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Structure of microbial communities in activated sludge: potential implications for assessing the biodegradability of chemicals.

Various methods used to assess the biodegradability of chemicals often employ activated sludge as an inoculum since chemicals that ultimately enter the environment are often discharged through wastewater. Differences in the structure and function of activated sludge microbial communities that may complicate interpretation of biodegradation tests could arise from differences in wastewater composition, wastewater treatment plant (WWTP) operation, or manipulations done after collection of the activated sludge. In this study, various methods were used to characterize the structure of microbial communities found in freshly collected activated sludge from WWTPs in Japan, Europe, and the United States, as well as sludge that had been continuously fed either sewage or a glucose-peptone mixture for several weeks after collection. Comparisons of biomass levels, whole-community substrate utilization (determined using Biolog GN and GP plates), and phospholipid fatty acid (PLFA) profiles indicated there were both geographical and temporal differences among freshly collected activated sludge samples. Moreover, marked shifts in the structure of activated sludge microbial communities occurred upon continuous cultivation in the laboratory for 5 weeks using a glucose-peptone feed. These shifts were evident from whole-community substrate utilization and PLFA profiles as well as differences in the profiles of 16S rDNA genes from numerically dominant populations obtained by denaturing gradient gel electrophoresis and terminal restriction fragment analyses. Further studies are needed to better define the variability within and between activated sludge from wastewater treatment plants and laboratory reactors and to assess the impact of such differences on the outcome of biodegradability tests.

Bacteria↗

Biodegradability of new engineered fuels compared to conventional petroleum fuels and alternative fuels in current use.

Concern with environmental issues such as global climate change has stimulated research into the development of more environmentally friendly technologies and energy sources. One critical area of our economy is liquid transportation fuels. This article presents the results of the biodegradability potential of newly developed engineered fuels and compares the results to the biodegradability of conventional fuels and alternative fuels in current use. Biodegradability potential was determined under both aerobic and anaerobic conditions. Fuels that have a high degree of components derived from renewable sources proved to have a higher degradability potential than those composed of petroleum components.

Biodegradation, Environmental↗

Dynamic response of naphthalene biodegradation in a continuous flow soil slurry reactor.

Periodic perturbations were used to evaluate the system stability and robustness of naphthalene biodegradation in a continuous flow stirred tank reactor (CSTR) containing a soil slurry. The experimental design involved perturbing the test system using a sinusoidal input either of naphthalene or non-naphthalene organic carbon at different frequencies during steady state operation of the reactors. The response of the test system was determined by using time series off-gas analysis for naphthalene liquid phase concentration and degradation, total viable cell counts, and gene probe analysis of naphthalene degradative genotype, and by batch mineralization assays. Naphthalene biodegradation rates were very high throughout the experimental run (95 to greater than 99% removed) resulting in very low or undetectable levels of naphthalene in the off-gas and reactor effluent. Attempts to reduce the rate of naphthalene biotransformation by either reducing the reactor temperature from 20 degrees C to 10 degrees C or the dissolved oxygen level (greater than 1 mg/L) were unsuccessful. Significant naphthalene biodegradation was observed at 4 degrees C. While variable, the microbial community as measured by population densities was not significantly affected by temperature changes. In terms of naphthalene biotransformation, the system was able to adapt readily to all perturbations in the reactor.

Biodegradation, Environmental↗

Biodegradation of atrazine in surface soils and subsurface sediments collected from an agricultural research farm.

The purpose of the present study was to assess atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) mineralization by indigenous microbial communities and to investigate constraints associated with atrazine biodegradation in environmental samples collected from surface soil and subsurface zones at an agricultural site in Ohio. Atrazine mineralization in soil and sediment samples was monitored as 14CO2 evolution in biometers which were amended with 14C-labeled atrazine. Variables of interest were the position of the label ([U-14C-ring]-atrazine and [2-14C-ethyl]-atrazine), incubation temperature (25 degrees C and 10 degrees C), inoculation with a previously characterized atrazine-mineralizing bacterial isolate (M91-3), and the effect of sterilization prior to inoculation. In uninoculated biometers, mineralization rate constants declined with increasing sample depth. First-order mineralization rate constants were somewhat lower for [2-14C-ethyl]-atrazine when compared to those of [U-14C-ring]-atrazine. Moreover, the total amount of 14CO2 released was less with [2-14C-ethyl]-atrazine. Mineralization at 10 degrees C was slow and linear. In inoculated biometers, less 14CO2 was released in [2-14C-ethyl]-atrazine experiments as compared with [U-14C-ring]-atrazine probably as a result of assimilatory incorporation of 14C into biomass. The mineralization rate constants (k) and overall extents of mineralization (Pmax) were higher in biometers that were not sterilized prior to inoculation, suggesting that the native microbial populations in the sediments were contributing to the overall release of 14CO2 from [U-14C-ring]-atrazine and [2-14C-ethyl]-atrazine. A positive correlation between k and aqueous phase atrazine concentrations (Ceq) in the biometers was observed at 25 degrees C, suggesting that sorption of atrazine influenced mineralization rates. The sorption effect on atrazine mineralization was greatly diminished at 10 degrees C. It was concluded that sorption can limit biodegradation rates of weakly-sorbing solutes at high solid-to-solution ratios and at ambient surface temperatures if an active degrading population is present. Under vadose zone and subsurface aquifer conditions, however, low temperatures and the lack of degrading organisms are likely to be primary factors limiting the biodegradation of atrazine.

Adsorption↗

Extraction and biodegradation of a toxic volatile organic compound (1,2-dichloroethane) from waste-water in a membrane bioreactor.

An extractive membrane bioreactor has been used to treat a synthetic waste-water containing a toxic volatile organic compound, 1,2-dichloroethane (DCE). Biofilms growing on the surface of the membrane tubes biodegrade DCE while avoiding direct contact between the DCE and the aerating gas. This reduces air stripping by more than an order of magnitude (from 30-35% of the DCE entering the system to less than 1%) relative to conventional aerated bioreactors. Over 99% removal of DCE from a waste-water containing 1600 mg l-1 of DCE was achieved at waste-water residence times of 0.75 h. Biodegradation was verified as the removal mechanism through measurements of CO2 and chloride ion evolution in the bioreactor. No DCE was detected in the biomedium over the operating period. The diffusion-reaction phenomena occurring in the biofilm have been described by a mathematical model, which provides calculated solutions that support the experimental results by predicting that all DCE is biodegraded within the biofilm. Experimentally, however, the rate of DCE degradation in the biofilm was found to be independent of O2 concentration, while the model predictions point to O2 being limiting.

Biodegradation, Environmental↗

Use of continuous-flow UV-induced mutation technique to enhance chlorinated organic biodegradation.

In this study, a continuous-flow UV-induced mutation (CUM) device and the CUM device coupled to a selector (CUMS) reactor were fabricated and tested for their ability to enhance the probability of obtaining populations capable of chlorinated organic biodegradation. A mixed culture of bacteria were used as the starting strain for both the CUM and CUMS processes. Populations were obtained from the CUM and CUMS systems capable of 4-chlorobenzoic acid, 2,4-dichlorobenzoic acid and chlorendic acid biodegradation. Non-UV irradiated population served as controls for the experiments and did not demonstrate chlorinated organic biodegradation over the test duration.

Bacteria↗

Protection of intestinal anastomoses by biodegradable intraluminal bypass tubes under the condition of general peritonitis: an experimental study on the CLP model in rats.

The purpose of this experimental study was to affirm the protective effect of biodegradable tubes for the intraluminal bypass procedure under the adverse condition of general peritonitis. General peritonitis was induced by means of the cecal ligation and puncture (CLP) model in the rat. The leakage rate in the control group (n = 20) without anastomotic protection was 70 percent (14/20). In three therapeutic groups, each consisting of 20 animals, the intestinal anastomoses were protected by an intraluminal bypass tube of different biodegradable biomaterials (collagen-II, BCL-002, and BCL-004). The best results were noted in the collagen-II and BCL-002 groups, where the leakage rates could be reduced to 10 percent. These highly significant results (P = 0.0001) prove the feasibility of biodegradable biomaterials for the intraluminal bypass procedure in the rat, even in cases with underlying peritonitis.

Anastomosis, Surgical↗

Lignin biodegradation by the ascomycete Chrysonilia sitophila.

The lignin biodegradation process has an important role in the carbon cycle of the biosphere. The study of this natural process has developed mainly with the use of basidiomycetes in laboratory investigations. This has been a logical approach since most of the microorganisms involved in lignocellulosic degradation belong to this class of fungi. However, other microorganisms such as ascomycetes and also some bacteria, are involved in the lignin decaying process. This work focuses on lignin biodegradation by a microorganism belonging to the ascomycete class, Chrysonilia sitophila. Lignin peroxidase production and characterization, mechanisms of lignin degradation (lignin model compounds and lignin in wood matrix) and biosynthesis of veratryl alcohol are outstanding. Applications of C. sitophila for effluent treatment, wood biodegradation and single-cell protein production are also discussed.

Ascomycota↗

Identification of alkylpolyoxyethylene glycols in the residues after biodegradation of their mixtures by CGC and previous derivatization.

This study deals with the identification of individual n-alkylpolyoxyethylene glycols among the products of oxyethylenation of n-nonyl- and n-decylalcohol and with the identification of these compounds in the reaction mixture after their biodegradation. The individual glycols were identified by their retention indices for the column temperature programmed linearly, by the correlation of the retention indices with the number of oxyethylene groups in the molecule and by increments of retention indices for the incoming oxyethylene group or methylene group. The biodegradation process of oxyethylenation products of reaction of one mole of n-nonylalcohol and n-decylalcohol, respectively, with three moles of ethylene oxide was studied. n-Alkylpolyoxyethylene glycols in oxyethylenation products or their residues isolated from the aqueous solution after their biodegradation processes were derivatized into acetates, trifluoroacetates and trimethylsilyl derivatives before carrying out their identification and determination.

Acetates↗

Biodegradable plastics from renewable sources.

Plastic waste disposal is a huge ecotechnological problem and one of the approaches to solving this problem is the development of biodegradable plastics. This review summarizes data on their use, biodegradability, commercial reliability and production from renewable resources. Some commercially successful biodegradable plastics are based on chemical synthesis (i.e. polyglycolic acid, polylactic acid, polycaprolactone, and polyvinyl alcohol). Others are products of microbial fermentations (i.e. polyesters and neutral polysaccharides) or are prepared from chemically modified natural products (e.g., starch, cellulose, chitin or soy protein).

Bacteria↗

Estimating biodegradative gene numbers at a JP-5 contaminated site using PCR.

We have utilized a most-probable-number polymerase chain reaction (MPN-PCR) procedure to estimate gene numbers and biodegradative potential at a jet fuel (JP-5) contaminated site undergoing the first phase of bioremediation. Nucleic acid analysis was used to determine whether a lack of genetic potential for bioremediation was responsible for low levels of oxygen utilization at the site. Total community DNA was extracted and analyzed by PCR for genes (nahAc,alkB, and xylE) known to be involved in the degradation of certain JP-5 constituents. Results indicate that significant aromatic biodegradative potential exists at the site and outlying areas not subjected to engineered remediation, suggesting that physical and/or chemical factors are inhibiting oxygen delivery. xylE and nahAc were often present in significant portions of the microbial community, whereas alkB was rarely detected. This study illustrates the utility of molecular techniques in evaluating biodegradative potential in the field during active bioremediation.

Base Sequence↗

Decolorization and biodegradation of dye wastewaters by a facultative-aerobic process.

BACKGROUND: Dye wastewater is one of the main pollution sources of water bodies in China. Conventional biological processes are relatively ineffective for color removal, the development of alternative treatment methods will become important. Our subjective was that of introducing a new biotreatment technology which combined a facultative biofilm reactor (FBR) with an aerobic reactor (AR) to treat a dye wastewater. The efficiencies of color and chemical oxygen demand (COD) removal and the mechanism of dye degradation were investigated. METHODS: The anthraquinone acid dye (acid blue BRLL) concentration, organic loading rate (OLR) and hydraulic retention time (HRT) were varied in the experiments to evaluate the treatment efficiency and process stability. The biodegradation products were detected by infrared (IR) and high performance liquid chromatography and mass spectrometry (HPLC-MS). RESULTS AND DISCUSSION: The results demonstrated that the facultative biofilm process was more effective for decolorization than the anaerobic stage of an anaerobic-aerobic process. Most color removal occurred in the facultative reaction (maximum to 88.5%) and the BOD (biochemical oxygen demand): COD of the FBR effluent increased by 82.2%, thus improving the biodegradability of dyes for further aerobic treatment. The dye concentration, OLR and HRT will be the factors affecting decolorization. Color removal efficiency falls as the influent dye concentration increases, but rises with increased HRT. The infrared and HPLC-MS analyses of the effluents of FBR and AR reveal that the dye parent compound was degraded in each reactor during the process. CONCLUSION: The Facultative-aerobic (F-A) system can effectively remove both color and COD from the dye wastewater. The FBR played an essential role in the process. The average overall color and COD in the system were removed by more than 93.9% and 97.1%, respectively, at an OLR of 1.1 kg COD m(-3) d(-1) and at the HRT of 18-20 hours in the FBR and 4-5 hours in the AR. The color removal mechanism in each reactor was not only a sort of biosorption on the floc materials, but even more an effect of biodegradation, especially in the facultative process. Recommendation and Outlook. In applying the F-A system to treat a dye wastewater, the control of facultative processes and the set up of appropriate operation conditions appear to be critical factors. Also, it is suggested a moderate COD loading rate and about a 24-hour HRT will favor the F-A system.

Bacteria, Aerobic↗

[Local foreign body reactions to biodegradable implants. A classification].

Biodegradable implants are increasingly used in orthopedic and trauma surgery. Many different implants consisting of different biodegradable polymers are currently available. Different factors contribute to the biocompatibility of these implants, and local foreign-body reactions remain a matter of concern. Therefore, it is mandatory to document and compare the tissue reactions caused by various biodegradable implants in experimental or clinical studies. We have developed a standardized system of classification based on our previous experimental and clinical observations. Foreign-body reactions are differentiated into osteolysis (0-0 to 0-4), extra-articular (EA-0 to EA-4) and intraarticular (IA-0 to A-4) soft-tissue reactions.

Animals↗

Analysis and toxicity of methomyl and ametryn after biodegradation.

The controlled biodegradation of ametryn and methomyl has been performed, in accordance with the OECD Zahn-Wellens/EMPA procedure, by use of an enriched mixture of activated sludge collected from three domestic waste-water-treatment plants (WWTP). During the process concentrations of ametryn and methomyl in the water samples were isolated by solid-phase extraction (SPE); recovery rates were 98.9 and 93.2 for methomyl and ametryn, respectively. Liquid chromatography-mass spectrometry (LC-MS) was used to determine final pesticide concentrations and for metabolite identification. The efficiency of aerobic biodegradation of ametryn and methomyl was evaluated by measuring both the decrease in the concentration of the pesticides and global properties such as the chemical oxygen demand (COD). The acute toxicity of ametryn and methomyl was evaluated by use of the ToxAlert100 biological test, which is based on inhibition of the bioluminescence of Vibrio fischeri. There was significant correlation between results from primary and ultimate biodegradation and those from determination of toxicity. Pesticide concentrations were always reduced to below the limit of detection in less than 17 days. High COD removal (90-96%) was achieved in 28 and 18 days for methomyl and ametryn, respectively.

Biodegradation, Environmental↗

Effects of fractions from biodegraded Alaska North Slope crude oil on embryonic inland silversides, Menidia beryllina.

Embryonic inland silversides, Meinida beryllina, were exposed to neutral, water-soluble fractions (WSFs) resulting from microbial degradation of artificially weathered Alaska North Slope (ANS) crude oil. Three individual microbes obtained from Prince William Sound, Alaska, and designated Phe#6 (enriched on phenanthrene), Hexaco#2 (enriched on the straight-chain alkane, hexacosane), and EI2V (grown by enrichment on Bushnell-Haas medium containing 0.2% pristane, a branched alkane) were used to individually biodegrade weathered ANS crude oil for 14 days in darkness in 20-L glass carboys containing nutrient enriched, sterilized 20% salinity sea water at 20 +/- 1 degrees C. Neutral WSFs resulting from biodegradation of ANS (lot 521) by each microbe were recovered and weighted. Neutral WSFs recovered were: 1.76 mg/L for Phe#6, 1.85 mg/L for Hexaco#2, and 13.02 mg/L for the EI2V microbe. Embryo toxicity and teratogenicity tests revealed that exposure of embryos to the WSFs from the EI2V incubation (with a total recovered neutral fraction approximately seven times greater than the Phe#6 and Hexaco#2 incubations) resulted in the most severe responses in craniofacial, cardiovascular, and skeletal organ systems. The total neutral WSFs recovered from the EI2V biodegradation of weathered ANS 521 were subfractionated into saturated (eluted with hexane), aromatic (eluted with CH2Cl2), polar (eluted with ethyl ether), and recombined (saturated + aromatic + polar) fractions. Developing fish embryos were then exposed to each subfraction and the recombined subfractions. The polar subfraction and recombined subfractions proved to be the most embryo toxic and teratogenic. They resulted in statistically significant (p < or = 0.05) responses (compared to controls) for craniofacial, cardiovascular, skeletal, and total severity effects in one or both tests with these subfractions.

Animals↗

Biodegradation and ecotoxicity of HFCs and HCFCs.

Hydrofluorocarbons (HFCs) and hydrochlorofluorocarbons (HCFCs) are used or developed as substitutes for fully halogenated chlorofluorocarbons. Based on the results of closed-bottle tests, the biodegradation of HFC-32, HCFC-123, HCFC-124, HFC-125, HFC-134a, HCFC-141b, HCFC-225ca, and HCFC-225cb was less than 60% after 28 days and therefore these compounds are considered not readily biodegradable. Standard acute toxicity tests with HCFC-123, HCFC-141b, and HCFC-225ca using algae, water fleas, and fish revealed EC50 values in the range of 17-126 mg/L. EC50 values of HFC-134a ranged between 450-980 mg/L. Fish studies with HCFC-141b and HCFC-225ca revealed bioaccumulation factors of <3 and 15-64, respectively. A study with plants revealed no effect of HCFC-141b on seed germination and growth of wheat (Triticum aestivum), radish (Raphanus sativus), and cress (Lepidium sativum). In conclusion, HFCs and HCFCs are not very toxic to aquatic organisms and terrestrial plants. No evidence for any aerobic biodegradation for most of the HFCs and HCFCs was found.

Animals↗

Geochemical and physiological evidence for mixed aerobic and anaerobic field biodegradation of coal tar waste by subsurface microbial communities.

We used geochemical analyses of groundwater and laboratory-incubated microcosms to investigate the physiological responses of naturally occurring microorganisms to coal-tar-waste constituents in a contaminated aquifer. Waters were sampled from wells along a natural hydrologic gradient extending from uncontaminated (1 well) into contaminated (3 wells) zones. Groundwater analyses determined the concentrations of carbon and energy sources (pollutants or total organic carbon), final electron acceptors (oxygen, nitrate, sulfate), and metabolic byproducts (dissolved inorganic carbon [DIC], alkalinity, methane, ferrous iron, sulfide, Mn2+). In the contaminated zone of the study site, concentrations of methane, hydrogen, alkalinity, and DIC were enhanced, while dissolved oxygen and nitrate were depleted. Field-initiated biodegradation assays using headspace-free serum bottle microcosms filled with groundwater examined metabolism of the ambient organic contaminants (naphthalene, 2-methylnaphthalene, benzothiophene, and indene) by the native microbial communities. Unamended microcosms from the contaminated zone demonstrated the simultaneous degradation of several coal-tar-waste constituents at the in situ temperature (10 degrees C). Lag phases prior to the onset of biodegradation indicated the prevalence of both aerobic and anaerobic conditions in situ. Electron acceptor-amended microcosms from the most contaminated well waters demonstrated only aerobic naphthalene degradation. Collectively, the geochemical and microbial evidence show that biodegradation of coal-tar-waste constituents occurs via both aerobic and anaerobic terminal electron accepting processes at this site.

Bacteria, Aerobic↗

Effects of randomly methylated-beta-cyclodextrins (RAMEB) on the bioavailability and aerobic biodegradation of polychlorinated biphenyls in three pristine soils spiked with a transformer oil.

The low bioavailability of polychlorinated biphenyls (PCBs) in soils often results in their slow and partial aerobic biodegradation. The process can be enhanced by supplementing soils with cyclodextrins. However, pure cyclodextrins are expensive and we have therefore explored the use of a less costly technical grade mixture of randomly methylated-beta-cyclodextrins (RAMEB). RAMEB was tested at 0, 1, 3 and 5% (w/w) in the aerobic bioremediation and detoxification of a loamy-, a humic- and a sandy-soil, each artificially contaminated with a PCB-containing transformer oil (added PCBs: about 450 or 700 mg/kg), inoculated with an exogenous aerobic PCB-biodegrading bacterial co-culture and treated in slurry- and solid-phase laboratory conditions. Significant depletions of the spiked PCBs were observed in all microcosms of the three soils after 90 days of treatment; however, interesting yields of PCB dechlorination and detectable decreases of the original soil ecotoxicity were observed in the slurry-phase microcosms. RAMEB generally enhanced PCB-metabolism with effects which were dependent on the concentration at which it was applied, the physical-chemical nature of the amended soil, and the soil treatment conditions employed. RAMEB, which was slowly metabolized by soil microorganisms, enhanced the presence of PCBs and PCB-cometabolizing bacteria in the soil-water phase, suggesting that RAMEB enhances aerobic biodegradation of PCBs by increasing pollutant bioavailability in soil microcosms.

Aerobiosis↗