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The study of microbial hydrocarbons removal is of great importance for the development of future bioremediation strategies. In this study, we evaluated the removal of a gaseous mixture containing toluene, m-xylene, ethylbenzene, cyclohexane, butane, pentane, hexane and heptane in aerated stirred bioreactors inoculated with Rhodococcus erythropolis and operated under non-sterile conditions. For the real-time measurement of hydrocarbons, a novel systematic approach was implemented using Selected-Ion Flow Tube Mass Spectrometry (SIFT-MS). The effect of the carbon source (∼9.5 ppm v ) on (i) the bioreactors' performance (BR1: dosed with only cyclohexane as a single hydrocarbon versus BR2: dosed with a mixture of the 8 hydrocarbons) and (ii) the evolution of microbial communities over time were investigated. The results showed that cyclohexane reached a maximum removal efficiency (RE) of 53% ± 4% in BR1. In BR2, almost complete removal of toluene, m-xylene and ethylbenzene, being the most water-soluble and easy-to-degrade carbon sources, was observed. REs below 32% were obtained for the remaining compounds. By exposing the microbial consortium to only the five most recalcitrant hydrocarbons, REs between 45% ± 5% and 98% ± 1% were reached. In addition, we observed that airborne microorganisms populated the bioreactors and that the type of carbon source influenced the microbial communities developed. The abundance of species belonging to the genus Rhodococcus was below 10% in all bioreactors at the end of the experiments. This work provides fundamental insights to understand the complex behavior of gaseous hydrocarbon mixtures in bioreactors, along with a systematic approach for the development of SIFT-MS methods., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

The anaerobic digestion microbiome has been puzzling us since the dawn of molecular methods for mixed microbial community analysis. Monitoring of the anaerobic digestion microbiome can either take place via a non-targeted holistic evaluation of the microbial community through fingerprinting or by targeted monitoring of selected taxa. Here, we compared four different microbial community fingerprinting methods, i.e., amplicon sequencing, metaproteomics, metabolomics and cytomics, in their ability to characterise the full-scale anaerobic digestion microbiome. Cytometric fingerprinting through cytomics reflects a, for anaerobic digestion, novel, single cell-based approach of direct microbial community fingerprinting by flow cytometry. Three different digester types, i.e., sludge digesters, digesters treating agro-industrial waste and dry anaerobic digesters, each reflected different operational parameters. The α-diversity analysis yielded inconsistent results, especially for richness, across the different methods. In contrast, β-diversity analysis resulted in comparable profiles, even when translated into phyla or functions, with clear separation of the three digester types. In-depth analysis of each method's features i.e., operational taxonomic units, metaproteins, metabolites, and cytometric traits, yielded certain similar features, yet, also some clear differences between the different methods, which was related to the complexity of the anaerobic digestion process. In conclusion, cytometric fingerprinting through flow cytometry is a reliable, fast method for holistic monitoring of the anaerobic digestion microbiome, and the complementary identification of key features through other methods could give rise to a direct interpretation of anaerobic digestion process performance., (Copyright © 2021. Published by Elsevier Ltd.)

Thermophilic nitrification has been proven in lab-scale bioreactors at 50 °C. The challenge is now to develop a solution for thermophilic nitrogen removal, integrating nitrification with denitrification and aerobic carbon removal. This pioneering study aimed at a single-sludge nitrification/denitrification process at 50 °C, through exposing nitrification in a step by step approach to anoxia and/or organics. Firstly, recurrent anoxia was tolerated by a nitrifying community during long-term membrane bioreactor (MBR) operation (85 days), with high ammonium oxidation efficiencies (>98%). Secondly, five organic carbon sources did not affect thermophilic ammonium and nitrite oxidation rates in three-day aerobic batch flask incubations. Moving to long-term tests with sequencing batch reactors (SBR) and MBR (>250 days), good nitrification performance was obtained at increasing COD/N influent ratios (0, 0.5, 1, 2 and 3). Thirdly, combining nitrification, recurrent anoxia and presence of organic carbon resulted in a nitrogen removal efficiency of 92-100%, with a COD/N removed of 4.8 ± 0.6 and a nitrogen removal rate of 50 ± 14 mg N g -1 VSS d -1 . Overall, this is the first proof of principle thermophilic nitrifiers can cope with redox fluctuations (aerobic/anoxic) and the aerobic or anoxic presence of organic carbon, can functionally co-exist with heterotrophs and that single-sludge nitrification/denitrification can be achieved., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Anammox, the oxidation of ammonium with nitrite, is a key microbial process in the nitrogen cycle. Under mesophilic conditions (below 40 °C), it is widely implemented to remove nitrogen from wastewaters lacking organic carbon. Despite evidence of the presence of anammox bacteria in high-temperature environments, reports on the cultivation of thermophilic anammox bacteria are limited to a short-term experiment of 2 weeks. This study showcases the adaptation of a mesophilic inoculum to thermophilic conditions, and its characterization. First, an attached growth technology was chosen to obtain the process. In an anoxic fixed-bed biofilm bioreactor (FBBR), a slow linear temperature increase from 38 to over 48 °C (0.05-0.07 °C d -1 ) was imposed to the community over 220 days, after which the reactor was operated at 48 °C for over 200 days. Maximum total nitrogen removal rates reached up to 0.62 g N L -1 d -1 . Given this promising performance, a suspended growth system was tested. The obtained enrichment culture served as inoculum for membrane bioreactors (MBR) operated at 50 °C, reaching a maximum total nitrogen removal rate of 1.7 g N L -1 d -1 after 35 days. The biomass in the MBR had a maximum specific anammox activity of 1.1 ± 0.1 g NH 4 + -N g -1 VSS d -1 , and the growth rate was estimated at 0.075-0.19 d -1 . The thermophilic cultures displayed nitrogen stoichiometry ratios typical for mesophilic anammox: 0.93-1.42 g NO 2 - -N removed g -1 NH 4 + -N removed and 0.16-0.35 g NO 3 - -N produced g -1 NH 4 + -N removed . Amplicon and Sanger sequencing of the 16S rRNA genes revealed a disappearance of the original "Ca. Brocadia" and "Ca. Jettenia" taxa, yielding Planctomycetes members with only 94-95% similarity to "Ca. Brocadia anammoxidans" and "Ca. B. caroliniensis", accounting for 45% of the bacterial FBBR community. The long-term operation of thermophilic anammox reactors and snapshot views on the nitrogen stoichiometry, kinetics and microbial community open up the development path of thermophilic partial nitritation/anammox. A first economic assessment highlighted that treatment of sludge reject water from thermophilic anaerobic digestion of sewage sludge may become attractive., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Strain LMG 30378 T was isolated from a hydrogen-oxidizing bacteria enrichment reactor inoculated with forest soil. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that this strain belonged to the genus Achromobacter . Multilocus sequence analysis combined with sequence analysis of a 765 bp nrd A gene fragment both showed Achromobacter agilis LMG 3411 T and Achromobacter denitrificans LMG 1231 T to be the closest-related neighbours to strain LMG 30378 T . Genome sequence analysis revealed a draft genome of 6.81 Mb with a G+C content of 67.2 mol%. In silico DNA-DNA hybridization with A. denitrificans LMG 1231 T and A. agilis LMG 3411 T showed 42.7 and 42.5% similarity, respectively, confirming that strain LMG 30378 T represented a novel Achromobacter species. Phenotypic and metabolic characterization revealed acid phosphatase activity and the absence of phosphoamidase activity as distinctive features. The draft genome composes all necessary metabolic components to fix carbon dioxide and to oxidize molecular hydrogen, suggesting that strain LMG 30378 T is a key organism in the enrichment reactor. Together, these data demonstrate that strain LMG 30378 T represents a novel species of the genus Achromobacter , for which the name Achromobacter veterisilvae sp. nov. is proposed. The type strain is LMG 30378 T (=CCUG 71558 T ).

Urine is a major waste product of human metabolism and contains essential macro- and micronutrients to produce edible microorganisms and crops. Its biological conversion into a stable form can be obtained through urea hydrolysis, subsequent nitrification, and organics removal, to recover a nitrate-enriched stream, free of oxygen demand. In this study, the utilization of a microbial community for urine nitrification was optimized with the focus for space application. To assess the role of selected parameters that can impact ureolysis in urine, the activity of six ureolytic heterotrophs ( Acidovorax delafieldii, Comamonas testosteroni, Cupriavidus necator, Delftia acidovorans, Pseudomonas fluorescens, and Vibrio campbellii ) was tested at different salinities, urea, and amino acid concentrations. The interaction of the ureolytic heterotrophs with a nitrifying consortium ( Nitrosomonas europaea ATCC 19718 and Nitrobacter winogradskyi ATCC 25931) was also tested. Lastly, microgravity was simulated in a clinostat utilizing hardware for in-flight experiments with active microbial cultures. The results indicate salt inhibition of the ureolysis at 30 mS cm -1 , while amino acid nitrogen inhibits ureolysis in a strain-dependent manner. The combination of the nitrifiers with C. necator and V. campbellii resulted in a complete halt of the urea hydrolysis process, while in the case of A. delafieldii incomplete nitrification was observed, and nitrite was not oxidized further to nitrate. Nitrate production was confirmed in all the other communities; however, the other heterotrophic strains most likely induced oxygen competition in the test setup, and nitrite accumulation was observed. Samples exposed to low-shear modeled microgravity through clinorotation behaved similarly to the static controls. Overall, nitrate production from urea was successfully demonstrated with synthetic microbial communities under terrestrial and simulated space gravity conditions, corroborating the application of this process in space.

During long-term extra-terrestrial missions, food is limited and waste is generated. By recycling valuable nutrients from this waste via regenerative life support systems, food can be produced in space. Astronauts' urine can, for instance, be nitrified by micro-organisms into a liquid nitrate fertilizer for plant growth in space. Due to stringent conditions in space, microbial communities need to be be defined (gnotobiotic); therefore, synthetic rather than mixed microbial communities are preferred. For urine nitrification, synthetic communities face challenges, such as from salinity, ureolysis, and organics. In this study, a synthetic microbial community containing an AOB (Nitrosomonas europaea), NOB (Nitrobacter winogradskyi), and three ureolytic heterotrophs (Pseudomonas fluorescens, Acidovorax delafieldii, and Delftia acidovorans) was compiled and evaluated for these challenges. In reactor 1, salt adaptation of the ammonium-fed AOB and NOB co-culture was possible up to 45mScm -1 , which resembled undiluted nitrified urine, while maintaining a 44±10mgNH 4 + -NL -1 d -1 removal rate. In reactor 2, the nitrifiers and ureolytic heterotrophs were fed with urine and achieved a 15±6mg NO 3 - -NL -1 d -1 production rate for 1% and 10% synthetic and fresh real urine, respectively. Batch activity tests with this community using fresh real urine even reached 29±3mgNL -1 d -1 . Organics removal in the reactor (69±15%) should be optimized to generate a nitrate fertilizer for future space applications., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

In specific municipal and industrial cases, thermophilic wastewater treatment (>45 °C) might bring cost advantages over commonly applied mesophilic processes (10-35 °C). To develop such a novel process, one needs sound parameters on kinetics, sludge yield and sludge settleability of three heterotrophic conversions: aerobic carbon removal, denitritation and denitrification. These features were evaluated in acetate-fed sequencing batch reactors (30, 40, 50 and 60 °C). Higher temperatures were accompanied by lower sludge production and maximum specific removal rates, resulting mainly from lower maximum growth rates. Thermophilic denitritation was demonstrated for the first time, with lower sludge production (18-26%), higher nitrogen removal rates (24-92%) and lower carbon requirement (40%) compared to denitrification. Acceptable settling of thermophilic aerobic (60 °C) and anoxic biomass (50 and 60 °C) was obtained. Overall, this parameter set may catalyze the establishment of thermophilic nitrogen removal, once nitritation and nitratation are characterized. Furthermore, waters with low COD/N ratio might benefit from thermophilic nitritation/denitritation., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Anaerobic digestion of high-salinity wastewaters often results in process inhibition due to the susceptibility of the methanogenic archaea. The ability of the microbial community to deal with increased salinity levels is of high importance to ensure process perseverance or recovery after failure. The exact strategy of the microbial community to ensure process endurance is, however, often unknown. In this study, we investigated how the microbial community is able to recover process performance following a disturbance through the application of high-salinity molasses wastewater. After a stable start-up, methane production quickly decreased from 625 ± 17 to 232 ± 35 mL CH 4 L -1 d -1 with a simultaneous accumulation in volatile fatty acids up to 20.5 ± 1.4 g COD L -1 , indicating severe process disturbance. A shift in feedstock from molasses wastewater to waste activated sludge resulted in complete process recovery. However, the bacterial and archaeal communities did not return to their original composition as before the disturbance, despite similar process conditions. Microbial community diversity was recovered to similar levels as before disturbance, which indicates that the metabolic potential of the community was maintained. A mild increase in ammonia concentration after process recovery did not influence methane production, indicating a well-balanced microbial community. Hence, given the change in community composition following recovery after salinity disturbance, it can be assumed that microbial community redundancy was the major strategy to ensure the continuation of methane production, without loss of functionality or metabolic flexibility., (Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Anaerobic digestion is a well-established microbial-based technology for the treatment of organic waste streams and subsequent biogas recovery. A robust and versatile microbial community to ensure overall stability of the process is essential. Four full-scale anaerobic digestion plants were followed for one year to link operational characteristics with microbial community composition and structure. Similarities between digesters, community dynamics and co-occurrence between bacteria and archaea within each digester were analysed. Free ammonia concentration (>200 mg N L -1 ) and conductivity (>30 mS cm -1 ) hindered acetoclastic methanogenesis by Methanosaetaceae. Thus, methanogenesis was pushed to the hydrogenotrophic pathway carried out by Methanobacteriales and Methanomicrobiales. Firmicutes dominated the overall bacterial community in each of the digesters (>50%), however, principal coordinate analysis of Bray-Curtis indices showed that each of the four digesters hosted a unique microbial community. The uniqueness of this community was related to two phylotypes belonging to the Syntrophomonas genus (Phy32 and Phy34) and to one unclassified bacterium (Phy2), which could both be considered marker populations in the community. A clear differentiation in co-occurrence of methanogens with several bacteria was observed between the different digesters. Our results demonstrated that full-scale anaerobic digestion plants show constant dynamics and co-occurrence patterns in function of time, but are unique in terms of composition, related to the presence of marker populations., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The increasing production of nitrogen-containing fertilizers is crucial to meet the global food demand, yet high losses of reactive nitrogen associated with the food production/consumption chain progressively deteriorate the natural environment. Currently, mesophilic nitrogen-removing microbes eliminate nitrogen from wastewaters. Although thermophilic nitrifiers have been separately enriched from natural environments, no bioreactors are described that couple these processes for the treatment of nitrogen in hot wastewaters. Samples from composting facilities were used as inoculum for the batch-wise enrichment of thermophilic nitrifiers (350 days). Subsequently, the enrichments were transferred to a bioreactor to obtain a stable, high-rate nitrifying process (560 days). The community contained up to 17% ammonia-oxidizing archaea (AOAs) closely related to 'Candidatus Nitrososphaera gargensis', and 25% nitrite-oxidizing bacteria (NOBs) related to Nitrospira calida. Incorporation of (13)C-derived bicarbonate into the respective characteristic membrane lipids during nitrification supported their activity as autotrophs. Specific activities up to 198±10 and 894±81 mg N g(-1) VSS per day for AOAs and NOBs were measured, where NOBs were 33% more sensitive to free ammonia. The NOBs were extremely sensitive to free nitrous acid, whereas the AOAs could only be inhibited by high nitrite concentrations, independent of the free nitrous acid concentration. The observed difference in product/substrate inhibition could facilitate the development of NOB inhibition strategies to achieve more cost-effective processes such as deammonification. This study describes the enrichment of autotrophic thermophilic nitrifiers from a nutrient-rich environment and the successful operation of a thermophilic nitrifying bioreactor for the first time, facilitating opportunities for thermophilic nitrogen removal biotechnology.

Urine contains the majority of nutrients in urban wastewaters and is an ideal nutrient recovery target. In this study, stabilization of real undiluted urine through nitrification and subsequent microalgae cultivation were explored as strategy for biological nutrient recovery. A nitrifying inoculum screening revealed a commercial aquaculture inoculum to have the highest halotolerance. This inoculum was compared with municipal activated sludge for the start-up of two nitrification membrane bioreactors. Complete nitrification of undiluted urine was achieved in both systems at a conductivity of 75mScm(-1) and loading rate above 450mgNL(-1)d(-1). The halotolerant inoculum shortened the start-up time with 54%. Nitrite oxidizers showed faster salt adaptation and Nitrobacter spp. became the dominant nitrite oxidizers. Nitrified urine as growth medium for Arthrospira platensis demonstrated superior growth compared to untreated urine and resulted in a high protein content of 62%. This two-stage strategy is therefore a promising approach for biological nutrient recovery., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Anaerobic digestion is regarded as a key environmental technology in the present and future bio-based economy. The microbial community completing the anaerobic digestion process is considered complex, and several attempts already have been carried out to determine the key microbial populations. However, the key differences in the anaerobic digestion microbiomes, and the environmental/process parameters that drive these differences, remain poorly understood. In this research, we hypothesized that differences in operational parameters lead to a particular composition and organization of microbial communities in full-scale installations. A total of 38 samples were collected from 29 different full-scale anaerobic digestion installations, showing constant biogas production in function of time. Microbial community analysis was carried out by means of amplicon sequencing and real-time PCR. The bacterial community in all samples was dominated by representatives of the Firmicutes, Bacteroidetes and Proteobacteria, covering 86.1 ± 10.7% of the total bacterial community. Acetoclastic methanogenesis was dominated by Methanosaetaceae, yet, only the hydrogenotrophic Methanobacteriales correlated with biogas production, confirming their importance in high-rate anaerobic digestion systems. In-depth analysis of operational and environmental parameters and bacterial community structure indicated the presence of three potential clusters in anaerobic digestion. These clusters were determined by total ammonia concentration, free ammonia concentration and temperature, and characterized by an increased relative abundance of Bacteroidales, Clostridiales and Lactobacillales, respectively. None of the methanogenic populations, however, could be significantly attributed to any of the three clusters. Nonetheless, further experimental research will be required to validate the existence of these different clusters, and to which extent the presence of these clusters relates to stable or sub-optimal anaerobic digestion., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Kitchen waste and molasses are organic waste streams with high organic content, and therefore are interesting substrates for renewable energy production by means of anaerobic digestion. Both substrates, however, often cause inhibition of the anaerobic digestion process, when treated separately, hence, co-digestion with other substrates is required to ensure stable methane production. In this research, A-sludge (sludge harvested from a high rate activated sludge system) was used to stabilize co-digestion with kitchen waste or molasses. Lab-scale digesters were fed with A-sludge and kitchen waste or molasses for a total period of 105 days. Increased methane production values revealed a stabilizing effect of concentrated A-sludge on kitchen waste digestion. Co-digestion of molasses with A-sludge also resulted in a higher methane production. Volumetric methane production rates up to 1.53 L L(-1) d(-1) for kitchen waste and 1.01 L L(-1) d(-1) for molasses were obtained by co-digestion with A-sludge. The stabilizing effect of A-sludge was attributed to its capacity to supplement various nutrients. Microbial community results demonstrated that both reactor conditions and substrate composition determined the nature of the bacterial community, although there was no direct influence of micro-organisms in the substrate itself, while the methanogenic community profile remained constant as long as optimal conditions were maintained., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The prerequisite to the development success of the novel mainstream processes partial nitritation/anammox is the out-selection of nitrite oxidizing bacteria (NOB). A recent study suggested that this could be achieved through NO production by ammonium oxidizing bacteria under cyclic oxic-anoxic conditions. Indeed, it is known that among NOB, Nitrobacter species are reversibly inhibited by NO. However, the effect of NO on the activity of the NOB genus Nitrospira is not studied so far. Such an understanding is needed, since Nitrospira related NOB are mostly prevailing in sewage treatment plants. This study quantified the effect of NO on the nitratation activity of sludge types with different Nitrobacter/Nitrospira ratios. In an oxic bubbling column, a dosage of 4.4 mg NO L(-1) d(-1) (∼2 μg NO-N L(-1) in liquid phase) inhibited the Nitrobacter dominated sludge with 24%. For the Nitrospira dominated sludge types, the inhibition was strongly correlated with the nitrite half saturation constant (K(s)) ranging from 0% to 30-50% and 60-80% inhibition of the nitrite oxidation for K(s) of 0.72, 0.36 and 0.06 mg NO2(-)-N L(-1), respectively. This study showed that nitrifying communities with high affinity for nitrite and low specific nitrite oxidation rates (K-strategists) can be strongly inhibited by NO. The degree of inhibition could be confirmed in a set-up with NO dosage through an artificial alginate-based biofilm, ensuring a more direct contact between NO and the microorganisms., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The development of thermophilic nitrogen removal strategies will facilitate sustainable biological treatment of warm nitrogenous wastewaters. Thermophilic denitrification was extensively compared to mesophilic denitrification for the first time in this study. Two sequential batch reactors (SBR) at 34 °C and 55 °C were inoculated with mesophilic activated sludge (26 °C), fed with synthetic influent in a first phase. Subsequently, the carbon source was switched from acetate to molasses, whereas in a third phase, the nitrate source was fertilizer industry wastewater. The denitrifying sludge maintained its activity at 55 °C, resulting in an immediate process start-up, obtaining nitrogen removal rates higher than 500 mg N g(-1) VSS d(-1) in less than one week. Although the mesophilic SBR showed twice as high specific nitrogen removal rates, the maximum thermophilic denitrifying activity in this study was nearly 10 times higher than the activities reported thus far. The thermophilic SBR moreover had a 73% lower sludge volume index, a 45% lower sludge production and a higher resilience towards a change in carbon source compared with the mesophilic SBR. The higher resilience was potentially related to a higher microbial diversity and evenness of the thermophilic community at the end of the synthetic feeding period. The thermophilic microbial community showed a higher similarity over the different feeding periods implying a more stable community. Overall, this study showed the capability of mesophilic denitrifiers to maintain their activity after a large temperature increase. Existing mesophilic process systems with cooling for the treatment of warm wastewaters could thus efficiently be converted to thermophilic systems with low sludge production and good settling properties., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Microalgal bacterial flocs in sequencing batch reactors (MaB-floc SBRs) represent a novel approach to wastewater treatment. In this approach, mechanical aeration is replaced by photosynthetic aeration and MaB-floc settling separates the treated wastewater from the produced biomass. However, its technical potential for industrial wastewaters needs to be shown. Therefore, wastewaters of aquaculture, manure treatment, food-processing and chemical industry were treated in MaB-floc SBRs. This treatment resulted in significantly different nutrient removal rates and effluent qualities among wastewaters. A high MaB-floc production was obtained for all wastewaters, ranging from 0.14 to 0.26g total suspended solids Lreactor(-1)day(-1). A major advantage of MaB-flocs is the harvesting via a filter press with a large pore size of 200μm, resulting in MaB-floc recoveries of 79-99% and cakes containing 12-21% dry matter. These results may contribute to evolving MaB-floc SBRs as a valuable remediation strategy, especially for aquaculture and food-processing wastewaters., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Sequencing batch reactors with microalgal bacterial flocs (MaB-floc SBRs) are a novel approach for photosynthetic aerated wastewater treatment based on bioflocculation. To assess their technical potential for aquaculture wastewater treatment in Northwest Europe, MaB-floc SBRs were up-scaled from indoor photobioreactors of 4 L over 40 and 400 L to a 12 m(3) outdoor raceway pond. Scale-up decreased the nutrient removal efficiencies with a factor 1-3 and the volumetric biomass productivities with a factor 10-13. Effluents met current discharge norms, except for nitrite and nitrate. Flue gas sparging was needed to decrease the effluent pH. Outdoor MaB-flocs showed enhanced settling properties and an increased ash and chlorophyll a content. Bioflocculation enabled successful harvesting by gravity settling and dewatering by filtering at 150-250 μm. Optimisation of nitrogen removal and biomass valorisation are future challenges towards industrial implementation of MaB-floc SBRs for aquaculture wastewater treatment., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

With oxygen supply playing a crucial role in an oxygen-limited autotrophic nitrification/denitrification (OLAND) rotating biological contactor (RBC), its controlling factors were investigated in this study. Disc rotation speeds (1.8 and 3.6rpm) showed no influence on the process performance of a lab-scale RBC, although abiotic experiments showed a significant effect on the oxygenation capacity. Estimations of the biological oxygen uptake rate revealed that 85-89% of the oxygen was absorbed by the microorganisms during the air exposure of the discs. Indeed, increasing the disc immersion (50 to 75-80%) could significantly suppress undesired nitratation, on the short and long term. The presented results demonstrated that nitratation could be controlled by the immersion level and revealed that oxygen control in an OLAND RBC should be predominantly based on the atmospheric exposure percentage of the discs., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Anaerobic digestion is an environmental key technology in the future bio-based economy. To achieve functional stability, a minimal microbial community diversity is required. This microbial community should also have a certain 'elasticity', i.e. the ability to rapidly adapt to suboptimal conditions or stress. In this study it was evaluated whether a higher degree of functional stability could be achieved by changing the feeding pattern, which can change the evenness, dynamics and richness of the bacterial community. The first reactor (CSTR stable ) was fed on daily basis, whereas the second reactor (CSTR dynamic ) was fed every 2 days. Average biogas production was 0.30 l CH4 l(-1) day(-1) in both reactors, although daily variation was up to four times higher in the CSTR dynamic compared with the CSTR stable during the first 50 days. Bacterial analysis revealed that this CSTR dynamic had a two times higher degree of bacterial community dynamics. The CSTR dynamic also appeared to be more tolerant to an organic shock load of 8 g COD l(-1) and ammonium levels up to 8000 mg TAN l(-1). These results suggest that the regular application of a limited pulse of organic material and/or a variation in the substrate composition might promote higher functional stability in anaerobic digestion., (© 2013 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.)

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 (FeSO(4) 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€/m(3) wastewater treated. These aspects suggest that the A/I process can be used as a decentralized domestic wastewater treatment system., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The biomass of industrially grown Phaeodactylum tricornutum was subjected in a novel way to bio-methanation at 33°C, i.e., in an anaerobic membrane bioreactor (AnMBR) at a hydraulic retention time of 2.5 days, at solid retention times of 20 to 10 days and at loading rates in the range of 2.6-5.9 g biomass-COD L(-1) day(-1) with membrane fluxes ranging from 1 to 0.8 L m(-2) h(-1). The total COD recovered as biogas was in the order of 52%. The input suspension was converted to a clear effluent rich in total ammonium nitrogen (546 mg TAN L(-1)) and phosphate (141 mg PO(4)-P L(-1)) usable as liquid fertilizer. The microbial community richness, dynamics, and organization in the reactor were interpreted using the microbial resource management approach. The AnMBR communities were found to be moderate in species richness and low in dynamics and community organization relative to UASB and conventional CSTR sludges. Quantitative polymerase chain reaction analysis revealed that Methanosaeta sp. was the dominant acetoclastic methanogen species followed by Methanosarcina sp. This work demonstrated that the use of AnMBR for the digestion of algal biomass is possible. The fact that some 50% of the organic matter is not liquefied means that the algal particulates in the digestate constitute a considerable fraction which should be valorized properly, for instance as slow release organic fertilizer. Overall, 1 kg of algae dry matter (DM) could be valorized in the form of biogas ( euro 2.07), N and P in the effluent (euro 0.02) and N and P in the digestate (euro 0.04), thus totaling about euro 2.13 per kilogram algae DM.

Although microalgae are promising for a cradle-to-cradle design approach of sewage treatment, their application is hampered by high harvesting costs and low C:N ratios of sewage. Therefore, the potential of microalgal bacterial flocs (MaB-flocs) was investigated for the secondary treatment of sewage supplemented with different flue gas flow rates (FGFRs) from a coal power plant. Effluent (N, P, turbidity and pH) and off gas discharge levels (NO(x), SO(x)) met the European discharge limits with a hydraulic retention time of only 0.67 days and an FGFR of 0.6Lh(-1) (0.0025 vvm). This FGFR provided sufficient carbon and resulted in removal efficiencies of 48 ± 7% CO(2), 87 ± 5% NO(x) and 99 ± 1% SO(2). MaB-flocs settled fast reaching up to a density of 19 g VSSL(-1). High biomass productivities (0.18 gL(-1)day(-1)) were obtained under a low light intensity. This successful reactor performance indicates the large potential for the industrial application of MaB-flocs for flue gas sparged sewage treatment., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Background: Anaerobic oxidation of methane coupled to sulphate reduction (SR-AOM) prevents more than 90% of the oceanic methane emission to the atmosphere. In a previous study, we demonstrated that the high methane pressure (1, 4.5, and 8 MPa) stimulated in vitro SR-AOM activity. However, the information on the effect of high-pressure on the microbial community structure and architecture was still lacking., Results: In this study we analysed the long-term enrichment (286 days) of this microbial community, which was mediating SR-AOM in a continuous high-pressure bioreactor. 99.7% of the total biovolume represented cells in the form of small aggregates (diameter less then 15 μm). An increase of the total biovolume was observed (2.5 times). After 286 days, the ANME-2 (anaerobic methanotrophic archaea subgroup 2) and SRB (sulphate reducing bacteria) increased with a factor 12.5 and 8.4, respectively., Conclusion: This paper reports a net biomass growth of communities involved in SR-AOM, incubated at high-pressure.

Marine micro-organisms have been playing highly diverse roles over evolutionary time: they have defined the chemistry of the oceans and atmosphere. During the last decades, the bioreactors with novel designs have become an important tool to study marine microbiology and ecology in terms of: marine microorganism cultivation and deep-sea bioprocess characterization; unique bio-chemical product formation and intensification; marine waste treatment and clean energy generation. In this review we briefly summarize the current status of the bioreactor technology applied in marine microbiology and the critical parameters to take into account during the reactor design. Furthermore, when we look at the growing population, as well as, the pollution in the coastal areas of the world, it is urgent to find sustainable practices that beneficially stimulate both the economy and the natural environment. Here we outlook a few possibilities where innovative bioreactor technology can be applied to enhance energy generation and food production without harming the local marine ecosystem., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The incidence and fate of pharmaceuticals in the water cycle impose a growing concern for the future reuse of treated water. Because of the recurrent global use of drugs such as Acetaminophen (APAP), an analgesic and antipyretic drug, they are often detected in wastewater treatment plant (WWTP) effluents, receiving surface waters and drinking water resources. In this study, the removal of APAP has been demonstrated in a membrane bioreactor (MBR) fed with APAP as the sole carbon source. After 16 days of operation, at a hydraulic retention time (HRT) of 5 days, more than 99.9% removal was obtained when supplying a synthetic WWTP effluent with 100 μg APAP L(-1). Batch experiments indicated no sorption of APAP to the biomass, no influence of the WWTP effluent matrix, and the capability of the microbial consortium to remove APAP at environmentally relevant concentrations (8.3 μg APAP L(-1)). Incubation with allylthiourea, an ammonia monooxygenase inhibitor, demonstrated that the APAP removal was mainly associated with heterotrophic bacteria and not with the ammonia-oxidizing bacteria. Two APAP degrading strains were isolated from the MBR biomass and identified as Delftia tsuruhatensis and Pseudomonas aeruginosa. During incubation of the isolates, hydroquinone - a potentially toxic transformation product - was temporarily formed but further degraded and/or metabolized. These results suggest that the specific enrichment of a microbial consortium in an MBR operated at a high sludge age might be a promising strategy for post-treatment of WWTP effluents containing pharmaceuticals., (© 2010 Elsevier Ltd. All rights reserved.)

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/m(3) 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.

Pharmaceutical and personal care products, biocides and iodinated contrast media (ICM) are persistent compounds, which appear in ng to μg L(-1) in secondary effluents of sewage treatment plants (STPs). In this work, biogenic metals manganese oxides (BioMnOx) and bio-palladium (Bio-Pd) were applied in lab-scale membrane bioreactors (MBR) as oxidative and reductive technologies, respectively, to remove micropollutants from STP-effluent. From the 29 substances detected in the STP-effluent, 14 were eliminated in the BioMnOx-MBR: ibuprofen (>95%), naproxen (>95%), diuron (>94%), codeine (>93%), N-acetyl-sulfamethoxazole (92%), chlorophene (>89%), diclofenac (86%), mecoprop (81%), triclosan (>78%), clarithromycin, (75%), iohexol (72%), iopromide (68%), iomeprol (63%) and sulfamethoxazole (52%). The putative removal mechanisms were the chemical oxidation by BioMnOx and/or the biological removal by Pseudomonas putida and associated bacteria in the enriched biofilm. Yet, the removal rates (highest value: 2.6 μg diclofenac L(-1) d(-1)) need to improve by a factor 10 in order to be competitive with ozonation. ICM, persistent towards oxidative techniques, were successfully dehalogenated with a novel reductive technique using Bio-Pd as a nanosized catalyst in an MBR. Iomeprol, iopromide and iohexol were removed for >97% and the more recalcitrant diatrizoate for 90%. The conditions favorable for microbial H(2)-production enabling the charging of the Pd catalyst, were shown to be important for the removal of ICM. Overall, the results indicate that Mn oxide and Pd coupled to microbial catalysis offer novel potential for advanced water treatment., (© 2010 Elsevier Ltd. All rights reserved.)

In this study, the microbial community characteristics in continuous lab-scale anaerobic reactors were correlated to reactor functionality using the microbial resource management (MRM) approach. Two molecular techniques, denaturing gradient gel electrophoresis (DGGE) and terminal-restriction fragment length polymorphism (T-RFLP), were applied to analyze the bacterial and archaeal communities, and the results obtained have been compared. Clustering analyses showed a similar discrimination of samples with DGGE and T-RFLP data, with a clear separation between the meso- and thermophilic communities. Both techniques indicate that bacterial and mesophilic communities were richer and more even than archaeal and thermophilic communities, respectively. Remarkably, the community composition was highly dynamic for both Bacteria and Archaea, with a rate of change between 30% and 75% per 18 days, also in stable performing periods. A hypothesis to explain the latter in the context of the converging metabolism in anaerobic processes is proposed. Finally, a more even and diverse bacterial community was found to be statistically representative for a well-functioning reactor as evidenced by a low Ripley index and high biogas production.

Anaerobic oxidation of methane coupled to sulphate reduction (SR-AOM) prevents oceanic methane emissions and is considered as a major environmental process in the deep-sea sediments. To stimulate in vitro SR-AOM activity, we designed a (continuous) high-pressure bioreactor system. Fed-batch mode incubations showed that the elevated methane pressure stimulated SR-AOM activity: when the methane pressure increased from 1 to 4.5 to 8 MPa, the initial SR-AOM activity increased from 3.46 to 8.64 to 9.22 micromol sulphide production/g(dw)/day; the apparent affinity (K(m)) for methane was 37 mM. However, in each fed-batch mode incubation, there was an inhibitory effect observed after 2 days, probably due to the accumulation of sulphide and the increase of pH. When the reactor was operated in a continuous mode, the SR-AOM activity was constantly increasing within 18 days at both 1 and 8 MPa pressures., (Copyright 2009 Elsevier Ltd. All rights reserved.)

Three identical sequential batch reactors (SBRs) were each inoculated with sludge from a full-scale wastewater treatment plant (WWTP) treating a waste stream of different origin, i.e. a hospital, a meat processing company, and a municipal WWTP. The SBRs were run in parallel for 84 consecutive days to investigate whether the reactors would become more phylogenetically similar or stay separated concerning their functionality and microbial communities. Overall, the nitrification functionality was high throughout the experiment, and the size and structure of the sludge flocs were very similar. The total bacterial and ammonia-oxidizing bacterial (AOB) communities were analyzed by PCR-DGGE. Cluster analysis demonstrated very distinct bacterial communities in the three SBRs, not showing any trend becoming more similar. The carrying capacity, dynamics and functional organization of the communities were assessed by DGGE analysis and based on these patterns the range-weighted richness, moving window analysis, and constructing Pareto-Lorenz evenness distribution curves were calculated. Between the SBRs, highly comparable internal structure and dynamics of the AOB communities were observed, although they had only one AOB DGGE band in common. These observations indicate that community characteristics such as the extent of biodiversity and dynamics are more important indicators of good microbial functionality than the presence of certain specific species.

Trichloroethylene is a toxic and recalcitrant groundwater pollutant. Palladium nanoparticles bio-precipitated on Shewanella oneidensis were encapsulated in polyurethane, polyacrylamide, alginate, silica or coated on zeolites. The reactivity of these bio-Pd beads and zeolites was tested in batch experiments and trichloroethylene dechlorination followed first order reaction kinetics. The calculated k-values of the encapsulated catalysts were a factor of six lower compared to non-encapsulated bio-Pd. Bio-Pd, used as a catalyst, was able to dechlorinate 100 mgL(-1) trichloroethylene within a time period of 1h. The main reaction product was ethane; yet small levels of chlorinated intermediates were detected. Subsequently polyurethane cubes empowered with bio-Pd were implemented in a fixed bed reactor for the treatment of water containing trichloroethylene. The influent recycle configuration resulted in a cumulative removal of 98% after 22 h. The same reactor in a flow through configuration achieved removal rates up to 1059 mg trichloroethylene g Pd(-1)d(-1). This work showed that fixed bed reactors with bio-Pd polyurethane cubes can be instrumental for remediation of water contaminated with trichloroethylene.

Increasing concern about the fate of 17alpha-ethinylestradiol (EE2) in the environment stimulates the search for alternative methods for wastewater treatment plant (WWTP) effluent polishing. The aim of this study was to establish an innovative and effective biological removal technique for EE2 by means of a nitrifier enrichment culture (NEC) applied in a membrane bioreactor (MBR). In batch incubation tests, the microbial consortium was able to remove EE2 from both a synthetic minimal medium and WWTP effluent. A maximum EE2 removal rate of 9.0 microg EE2 g(-1)biomass-VSS h(-1) was achieved (>94% removal efficiency). Incubation of the heterotrophic bacteria isolated from the NEC did not result in a significant EE2 removal, indicating the importance of nitrification as driving force in the mechanism. Application of the NEC in a MBR to treat a synthetic influent with an EE2 concentration of 83 ng EE2 L(-1) resulted in a removal efficiency of 99% (loading rates up to 208 ng EE2 L(-1)d(-1); membrane flux rate: 6.9 L m(-2) h(-1)). Simultaneously, complete nitrification was achieved at an optimal ammonium influent concentration of 1.0 mg NH(4)(+)-N L(-1). This minimal NH(4)(+)-N input is very advantageous for effluent polishing since the concomitant effluent nitrate concentrations will be low as well and it offers opportunities for the nitrifying MBR as a promising add-on technology for WWTP effluent polishing.

Trichloroethylene (TCE) is a toxic and recalcitrant groundwater pollutant. An innovative technology using microbial produced Pd(0) nanoparticles for the remediation of TCE contaminated groundwater was developed. The nanoscale bio-Pd particles were precipitated on the biomass of Shewanella oneidensis and hydrogen gas, formate, or formic acid were used as hydrogen donors. Ethane turned out to be the only organic degradation product and no intermediate chlorinated reaction products were detected. Subsequently bio-Pd was implemented in a plate membrane reactor (MR) for the treatment of water containing TCE. In a continuous MR system, containing 50 mg L(-1) bio-Pd, removal rates up to 2,515 mg TCE day(-1) g(-1) Pd were achieved with H(2) gas as hydrogen donor. The measured chloride mass balance confirmed the removal rates. This work shows that a complete, efficient and rapid removal of TCE was achieved with bio-Pd and that a MR system containing bio-Pd and supplied with hydrogen gas offers an alternative for the current remediation technologies of water contaminated with TCE.

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 x day, achieved during the aerobic days of the cycles. Denaturing gradientgel 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.

The autotrophic two-species biofilm from the packed bed reactor of a life-support system, containing Nitrosomonas europaea ATCC 19718 and Nitrobacter winogradskyi ATCC 25391, was analysed after 4.8 years of continuous operation performing complete nitrification. Real-time quantitative polymerase chain reaction (Q-PCR) was used to quantify N. europaea and N. winogradskyi along the vertical axis of the reactor, revealing a spatial segregation of N. europaea and N. winogradskyi. The main parameters influencing the spatial segregation of both nitrifiers along the bed were assessed through a multi-species one-dimensional biofilm model generated with AQUASIM software. The factor that contributed the most to this distribution profile was a small deviation from the flow pattern of a perfectly mixed tank towards plug-flow. The results indicate that the model can estimate the impact of specific biofilm parameters and predict the nitrification efficiency and population dynamics of a multispecies biofilm.

A sequential batch reactor (SBR) and a membrane bioreactor (MBR) were inoculated with the same sludge from a municipal wastewater treatment plant, supplemented with ammonium, and operated in parallel for 84 days. It was investigated whether the functional stability of the nitrification process corresponded with a static ammonia-oxidizing bacterial (AOB) community. The SBR provided complete nitrification during nearly the whole experimental run, whereas the MBR showed a buildup of 0 to 2 mg nitrite-N liter(-1) from day 45 until day 84. Based on the denaturing gradient gel electrophoresis profiles, two novel approaches were introduced to characterize and quantify the community dynamics and interspecies abundance ratios: (i) the rate of change [Delta(t)((week))] parameter and (ii) the Pareto-Lorenz curve distribution pattern. During the whole sampling period, it was observed that neither of the reactor types maintained a static microbial community and that the SBR evolved more gradually than the MBR, particularly with respect to AOB (i.e., average weekly community changes of 12.6% +/- 5.2% for the SBR and 24.6% +/- 14.3% for the MBR). Based on the Pareto-Lorenz curves, it was observed that only a small group of AOB species played a numerically dominant role in the nitritation of both reactors, and this was true especially for the MBR. The remaining less dominant species were speculated to constitute a reserve of AOB which can proliferate to replace the dominant species. The value of these parameters in terms of tools to assist the operation of activated-sludge systems is discussed.

Connecting several microbial fuel cell (MFC) units in series or parallel can increase voltage and current; the effect on the microbial electricity generation was as yet unknown. Six individual continuous MFC units in a stacked configuration produced a maximum hourly averaged power output of 258 W m(-3) using a hexacyanoferrate cathode. The connection of the 6 MFC units in series and parallel enabled an increase of the voltages (2.02 V at 228 W m(-3)) and the currents (255 mA at 248 W m(-3)), while retaining high power outputs. During the connection in series, the individual MFC voltages diverged due to microbial limitations at increasing currents. With time, the initial microbial community decreased in diversity and Gram-positive species became dominant. The shift of the microbial community accompanied a tripling of the short time power output of the individual MFCs from 73 W m(-3) to 275 W m(-3), a decrease of the mass transfer limitations and a lowering of the MFC internal resistance from 6.5 +/- 1.0 to 3.9 +/- 0.5 omega. This study demonstrates a clear relation between the electrochemical performance and the microbial composition of MFCs and further substantiates the potential to generate useful energy by means of MFCs.

Bioaugmentation of bioreactors focuses on the removal of xenobiotics, with little attention typically paid to the recovery of disrupted reactor functions such as ammonium-nitrogen removal. Chloroanilines are widely used in industry as a precursor to a variety of products and are occasionally released into wastewater streams. This work evaluated the effects on activated-sludge reactor functions of a 3-chloroaniline (3-CA) pulse and bioaugmentation by inoculation with the 3-CA-degrading strain Comamonas testosteroni I2 gfp. Changes in functions such as nitrification, carbon removal, and sludge compaction were studied in relation to the sludge community structure, in particular the nitrifying populations. Denaturing gradient gel electrophoresis (DGGE), real-time PCR, and fluorescent in situ hybridization (FISH) were used to characterize and enumerate the ammonia-oxidizing microbial community immediately after a 3-CA shock load. Two days after the 3-CA shock, ammonium accumulated, and the nitrification activity did not recover over a 12-day period in the nonbioaugmented reactors. In contrast, nitrification in the bioaugmented reactor started to recover on day 4. The DGGE patterns and the FISH and real-time PCR data showed that the ammonia-oxidizing microbial community of the bioaugmented reactor recovered in structure, activity, and abundance, while the number of ribosomes of the ammonia oxidizers in the nonbioaugmented reactor decreased drastically and the community composition changed and did not recover. The settleability of the activated sludge was negatively influenced by the 3-CA addition, with the sludge volume index increasing by a factor of 2.3. Two days after the 3-CA shock in the nonbioaugmented reactor, chemical oxygen demand (COD) removal efficiency decreased by 36% but recovered fully by day 4. In contrast, in the bioaugmented reactor, no decrease of the COD removal efficiency was observed. This study demonstrates that bioaugmentation of wastewater reactors to accelerate the degradation of toxic chlorinated organics such as 3-CA protected the nitrifying bacterial community, thereby allowing faster recovery from toxic shocks.

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 I2, 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 I2gfp 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 I2 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.

Effluents of anaerobic digesters are an underestimated source of greenhouse gases, as they are often saturated with methane. A post-treatment with methane-oxidizing bacterial consortia could mitigate diffuse emissions at such sites. Semi-continuously fed stirred reactors were used as model systems to characterize the influence of the key parameters on the activity of these mixed methanotrophic communities. The addition of 140 mg L−1 NH–N had no significant influence on the activity nor did a temperature increase from 28°C to 35°C. On the other hand, addition of 0.64 mg L−1 of copper(II) increased the methane removal rate by a factor of 1.5 to 1.7 since the activity of particulate methane monooxygenase was enhanced. The influence of different concentrations of NaCl was also tested, as effluents of anaerobic digesters often contain salt levels up to 10 g NaCl L−1. At a concentration of 11 g NaCl L−1, almost no methane-oxidizing activity was observed in the reactors without copper addition. Yet, reactors with copper addition exhibited a sustained activity in the presence of NaCl. A colorimetric test based on naphthalene oxidation showed that soluble methane monooxygenase was inhibited by copper, suggesting that the particulate methane monooxygenase was the active enzyme and thus more salt resistant. The results obtained demonstrate that the treatment of methane-saturated effluents, even those with increased ammonium (up to 140 mg L−1 NH–N) and salt levels, can be mitigated by implementation of methane-oxidizing microbial consortia. [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]

So far, researchers investigated microbially induced CaCO 3 precipitation (MICP) for soil reinforcement, self-repairing concrete and Ca 2+ removal from industrial waste streams. Reported MICP yields were mainly achieved under nutrient-rich conditions. However, creating the tested nutrient-rich conditions in intended applications is both an economical and a practical issue. Therefore, investigation of MICP in more realistic conditions is necessary. This study presents optimization of MICP through denitrification in minimal nutrient conditions. To optimize their MICP performances, we isolated two strains, Pseudomonas aeruginosa and Diaphorobacter nitroreducens , by following an application oriented selection procedure. Upon performance optimization, in 2 days, D. nitroreducens and P. aeruginosa precipitated 14.1 and 18.9 g CaCO 3 /g NO 3 -N, respectively. Repetitive CaCO 3 precipitation was also achieved from a single inoculum in both 2 days and 3 weeks intervals. Selected strains and the process were further evaluated for three MICP applications: (1) Ca 2+ removal from paper mill wastewater (2) soil reinforcement, (3) crack repair in concrete. Overall, denitrification was found to be an effective process to remove Ca 2+ from paper mill wastewater. P. aeruginosa and D. nitroreducens could be introduced as potential candidates for soil and concrete applications due to their enhanced precipitation yields, resilience and performance under minimal nutrient conditions. [ABSTRACT FROM AUTHOR]

Nitrification and denitrification, the key biological processes for thermophilic nitrogen removal, have separately been established in bioreactors at 50 °C. A well-characterized set of kinetic parameters is essential to integrate these processes while safeguarding the autotrophs performing nitrification. Knowledge on thermophilic nitrifying kinetics is restricted to isolated or highly enriched batch cultures, which do not represent bioreactor conditions. This study characterized the stoichiometry and kinetics of two thermophilic (50 °C) nitrifying communities. The most abundant ammonia oxidizing archaea (AOA) were related to the Nitrososphaera genus, clustering relatively far from known species Nitrososphaera gargensis (95.5% 16S rRNA gene sequence identity). The most abundant nitrite oxidizing bacteria (NOB) were related to Nitrospira calida (97% 16S rRNA gene sequence identity). The nitrification biomass yield was 0.20–0.24 g VSS g−1 N, resulting mainly from a high AOA yield (0.16–0.20 g VSS g−1 N), which was reflected in a high AOA abundance in the community (57–76%) compared to NOB (5–11%). Batch-wise determination of decay rates (AOA: 0.23–0.29 d−1; NOB: 0.32–0.43 d−1) rendered an overestimation compared to in situ estimations of overall decay rate (0.026–0.078 d−1). Possibly, the inactivation rate rather than the actual decay rate was determined in batch experiments. Maximum growth rates of AOA and NOB were 0.12–0.15 d−1 and 0.13–0.33 d−1 respectively. NOB were susceptible to nitrite, opening up opportunities for shortcut nitrogen removal. However, NOB had a similar growth rate and oxygen affinity (0.15–0.55 mg O 2 L−1) as AOA and were resilient towards free ammonia (IC 50 > 16 mg NH 3 -N L−1). This might complicate NOB outselection using common practices to establish shortcut nitrogen removal (SRT control; aeration control; free ammonia shocks). Overall, the obtained insights can assist in integrating thermophilic conversions and facilitate single-sludge nitrification/denitrification. Image 1 • Thermophilic nitrification yield was high, resulting mainly from a high AOA yield. • AOA belonged to the Nitrososphaera genus, NOB to Nitrospira calida. • Batch-wise determined decay rates were overestimated compared to in situ estimation. • NOB kinetics and high FA tolerance might complicate thermophilic NOB outselection. • Higher maximum temperature for AOA activity retention (55 °C) vs NOB (50 °C). [ABSTRACT FROM AUTHOR]

Cost-efficient biological treatment of warm nitrogenous wastewaters requires the development of thermophilic nitrogen removal processes. Only one thermophilic nitrifying bioreactor was described so far, achieving 200 mg N L −1 d −1 after more than 300 days of enrichment from compost samples. From the practical point of view in which existing plants would be upgraded, however, a more time-efficient development strategy based on mesophilic nitrifying sludge is preferred. This study evaluated the adaptive capacities of mesophilic nitrifying sludge for two linear temperature increase patterns (non-oscillating vs. oscillating), two different slopes (0.25 vs. 0.08 °C d −1 ) and two different reactor types (floc vs. biofilm growth). The oscillating temperature pattern (0.25 °C d −1 ) and the moving bed biofilm reactor (0.08 °C d −1 ) could not reach nitrification at temperatures higher than 46 °C. However, nitrification rates up to 800 mg N L −1 d −1 and 150 mg N g −1 volatile suspended solids d −1 were achieved at a temperature as high as 49 °C by imposing the slowest linear temperature increase to floccular sludge. Microbial community analysis revealed that this successful transition was related with a shift in ammonium oxidizing archaea dominating ammonia oxidizing bacteria, while for nitrite oxidation Nitrospira spp. was constantly more abundant than Nitrobacter spp.. This observation was accompanied with an increase in observed sludge yield and a shift in maximal optimum temperature, determined with ex-situ temperature sensitivity measurements, predicting an upcoming reactor failure at higher temperature. Overall, this study achieved nitrification at 49 °C within 150 days by gradual adaptation of mesophilic sludge, and showed that ex-situ temperature sensitivity screening can be used to monitor and steer the transition process. [ABSTRACT FROM AUTHOR]

Bio-refineries produce large volumes of waste streams with high organic content, which are potentially interesting for further processing. Anaerobic digestion (AD) can be a key technology for treatment of these sidestreams, such as molasses. However, the high concentration of salts in molasses can cause inhibition of methanogenesis. In this research, concentrated and diluted molasses were subjected to biomethanation in two types of submerged anaerobic membrane bioreactors (AnMBRs): one with biogas recirculation and one with a vibrating membrane. Both reactors were compared in terms of methane production and membrane fouling. Biogas recirculation seemed to be a good way to avoid membrane fouling, while the trans membrane pressures in the vibrating MBR increased over time, due to cake layer formation and the absence of a mixing system. Stable methane production, up to 2.05 L L−1 d−1 and a concomitant COD removal of 94.4%, was obtained only when diluted molasses were used, since concentrated molasses caused a decrease in methane production and an increase in volatile fatty acids (VFA), indicating an inhibiting effect of concentrated molasses on AD. Real-time PCR results revealed a clear dominance of Methanosaetaceae over Methanosarcinaceae as the main acetoclastic methanogens in both AnMBRs. [ABSTRACT FROM AUTHOR]

Abstract: In this study we validated the use of 24 square deepwell microtiterplates to screen large libraries of metabolically engineered strains by investigating the optimization of succinate production. Wild type E. coli MG1655 and 11 derived mutants were physiologically evaluated by growth in 24 deepwell MTPs and 2L benchtop bioreactors. Growth parameters, product yields and byproduct formation were determined for all mutants. The results show that similar average values and standard deviations for these parameters were obtained. Especially a high correlation was noticed for the acetate byproduct yield and the succinate production rate. For these parameters there was no significant difference for 8 out of 12 strains between MTPs and 2L bioreactors. However a lower maximum growth rate was observed in 2L reactors as opposed to 24 deepwell plates for 9 out of 12 mutants which could be linked to the higher amount of dead cells in the benchtop bioreactors (12% vs. 2% in MTPs). Finally, a cluster-based approach was used to select good producer strains, i.e. strains with a high succinate yield and succinate production rate. Bad, intermediate and good producer strains were clustered in the same groups for MTPs and benchtop bioreactors for 11 out of the 12 investigated strains. [ABSTRACT FROM AUTHOR]