Unravelling the role of the group 6 soluble di-iron monooxygenase (SDIMO) SmoABCD in alkane metabolism and chlorinated alkane degradation.

Soluble di-iron monooxygenases (SDIMOs) are multi-component enzymes catalysing the oxidation of various substrates. These enzymes are characterized by high sequence and functional diversity that is still not well understood despite their key role in biotechnological processes including contaminant biodegradation. In this study, we analysed a mutant of Rhodoccocus aetherivorans BCP1 (BCP1-2.10) characterized by a transposon insertion in the gene smoA encoding the alpha subunit of the plasmid-located SDIMO SmoABCD. The mutant BCP1-2.10 showed a reduced capacity to grow on propane, lost the ability to grow on butane, pentane and n-hexane and was heavily impaired in the capacity to degrade chloroform and trichloroethane. The expression of the additional SDIMO prmABCD in BCP1-2.10 probably allowed the mutant to partially grow on propane and to degrade it, to some extent, together with the other short-chain n-alkanes. The complementation of the mutant, conducted by introducing smoABCD in the genome as a single copy under a constitutive promoter or within a plasmid under a thiostreptone-inducible promoter, allowed the recovery of the alkanotrophic phenotype as well as the capacity to degrade chlorinated n-alkanes. The heterologous expression of smoABCD allowed a non-alkanotrophic Rhodococcus strain to grow on pentane and n-hexane when the gene cluster was introduced together with the downstream genes encoding alcohol and aldehyde dehydrogenases and a GroEL chaperon. BCP1 smoA gene was shown to belong to the group 6 SDIMOs, which is a rare group of monooxygenases mostly present in Mycobacterium genus and in a few Rhodococcus strains. SmoABCD originally evolved in Mycobacterium and was then acquired by Rhodococcus through horizontal gene transfer events. This work extends the knowledge of the biotechnologically relevant SDIMOs by providing functional and evolutionary insights into a group 6 SDIMO in Rhodococcus and demonstrating its key role in the metabolism of short-chain alkanes and degradation of chlorinated n-alkanes.

Ammonium removal and recovery from municipal wastewater by ion exchange using a metakaolin K-based geopolymer.

Among the available technologies for ammonium removal from wastewater, ion exchange represents one of the most promising ones in the perspective to recover ammonium and produce a fertilizing product. However, the vast majority of previous studies on ammonium ion exchange did not evaluate the process robustness under real operational conditions nor optimized the desorption step. In this paper, tests of ammonium removal and recovery were conducted on a metakaolin K-based geopolymer, compared with a high-performing Italian natural zeolite in K-form. Real municipal and saline wastewater was treated in a continuous flow pilot plant equipped with a 60-cm adsorption bed (bed volume 203 mL, sorbent mass 145-173 g, empty bed contact time 10 min). Geopolymer granules showed higher performances in terms of selectivity towards ammonium, operating capacity (8.5 mgN g-1 dry adsorbent at an inlet concentration of 40 mgN L-1), bed volumes of wastewater treated at the selected breakpoint (149). Geopolymer resulted to be a cost-effective adsorbent for wastewater treatment capable to adsorb cations by ion exchange, allowing a fractionated desorption procedure that led to recover ammonium in a solution composed mainly by NH4NO3 (37%wt) and KNO3 (56%wt), potentially usable as fertilizer. The geopolymer robustness was assessed after repeated adsorption/regeneration cycles showing that the geopolymer mechanical and morphological properties did not deteriorate. The results make the tested geopolymer a very promising material for the optimization and scale-up of the ammonium recovery process in a circular economy perspective.

Regeneration and modelling of a phosphorous removal and recovery hybrid ion exchange resin after long term operation with municipal wastewater.

Adsorption represents one of the most promising process for phosphorous (P) removal and recovery from municipal wastewater, but questions about its long-term stability remain. The goals of this work were (i) to assess changes in morphology and adsorption performances of hybrid anion exchanger (HAIX) LayneRT after 2.5 years of operation in a 10 m3 d-1 demonstration plant fed with secondary-treated municipal wastewater, (ii) to optimize the LayneRT regeneration procedure, and (iii) to evaluate the suitability of the ion exchange model to describe P adsorption on LayneRT. LayneRT is composed of hydrated ferric nanoparticles dispersed in a strong base anion exchange resin. Batch and continuous flow adsorption/desorption tests were conducted with the resin used for 2.5 years, regenerated with two alternative solutions: NaOH, reactivating mainly the iron nanoparticles active sites, and NaOH + NaCl, also regenerating the active sites of the ion exchange media. The physicochemical characterization by Scanning Electron Microscope indicated that regeneration by NaOH significantly reduced the deterioration of the resin surface, even after 59 adsorption/desorption cycles. Lab-scale continuous flow tests showed that the resin regenerated with either solution featured P adsorption performances very close to that of the virgin resin. The isotherm tests showed that P adsorption by LayneRT was effectively simulated with the ion exchange model. This study confirms that LayneRT is a durable, resistant and promising media for P recovery from wastewater.

Characterization of clogging deposits in an irrigation pipeline and effect of post-aeration on clogging potential of tertiary-treated wastewater.

In Agadir, a water-scarce Moroccan region, municipal and industrial wastewater is tertiary-treated to be reused in golf courses. Wastewater reuse has been constrained by severe clogging of emitters, which caused technical and financial problems. This study aimed to perform an in-depth characterization of the treated wastewater (TWW) in relation to its susceptibility to cause clogging, and to assess the capacity of an aeration post-treatment to reduce the clogging potential. The post-treatment consisted of injecting different airflows (0-33 L/(h Lreactor) into the TWW. The structural, morphological and elemental composition of the clogging matter collected in the irrigation pipeline was characterized using scanning electron microscopy, scanning transmission electron microscopy, X-ray diffraction and X-ray energy dispersive spectroscopy. The 15-day aeration post-treatment at 16.5 L/(h Lreactor) presented the best cost-benefit ratio. Organic matter was totally degraded. Calcium was reduced by 9%, bicarbonates by 54%. The analysis of the deposits induced by the aeration post-treatment revealed a relevant decrease of the major constituents of the clogging deposits found in the irrigation pipeline. The results show the effectiveness of post-aeration in biodegrading residual organic matter and precipitating several salts, thus reducing the clogging potential.

Innovative Research Approaches to Cope with Water Security in Africa.

To achieve a water-secure world, water management should be approached from a multidimensional and integrative perspective, addressing the water-related issues of health, household supply, economics, the environment, and resilience to water-related and climate change hazards. Although water security has significantly improved since 2000 in Africa, there are still vast inequalities in access to water suitable in terms of quantity and quality, especially in rural areas. To achieve water-related sustainable development of African economies, a broad scope of innovative technological and management solutions is required, involving governments, research institutions, private sector parties, and civil society. This special series, composed of 8 papers, illustrates a selection of the most relevant results achieved by the 7 research projects selected and financed by the European Union under 2 dedicated Horizon 2020 calls in 2015: Water-5b-2015 "A coordination platform" and Water-5c-2015 "Development of water supply and sanitation technology, systems and tools, and/or methodologies." The innovations presented in this special series include both technological advancements and w'ater management approaches, given that the development of water-related technologies in developing countries needs to be integrated into water management strategies and economic instruments. This special series aims to help policy makers take informed decisions on how to implement innovative approaches to increase water security in African countries. Integr Environ Assess Manag 2020;16:853-855. © 2020 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Comparative Preliminary Evaluation of 2 In-stream Water Treatment Technologies for the Agricultural Reuse of Drainage Water in the Nile Delta.

In the Nile Delta, a complex network of canals collects drainage water from surface-irrigated fields but also from municipal wastewater. The goal of this work was to assess the technical, environmental, and financial feasibility of the upgrade of a drainage canal (DC) into either an in-stream constructed wetland (ICW) or a canalized facultative lagoon (CFL), in order to produce a water reusable in agriculture according to Egyptian law. The model-based design of the proposed technologies was derived from field experimental data for the ICW and laboratory data for the CFL. Both technologies, integrated by a sedimentation pond and a disinfection canal, led to the attainment of the water quality standards imposed by Egyptian Law 92/2013 for the reuse of drainage water. The life cycle assessment indicated that the upgrade of an existing DC to either an ICW or a CFL results in an extremely small environmental burden, ≤0.3% of that of a traditional activated sludge process. The cost-benefit analysis (CBA) was based on the assumptions that 1) farmers currently irrigate a nonfood crop (cotton) with the low-quality drainage water present in the DC, and 2) thanks to the upgrade to a ICW or CFL, farmers will irrigate a food crop characterized by a higher market price (rice). The CBA indicated that the DC upgrade to an ICW represents an attractive investment because it leads to a financial rate of return >10% over a wide range of cotton market prices. Conversely, the upgrade to a CFL is less attractive due to high investment costs. In conclusion, the upgrade of DCs to ICWs appears a promising option for the treatment of drainage canal water in the Nile Delta, thanks to the high pollutant removal performances, low cost, and negligible environmental burden. Integr Environ Assess Manag 2020;16:920-933. © 2020 SETAC.

Effect of oxygen mass transfer rate on the production of 2,3-butanediol from glucose and agro-industrial byproducts by Bacillus licheniformis ATCC9789.

BACKGROUND: 2,3-Butanediol (BD) is a largely used fossil-based platform chemical. The yield and productivity of bio-based BD fermentative production must be increased and cheaper substrates need to be identified, to make bio-based BD production more competitive. As BD bioproduction occurs under microaerobic conditions, a fine tuning and control of the oxygen transfer rate (OTR) is crucial to maximize BD yield and productivity. Very few studies on BD bioproduction focused on the use of non-pathogenic microorganisms and of byproducts as substrate. The goal of this work was to optimize BD bioproduction by the non-pathogenic strain Bacillus licheniformis ATCC9789 by (i) identifying the ranges of volumetric and biomass-specific OTR that maximize BD yield and productivity using standard sugar and protein sources, and (ii) performing a preliminary evaluation of the variation in process performances and cost resulting from the replacement of glucose with molasses, and beef extract/peptone with chicken meat and bone meal, a byproduct of the meat production industry. RESULTS: OTR optimization with an expensive, standard medium containing glucose, beef extract and peptone revealed that OTRs in the 7-15 mmol/L/h range lead to an optimal BD yield (0.43 ± 0.03 g/g) and productivity (0.91 ± 0.05 g/L/h). The corresponding optimal range of biomass-specific OTR was equal to 1.4-7.9 [Formula: see text], whereas the respiratory quotient ranged from 1.8 to 2.5. The switch to an agro-industrial byproduct-based medium containing chicken meat and bone meal and molasses led to a 50% decrease in both BD yield and productivity. A preliminary economic analysis indicated that the use of the byproduct-based medium can reduce by about 45% the BD production cost. CONCLUSIONS: A procedure for OTR optimization was developed and implemented, leading to the identification of a range of biomass-specific OTR and respiratory quotient to be used for the scale-up and control of BD bioproduction by Bacillus licheniformis. The switch to a byproduct-based medium led to a relevant decrease in BD production cost. Further research is needed to optimize the process of BD bioproduction from the tested byproduct-based medium.

Integrated technological and management solutions for wastewater treatment and efficient agricultural reuse in Egypt, Morocco, and Tunisia.

Mediterranean-African countries (MACs) face a major water crisis. The annual renewable water resources are close to the 500 m3 /capita threshold of absolute water scarcity, and water withdrawals exceed total renewable water resources by 30%. Such a low water availability curbs economic development in agriculture, which accounts for 86% of freshwater consumption. The analysis of the current situation of wastewater treatment, irrigation, and water management in MACs and of the research projects targeted to these countries indicates the need for 1) an enhanced capacity to analyze water stress, 2) the development of water management strategies capable of including wastewater reuse, and 3) development of locally adapted water treatment and irrigation technologies. This analysis shaped the MADFORWATER project (www.madforwater.eu), whose goal is to develop a set of integrated technological and management solutions to enhance wastewater treatment, wastewater reuse for irrigation, and water efficiency in agriculture in Egypt, Morocco, and Tunisia. MADFORWATER develops and adapts technologies for the production of irrigation-quality water from drainage canals and municipal, agro-industrial, and industrial wastewaters and technologies for water efficiency and reuse in agriculture, initially validated at laboratory scale, to 3 hydrological basins in the selected MACs. Selected technologies will be further adapted and validated in 4 demonstration plants of integrated wastewater treatment and reuse. Integrated strategies for wastewater treatment and reuse targeted to the selected basins are developed, and guidelines for the development of integrated water management strategies in other basins of the 3 target MACs will be produced. The social and technical suitability of the developed technologies and nontechnological tools in relation to the local context is evaluated with the participation of MAC stakeholders and partners. Guidelines on economic instruments and policies for the effective implementation of the proposed water management solutions in the target MACs will be developed. Integr Environ Assess Manag 2018;14:447-462. © 2018 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals, Inc. on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Kinetics of aerobic cometabolic biodegradation of chlorinated and brominated aliphatic hydrocarbons: A review.

This review analyses kinetic studies of aerobic cometabolism (AC) of halogenated aliphatic hydrocarbons (HAHs) from 2001-2015 in order to (i) compare the different kinetic models proposed, (ii) analyse the estimated model parameters with a focus on novel HAHs and the identification of general trends, and (iii) identify further research needs. The results of this analysis show that aerobic cometabolism can degrade a wide range of HAHs, including HAHs that were not previously tested such as chlorinated propanes, highly chlorinated ethanes and brominated methanes and ethanes. The degree of chlorine mineralization was very high for the chlorinated HAHs. Bromine mineralization was not determined for studies with brominated aliphatics. The examined research period led to the identification of novel growth substrates of potentially high interest. Decreasing performance of aerobic cometabolism were found with increasing chlorination, indicating the high potential of aerobic cometabolism in the presence of medium- and low-halogenated HAHs. Further research is needed for the AC of brominated aliphatic hydrocarbons, the potential for biofilm aerobic cometabolism processes, HAH-HAH mutual inhibition and the identification of the enzymes responsible for each aerobic cometabolism process. Lastly, some indications for a possible standardization of future kinetic studies of HAH aerobic cometabolism are provided.

Growth of Rhodococcus sp. strain BCP1 on gaseous n-alkanes: new metabolic insights and transcriptional analysis of two soluble di-iron monooxygenase genes.

Rhodococcus sp. strain BCP1 was initially isolated for its ability to grow on gaseous n-alkanes, which act as inducers for the co-metabolic degradation of low-chlorinated compounds. Here, both molecular and metabolic features of BCP1 cells grown on gaseous and short-chain n-alkanes (up to n-heptane) were examined in detail. We show that propane metabolism generated terminal and sub-terminal oxidation products such as 1- and 2-propanol, whereas 1-butanol was the only terminal oxidation product detected from n-butane metabolism. Two gene clusters, prmABCD and smoABCD-coding for Soluble Di-Iron Monooxgenases (SDIMOs) involved in gaseous n-alkanes oxidation-were detected in the BCP1 genome. By means of Reverse Transcriptase-quantitative PCR (RT-qPCR) analysis, a set of substrates inducing the expression of the sdimo genes in BCP1 were assessed as well as their transcriptional repression in the presence of sugars, organic acids, or during the cell growth on rich medium (Luria-Bertani broth). The transcriptional start sites of both the sdimo gene clusters were identified by means of primer extension experiments. Finally, proteomic studies revealed changes in the protein pattern induced by growth on gaseous- (n-butane) and/or liquid (n-hexane) short-chain n-alkanes as compared to growth on succinate. Among the differently expressed protein spots, two chaperonins and an isocytrate lyase were identified along with oxidoreductases involved in oxidation reactions downstream of the initial monooxygenase reaction step.

In situ aerobic cometabolism of chlorinated solvents: a review.

The possible approaches for in situ aerobic cometabolism of aquifers and vadose zones contaminated by chlorinated solvents are critically evaluated. Bioaugmentation of resting-cells previously grown in a fermenter and in-well addition of oxygen and growth substrate appear to be the most promising approaches for aquifer bioremediation. Other solutions involving the sparging of air lead to satisfactory pollutant removals, but must be integrated by the extraction and subsequent treatment of vapors to avoid the dispersion of volatile chlorinated solvents in the atmosphere. Cometabolic bioventing is the only possible approach for the aerobic cometabolic bioremediation of the vadose zone. The examined studies indicate that in situ aerobic cometabolism leads to the biodegradation of a wide range of chlorinated solvents within remediation times that vary between 1 and 17 months. Numerous studies include a simulation of the experimental field data. The modeling of the process attained a high reliability, and represents a crucial tool for the elaboration of field data obtained in pilot tests and for the design of the full-scale systems. Further research is needed to attain higher concentrations of chlorinated solvent degrading microbes and more reliable cost estimates. Lastly, a procedure for the design of full-scale in situ aerobic cometabolic bioremediation processes is proposed.

Development of an attached-growth process for the on-site bioremediation of an aquifer polluted by chlorinated solvents.

A procedure for the design of an aerobic cometabolic process for the on-site degradation of chlorinated solvents in a packed bed reactor was developed using groundwater from an aquifer contaminated by trichloroethylene (TCE) and 1,1,2,2-tetrachloroethane (TeCA). The work led to the selection of butane among five tested growth substrates, and to the development and characterization from the site's indigenous biomass of a suspended-cell consortium capable to degrade TCE (first order constant: 96 L gprotein(-1) day(-1) at 30 °C and 4.3 L gprotein(-1) day(-1) at 15 °C) with a 90 % mineralization of the organic chlorine. The consortium immobilization had strong effects on the butane and TCE degradation rates. The microbial community structure was slightly changed by a temperature shift from 30 to 15 °C, but remarkably affected by biomass adhesion. Given the higher TCE normalized degradation rate (0.59 day(-1) at 15 °C) and attached biomass concentration (0.13 gprotein Lbioreactor(-1) at 15 °C) attained, the porous ceramic carrier Biomax was selected as the best option for the packed bed reactor process. The low TeCA degradation rate exhibited by the developed consortium suggested the inclusion of a chemical pre-treatment based on the TeCA to TCE conversion via ß-elimination, a very fast reaction at alkaline pH. To the best of the authors' knowledge, this represents the first attempt to develop a procedure for the development of a packed bed reactor process for the aerobic cometabolism of chlorinated solvents.

A kinetic study of biohydrogen production from glucose, molasses and cheese whey by suspended and attached cells of Thermotoga neapolitana.

Batch tests of H2 production from glucose, molasses and cheese whey by suspended and immobilized cells of Thermotoga neapolitana were conducted to develop a kinetic model of the process. H2 production was inhibited by neither H2 (up to 0.7 mg L(-1)) nor O2 (up to 0.2 mg L(-1)). The H2 specific rates obtained at different substrate concentrations were successfully interpolated with Andrew's inhibition model. With glucose and molasses, biofilms performed better than suspended cells. The suspended-cell process was successfully scaled-up to a 19-L bioreactor. Assays co-fed with molasses and cheese whey led to higher H2 productivities and H2/substrate yields than the single-substrate tests. The simulation of the suspended-cell continuous-flow process indicated the potential attainment of H2 productivities higher than those of the batch tests (up to 3.6 mmol H2 h(-1) L(-1) for molasses and 0.67 mmol H2 h(-1) L(-1) for cheese whey) and allowed the identification of the optimal dilution rate.

Trichloroethylene aerobic cometabolism by suspended and immobilized butane-growing microbial consortia: a kinetic study.

A kinetic study of butane uptake and trichloroethylene (TCE) aerobic cometabolism was conducted by two suspended-cell (15 and 30°C) and two attached-cell (15 and 30°C) consortia obtained from the indigenous biomass of a TCE-contaminated aquifer. The shift from suspended to attached cells resulted in an increase of butane (15 and 30°C) and TCE (15°C) biodegradation rates, and a significant decrease of butane inhibition on TCE biodegradation. The TCE 15°C maximum specific biodegradation rate was equal to 0.011 mg(TCE ) mg(protein)(-1) d(-1) with suspended cells and 0.021 mg(TCE) mg(protein)(-1) d(-1) with attached cells. The type of mutual butane/TCE inhibition depended on temperature and biomass conditions. On the basis of a continuous-flow simulation, a packed-bed PFR inoculated with the 15 or 30°C attached-cell consortium could attain a 99.96% conversion of the studied site's average TCE concentration with a 0.4-0.5-day hydraulic residence time, with a low effect of temperature on the TCE degradation performances.

Aerobic/anaerobic/aerobic sequenced biodegradation of a mixture of chlorinated ethenes, ethanes and methanes in batch bioreactors.

A novel aerobic/anaerobic/aerobic treatment was implemented in batch reactors containing aquifer materials from a site contaminated by tetrachloroethylene (PCE), trichloroethylene (TCE), vinyl chloride (VC), 1,1,2-trichloroethane (1,1,2-TCA) and chloroform (CF). Consortia grown aerobically on methane, propane, n-pentane and n-hexane completely biodegraded the chlorinated solvent mixture, via aerobic cometabolism of VC, CF, TCE and 1,1,2-TCA, followed by PCE reductive dechlorination (RD) to 1,2-cis-dichlorothylene (cis-DCE) or TCE, and cis-DCE/TCE cometabolism in a further aerobic phase. n-Hexane was the best substrate. No electron donor was supplied for RD, which likely utilized cellular material produced during the aerobic phase. Chloride release was stoichiometric with chlorinated solvent biodegradation. According to the Lepidium sativum ecotoxicity test, a decreased toxicity was observed with propane, n-pentane and n-hexane, but not methane. A kinetic study of PCE RD allowed to estimate the PCE maximum specific rate (0.57 ± 0.07 mg mg(protein)(-1) day(-1)) and half-saturation constant (6.7 ± 1.5 mg L(-1)).

A continuous-flow approach for the development of an anaerobic consortium capable of an effective biomethanization of a mechanically sorted organic fraction of municipal solid waste as the sole substrate.

An effective mesophilic continuous anaerobic digestion process fed only with a mechanically sorted organic fraction of municipal solid waste (MS-OFMSW) was developed. During a preliminary 3-month experimental phase, the microbial consortium was acclimated toward MS-OFMSW by initially filling the reactor with cattle manure and then continuously feeding it with MS-OFMSW. The Hydraulic Retention Time (HRT) and Organic Loading Rate (OLR) were 23 days and 2.5 g/L/day, respectively. After 4 weeks, the reactor reached stationary performances (84% COD removal yield, 0.15 L(CH4)/g(COD removed) methane production yield). The acclimated consortium was then employed in a second run in which the reactor was operated under steady state conditions at the previous HRT and OLR for 73 days. The COD removal and the methane production yield increased up to 87% and 0.25 L(CH4) /g(CODremoved), respectively. The capability of the acclimated consortium to biomethanize MS-OFMSW was further studied via batch digestion experiments, carried out by inoculating the target waste with reactor effluents collected at the beginning of first run and at the end of the first and second run. The best normalized methane production (0.39 L(CH4) /g(initial COD)) was obtained with the inoculum collected at the end of the second run. Molecular analysis of the microbial community occurring in the reactor during the two sequential runs indicated that the progressive improvement of the process performances was closely related to the selection and enrichment of specific hydrolytic and acidogenic bacteria in the reactor.

Chloroform degradation by butane-grown cells of Rhodococcus aetherovorans BCP1.

The ability of a Rhodococcus aetherovorans strain, BCP1, to grow on butane and to degrade chloroform in the 0-633 microM range (0-75.5 mg l(-1)) via aerobic cometabolism was investigated by means of resting-cell assays. BCP1 degraded chloroform with a complete mineralization of the organic Cl. The resulting butane and chloroform maximum specific degradation rates were equal to 118 and 22 micromol mg(protein)(-1)day(-1), respectively. Butane inhibition on chloroform degradation was satisfactorily interpreted by means of a model of competitive inhibition, with an inhibition constant equal to 38 % of the estimated butane half-saturation constant, whereas chloroform (at 11 microM) did not inhibit butane utilization. Acetylene (1,720 microM) induced an almost complete inactivation of the degradation of both butane and chloroform, indicating that the studied cometabolic process is mediated by a monooxygenase enzyme. BCP1 proved capable of degrading vinyl chloride and 1,1,2-trichloroethane, but not 1,2-trans-dichloroethylene. BCP1 could grow on the intermediates of the most common butane metabolic pathways and on the aliphatic hydrocarbons from ethane to n-heptane. After growth on n-hexane, it was able to deplete chloroform (13 microM) with a degradation rate higher than that obtained, at the same chloroform concentration, after growth on butane.

Long-term aerobic cometabolism of a chlorinated solvent mixture by vinyl chloride-, methane- and propane-utilizing biomasses.

The aerobic cometabolic biodegradation of a mixture of chlorinated aliphatic hydrocarbons (CAHs) including vinyl chloride (VC), cis- and trans-1,2-dichloroethylene (cis-DCE, trans-DCE), trichloroethylene (TCE), 1,1,2-trichloroethane (1,1,2-TCA) and 1,1,2,2-tetrachloroethane (1,1,2,2-TeCA) was investigated at both 25 and 17 degrees C by means of bioaugmented and non-bioaugmented sediment-groundwater slurry microcosm tests. The goals of the study were (i) to study the long-term aerobic biodegradation of a CAH mixture including a high-chlorinated solvent (1,1,2,2-TeCA) generally considered non-biodegradable in aerobic conditions; (ii) to investigate the efficacy of bioaugmentation with two types of internal inocula obtained from the indigenous biomass of the studied site; (iii) to identify the CAH-degrading bacteria. VC, methane and propane were utilized as growth substrates. The non-bioaugmented microcosms were characterized, at 25 degrees C, by an average 18-day lag-time for the direct metabolism of VC (accompanied by the cometabolism of cis- and trans-DCE) and by long lag-times (36-264 days) for the onset of methane or propane utilization (associated with the cometabolism of the remaining CAHs). In the inoculated microcosms the lag-phases for the onset of growth substrate utilization and CAH cometabolism were significantly shorter (0-15 days at 25 degrees C). Biodegradation of the 6-CAH mixture was successfully continued for up to 410 days. The low-chlorinated solvents were characterized by higher depletion rates. The composition of the microbial consortium of a propane-utilizing microcosm was determined by 16s rDNA sequencing and phylotype analysis. To the best of our knowledge, this is the first study that documents the long-term aerobic biodegradation of 1,1,2,2-TeCA.

Aerobic cometabolism of chloroform by butane-grown microorganisms: long-term monitoring of depletion rates and isolation of a high-performing strain.

The focus of this microcosm study was to monitor the performances of 17 butane-utilizing microcosms during a long-term (100-250 days) aerobic cometabolic depletion of chloroform (CF). The depletion of the contaminant began after a lag-time variable between 0 and 23 days. All microcosms quickly reached a pseudo steady-state condition, in terms of biomass concentration (with an average of 9.3 x 106 CFU ml(-1)), chloroform depletion rate (5 micromol l(-1) d(-1)) and butane utilization rate (730 micromol l(-1) d(-1)). After about 100 days of CF depletion, a sudden 5- to 7-fold increase of the chloroform rate was observed in two microcosms, where the highest amount of contaminant had been depleted. In one of these high-performing microcosms, an experiment of chloroform depletion in the absence of butane resulted in the depletion of a surprisingly high amount of contaminant (765 micromolCF kg(-1) dry soil in 2 months) and in a marked selection of a single bacterial strain. Bioaugmentation assays conducted with the biomass selected in this microcosm and with a pure culture of the selected strain immediately resulted in very high chloroform depletion rates. Preliminary results of a study conducted with resting cells of the selected strain indicated that it can degrade chloroform concentrations up to 119 microM (14.2 mg l(-1)) without any sign of substrate toxicity, and that it is able to transform vinyl chloride and 1,1,2-trichloroethane.