Showing papers in "Biodegradation in 1999"

Two-liquid-phase bioreactors for enhanced degradation of hydrophobic/toxic compounds.

Abstract: Two-liquid-phase culture systems involve the addition of a water-immiscible, biocompatible and non-biodegradable solvent to enhance a biocatalytic process. Two-liquid-phase bioreactors have been used since the mid-seventies for the microbial and enzymatic bioconversion of hydrophobic/toxic substrates into products of commercial interest. The increasing popularity of bioremediation technologies suggests a new area of application for this type of bioreactor. The toxicity and the limited bioavailability of many pollutants are important obstacles that must first be overcome in order to improve biodegradation processes. Two-liquid-phase bioreactors have the potential to resolve both limitations of biotreatment technologies by the enhancement of the mass-transfer rate of compounds with low bioavailability, and by the controlled delivery of apolar toxic compounds. This technology can also be useful in accelerating the enrichment of microorganisms degrading problematic pollutants. In this paper, we discuss the application of two-liquid-phase bioreactors to enhance the biodegradation of toxic/poorly bioavailable contaminants. Important microbial mechanisms involved in this type of system are described. Uptake of the substrates can be achieved by microorganisms freely dispersed in the aqueous phase and/or bound at the interface between the aqueous and the immiscible phases. Production of surface-active compounds and adhesion abilities are microbial features involved in the process. General guidelines for the design of two-liquid-phase bioreactors for biodegradation purposes are presented. Solvent selection should be established on specific criteria, which depend on the characteristics of target compound(s) and the microorganism(s) implicated in the biodegradation process. The central importance of maximizing the interfacial surface area is highlighted. The potential of this approach as an alternative to current biotreatment technologies is also discussed.

Extracellular oxidative enzyme production and PAH removal in soil by exploratory mycelium of white rot fungi.

Abstract: Selected strains of three species of white rot fungi, Pleurotus ostreatus, Phanerochaete chrysosporium and Trametes versicolor, were grown in sterilized soil from straw inocula The respective colonization rates and mycelium density values decreased in the above mentioned order Three- and four-ringed PAHs at 50 ppm inhibited growth of fungi in soil to some extent The activities of fungal MnP and laccase (units per g dry weight of straw or soil), extracted with 50 mM succinate-lactate buffer (pH 45), were 5 to 20-fold higher in straw compared to soil The enzyme activities per g dry soil in P ostreatus and T versicolor were similar, in contrast to P chrysosporium, where they were extremely low Compared to the aerated controls, P ostreatus strains reduced the levels of anthracene, pyrene and phenanthrene by 81–87%, 84–93% and 41–64% within 2 months, respectively During degradation of anthracene, all P ostreatus strains accumulated anthraquinone PAH removal rates in P chrysosporium and T versicolor soil cultures were much lower

Anaerobic biodegradation of BTEX and gasoline in various aquatic sediments.

Abstract: We examined the extent of biodegradation of benzene, toluene, ethylbenzene and the three isomers of xylene (BTEX) as a mixture and from gasoline in four different sediments: the New York/New Jersey Harbor estuary (polluted); Tuckerton, N.J. (pristine); Onondaga Lake, N.Y. (polluted) and Blue Mtn. Lake, N.Y. (pristine). Enrichment cultures were established with each sediment using denitrifying, sulfidogenic, methanogenic and iron reducing media, as well as site water. BTEX loss, as measured by GC-FID, was extensive in the sediments which had a long history of pollution, with all compounds being utilized within 21–91 days in the most active cultures, and was very slight or non-existent in the pristine sediments. Also, the pattern of loss was different under the various reducing conditions within each sediment and between sediments. For example benzene loss was only observed in sulfidogenic cultures from the NY/NJ Harbor sediments while toluene was degraded under all redox conditions. The loss of BTEX was correlated to the reduction of the various electron acceptors. In cultures amended with gasoline the degradation was much slower and incomplete. These results show that the fate of the different BTEX components in anoxic sediments is dependent on the prevailing redox conditions as well as on the characteristics and pollution history of the sediment.

Efficiency of defined strains and of soil consortia in the biodegradation of polycyclic aromatic hydrocarbon (PAH) mixtures.

Abstract: The microbiological characteristics of the bacterialdegradation of mixtures of five polycyclic aromatichydrocarbons (PAH), phenanthrene, fluorene,anthracene, fluoranthene and pyrene, wereinvestigated. Three pure bacterial strains using oneor several of these PAH as carbon sources wereselected. The interactions between PAH during thedegradation of PAH pairs by each of these strains werestudied and their effects on the kinetics and thebalance of degradation were characterised. Competitionbetween PAH and degradation by cometabolism werefrequently observed. Mixed cultures of two or threestrains, although possessing the global capacity tomineralise the set of five PAH, achieved limiteddegradation of the mixture. In contrast, a consortiumfrom a PAH-contaminated soil readily mineralised thefive-PAH mixture. The results suggested that soilconsortia possessed a wider variety of strains capableto compensate for the competitive inhibition betweenPAH as well as specialised strains that mineralisedpotentially inhibitory PAH metabolites produced bycometabolism.

Degradation of polycyclic aromatic hydrocarbons with three to seven aromatic rings by higher fungi in sterile and unsterile soils.

Abstract: Seven commercial 3- to 7-ring (R) polycyclic aromatic hydrocarbons (PAH) as well as PAH derived from lignite tar were spiked into 3 soils (08 to 97% of organic carbon) The disappearance of the original PAH was determined for the freshly spiked soils, for soils incubated for up to 287 d with their indigenous microflora, and for autoclaved, unsterile and pasteurized soils inoculated with basidiomycetous and ascomycetous fungi Three to 12 d after spiking, 22 to 38% of the PAH could no longer be recovered from the soils At 287 d, 885 to 927%, 834 to 874%, and 220 to 421% of the 3-, 4-, and 5- to 7-R PAH, respectively, had disappeared from the unsterile, uninoculated soils In 2 organic-rich sterile soils, the groups of wood- and straw-degrading, terricolous, and ectomycorrhizal fungi reduced the concentration of 5 PAH by 126, 379, and 94% in 287 d Five- to 7-R PAH were degraded as efficiently as most of the 3- to 4-R PAH In organic-rich unsterile soils inoculated with wood- and straw-degrading fungi, the degradation of 3- to 4-R PAH was not accelerated by the presence of fungi The 5- to 7-R PAH, which were not attacked by bacteria, were degraded by fungi to 29 to 42% in optimum combinations of fungal species and soil type In organic-poor unsterile soil, these same fungi delayed the net degradation of PAH possibly for 2 reasons Mycelia of Pleurotus killed most of the indigenous soil bacteria expected to take part in the degradation of PAH, whereas those of Hypholoma and Stropharia promoted the development of opportunistic bacteria in the soil, which must not necessarily be PAH degraders Contemporarily, the contribution of the fungi themselves to PAH degradation may be negligible in the absence of soil organic matter due to the lower production of ligninolytic enzymes It is concluded that fungi degrade PAH irrespective of their molecular size in organic-rich and wood chip-amended soils which promote fungal oxidative enzyme production

Analyses of Polycyclic Aromatic Hydrocarbon Degrading Bacteria Isolated from Contaminated Soils

Abstract: Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria isolated from PAH-contaminated soils were analyzed genotypically and phenotypically for their capacity for metabolism of naphthalene and other PAH substrates. The methods used for the analyses were DNA hybridization using NAH7-derived gene probes, PAH spray plate assays, 14C-PAH mineralization assays, and dioxygenase activity assays. The results of the analyses showed a dominant number of PAH-degrading bacteria with a NAH7-like genotype. The results support the continued use of the nahA probe for contaminated soils to monitor the genetic potential of indigenous microorganisms to degrade PAHs. However, the finding of non-it nahA-hybridizing PAH-degrading bacteria show the limitation of NAH7-derived gene probes. Fifteen percent (13/89) of PAH-degrading bacteria isolated were not detected with the nahA gene probe. Four isolates (designated A5PH1, A8AN3, B1PH2, and B10AN1) did not hybridize with any of the NAH7-derived gene probes ( nahA, nahG, nahH, and nahR) used in this study. Considering the numerous unculturable microorganisms in nature and their potential genotypes, NAH7-derived gene probes may underestimate the microbial potential to catabolize PAHs. This necessitates development of new gene probes for enumeration and isolation of PAH-degrading bacteria to better understand the in situ microbial potential to degrade PAHs.

Degradation of mono-chlorophenols by a mixed microbial community via a meta- cleavage pathway.

Abstract: A mixed microbial community, specially designed todegrade a wide range of substituted aromaticcompounds, was examined for its ability to degrademono-chlorophenols as sole carbon source in aerobicbatch cultures. The mixed culture degraded 2-, 3-, and4 -chlorophenol (1.56 mM) via a meta- cleavagepathway. During the degradation of 2- and3-chlorophenol by the mixed culture, 3-chlorocatecholproduction was observed. Further metabolism was toxicto cells as it led to inactivation of the catechol2,3-dioxygenase enzyme upon meta- cleavage of3-chlorocatechol resulting in incomplete degradation.Inactivation of the meta- cleavage enzyme led toan accumulation of brown coloured polymers, whichinterfered with the measurement of cell growth usingoptical denstiy. Degradation of 4-chlorophenol by themixed culture led to an accumulation of5-chloro-2-hydroxymuconic semialdehyde, themeta- cleavage product of 4-chlorocatechol. Theaccumulation of this compound did not interfere withthe measurement of cell growth using optical density.5-chloro-2-hydroxymuconic semialdehyde was furthermetabolized by the mixed culture with a stoichiometricrelease of chloride, indicating complete degradationof 4-chlorophenol by the mixed culture via ameta- cleavage pathway.

Transformation and mineralization of 2,4,6-trinitrotoluene (TNT) by manganese peroxidase from the white-rot basidiomycete Phlebia radiata.

Abstract: The degradation of the nitroaromatic pollutant 2,4,6-trinitrotoluene (TNT) by the manganese-dependent peroxidase (MnP) of the white-rot fungus Phlebia radiata and the main reduction products formed were investigated. In the presence of small amounts of reduced glutathione (10 mM), a concentrated cell-free preparation of MnP from P. radiata exhibiting an activity of 36 nkat/ml (36 nmol Mn(II) oxidized per sec and per ml) transformed 10 mg/l of TNT within three days. The same preparation was capable of completely transforming the reduced derivatives of TNT. When present at 10 mg/l, the aminodinitrotoluenes were transformed in less than two days and the diaminonitrotoluenes in less than three hours. Experiments with 14C-U-ring labeled TNT and 2-amino-4,6-dinitrotoluene showed that these compounds were mineralized by 22% and 76%, respectively, within 5 days. Higher concentrations of reduced glutathione (50 mM) led to a severe inhibition of the degradation process. It is concluded that Phlebia radiata is a good candidate for the biodegradation of TNT as well as its reduction metabolites.

A new test procedure for biogenic sulfuric acid corrosion of concrete

Abstract: A new test method is described for biogenic sulfuric acid corrosion of concrete, more specifically in sewer conditions. The aim of the new test method is the development of an accelerated and reproducible procedure for monitoring the resistance of different types of concrete with regard to biogenic sulfuric acid corrosion. This experimental procedure reflects worst case conditions by providing besides H2S, also an enrichment of thiobacilli and biologically produced sulfur. By simulating the cyclic processes occurring in sewer pipes, significant differences between concrete mixtures could be detected after 51 days. Concrete modified by a styrene-acrylic ester polymer demonstrated a higher resistance against biogenic sulfuric acid attack.

Intrinsic bioremediation of a petroleum hydrocarbon-contaminated aquifer and assessment of mineralization based on stable carbon isotopes.

Abstract: This study presents a stepwise concept to assess the in situ microbial mineralization of petroleum hydrocarbons (PHC) in aquifers. A new graphical method based on stable carbon isotope ratios (δ13C) was developed to verify the origin of dissolved inorganic carbon (DIC). The concept and the isotope method were applied to an aquifer in Studen, Switzerland, in which more than 34,000 liters of heating oil were accidentally released. Chemical analyses of ground water revealed that in this aquifer locally, anaerobic conditions prevailed, and that PHC mineralization was linked to the consumption of oxidants such as O2, NO2-, and SO42- and the production of reduced species such as Fe2+, Mn2+, HSS and CH4. However, alkalinity and DIC balances showed a quantitative disagreement in the link between oxidant consumption and DIC production, indicating that chemical data alone may not be a reliable assessment tool. δ13C ratios in DIC have been used before for bioremediation assessment, but results were reported to be negatively influenced by methanogenesis. Using the new graphical method to display δ13C data, it was possible to identify anomalies found in methanogenic monitoring wells. It could be shown that 88% of the DIC produced in the contaminated aquifer originated from microbial PHC mineralization. Thus, the new graphical method to display δ13C ratios appears to be a useful tool for the assessment of microbial hydrocarbon mineralization in a complex environment.

Degradation of anaerobic reductive dechlorination products of Aroclor 1242 by four aerobic bacteria

Abstract: We studied the aerobic degradation of eight PCB congeners which comprise from 70 to 85% of the anaerobic dechlorination products from Aroclor 1242, including 2-, 4-, 2,4-, 2,6-, 2,2'-, 2,4'-, 2,2', 4-, and 2,4,4'-chlorobiphenyl (CB), and the biodegradation of their mixtures designed to simulate anaerobic dechlorination profiles M and C. Strains Comamonas testosteroni VP44 and Rhodococcus erythreus NY05 preferentially oxidized a para-substituted ring, while Rhodococcus sp. RHA1, similar to well known strain Burkholderia sp. LB400, preferably attacked an ortho-chlorinated ring. Strains with ortho-directed attack extensively degraded 2,4'- and 2,4,4'-CB into 4-chlorobenzoate, while bacteria with para-directed attack transformed these congeners mostly into potentially problematic meta-cleavage products. The strains that preferentially oxidized an ortho-substituted ring readily degraded seven of the eight congeners supplied individually; only 2,6-CB was poorly degraded. Degradation of 2,2'- and 2,4,4'-CB was reduced when present in mixtures M and C. Higher efficiencies of degradation of the individual congeners and defined PCB mixtures M and C and greater production of chlorobenzoates were observed with bacteria that preferentially attack an ortho-substituted ring. PCB congeners 2,4'-, 2,2',4-, and 2,4,4'-CB can be used to easily identify bacteria with ortho-directed attack which are advantageous for use in the aerobic stage of the two-phase (anaerobic/aerobic) PCB bioremediation scheme.

Anaerobic biodegradability of phthalic acid isomers and related compounds.

Abstract: All three phthalic acid isomers ( ortho, meta and para benzene dicarboxylic acid) are produced in massive amounts, and used in the chemical industry as plasticizers or for the production of polyester. Wastestreams generated during the production of phthalate isomers generally contain high concentrations of aromatic acids. To study the potential biodegradability of these primarily anthropogenic compounds in anaerobic bioreactors, biodegradability studies were performed. Compounds tested were benzoate, ortho-phthalate, isophthalate, terephthalate, dimethyl phthalate, dimethyl terephthalate, para-toluate and para-xylene. Seed materials tested were two types of granular sludge and digested sewage sludge. It was found that all phthalate isomers and their corresponding dimethyl-esters, could be completely mineralized by all seed materials studied. Lag phases required for 50% degradation of these compounds, ranged from 17 to 156 days. The observed degradation curves could be explained by growth of an initially small amount of organisms in the inoculum with the specific ability to degrade one phthalate isomer. The observed order in the length of the lag phases for the phthalate isomers is: phthalate < terephthalate < isophthalate. This order appears to be related to the environmental abundancy of the different phthalate isomers. The initial step in the degradation pathway of both dimethyl phthalate esters was hydrolysis of the ester sidechain, resulting in the formation of the corresponding mono-methyl-phthalate isomer and phthalate isomer. The rate limiting step in mineralization of both dimethyl phthalate and dimethyl terephthalate was found to be fermentation of the phthalate isomer. Para-toluate was degraded only by digested sewage sludge after a lag phase of 425 days. The observed degradation rates of this compound were very low. No mineralization of para-xylene was observed. In general, the differences in the lag phases between different seed materials were relatively small. These results indicate that the time needed for the start-up of anaerobic bioreactors treating wastewaters containing phthalic acid isomers, depends little on the microbial composition of the seed material applied, but may take several months.

Facilitation of cometabolic degradation of 4-chlorophenol using glucose as an added growth substrate.

Abstract: This paper reports on the feasibility of using glucose as an added substrate for cometabolic transformation of 4-chlorophenol (4-cp). When glucose was fed as the added growth substrate, only 78% and 43% of the initial 4-cp concentrations of 100 and 200 mg l-1, respectively, were transformed before the pH dropped to below 4.5 and stopped all reactions. By maintaining the medium pH, complete removal of 4-cp was achieved even at the high initial concentration of 200 mg l-1. Phenol induction prior to inoculation was not a prerequisite to ensure transformation of 4-cp when glucose was the added growth substrate. Compared with phenol as the added growth substrate, cells grown on glucose displayed a longer acclimation phase and, in general, a lower specific transformation rate. The volumetric transformation rate of 4-cp, however, was greatly enhanced due to the increased cell density. The results of this work suggest that 4-cp itself induced the enzymes necessary for its cometabolism. With NADH regenerated effectively through metabolism of glucose, 4-cp was transformed in the absence of added phenol. Consequently, the competitive inhibition involved in cometabolism was avoided and the risks associated with addition of toxic growth substrates such as phenol were eliminated.

Biodegradation of 2-methyl, 2-ethyl, and 2-hydroxypyridine by an Arthrobacter sp. isolated from subsurface sediment.

Abstract: A bacterium capable of degrading 2-methylpyridine was isolated by enrichment techniques from subsurface sediments collected from an aquifer located at an industrial site that had been contaminated with pyridine and pyridine derivatives. The isolate, identified as an Arthrobacter sp., was capable of utilizing 2-methylpyridine, 2-ethylpyridine, and 2-hydroxypyridine as primary C, N, and energy sources. The isolate was also able to utilize 2-, 3-, and 4-hydroxybenzoate, gentisic acid, protocatechuic acid and catechol, suggesting that it possesses a number of enzymatic pathways for the degradation of aromatic compounds. Degradation of 2-methylpyridine, 2-ethylpyridine, and 2-hydroxypyridine was accompanied by growth of the isolate and release of ammonium into the medium. Degradation of 2-methylpyridine was accompanied by overproduction of riboflavin. A soluble blue pigment was produced by the isolate during the degradation of 2-hydroxypyridine, and may be related to the diazadiphenoquinones reportedly produced by other Arthrobacter spp. when grown on 2-hydroxypyridine. When provided with 2-methylpyridine, 2-ethylpyridine, and 2-hydroxypyridine simultaneously, 2-hydroxypyridine was rapidly and preferentially degraded; however there was no apparent biodegradation of either 2-methylpyridine or 2-ethylpyridine until after a seven day lag. The data suggest that there are differences between the pathway for 2-hydroxypyridine degradation and the pathway(s) for 2-methylpyridine and 2-ethylpyridine.

Population dynamics of an introduced bacterium degrading chlorinated benzenes in a soil column and in sewage sludge.

Abstract: The capacity of the β-Proteobacterium Pseudomonas sp. strain P51, which degrades chlorinated benzenes, to metabolize 1,2,4-trichlorobenzene (TCB) under environmental conditions was tested by its release into two experimental systems. The first system consisted of laboratory scale microcosms which were operated with and without the addition of TCB and which were inoculated with sludge from a wastewater treatment plant. The second system consisted of a non sterile, water saturated soil column. We determined survival of strain P51 after its introduction and its ability to degrade TCB. The population dynamics was followed by selective plating and applying the polymerase chain reaction (PCR) to detect strain P51 and the chlorobenzene ( tcb) genes on catabolic plasmid pP51. The results showed a completely different behaviour of strain P51 in the two habitats under the applied conditions. In the soil column the P51 bacteria inoculated the entire area and their population reached 2 × 106 cells/g soil. The population remained active since TCB was degraded to concentrations below the detection limit of 30 μg/l. In the sludge microcosms, the number of strain P51 cells immediately decreased from 4 × 107 cells/ml to 105 cells/ml over a period of 2 days after inoculation, and then the strain disappeared to levels below our detection limit (103–104 cells/ml). In the reactor without TCB the population of P51 maintained a stable value of 105 cells/ml during 8 days but then also decreased to levels below the detection limit. In addition, no significant TCB degradation was found in the sludge reactors. The influence of presence of TCB on maintenance of strain P51 in the two habitats is discussed. This work demonstrates the possibility to successfully apply preselected strains to degrade otherwise poorly degradable substances in complex mixed microbial communities. However, survival and activity may depend strongly on the type of system into which the strain is introduced.

Biodegradation of gasoline and BTEX in a microaerophilic biobarrier.

Abstract: Continuous bioremediation of gasoline-contaminatedwater in a packed-bed biobarrier system underoxygen-limited conditions is discussed. This studywas part of an extensive effort to develop analternative technology for the in situbioremediation of hydrocarbons where there is alimited supply of oxygen. Protruded stainless steelpieces and granulated peat moss were used as packingmaterial to support microbial growth in twobiobarriers. The inoculum was an enrichment culture ofan indigenous microbial population from a soil sample.The biobarriers' inlet gasoline concentrations and thelinear liquid velocities were similar to thosecommonly found at in situ conditions. Gasolineremoval efficiencies ranged from 94% to 99.9% in thestainless steel-packed biobarrier, and from 86.6% to99.6% in the peat moss-packed biobarrier. Effluentgasoline concentrations below 0.03 mg/l were obtainedat gasoline loading rates less than 27.5 mg/l.d in thestainless steel-packed biobarrier. The remainingfraction of gasoline in the effluent consisted mainlyof three aliphatic compounds and not the aromaticcompounds. Both biobarrier packings supported nearcomplete removal of the most soluble aromatichydrocarbons of gasoline (BTEX) under all theconditions examined. The consumption of sulfate andthe presence of sulfate-reducing microorganismssuggested the presence of anaerobic metabolism duringthe degradation of gasoline. Up to 92% gasoline wasremoved during the first 3 cm of the biobarriers'length.

Effect of nitrogen and phosphorus addition on phenanthrene biodegradation in four soils.

Abstract: Phenanthrene mineralization rates were found to vary widely among four soils; differences in soil nutrient levels was one hypothesis to explain this variation To test this hypothesis, phenanthrene mineralization rates were measured in these soils with, and without, added nitrogen and phosphorus Mineralization rates either remained unchanged or were depressed by the addition of nitrogen and phosphorus Phenanthrene degradation rates remained unchanged in the soil which had the highest indigenous levels of nitrogen and phosphorus and which showed the largest increase in phosphorus levels after nutrients were added The soils in which degradation rates were depressed had lower initial phosphorus concentrations and showed much smaller or no measurable increase in phosphorus levels after nutrients were added to the soils To understand the response of phenanthrene degradation rates to added nitrogen and phosphorus, it may be necessary to consider the bioavailability of added nutrients and nutrient induced changes in microbial metabolism and ecology

Fate of the herbicides mecoprop, dichlorprop, and 2,4-D in aerobic and anaerobic sewage sludge as determined by laboratory batch studies and enantiomer-specific analysis.

Abstract: Aerobic degradation experiments with the racemic mixtures of mecoprop and dichlorprop revealed that activated sludge collected from the aeration tank of a municipal waste water treatment plant degraded both enantiomers of mecoprop and dichlorprop within 7 days, albeit in an enantioselective manner; the (S) enantiomers were preferentially degraded. Mecoprop, dichlorprop, and 2,4-D were completely metabolized under aerobic conditions, as shown by the 86–98% elimination of dissolved organic carbon. Under anaerobic conditions, the concentration of 2,4-D decreased exponentially with a first-order reaction rate constant of 0.24 per day and without a lag-phase. After an incubation time of 17 days, 2,4-D was completely removed. 2,4-Dichlorophenol was the main metabolite of anaerobic 2,4-D degradation; only traces of 4-chlorophenol were detected. In contrast, the chiral phenoxypropionic acid herbicides mecoprop and dichlorprop persisted under anaerobic conditions during 49 days of incubation.

Diversity and correlation of specific aromatic hydrocarbon biodegradation capabilities.

Abstract: This work investigated the biodegradation capabilitiesof indigenous microorganisms exposed to differentcombinations of aromatic hydrocarbons. Considerablediversity was found in the catabolic specificity of 55strains. Toluene was the most commonly degradedcompound, followed by p-xylene, m-xyleneand ethylbenzene. Strains capable of degradingo-xylene and benzene, which were theleast-frequently-degraded compounds, exhibited broaderbiodegradation capabilities. Kappa statistics showeda significant correlation between the abilities todegrade toluene and ethylbenzene, p-xylene andm-xylene, and p-xylene and o-xylene. The ability to degrade naphthalene was correlated tothe ability to degrade other alkylbenzenes, but notbenzene. In addition, the inability to degradebenzene was correlated to the inability to degradeo-xylene. Factorial analysis of variance showedthat biodegradation capabilities were generallybroader when aromatic hydrocarbons were fed asmixtures than when fed separately. Beneficialsubstrate interactions included enhanced degradationof benzene, p-xylene, and naphthalene whentoluene was present, and enhanced degradation ofnaphthalene by ethylbenzene. Such heuristicrelationships may be useful to predict biodegradationpatterns when bacteria are exposed to differentaromatic hydrocarbon mixtures.

Modelling the growth of a methanotrophic biofilm: Estimation of parameters and variability

Abstract: This article discusses the growth of methanotrophic biofilms. Several independent biofilm growths scenarios involving different inocula were examined. Biofilm growth, substrate removal and product formation were monitored throughout the experiments. Based on the oxygen consumption it was concluded that heterotrophs and nitrifiers co-existed with methanotrophs in the biofilm. Heterotrophic biomass grew on soluble polymers formed by the hydrolysis of dead biomass entrapped in the biofilm. Nitrifier populations developed because of the presence of ammonia in the mineral medium. Based on these experimental results, the computer program AQUASIM was used to develop a biological model involving methanotrophs, heterotrophs and nitrifiers. The modelling of six independent growth experiments showed that stoichiometric and kinetic parameters were within the same order of magnitude. Parameter estimation yielded an average maximum growth rate for methanotrophs, μm, of 1.5 ± 0.5 d−1, at 20 °C, a decay rate, bm, of 0.24 ± 0.1 d−1, a half saturation constant, \({\text{K}}_{{\text{S(CH}}_{\text{4}} {\text{)}}} \), of 0.06 ± 0.05 mg CH4/L, and a yield coefficient, \(Y_{CH_4 } \), of 0.57 ±: 0.04 g X/g CH4. In addition, a sensitivity analysis was performed on this model. It indicated that the most influential parameters were those related to the biofilm (i.e. density; solid-volume fraction; thickness). This suggests that in order to improve the model, further research regarding the biofilm structure and composition is needed.

Biofilter performance and characterization of a biocatalyst degrading alkylbenzene gases.

Abstract: A biofilter treating alkylbenzene vapors was characterized for its optimal running conditions and kinetic parame-ters. Kinetics of the continuous biofilter were compared to batch kinetic data obtained with biofilm samples as well as with defined microbial consortia and with pure culture isolates from the biofilter. Both bacteria and fungi were present in the bioreactor. Five strains were isolated. Two bacteria, Bacillus and Pseudomonas, were shown to be dominant, as well as a Trichosporon strain which could, however, hardly grow on alkylbenzenes in pure culture. The remaining two strains were most often overgrown by the other three organisms in liquid phase batch cultures μ max, KS, KI values and biodegradation rates were calculated and compared for the difterent mixed and pure cultures. Since filter bed acidification was observed during biofiltration studies reaching a pH of about 4, experiments were also undertaken to study the influence of pH on performance of the different cultures. Biodegradation and growth were possible in all cases, over the pH range 3.5–7.0 at appreciable rates, both with mixed cultures and with pure bacterial cultures. Under certain conditions, microbial activity was even observed in the presence of alkylbenzenes down to pH 2.5 with mixed cultures, which is quite unusual and explains the ability of the present biocatalyst to remove alkylbenzenes with high efficiency in biofilters under acidic conditions.

Photochemical mineralization of synthetic lignin in lake water indicates enhanced turnover of aromatic organic matter under solar radiation

Abstract: The degradation of 14 C-[ring]-labelled synthetic lignin ( 14 C-DHP) and dissolved organic carbon (DOC) from lake water were studied simultaneously. 14 C-DHP was incubated in humic lake water (colour 173 mg Pt l 1 ) for 7 d in the dark or under solar radiation. In the dark <0.4% of the introduced 14 C-DHP label and 4% of the indigenous DOC were mineralized, indicating that the 14 C-labelled aromatic rings of DHP and the humic DOC were microbiologically recalcitrant. Under solar radiation (116 MJ m 2 ), 17‐21% of the 14 C-labelled carbons in DHP and 18‐23% of the indigenous DOC were mineralized in 7 d. Simultaneously the water solubility of 14 C-DHP increased. Solar radiation converted the aromatic cores of synthetic lignin to CO2 and soluble organic photoproducts. The results suggest that solar radiation plays a key role in the decomposition of natural polyaromatic matter.

Modeling speciation effects on biodegradation in mixed metal/chelate systems

Abstract: A new model, CCBATCH, comprehensively couples microbially catalyzed reactions to aqueous geochemistry The effect of aqueous speciation on biodegradation reactions and the effect of biological reactions on the concentration of chemical species (eg H2CO3, NH 4 + , O2) are explicitly included in CCBATCH, allowing systematic investigation of kinetically controlled biological reactions Bulk-phase chemical speciation reactions including acid/base and complexation are modeled as thermodynamically controlled, while biological reactions are modeled as kinetically controlled A dual-Monod kinetic formulation for biological degradation reactions is coupled with stoichiometry for the degradation reaction to predict the rate of change of all biological and chemical species affected by the biological reactions The capability of CCBATCH to capture pH and speciation effects on biological reactions is demonstrated by a series of modeling examples for the citrate/Fe(III) system pH controls the concentration of potentially biologically available forms of citrate When the percentage of the degradable substrate is low due to complexation or acid/base speciation, degradation rates may be slow despite high concentrations of substrate Complexation reactions that sequester substratein non-degradable forms may prevent degradation or stopdegradation reactions prior to complete substrate utilization The capability of CCBATCH to couple aqueous speciation changes to biodegradation reaction kinetics and stoichiometry allows prediction of these key behaviors in mixed metal/chelate systems

Identification of the key factors affecting composting of a weathered hydrocarbon-contaminated soil

Abstract: The effects of the C/N ratio, CaCO3 and PO4 addition, and temperature profile on reactor-based composting of weathered hydrocarbon-contaminated soil were evaluated in a series of 30-day tests in temperature-controlled mini-composters. Soil containing 17,000 mg (kg dry soil)−1 mineral oil and grease (MOG) was composted with maple leaves and alfalfa. Although the leaves and alfalfa also contained MOG, degradation of contaminated soil derived MOG (total MOG degradation minus MOG degradation in a control with no soil) increased from 0 to 45% as the quantity of co-substrate increased from 0 to 63%. Simulation of biopile conditions (i.e., aeration and addition of mineral salts but no co-substrate) resulted in only 6% MOG degradation. Addition of CaCO3 before composting increased total MOG degradation from 23% to 43%. Total MOG degradation increased with decreasing C/N ratio. At a molar C/N ratio of 17, 43% of the total MOG was degraded in 30 days, while at a C/N ratio of 40 there was no total MOG degradation. When temperatures ranging from 23 to 60 °C were investigated, 50 °C maintained for 29 days resulted in the maximum degradation which was 68% of total initial MOG.

Transformation of diphenyl ethers by Trametes versicolor and characterization of ring cleavage products

Abstract: The white-rot fungi Trametes versicolor SBUG 1050, DSM 11269 and DSM 11309 are able to oxidize diphenyl ether and its halogenated derivatives 4-bromo- and 4-chlorodiphenyl ether The products formed from diphenyl ether were 2- and 4-hydroxydiphenyl ether Both 4-bromo- and 4-chlorodiphenyl ether were transformed to the corresponding products hydroxylated at the non-halogenated ring Additionally, ring-cleavage products were detected by high perfomance liquid chromatography and characterized by gas chromatography/mass spectrometry and proton nuclear magnetic resonance spectroscopy Unhalogenated diphenyl ether was degraded to 2-hydroxy-4-phenoxymuconic acid and 6-carboxy-4-phenoxy-2-pyrone Brominated derivatives of both these compounds were formed from 4-bromodiphenyl ether, and 4-chlorodiphenyl ether was transformed in the same way to the analogous chlorinated ring cleavage products Additionally, 4-bromo- and 4-chlorophenol were detected as intermediates from 4-bromo- and 4-chlorodiphenyl ether, respectively In the presence of the cytochrome-P450 inhibitor 1-aminobenzotriazole, no metabolites were formed by cells of Trametes versicolor from the diphenyl ethers investigated Cell-free supernatants of whole cultures with high laccase and manganese peroxidase activities were not able to transform the unhydroxylated diphenyl ethers used

Removal of mercury from mercury-contaminated sediments using a combined method of chemical leaching and volatilization of mercury by bacteria.

Abstract: A method for the removal of mercury sulfide from mercury-contaminated sediments was developed, which consists of chemical leaching and volatilization of mercury by bacteria. More than 85% of the mercury in sediment containing 0.11-37.4 mg/kg of mercury was efficiently extracted with 3 M HCl and 74 mM FeCl3. Subsequent volatilization by bacteria resulted in the removal of 62.9-75.1% of mercury from mercury-contaminated Minamata Bay sediments. Methylmercury was also eliminated from soil at a high efficiency. Thus, this combined method of chemical and microbial treatments could be used for efficient removal of both organic and inorganic mercurials from natural sediments.

Biodegradation of aromatic compounds and TCE by a filamentous bacteria-dominated consortium.

Abstract: The Michaelis-Menten biodegradation kinetics (k and Ks) of aromatic compounds and trichloroethene (TCE) by an aerobic enrichment culture grown on phenol and dominated by a unique filamentous bacterium were measured. The average k and Ks values for phenol, benzene (B), toluene (T), ethylbenzene (E), o-xylene (oX), p-xylene (pX), naphthalene and TCE in g per g VSS-d and mg L-1 were 5.72 and 0.34, 1.20 and 0.51, 2.09 and 0.47, 0.77 and 0.23, 0.61 and 0.16, 0.73 and 0.23, 0.17 and 0.18, and 0.16 and 0.18, respectively. Significant variability in these measured kinetics was noted between tests conducted over the 5-month period during which the fed-batch culture with a 5-day solids retention time was maintained; the coefficient of variation of the k and Ks values ranged from 11–43% and 4–50%, respectively. This variation was significantly greater than the method measurement error on a given date. Degradation of BTEoXpX mixtures could be described by a basic competitive inhibition model.

Biodegradation of xylene and butyl acetate using an aqueous-silicon oil two-phase system.

Abstract: A stable microbial population, consisting of seven bacterial strains and three yeast strains, was selected in batch cultures on a mixture of ortho and meta-xylene and butyl acetate as the sole source of carbon and energy. This population can completely degrade up to 10 g/L of a mixture of these xenobiotics (70% xylene and 30% butyl acetate wt/wt) in a two-phase aqueous-silicone oil system (70%/30% vol/vol) within 96 h, while for the usual one-phase system very low growth degradation rates were observed. Further organic solvents were tested and finally, silicon oil was selected as the best organic phase for such a two-phase system. With periodical pH adjustments to 6.0 in fed-batch mode, the culture showed a global degradation rate of 63 mg L-1 h-1.

Dolomite limits acidification of a biofilter degrading dimethyl sulphide

Abstract: The applicability of dolomite particlesto control acidificationin a Hyphomicrobium MS3inoculated biofilter removingdimethyl sulphide (Me2S) wasstudied. While direct inoculationof the dolomite particles with theliquid microbial culture was notsuccessful, start-up ofMe2S-degradation in thebiofilter was observed when thedolomite particles were mixed with33% (wt/wt) of Hyphomicrobium MS3-inoculatedcompost or wood bark material.Under optimal conditions, anelimination capacity (EC) of 1680~g Me2S m-3 d-1 wasobtained for the compost/dolomitebiofilter. Contrary to a wood barkor compost biofilter, no reductionin activity due to acidificationwas observed in these biofiltersover a 235 day period because ofthe micro environmentneutralisation of the microbialmetabolite H2SO4 with thecarbonate in the dolomite material.However, performance of thebiofilter decreased when themoisture content of the mixedcompost/dolomite material droppedbelow 15%. Next to this, nutrientlimitation resulted in a gradualdecrease of the EC andsupplementation of a nitrogensource was a prerequisite to obtaina long-term high EC (> 250 gMe2S m-3 d-1) forMe2S. In relation to thisnitrogen supplementation, it wasobserved that stable ECs forMe2S were obtained when thisnutrient was dosed to the biofilterat a Me2S-C/NH4Cl-Nratio of about 10.Abbreviations:DW – dry weight,EC – elimination capacity,Me2S – dimethyl sulphide,OL – organic loading rate,VS - volatile solids

Growth inhibition of biomass adapted to the degradation of toluene and xylenes in mixture in a batch reactor with substrates supplied by pulses

Abstract: A biomass adapted to degrade toluene and xylenes in mixture was grown in a batch reactor with substrates supplied by pulses. The inhibition of biomass growth in the course of substrate degradation was investigated. The maximal biomass concentration of 7 g l−1 was obtained using 150 μl of toluene and 15 μl of a mixture of xylenes in one litre of liquid medium, and the maximal biomass productivity and yield were 53 mg l−1 h−1 and 0.32 gDW g s −1 , respectively. Higher quantities of substrate added by pulses, that is 200 μl of toluene with 20 μl of xylenes and 300 μl of toluene with 30 μl of xylenes, caused an accumulation of metabolites. These higher quantities of substrates caused inhibition of microbial growth. Among the metabolites produced, 4-methyl catechol was found in large quantities in the culture medium and in the cells.