Microcosm Assessment of a DNA Probe Applied to Aerobic Degradation of cis-1,2-Dichloroethene by Polaromonas sp. Strain JS666

A molecular biological tool based on an organism-specific DNA sequence does not necessarily indicate in situ activity but serves important functions of evaluating the potential for biodegradation and mapping the distribution of an organism. Currently, DNA-based probes are accepted as evaluative tools for site assessment. However, these techniques are far from standardized, and information on precision is usually lacking. Here, we present the development and evaluation of a DNA probe for Polaromonas sp. strain JS666, a bacterium that couples growth to aerobic oxidation of cis-1,2-dichloroethene (cDCE), and is therefore a promising candidate for bioaugmentation at sites where cDCE has accumulated in aerobic zones. The DNA probe was used in conjunction with quantitative polymerase chain reaction to track the abundance of JS666 in microcosms. This series of studies has allowed explicit resolution of the accuracy and precision of the probe and its correlation with variations in microcosm performance. We determined that the method is sufficient to monitor distribution of JS666 at bioaugmented sites. We found within environmental, mixed cultures, that the DNA target does not persist long after cell death, demonstrating that positive result from the probe is a strong indicator that degradation can occur in suitable environmental conditions. Finally, in the absence of suspected predation, the probe accurately and precisely tracks growth. Collectively, the studies appear to validate the utility of the molecular probe for site assessment in a bioaugmentation context.     

Effect of Co‐culture and Nutrients Supplementation on Bioremediation of Crude Petroleum Sludge

Ex‐situ bioremediation of real‐field crude petroleum sludge was evaluated to elucidate the role of co‐culture (bioaugmentation) and external nutrients supplementation (biostimulation) under anaerobic microenvironment. Maximum removal of total petroleum hydrocarbons (TPH) was observed by integrating biostimulation with bioaugmentation (R5, 44.01%) followed by bioaugmentation alone (R4, 34.47%), co‐substrate supplemented operations [R6, 23.36%; R3, 16.5%; R2, 9.88%] and control (R1, 4.36%). Aromatics fraction showed higher degradation in all the conditions studied. Fate of six selected polycyclic aromatic hydrocarbons (PAHs) was evaluated during bioremediation. Among these, four ring PAHs compounds showed good degradation by integration of biostimulation with bioaugmentation (R5) while bioaugmentation alone (R4) documented good degradation of three ring PAHs. Lower ring PAHs compounds showed good degradation with the application of biostimulation (R6). Fluorescent in situ hybridization (FISH) detected the presence of known PAHs degrading microorganisms viz., Bacillus, Pseudomonas, Acido bacteria, Sulphur reducing bacteria Firmicutes, etc. Application of biostimulation and bioaugmentation strategies alone or in combinations documented noticeable influence on the degradation of petroleum sludge.     

Simultaneous Organic Matter and Nitrogen Removal from Piggery Slaughterhouse Wastewater Using Bioaugmented Gaslift Membrane Bioreactor

A single bioaugmentation reactor and a side-stream gaslift membrane bioreactor combined with bioaugmentation are conducted to treat real wastewater from a centralized piggery slaughterhouse in Vietnam. The bioaugmented reactor is inoculated with heterotrophic microorganisms (Bacillus sp.) isolated from piggery slaughterhouse wastewater. The results of a single bioaugmentation experiment show high removal efficiency of chemical oxygen demand (COD) (84.8%–97.5%) and total nitrogen (TN) (69.9%–87.2%) at loading rates of 1.28–3.89 and 0.14–0.37 kg m⁻³ d⁻¹, respectively. The combined system demonstrates a significantly higher TN removal efficiency (89.0%–96.1%) (p < 0.001), more stable flux (36.0–38.4 L m⁻² h⁻¹), and transmembrane pressure (0.95–1.05 bar), and better capacity of separation of solid–liquid phases compared to the single bioaugmentation. High COD and TN removal efficiency is possibly due to assimilation and simultaneous nitrification and denitrification processes. The results of this study also indicate the feasibility and propitious efficiency of the bioaugmented gaslift membrane bioreactor for piggery slaughterhouse wastewater treatment.     

Effectiveness of bacterial inoculum and mangrove plants on remediation of sediment contaminated with polycyclic aromatic hydrocarbons

The remediation of mangrove sediment contaminated with mixed polycyclic aromatic hydrocarbons (PAHs) having 3-, 4- and 5-rings by natural attenuation, bioaugmentation, phytoremediation and its combination was compared by greenhouse microcosm studies. At Days 90 and 154, the decreases of PAHs in contaminated mangrove sediment by phytoremediation, planted with one-year old Aegiceras corniculatum, and bioaugmentation, the inoculation of PAH-degrading bacterial strains isolated from mangrove sediment, either SCSH (Mycobacterium parafortuitum) or SAFY (Sphingobium yanoikuyae), were not better than that by natural attenuation (the non-vegetated and un-inoculated microcosms). The populations of SCSH and SAFY in sediment could not be maintained even with repeated inoculation, suggesting that the two isolates were not able to compete with the indigenous microbes and had little enhancement effect. Although some PAHs were accumulated in roots, root uptake only accounted for <15% of the spiked PAHs and the effect of plants on remediation were also insignificant. At the end of the 154-day experiment, the mass balance calculation revealed that the overall losses of PAHs by phytoremediation were comparable to that by bioaugmentation but were lower than that by natural attenuation, especially for the high molecular weight PAHs. Under natural attenuation, around 90% fluorene, 80% phenanthrene, 70% fluoranthene, 68% pyrene and 32% benzo[a]pyrene in contaminated sediment were removed. These results demonstrated that the mangrove sediment itself had sufficient indigenous microorganisms capable of naturally remedying PAH contamination.     

Using DNA‐Stable Isotope Probing to Identify MTBE‐ and TBA‐Degrading Microorganisms in Contaminated Groundwater

Although the anaerobic biodegradation of methyl tert‐butyl ether (MTBE) and tert‐butyl alcohol (TBA) has been documented in the laboratory and the field, knowledge of the microorganisms and mechanisms involved is still lacking. In this study, DNA‐stable isotope probing (SIP) was used to identify microorganisms involved in anaerobic fuel oxygenate biodegradation in a sulfate‐reducing MTBE and TBA plume. Microorganisms were collected in the field using Bio‐Sep® beads amended with ¹³C₅‐MTBE, ¹³C₁‐MTBE (only methoxy carbon labeled), or ¹³C₄‐TBA. ¹³C‐DNA and ¹²C‐DNA extracted from the Bio‐Sep beads were cloned and 16S rRNA gene sequences were used to identify the indigenous microorganisms involved in degrading the methoxy group of MTBE and the tert‐butyl group of MTBE and TBA. Results indicated that microorganisms were actively degrading ¹³C‐labeled MTBE and TBA in situ and the ¹³C was incorporated into their DNA. Several sequences related to known MTBE‐ and TBA‐degraders in the Burkholderiales and the Sphingomonadales orders were detected in all three ¹³C clone libraries and were likely to be primary degraders at the site. Sequences related to sulfate‐reducing bacteria and iron‐reducers, such as Geobacter and Geothrix, were only detected in the clone libraries where MTBE and TBA were fully labeled with ¹³C, suggesting that they were involved in processing carbon from the tert‐butyl group. Sequences similar to the Pseudomonas genus predominated in the clone library where only the methoxy carbon of MTBE was labeled with ¹³C. It is likely that members of this genus were secondary degraders cross‐feeding on ¹³C‐labeled metabolites such as acetate.     

Field-Scale Evaluation of Bioaugmentation Dosage for Treating Chlorinated Ethenes

A field demonstration was performed to evaluate the impacts of bioaugmentation dosage for treatment of chlorinated ethenes in a sandy-to-silty shallow aquifer. Specifically, bioaugmentation using a commercially available Dehalococcoides (DHC)-containing culture was performed in three separate groundwater recirculation loops, with one loop bioaugmented with 3.9 × 10¹¹ DHC, the second loop bioaugmented with 3.9 × 10¹² DHC, and the third loop bioaugmented with 3.9 × 10¹³ DHC. Groundwater monitoring was performed to evaluate DHC growth and migration, dechlorination rates, and aquifer geochemistry. The loop inoculated with 3.9 × 10¹² DHC showed slower dechlorination rates and DHC migration/growth compared with the other loops. This relatively poor performance was attributed to low pH conditions. Results for the loops inoculated with 3.9 × 10¹¹ and 3.9 × 10¹³ DHC showed similar timeframes for dechlorination, as evaluated at a monitoring well approximately 10 feet downgradient of the DHC injection well. Application of a recently developed one-dimensional bioaugmentation fate and transport screening model provided a reasonable prediction of the data in these two loops. Overall, these results suggest that increasing bioaugmentation dosage does not necessarily result in decreased dechlorination timeframes in the field. The ability to predict results suggests that modeling potentially can serve as an effective tool for determining bioaugmentation dosage and predicting overall remedial timeframes.     

氧化还原条件对红枫湖沉积物磷释放影响的微尺度分析

选取贵州红枫湖为研究对象,在实验室条件下模拟了自然、好氧和厌氧条件下沉积物内源磷的释放过程,联合应用微电极技术和沉积物磷形态分析对沉积物—水界面开展了微尺度观测与研究.结果表明,厌氧条件下红枫湖沉积物总磷含量显著降低,且主要是NaOH提取态磷(NaOH-P)和残渣态磷(rest-P)含量降低所致,厌氧条件下沉积物孔隙水中磷酸盐浓度明显升高,而好氧条件下沉积物孔隙水磷酸盐浓度显著降低,反映厌氧条件显著促进了红枫湖沉积物磷释放.厌氧条件下沉积物内部溶解氧浓度下降、硫还原活动增强可能是导致NaOH-P释放的主要原因.O_2浓度的降低加速了沉积物还原作用并产生大量H2S,进而与二价铁离子形成硫化亚铁沉淀,最终导致NaOH-P(Fe-P)释放到孔隙水中.好氧条件向厌氧条件的转换可通过改变沉积物内部pH值分布和微生物活动促使rest-P释放:厌氧条件下,厌氧微生物不仅可以消耗硫酸根产生H_2S,导致pH值降低,还可消耗有机质,将有机磷转变为无机磷.上述研究结果表明,沉积物—水界面氧化还原环境可影响沉积物氧渗透深度、pH值分布、微生物活动、硫循环以及有机质降解过程,进而控制沉积物磷的形态转化与释放.联合应用微电极技术和沉积物磷形态分析对湖泊沉积物—水界面开展微尺度观测研究是揭示沉积物内源磷释放机制与控制因素的有效途径.     

Treatment of Domestic Sewage in an Anaerobic–Aerobic Fixed‐bed Reactor with Recirculation of the Liquid Phase

This study reports the performance of a combined anaerobic–aerobic packed‐bed reactor that can be used to treat domestic sewage. Initially, a bench‐scale reactor was operated in three experimental phases. In the first phase, the anaerobic reactor was operated with an average organic matter removal efficiency of 77% for a hydraulic retention time (HRT) of 10 h. In the second phase, the reactor was operated with an anaerobic stage followed by an aerobic zone, resulting in a mean value of 91% efficiency. In the third and final phase, the anaerobic–aerobic reactor was operated with recirculation of the effluent of the reactor through the anaerobic zone. The system yielded mean total nitrogen removal percentages of 65 and 75% for recycle ratios (r) of 0.5 and 1.5, respectively, and the chemical oxygen demand (COD) removal efficiencies were higher than 90%. When the pilot‐scale reactor was operated with an HRT of 12 h and r values of 1.5 and 3.0, its performance was similar to that observed in the bench‐scale unit (92% COD removal for r = 3.0). However, the nitrogen removal was lower (55% N removal for r = 3.0) due to problems with the hydrodynamics in the aerobic zone. The anaerobic–aerobic fixed‐bed reactor with recirculation of the liquid phase allows for concomitant carbon and nitrogen removal without adding an exogenous source of electron donors and without requiring any additional alkalinity supplementation.     

Using qPCR Assays to Predict Rates of Cometabolism of TCE in Aerobic Groundwater

Monitored natural attenuation (MNA) is commonly used as a remedy for trichloroethene (TCE) in anaerobic groundwater; however, MNA has not been applied to TCE contamination in aerobic groundwater. Under aerobic conditions, bacteria initiate the degradation of many organic substances with oxygenase enzymes. Several of these enzymes are known to degrade TCE through a fortuitous reaction known as cometabolism. There are commercially available qPCR assays that can determine the number of gene copies of these enzymes. If the qPCR assay could be used to predict the first-order rate constant for cometabolism of TCE, the qPCR assay could be used to screen sites to determine whether MNA was a plausible remedy for TCE contamination. This study reevaluated data from water samples that were collected from 19 wells on five sites in Minnesota, New York, and Utah. Data had previously been published on the rate constant for cometabolism of TCE in the water samples as determined by a ¹⁴C-assay and the abundance of gene copies for five enzymes that cometabolize TCE as determined using a qPCR assay. The Michaelis-Menten (Haldane) kinetic parameters for cometabolism of TCE and the abundance of DNA for the five oxygenase enzymes were used to predict the rate constant for cometabolism of TCE. The predicted rate constants were evaluated and validated by comparing them to the rate constants derived from the ¹⁴C-assay. For predicted rate constants greater than 0.003 per year, the predicted rate constants agreed with the measured rate constants within a factor of three. The qPCR assay serves as a convenient screening tool to determine whether MNA is a plausible remedy for an aerobic plume of TCE.     

Bioaugmentation and Propane Biosparging for In Situ Biodegradation of 1,4‐Dioxane

Propane biosparging and bioaugmentation were applied to promote in situ biodegradation of 1,4‐dioxane at Site 24, Vandenberg Air Force Base (VAFB), CA. Laboratory microcosm and enrichment culture testing demonstrated that although native propanotrophs appeared abundant in the shallow water‐bearing unit of the aquifer (8 to 23 ft below ground surface [bgs]), they were difficult to be enriched from a deeper water‐bearing unit (82 to 90 feet bgs). Bioaugmentation with the propanotroph Rhodococcus ruber ENV425, however, supported 1,4‐dioxane biodegradation in microcosms constructed with samples from the deep aquifer. For field testing, a propane‐biosparging system consisting of a single sparging well and four performance monitoring wells was constructed in the deep aquifer. 1,4‐dioxane biodegradation began immediately after bioaugmentation with R. ruber ENV425 (36 L; 4 × 10⁹ cells/mL), and apparent first‐order decay rates for 1,4‐dioxane ranged from 0.021 day^?1 to 0.036 day^?1. First‐order propane consumption rates increased from 0.01 to 0.05 min^?1 during treatment. 1,4‐dioxane concentrations in the sparging well and two of the performance monitoring wells were reduced from as high as 1090 µg/L to <2 µg/L, while 1,4‐dioxane concentration was reduced from 135 µg/L to 7.3 µg/L in a more distal third monitoring well. No 1,4‐dioxane degradation was observed in the intermediate aquifer control well even though propane and oxygen were present. The demonstration showed that propane biosparging and bioaugmentation can be used for in situ treatment of 1,4‐dioxane to regulatory levels.     

湖泊生态系统产甲烷与甲烷氧化微生物研究进展

湖泊生态系统是重要的大气甲烷来源,其甲烷释放量占全球自然生态系统的40%.产甲烷和甲烷氧化微生物在湖泊甲烷生产和消耗过程中发挥关键作用.本文综述了近期有关湖泊生态系统甲烷产生与氧化过程的研究进展,重点介绍产甲烷与甲烷氧化微生物在湖泊中的分布特征、代谢途径以及调控机制.现有研究表明,湖泊中甲烷的生成不仅仅依靠赋存于沉积物和水体厌氧层的产甲烷古菌,还可能来自有氧环境中其他产甲烷微生物的代谢作用.湖泊中的甲烷在脱离水体逸散至大气之前,被甲烷氧化微生物利用,转化成二氧化碳和小分子有机化合物(如甲醇、甲醛和甲酸等).除了传统依赖氧气作为电子受体的好氧氧化过程外,新近研究还揭示了多种厌氧甲烷氧化过程,包括依赖还原硫酸盐、硝酸盐和亚硝酸盐以及Fe~(3+)/Mn~(4+)等金属离子的甲烷氧化过程.文献综合分析表明,反硝化型厌氧甲烷氧化过程主要发生在淡水湖泊中,而硫酸盐还原型主要发生在高盐度或者高碱度湖泊中.水体温度、溶解氧浓度可以显著影响产甲烷与甲烷氧化微生物的丰度与群落结构,其他湖泊环境条件,如盐度、pH和有机质类型等都可能改变产甲烷与甲烷氧化微生物的分布和代谢活性.不同湖泊类型的比较研究,有助于全面掌握影响湖泊产甲烷与甲烷氧化微生物的时空分布与代谢特征的主导因素.     

Zur Schlußreinigung konzentrierter Zuckerfabrikabwässer

The most important measure is that the freshwater demand of 0.5 m³ per t beet processing is strictly kept to or fallen below, to which a catalogue of measures with ten points is presented. The wastewaters which cannot be used again have to be stored prior to the final purification in order to compensate for quantities and concentrations, aerated storage ponds being successful for small sugar factories as the purification technology. For this, three cases are described as examples: supplementary equipment of two anaerobic ponds with a rod-roll aeration, by which within two months (May to July) the BOD₃ is reduced from 1,200 to 11 mg/l. The aeration of a pond of 12.5 ha (370,000 … 500,000 m³) by means of a centrifugal aerator (45 kW) does not lead to completely aerobic conditions: Accumulation up to 350 t BOD₅, 200 t organic acids at a reduction of 100 t sulphate, 45 t of which being converted via sulphides by photoautotrophic sulphur bacteria. This combination of anaerobic and aerobic processes is very economical. At aeration with ca. 1 W/m³ in the third case in summer there are achieved wastewater loads which are of a receiving-water quality.     

Basic limnological characteristics of the shallow eutrophic lake Grimnitzsee (Brandenburg, Germany)

The medium shallow lake Grimnitzsee (maximum depth: 9.9 m; mean depth: 4.6 m; area: 7.7 · 10⁶ m²) which is situated in the biosphere reserve “Schorfheide-Chorin” in northern Brandenburg (Germany) was studied in 1994 and 1995. A bathymetric map of Grimnitzsee is given for the first time. The lake is usually polymictic although in 1994 and 1995 relatively long summer stratification was observed due to very high global radiation input. Nutrient concentration, light climate, oxygen status, phytoplankton biomass and the species composition of littoral diatoms characterize the lake as eutrophic. Special features deducible from the lake's polymictic character were the multiple development of aerobic or anaerobic strata above the sediment, the fast recovery of silicon concentration in the water column after diatom sedimentation, the importance of resuspension for the success of planktonic diatom populations, and an only moderate correlation between chlorophyll a concentration and light attenuation as well as seston dry weight probably due to the influence of suspended particles.     

Dissolved Organic Carbon in Seepage Water – Production and Transformation during Soil Passage

Dissolved organic carbon (DOC) in seepage water can combine with organic pollutants, with Al and heavy metal ions and transport them through the soil profile with a potential to contaminate groundwater. We studied the production of DOC in aerobic decomposition experiments at 8 °C and moisture close to field capacity in soils from two sites with different microbial activities (spodic dystric Cambisols with moder (SLB) and mor‐moder (SLS) layers) using ¹³C‐depleted plants of differing decomposability (Epilobium angustifolium and Calamagrostis epigeios). Additionally, we investigated the DOC transformation during soil passage in decomposition experiments and in the field for the sites SLB and SLS. For SLS, decomposition of Epilobium resulted in a cumulative CO₂ production of 14% of the added C within 128 days. Priming effects were negligible. CO₂ production for the experiments using Calamagrostis was less with 11% for SLB and 10% for SLS. Cumulative DOC production was markedly high in the Epilobium decomposition experiment, being 25 g m^–2, out of which 11 g m^–2 were Epilobium‐derived (2% of the added C). For the Calamagrostis experiments, cumulative productions of DOC and Calamagrostis‐derived DOC (0.1% of the added C for SLS and SLB) were much less. During the soil passage, much of the DOC was removed by sorption or decomposition processes. Field studies at SLS and SLB using ¹³C natural abundance showed that ¹³C distribution of soil organic matter increased with depth, probably mainly due to a discrimination of C isotopes by decomposing microorganisms. DOC, however, showed a depletion of ¹³C from –28γ PDB to –29γ (SLB at 40 cm) or –28 to –30γ (SLS at 20 cm) with depth, owing to preferential decomposition of ¹³C‐enriched substances or preferential adsorption. This study indicates that DOC production is strongly affected by litter composition and that significant changes in DOC composition may occur during its passage through a soil depth of 40 cm.     

Degradation of hydrocarbons in oxidized and reduced sediments

The oxidation-reduction status of Gulf Coast wetland sediment greatly influenced hydrocarbon breakdown. Degradation of ¹⁴C-labelled octadecane and ¹⁴C-labelled naphthalene was faster in oxidized (+500 mV) sediment compared with reduced (?200 mV) sediment. Naphthalene showed no breakdown in reduced anaerobic sediment. However, a shift from strict anaerobic to aerobic conditions markedly increased napthalene degradation. An oxidized layer of sediment overlying reduced sediment was identified in several zones in a Louisiana salt marsh. Hydrocarbon degradation is likely to be greater in the oxidized surface layer.     

The Technological Conditions for the Development of Acinetobacter for Phosphorus Removal

A simplified suspended growth integrated system with an anaerobic zone in front of the aerobic one was used for the development of the bacterium Acinetobacter for phosphorus removal. The pilot plant experimental equipment was built at the Prague municipal works. The tests were carried out on a comparative basis, the control system was operated as a completely mixed activated sludge process. In the integrated system a to phosphate removal of 80 to 86% was achieved only in a biological way. In the control system, there was no development of Acinetobacter and the biological phosphate removal was only 5%. To rise it to the extent comparative with the integrated system, high doses of ferric chloride for a simultaneous precipitation of phosphate were required. The results obtained showed that the strictly biological process in the integrated system could meet normal practice requirements, even though it is not able to compete with the simultaneous phosphate precipitation using high coagulant dosages as for as the attainable efficiency and higher values of the sludge index are concerned.     

贵州草海喀斯特高原湖泊湿地甲烷氧化菌群落特征及功能探析

草海是由喀斯特盆地积水发育而形成的一个完整的、典型的高原湖泊湿地生态系统,同时也是中国面积最大的岩溶构造湖.甲烷氧化菌作为一类能够将甲烷氧化为CO₂和水的独特微生物,其活动与生态系统中能量流动和元素循环的关键步骤密不可分.为了解贵州草海喀斯特高原湖泊湿地中甲烷氧化菌的群落结构特征及功能,利用宏基因组技术对浅水沼泽、莎草湿地、深水沉积物进行研究.结果表明,草海湿地中主要的好氧甲烷氧化菌为Methylobacter和Methylomonas,主要的厌氧甲烷氧化菌为Candidatus_Methylomirabilis_oxyfera,属于NC10门的反硝化型厌氧甲烷氧化菌.所研究的4种代谢功能基因种类多样性表现为碳代谢＞氮代谢＞硫代谢＞甲烷代谢;基于KEGG数据库共注释到6大类功能和18条与碳、甲烷、氮、硫相关的完整代谢路径.PCoA分析表明浅水沼泽和莎草湿地中甲烷氧化菌的种类和功能相似,且与深水沉积物存在差异.物种与功能相关性网络分析结果显示Methylacidiphilum_fumariolicum和Methylacidiphilum_kamchatkense与草海湖泊湿地中的各个代谢功能均具有较强的相关性.显著影响草海湿地中大多数甲烷氧化菌的群落结构的环境因素是硝酸盐、电导率、总磷和有机质.     

Abschätzung der Eignung einer molekularbiologischen Methode zur Charakterisierung der Grundwasserbiozönose

Assessing the Suitability of a Molecularbiological Method To Characterise the Microbial Populations in Groundwater A molecularbiological technique was used to characterise the bacterial community structure of groundwater habitats. This method consists of the isolation of bacterial DNA from the samples, amplification of 16S rDNA by PCR (polymerase chain reaction), and separation of the amplified DNA by DGGE (denaturing gradient gel electrophoresis). By using more specific primer combinations in the PCR instead of universal eubacterial primers, also groups of microorganisms (Proteobacteria, sulfate reducer, Archaea) were determined. The resulting DGGE patterns that reflect the microbial diversity are compared and differences or similarities evaluated. In the present studies, groundwater from different sites (bank filtrate, artificially recharged groundwater, and natural groundwater) and with changing redox milieus (aerobic, anaerobic) were investigated as well as the solid aquifer material. Besides, samples were taken from the different stages of artificial groundwater recharge, i.e., from surface water to the drain tile. Samples from groundwater derived from sites with different hydrogeochemical or hydrological conditions like bank filtrate and recharged groundwater revealed great differences in DGGE patterns indicating a characteristic species composition in these habitats, while samples taken at different times from the same groundwater showed only small seasonal variations. Clearly different patterns were also found for groundwater and the adjacent solid material as well as for anaerobic and aerobic groundwaters. Looking at artificial groundwater recharge, almost identical patterns were found in raw water and samples from gravel and sand filtration. DGGE patterns from the resulting groundwater indicated a total change in community structure during underground passage. By using group specific primers, Desulfovibrionaceae, Desulfobacteriaceae, and Archaea could be detected in anaerobic groundwaters.The molecularbiological approach described here gives an increasingly comprehensive and more precise picture of the microbial population of different environments. It is especially suitable to compare the community structure from different habitats or to analyse changes for example due to environmental stress at the same site.     

Biostimulation and Bioaugmentation to Enhance Reductive Dechlorination of TCE in a Long‐Term Flow Through Column Study

Large laboratory columns (15.2 cm diameter, 183 cm long) were fed with groundwater containing trichloroethylene (TCE), were biostimulated and bioaugmented, and were monitored for over 7.5 years. The objective of the study was to observe how the selection of the carbon and energy source, i.e., whey, Newman Zone^® standard surfactant emulsified oil and Newman Zone nonionic surfactant emulsified oil, affected the rate and extent of dechlorination. Column effluent was monitored for TCE and its degradation products, redox indicators (nitrate‐N, Fe(II), sulfate), and changes in iron mineralogy. Total bacteria and Dehalococcoides mccartyi strains were quantified using q‐PCR. Complete dechlorination was only observed in the whey treated columns, occurring 1 year after bioaugmentation with addition of a culture known to dechlorinate TCE to ethene, and 3 years later in the non‐bioaugmented column. The addition of the emulsified oils with or without bioaugmentation resulted in dechlorination only through cis‐DCE and vinyl chloride. While Dehalococcoides mccartyi strains are the only known bacteria that can fully dechlorinate TCE, their presence, either natural or augmented, was not the sole determiner of complete dechlorination. The establishment of a supporting microbial community and biogeochemistry that developed with continuous feeding of whey, in addition to the presence of D. mccartyi, were necessary to support complete reductive dechlorination. Results confirm that careful selection of a biostimulant is critical to the success of TCE dechlorination in complex soil environments.     

Upscaling microbial chemotaxis in porous media

Biodegradation is an important mechanism for contaminant reduction in groundwater environments; in fact, in situ bioremediation and bioaugmentation methods represent alternatives to traditional methods such as pump-and-treat. Microbial chemotaxis has been shown to significantly increase contaminant degradation in subsurface environments. In this work, the method of volume averaging is used to upscale the microscale chemotactic microbial transport equations in order to obtain the corresponding effective medium models for the mass balance of bacteria and the chemical attractant to which they respond. As a first approach, cellular growth/death and consumption of the attractant by chemical reaction are assumed to be negligible with respect to convective and diffusive transport mechanisms. For microorganisms, two effective coefficients are introduced, namely a total motility tensor and a total velocity vector. Our results show that, under certain conditions, these coefficients can differ considerably from the values corresponding to non-chemotactic transport. These transport coefficients show strong dependence of the microstructure of the porous medium, the fluid flow fields and the distribution of the attractant.