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Renewable methanol can be used as a feedstock to generate value-added multicarbon components through fermentation technologies. Recently, researchers reported the production of isobutyric acid using methanol as an electron donor with open culture systems dominated by Eubacterium sp. and Clostridium sp. Here we report the ability of Clostridium luticellarii wild-type strain to produce isobutyric acid from methanol and CO2 & H2. When growing on methanol, the supplementation of acetic and butyric acid enhanced isobutyric acid final concentration, selectivity and production rate. A maximum of 5.04 ± 0.08 g L−1 isobutyric acid was produced at a rate of 0.420 ± 0.012 g L−1 d−1and selectivity of 0.70 electron per electron of total products in batch with acetic and butyric acid as electron acceptors. The pH was also found to be a major factor influencing isobutyric acid formation with maximal production at pH 6.50. Finally, in addition to its ability to produce isomers, C. luticellarii was able to perform C2-unit elongation from methanol. Overall, this study positions C. luticellarii as a promising platform for the production of isocarboxylic acids. [ABSTRACT FROM AUTHOR]

Summary: The ‘microbiome’ has become a buzzword. Multiple new technologies allow to gather information about microbial communities as they evolve under stable and variable environmental conditions. The challenge of the next decade will be to develop strategies to compose and manage microbiomes. Here, key aspects are considered that will be of crucial importance for future microbial technological developments. First, the need to deal not only with genotypes but also particularly with phenotypes is addressed. Microbial technologies are often highly dependent on specific core organisms to obtain the desired process outcome. Hence, it is essential to combine omics data with phenotypic information to invoke and control specific phenotypes in the microbiome. Second, the development and application of synthetic microbiomes is evaluated. The central importance of the core species is a no‐brainer, but the implementation of proper satellite species is an important route to explore. Overall, for the next decade, microbiome research should no longer almost exclusively focus on its capacity to degrade and dissipate but rather on its remarkable capability to capture disordered components and upgrade them into high‐value microbial products. These products can become valuable commodities in the cyclic economy, as reflected in the case of ‘reversed sanitation’, which is introduced here. [ABSTRACT FROM AUTHOR]

Wastewater is typically treated by the conventional activated sludge process, which suffers from an inefficient overall energy balance. The high-rate contact stabilization (HiCS) has been proposed as a promising primary treatment technology with which to maximize redirection of organics to sludge for subsequent energy recovery. It utilizes a feast-famine cycle to select for bioflocculation, intracellular storage, or both. We optimized the HiCS process for organics recovery and characterized different biological pathways of organics removal and recovery. A total of eight HiCS reactors were operated at 15°C at short solids retention times (SRT; 0.24-2.8 days), hydraulic contact times (tc; 8 and 15 min), and stabilization times (ts; 15 and 40 min). At an optimal SRT between 0.5 and 1.3 days and tc of 15 min and ts of 40 min, the HiCS system oxidized only 10% of influent chemical oxygen demand (COD) and recovered up to 55% of incoming organic matter into sludge. Storage played a minor role in the overall COD removal, which was likely dominated by aerobic biomass growth, bioflocculation onto extracellular polymeric substances, and settling. The HiCS process recovers enough organics to potentially produce 28 kWh of electricity per population equivalent per year by anaerobic digestion and electricity generation. This inspires new possibilities for energy-neutral wastewater treatment. [ABSTRACT FROM AUTHOR]

Microbial biotechnology has a long history of producing feeds and foods. The key feature of today's market economy is that protein production by conventional agriculture based food supply chains is becoming a major issue in terms of global environmental pollution such as diffuse nutrient and greenhouse gas emissions, land use and water footprint. Time has come to re-assess the current potentials of producing protein-rich feed or food additives in the form of algae, yeasts, fungi and plain bacterial cellular biomass, producible with a lower environmental footprint compared with other plant or animal-based alternatives. A major driver is the need to no longer disintegrate but rather upgrade a variety of low-value organic and inorganic side streams in our current non-cyclic economy. In this context, microbial bioconversions of such valuable matters to nutritive microbial cells and cell components are a powerful asset. The worldwide market of animal protein is of the order of several hundred million tons per year, that of plant protein several billion tons of protein per year; hence, the expansion of the production of microbial protein does not pose disruptive challenges towards the process of the latter. Besides protein as nutritive compounds, also other cellular components such as lipids (single cell oil), polyhydroxybuthyrate, exopolymeric saccharides, carotenoids, ectorines, (pro)vitamins and essential amino acids can be of value for the growing domain of novel nutrition. In order for microbial protein as feed or food to become a major and sustainable alternative, addressing the challenges of creating awareness and achieving public and broader regulatory acceptance are real and need to be addressed with care and expedience. [ABSTRACT FROM AUTHOR]

The predation of Bdellovibrio bacteriovorus on different periodontal pathogens has already been described. However, it is necessary to consider the polymicrobial nature of periodontal disease. The current study explores the predation of Bdellovibrio on oral pathogens organized in multispecies communities. The effect of the predator was evaluated on in vitro six species communities with microbial culturing. Additionally, the effect on ex vivo subgingival plaque and saliva samples from periodontitis patients was assessed. In the latter experiment results were examined with microbial culturing, quantitative polymerase chain reaction (qPCR) and denaturing gradient gel electrophoresis (DGGE). The latter technique was used to get an overview of the whole mixed microbial population. Results showed that even in more complex models, B. bacteriovorus was still able to predate on Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans . However predation on Prevotella intermedia and Porphyromonas gingivalis could not be validated in multispecies models. The effect of Bdellovibrio was not restricted to the target bacteria. Changes in the overall ecology of the different models were evident. It could be concluded that the efficiency of predation decreased when complexity of the models increased. However, B. bacteriovorus was able to attack two important oral pathogens, F. nucleatum, and A. actinomycetemcomitans, even when present in ex vivo clinical samples. These effects still have to be validated in in vivo models to see the impact of Bdellovibrio on the whole bacterial ecology. [ABSTRACT FROM AUTHOR]

The conventional activated sludge process is widely used for wastewater treatment, but to progress toward energy self-sufficiency, the wastewater treatment scheme needs to radically improve energy balances. We developed a high-rate contact stabilization (HiCS) reactor system at high sludge-specific loading rates (>2 kg bCOD kg −1 TSS d −1 ) and low sludge retention times (<1.2 d) and demonstrate that it is able to recover more chemical energy from wastewater organics than high-rate conventional activated sludge (HiCAS) and the low-rate variants of HiCS and HiCAS. The best HiCS system recovered 36% of the influent chemical energy as methane, due to the combined effects of low production of CO 2 , high sludge yield, and high methane yield of the produced sludge. The HiCS system imposed a feast-famine cycle and a putative selection pressure on the sludge micro-organisms toward substrate adsorption and storage. Given further optimization, it is a promising process for energy recovery from wastewater. [ABSTRACT FROM AUTHOR]

Europe is confronted with an increasing supply risk of critical raw materials. These can be defined as materials of which the risks of supply shortage and their impacts on the economy are higher compared to most of other raw materials. Within the framework of the EU Innovation Partnership on raw materials Initiative, a list of 14 critical materials was defined, including some bulk metals, industrial minerals, the platinum group metals and rare earth elements. To tackle the supply risk challenge, innovation is required with respect to sustainable primary mining, substitution of critical metals, and urban mining. In these three categories, biometallurgy can play a crucial role. Indeed, microbe–metal interactions have been successfully applied on full scale to win materials from primary sources, but are not sufficiently explored for metal recovery or recycling. On the one hand, this article gives an overview of the microbial strategies that are currently applied on full scale for biomining; on the other hand it identifies technologies, currently developed in the laboratory, which have a perspective for large scale metal recovery and the needs and challenges on which bio-metallurgical research should focus to achieve this ambitious goal. [ABSTRACT FROM AUTHOR]

Resources in used water are at present mainly destroyed rather than reused. Recovered nutrients can serve as raw material for the sustainable production of high value bio-products. The concept of using hydrogen and oxygen, produced by green or off-peak energy by electrolysis, as well as the unique capability of autotrophic hydrogen oxidizing bacteria to upgrade nitrogen and minerals into valuable microbial biomass, is proposed. Both axenic and mixed microbial cultures can thus be of value to implement re-synthesis of recovered nutrients in biomolecules. This process can become a major line in the sustainable “water factory” of the future. [ABSTRACT FROM AUTHOR]

Nitrification is a well-studied and established process to treat ammonia in wastewater. Although thermophilic nitrification could avoid cooling costs for the treatment of warm wastewaters, applications above 40 °C remain a significant challenge. This study tested the effect of salinity on the thermotolerance of mesophilic nitrifying sludge (34 °C). In batch tests, 5 g NaCl L increased the activity of aerobic ammonia-oxidizing bacteria (AerAOB) by 20-21 % at 40 and 45 °C. For nitrite-oxidizing bacteria (NOB), the activity remained unaltered at 40 °C, yet decreased by 83 % at 45 °C. In a subsequent long-term continuous reactor test, temperature was increased from 34 to 40, 42.5, 45, 47.5 and 50 °C. The AerAOB activity showed 65 and 37 % higher immediate resilience in the salt reactor (7.5 g NaCl L) for the first two temperature transitions and lost activity from 45 °C onwards. NOB activity, in contrast to the batch tests, was 37 and 21 % more resilient in the salt reactor for the first two transitions, while no difference was observed for the third temperature transition. The control reactor lost NOB activity at 47.5 °C, while the salt reactor only lost activity at 50 °C. Overall, this study demonstrates salt amendment as a tool for a more efficient temperature transition for mesophilic sludge (34 °C) and eventually higher nitrification temperatures. [ABSTRACT FROM AUTHOR]

Highlights: [•] Nitratation in an OLAND RBC could be controlled through the immersion level. [•] 85–89% of the O2 input was directly absorbed during the air exposure of the discs. [•] Process control strategies solely based on DO level did not seem to be effective. [ABSTRACT FROM AUTHOR]

Vertical farming is on its way to becoming an addition to conventional agricultural practices, improving sustainable food production for the growing world population under increasing climate stress. While the early development of vertical farming systems mainly focused on technological advancement through design innovation, the automation of hydroponic cultivation, and advanced LED lighting systems, more recent studies focus on the resilience and circularity of vertical farming. These sustainability objectives are addressed by investigating water quality and microbial life in a hydroponic cultivation context. Plant growth-promoting rhizobacteria (PGPR) have been shown to improve plant performance and resilience to biotic and abiotic stresses. The application of PGPRs to plant-growing media increases microbial functional diversity, creating opportunities to improve the circularity and resilience of vertical farming systems by reducing our dependency on chemical fertilizers and crop protection products. Here, we give a brief historical overview of vertical farming, review its opportunities and challenges in an economic, environmental, social, and political context, and discuss advances in exploiting the rhizosphere microbiome in hydroponic cultivation systems. [ABSTRACT FROM AUTHOR]

Microorganisms, including bacteria, archaea, algae and fungi, colonize natural building stones. Bacteria are among the most relevant colonizers, as they impact substrates in multiple forms, primarily attributed to their high diversity. They alter rock properties, induce discoloration, dissolution or precipitation, which can lead to degradation over time or in some cases, protection. Numerous studies suggested a link between rock alteration and bacteria, although there are still inconclusive conclusions. Moreover, the role of archaea remains unresolved. Classical cultivation techniques capture a fraction of bacterial and archaeal diversity. Recently, culture-independent and omics-technologies provide tools to further understand their full diversity and true role. Based on field and experimental work, this comprehensive review provides an overview of biocolonization and potential changes during the 21st Century. To better understand the role of bacteria and archaea, the focus will be on their capabilities to alter natural building stones. It also includes a short overview of methods to understand the processes and dynamics of biocolonization. The conclusions of this work will not only improve our understanding of deterioration in general, but it can also make sustainable biorestoration with bacteria the preferred choice instead of chemical and physical agents. • Bacteria and archaea can deteriorate and remediate rock substrates. • They affect rocks by biofilms, inducing discoloration, dissolution, precipitation. • The role of bacteria is established, but its relative importance is under debate. • Archaea upon building stones remain largely unexplored. • The understanding of biocolonization in a changing climate is essential. [ABSTRACT FROM AUTHOR]

Bacillus cereus comprises a highly versatile group of bacteria, which are of particular interest because of their capacity to cause disease. Emetic food poisoning is caused by the toxin cereulide produced during the growth of emetic B. cereus in food, while diarrhoeal food poisoning is the result of enterotoxin production by viable vegetative B. cereus cells in the small intestine, probably in the mucus layer and/or attached to the host's intestinal epithelium. The numbers of B. cereus causing disease are highly variable, depending on diverse factors linked to the host (age, diet, physiology and immunology), bacteria (cellular form, toxin genes and expression) and food (nutritional composition and meal characteristics). Bacillus cereus group strains show impressive ecological diversity, ranging from their saprophytic life cycle in soil to symbiotic (commensal and mutualistic) lifestyles near plant roots and in guts of insects and mammals to various pathogenic ones in diverse insect and mammalian hosts. During all these different ecological lifestyles, their toxins play important roles ranging from providing competitive advantages within microbial communities to inhibition of specific pathogenic organisms for their host and accomplishment of infections by damaging their host's tissues. [ABSTRACT FROM AUTHOR]

Abstract: In this lab-scale study, domestic wastewater is subjected to a chemical biological adsorption (A-stage), followed by treatment in an innovative roof installed parallel plate microalgae biofilm reactor for nutrient immobilization (I-stage). The A-stage process was operated at a hydraulic retention time (HRT) of 1h and a solid retention time of 1day (FeSO4 as flocculant). The I-stage, which consequently received the effluent of the A-stage process, was operated at an HRT of 1day and exposed to natural light. The overall system removed on average 74% of the total chemical oxygen demand, 82% of the total suspended solids, 67% of the total nitrogen and 96% of the total phosphorous in the wastewater. The design involves a relatively low capital and operating cost which is in the order of 0.5€/m3 wastewater treated. These aspects suggest that the A/I process can be used as a decentralized domestic wastewater treatment system. [Copyright &y& Elsevier]

Two 4-chloro-2-methylphenoxyacetic acid (MCPA)-degrading enrichment cultures selected from an aquifer on low (0.1 mg liter-1) or high (25 mg liter-1) MCPA concentrations were compared in terms of metabolic activity, community composition, population growth, and single cell physiology. Different community compositions and major shifts in community structure following exposure to different MCPA concentrations were observed using both 16S rRNA gene denaturing gradient gel electrophoresis fingerprinting and pyrosequencing. The communities also differed in their MCPA-mineralizing activities. The enrichments selected on low concentrations mineralized MCPA with shorter lag phases than those selected on high concentrations. Flow cytometry measurements revealed that mineralization led to cell growth. The presence of low-nucleic acid-content bacteria (LNA bacteria) was correlated with mineralization activity in cultures selected on low herbicide concentrations. This suggests that LNA bacteria may play a role in degradation of low herbicide concentrations in aquifers impacted by agriculture. This study shows that subpopulations of herbicide-degrading bacteria that are adapted to different pesticide concentrations can coexist in the same environment and that using a low herbicide concentration enables enrichment of apparently oligotrophic subpopulations. [ABSTRACT FROM AUTHOR]

Abstract: Two types of non-axenic algal cultures, one dominated by the freshwater microalgae Scenedesmus obliquus and the other by the marine microalgae Phaeodactylum tricornutum, were cultivated in two types of simple photobioreactor systems. The production rates, expressed on dry matter (DM) basis, were in the order of 0.12 and 0.18g DML−1 d−1 for S. obliquus and P. tricornutum respectively. The biogas potential of algal biomass was assessed by performing standardized batch digestion as well as digestion in a hybrid flow-through reactor (combining a sludge blanket and a carrier bed), the latter under mesophilic and thermophilic conditions. Biomethane potential assays revealed the ultimate methane yield (B 0) of P. tricornutum biomass to be about a factor of 1.5 higher than that of S. obliquus biomass, i.e. 0.36 and 0.24LCH4 g−1 volatile solids (VS) added respectively. For S. obliquus biomass, the hybrid flow-through reactor tests operated at volumetric organic loading rate (Bv) of 2.8gVSL−1 d−1 indicated low conversion efficiencies ranging between 26–31% at a hydraulic retention time (HRT) of 2.2days for mesophilic and thermophilic conditions respectively. When digesting P. tricornutum at a Bv of 1.9gVSL−1 d−1 at either mesophilic or thermophilic conditions and at an HRT of 2.2days, an overall conversion efficiency of about 50% was obtained. This work indicated that the hydrolysis of the algae cells is limiting the anaerobic processing of intensively grown S. obliquus and P. tricornutum biomass. [Copyright &y& Elsevier]

Abstract: The quality of drinking water is ensured by hygienic barriers and filtration steps, such as ozonation and granular activated carbon (GAC) filtration. Apart from adsorption, GAC filtration involves microbial processes that remove biodegradable organic carbon from the ozonated ground or surface water and ensures biological stability of the treated water. In this study, microbial community dynamics in were monitored during the start-up and maturation of an undisturbed pilot-scale GAC filter at 4 depths (10, 45, 80 and 115 cm) over a period of 6 months. New ecological tools, based on 16S rRNA gene-DGGE, were correlated to filter performance and microbial activity and showed that the microbial gradients developing in the filter was of importance. At 10 cm from the top, receiving the freshly ozonated water with the highest concentration of nutrients, the microbial community dynamics were minimal and the species richness remained low. However, the GAC samples at 80–115 cm showed a 2–3 times higher species richness than the 10–45 cm samples. The highest biomass densities were observed at 45–80 cm, which corresponded with maximum removal of dissolved and assimilable organic carbon. Furthermore, the start-up period was clearly distinguishable using the Lorenz analysis, as after 80 days, the microbial community shifted to an apparent steady-state condition with increased evenness. This study showed that GAC biofilter performance is not necessarily correlated to biomass concentration, but rather that an elevated functionality can be the result of increased microbial community richness, evenness and dynamics. [Copyright &y& Elsevier]

Abstract: Recent developments in host–microbe research give rise to a growing demand for rapid and accurate methods to quantify bacterial adhesion to epithelial cells. Here, we describe a new flow cytometric method to determine the amount and viability of gut bacteria, adhered to a monolayer of differentiated cells. The latter is a more relevant epithelium model than the suspended eukaryotic cells currently used in flow cytometric protocols. During the development of the method, we monitored the adhesion potential of six bacterial species and an intestinal microbial community to Caco-2 cells. The combination of SYBR Green I/propidium iodide was more efficient than carboxyfluorescein diacetate to stain the bacterial cells. In addition, a better separation between the Caco-2 background signal and viable and dead bacteria was obtained. A precise amount of Triton X-100 was used to detach adhered bacteria from Caco-2 cells and cell debris. Yet, a limited decrease in viability was observed for the intestinal microbial community treated with Triton X-100. The flow cytometric lower detection limit for pure bacterial cultures was 3.0–4.0log/mL, whereas a 5.0–5.5log/mL detection limit was obtained in the presence of Caco-2 cell background. The latter was sufficient to quantify adhered bacteria. To the best of our knowledge, this is the first description of a flow cytometric protocol that quantifies adhesion of both pure and mixed gut microbial cultures to a differentiated monolayer of Caco-2 cells and that allows to distinguish between viable and dead adhered bacteria. [Copyright &y& Elsevier]

Pellet softening reactors are used in centralized and decentralized drinking water treatment plants for the removal of calcium (hardness) through chemically induced precipitation of calcite. This is accomplished in fluidized pellet reactors, where a strong base is added to the influent to increase the pH and facilitate the process of precipitation on an added seeding material. Here we describe for the first time the opportunistic bacterial colonization of the calcite pellets in a full-scale pellet softening reactor and the functional contribution of these colonizing bacteria to the overall drinking water treatment process. ATP analysis, advanced microscopy, and community fingerprinting with denaturing gradient gel electrophoretic (DGGE) analysis were used to characterize the biomass on the pellets, while assimilable organic carbon (AOC), dissolved organic carbon, and flow cytometric analysis were used to characterize the impact of the biological processes on drinking water quality. The data revealed pellet colonization at concentrations in excess of 500 ng of ATP/g of pellet and reactor biomass concentrations as high as 220 mg of ATP/m3 of reactor, comprising a wide variety of different microorganisms. These organisms removed as much as 60% of AOC from the water during treatment, thus contributing toward the biological stabilization of the drinking water. Notably, only a small fraction (about 60,000 cells/ml) of the bacteria in the reactors was released into the effluent under normal conditions, while the majority of the bacteria colonizing the pellets were captured in the calcite structures of the pellets and were removed as a reusable product. [ABSTRACT FROM AUTHOR]

Abstract: There is an interest to understand the fate and behaviour of the food-borne pathogen Bacillus cereus in the gut, a challenging environment with a high bacterial background. We evaluated the current detection methods to select an appropriate strategy for B. cereus monitoring during gastrointestinal experiments. Application of quantitative real-time PCR (qPCR) in a gastrointestinal matrix required careful selection of the qPCR reaction and elaborate optimization of the DNA extraction protocol. Primer competition and depletion problems associated with qPCR reactions targeting general 16S rRNA gene can be avoided by the selection of a target sequence that is unique for and widespread among the target bacteria, such as the toxin gene nheB in the case of pathogenic B. cereus. Enumeration of B. cereus during the ileum phase was impossible by plating due to overgrowth by intestinal bacteria, while a carefully optimized qPCR enabled specific detection and quantification of B. cereus. On the other hand, plating allowed the distinction of viable, injured and dead bacteria and the germination of spores, which was not possible with qPCR. In conclusion, both plating and qPCR were necessary to yield the maximal information regarding the viability and physiology of the B. cereus population in various gastrointestinal compartments. [ABSTRACT FROM AUTHOR]

Abstract: Drinking water quality is routinely monitored in the distribution network but not inside households at the point of consumption. Fluctuating temperatures, residence times (stagnation), pipe materials and decreasing pipe diameters can promote bacterial growth in buildings. To test the influence of stagnation in households on the bacterial cell concentrations and composition, water was sampled from 10 separate households after overnight stagnation and after flushing the taps. Cell concentrations, measured by flow cytometry, increased (2–3-fold) in all water samples after stagnation. This increase was also observed in adenosine tri-phosphate (ATP) concentrations (2–18-fold) and heterotrophic plate counts (4–580-fold). An observed increase in cell biovolume and ATP-per-cell concentrations furthermore suggests that the increase in cell concentrations was due to microbial growth. After 5 min flushing of the taps, cell concentrations and water temperature decreased to the level generally found in the drinking water network. Denaturing gradient gel electrophoresis also showed a change in the microbial composition after stagnation. This study showed that water stagnation in household pipes results in considerable microbial changes. While hygienic risk was not directly assessed, it emphasizes the need for the development of good material validation methods, recommendations and spot tests for in-house water installations. However, a simple mitigation strategy would be a short flushing of taps prior to use. [ABSTRACT FROM AUTHOR]

The article discusses the aspects of Quorum Sensing, which was discovered in the marine bacterium Vibrio fischeri and limited to series of species. It states that the same systems were discovered to be present in manyu other Gram-negative bacteria. Furthermore, these type of bacteria uses acylated homoserine lactones (AHLs) as signal molecules that are mainly produced by a homolog.

Abstract: Because of the risk of antibiotic resistance development, there is a growing awareness that antibiotics should be used more carefully in animal production. However, a decreased use of antibiotics could result in a higher frequency of pathogenic bacteria, which in its turn could lead to a higher incidence of infections. Short-chain fatty acids (SCFAs) have long been known to exhibit bacteriostatic activity. These compounds also specifically downregulate virulence factor expression and positively influence the gastrointestinal health of the host. As a consequence, there is currently considerable interest in SCFAs as biocontrol agents in animal production. Polyhydroxyalkanoates (PHAs) are polymers of β-hydroxy short-chain fatty acids. Currently, PHAs are applied as replacements for synthetic polymers. These biopolymers can be depolymerised by many different microorganisms that produce extracellular PHA depolymerases. Interestingly, different studies provided some evidence that PHAs can also be degraded upon passage through the gastrointestinal tract of animals and consequently, adding these compounds to the feed might result in biocontrol effects similar to those described for SCFAs. [Copyright &y& Elsevier]

For more than 100 years it was believed that bacteria were the only group responsible for the oxidation of ammonia. However, recently, a new strain of archaea bearing a putative ammonia monooxygenase subunit A ( amoA) gene and able to oxidize ammonia was isolated from a marine aquarium tank. Ammonia-oxidizing archaea (AOA) were subsequently discovered in many ecosystems of varied characteristics and even found as the predominant causal organisms in some environments. Here, we summarize the current knowledge on the environmental conditions related to the presence of AOA and discuss the possible site-related properties. Considering these data, we deduct the possible niches of AOA based on pH, sulfide and phosphate levels. It is proposed that the AOA might be important actors within the nitrogen cycle in low-nutrient, low-pH, and sulfide-containing environments. [ABSTRACT FROM AUTHOR]

Abstract: 1,2-Dichloroethane (1,2-DCA) is a well-known recalcitrant groundwater contaminant. New environment-friendly approaches for the removal of 1,2-DCA that does not bring about volatilization of the compound are required. In this study, different anodophilic consortia enriched in microbial fuel cells (MFCs) operated under airtight conditions were shown to effectively degrade 1,2-DCA (up to 102mg per liter reactor volume per day), while concomitantly generating a current. An anodophilic consortium previously enriched with acetate as the electron donor changed its composition at the rate of 48% per week and increased its richness (Rr) 3-fold, upon adapting to 1,2-DCA as the new electron donor. After being stable, during 1month of operation, it removed up to 95% of the 1,2-DCA amount in the medium in the first 2weeks, while converting 43±4% of electrons available from the removal to electricity. A natural consortium from a 1,2-DCA contaminated site changed its composition at the rate of 9% per week and increased its Rr 2-fold, upon adapting to the MFC anode conditions with 1,2-DCA as the electron donor. After being stable, during 1month of operation, it removed up to 85% of the 1,2-DCA amount in the medium in the first 2weeks and the coulombic efficiency was 25±4%. The operation of the MFCs under closed circuit conditions resulted in higher 1,2-DCA removal rates than the operation under open circuit conditions, indicating that bioelectrochemical activities enhanced the removal of 1,2-DCA in the MFC anode. The production of ethylene glycol, acetate and carbon dioxide indicated that the anodophilic bacteria oxidatively metabolized 1,2-DCA, probably by means of a hydrolysis-based pathway. The results show that MFCs can be potentially used as a practically convenient technology for the biological removal of 1,2-DCA. [Copyright &y& Elsevier]

A nitrifying sequential batch reactor operated under 2-day cyclic aerobic and anoxic conditions was pulse dosed with incremental sulfide concentrations during anoxic conditions. The nitrite-oxidizing bacteria were found to be more sensitive to sulfide than the ammonia oxidizers. A maximum of nitrite-N to (nitrite-N + nitrate-N) accumulation ratio of 0.75 was obtained at an initial pulse sulfide-S concentration of 45 mg/L under pH control at 7.5 ± 0.2 and fully mixing conditions. Total ammonium nitrogen was removed almost 100% at a removal rate of 0.73 ± 0.05 g/L·day, achieved during the aerobic days of the cycles. Denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH) analyses indicated the shift in the ammonia- and nitrite-oxidizing populations triggered by sulfide addition, partial nitrification, and subsequent recovery to complete nitrification. Interestingly, archaeal amoA genes were retrieved under the conditions of sulfide addition. These results indicate that the pulse sulfide application can be used as a tool to accumulate nitrite, which is of importance for the subsequent anaerobic ammonium oxidation (anammox) process in the achievement of complete nitrogen removal. [ABSTRACT FROM AUTHOR]

In many microbial bioreactors, high shear rates result in strong attachment of microbes and dense biofilms. In this study, high shear rates were applied to enrich an anodophilic microbial consortium in a microbial fuel cell (MFC). Enrichment at a shear rate of about 120 s−1 resulted in the production of a current and power output two to three times higher than those in the case of low shear rates (around 0.3 s−1). Biomass and biofilm analyses showed that the anodic biofilm from the MFC enriched under high shear rate conditions, in comparison with that under low shear rate conditions, had a doubled average thickness and the biomass density increased with a factor 5. The microbial community of the former, as analysed by DGGE, was significantly different from that of the latter. The results showed that enrichment by applying high shear rates in an MFC can result in a specific electrochemically active biofilm that is thicker and denser and attaches better, and hence has a better performance. [ABSTRACT FROM AUTHOR]

The rate of anodic electron transfer is one of the factors limiting the performance of microbial fuel cells (MFCs). It is known that phenazine-based metabolites produced by Pseudomonas species can function as electron shuttles for Pseudomonas themselves and also, in a syntrophic association, for Gram-positive bacteria. In this study, we have investigated whether phenazine-based metabolites and their producers could be used to improve the electricity generation of a MFC operated with a mixed culture. Both anodic supernatants obtained from MFCs operated with a Pseudomonas strain (P-PCA) producing phenazine-1-carboxylic acid (PCA) and those from MFCs operated with a strain (P-PCN) producing phenazine-1-carboxamide (PCN) exerted similarly positive effects on the electricity generation of a mixed culture. Replacing supernatants of MFCs operated with a mixed culture with supernatants of MFCs operated with P-PCN could double the currents generated. Purified PCA and purified PCN had similar effects. If the supernatant of an engineered strain overproducing PCN was used, the effect could be maintained over longer time courses, resulting in a 1.5-fold increase in the production of charge. Bioaugmentation of the mixed culture MFCs using slow release tubes containing P-PCN not only doubled the currents but also maintained the effect over longer periods. The results demonstrated the electron-shuttling effect of phenazine-based compounds produced by Pseudomonas species and their capacity to improve the performance of MFCs operated with mixed cultures. [ABSTRACT FROM AUTHOR]

Previous studies revealed the abundance of Pseudomonas sp. in the microbial community of a microbial fuel cell (MFC). These bacteria can transfer electrons to the electrode via self-produced phenazine-based mediators. A MFC fed with acetate where several Pseudomonas sp. were present was found to be rich in a Gram-positive bacterium, identified as Brevibacillus sp. PTH1. Remarkably, MFCs operated with only the Brevibacillus strain in their anodes had poor electricity generation. Upon replacement of the anodic aqueous part of Brevibacillus containing MFCs with the cell-free anodic supernatants of MFCs operated with Pseudomonas sp. CMR12a, a strain producing considerable amounts of phenazine-1-carboxamide (PCN) and biosurfactants, the electricity generation was improved significantly. Supernatants of Pseudomonas sp. CMR12a_Reg, a regulatory mutant lacking the ability to produce PCN, had no similar improvement effect. Purified PCN, together with rhamnolipids as biosurfactants (1 mg L−1), could clearly improve electricity generation by Brevibacillus sp. PTH1, as well as enable this bacterium to oxidize acetate with concomitant reduction of ferric iron, supplied as goethite (FeOOH). When added alone, PCN had no observable effects on Brevibacillus’ electron transfer. This work demonstrates that metabolites produced by Pseudomonas sp. enable Gram-positive bacteria to achieve extracellular electron transfer. Possibly, this bacterial interaction is a key process in the anodic electron transfer of a MFC, enabling Brevibacillus sp. PTH1 to achieve its dominance. [ABSTRACT FROM AUTHOR]

Luminescent vibrios, bacteria belonging to the species Vibrio harveyi and closely related species, are important pathogens in aquaculture that can affect almost all types of cultured animals. Due to large-scale use of antibiotics, many luminescent vibrios have acquired (multiple) resistance, which render antibiotic treatments ineffective. One of the alternative strategies that has recently been developed to control infections caused by antibiotic-resistant bacteria is the disruption of quorum sensing, bacterial cell-to-cell communication. The quorum sensing system of V. harveyi has been studied quite intensively in vitro. Recent studies have been directed towards understanding the impact of quorum sensing and quorum sensing disruption on the virulence of luminescent vibrios towards different host organisms in vivo. This mini-review aims at discussing the current knowledge of quorum sensing in luminescent vibrios in vivo. Subsequently, quorum quenching by halogenated furanones is discussed and finally, some directions for further research are presented.The ISME Journal (2008) 2, 19–26; doi:10.1038/ismej.2007.92; published online 25 October 2007 [ABSTRACT FROM AUTHOR]

The massive (mis)use of antibiotics to control infections in aquaculture has resulted in the development of resistant strains, which have rendered antibiotic treatments ineffective. Moreover, the horizontal transfer of resistance determinants to human pathogens and the presence of antibiotic residues in aquaculture products for human consumption constitute important threats to public health. Therefore, to make the aquaculture industry more sustainable, new strategies to control infections are urgently needed. [Copyright &y& Elsevier]

In most natural environments as well as in engineered environments, such as wastewater treatment plants, ammonia-oxidizing bacteria (AOB) experience fluctuating substrate concentrations. Several physiological traits, such as low maintenance energy demand and decay rate, cell-to-cell communication, cell mobility, stable enzymes and RNAs, could allow AOB to maintain themselves under unfavourable circumstances. This review examines whether AOB possess such traits and how these traits might offer advantages over competing organisms such as heterotrophic bacteria during periods of starvation. In addition, within the AOB groups, differences exist in adaptation to and competitiveness under conditions of high or low ammonia or oxygen concentrations. Because these findings are of importance with regard to the ecology and activity of AOB in natural and engineered environments, concluding remarks are directed towards future research objectives that may clarify unanswered questions, thereby contributing to the general knowledge of the ecology and activity of ammonia oxidizers. [ABSTRACT FROM AUTHOR]

In this study, 16S rRNA- and rDNA-based denaturing gradient gel electrophoresis (DGGE) were used to study the temporal and spatial evolution of the microbial communities in a compost biofilter removing H2S and in a control biofilter without H2S loading. During the first 81 days of the experiment, the H2S removal efficiencies always exceeded 93% at loading rates between 4.1 and 30 g m−3 h−1. Afterwards, the H2S removal efficiency decreased to values between 44 and 71%. RNA-based DGGE analysis showed that H2S loading to the biofilter increased the stability of the active microbial community but decreased the activity-based diversity and evenness. The most intense band in both the RNA- and DNA-based DGGE patterns of the H2S-degrading biofilter represented the sulfur oxidizing bacterium Thiobacillus thioparus. This suggested that T. thioparus constituted a major part of the bacterial community and was an important primary degrader in the H2S-degrading biofilter. The decreasing H2S removal efficiencies near the end of the experiment were not accompanied by a substantial change of the DGGE patterns. Therefore, the decreased H2S removal was probably not caused by a failing microbiology but rather by a decrease of the mass transfer of substrates after agglutination of the compost particles. [ABSTRACT FROM AUTHOR]

In this study, we describe the effects of composting on the diversity, abundance and activity of the methanotrophic community present in the compost. Composting was allowed to proceed for 10 weeks in an in-vessel reactor. Self-heating capacity (Rottegrad) indicated that compost maturity was reached after 4 weeks. After 6 weeks, a second thermophilic phase was induced by manually increasing temperature to investigate whether or not the methanotrophs shifted back to the thermophilic population. Polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) with primers specific for type I methanotrophs of 16S rDNA and 16S rRNA were used to characterize the composition of the microbial community. Cluster and diversity analyses of RNA DGGE patterns were more sensitive than those of DNA DGGE patterns, and revealed that mesophilic and thermophilic methanotrophic communities could be differentiated. Moreover, it was seen that the diversity of the community was low during the thermophilic phase and increased during the final maturation phase. Real-time PCR analysis was also performed on the DNA and RNA extracts and showed no changes in the abundance of type I methanotrophs during the composting process (109 DNA copies/g compost). However, RNA-related activity did change, with the lowest activity (107 cDNA copies/g compost) observed during the thermophilic phase, subsequently increasing to its maximum value (109 cDNA copies/g compost), and finally decreasing during the maturation phase. This study confirmed the population dynamics, as seen for general groups such as bacteria and fungi during composting, for a very specific and sensitive group of bacteria, it is the type I methanotrophs. [Copyright &y& Elsevier]

This study investigated the feasibility of a slow-release inoculation approach as a bioaugmentation strategy for the degradation of lindane (γ-hexachlorocyclohexane [γ-HCH]). Slow-release inoculation of Sphingomonas sp. γ 1-7 was established in both liquid and soil slurry microcosms using open-ended silicone tubes in which the bacteria are encapsulated in a protective nutrient-rich matrix. The capacity of the encapsulated cells to degrade lindane under aerobic conditions was evaluated in comparison with inoculation of free-living cells. Encapsulation of cells in tubes caused the removal of lindane by adsorption to the silicone tubes but also ensured prolonged biodegradation activity. Lindane degradation persisted 2.2 and 1.4 times longer for liquid and soil slurry microcosms, respectively, than that for inoculation with free cells. While inoculation of free-living cells led to a loss in lindane-degrading activity in limited time intervals, encapsulation in tubes allowed for a more stable actively degrading community. The loss in degrading activity was linked to the loss of the linA gene, encoding γ-HCH dehydrochlorinase (LinA), which is involved in the initial steps of the lindane degradation pathway. This work shows that a slow-release inoculation approach using a catabolic strain encapsulated in open-ended tubes is a promising bioaugmentation tool for contaminated sites, as it can enhance pollutant removal and can prolong the degrading activity in comparison with traditional inoculation strategies. [ABSTRACT FROM AUTHOR]

An evolutionary algorithm was used to determine the optimal combination of parameters for transconjugant formation. As a model system, a gfp tagged TOL plasmid pWW0 was chosen to examine transfer from Pseudomonas putida to Escherichia coli. A comparison of flow cytometry results with plating and microscopy showed that the majority of transconjugants were not culturable. The transconjugant ratio therefore was determined by flow cytometry. The evolutionary algorithm showed that the optimal conditions were obtained at 28°C and at the highest nutrient concentrations. This work demonstrates that evolutionary algorithms can be used to find optimal parameter interactions in environmental microbiology. [ABSTRACT FROM AUTHOR]

Up to now, the production and role of N-acyl homoserine lactones (AHLs) in activated sludge have been poorly understood. In this study, cross-feeding assays with the reporter strains Agrobacterium tumefaciens NTL4 and Chromobacterium violaceum CV026 were used to investigate AHL signal production by municipal activated sludge samples. AHL signal production was consistently detected from municipal activated sludge when different samples were incubated on nutrient media. From one municipal activated sludge sample, 10 strains producing AHL-like auto inducers were isolated by an overlay technique. 16S rDNA-based phylogenetic analysis showed that eight of the isolates belonged to Aeromonas spp. and two to Pseudomonas spp. Box-PCR indicated that six of these Aeromonas isolates were different strains and the two Pseudomonas strains were identical. The production of AHL or AHL-like compounds by these strains was confirmed by thin layer chromatography and biosensor overlays. The six different Aeromonas strains were found to produce the same set of AHLs, including N-hexanoyl-L-homoserine lactone. These results may indicate the possible presence of AHLs in municipal activated sludge. The potential roles of AHL in this eco system are briefly discussed. [ABSTRACT FROM AUTHOR]

In the past decades, large amounts of non-insecticidal hexachlorocyclohexane (HCH) isomers (α-,β-,δ- andℇ-HCH) have been dumped as side-products of the insecticideγ-HCH (lindane). This study investigates the effect of HCH isomers on methane oxidation, an important soil function performed by methanotrophic bacteria. Both activity and structure of the methanotrophic community were assessed, using methane oxidation assays and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) respectively. Methane oxidation assays with historically polluted soils revealed that on the long-term methane oxidation was inhibited by HCH pollution. PCR-DGGE and diversity analysis based on Lorenz curves showed that the type I methanotrophic community was less evenly distributed in historically HCH-polluted soils compared with less polluted reference soils. Short-term experiments with methane-enriched consortia further demonstrated that onlyγ- andδ-isomers inhibited methane oxidation. Type I methanotrophs of methane-enriched microbial consortia that receivedγ- orδ-HCH evolved towards higher species richness. Apparently, for historically HCH-polluted soils, a narrow community remained after long-term exposure while in case of short-term exposures, methane-enriched consortia were converted into less active, but richer communities when they were stressed by the presence ofγ- orδ-HCH. This work demonstrates the importance of incorporating all isomers and possible other side-products in risk assessment studies of persistent organic pollutants and the use of structural analysis of type I methanotrophic communities as evaluating tool. [ABSTRACT FROM AUTHOR]

High-rate electron transfer toward an anode in microbial fuel cells (MFCs) has thus far not been described for bacteria- producing soluble redox mediators. To study the mechanism of electron transfer, we used a MFC isolate, Pseudomonas aeruginosa strain KRP1. Bacterial electron transfer toward the MFC anode was enabled through pyocyanin and phenazine-1-carboxamide. The presence of the anode stimulated pyocyanin production. Mutant strains, deficient in the synthesis of pyocyanin and phenazine-1 -carboxamide, were unable to achieve substantial electron transfer and reached only 5% of the wild type's power output Upon pyocyanin addition, the power output was restored to 50%. Pyocyanin was not only used by P. aeruginosa to improve electron transfer but as well enhanced electron transfer by other bacterial species. The finding that one bacterium can produce electron shuttles, which can be used also by other bacteria, to enhance electron-transfer rate and growth, has not been shown before. These findings have considerable implications with respect to the power output attainable in MFCs. [ABSTRACT FROM AUTHOR]

Abstract: The prebiotic potential of native chicory inulin was assessed in the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) by monitoring microbial community from the colon compartments, its metabolic activity and community structure. Inulin addition selected for a higher short chain fatty acid production with shifts towards propionic and butyric acid. Conventional culture-based techniques and PCR-denaturing gradient gel electrophoresis analysis showed no remarkable changes in the overall microbial community from the colon compartments of the SHIME, whereas selective effects were seen for lactic acid bacteria. Quantitative PCR with bifidobacteria-specific primers revealed a significant increase with more than 1 log CFU ml−1 from the proximal to distal colon, in contrast to culture-based techniques, which only showed a minor bifidogenic effect in the ascending colon. Our results indicate that inulin purports prebiotic effects from the proximal to distal colon and that real-time PCR is a more precise technique to detect differences in bifidobacterial populations whereas conventional culturing techniques are much more variable. [Copyright &y& Elsevier]

Disease outbreaks—some of them caused by pathogenic bacteria—are considered to be one of the largest constraints to development of the aquaculture sector. So far, antibiotics and disinfectants have only had limited success in the prevention or cure of aquatic disease. Moreover, the frequent use of biocides, especially in subtherapeutic doses, is leading to the rapid development of resistance. Therefore, there is an urgent need to develop alternative ways to control infections caused by bacterial pathogens in aquaculture. Many of these pathogens are found to control virulence factor expression by a cell-to-cell communication system. Hence, disruption of bacterial quorum sensing has been proposed as a new anti-infective strategy and several techniques that could be used to disrupt quorum sensing have been investigated. These techniques comprise (1) the inhibition of signal molecule biosynthesis, (2) the application of quorum sensing antagonists (including natural occurring as well as synthetic halogenated furanones, antagonistic quorum sensing molecules and undefined exudates of higher plants and algae), (3) the chemical inactivation of quorum sensing signals by oxidised halogen antimicrobials, (4) signal molecule biodegradation by bacterial lactonases and by bacterial and eukaryotic acylases and (5) the application of quorum sensing agonists. The few reports that deal with disruption of quorum sensing of aquatic pathogens, together with the results obtained with human and plant pathogens, indicate that this new approach has potential in fighting infections in aquaculture. However, before this new strategy can be applied in aquaculture, the impact of quorum sensing disruption on the virulence of aquatic pathogens and the impact of the proposed quorum sensing disrupting techniques on the aquaculture system of interest should be studied in more depth. [Copyright &y& Elsevier]

Microbial fuel cells hold great promise as a sustainable biotechnological solution to future energy needs. Current efforts to improve the efficiency of such fuel cells are limited by the lack of knowledge about the microbial ecology of these systems. The purposes of this study were (i) to elucidate whether a bacterial community, either suspended or attached to an electrode, can evolve in a microbial fuel cell to bring about higher power output, and (ii) to identify species responsible for the electricity generation. Enrichment by repeated transfer of a bacterial consortium harvested from the anode compartment of a biofuel cell in which glucose was used increased the output from an initial level of 0.6 W m-2 of electrode surface to a maximal level of 4.31 W m-2 (664 mV, 30.9 mA) when plain graphite electrodes were used. This result was obtained with an average loading rate of I g of glucose liter-1 day-1 and corresponded to 81% efficiency for electron transfer from glucose to electricity. Cyclic voltammetry indicated that the enhanced microbial consortium had either membrane-bound or excreted redox components that were not initially detected in the community. Dominant species of the enhanced culture were identified by denaturing gradient gel electrophoresis and culturing. The community consisted mainly of facultative anaerobic bacteria, such as Alcaligenes faecalis and Enterococcus gallinarum, which are capable of hydrogen production. Pseudomonas aeruginosa and other Pseudomonas species were also isolated. For several isolates, electrochemical activity was mainly due to excreted redox mediators, and one of these mediators, pyocyanin produced by P. aeruginosa, could be characterized. Overall, the enrichment procedure, irrespective of whether only attached or suspended bacteria were examined, selected for organisms capable of mediating the electron transfer either by direct bacterial transfer or by excretion of redox components. [ABSTRACT FROM AUTHOR]

In the absence of oxygen, a protective H2 film is formed around an Fe(0) surface, inhibiting the electron flow from this surface. Our study of anoxic corrosion of Fe(0) beads revealed that, in the presence of Shewanella oneidensis MR-1, H2 removal and precipitation of Fe mineral particles on the cell surface are determining processes for corrosion. These two biologically mediated processes were governed by cell density. H2 removal by Shewanella oneidensis was detected at cell concentrations of 1.0 × 106 live cells ml−1 and higher and H2 was electron donor for denitrification of NO3–. The removal of the protective H2 layer from Fe(0) beads by Shewanella oneidensis, resulted in an increase of Fe release out of the Fe(0) beads from 153 ± 25 mg l−1 to 196 ± 7 mg l−1 after 20 h. When the cell concentration exceeded 1.0 × 108 live cells ml−1, precipitation of iron minerals on the cell surface was characteristic for the greatest percentage of MR-1 cells, whereas micrometre-scale iron precipitates not associated with culturable cell biomass significantly decreased in number. Addition of supernatant of a corrosion assay with high cell concentration induced metabolic activity in a corrosion assay with low cell concentration, resulting in increased H2 consumption and Fe release from Fe(0) beads. Homoserine lactone-like molecules were detected in the supernatant by a bio-assay, suggesting the involvement of a quorum-sensing regulatory mechanism. [ABSTRACT FROM AUTHOR]

Plasmid pC1 of Delftia acidovorans CA28, encoding the oxidative deamination of 3-chloroaniline (3-CA), was tagged with a mini-Tn5 transposon containing the gfp gene. The labelled plasmid, designated pC1gfp, was subsequently transferred to Pseudomonas putida UWC3 and the plasmid transfer from this donor to the bacterial community in activated sludge was studied. Mating experiments were performed on both Luria–Bertani agar plates and directly in liquid activated sludge. Green fluorescent colonies appearing on mineral medium containing 3-CA as sole nitrogen source were picked up and verified to be true pC1gfp-harbouring transconjugants. REP- and BOX-PCR genomic fingerprinting revealed a large diversity in the transconjugant collection, indicating the occurrence of multiple plasmid transfer events. Remarkably, Luria–Bertani agar plate matings yielded a different set of transconjugants compared to plasmid transfers directly in liquid activated sludge. From the plate matings, mainly Aeromonas sp. was isolated, whereas D. acidovorans strains were predominant in the transconjugant collection from liquid activated sludge. The majority of the isolates showed 3-CA deamination, probably to chlorocatechol, but several pC1gfp-harbouring transconjugants performed a rapid and complete 3-CA degradation. [Copyright &y& Elsevier]

During a study of ureolytic microbial calcium carbonate (CaCO[sub 3]) precipitation by bacterial isolates collected from different environmental samples, morphological differences were observed in the large CaCO[sub 3] crystal aggregates precipitated within bacterial colonies grown on agar. Based on these differences, 12 isolates were selected for further study. We hypothesized that the striking differences in crystal morphology were the result of different microbial species or, alternatively, differences in the functional attributes of the isolates selected. Sequencing of 16S rRNA genes showed that all of the isolates were phylogenetically closely related to the Bacillus sphaericus group. Urease gene diversity among the isolates was examined by using a novel application of PCR-denaturing gradient gel electrophoresis (DGGE). This approach revealed significant differences between the isolates. Moreover, for several isolates, multiple bands appeared on the DGGE gels, suggesting the apparent presence of different urease genes in these isolates. The substrate affinities (K[sub m]) and maximum hydrolysis rates (V[sub max]) of crude enzyme extracts differed considerably for the different strains. For certain isolates, the urease activity increased up to 10-fold in the presence of 30 mM calcium, and apparently this contributed to the characteristic crystal formation by these isolates. We show that strain-specific calcification occurred during ureolytic microbial carbonate precipitation. The specificity was mainly due to differences in urease expression and the response to calcium. [ABSTRACT FROM AUTHOR]

Evaluates the short-term effects of a 3-chloroaniline (CA) shock load and bioaugmentation with a 3-CA-degrading inoculant on reactor functions. Use of denaturing gradient gel electrophoresis, FISH and real-time PCR in examining the changes in the structure, abundance and activity of the nitrifying community; Sequences of the primers and probes used in the study.

The survival and activity of microbial degradative inoculants in bioreactors is critical to obtain successful biodegradation of non- or slowly degradable pollutants. Achieving this in industrial wastewater reactors is technically challenging. We evaluated a strategy to obtain complete and stable bioaugmentation of activated sludge, which is used to treat a 3-chloroaniline (3-CA) contaminated wastewater in a lab-scale semi-continuous activated sludge system. A 3-CA metabolizing bacterium, Comamonas testosteroni strain 12, was mixed with molten agar and encapsulated in 4 mm diameter open-ended silicone tubes of 3 cm long. The tubes containing the immobilized bacteria represented about 1% of the volume of the mixed liquor. The bioaugmentation activity of a reactor containing the immobilized cells was compared with a reactor with suspended 12gfp cells. From day 25-30 after inoculation, the reactor with only suspended cells failed to completely degrade 3-CA because of a decrease in metabolic activity. In the reactors with immobilized cells, however, 3-CA continued to be removed. A mass balance indicated that ca. 10% of the degradation activity was due to the immobilized cells. Slow release of the growing embedded cells from the agar into the activated sludge medium, resulting in a higher number of active 3-CA-degrading 12 cells, was responsible for ca. 90% of the degradation. Our results demonstrate that this simple immobilization procedure was effective to maintain a 3-CA-degrading population within the activated sludge community. [ABSTRACT FROM AUTHOR]

The diversity of bacterial groups of activated sludge samples that received wastewater from four different types of industry was investigated by a nested PCR–DGGE (denaturing gradient gel electrophoresis) approach. Specific 16S rRNA primers were chosen for large bacterial groups (Bacteria and α-Proteobacteria in particular), which dominate activated sludge communities, as well as for actinomycetes, ammonium oxidisers and methanotrophs (types I and II). In addition primers for the new Acidobacterium kingdom were used to observe their community structure in activated sludge. After this first PCR amplification, a second PCR with bacterial primers yielded 16S rRNA gene fragments that were subsequently separated by DGGE, thus generating ‘group-specific DGGE patterns’. The community structure and diversity of the bacterial groups from the different samples was further analysed using different techniques, such as statistical analysis and Shannon diversity index evaluation of the band patterns. By combining the seven DGGE gels, cluster analysis, multidimensional scaling and principal component analysis clearly clustered two of the four activated sludge types separately. It was shown that the combination of molecular and statistical methods can be very useful to differentiate microbial communities. [Copyright &y& Elsevier]

SummaryIt is generally assumed that increased microbial diversity corresponds to increased catabolic potential and, hence, to better removal of metabolites and pollutants. Yet, microbial diversity, more specifically richness of species in environmental samples and sites, is difficult to assess. It is proposed to interpret this diversity more in the framework of Pareto's law, i.e. 20% of the species govern 80% of the energy flux of the ecosystem. Ecological studies should attempt to delineate the main energy fluxes and that group of species playing quantitative key roles in the system. Consequently, bioaugmentation should aim at the rearrangement of the group of organisms dominantly involved in the overall energy flux, so that specific catabolic traits necessary for the clean up of pollutants are part of that active group. For soil ecosystems, the capacity of plant roots as creators of physical and chemical discontinuity should be used more strategically to bring about such rearrangements. Overall, this paper identifies a number of ecological concepts, such as the Pareto law, the Gompertz model and plant community-induced microbial competence, which may, given careful underpinning, open new perspectives for microbial ecology and biodegradation. [ABSTRACT FROM AUTHOR]

Examines the diversity of the plasmids and of the gene tdnQ, involved in the oxidative deamination of aniline, in five bacterial strains that are able to metabolize both aniline and 3-chloroaniline (3-CA). Isolation and identification of 3-CA-metabolizing bacteria; Involvement of plasmids in aniline and 3-CA degradation; Comparison of partial tdnQ sequences.