Influence of a soil enzyme on iron-cyanide complex speciation and mineral adsorption

Cyanide is commonly found as ferrocyanide [Fe(II)(CN)(6)](-4) and in the more mobile form, ferricyanide [Fe(III)(CN)(6)](-3) in contaminated soils and sediments. Although soil minerals may influence ferrocyanide speciation, and thus mobility, the possible influence of soil enzymes has not been examined. In a series of experiments conducted under a range of soil-like conditions, laccase, a phenoloxidase enzyme derived from the fungi Trametes versicolor, was found to exert a large influence on iron-cyanide speciation and mobility. In the presence of laccase, up to 93% of ferrocyanide (36-362ppm) was oxidized to ferricyanide within 4h. No significant effect of pH (3.6 and 6.2) or initial ferrocyanide concentration on the extent or rate of oxidation was found and ferrocyanide oxidation did not occur in the absence of laccase. Relative to iron-cyanide-mineral systems without laccase, ferrocyanide adsorption to aluminum hydroxide and montmorillonite decreased in the presence of laccase and was similar to or somewhat greater than that of ferricyanide without laccase. Laccase-catalyzed conversion of ferrocyanide to ferricyanide was extensive though up to 33% of the enzyme was mineral-bound. These results demonstrate that soil enzymes can play a major role in ferrocyanide speciation and mobility. Biotic soil components must be considered as highly effective oxidation catalysts that may alter the mobility of metals and metal complexes in soil. Immobilized enzymes should also be considered for use in soil metal remediation efforts.     

Lability of polycyclic aromatic hydrocarbons in the rhizosphere

Remediation of soils containing high concentrations of polycyclic aromatic hydrocarbons (PAHs) seldom results in complete removal of contaminants, but residual toxicity often is reduced. In this study, soil from a former manufactured gas plant site was treated for 12 months by phytoremediation and then tested for total PAHs, Tenax-TA extractable ("labile") PAHs, aqueous soluble PAHs (PAH(wp)) , and biotoxicity assessed by earthworms survival, nematode mortality, emergence of lettuce seedlings, and microbial respiration. Prior to phytoremediation, the soil had toxic impacts on all bioassays (except the nematodes), and 12 months of remediation decreased this response. Change in labile PAHs was a predictor for change in total PAH for 3- and 4-ring compounds but not for the 5- and 6-ring. Decreases in labile PAHs were correlated (r(2)>or=0.80) with toxicity in the bioassays except microbial respiration. PAH(wp) was correlated only with nematode toxicity prior to remediation but with none of the tests after remediation. Total PAHs were not correlated with any of the bioassay tests. Tenax-TA appears to have potential for predicting residual toxicity in remediated soils and is superior to total concentrations for that application.     

In vitro bioassay for reactive toxicity towards proteins implemented for water quality monitoring

Reactive organic chemicals comprise a large number of compounds with a variety of reactive moieties. While most assays for reactive toxicity focus on DNA damage, reactivity towards proteins can also lead to irreparable damage, but reactivity towards proteins is typically not included in any test battery for water quality assessment. Glutathione (GSH) is a small tripeptide whose cysteine moiety can serve as a model for nucleophilic sites on proteins. GSH is also an important indicator of detoxification processes and the redox status of cells and due to its protective role, depletion of GSH ultimately leads to adverse effects. A bioassay based on genetically modified Escherichia coli strains was used to quantify the specific reactivity towards the protein-like biological nucelophile GSH. The significance of GSH for detoxification was assessed by comparing the growth inhibition induced by reference chemicals or water samples in a GSH-deficient strain to its fully functional parent strain. The GSH deficient strain showed the same sensitivity as the GSH proficient strain to non-reactive and DNA damaging chemicals, but was more sensitive to chemicals that attack cysteine in proteins. The difference in effect concentrations for 50% inhibition of growth assessed as biomass increase (EC(50)) between the two strains indicates the relevance of GSH conjugation as a detoxification step as well as direct reactivity with cysteine-containing proteins. Seven reference compounds serving as positive and negative controls were investigated. The E. coli strain that lacks GSH was four times more sensitive towards the positive control Sea-Nine, while negative controls benzo[a]pyrene, 2-aminoanthracene, phenol, t-butylhydroquinone, methyl methane sulfonate and 4-nitroquinoline oxide showed equal effect concentrations in both strains. Water samples collected across an indirect potable reuse scheme representing the complete water cycle from sewage to drinking water in South East Queensland, Australia were used to evaluate the applicability of the E. coli assay for reactive toxicity in water samples. While the EC(50) values of the GSH+ strain showed similar trends as in other biological endpoints over the various treatment chains, the specific response indicative of protein damage was only observed in samples that had undergone chlorination as a disinfection process. High natural organic matter or other matrix components disturbed the bioassay so much that we recommend it for future routine testing only in tertiary treated water or drinking water.     

Spatial and Temporal Patterns in Fish Assemblages Following an Artificially Extended Floodplain Inundation Event, Northern Murray-Darling Basin, Australia

Water extraction from dryland rivers is often associated with declines in the health of river and floodplain ecosystems due to reduced flooding frequency and extent of floodplain inundation. Following moderate flooding in early 2008 in the Narran River, Murray-Darling Basin, Australia, 10,423 ML of water was purchased from agricultural water users and delivered to the river to prolong inundation of its terminal lake system to improve the recruitment success of colonial waterbirds that had started breeding in response to the initial flooding. This study examined the spatial and temporal patterns of fish assemblages in river and floodplain habitats over eight months following flooding to assess the possible ecological benefits of flood extension. Although the abundances of most fish species were greater in river channel habitats, the fish assemblage used floodplain habitats when inundated. Young-of-the-year (4–12 months age) golden perch (Macquaria ambigua) and bony bream (Nematalosa erebi) were consistently sampled in floodplain sites when inundated, suggesting that the floodplain provides rearing habitat for these species. Significant differences in the abundances of fish populations between reaches upstream and downstream of a weir in the main river channel indicates that the effectiveness of the environmental water release was limited by restricted connectivity within the broader catchment. Although the seasonal timing of flood extension may have coincided with sub-optimal primary production, the use of the environmental water purchase is likely to have promoted recruitment of fish populations by providing greater access to floodplain nursery habitats, thereby improving the ability to persist during years of little or no flow.     

Assessing the Response of Estuarine Intertidal Assemblages to Urbanised Catchment Discharge

Measurement of intertidal rock assemblages was investigated as a potential biological indicator to provide a quantitative estimate of the impact of urbanised catchment discharge on the estuaries of Sydney, Australia, from 1995 to 1999. Based on the presence and characteristics of adjacent human activities, sampling locations were categorised as: Bush; Urban; Urban with Sewer Overflows; and Industry with Sewer Overflows. In Sydney Harbour, variation in assemblage structure was measured between most impact categories, however differences between impact categories were not consistent for each year. Nevertheless, in years of above average rainfall (1998–1999), reference assemblages adjacent to national parks and distant from urbanisation were different to all other putatively impacted assemblages. Variability within assemblages was least at reference locations in each year and greatest at locations adjacent to stormwater canals and sewer overflows, particularly in 1998–1999. Variation in assemblage structure in Sydney estuaries was most strongly correlated with chlorophyll – a concentrations. Univariate analysis also identified highly significant differences for a number of factors, however, interactions between year, impact categories and location for numerous analyses, confounded the differentiation between impact categories. The results suggest that intertidal rock assemblages in Sydney Harbour and surrounding estuaries appear to be responding to the quality and quantity of discharge from urbanised catchments and, furthermore, that assemblages are more suitable than individual taxa to indicate the difference between Bush and anthropogenically disturbed estuarine locations.     

Characteristics in oxidative degradation by ozone for saturated hydrocarbons in soil contaminated with diesel fuel

Although alkanes are relatively less reactive to chemical oxidation compared to alkenes, the chemical oxidation of alkanes has not been adequately explored in the context of environmental remediation efforts. Laboratory-scale column experiments were therefore conducted with soil artificially contaminated by diesel fuel as a surrogate for alkanes of environmental relevance. Particular attention was paid to saturated hydrocarbons refractory to volatilization. Reaction conditions involve 1485mgkg(-1) of the initial concentration of diesel range organics (DRO) and a constant ozone concentration of 119+/-6mgl(-1) at the flow rate of 50mlmin(-1). The observed removal of DRO reached 94% over 14h of continuous ozone injection. Ozone oxidation demonstrated effective removal of non-volatile DRO in the range of C(12)-C(24). Each alkane compound displayed comparable degradation kinetics, suggesting virtually no selectivity of ozone reactions with alkanes in soil. A pseudo-first order kinetic model closely simulated the removal kinetics, yielding a reaction rate constant of 0.213 (+/-0.021)h(-1) and a half-life of 3.3 (+/-0.3)h under the experimental conditions used in this study. An estimate of ozone demand was 32mg of O(3) (mgDRO)(-1).