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Numerous harmful chemicals are introduced every year in the environment through anthropogenic and geological activities raising global concerns of their ecotoxicological effects and decontamination strategies. Biochar technology has been recognized as an important pillar for recycling of biomass, contributing to the carbon capture and bioenergy industries, and remediation of contaminated soil, sediments and water. This paper aims to critically review the application potential of biochar with a special focus on the synergistic and antagonistic effects on contaminant-degrading microorganisms in single and mixed-contaminated systems. Owing to the high specific surface area, porous structure, and compatible surface chemistry, biochar can support the proliferation and activity of contaminant-degrading microorganisms. A combination of biochar and microorganisms to remove a variety of contaminants has gained popularity in recent years alongside traditional chemical and physical remediation technologies. The microbial compatibility of biochar can be improved by optimizing the surface parameters so that toxic pollutant release is minimized, biofilm formation is encouraged, and microbial populations are enhanced. Biocompatible biochar thus shows potential in the bioremediation of organic contaminants by harboring microbial populations, releasing contaminant-degrading enzymes, and protecting beneficial microorganisms from immediate toxicity of surrounding contaminants. This review recommends that biochar-microorganism co-deployment holds a great potential for the removal of contaminants thereby reducing the risk of organic contaminants to human and environmental health Refereed/Peer-reviewed

The addition of wood fly ash to an agrowaste anaerobic digestate has been proposed for improving the carbon to nutrient ratio of the soil organic amendment (C/N/P < 100/10/1). The ash-based treatment also aimed to improve properties of the anaerobic digestate as controlled-release fertilizer by decreasing the availability of nitrogen, carbon, and phosphorus. Treated (39.86 ± 0.94 g) and untreated (33.51 ± 0.84 g) digestates were incubated for 7 h at 20 °C and 100 rpm, in a 250-mL chamber with a 0.11 M sulfuric acid trap of 4.38 ± 0.02 mL. By the end of the incubation, the blend of 13.12 ± 2.87% ash and 86.88 ± 2.87% digestate released 8.95 ± 2.19 times more ammonia than the digestate alone. This was in agreement with the lower content of water-soluble ammoniacal nitrogen in the treated digestate (93.35 ± 21.00 mg/kg) than in the digestate incubated without the addition of the ash (357.64 ± 54.10 mg/kg). As per the very low levels of water-soluble nitric nitrogen found in the treated (0.16 ± 0.05 mg/kg) and the untreated (0.06 ± 0.02 mg/kg) digestate, the emissions of nitrous oxide were discarded. The availability of carbon increased 1.43 ± 0.82 times and the availability of phosphorus decreased approximately 100 times due to the addition of the ash to the digestate. The ash-based treatment was successful in improving the C/N/P of the digestate from 121.51 ± 57.18/9.94 ± 4.21/1 to 42.05 ± 9.88/2.48 ± 0.58/1.

Fluorene, a low molecular weight polycyclic aromatic hydrocarbon (PAH), is of immense environmental interest because of its carcinogenicity, teratogenicity, mutagenicity, toxicity and persistence to microbial degradation. Existentially, there is paucity of information on PAH degradation by fungi isolated from marine environment. Therefore, this study investigated fluorene degradation efficiency of marine derived filamentous fungus, Mucor irregularis strain bpo1 (GenBank Accession Number: MK373020). Response Surface Methodology (RSM) using Box–Behnken Design (BBD) was successfully deployed in the optimization of process parameters (pH-7, temperature-32.5 °C, substrate concentration-100 mg L−1 and dry weight-2 g) resulting in 81.50% fluorene degradation on 5th day. The design and regression model were found to be statistically significant, adequate and appropriate with p < 0.0001, F value = 202.39, and predicted coefficient of determination (R2 = 0.9991). Optimization of the vital constituents of the mineral salt medium (MSM) used for the study using RSM-Central Composite Design (CCD) resulted in 79.80% fluorene degradation rate. Enhanced fluorene degradation efficiency (82.50%) was recorded when the optimized process variables were subjected to growth-linked validation experiments. The enzyme activities revealed 87%, 59% and 31% induction of laccase, manganese peroxidase and lignin peroxidase respectively. Four metabolites; 9H-fluoren-9-one, benzene-1,2-dicarboxylic acid, 2-hydroxybenzoic acid and phenol obtained after the experiment were characterized and confirmed with GC-MS analysis. The findings revealed the promising potentials of M. irregularis in PAH degradation and by extension green remediation technology. © 2020 The Authors

The indiscriminate disposal of olsalazine in the environment poses a threat to human health and natural ecosystems because of its cytotoxic and genotoxic nature. In the present study, degradation efficiency of olsalazine by the marine-derived fungus, Aspergillus aculeatus (MT492456) was investigated. Optimization of physicochemical parameters (pH. Temperature, Dry weight) and redox mediators {(2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), p-Coumaric acid and 1-hydroxybenzotriazole (HOBT)} was achieved with Response Surface Methodology (RSM)-Box-Behnken Design (BBD) resulting in 89.43% removal of olsalazine on 7th day. The second-order polynomial regression model was found to be statistically significant, adequate and fit with p

The impact of whole digestate (WD) and its fractions (solid [SD] and liquid [LD]) on 14C-phenanthrene mineralization in soil over 90 d contact time was investigated. The 14C-phenanthrene spiked soil was aged for 1, 30, 60 and 90 d. Analysis of water-soluble nitrogen, phosphorus, total (organic and inorganic) carbon, and quantitative bacterial count were conducted at each time point to assess their impact on mineralization of 14C-phenanthrene in soils. Indigenous catabolic activity (total extents, maximum rates and lag phases) of 14C-phenanthrene mineralization were measured using respirometric soil slurry assay. The soil amended with WD outperformed the SD and LD fractions as well as showed a shorter lag phase, higher rate and extent of mineralization throughout the study. The digestates improved (P < 0.05) the microbial population and nutritive content of the soil. However, findings showed that spiking soil with phenanthrene generally reduced the growth of microbial populations from 1 to 90 d and gave a lower nutritive content in comparison with the non-spiked soil. Also, soil fertility and bacteria count were major factors driving 14C-phenanthrene mineralization. Particularly, the non-phenanthrene degraders positively influenced the cumulative mineralization of 14C-phenanthrene after 60 d incubation. Therefore, the digestates (residue from anaerobic digestion) especially WD, which enhanced 14C-phenanthrene mineralization of the soil without minimal basal salts medium nor additional degraders should be further exploited for sustainable bioremediation of PAHs contaminated soil.

In this study, the thermal stability of a wood shaving biochar (WS, 650 °C), a chicken litter biochar (CL, 550 °C) and an activated carbon (AC, 1100 °C) were evaluated by combustion at 375 °C for 24 h to remove the labile non-carbonized organic matter. Results showed that WS and CL biochars were not thermally stable and can lose most of the organic C during combustion. The combusted WS and CL biochars retained considerable amounts of negative charge and displayed higher sorption for Cd (from 5.46 to 68.9 mg/g for WS and from 48.5 to 60.9 mg/g for CL). The AC retained 76.5% of its original C and became more negatively chargely after combustion, but its sorption for Cd slightly decreased (from 18.5 to 14.9 mg/g). This study indicated that after potential burning in wildfires (200–500 °C), biochars could have higher sorption capacity for metals by remaining minerals.

In this study, an acidic biochar and a neutral biochar were applied at 5 wt% into two soils for an 11-month incubation experiment. One Ferrosol soil (Ba) was slightly acidic with low organic matter and the other Dermosol soil (Mt) was slightly alkaline with high organic matter. The acidic (pH = 3.25) wood shaving (WS) biochar had no marked impact on nutrient levels, cation exchange capacity (CEC), pH and acid neutralization capacity (ANC) of either soil. By contrast, the neutral (pH = 7.00) chicken litter (CL) biochar significantly increased major soluble nutrients, pH, ANC of soil Ba. In terms of C storage, 87.9% and 69.5% WS biochar-C can be sequestrated as TOC by soil Ba and Mt, respectively, whereas only 24.0% of CL biochar-C stored in soil Ba and negligible amount in Mt as TOC. Biochars did not have significant effects on soil sorption capacity and sorption reversibility except that CL biochar increased sorption of soil Ba by around 25.4% and decreased desorption by around 50.0%. Overall, the studied acidic C rich WS biochar held little agricultural or remedial values but was favourable for C sequestration. The neutral mineral rich CL biochar may provide short-term agricultural benefit and certain sorption capacities of lower sorption capacity soils, but may be unlikely to result in heightened C sequestration in soils. This is the first study comprehensively examining functions of acidic and neutral biochars for their benefits as a soil amendment and suggests the importance of pre-testing biochars for target purposes prior to their large scale production. Refereed/Peer-reviewed

The Chinese Government is working to establish an effective framework in managing soil contamination. Heavy metal contamination is key to the discussion about soil quality, health and remediation in China. Soil heavy metal contamination in China is briefly reviewed and the concepts of background values and standards discussed. The importance of contaminated land and its management for China food security and urbanization are discussed. Priorities for China's next steps in developing an effective research and management regime are presented. We propose that critically important to the science-based risk assessment of contaminants in soils is the incorporation of speciation and bioavailability into the measurement and evaluation criteria. Consideration of soil biology/ecological endpoints will be necessary to protect ecosystem health. National and regional/local scenarios of land use type/usage will address residential/urban re-use of industrial land as well as varying agricultural scenarios.

In this study, the indigenous microbial mineralisation of 14C-phenanthrene in seven background soils (four from Norwegian woodland and three from the UK (two grasslands and one woodland)) was investigated. ∑PAHs ranged from 16.39 to 285.54 ng g−1 dw soil. Lag phases (time before 14C-phenanthrene mineralisation reached 5%) were longer in all of the Norwegian soils and correlated positively with TOC, but negatively with ∑PAHs and phenanthrene degraders for all soils. 14C-phenanthrene mineralisation in the soils varied due to physicochemical properties. The results show that indigenous microorganisms can adapt to 14C-phenanthrene mineralisation following diffuse PAH contamination. Considering the potential of soil as a secondary PAH source, these findings highlight the important role of indigenous microflora in the processing of PAHs in the environment.

Activated carbon amendment to contaminated soil has been proposed as an alternative remediation strategy to the management of persistent organic pollutant in soils and sediments. The impact of varying concentrations (0%, 0.01%, 0.1% and 1.0%) of different types of AC on the development of phenanthrene catabolism in soil was investigated. Mineralisation of (14)C-phenanthrene was measured using respirometric assays. The increase in concentration of CB4, AQ5000 or CP1 in soil led to an increase in the length of the lag phases. Statistical analyses showed that the addition of increasing concentrations of AC to the soil significantly reduced (P < 0.05) the extent of (14)C-phenanthrene mineralisation. For example, for CB4-, AQ5000- and CP1-amended soils, the overall extent of (14)C-phenanthrene mineralisation reduced from 43.1% to 3.28%, 36.9% to 0.81% and 39.6% to 0.96%, respectively, after 120 days incubation. This study shows that the properties of AC, such as surface area, pore volume and particle size, are important factors in controlling the kinetics of (14)C-phenanthrene mineralisation in soil.

When aromatic hydrocarbons are present in contaminated soils, they often occur in mixtures. The impact of four different (3-ring) nitrogen-containing polycyclic aromatic hydrocarbons (N-PAHs) on 12/14C-phenanthrene and 12/14C-benzo[a]pyrene (B[a]P) mineralisation in soil was investigated over a 90 d incubation period. The results revealed that both 12/14C-phenanthrene and 12/14C-benzo[a]pyrene showed no significant mineralisation in soils amended with 10 mg kg –1 and 100 mg kg –1 N-PAHs (p>0.05). However, increases in lag-phases and decreases in the rates and extents of mineralisation were observed, over time. Among the N-PAHs, benzo[h]quinoline impacted 14C-phenanthrene mineralisation with extended and diauxic lag phases. Furthermore,12/14C-B[a]P and 14C-benzo[a]pyrene–nitrogen-containing polycyclic aromatic hydrocarbons (14C-B[a]P-N-PAHs) amended soils showed extensive lag phases (> 21 d); with some 14C-B[a]P-N-PAH mineralisation recording

Lead (Pb) is a widespread heavy metal which is harmful to human health, especially to young children. To provide a human health risk assessment that is more relevant to real conditions, Pb bioavailability in soils is increasingly employed in the assessment procedure. Both in vivo and in vitro measurements for lead bioavailability are available. In vivo models are time- consuming and expensive, while in vitro models are rapid, economic, reproducible, and reliable while involving more uncertainties. Uncertainties in various measurements create difficulties in accurately predicting Pb bioavailability, resulting in the unnecessary remediation of sites. In this critical review, we utilised available data from in vivo and in vitro studies to identify the key parameters influencing the in vitro measurements, and presented uncertainties existing in Pb bioavailability measurements. Soil type, properties and metal content are reported to influence lead bioavailability; however, the differences in methods for assessing bioavailability and the differences in Pb source limit one’s ability to conduct statistical analyses on influences of soil factors on Pb bioavailability. The information provided in the review is fundamentally useful for the measurement of bioavailability and risk assessment practices.

Biochar is a carbon rich product from the incomplete combustion of biomass and it has been shown to reduce bioavailability of organic contaminants through adsorption. This study investigated the influence of 0%, 1%, 5% and 10% of two different particle sized wood biochars (≤2 mm and 3-7 mm) on the bioaccessibility of (14)C-phenanthrene (10 mg kg(-1)) in aged soil. The extent of (14)C-phenanthrene mineralisation by phenanthrene-degrading Pseudomonas sp. inoculum was monitored over a 14 day period in respirometric assays and compared to hydroxypropyl-β-cyclodextrin (HPCD) aqueous extraction. Notably, biochar amendments showed significant reduction in extents of mineralisation and HPCD extraction. Linear correlations between HPCD extractability and the total amount mineralised revealed good correlations, with 2 mm biochar showing a best fit (r(2) = 0.97, slope = 1.11, intercept = 1.72). Biochar reduced HPCD extractability and bioaccessibility of (14)C-phenanthrene to microorganisms in a similar manner. Biochar can aid risk reduction to phenanthrene exposure to biota in soil and HPCD can serve as a useful tool to assess the extent of exposure in biochar-amended soils.

Bioavailability has been used as a key indicator in chemical risk assessment yet poorly quantified risk factor. Worldwide, the framework used to assess potentially contaminated sites is similar, and the decisions are based on threshold contaminant concentration. The uncertainty in the definition and measurement of bioavailability had limited its application to environment risk assessment and remediation. Last ten years have seen major developments in bioavailability research and acceptance. The use of bioavailability in the decision making process as one of the key variables has led to a gradual shift towards a more sophisticated risk-based approach. Now a days, many decision makers and regulatory organisations 'more readily accept' this concept. Bioavailability should be the underlying basis for risk assessment and setting remediation goals of those contaminated sites that pose risk to environmental and human health. This paper summarises the potential application of contaminant bioavailability and bioaccessibility to the assessment of sites affected by different contaminants, and the potential for this to be the underlying basis for sustainable risk assessment and remediation in Europe, North America and Australia over the coming decade. Refereed/Peer-reviewed

In this study, the impact of different types of biochar and biochar ratios on the anaerobic digestion of citrus peel waste was investigated. Citrus peel has an inhibitory effect on anaerobic digestion. The presence of biochar had two effects: a reduction in the length of the lag phase and greater production of methane relative to citrus peel waste only incubations. The microbial lag phases decreased with increase in citrus peel to biochar ratios, with 2:1 having the longest lag phase of 9.4days and 1:3, the shortest, with the value of 7.5days. The cumulative methane production in incubations containing biochar and citrus peel ranged from 163.9 to 186.8ml CH4 gVS(-1), while citrus peel only produced 165.9ml CH4 gVS(-1). Examination of the biochar material revealed colonies of putative methanogens. The synergy of d-limonene adsorption and microbial immobilization by biochar appears to improve the performance of anaerobic digestion.

Soil organic matter is used to extrapolate the toxicity and bioavailability of organic pollutants between different soils. However, it has been shown that other factors such as microbial activity are crucial. The aim of this study was to investigate if sterilization can reduce differences in the fate and bioavailability of organic pollutants between different soils. Three natural soils with increasing total organic carbon (TOC) content were collected and three artificial soils were prepared to obtain similar TOCs. Soils were sterilized and spiked with C-14-pyrene and C-14-lindane. Total C-14 radioactivity, HPCD extractability, and bioaccumulation in Eisenia fetida were measured over 56 days. When compared to non-sterile soils, differences between the natural and artificial soils and the influence of soil-contaminant contact time were generally reduced in the sterile soils (especially with middle TOC). The results indicate the possibility of using sterile soils as "the worst case scenario" in soil ecotoxicity studies. (C) 2012 Elsevier Ltd. All rights reserved.

A strictly aerobic, halotolerant, Gram-stain-negative, rod-shaped bacterium, designated strain DG1253T, was isolated from a laboratory culture of the marine dinoflagellate Lingulodinium polyedrum (CCAP 1121/2). The strain was able to degrade two- and three-ring polycyclic aromatic hydrocarbons. It exhibited a narrow nutritional spectrum, preferring to utilize aliphatic and aromatic hydrocarbon compounds and small organic acids. Cells produced surface blebs and contained a single polar flagellum. The predominant isoprenoid quinone of strain DG1253T was Q-8. The fatty acid profile was dominated by C18 : 1ω7c. The mean DNA G+C content of strain DG1253T was 63.6±0.25 mol%. 16S rRNA gene sequence analysis placed this organism within the order Xanthomonadales of the class Gammaproteobacteria . Its closest relatives included representatives of the Hydrocarboniphaga – Nevskia – Sinobacter clade (≤89.9 % 16S rRNA gene sequence similarity) in the family Sinobacteraceae . On the basis of distinct phenotypic and genotypic characteristics, strain DG1253T is considered to represent a novel species of a new genus in the class Gammaproteobacteria , for which the name Algiphilus aromaticivorans gen. nov., sp. nov. is proposed. The type strain of the type species, Algiphilus aromaticivorans, is DG1253T ( = ATCC BAA-2243T = DSM 24793T). In addition, a new family, Algiphilaceae fam. nov., is proposed to accommodate the genus Algiphilus.

There is considerable current interest in using biochar (BC) as a soil amendment to sequester carbon to mitigate climate change. However, the implications of adding BC to agricultural soil for the environmental fate of pesticides remain unclear. In particular, the effect of biochars on desorption behavior of compounds is poorly understood. This study examined the influence of BC on pesticide chemical and biological accessibility using the herbicide isoproturon (IPU). Soils amended with 1% and 2% BC showed enhanced sorption, slower desorption, and reduced biodegradation of IPU. Addition of 0.1% BC had no effect on sorption, desorption or biodegradation of IPU. However, the mineralization of C-14-IPU was reduced by all BC concentrations, reducing by 13.6%, 40.1% and 49.8% at BC concentrations of 0.1%, 1% and 2% respectively. Further, the ratio of the toxic metabolite 4-isopropyl-aniline to intact IPU was substantially reduced by higher BC concentrations. Hydroxypropyl-beta-cyclodextrin (HPCD) extractions were used to estimate the IPU bioaccessibility in the BC-amended soil. Significant correlations were found between HPCD-extracted C-14-IPU and the IPU desorbed (%) (r(2) = 0.8518, p < 0.01), and also the C-14-IPU mineralized (%) (r(2) = 0.733; p < 0.01) for all BC-amended soils. This study clearly demonstrates how desorption in the presence of BC is intimately related to pesticide biodegradation by the indigenous soil microbiota. BC application to agricultural soils can affect the persistence of pesticides as well as the fate of their degradation products. This has important implications for the effectiveness of pesticides as well as the sequestration of contaminants in soils. (C) 2012 Elsevier Ltd. All rights reserved.

With increasing production of carbon nanoparticles (CNPs), environmental release of these entities becomes an ever-greater inevitability. However, many questions remain regarding their impact on soil microorganisms. This study examined the effects of long or short multiwalled carbon nanotubes (MWCNTs), C60 fullerene and fullerene soot in Gram-negative bacteria. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy was applied to derive signature spectral fingerprints of effects. A concentration-dependent response in spectral alterations was observed for each nanoparticle type. Long or short MWCNTs and fullerene soot gave rise to similar alterations to lipids, Amide II and DNA. The extent of alteration varies with nanoparticle size, with smaller short MWCNTs resulting in greater toxicity than long MWCNTs. Fullerene soot was the least toxic. C60 results in the most distinct and largest overall alterations, notably in extensive protein alteration. This work demonstrates a novel approach for assaying and discriminating the effects of CNPs in target systems.

The effect of cable oil concentration, nutrient amendment and bioaugmentation on cable oil component biodegradation in a pristine agricultural soil was investigated. Biodegradation potential was evaluated over 21 d by measuring cumulative CO(2) respiration on a Micro-Oxymax respirometer and (14)C-phenyldodecane mineralisation using a (14)C-respirometric assay. Cable oil concentration had a significant effect upon oil biodegradation. Microbial respiratory activity increased with increasing cable oil concentration, whereas (14)C-phenydodecane mineralisation decreased. Bioaugmentation achieved the best cable oil biodegradation performance, resulting in increases in cumulative CO(2) respiration, and maximum rates and extents of (14)C-phenyldodecane mineralisation. Generally, nutrient amendment also enhanced cable oil biodegradation, but not to the extent that degrader amendment did. Cable oil biodegradation was a function of (i) cable oil concentration and (ii) catabolic ability of microbial populations. Bioaugmentation may enhance cable oil biodegradation, and is dependent upon composition, cell number and application of catabolic inocula to soil.

Studies have shown that spinal cord stimulation (SCS) can reduce chronic pain by at least 50% over prolonged periods, improve function and quality-of-life, reduce requirements for healthcare resources and enable return to work in appropriately selected patients. However, SCS does not provide pain relief in all patients and is an expensive, labor intensive and invasive procedure with complications and ongoing management that requires specialists with specific skills and judgment. Multidisciplinary selection of appropriate patients for SCS is essential to achieve maximal benefit from the procedure. The aim of the article is to provide a clinical practice guide to the likely effectiveness of SCS in treating various types of chronic pain, as supported by the literature. The article will summarize indications and contraindications for SCS, provide guidance on the selection and timing for referral, and highlight the benefits and complications associated with the procedure.

The impact of fullerene soot (FS), single-walled (SWCNTs) and multi-walled (MWCNTs) carbon nano-tubes on the behaviour of two C-14-PAHs in sterile soil was investigated. Different concentrations of carbon nanomaterials (0, 0.05, 0.1 and 0.5%) were added to soil, and C-14-phenanthrene and C-14-benzo[a] pyrene extractability assessed over 80 d through dichloromethane (DCM) and hydroxypropyl-beta-cyclo-dextrin (HPCD) shake extractions. Total C-14-PAH activity in soils was determined by combustion, and mineralisation of C-14-phenanthrene was monitored over 14 d, using a catabolically active pseudomonad inoculum. No significant loss of C-14-PAH-associated activity from CNM-amended soils was observed over the 'aging' period. CNMs had a significant impact on HPCD-extractability of C-14-PANS: extractability decreased with increasing CNM concentration. Additionally, C-14-phenanthrene mineralisation was inhibited by the presence of CNMs at concentrations of >= 0.05%. Differences in overall extents of C-14-mineralisation were also apparent between CNM types. It is suggested the addition of CNMs to soil can reduce PAH extractability and bioaccessibility, with PAH sorption to CNMs influenced by CNM type and concentration. (C) 2010 Elsevier Ltd. All rights reserved.

Cypermethrin is a widely used insecticide that has caused concern due to its toxicity in the aquatic environment. As with all land applied pesticides, the most significant source of water pollution is from the soil, either due to leaching or washoff. The behaviour of cypermethrin in the soil controls the likelihood of future pollution incidents, with two of the most significant processes being the formation of bound residues and microbial degradation. The formation of bound residues and mineralisation was measured in four organically managed soils from the UK. The formation of bound residues was measured using three different extraction solutions, 0.01 M CaCl(2). 0.05 M HPCD and acetonitrile. Biodegradation was assessed by measurement of mineralisation of cypermethrin to CO(2). The formation of bound residues varied according to extraction method, soil type and length of ageing. In two of the four soils studied, acetonitrile extractability decreased from 100% initially to 12-14% following 100 d ageing. The extent of mineralisation increased after 10-21 d ageing, reaching 33% of remaining activity in one soil, however following 100 d ageing the extent of mineralisation was significantly reduced in three out of the four soils. As with the formation of bound residues, mineralisation was impacted by soil type and length of ageing. (C) 2010 Elsevier Ltd. All rights reserved.

Slurry-based mineralisation assays are widely used to investigate contaminant biodegradation in soil; however, the importance of shaking speed on microbial degradation has not been considered. This study investigated the mineralisation of (14)C-analogues of phenanthrene, hexadecane and octacosane, shaken at 0, 25 and 100 rpm. The results showed that the fastest rates and highest levels of mineralisation in 0 d aged soils were in the highly agitated conditions (100 rpm). However, the highest levels of mineralisation in 500 d aged soil were found in the gently shaken conditions (25 rpm), with the levels of mineralisation significantly (p

Despite numerous reviews suggesting that microbial biosensors could be used in many environmental applications, in reality they have failed to be used for which they were designed. In part this is because most of these sensors perform in an aqueous phase and a buffered medium, which is in contrast to the nature of genuine environmental systems. In this study, a range of non-exhaustive extraction techniques (NEETs) were assessed for (i) compatibility with a naphthalene responsive biosensor and (ii) correlation with naphthalene biodegradation. The NEETs removed a portion of the total soil naphthalene in the order of methanol > HPCD > βCD > water. To place the biosensor performance to NEETs in context, a biodegradation experiment was carried out using historically contaminated soils. By coupling the HPCD extraction with the biosensor, it was possible to assess the fraction of the naphthalene capable of undergoing microbial degradation in soil.

The characteristics of natural soils often vary from those of artificial soil (e.g. OECD), which may lead to substantial differences in the bioavailability of test substances. The aim of this investigation was to characterise the development of phenanthrene catabolism in both natural and artificial soils with varying total organic carbon (TOC) content after 1, 14, 42 and 84 d soil-phenanthrene contact time. Indigenous catabolic activity was measured via the addition of 14C-phenanthrene using the respirometric soil slurry assay. Notably, the lag phases, fastest rates and total extents of 14C-phenanthrene degradation were relatively comparable in soils with similar TOC content after 1 d contact time. However, natural soils generally exhibited significantly shorter lag phases, faster rates and higher extents of mineralisation, than their artificial counterparts after 42 and 84 d contact time. Such findings suggest that the extrapolation of results from artificial soils to real/natural soils may not be straightforward.

Aliphatic hydrocarbons make up a substantial portion of organic contamination in the terrestrial environment. However, most studies have focussed on the fate and behaviour of aromatic contaminants in soil. Despite structural differences between aromatic and aliphatic hydrocarbons, both classes of contaminants are subject to physicochemical processes, which can affect the degree of loss, sequestration and interaction with soil microflora. Given the nature of hydrocarbon contamination of soils and the importance of bioremediation strategies, understanding the fate and behaviour of aliphatic hydrocarbons is imperative, particularly microbe-contaminant interactions. Biodegradation by microbes is the key removal process of hydrocarbons in soils, which is controlled by hydrocarbon physicochemistry, environmental conditions, bioavailability and the presence of catabolically active microbes. Therefore, the aims of this review are (i) to consider the physicochemical properties of aliphatic hydrocarbons and highlight mechanisms controlling their fate and behaviour in soil; (ii) to discuss the bioavailability and bioaccessibility of aliphatic hydrocarbons in soil, with particular attention being paid to biodegradation, and (iii) to briefly consider bioremediation techniques that may be applied to remove aliphatic hydrocarbons from soil.

This study investigates the impact of different types of carbon nanomaterials (CNMs) namely C60, multi-walled carbon nanotubes (MWCNTs) and fullerene soot on the catabolism of (14)C-phenanthrene in soil by indigenous microorganisms. Different concentrations (0%, 0.01%, 0.1% and 1%) of the different CNMs were blended with soil spiked with 50 mg kg(-1) of (12)C-phenanthrene, and aged for 1, 25, 50 and 100 days. An increase in the concentration of MWCNT- and FS-amended soils showed a significant difference (P = 0.014) in the lag phase, maximum rates and overall extent of (14)C-phenanthrene mineralisation. Microbial cell numbers did not show an obvious trend, but it was observed that control soils had the highest population of heterotrophic and phenanthrene degrading bacteria at all time points.

In this study, an aqueous-based hydroxypropyl-beta-cyclodextrin (HPCD) extraction technique was assessed for its capacity to determine the microbially degradable fraction of mono- and polycyclic aromatic hydrocarbons in four dissimilar soils. A linear relationship (slope = 0.90; R-2 = 0.89), approaching 1:1 between predicted and observed phenanthrene mineralization, was demonstrated for the cyclodextrin extraction; however, the water only extraction underestimated the microbially available fraction by a factor of three (slope = 3.35; R-2 = 0.64). With respect to determining the mineralizable fraction of p-cresol in soils, the cyclodextrin extraction (slope = 0.94; R-2 = 0.84) was more appropriate than the water extraction (slope = 1.50; R-2 = 0.36). Collectively, these results suggested that the cyclodextrin extraction technique was suitable for the prediction of the mineralizable fraction of representative PAHs and phenols present in dissimilar soils following increasing soil-contaminant contact times. The assessment of the microbial availability of contaminants in soils is important for a more representative evaluation of soil contamination. (c) 2006 Elsevier Ltd. All rights reserved.

The aim of this study was to characterize the behaviour of a PAH-degrading bacterium to determine whether mineralization plateaus as a result of substrate removal, a decrease in microbial activity or nutrient availability in sterile soils over time. To investigate this, the mineralization of 14C-phenanthrene was measured until it plateaued; subsequently, additional 14C-phenanthrene, catabolic inocula or nutrients were introduced and mineralization was measured for a further 10 d. Cell numbers were also measured together with 14C-uptake into microbial biomass. Freshly added 14C-phenanthrene was rapidly metabolised by the microorganisms. Neither the addition of a catabolic inoculum nor nutrients affected the extent of 14C-phenanthrene mineralization. Cell numbers remained constant over time, with only a small amount of the 14C-activity incorporated into the microbial biomass. This study indicated that the termination of mineralization was due to the removal of available phenanthrene and not decreasing cellular activity or cell death. The mineralization values also correlated with 14C-phenanthrene extractability using β-cyclodextrin. Mineralization can estimate the microbial availability of 14C-contaminants in soil.

The influence of pyrene added in a single application (0, 50, 100 and 200 mg kg(-1)) was investigated in Multiple applications (1 X 50, 2 X 50 and 4 X 50 mg kg(-1)) on the evolution of catabolic activity in a pristine pasture soil. The microbial community's ability to degrade pyrene was assessed at 0, 4, 8 and 12 weeks by the mineralization of added C-14-pyrene. Significant mineralization (> 5%) of added C-14-pyrene only occurred after 4 weeks soil-pyrene contact time in most of the pyrene-amended soils. Pyrene-amended soils showed statistically significantly shorter (P < 0.05) lag times compared to the control soil after 8 and 12 weeks soil-pyrene contact time. Further, the rates of degradation increased in the presence of pyrene, peaking at 8 weeks. In terms of the overall extents of pyrene mineralization, there were statistically significant increases (P < 0.05) between 4 and 8 weeks, with little difference between 8 and 12 weeks, with the general trend that an increase in pyrene concentration resulted in higher levels of mineralization. Increasing the concentration and number of pyrene additions can have a significant impact on the adaptation of the soil microflora to degrade pyrene over time. (c) 2005 Elsevier Ltd. All rights reserved.

There is a need to develop risk biomarkers during the remediation of contaminated land. We employed the earthworm, Aporrectodea longa (Ude), to determine whether genotoxicity measures could be applied to this organism's intestinal tissues. Earthworms were added, for 24h or 7 days, to soil samples spiked with benzo[a]pyrene (B[a]P) and/or lindane. After exposure, intestinal tissues (crop/gizzard or intestine) were removed prior to the measurement in disaggregated cells of DNA single-strand breaks (SSBs) by the alkaline comet assay. Damage was quantified by comet tail length (CTL, microm). B[a]P 24-h exposure induced dose-related increases (P0.0001) in SSBs. Earthworm intestine was significantly (P0.0001) more susceptible than crop/gizzard to B[a]P and/or lindane. However, both tissues appeared to acquire resistance following 7-day exposure. B[a]P-DNA adducts, measured by (32)P-postlabelling, showed a two-adduct-spot pattern. This preliminary investigation suggests that earthworm tissues may be incorporated into genotoxicity assays to facilitate hazard identification within terrestrial ecosystems.

The role of native soil microorganisms in the formation and release of non-extractable (14)C-residues, previously treated with (14)C-Dicamba, was investigated to examine their significance to the longer-term environmental effects on non-extractable pesticide residues. A 90 d study compared the fate of Dicamba under sterile and non-sterile regimes. In addition, soils were aged for 30 d and repeatedly extracted with a 0.01 M CaCl(2) solution, to an extraction end point, to produce non-extractable residues. The extracted soil containing non-extractable residues was mixed with clean soil that had been freshly spiked with non-labeled Dicamba at 0.2 mg kg(-1) to increase the bulk volume of the soil and stimulate microbial activity. Sub-samples were then introduced into microcosms to compare the extent of microbially facilitated release and mineralisation with release rates in sterile microcosms. The results show that microorganisms play a significant role in the formation and release of non-extractable Dicamba residues. The release of (14)C-activity in sterile microcosms was linked to physical mixing of the extracted soil with field soil prior to the beginning of the incubations. The released (14)C-activity may be further mineralized, reincorporated into humus, or taken up by plants or other soil inhabiting biota.

Laboratory based studies on the fate of organic contaminants in soil typically requires the test compound(s) to be spiked into the test medium. Consequently, such studies are inherently dependent on the homogeneity of the contaminant within the spiked soil. Three blending methods were compared for the addition of a phenanthrene-transformer oil mixture into field-wet soil. Spiking homogeneity, reproducibility and artefacts were assessed based on dichloromethane and hydroxypropyl-beta-cyclodextrin chemical extractability, and bacterial mineralization. Spiking using a stainless-steel spoon, consistently produced good spike homogeneity as determined by sample oxidation, chemical extraction and mineralization, and was consistently more reliable than either the Waring blender or modified bench drill. Overall, neither transformer oil-concentration nor blending method influenced chemical extractability or mineralization of the PAH following 1 day equilibration. In general, spiking procedures require validation prior to use, as homogeneity cannot be assured.

Chemical pollution of the environment has become a major source of concern. In particular, many studies have investigated the impact of pollution on biota in the environment. Studies on metalliferous contaminated mine spoil wastes have shown that some soil organisms have the capability to become resistant to metal/metalloid toxicity. Earthworms are known to inhabit arsenic-rich metalliferous soils and, due to their intimate contact with the soil, in both the solid and aqueous phases, are likely to accumulate contaminants present in mine spoil. Earthworms that inhabit metalliferous contaminated soils must have developed mechanisms of resistance to the toxins found in these soils. The mechanisms of resistance are not fully understood; they may involve physiological adaptation (acclimation) or be genetic. This review discusses the relationships between earthworms and arsenic-rich mine spoil wastes, looking critically at resistance and possible mechanisms of resistance, in relation to soil edaphic factors and possible trophic transfer routes.

This paper describes the induction of phenanthrene-catabolism within Phase II mushroom compost resulting from its incubation with (1) phenanthrene, and (2) PAH-contaminated soil. Respirometers measuring mineralization of freshly added 14C-9-phenanthere were used to evaluate induction of phenanthrene-catabolism. Where pure phenanthrene (spiked at a concentration of 400 mg kg(-1) wet wt.) was used to induce phenanthrene-catabolism in compost, induction was measurable, with maximal mineralization observed after 7 weeks phenanthrene-compost contact time. Where PAH-contaminated soil was used to induce phenanthrene-catabolism in un-induced compost, induction was observed after 5 weeks soil-compost contact time. Microcosm-scale amelioration of soil contaminated with 14C-phenanthrene (aged in soil for 516 days prior to incubation with compost) indicated that both induced (using pure phenanthrene) and uninduced Phase II mushroom composts were equally able to promote degradation of this soil-associated contaminant. After 111 days incubation time, 42.7 +/- 6.3% loss of soil-associated phenanthrene was observed in the induced-compost soil mixture, while 36.7 +/- 2.9% loss of soil-associated phenanthrene was observed in the uninduced-compost soil mixture. These results are notable as they indicate that while pre-induction of phenanthrene-catabolism within compost is possible, it does not significantly increase the extent of degradation when the compost is used to ameliorate phenanthrene-contaminated soil. Thus, compost could be used directly in the amelioration of contaminated land i.e. without pre-induction of catabolism.

Highlights • We studied the resistance and resilience of soil microbial communities. • There was a significant concentration-dependent impact on dehydrogenase activity. • Significant impacts on nematode and fungal communities were also observed., The application of plant protection products has the potential to significantly affect soil microbial community structure and function. However, the extent to which soil microbial communities from different trophic levels exhibit resistance and resilience to such compounds remains poorly understood. The resistance and resilience responses of a range of microbial communities (bacteria, fungi, archaea, pseudomonads, and nematodes) to different concentrations of the strobilurin fungicide, azoxystrobin were studied. A significant concentration-dependent decrease, and subsequent recovery in soil dehydrogenase activity was recorded, but no significant impact on total microbial biomass was observed. Impacts on specific microbial communities were studied using small subunit (SSU) rRNA terminal restriction fragment length polymorphism (T-RFLP) profiling using soil DNA and RNA. The application of azoxystrobin significantly affected fungal and nematode community structure and diversity but had no impact on other communities. Community impacts were more pronounced in the RNA-derived T-RFLP profiles than in the DNA-derived profiles. qPCR confirmed that azoxystrobin application significantly reduced fungal, but not bacterial, SSU rRNA gene copy number. Azoxystrobin application reduced the prevalence of ascomycete fungi, but increased the relative abundance of zygomycetes. Azoxystrobin amendment also reduced the relative abundance of nematodes in the order Enoplia, but stimulated a large increase in the relative abundance of nematodes from the order Araeolaimida.

Carbon nanotubes (CNTs) are considered promising materials in nanotechnology. We quantified CNT accumulation by the alga Desmodesmus subspicatus. Cells were exposed to radiolabeled CNTs ((14)C-CNTs; 1 mg/L) to determine uptake and association, as well as elimination and dissociation in clear media. Attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) was used to detect effects of CNTs on algae. CNT-cell interactions were visualized by electron microscopy and related to alterations in their cell composition. A concentration factor of 5000 L/kg dry weight was calculated. Most of the material agglomerated around the cells, but single tubes were detected in the cytoplasm. Computational analyses of the ATR-FTIR data showed that CNT treated algae differed from controls at all sampling times. CNT exposure changed the biochemical composition of cells. The fact that CNTs are bioavailable for algae and that they influence the cell composition is important with regard to environmental risk assessment of this nanomaterial.

A method was developed to determine trace concentrations of a range of individual PCB congeners in biological samples (serum, food and faeces) using GC-MS, to prepare a mass balance of PCBs in humans. A simple method for the analysis of PCBs in human serum, which excluded an extraction step, was first employed. Results indicated that the recoveries of 13C12 PCB spikes were variable. A soxhlet extraction step was added and was found to be efficient and reproducible. A quality control routine and method validation results are presented. In batch tests of the methods presented it was found that the serum analysis method gave within batch mean 13C12 spike recoveries of 98 – 120% and standard deviations between 6 and 20%. The food/faeces analysis method gave within-batch mean 13C12 spike recoveries of 88 – 100%, and within batch standard deviations between 4 and 12%. The batch to batch mean recovery for serum analysis was 100%, with an RSD of 9% for high spikes and 10% for low spikes. For food/faeces analysis the batch to batch average recovery was 110%, with an RSD of 5% for high spikes and 9% for low spikes.

Because phenols are one of the most common groups of organic pollutants in the aquatic environment, heterotrophic growth-linked biodegradation of phenol and its methylated homologues by the eukaryotic alga Ochromonas danica (CCAP 933/2B) was investigated. The alga grew heterotrophically on phenol and mixtures of phenol with o- or p-cresols, or with 2,5-, 2,6-, 3,4- or 3,5-xylenols as the sole sources of carbon in the dark at 25 degrees C. Commensurate with growth, the alga removed phenol, both cresol isomers and 2,5- and 3,4-xylenols from the growth media over the incubation periods. In every case, phenol was removed preferentially to the methylated cosubstrates, but the rates of removal for phenol were slower than in incubations where phenol was the sole carbon source.

Composting is an aerobic process which allows the rapid proliferation of a variety of microbial groups such as aerobic actinomycetes, bacilli and fungi. Waste mushroom compost is readily available as a byproduct, with 400,000-500,000 tons produced per year in the UK, and is capable of acting as a carrier for the xenobiotic-degrading microorganisms and as a nutrient source in contaminated soils. Bacteria, particularly actinomycetes, were targeted as potential PCP degraders because of their ubiquitous distribution in composts, their ability to colonize composts by mycelial growth and to degrade a wide range of aromatic compounds using either endoor extracellular enzymes ( McCarthy and Williams, 1992 ). Semple and Fermor (1995) showed that actinomycetes, isolated from PCP-contaminated mushroom compost, were capable of attacking this highly chlorinated phenol. However, the individual putative PCP-degrading isolates tested did not achieve appreciable mineralization of UL-14C-PCP. It may be that a diverse population working in concert is required to biodegrade the chlorophenol to CO2 and H2O. However, biotransformation of PCP is a possibility given that the Saccharomonaspora viridis strains, Streptomyces spp., and Thermomonospora spp. appear to remove the PCP from the growth media ( Semple and Fermor, 1995 ). Mineralization of PCP by composts was found to be more impressive compared to that of the actinomycete isolates. This suggests that the physical relationship between compost solid substrate and the populating microflora play an important role in the degradation of PCP. A key phenomenon to be considered when applying a bioremediative technology is chemical ageing and its relationship to bioavailability of the contaminant to the soil microflora ( Hatzinger and Alexander, 1995 ). The longer an organic compound is in a given soil environment, the more likely it is to be affected by chemical and physical processes, such as diffusion into soil micropores, partitioning into soil organic matter, strong surface adsorption and covalent binding to soil constituents. These processes result in a decrease in the bioavailability of the compound to the degradative microorganisms, thus reducing the effectiveness of the treatment. However, it has been suggested that desorption of compounds (previously unavailable), leading to an increase in bioavailability, may occur at elevated temperatures typically generated under composting conditions ( Pignatello and Xing, 1996 ). This will require further investigation to understand the processes involved and to characterize its effects on the degradation of PCP by mushroom composts.

The first strobilurin fungicides were introduced in 1996, and have since been used in a vast array of disease/plant systems worldwide. The strobilurins now consist of 16 compounds and represent the 2nd most important fungicide group worldwide with 15% of the total fungicide market share. Strobilurins are moderately persistent in soil, and some degradation products (e.g. azoxystrobin acid) have been detected as contaminants of freshwater systems. Little is currently known about the transformation processes involved in the biodegradation of strobilurins or the microbial groups involved. Using sequential soil and liquid culture enrichments, we isolated two bacterial strains which were able to degrade the most widely used strobilurin, azoxystrobin, when supplied as a sole carbon source. 16S rRNA showed that the strains showed homology to Cupriavidus sp. and Rhodanobacter sp. Both isolated strains were also able to degrade the related strobilurin compounds trifloxystrobin, pyraclostrobin, and kresoxim-methyl. An additional nitrogen source was required for degradation to occur, but the addition of a further carbon source reduced compound degradation by approximately 50%. However, (14)C radiometric analysis showed that full mineralisation of azosxystrobin to (14)CO2 was negligible for both isolates. 16S rRNA T-RFLP analysis using both DNA and RNA extracts showed that degradation of azoxystrobin in soil was associated with shifts in bacterial community structure. However, the phylotypes which proliferated during degradation could not be attributed to the isolated degraders.

Mixtures of polycyclic aromatic hydrocarbons (PAHs) and heavy metals are of major concern in contaminated soil. Biodegradation of PAHs in metal-contaminated soils is complicated because metals are toxic and cannot be degraded by biological processes. This investigation considered the effects of Zn and Cu (50, 100, 500 and 1,000 mg/kg) on 14C-phenanthrene biodegradation in soil over 60-day contact time. The presence of Zn at all concentrations and low concentrations of Cu (50 and 100 mg/kg) had no significant effect (p > 0.05) on the development of phenanthrene catabolism; however, at higher Cu concentrations, the development of phenanthrene catabolism and bacterial cell numbers were significantly reduced (p

The work presented in this paper investigated the effects of plant species composition, species diversity and soil fertility on biodegradation of (14)C-phenanthrene in soil. The two soils used were of contrasting fertility, taken from long term unfertilised and fertilised grassland, showing differences in total nitrogen content (%N). Plant communities consisted of six different plant species: two grasses, two forbs, and two legume species, and ranged in species richness from 1 to 6. The degradation of (14)C-phenanthrene was evaluated by measuring indigenous catabolic activity following the addition of the contaminant to soil using respirometry. Soil fertility was a driving factor in all aspects of (14)C-phenanthrene degradation; lag phase, maximum rates and total extents of (14)C-phenanthrene mineralisation were higher in improved soils compared to unimproved soils. Plant identity had a significant effect on the lag phase and extents of mineralisation. Soil fertility was the major influence also on abundance of microbial communities.

The rate of pesticide biodegradation does not remain constant with time, and is dependent on the physico-chemical properties of the soil and of the pesticide as well as on the biology of the soil. Prolonged or repeated contact between soil microbes and pesticides has been shown to result in an increase in the rate and extent of biodegradation. This work assessed the impact of the soil:water ratio on measurement of catabolic induction for ¹⁴C-isoproturon, ¹⁴C-diazinon and ¹⁴C-cypermethrin. Slurrying (1:1 and 1:3 soil:water) with agitation resulted in significantly higher rates and extents of mineralisation than the non-slurried system (P ≤ 0.05; 1:0 soil:water), except for the mineralisation of ¹⁴C-diazinon where the greatest extent of mineralisation occurred in non-slurried soil. Slurrying without agitation resulted in the significant lower mineralisation in all cases (P ≤ 0.05). There was a significant interaction between the soil:water ratio and length of contact (P ≤ 0.05). Whilst the use of slurried systems can enhance the extent and rate of mineralisation, there is no improvement in reproducibility, and so for the measurement of catabolic induction, the use of field conditions will lead to a more environmentally relevant measurement.

The behaviour of diazinon in the soil determines the likelihood of further pollution incidents, particularly leaching to water. The most significant processes in the control of the fate of diazinon in the soil are microbial degradation and the formation of bound residues. Soils from four sites in the UK were amended with diazinon and its 14C labelled analogue and incubated for 100 days. After 0, 10, 21, 50 and 100 days, the formation of bound residues was assessed by solvent extraction, and the microbial degradation of diazinon by mineralisation assay. In microbially active soils, diazinon is degraded rapidly, reducing the risk of future pollution incidents. However, where there was limited mineralisation there was also significantly lower formation of bound residues, which may lead to water pollution via leaching. The formation of bound residues was dependent on extraction type. Acetonitrile extraction identified bound residues in all soils, with the bound residue fraction increasing with increasing incubation time.

A dynamic multimedia fugacity model was used to evaluate the partitioning and fate of petroleum hydrocarbon fractions and aromatic indicator compounds within the soil oil matrix of three biopiles Each biopile was characterised by four compartments air, water, soil solids and non-aqueous phase liquid (NAPL) Equilibrium partitioning in biopile A and B suggested that most fractions resided in the NAPL, with the exception of the aromatic fraction with an equivalent carbon number from 5 to 7 (EC5-7) In Blopile C, which had the highest soil organic carbon content (13%), the soil solids were the most important compartment for both light aliphatic fractions (EC5-6 and EC6-8) and aromatic fractions, excluding the EC16-21 and EC21-35 Our starting hypothesis was that hydrocarbons do not degrade within the NAPL This was supported by the agreement between predicted and measured hydrocarbon concentrations in Biopile B when the degradation rate constant in NAPL was set to zero In all scenarios, biodegradation in soil was predicted as the dominant removal process for all fractions, except for the aliphatic EC5-6 which was predominantly lost via volatilization. The absence of an explicit NAIL phase in the model yielded a similar prediction of total petroleum hydrocarbon (TPH) behaviour, however the predicted concentrations in the air and water phases were significantly increased with consequent changes in potential mobility Further comparisons between predictions and measured data, particularly concentrations in the soil mobile phases, are required to ascertain the true value of including an explicit NAPL in models of this kind (C) 2010 Elsevier Ltd All rights reserved

A six month field scale study was carried out to compare windrow turning and biopile techniques for the remediation of soil contaminated with bunker C fuel oil. End-point clean-up targets were defined by human risk assessment and ecotoxicological hazard assessment approaches. Replicate windrows and biopiles were amended with either nutrients and inocula, nutrients alone or no amendment. In addition to fractionated hydrocarbon analysis, culturable microbial characterisation and soil ecotoxicological assays were performed. This particular soil, heavy in texture and historically contaminated with bunker fuel was more effectively remediated by windrowing, but coarser textures may be more amendable to biopiling. This trial reveals the benefit of developing risk and hazard based approaches in defining end-point bioremediation of heavy hydrocarbons when engineered biopile or windrow are proposed as treatment option. (c) 2010 Elsevier Ltd. All rights reserved.

The influence of PAH chemical structure and concentration, added in either single (75 or 300 mg kg−1) or multiple (2 × 75, 2 × 150 or 4 × 75 mg kg−1) applications as single- or multiple-contaminant systems, on the development of PAH biodegradation in a pristine soil was investigated. Development in microbial catabolic ability was assessed at 0, 28, 56 and 84 d by monitoring 14C-naphthalene, 14C-phenanthrene and 14C-pyrene mineralisation over 14 d in respirometric assays. The presence of other contaminants influenced the ability of the indigenous microflora to mineralise structurally different contaminants over time. 14C-Naphthalene mineralisation was inhibited by the presence of other contaminants; whereas the presence of naphthalene significantly enhanced rates of mineralisation in multiple-contaminant systems containing 14C-phenanthrene and 14C-pyrene. Generally, increasing the number of contaminant applications has implications for catabolic activity of soil microbes. It is suggested the toxic nature of PAHs retarded mineralisation at increased contaminant concentrations. The simultaneous effects of PAH concentration, contaminant mixture and repeated application on the development of catabolic activity in soil.