Development and Integration of Quantitative Real-Time PCR Methods for Detection of Mitochondrial DNA and Methanobrevibacter smithii nifH Gene as Novel Microbial Source Tracking Tools

A novel microbial source tracking (MST) method based on the detection of human and non-human markers was developed and applied to track the origin of fecal pollution in water systems. Mitochondrial DNA sequences were used to develop new quantitative real-time polymerase chain reaction (qPCR) assays for dog, poultry, and gull. The targets were included as part of a toolbox including human, cow, pig, and sheep assays. A primer and probe set for the detection of the human-specific nifH gene of Methanobrevibacter smithii was also designed as an indicator of human fecal contamination. The assays were tested for specificity and applied to fecal-spiked surface waters and environmental samples collected from two river catchments impacted by sources of human and non-human fecal contamination. The MST methods described were applicable to both spiked waters and environmental samples, and using the two approaches the origin of fecal pollution could be successfully determined in mixed source fecally polluted waters.     

Analysis of the life cycle of non-ferrous minerals

This paper examines the utility of a methodology for forecasting the lifecycle of metals obtained from non-renewable resources which are subject to high rates of growth in consumption. A rate method is used, subject to arbitrarily chosen upper limits of reserves. One general pattern which emerges is that extraction of the resource will tend to rise exponentially in face of falling rate of discovery of new reserves; a degree of ‘overshoot’ occurs. In other words, an economy might continue to be geared to a given non-renewable resource with a degree of optimism about reserve life being maintained until well after it is justifiable and there may be inadequate time for the price mechanism alone to operate to generate development of substitutes, and for the introduction of intensive recycle technology or greater selectivity and economies in the use of the original resource.It is concluded that the methodology used here could be useful to study the possible forms of lifecycles for the base metals.     

Transport of lead and diesel fuel through a peat soil near Juneau, AK: a pilot study

A set of peat column experiments was used to determine the transport potential of lead (Pb) and diesel range organics (DRO) in palustrine slope wetlands near Juneau, AK. This project is important to southeast Alaskan communities because limited land resources are forcing development of regional wetlands. This study was instigated by concerns that proposed modifications to a nearby rifle range using DRO-contaminated soil posed a potential risk to an anadromous fish-bearing stream 250 m from the site. Three pairs of peat columns were extracted from the rifle range for analysis, one pair along and two pairs across the natural bedding planes of the soil. One column in each pair was spiked with Pb and DRO and the other was used as a control. Approximately 1-year worth of water (171 cm) was passed through each column and leachate was collected at regular intervals. The results showed that substantial DRO transport only occurred along the bedding planes. Leads was surprisingly mobile, both along and across the bedding planes with estimated soil-water partition coefficients several orders of magnitude lower than commonly published values, probably because the peat was heavily Pb-loaded by lead from bullets and because the peat's acidic, organic-rich environment enhanced Pb mobility. The chemical outflow behavior agreed with a simple macropore transport model. These results underscore the need for caution when developing regional wetlands.     

Cesium migration in Hanford sediment: a multisite cation exchange model based on laboratory transport experiments

Cs+ transport experiments carried out in columns packed with uncontaminated Hanford formation sediment from the SX tank farm provide strong support for the use of a multisite, multicomponent cation exchange model to describe Cs+ migration in the Hanford vadose zone. The experimental results indicate a strong dependence of the effective Cs+ Kd on the concentrations of other cations, including Na+ that is present at high to extremely high concentrations in fluids leaking from the Hanford SX tanks. A strong dependence of the Cs+ Kd on the aqueous Cs+ concentration is also apparent, with retardation of Cs+ increasing from a value of 41 at a Cs+ concentration of 10(-4) M in the feed solution to as much as 282 at a Cs+ concentration of 5x10(-7) M, all in a background of 1 M NaNO3. The total cation exchange capacity (CEC) of the Hanford sediment was determined using 22Na isotopic equilibrium exchange in a flow-through column experiment. The value for the CEC of 120 microeq/g determined with this method is compatible with a value of 121.9 microeq/g determined by multi-cation elution. While two distinct exchange sites were proposed by Zachara et al. [Geochim. Cosmochim. Acta 66 (2002) 193] based on binary batch exchange experiments, a third site is proposed in this study to improve the fit of the Cs+-Na+ and Cs+-Ca+ exchange data and to capture self-sharpened Cs+ breakthrough curves at low concentrations of Cs+. Two of the proposed exchange sites represent frayed edge sites (FES) on weathered micas and constitute 0.02% and 0.22% of the total CEC. Both of the FES show a very strong selectivity for Cs+ over Na+ (K(Na-Cs)=10(7.22) and 10(4.93), respectively). The third site, accounting for over 99% of the total CEC, is associated with planar sites on expansible clays and shows a smaller Na+-Cs+ selectivity coefficient of 10(1.99). Parameters derived from a fit of binary batch experiments alone tend to under predict Cs+ retardation in the column experiments. The transport experiments indicate 72-90% of the Cs+ sorbed in experiments targeting exchange on FES was desorbed over a 10- and 24-day period, respectively. At high Cs+ concentrations, where sorption is controlled primarily by exchange on planar sites, 95% of the Cs+ desorption was desorbed. Most of the difficulty in desorbing Cs+ from FES is a result of the extremely high selectivity of these sites for Cs+, although truly irreversible sorption as high as 23% was suggested in one experiment. The conclusion that Cs+ exchange is largely reversible in a thermodynamic sense is supported by the ability to match Cs+ desorption curves almost quantitatively with an equilibrium reactive transport simulation. The model for Cs+ retardation developed here qualitatively explains the behavior of Cs+ in the Hanford vadose zone underneath a variety of leaking tanks with differing salt concentrations. The high selectivity of FES for Cs+ implies that future desorption and migration is very unlikely to occur under natural recharge conditions.     

Toxicity of copper in sewage sludge

Sewage sludge is a source of organic matter and nutrients, but a major obstacle for its recycling is that the municipal wastewater sludge has low but significant levels of contaminants. This investigation, on the acute toxicity of copper in sewage sludge, was conducted with three organisms, Daphnia magna, Lemna minor and Raphanus sativus (seeds). The toxicity of the leakage water from sewage sludge spiked with CuSO(4) was studied for 64 days. The toxicity increased during the first 8-16 days and then started to decrease. The first increase in toxicity was due to ammonia, but after 32 days, a dose-related effect of copper was found. After 64 days, L. minor had an EC50 of 3800 mg Cu/kg dw for 7 days growth inhibition, a LOEC of 3200 mg Cu/kg dw and a NOEC of 1600 mg Cu/kg dw. D. magna had an EC50 of 18100 mg Cu/kg dw (24-h immobility) and a NOEC of 12800 mg Cu/kg dw. Root elongation of R. sativus was reduced at 25600 mg Cu/kg dw. Both for Daphnia and Lemna, the pH of the leakage water had an effect of the toxicity. This means that chemical speciation and bioavailability is very important for the hazard assessment of copper in sludge and soil.     

AN EXAMINATION OF RELATIVE PERMEABILITY RELATIONS FOR TWO-PHASE FLOW IN POROUS MEDIA1

ABSTRACT: A key parameter in modeling two-phase flow phenomena is relative permeability. It is important to understand which variables influence relative permeability, especially since so few measurements of relative permeability have been made for typical contaminants at hazardous waste sites. This paper focuses on the effect of five variables on relative permeability: intrinsic permeability, pore-size distribution, viscosity ratio, interfacial tension, and wettability, by critically reviewing previously published relative permeability experiments. The wide variability in the functional relationship between relative permeability and saturation should be considered in attempts to model two-phase flow.     

Occurrence of potential bacterial pathogens in coastal areas of the Adriatic Sea

A monitoring program was carried out in bathing waters along the sea-coast of the Adriatic Sea. The investigation was aimedat obtaining a quali-quantitative picture of microorganisms ofanthropic origin and of a more strictly autochthonous origin and at verifying the existence of possible reciprocal correlations and combinations of relationship between them. Results showed that the concentrations of indicators of faecal contamination, whether within or over the values established by legislation, are not always related to the presence and density of microorganisms of environmental origin but the regression modelsused can help to describe how these latter microorganisms can bein relation to levels of the other parameters. The difficultyis evident of the predictability of health effects swimming-associated; for this reason more complex models that are currently used should be taken into account for minimising the risk for bathers.     

Associating Ecosystem Service Losses with Indicators of Toxicity in Habitat Equivalency Analysis

Habitat equivalency analysis (HEA) was developed as a tool to scale mitigation or restoration when habitat is contaminated by hazardous substances or has been otherwise harmed by anthropogenic activities. Applying HEA involves balancing reductions in habitat quality against gains from restoration actions, and quantifying changes in habitat quality in terms of ecological services. We propose a framework for developing ecological service definitions and measures that incorporate knowledge about the impacts of chemical contaminants on biota. We describe a general model for integrating multiple lines of evidence about the toxicity of hazardous substances to allow mapping of toxicological inputs to ecological service losses. We provide an example of how this framework might be used in a HEA that quantifies ecological services provided by estuarine sediments contaminated with polycyclic aromatic hydrocarbons.