Human health‐based soil cleanup guidelines for diesel fuel no. 2

Soil cleanup guidelines were developed for diesel fuel No. 2 that are protective of human health. Guidelines were conservatively based on a residential land use scenario. This scenario estimates human health risks associated with long‐term exposure to site soil via the inhalation, dermal, and ingestion routes of exposure. Lifetime dermal cancer studies were selected as the basis for deriving a safe level of diesel fuel in soil. Soil cleanup guidelines for diesel fuel No. 2 ranged from 1166 to 11,287 mg/kg for adult or child residents and represent contaminant levels that pose acceptable health risks for both present and proposed future uses of a site.     

Anaerobic biodegradation of diesel fuel-contaminated wastewater in a fluidized bed reactor

Diesel fuel spills have a major impact on the quality of groundwater. In this work, the performance of an Anaerobic Fluidized Bed Reactor (AFBR) treating synthetic wastewater is experimentally evaluated. The wastewater comprises tap water containing 100, 200 and 300 mg/L of diesel fuel and nutrients. Granular, inert, activated carbon particles are employed to provide support for biomass inside the reactor where diesel fuel is the sole source of carbon for anaerobic microorganisms. For different rates of organic loading, the AFBR performance is evaluated in terms of the removal of diesel fuel as well as chemical oxygen demand (COD) from wastewater. For the aforementioned diesel fuel concentrations and a wastewater flow rate of 1,200 L/day, the COD removal ranges between 61.9 and 84.1%. The concentration of diesel fuel in the effluent is less than 50 mg/L, and meets the Level II groundwater standards of the MUST guidelines of Alberta.     

The effects of carbon sources and micronutrients in fermented whey on the biodegradation of n-hexadecane in diesel fuel contaminated soil

Fermented whey has previously been shown to stimulate biodegradation of n-hexadecane in diesel contaminated soils. The proposed explanation for the stimulatory effect is that fermented whey provides easily accessible carbon and micronutrients, which give rise to an increased degrading biomass.The objective of this work has been to investigate the role of the different carbon sources and vitamins in fermented whey on the microbial degradation of n-hexadecane in soil.The effects of lactose, lactate, vitamins and free amino acids were tested in combinations according to a full factorial design experiment, at concentrations corresponding to those present in fermented whey. The target substance was ¹⁴C-labeled n-hexadecane in nutrient amended soil microcosms contaminated with 5000 mg diesel fuel kg⁻¹ dw. Biodegradation was monitored by determination of evolved ¹⁴CO₂.Significant effects on the biodegradation of n-hexadecane were observed for lactate and amino acids additions in a sandy soil. Lactate showed both an inhibitory effect in the early phase of the experiment and a stimulatory effect in the later phase. The effect of amino acids was slightly stimulatory, mainly evident as a shortening of the lag time.The degree of n-hexadecane degradation at the end of the experiment was correlated with the total concentration of organic compounds added to the soil.Scientific relevanceThere are a handful papers describing the potential of using organic amendments (often industrial by-products) with a content of both easily accessible carbon and micronutrients, to enhance the bioremediation of polluted soils. Enhanced biodegradation is often reported and the proposed explanations are that the combination of easily accessible carbon and micronutrients increases the degrading biomass.In this paper, we examine the effect of fermented whey on the degradation of n-hexadecane and correlate the observed effects on the biodegradation with the main components lactate, amino acids, lactose and B-vitamins. This has to our knowledge never been done before.     

Versatility of soil column experiments to study biodegradation of halogenated compounds under environmental conditions

Soil column experiments were performed to obtain insight in the different biological and physico-chemical processes affecting biodegradation of halogenated compounds under natural conditions in a water infiltration site. Lower chlorinated aromatic compounds could be degraded under aerobic conditions, whereas highly chlorinated compounds and chlorinated aliphatic compounds were mainly transformed under anaerobic conditions. Microorganisms which derive energy from reductive dechlorination were enriched and characterized. It was found that microbes could adapt to using chlorinated benzenes by evolution of new enzyme specificities and by exchange of genetic material. For halogenated pollutants, which are generally hydrophobic, sorption processes control the concentration available for biodegradation. The effects of very low concentrations of halogenated compounds on their biodegradability are described. The use of isolated bacterial strains to enhance biodegradation was evaluated with respect to their temperature-related activity and to their adhesion properties.Abbreviations 3-CB 3-chlorobenzoate - DCB dichlorobenzene - HCH hexachlorocyclohexane - IS insertion sequence - PER tetrachloroethylene - S_min minimal substrate concentration for growth - TCB trichlorobenzene - TRI trichloroethylene - filtration coefficient     

Anaerobic degradation of No. 2 diesel fuel in the wetland sediments of Barataria-Terrebonne estuary under various electron acceptor conditions

The biodegradation of No. 2 diesel fuel under anaerobic conditions was investigated using sediments collected from wetlands of Barataria-Terrebonne estuary in Louisiana. The results indicated enhanced biodegradation of diesel fuel under sulfate-reducing, nitrate-reducing, methanogenic, and mixed electron acceptor conditions. However, the rate of diesel degradation was the highest under mixed electron acceptor conditions followed in order by sulfate-reducing, methanogenic, and nitrate-reducing conditions. Under mixed electron acceptor condition, 99% removal of diesel fuel was achieved within 510 days, while under sulfate-reducing condition 62% degradation of diesel fuel was observed for the same period. Diesel fuel was also degraded to a smaller extent in the culture condition where electron acceptors were not supplemented (natural attenuation condition). This study showed evidence for enhanced diesel fuel metabolism in a mixed microbial population system similar to any contaminated field site, where a heterogeneous microbial population exists.     

Phytoremediation of subarctic soil contaminated with diesel fuel

The effects of several plant species, native to northern latitudes, and different soil amendments, on diesel fuel removal from soil were studied. Plant treatments included Scots Pine (Pinus sylvestris), Poplar (Populus deltoides x Wettsteinii), a grass mixture (Red fescue, Fesuca rubra; Smooth meadowgrass, Poa pratensis and Perennial ryegrass, Lolium perenne) and a legume mixture (White clover, Trifolium repens and Pea, Pisum sativum). Soil amendments included NPK fertiliser, a compost extract and a microbial enrichment culture. Diesel fuel disappeared more rapidly in the legume treatment than in other plant treatments. The presence of poplar and pine enhanced removal of diesel fuel, but removal under grass was similar to that with no vegetation. Soil amendments did not enhance diesel fuel removal significantly. Grass roots accumulated diesel-range compounds. This study showed that utilisation of selected plants accelerates removal of diesel fuel in soil and may serve as a viable, low-cost remedial technology for diesel-contaminated soils in subarctic regions.     

Anaerobic biodegradation of polychlorinated biphenyls by a microbial consortium originated from uncontaminated paddy soil

Polychlorinated biphenyls (PCBs) in Kanechlor-300 and -400 mixtures dissipated significantly compared with a sterilized control under anaerobic conditions in three Japanese paddy soils with no history of PCB contamination, demonstrating the anaerobic microbial degradation of PCBs. The PCB-degrading activity was maintained successfully in a static flooded soil medium for more than 3 years by serial transfer at intervals of 56 days (13 transfers). Ortho-, meta-, and para-substituted PCBs, 15.2 ± 9.9 mol% in total, were significantly degraded after 56 days of incubation. Analysis of menaquinones-6 and -7 and cloning of 16S rRNA gene fragments from a polymerase chain reaction denaturing gradient gel electrophoresis (DGGE) profile indicated the predominance of Firmicutes in the consortium. A PCR-based identification of the gene fragments showed the frequent presence of Desulfitobacterium sp., but not Dehalobacter sp. or Dehalococcoides sp., in the consortium. It is proposed that Japanese paddy soils with no history of PCB contamination contain an anaerobic microbial consortium consisting predominantly of Firmicutes that have the potential for anaerobic degradation of PCB.     

The effects of carbon sources and micronutrients in fermented whey on the biodegradation of n-hexadecane in diesel fuel contaminated soil

Fermented whey has previously been shown to stimulate biodegradation of n-hexadecane in diesel contaminated soils. The proposed explanation for the stimulatory effect is that fermented whey provides easily accessible carbon and micronutrients, which give rise to an increased degrading biomass.The objective of this work has been to investigate the role of the different carbon sources and vitamins in fermented whey on the microbial degradation of n-hexadecane in soil.The effects of lactose, lactate, vitamins and free amino acids were tested in combinations according to a full factorial design experiment, at concentrations corresponding to those present in fermented whey. The target substance was ¹⁴C-labeled n-hexadecane in nutrient amended soil microcosms contaminated with 5000 mg diesel fuel kg⁻¹ dw. Biodegradation was monitored by determination of evolved ¹⁴CO₂.Significant effects on the biodegradation of n-hexadecane were observed for lactate and amino acids additions in a sandy soil. Lactate showed both an inhibitory effect in the early phase of the experiment and a stimulatory effect in the later phase. The effect of amino acids was slightly stimulatory, mainly evident as a shortening of the lag time.The degree of n-hexadecane degradation at the end of the experiment was correlated with the total concentration of organic compounds added to the soil.

Scientific relevance

There are a handful papers describing the potential of using organic amendments (often industrial by-products) with a content of both easily accessible carbon and micronutrients, to enhance the bioremediation of polluted soils. Enhanced biodegradation is often reported and the proposed explanations are that the combination of easily accessible carbon and micronutrients increases the degrading biomass.In this paper, we examine the effect of fermented whey on the degradation of n-hexadecane and correlate the observed effects on the biodegradation with the main components lactate, amino acids, lactose and B-vitamins. This has to our knowledge never been done before.     

Anaerobic biodegradation of phenolic compounds in digested sludge.

We examined the anaerobic degradation of phenol and the ortho, meta, and para isomers of chlorophenol, methoxyphenol, methylphenol (cresol), and nitrophenol in anaerobic sewage sludge diluted to 10% in a mineral salts medium. Of the 12 monosubstituted phenols studied, only p-chlorophenol and o-cresol were not significantly degraded during an 8-week incubation period. The phenol compounds degraded and the time required for complete substrate disappearance (in weeks) were: phenol (2), o-chlorophenol (3), m-chlorophenol (7), o-methoxyphenol (2), m- and p-methoxyphenol (1), m-cresol (7), p-cresol (3), and o-, m-, and p-nitrophenol (1). Complete mineralization of phenol, o-chlorophenol, m-cresol, p-cresol, o-nitrophenol, p-nitrophenol, and o-, m-, and p-methoxyphenol was observed. In general, the presence of Cl and NO2 groups on phenols inhibited methane production. Elimination or transformation of these substituents was accompanied by increased methane production, o-Chlorophenol was metabolized to phenol, which indicated that dechlorination was the initial degradation step. The methoxyphenols were transformed to the corresponding dihydroxybenzene compounds, which were subsequently mineralized.     

Anaerobic biodegradation of cyanide under methanogenic conditions.

Upflow, anaerobic, fixed-bed, activated charcoal biotreatment columns capable of operating at free cyanide concentrations of greater than 100 mg liter-1 with a hydraulic retention time of less than 48 h were developed. Methanogenesis was maintained under a variety of feed medium conditions which included ethanol, phenol, or methanol as the primary reduced carbon source. Under optimal conditions, greater than 70% of the inflow free cyanide was removed in the first 30% of the column height. Strongly complexed cyanides were resistant to removal. Ammonia was the nitrogen end product of cyanide transformation. In cell material removed from the charcoal columns, [14C]bicarbonate was the major carbon end product of [14C]cyanide transformation.     

Anaerobic biodegradation of pentachlorophenol by a methanogenic consortium

An anaerobic consortium degrading pentachlorophenol (PCP) by methanogenic fermentation was enriched from PCP-contaminated soils. In a semi-continuous reactor, PCP biodegradation was unstable and necessitated periodic additions of unacclimated anaerobic sludge waste to restore the activity. In continuous-flow reactors, PCP degradation activity was more stable when a mixture of glucose and sodium formate was used as secondary carbon source instead of glucose. The analysis of the chlorophenol intermediates suggested that the main pathway of PCP dechlorination was PCP 2,3,5,6-tetrachlorophenol 2,3,5-trichlorophenol 3,5-dichlorophenol 3-chlorophenol phenol. In a laboratory-scale continuous-upflow fixed-film column reactor, a PCP removal of more than 99% was achieved at a PCP loading rate of 60 mol (1 reactor volume)^–1 day^–1 for a hydraulic retention time of 0.7 day. Analysis of culture samples taken at different levels in the reactor have shown that, at this PCP loading rate, only the lower part of the reactor was active. 3-chlorophenol and 3,5- and 3,4-dichlorophenol were detected at the different levels of the reactor. A study of the microorganisms in the biofilm was carried out by scanning electron microscopy and suggested that the microorganisms involved in the consortium were present as a well-structured arrangement. Methanosaeta-like microorganisms were observed mainly at the base of the biofilm whereas, at the surface, a larger diversity of morphotypes was observed in which coccoid or small rod organisms were dominant. This work shows the importance of the design and the control of the operation parameters on the efficiency of the fixed-film reactor.     

Metabolic responses of microbiota to diesel fuel addition in vegetated soil

The effects of trees and contamination on microbial metabolic activity, especially that of hydrocarbon degrading bacteria, were compared during phytoremediation to find which conditions increase diesel fuel removal. Diesel fuel utilisation, microbial extracellular enzyme activities and utilisation of Biolog ECO plate carbon sources by soil bacteria were determined during phytoremediation experiments consisting of two separate diesel applications. Diesel fuel removal after 28 days of second diesel application was 20–30% more than after the first application 1 year earlier. Soil microbiota utilised 26–31 of the 31 Biolog ECO plate carbon sources. Carbon source utilisation profiles indicated minor differences in microbiota in soil vegetated with pine compared to microbiota in soil vegetated with poplar. The potential maximum rates of aminopeptidase activity were 10–10² M AMC/h/g dry soil prior to and after second diesel application, except 14days after the second diesel addition, where the rates were at the scale of 10³M AMC/h/g dry soil. The potential maximum rates of esterase activity were 10³–10⁴M MUF/h/g dry soil. The presence of plants did not influence the activity of esterases. The utilisation of diesel by soil bacteria in Biolog MT2 plate assay was higher in contaminated soil, especially when vegetated, than in uncontaminated soil, measured both as lag times and maximum specific utilisation rates. MT2 plate assay detected the biological response after diesel fuel addition better than general activity methods.     

Anaerobic biodegradation of chlorophenols in fresh and acclimated sludge.

We investigated the anaerobic biodegradation of mono- and dichlorophenol isomers by fresh (unacclimated) sludge and by sludge acclimated to either 2-chlorophenol, 3-chlorophenol, or 4-chlorophenol. Biodegradation was evaluated by monitoring substrate disappearance and, in selected cases, production of 14CH4 from labeled substrates. In unacclimated sludge, each of the monochlorophenol isomers was degraded. The relative rates of disappearance were in this order: ortho greater than meta greater than para. For the dichlorophenols in unacclimated sludge, reductive dechlorination of the Cl group ortho to phenolic OH was observed, and the monochlorophenol compounds released were subsequently degraded. 3,4-Dichlorophenol and 3,5-dichlorophenol were persistent. Sludge acclimated to 2-chlorophenol cross-acclimated to 4-chlorophenol but did not utilize 3-chlorophenol. This sludge also degraded 2,4-dichlorophenol. Sludge acclimated to 3-chlorophenol cross-acclimated to 4-chlorophenol but not to 2-chlorophenol. This sludge degraded 3,4- and 3,5-dichlorophenol but not 2,3- or 2,5-dichlorophenol. The specific cross-acclimation patterns observed for monochlorophenol degradation demonstrated the existence of two unique microbial activities that were in turn different from fresh sludge. The sludge acclimated to 4-chlorophenol could degrade all three monochlorophenol isomers and 2,4- and 3,4-dichlorophenol. The active microbial population in this sludge appeared to be a mixture of populations present in the 2-chlorphenol- and 3-chlorophenol-acclimated sludges, both of which could utilize 4-chlorophenol. Experiments with 14C-radiolabeled p-chlorophenol, o-chlorophenol, and 2,4-dichlorophenol demonstrated that these compounds were converted to 14CH4 and 14CO2.     

A model study of pesticide biodegradation in soil

This study addresses the efficiency of microbial preparations to degrade pesticide residues in soil. A method to degrade pesticides DNOC and pendimethalin using Pseudomonas and Arthrobacter bacteria with a fertilizer is described.     

Estimation of the migration potential of diesel fuel constituents from soil to ground water: A case study

A methodology for estimating the migration potential of diesel fuel constituents from soil to ground water was developed for a large commercial property impacted by diesel fuel. The diesel fuel impacts are associated with former railyard practices that occurred prior to 1970. The site is being redeveloped for commercial use. The proposed improvements for the site include an asphalt‐paved parking lot over the location of the diesel fuel‐impacted soils. The methodology is based on the composition of weathered diesel fuel and the migration characteristics and toxicity data of the diesel fuel constituents. Based on these considerations, the two elements of the methodology are (1) an evaluation of the migration potential of diesel fuel constituents in soil using the soil synthetic rainwater leachate laboratory analysis; and (2) a health‐risk assessment of the diesel fuel ground water impacts. This approach provided sufficient site‐specific data to support a regulatory agency decision allowing development to continue without active remediation of the site soils. If the methodology had not been applied to the site, a remedial method based on a 100 mg/kg to 1000 mg/kg TPH underground storage tank (UST) program soil cleanup level would have likely been required. Considering the project's time constraints and financial requirements, remedial options such as offsite disposal or onsite thermal treatment would have been used resulting in cleanup costs likely exceeding $500, 000. The potential value of this methodology can be best appreciated considering that, based on EPA estimates, there are approximately 295, 000 contaminated UST sites and a significant portion of these sites are contaminated with diesel fuel. These sites could benefit considerably from this approach.     

Anaerobic biodegradation of spent sulphite liquor in a UASB reactor

Anaerobic biodegradation of fermented spent sulphite liquor, SSL, which is produced during the manufacture of sulphite pulp, was investigated. SSL contains a high concentration of lignin products in addition to hemicellulose and has a very high COD load (173 g COD l(-1)). Batch experiments with diluted SSL and pretreated SSL indicated a potential of 12-22 l methane per litre SSL, which corresponds to 0.13-0.22 l methane (g VS)(-1) and COD removal of up to 37%. COD removal in a mesophilic upflow anaerobic sludge blanket, UASB. reactor ranged from 10% to 31% at an organic loading rate, OLR, of 10-51 g (1 d)(-1) and hydraulic retention time from 3.7 to 1.5 days. The biogas productivity was 3 1 (l(reactor d)(-1), with a yield of 0.05 l gas (g VS)(-1). These results suggest that anaerobic digestion in UASB reactors may provide a new alternative for the treatment of SSL to other treatment strategies such as incineration. Although the total COD reduction achieved is limited, bioenergy is produced and readily biodegradable matter is removed causing less load on post-treatment installations.     

Influence of oxygen on the degradation of diesel fuel in soil bioreactors

In fixed-bed bioreactors, the influence of the oxygen content in the inlet air on the biodegradation of diesel fuel in unsaturated soil/compost mixtures was analyzed at 30°C over a period of 7 weeks. Firstly, a wide range from 0 to 80 vol.% O₂ was investigated. Subsequently, the range below 5 vol.% O₂ was examined more closely. Over the whole test period of seven weeks, no significant influence of oxygen could be observed above 1 vol. % O₂ in the inlet air - either on the decrease of the total contaminants or on the total mineralization. Anaerobic conditions should be avoided for the degradation of diesel fuel. During the test period, the courses of CO₂ production varied significantly depending on oxygen supply. Furthermore, a model was developed to estimate the total mineralization as a function of oxygen supply. More investigations are recommended in order to test this model for practical application.     

Anaerobic biodegradation of aromatic hydrocarbons: pathways and prospects

Aromatic hydrocarbons contaminate many environments worldwide, and their removal often relies on microbial bioremediation. Whereas aerobic biodegradation has been well studied for decades, anaerobic hydrocarbon biodegradation is a nascent field undergoing rapid shifts in concept and scope. This review presents known metabolic pathways used by microbes to degrade aromatic hydrocarbons using various terminal electron acceptors; an outline of the few catabolic genes and enzymes currently characterized; and speculation about current and potential applications for anaerobic degradation of aromatic hydrocarbons.     

Anaerobic biodegradation of pentachlorophenol in a contaminated soil inoculated with a methanogenic consortium or with Desulfitobacterium frappieri strain PCP-1

Anaerobic biodegradation of pentachlorophenol (PCP) in a contaminated soil from a wood-treating industrial site was studied in soil slurry microcosms inoculated with a PCP-degrading methanogenic consortium. When the microcosms containing 10%–40% (w/v) soil were inoculated with the consortium, more than 90% of the PCP was removed in less than 30 days at 29 °C. Less-chlorinated phenols, mainly 3-chlorophenol were slowly degraded and accumulated in the cultures. Addition of glucose and sodium formate to the microcosms was not necessary, suggesting that the organic compounds in the soil can sustain the dechlorinating activity. Inoculation of Desulfitobacterium frappieri strain PCP-1 along with a 3-chlorophenol-degrading consortium in the microcosms also resulted in the rapid dechlorination of PCP and the slow degradation of 3-chlorophenol. Competitive polymerase chain reaction experiments showed that PCP-1 was present at the same level throughout the 21-day biotreatment. D. frappieri, strain PCP-1, inoculated into the soil microcosms, was able to remove PCP from soil containing up to 200 mg PCP/kg soil. However, reinoculation of the strain was necessary to achieve more than 95% PCP removal with a concentration of 300 mg and 500 mg PCP/kg soil. These results demonstrate that D. frappieri strain PCP-1 can be used effectively to dechlorinate PCP to 3-chlorophenol in contaminated soils. Received: 14 November 1997 / Received revision: 29 January 1998 / Accepted: 24 February 1998