical, and biological processes are very complex. This study of anaerobic lagoons had two objectives: (i) to quantify denitrification enzyme activity (DEA) and (ii) to evaluate the influence of lagoon characteristics on the DEA. The DEA was measured by the acetylene inhibition method. Wastewater samples and physical and chemical measurements were taken from the wastewater column of nine anaerobic swine lagoons from May 2006 to May 2009. These lagoons were typical for anaerobic swine lagoons in the Carolinas relative to their size, operation, and chemical and physical characteristics. Their mean value for DEA was 87 mg N2O-N m(-3) d(-1). In a lagoon with 2-m depth, this rate of DEA would be compatible with 1.74 kg N ha(-1) d(-1) When nonlimiting nitrate was added, the highest DEA was compatible with 4.38 kg N ha(-1) d(-1) loss. Using stepwise regression for this treatment, the lagoon characteristics (i.e., soluble organic carbon, total nitrogen, temperature, and NO3-N) provided a final step model R2 of 0.69. Nitrous oxide from incomplete denitrification was not a significant part of the system nitrogen balance. Although alternate pathways of denitrification may exist within or beneath the wastewater column, this paper documents the lack of sufficient denitrification enzyme activity within the wastewater column of these anaerobic lagoons to support large N2 gas losses via classical nitrification and denitrification.     

Long-term performance of a land treatment facility for the bioremediation of non-volatile oily wastes

The current study reports on operational and performance aspects of a land treatment facility for managing oily wastes generated from heavy vehicle maintenance over a 5-year period. Samples of soil and groundwater from the land treatment plots were analyzed for a range of contaminants and microorganisms over this period. The soil analyses indicated that the process has been operating efficiently even at high wastewater loadings with maximum degradation rates of 250 mg/kg per day (year 1) and average rates of 10–35 mg/kg per day (years 2–5). Hydrocarbon degraders comprised more than 80% of the total (measured) soil heterotrophic population and were present at 10⁶–10⁸ (per gram soil) once the process was optimized. The facility was capable of treating 140 kl of oily wastewater per day (average petroleum hydrocarbon content of 2% w/v) over the entire period. During this time there was no evidence of accumulation of any major metals or polycyclic aromatic hydrocarbon (PAH) compounds in the soil. Groundwater sampling and analysis indicated that the land treatment facility was not leading to contamination of any groundwater taken from bores in the vicinity of the facility. The land treatment process continues to be effective for treatment of non-volatile waste oils at this remote and high evaporation (arid) site.