[Microbiological degradation of mixed hydrocarbons in soil samples from a used-oil contaminated site].
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Bacteriological Techniques↗
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Selection of trichloroethene (TCE) degrading bacteria that resist inactivation by TCE.
Two isoprene (2-methyl-1,3-butadiene) utilizing bacteria, Alcaligenes denitrificans ssp. xylosoxidans JE 75 and Rhodococcus erythropolis JE 77, were identified as highly efficient cooxidizers of TCE, cis- and trans-dichloroethene, 1,1-dichloroethene and vinyl-chloride. Isoprene grown cells eliminate chloride from TCE in stoichiometric amounts and tolerate high concentrations of TCE.
Alcaligenes↗
Toxicity assessment of a former manufactured gas plant.
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Biological Assay↗
Isolation and characterization of psychotrophic bacteria from oil-reservoir water and oil sands.
Four psychrotrophic strains, which grew at 4 degrees C but not at 37 degrees C, were isolated from Japanese oil-reservoir water (strains SIB1, SIC1, SIS1) and Canadian oil sands (strain CAB1). Strains SIB1, SIS1, and CAB1 had a maximum growth rate at 20 degrees C and grew to the highest cell densities at the cultivation temperature of 0-4 degrees C. Strain SIS1 was capable of growing even at -5 degrees C. The growth profile of strain SIC1 was rather similar to that of a mesophilic bacterium. Strains SIB1, SIC1, and SIS1 were identified as members of the genus Shewanella, and strain CAB1 was a member of the genus Arthrobacter. All these strains exhibited weak degradation ability against catechol, a hydroxylated aromatic hydrocarbon, and tributyrin. These strains are expected to be of potential use in the in situ bioremediation technology of hazardous hydrocarbons and esters under low-temperature conditions.
Aerobiosis↗
Transformation of 2,4,6-trinitrotoluene (TNT) by Raoultella terrigena.
Manufacture of nitroorganic explosives generates toxic wastes leading to contamination of soils and waters, especially groundwater. For that reason bacteria living in environments highly contaminated with 2,4,6-trinitrotoluene (TNT) and other nitroorganic compounds were investigated for their capacity for TNT degradation. One isolate, Raoultella terrigena strain HB, removed TNT at concentrations between 10 and 100 mg l(-1) completely from culture supernatants under optimum aerobic conditions within several hours. Only low concentrations of nutrient supplements were needed for the cometabolic transformation process. Radioactivity measurements with ring-labelled (14)C-TNT detected about 10-20% of the initial radioactivity in the culture supernatant and the residual 80-90% as water-insoluble organic compounds in the cellular pellet. HPLC analysis identified aminodinitrotoluenes (2-ADNT, 4-ADNT) and diaminonitrotoluenes (2,4-DANT) as the metabolites which remained soluble in the culture medium and azoxy-dimers as the main products in the cell extracts. Hence, the new isolate could be useful for the removal of TNT from contaminated waters.
Aerobiosis↗
Contamination levels and preliminary assessment of the technical feasibility of employing natural attenuation in 5 priority areas of Presidente Bernardes Refinery in Cubatão, São Paulo, Brazil.
Five priority areas of potential impact by contaminants (API) were investigated at the Presidente Bernardes Refinery in Cubatão, São Paulo, Brazil with the following aims: (i) to identify both organic and inorganic contaminants present in soil and groundwater; (ii) to define the environmental conditions relevant for microbial activity at the site and (iii) to evaluate the feasibility of employing natural attenuation for treatment of the hydrocarbon contamination. One area (API 1) was an uncontrolled landfill, where waste materials from the refinery were deposited between 1954 and 1986, and four areas (API 4, 5, 7 and 11) were located in the operational section of the refinery. Soil contamination by regulated BTEX compounds (benzene, toluene, total xylenes) was restricted to two samples from API 1. Nonregulated ethylbenzene was detected in one soil sample from API 4, one from API 5 and two from API 1. No soil contained regulated PAH above threshold levels. Several nonregulated PAHs were found in 6 soil samples from API 1, 3 soil samples from API 4 and 1 soil sample from API 5. Site soils contained very high aluminium concentrations, but metal contamination was restricted to one soil sample from API 1, which contained nickel above threshold limits. BTEX contamination of groundwater was due mostly to benzene. Of the 17 PAH molecules tested, only naphthalene and 2-methylnaphthalene occurred in groundwater. The sum of total BTEX and total PAH exceeded 200 microg/L in only a few monitoring wells in API 4, 5 and 11 and was always below 2.640 microg/L. Be, Cd, Cr, Cu, Hg, Ni, Se, Ag, Tl and Zn were not detected in groundwater, which was contaminated in a few locations by aluminium (mostly below 1 mg/L), lead (<0.066 mg/L) and arsenic (<0.056 mg/L). S, K, Ca, Mg and Fe were present in groundwater in excess of physiological requirements for microbial growth, but low concentrations of N and P could become growth limiting. However, BTEX were efficiently degraded in saturated and unsaturated zone microcosms and nutrient amendments did not stimulate biodegradation rates measurably. The inorganic carbon pool in groundwater was up to one order of magnitude larger than the organic carbon pool. Total inorganic carbon (TIC) in API groundwater exceeded TIC of clean groundwater by factors of 2 (API 4), 6 (API 5, 7 and 11) or 10 (API 1). Most of the inorganic carbon incorporated into groundwater beneath the refinery originated from biodegradation in the unsaturated soil, which contained a microbiota (10(6) cells/g on average) capable of growth with most of the pure (benzene, toluene, ethylbenzene and xylene) and mixed hydrocarbons tested (diesel oil, gasoline, naphtha, condensate, aromatic residue and fuel oil). A viscous hydrocarbon paste uncovered in API 1 was insoluble in water but dissolved in dichloromethane. Many organic components of this paste were biodegradable as evidenced by weight reduction of the hydrocarbon paste and by the growth of suspended and attached biomass in saturated zone microcosms, where the paste was the only carbon source. This study indicates that monitored natural attenuation may be a technically feasible and efficient means for plume control in API 1, 4 and 5, provided the plumes in API 4 and 5 are not expanding. This technique is not suitable for contaminant reduction in API 11.
Biodegradation, Environmental↗
Dynamics and influencing factors of heterotrophic bacterial utilization of acetate in constructed wetlands treating woodwaste leachate.
14C-acetate was used as a tracer of substrates for heterotrophic bacteria to investigate the dynamics and influencing factors of bacterial activity in surface flow wetlands treating woodwaste leachate. Epiphytic assimilation and mineralization of acetate increased considerably over three weeks of microbial colonization. No significant longitudinal variation in acetate uptake was found. The temporal variation of bacterial activity was neither correlated to water temperature nor acetic acid concentration. Mineralization percentage varied significantly with acetate uptake rate. Acetate uptake rates of bacterioplankton (408+/-258 microg L(-1) h(-1)), epiphyton (67.7+/-41.7 mg m(-2) h(-1)) and sedimentary bacteria (26.7+/-25.6 microg g(-1) h(-1)) were influenced by the concentrations of organic substrates and inorganic nutrients. Sedimentary bacteria contributed to the majority (55-73%) of the total heterotrophic acetate uptake, while epiphytic bacteria played only a minor role (1-3%). The acetate mineralization percentage of sediment (16%) was much lower than that of water (55%) and epiphyton (64%). Addition of ammonium nitrate fertilizer increased acetate uptake rates significantly, especially for sediment. No remarkable changes in mineralization percentage and the relative importance of water, sediment and epiphyton were found due to fertilization.
Acetates↗
Anaerobic degradation of toluene and o-xylene by a methanogenic consortium.
Toluene and o-xylene were completely mineralized to stoichiometric amounts of carbon dioxide, methane, and biomass by aquifer-derived microorganisms under strictly anaerobic conditions. The source of the inoculum was creosote-contaminated sediment from Pensacola, Fla. The adaptation periods before the onset of degradation were long (100 to 120 days for toluene degradation and 200 to 255 days for o-xylene). Successive transfers of the toluene- and o-xylene-degrading cultures remained active. Cell density in the cultures progressively increased over 2 to 3 years to stabilize at approximately 10(9) cells per ml. Degradation of toluene and o-xylene in stable mixed methanogenic cultures followed Monod kinetics, with inhibition noted at substrate concentrations above about 700 microM for o-xylene and 1,800 microM for toluene. The cultures degraded toluene or o-xylene but did not degrade m-xylene, p-xylene, benzene, ethylbenzene, or naphthalene. The degradative activity was retained after pasteurization or after starvation for 1 year. Degradation of toluene and o-xylene was inhibited by the alternate electron acceptors oxygen, nitrate, and sulfate. Degradation was also inhibited by the addition of preferred substrates such as acetate, H2, propionate, methanol, acetone, glucose, amino acids, fatty acids, peptone, and yeast extract. These data suggest that the presence of natural organic substrates or contaminants may inhibit anaerobic degradation of pollutants such as toluene and o-xylene at contaminated sites.
Anaerobiosis↗
Microbial diversity in a hydrocarbon- and chlorinated-solvent-contaminated aquifer undergoing intrinsic bioremediation.
A culture-independent molecular phylogenetic approach was used to survey constituents of microbial communities associated with an aquifer contaminated with hydrocarbons (mainly jet fuel) and chlorinated solvents undergoing intrinsic bioremediation. Samples were obtained from three redox zones: methanogenic, methanogenic-sulfate reducing, and iron or sulfate reducing. Small-subunit rRNA genes were amplified directly from aquifer material DNA by PCR with universally conserved or Bacteria- or Archaea-specific primers and were cloned. A total of 812 clones were screened by restriction fragment length polymorphisms (RFLP), approximately 50% of which were unique. All RFLP types that occurred more than once in the libraries, as well as many of the unique types, were sequenced. A total of 104 (94 bacterial and 10 archaeal) sequence types were determined. Of the 94 bacterial sequence types, 10 have no phylogenetic association with known taxonomic divisions and are phylogenetically grouped in six novel division level groups (candidate divisions WS1 to WS6); 21 belong to four recently described candidate divisions with no cultivated representatives (OP5, OP8, OP10, and OP11); and 63 are phylogenetically associated with 10 well-recognized divisions. The physiology of two particularly abundant sequence types obtained from the methanogenic zone could be inferred from their phylogenetic association with groups of microorganisms with a consistent phenotype. One of these sequence types is associated with the genus Syntrophus; Syntrophus spp. produce energy from the anaerobic oxidation of organic acids, with the production of acetate and hydrogen. The organism represented by the other sequence type is closely related to Methanosaeta spp., which are known to be capable of energy generation only through aceticlastic methanogenesis. We hypothesize, therefore, that the terminal step of hydrocarbon degradation in the methanogenic zone of the aquifer is aceticlastic methanogenesis and that the microorganisms represented by these two sequence types occur in syntrophic association.
Bacteria↗
[The ecological problems of the health resort and recreational use of bodies of water].
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Communicable Diseases↗
Review of whole-organism bioassays: soil, freshwater sediment, and freshwater assessment in Canada.
Whole organism bioassays for the assessment of soil, freshwater sediment, and freshwater quality were evaluated for their application in the assessment and remediation of contaminated sites in Canada under the National Contaminated Sites Remediation Program. Using 3 essential and 12 desirable methodological criteria, bioassays were categorized as currently usable, prototype, or under development. Based on further considerations related to bioassay application, a battery of usable screening and definitive tests was recommended (with suggestions for augmentation) for each medium. Of the 18 bioassays reviewed for soil quality assessment, 6 were usable, 5 were prototypes, and 7 were under development. Battery screening and definitive tests included 14-day Eisenia andrei survival, 120-hr lettuce and radish seedling emergence, and 72-hr Selenastrum capricornutum growth inhibition. Augmentation with the following bioassays was recommended: soil/freshwater bacterial growth, arthropod reproduction, earthworm reproduction, and reproduction of other soil-dependent organisms. Of the 9 bioassays reviewed for freshwater sediment quality assessment, 1 was usable, 2 were prototypes, and 6 were under development. Three bioassays in the latter two groups were considered usable with the imminent completion of research underway. Screening tests selected included 10-day Chironomus tentans survival, 10-day Hyalella azteca survival, 10-day Hexagenia spp. survival, and 72-hr S. capricornutum growth inhibition. Definitive tests included screening tests, substituting 28-day H. azteca sexual maturation for 10-day survival. Augmentation with the following bioassays was recommended: sediment/freshwater bacterial test, 28-day Tubifex tubifex reproduction, and rooted aquatic plant growth. Of the 25 bioassays considered for freshwater quality assessment, 8 were usable, 7 were prototypes, and 10 were under development. Screening tests selected included 72-hr S. capricornutum growth inhibition; 48-hr Daphnia sp. survival, and 5- and 15-min Photobacterium phosphoreum bioluminescence. Definitive tests included first screening test, 7-day Ceriodaphnia dubia, 7-day fathead minnow larval survival, or 96-hr rainbow trout survival. Augmentation with the following bioassays was recommended: Brachionus calyciflorus 24-hr survival, 48-hr reproduction; freshwater bacterial growth; and aquatic vascular plant growth.
Animals↗
Concentrations of selected heavy metals in benthic diatoms and sediment in the Westerschelde estuary.
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Cadmium↗
Studies with natural and anthropogenic iodine isotopes: iodine distribution and cycling in the global environment.
Iodine is an important trace element in geological and biological processes. We summarize here recent results and new data of experiments and observations carried out to improve the understanding of concentration levels and behavior of natural and anthropogenic iodine nuclides in the global environment. The distribution of stable iodine in the Earth's crust was estimated using concentration data in a suite of representative samples and the influence of subduction on the marine iodine cycle was investigated using (129)I systematics on iodine-rich brines from Japan. The importance of microorganisms for the natural iodine cycle is shown in recent studies of iodine sorption on soil and of iodine volatilization from terrestrial and marine environments. Levels of anthropogenic (129)I were measured in samples collected around a spent fuel reprocessing plant in Japan.
Environmental Monitoring↗
Field and numerical analysis of in-situ air sparging: a case study.
An in-situ air sparging operation was used to remediate the sandy subsurface soils and shallow groundwater under a drum storage site near Chicago, IL, where either periodic or random spillage of a light non-aqueous phase liquid (LNAPL) occurred between 1980 and 1987. Both field measurements and model simulations using commercially available computer software suggested that microbial degradation was the most significant contributor to the removal of contaminant mass. Toluene, ethylbenzene and total xylenes (TEX), which were of major concern with regards to reaching clean-up criteria at the site, were observed to decline by 88% in concentration. Furthermore, up to 97% of the total mass removed through microbial degradation consisted of TEX. Of the total contaminant spill, up to 23% of initial organic chemical mass was removed through microbial degradation compared to less than 6% by physical stripping. Greater loss to microbial degradation is most likely attributed to the relatively low air injection rate used during the course of the air sparging remediation. Evaluation of air sparging at the site using model simulations supported this analysis by estimating 140 and 620 kg of total contaminant mass being removed through volatilization and biodegradation, respectively. An evaluation of several system design parameters using model simulations suggested that only the type of sparging operation (i.e. pulsed or continuous) was significant in terms of total contaminant removal time, while both the sparging operation and air injection rate were significant in terms of removal of a critical species, total xylenes.
Air↗
Reduction of hazardous levels of the agricultural application of nitrogen and phosphorus relative to toxic ground water and toxic levels in the soil.
This paper proposes the hypothesis that microbial life chemically reduces levels of nitrogen (N(2)) and phosphorus (P) that are toxic and threaten human health and safety. Bio-remediation uses microorganisms to decontaminate a polluted system, in situ, requiring a minimal amount of space and equipment. Data strongly suggest that bio-stimulation can assist one microbe to multiply up to one billion microorganisms in 24 hours. Biochemical literature postulates that microbial life chemically biodegrades nitrates by one of two methods: (1) assimilative reduction; or (2) dissimilative reduction, also known as denitrification. Assimilative reduction results in construction of microbial cell walls, cell membranes and various forms of amino acids. It is proposed that denitrification includes the venting-off of the excess amounts of N(2)not required by the soil or needed for additional microbial development. Nitrate reduction by way of denitrification is a functional part of anaerobic respiration. Alternatively, the denitrification process supports oxidative phosphorylation, a mechanism similar to aerobic respiration. Thus, denitrification and phosphorylation may be considered as forms of respiration.
Agriculture↗
Molecular analysis of Dehalococcoides 16S ribosomal DNA from chloroethene-contaminated sites throughout North America and Europe.
The environmental distribution of Dehalococcoides group organisms and their association with chloroethene-contaminated sites were examined. Samples from 24 chloroethene-dechlorinating sites scattered throughout North America and Europe were tested for the presence of members of the Dehalococcoides group by using a PCR assay developed to detect Dehalococcoides 16S rRNA gene (rDNA) sequences. Sequences identified by sequence analysis as sequences of members of the Dehalococcoides group were detected at 21 sites. Full dechlorination of chloroethenes to ethene occurred at these sites. Dehalococcoides sequences were not detected in samples from three sites at which partial dechlorination of chloroethenes occurred, where dechlorination appeared to stop at 1,2-cis-dichloroethene. Phylogenetic analysis of the 16S rDNA amplicons confirmed that Dehalococcoides sequences formed a unique 16S rDNA group. These 16S rDNA sequences were divided into three subgroups based on specific base substitution patterns in variable regions 2 and 6 of the Dehalococcoides 16S rDNA sequence. Analyses also demonstrated that specific base substitution patterns were signature patterns. The specific base substitutions distinguished the three sequence subgroups phylogenetically. These results demonstrated that members of the Dehalococcoides group are widely distributed in nature and can be found in a variety of geological formations and in different climatic zones. Furthermore, the association of these organisms with full dechlorination of chloroethenes suggests that they are promising candidates for engineered bioremediation and may be important contributors to natural attenuation of chloroethenes.
Bacteria, Anaerobic↗
[Microbiologic/hygienic evaluation of the risk potential of bacteria from soil and water sources in combination with biotechnologic risks of soil restoration].
Biological cleanup-processes removing contaminations from soil and ground-water can be carried out "ex-situ" or "in situ". The pollutants are mineralized (totally or partly) by microorganisms. The successful application of bioremediation techniques often leads to an increase of aerobic and facultatively anaerobic bacteria in soil and/or groundwater, a fact that triggered discussions about a potential biological risk originating from these microorganisms. In this study the identification results of 10,200 microorganisms isolated from 109 ground-water- and 152 soil-samples of different sites contaminated with mineral oil or chlorinated aromatics are evaluated with respect to a possible biological risks originating from these microorganisms. According to their biological risk potential, 19.2% (1958) of the isolated organisms can be grouped into risk-groups II, that means they they are able o cause human disease but it is unlikely that they are spread to the community and there is effective prophylaxis or treatment available. The majority of isolates (81.8%) are grouped into risk group I, meaning that it is very unlikely that they cause a human disease. All these microorganisms are either adsorbed to the soil-matrix or within a closed system of a water-treatment-plant, so that the risk of infection is very unlikely for healthy individuals (regarding possible pathways of infection). The risk potential in waste-water-treatment-plants is regarded to be significantly higher than in biological soil-cleanup processes, because of the specific origin of the bacteria and the possible formation of aerosols.
Bacteria, Aerobic↗
A simplified dehydrogenase enzyme assay in contaminated sediment using 2-(p-Iodophenyl)-3(p-nitrophenyl)-5-phenyl tetrazolium chloride.
2-(p-Iodophenyl)-3(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) accepts electrons from dehydrogenase enzymes and is reduced to a red-colored formazan (INTF), which can be quantified by colorimetric analysis. Use of previously published methods for this technique was unsuccessful due to background chemical reactions from high levels of polycyclic aromatic hydrocarbons (PAHs) and metals in the sediments. A modified method using acetonitrile extraction of the INTF was efficient and did not chemically reduce INT. This activity method is simple, quick, inexpensive and precise.
Aerobiosis↗