Biodegradation of DDT by stimulation of indigenous microbial populations in soil with cosubstrates

Stimulation of native microbial populations in soil by the addition of small amounts of secondary carbon sources (cosubstrates) and its effect on the degradation and theoretical mineralization of DDT [l,l,l-trichloro-2,2-bis(p-chlorophenyl)ethane] and its main metabolites, DDD and DDE, were evaluated. Microbial activity in soil polluted with DDT, DDE and DDD was increased by the presence of phenol, hexane and toluene as cosubstrates. The consumption of DDT was increased from 23 % in a control (without cosubstrate) to 67, 59 and 56 % in the presence of phenol, hexane and toluene, respectively. DDE was completely removed in all cases, and DDD removal was enhanced from 67 % in the control to ~86 % with all substrates tested, except for acetic acid and glucose substrates. In the latter cases, DDD removal was either inhibited or unchanged from the control. The optimal amount of added cosubstrate was observed to be between 0.64 and 2.6 mg C $ {\text{g}}^{ - 1}_{\text{dry soil}} $ . The CO₂ produced was higher than the theoretical amount for complete cosubstrate mineralization indicating possible mineralization of DDT and its metabolites. Bacterial communities were evaluated by denaturing gradient gel electrophoresis, which indicated that native soil and the untreated control presented a low bacterial diversity. The detected bacteria were related to soil microorganisms and microorganisms with known biodegradative potential. In the presence of toluene a bacterium related to Azoarcus, a genus that includes species capable of growing at the expense of aromatic compounds such as toluene and halobenzoates under denitrifying conditions, was detected.     

Biodegradation of 4-nitrophenol by indigenous microbial populations in Everglades soils

The Everglades in South Florida are a unique ecologicalsystem. As a result of the widespread use of pesticides andherbicides in agricultural areas upstream from these wetlands,there is a serious potential for pollution problems in theEverglades. The purpose of this study was to evaluate theability of indigenous microbial populations to degradexenobiotic organic compounds introduced by agricultural andother activities. Such biodegradation may facilitate theremediation of contaminated soils and water in the Everglades.The model compound selected in this study is 4-nitrophenol, achemical commonly used in the manufacture of pesticides. Themineralization of 4-nitrophenol at various concentrations wasstudied in soils collected from the Everglades. Atconcentrations of 10 and 100 µg/g soil, considerablemineralization occurred within a week. At a higherconcentration, i.e., 10 mg/g soil, however, no mineralizationof 4-nitrophenol occurred over a 4-month period; such a highconcentration apparently produced an inhibitory effect. Therate and extent of 4-nitrophenol mineralization was enhancedon inoculation with previously isolated nitrophenol-degradingmicroorganisms. The maximum mineralization extent measured,however, was less than 30% suggesting conversion to biomassand/or unidentified intermediate products. These resultsindicate the potential for natural mechanisms to mitigate theadverse effects of xenobiotic pollutants in a complex systemsuch as the Everglades.     

Biodegradation of 4-nitrophenol by indigenous microbial populations in Everglades soils

The Everglades in South Florida are a unique ecological system. As a result of the widespread use of pesticides and herbicides in agricultural areas upstream from these wetlands, there is a serious potential for pollution problems in the Everglades. The purpose of this study was to evaluate the ability of indigenous microbial populations to degrade xenobiotic organic compounds introduced by agricultural and other activities. Such biodegradation may facilitate the remediation of contaminated soils and water in the Everglades. The model compound selected in this study is 4-nitrophenol, a chemical commonly used in the manufacture of pesticides. The mineralization of 4-nitrophenol at various concentrations was studied in soils collected from the Everglades. At concentrations of 10 and 100 microg/g soil, considerable mineralization occurred within a week. At a higher concentration, i.e., 10 mg/g soil, however, no mineralization of 4-nitrophenol occurred over a 4-month period; such a high concentration apparently produced an inhibitory effect. The rate and extent of 4-nitrophenol mineralization was enhanced on inoculation with previously isolated nitrophenol-degrading microorganisms. The maximum mineralization extent measured, however, was less than 30% suggesting conversion to biomass and/or unidentified intermediate products. These results indicate the potential for natural mechanisms to mitigate the adverse effects of xenobiotic pollutants in a complex system such as the Everglades.     

Biodegradation of BTE in soil by indigenous microbial populations with and without biogenic substrates

Degradation of benzene, toluene, and ethylbenzene (BTE) by microbial populations indigenous to the soil and populations proliferated from the indigenous using biogenic substrates were compared. The reaction system consisted of aerobic microcosms representing an unsaturated soil. Microcosms supplemented with glucose and citrate, when compared to the unsupplemented microcosms, showed increases in bacterial counts, but the overall degradation rates for B, T, or E were reduced in spite of shorter lag times. Both biogenic substrate supplements were non-beneficial for BTE degradation due largely to the preferential and healthy growth of the indigenous populations on the biogenic substrates, and thus the urgency of developing a favorable amount of BTE degraders was reduced.     

Biodegradation of trichloroethylene and toluene by indigenous microbial populations in vadose sediments

The unsaturated subsurface (vadose zone) receives significant amounts of hazardous chemicals, yet little is known about its microbial communities and their capacity to biodegrade pollutants. Trichloroethylene (TCE) biodegradation occurs readily in surface soils; however, the process usually requires enzyme induction by aromatic compounds, methane, or other cosubstrates. The aerobic biodegradation of toluene and TCE by indigenous microbial populations was measured in samples collected from the vadose zone at unpolluted and gasoline-contaminated sites. Incubation at field moisture levels showed little activity on either TCE or toluene, so samples were tested in soil suspensions. No degradation occurred in samples suspended in water or phosphate buffer solution; however, both toluene and TCE were degraded in samples suspended in mineral salts medium. TCE degradation depended on toluene degradation, and little loss occurred under sterile conditions. Studies with specific nutrients showed that addition of ammonium sulfate was essential for degradation, and addition of other mineral nutrients further enhanced the rate. Additional studies with vadose sediments amended with nutrients showed similar trends to those observed in sediment suspensions. Initial rates of biodegradation in suspensions were faster in uncontaminated samples than in gasolinecontaminated samples, but the same percentages of chemicals were degraded. Biodegradation was slower and less extensive in shallower samples than deeper samples from the uncontaminated site. Two toluene-degrading organisms isolated from a gasoline-contaminated sample were identified as Corynebacterium variabilis SVB74 and Acinetobacter radioresistens SVB65. Inoculation with 10⁶ cells of C. variabilis ml^–1 of soil solution did not enhance the rate of degradation above that of the indigenous population. These results indicate that mineral nutrients limited the rate of TCE and toluene degradation by indigenous populations and that no additional benefit was derived from inoculation with a toluene-degrading bacterial strain. Correspondence to: K.M. Scow     

Biodegradation of trichloroethylene and toluene by indigenous microbial populations in soil.

The biodegradation of trichloroethylene (TCE) and toluene, incubated separately and in combination, by indigenous microbial populations was measured in three unsaturated soils incubated under aerobic conditions. Sorption and desorption of TCE (0.1 to 10 micrograms ml-1) and toluene (1.0 to 20 micrograms ml-1) were measured in two soils and followed a reversible linear isotherm. At a concentration of 1 micrograms ml-1, TCE was not degraded in the absence of toluene in any of the soils. In combination, both 1 microgram of TCE ml-1 and 20 micrograms of toluene ml-1 were degraded simultaneously after a lag period of approximately 60 to 80 h, and the period of degradation lasted from 70 to 90 h. Usually 60 to 75% of the initial 1 microgram of TCE ml-1 was degraded, whereas 100% of the toluene disappeared. A second addition of 20 micrograms of toluene ml-1 to a flask with residual TCE resulted in another 10 to 20% removal of the chemical. Initial rates of degradation of toluene and TCE were similar at 32, 25, and 18 degrees C; however, the lag period increased with decreasing temperature. There was little difference in degradation of toluene and TCE at soil moisture contents of 16, 25, and 30%, whereas there was no detectable degradation at 5 and 2.5% moisture. The addition of phenol, but not benzoate, stimulated the degradation of TCE in Rindge and Yolo silt loam soils, methanol and ethylene slightly stimulated TCE degradation in Rindge soil, glucose had no effect in either soil, and dissolved organic carbon extracted from soil strongly sorbed TCE but did not affect its rate of biodegradation.     

Biodegradation of phenanthrene by the indigenous microbial biomass in a zinc amended soil

AIMS: To study the effect of zinc on the biodegradation of phenanthrene by the microbial biomass in soil. METHODS AND RESULTS: Uncontaminated soil was amended with zinc and phenanthrene as single or co-contaminants, and microbial metabolic activity was measured using an intracellular dehydrogenase enzyme bioassay over 37 days. Contaminants were amended at optimum, action and double the action level specified in 'The New Dutch List' (Ministry of Housing, Spatial Planning and Environment, the Netherlands, 2000). Microbial activity in soils with zinc or phenanthrene alone indicated the presence of tolerant, albeit inhibited soil micro-organisms. A zinc concentration at the optimum level of 140 mg kg(-1) in the co-contaminated soil (phenanthrene at 40 mg kg(-1)) resulted in marginal stimulation of the rate of phenanthrene biodegradation. However, Zn2+ concentrations at the action and double the action level of zinc (720 and 1440 mg kg(-1)) inhibited phenanthrene degradation. CONCLUSIONS: Biodegradation of phenanthrene in soils co-contaminated with zinc at concentrations above the action value is impeded. SIGNIFICANCE AND IMPACT OF THE STUDY: Bioremediation efforts to remove polycyclic aromatic hydrocarbon in zinc co-contaminated soils are likely to be constrained.     

Impact of an indigenous microbial enhanced oil recovery field trial on microbial community structure in a high pour-point oil reservoir

Based on preliminary investigation of microbial populations in a high pour-point oil reservoir, an indigenous microbial enhanced oil recovery (MEOR) field trial was carried out. The purpose of the study is to reveal the impact of the indigenous MEOR process on microbial community structure in the oil reservoir using 16Sr DNA clone library technique. The detailed monitoring results showed significant response of microbial communities during the field trial and large discrepancies of stimulated microorganisms in the laboratory and in the natural oil reservoir. More specifically, after nutrients injection, the original dominant populations of Petrobacter and Alishewanella in the production wells almost disappeared. The expected desirable population of Pseudomonas aeruginosa, determined by enrichment experiments in laboratory, was stimulated successfully in two wells of the five monitored wells. Unexpectedly, another potential population of Pseudomonas pseudoalcaligenes which were not detected in the enrichment culture in laboratory was stimulated in the other three monitored production wells. In this study, monitoring of microbial community displayed a comprehensive alteration of microbial populations during the field trial to remedy the deficiency of culture-dependent monitoring methods. The results would help to develop and apply more MEOR processes.     

Biodegradation of solvents in varnish waste water of an airscrubber from an automobile varnish shop

The scrubber liquid of an automobile varnish shop, where varnish particle, pigments and resins are washed out, contains microorganisms, that are able to degrade the main compounds of the solvents in the waste gases, i.e. n-butanol, ethylbenzene and structural isomers of xylene, ethyltoluene and trimethylbenzene. The concentration of the compounds in the varnish waste water to be purified oscillated between 1000 and 1 ppm. The specific degradation efficiency could be increased 4 to 12 times by improvement in oxygen supply.A further enhancement was achieved by continuous addition of organic solvents in the gas phase. During the cultivation the number of different microorganisms of the mixed culture in the scrubber liquid was reduced. The biological system was also very stable and not sensitive to fluctuation in the solvent supply. Differences in the degradation capacity of structural isomers take place at the same concentration of aromatic hydrocarbons. The elimination of 2- or 4-ethyltoluene and 1,3,5- or 1,2,3-trimethylbenzene were 10 to 25% lower than 3-ethyltoluene or 1,2,4-trimethylbenzene.     

Effects of a bacterial consortium from acid mine drainage on cadmium phytoextraction and indigenous soil microbial community

Plant and Soil - A major concern in developing microbially-assisted phytoextraction (MAP) is that the effects of introduced microbes on indigenous soil microbial community are profound and...     

Spatial and temporal changes in the microbial community in an anaerobic swine waste treatment lagoon

Microorganisms are central to both the beneficial (organic degradation, nutrient removal, biogas production) and detrimental (odor production, pathogen contamination) effects of swine waste storage systems. In this study, both quantitative (real-time polymerase chain reaction) and qualitative (denaturing gradient gel electrophoresis, cloning, sequence analysis) molecular analyses were used to track spatial and temporal changes in the microbial community of swine slurry from a 0.4 ha anaerobic lagoon. The lagoon, located in a region of western Kentucky which has a humid, subtropical environment, was sampled on a monthly basis (n = 10) over a period of one year at four different depths (top, 51 cm from the top, 152 cm from the top, and bottom >198 cm). The concentration and diversity of Bacteroides sp. was seasonal (up to 90% decrease between March and June). Hespellia sp. and other clostridial species, on the other hand, were endemic in the slurry (concentrations up to 1.0 × 10⁷ cells mL^?1 slurry) regardless of time of the year or lagoon depth. Results suggest that there were seasonal effects on the microbial community in the swine lagoon, while the effect of depth was not as pronounced. Seasonal changes in the microbial community in stored wastes may be (directly or indirectly) correlated with changes in malodor emissions from lagoons.     

Methanogenic sarcina from an anaerobic microbial community degrading p-toluene sulfonate

The methanogenic strain MM isolated from an anaerobic microbial community degrading p-toluene sulfonate showed optimal values of temperature and pH for growth equal to 37 degrees C and 6.3-6.9, respectively. The doubling times of the isolate grown on methanol, acetate, and methylamines under the optimal conditions were 8.8, 19.1, and 10.3-28.1 h, respectively. The growth of strain MM was observed only when the cultivation medium contained casamino acids or p-toluene sulfonate. The G + C content of the DNA of the isolate was 40.3 mol%. This, together with DNA-DNA hybridization data, allowed the new isolate to be identified as a strain of the species Methanosarcina mazei. The new isolate differed from the known representatives of this species in that it was resistant to alkylbenzene sulfonates and able to demethylate p-toluene sulfonate when grown on acetate.     

Detoxification of 2,4,6-trichlorophenol by an indigenous bacterial community

An indigenous bacterial community capable of degrading 2,4,6-trichlorophenol (TCP) as the sole carbon source was isolated from the Riachuelo, a polluted river in Buenos Aires. The community consists of three gram-negative bacterial, non-fermentative strains, two of them was identified as belonging to genus Pseudomonas and the other to genus Stenotrophomonas. None of the individual strains were capable of degrading TCP as the sole carbon source. Aerobic biodegradation assays were performed using a 2-l microfermentor at 28 °C with agitation. Biodegradation was evaluated by spectrophotometry, chloride release, gas chromatography and microbial growth. Detoxification was evaluated by using Vibrio fischeri, Pseudokirchneriella subcapitata and Daphnia magna as test organisms. The indigenous bacterial community degrades 100 mg l^?1 of TCP in 27 h. The absence of metabolites and toxicity was proved at the end of the process. The influence of the initial concentration of compound, pH, cell inoculum, presence of other substrates and toxic related compounds in the biodegradation process was assayed. Also the application for polluted water was studied. The promising behavior of the bacterial community under the different conditions assayed allows us to suggest its possible use in remediation processes.     

Microbial characterization and hydrocarbon biodegradation potential of natural bilge waste microflora

Shipping operations produce oily wastes that must be managed properly to avoid environmental pollution. The aim of this study was to characterize microorganisms occurring in ship bilge wastes placed in open lagoons and, particularly, to assess their potential to degrade polycyclic aromatic hydrocarbons (PAHs). A first-order kinetic was suitable for describing hydrocarbon biodegradation after 17 days of treatment. The calculated rate constants were 0.0668 and 0.0513 day^–1 with a corresponding half-life of 10.3 and 13.5 days for the aliphatic and aromatic hydrocarbon fractions, respectively. At day 17, PAH removal percentages were: acenaphtylene 100, fluorene 95.2, phenanthrene 93.6, anthracene 70.3, and pyrene 71.5. Methyl phenanthrene removals were lower than that of their parent compound (3-methyl phenanthrene 83.6, 2-methyl phenanthrene 80.8, 1-methyl phenanthrene 77.3, 9-methyl phenanthrene 75.1, and 2,7-dimethyl phenanthrene 76.6). Neither pure cultures nor the microbial community from these wastes showed extracellular biosurfactant production suggesting that the addition of an exogenously produced biosurfactant may be important in enhancing hydrocarbon bioavailability and biodegradation. DNA analysis of bilge waste samples revealed a ubiquitous distribution of the nahAc genotype in the dump pools. Although almost all of the isolates grew on naphthalene as sole carbon source, only some of them yielded nahAc amplification under the experimental conditions used. The variety of PAHs in bilge wastes could support bacteria with multiple degradation pathways and a diversity of catabolic genes divergent from the classical nah-like type.     

Process stability and microbial community structure in anaerobic hydrogen-producing microflora from food waste containing kimchi

Hydrogen production by the dark fermentation of food wastes is an economic and environmentally friendly technology to produce the clean energy source as well as to treat the problematic wastes. However, the long-term operations of the continuous anaerobic reactor for fermentative hydrogen production were frequently unstable. In this study, the structure of microbial community within the anaerobic reactor during unstable hydrogen production was examined by denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) techniques. The changes in microbial community from H(2)-producing Clostridium spp. to lactic acid-producing Lactobacillus spp. were well coincident with the unexpected process failures and the changes of metabolites concentrations in the effluent of the anaerobic reactor. As the rate of hydrogen production decreased, effluent lactic acid concentration increased. Low rate of hydrogen production and changes in microbial community were related to the 'kimchi' content and storage temperature of food waste feed solution. After low temperature control of the storage tank of the feed solution, any significant change in microbial community within the anaerobic reactor did not occur and the hydrogen production was very stably maintained for a long time.     

Biodegradation of p-nitrophenol in an aqueous waste stream by immobilized bacteria

Microbiological analyses of activated sludge reactors after repeated exposure to 100 mg of p-nitrophenol (PNP) per liter resulted in the isolation of three Pseudomonas species able to utilize PNP as a sole source of carbon and energy. Cell suspensions of the three Pseudomonas sp., designated PNP1, PNP2, and PNP3, mineralized 70, 60, and 45% of a 70-mg/liter dose of PNP in 24, 48, and 96 h, respectively. Mass-balance analyses of PNP residues for all three cultures showed that undegraded PNP was less than 1% (less than 50 micrograms); volatile metabolites, less than 1%; cell residues, 8.4 to 14.9%; and water-soluble metabolites, 1.2 to 6.7%. A mixed culture of all three PNP-degrading Pseudomonas sp. was immobilized by adsorption onto diatomaceous earth biocarrier in a 1.75-liter Plexiglas column. The column was aerated and exposed to a synthetic waste stream containing 629 to 2,513 mg of PNP per liter at flow rates of 2 to 15 ml/min. Chemical loading studies showed that the threshold concentration for acute toxicity of PNP to the immobilized bacteria was 2,100 to 2,500 mg/liter. Further studies at PNP concentrations of 1,200 to 1,800 mg/liter showed that greater than 99 and 91 to 99% removal of PNP was achieved by immobilized bacteria at flow rates of 10 and 12 ml/min, respectively. These values represent hydraulic retention times of 48 to 58 min and PNP removal rates of 0.99 to 1.1 mg/h per g of biocarrier at 25 degrees C under optimal conditions. This study shows the successful use of immobilized bacteria technology to remove high concentrations of PNP from aqueous waste streams.     

Biodegradation of organic waste gas pollutants with bacterial suspensions

Summary A model system is used to study the reaction engineering fundamentals of the biological waste gas scrubbing process, and some kinetic measurements of the biodegradation of pollutants dissolved in the absorbent are presented.     

Biodegradation of phenanthrene by indigenous microorganisms in soils from Livingstone Island, Antarctica

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) in soils has been linked to history of exposure to PAHs and prevailing environmental conditions. This work assessed the capacity of indigenous microorganisms in soils collected in Livingstone Island (South Shetlands Islands, Antarctica) with no history of pollution (∑PAHs: 0.14-1.47?ng?g(-1) dw) to degrade (14) C-phenanhthrene at 4, 12 and 22?°C. The study provides evidence of the presence of phenanthrene-degrading microorganisms in all studied soils. Generally, the percentage of (14) C-phenanhthrene mineralized increased with increasing temperature. The highest extent of (14) C-phenanhthrene mineralization (47.93%) was observed in the slurried system at 22?°C. This work supports findings of the presence of PAH-degrading microorganisms in uncontaminated soils and suggests the case is the same for uncontaminated Antarctic remote soils.     

Characterization of cytochrome 579, an unusual cytochrome isolated from an iron-oxidizing microbial community

A novel, soluble cytochrome with an unusual visible spectral signature at 579 nm (Cyt(579)) has been characterized after isolation from several different microbial biofilms collected in an extremely acidic ecosystem. Previous proteogenomic studies of an Fe(II)-oxidizing community indicated that this abundant red cytochrome could be extracted from the biofilms with dilute sulfuric acid. Here, we found that the Fe(II)-dependent reduction of Cyt(579) was thermodynamically favorable at a pH of >3, raising the possibility that Cyt(579) acts as an accessory protein for electron transfer. The results of transmission electron microscopy of immunogold-labeled biofilm indicated that Cyt(579) is localized near the bacterial cell surface, consistent with periplasmic localization. The results of further protein analysis of Cyt(579), using preparative chromatofocusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, revealed three forms of the protein that correspond to different N-terminal truncations of the amino acid sequence. The results of intact-protein analysis corroborated the posttranslational modifications of these forms and identified a genomically uncharacterized Cyt(579) variant. Homology modeling was used to predict the overall cytochrome structure and heme binding site; the positions of nine amino acid substitutions found in three Cyt(579) variants all map to the surface of the protein and away from the heme group. Based on this detailed characterization of Cyt(579), we propose that Cyt(579) acts as an electron transfer protein, shuttling electrons derived from Fe(II) oxidation to support critical metabolic functions in the acidophilic microbial community.     

Critical evaluation of solid waste sample processing for DNA-based microbial community analysis

Landfills represent a unique microbial ecosystem and play a significant role in global biogeochemical processes. The study of complex ecosystems such as landfills using DNA-based techniques can be advantageous since they allow for analysis of uncultured organisms and offer higher resolution in measuring demographic and metabolic (functional) diversity. However, sample acquisition and processing from refuse is challenging due to material heterogeneity. Decomposed refuse was used to evaluate the effect of seven sample processing methods on Bacteria and Archaea community structure using T-RFLP. Bias was assessed using measured richness and by comparing community structure using multi-dimensional scaling (MDS). Generally, direct methods were found to be most biased while indirect methods (i.e., removal of cellular material from the refuse matrix before DNA extraction) were least biased. An indirect method using PO₄ buffer gave consistently high bacterial and archaeal richness and also resulted in 28 and 34% recovery of R. albus and M. formicicum spiked into refuse, respectively. However, the highest recovery of less abundant T-RFs was achieved using multiple processing methods. Results indicate differences in measured T-RF diversity from studies of landfill ecosystems could be caused by methodological (i.e., processing method) variation rather than refuse heterogeneity or true divergence in community structure.