Data on the influence of temperature on the growth of Escherichia coli in a minimal medium containing glucose as the sole carbon source for the joint computation of growth yields and rates at each temperature from 27 to 45°C.

Temperature is a key factor influencing microbial growth rates and yields. In literature, the influence of temperature on growth is studied either on yields or rates but not both at the same time. Moreover, studies often report the influence of a specific set of temperatures using rich culture media containing complex ingredients (such as yeast extract) which chemical composition cannot be precisely specified. Here, we present a complete dataset for the growth of Escherichia coli K12 NCM3722 strain in a minimal medium containing glucose as the sole energy and carbon source for the computation of growth yields and rates at each temperature from 27 to 45°C. For this purpose, we monitored the growth of E. coli by automated optical density (OD) measurements in a thermostated microplate reader. At each temperature full OD curves were reported for 28 to 40 microbial cultures growing in parallel wells. Additionally, a correlation was established between OD values and the dry mass of E. coli cultures. For that, 21 dilutions were prepared from triplicate cultures and optical density was measured in parallel with the microplate reader (ODmicroplate) and a UV-Vis spectrophotometer (ODUV-vis) and correlated to duplicate dry biomass measurements. The correlation was used to compute growth yields in terms of dry biomass.

Diversity of novel archaeal viruses infecting methanogens discovered through coupling of stable isotope probing and metagenomics.

Diversity of viruses infecting non-extremophilic archaea has been grossly understudied. This is particularly the case for viruses infecting methanogenic archaea, key players in the global carbon biogeochemical cycle. Only a dozen of methanogenic archaeal viruses have been isolated so far. In the present study, we implemented an original coupling between stable isotope probing and complementary shotgun metagenomic analyses to identify viruses of methanogens involved in the bioconversion of formate, which was used as the sole carbon source in batch anaerobic digestion microcosms. Under our experimental conditions, the microcosms were dominated by methanogens belonging to the order Methanobacteriales (Methanobacterium and Methanobrevibacter genera). Metagenomic analyses yielded several previously uncharacterized viral genomes, including a complete genome of a head-tailed virus (class Caudoviricetes, proposed family Speroviridae, Methanobacterium host) and several near-complete genomes of spindle-shaped viruses. The two groups of viruses are predicted to infect methanogens of the Methanobacterium and Methanosarcina genera and represent two new virus families. The metagenomics results are in good agreement with the electron microscopy observations, which revealed the dominance of head-tailed virus-like particles and the presence of spindle-shaped particles. The present study significantly expands the knowledge on the viral diversity of viruses of methanogens.

Systematic and quantitative analysis of two decades of anodic wastewater treatment in bioelectrochemical reactors.

Wastewater treatment is generally performed using energy-intensive processes, such as activated sludge. Improving energy efficiency has become one of the main challenges for next-generation wastewater treatment plants. Bioelectrochemical systems (BES) have been attracting attention because they take advantage of the chemical energy contained in wastewater while enabling the valorization of effluents: either with electrical energy (microbial fuel cells) or with useful chemicals (microbial electrolysis cells). Bioelectrochemical wastewater treatment has been under investigation since the early 2000s and is now the subject of an abundant literature, which is most frequently focused on anodic COD removal. Comparing results obtained in different studies is particularly difficult with BES, because many different parameters (effluent characteristics, inoculation, design, and operation) may interact and because using real effluents results in high variability. To address this issue, data were retrieved from 1,073 articles that were selected objectively and with transparency. This systematic review evaluates the potential of anodic wastewater treatment, based on 4,579 experimental observations. Overall, BES has already shown satisfactory treatment capacity, with a median chemical oxygen demand removal of 72%. However, the median coulombic efficiency was only 18%, increasing this parameter offers the greatest opportunity for BES improvement.

Gradual development of ammonia-induced syntrophic acetate-oxidizing activities under mesophilic and thermophilic conditions quantitatively tracked using multiple isotopic approaches.

Insights into microbiota adaptation to increased ammonia stress, and identification of indicator microorganisms can help to optimize the operation of anaerobic digesters. To identify microbial indicators and investigate their metabolic contribution to acetoclastic methanogenesis (AM), syntrophic acetate oxidation (SAO) or hydrogenotrophic methanogenesis (HM), 40 anaerobic batch reactors fed with acetate of 110 mmol/L were set up at NH4+-N concentrations of 0.14 g/L, 5.00 g/L or 7.00 g/L, inoculated with thermophilic or mesophilic microbiota with or without pre-exposure to ammonia stress. Four stable carbon isotope probing approaches were applied in parallel, with [1,2-13C]-CH3COOH, [2-13C]-CH3COOH, [13C]NaHCO3 or non-labeled CH3COOH used individually. The last three approaches were used to quantify the methanogenic pathways by tracking labeled 13C or natural 13C signatures in the resulting CH4 and CO2, and consistently detected the dynamic transition of dominant pathways from AM to SAO-HM under ammonia stress. Results of quantitative PCR and fluorescence in-situ hybridization illustrated the procedure, acetotrophic methanogens being outcompeted by acetate-oxidizing syntrophs. The first and last isotope-labeling approaches were designed to probe the active acetate-mineralizing microbes with DNA-SIP. Known acetate-oxidizing bacteria like Syntrophaceticus and Tepidanaerobacter, as well as novel members of Pseudomonas, Bacillus and Symbiobacteraceae were detected, with Methanoculleus as the predominant H2/CO2-utilizing partner. Using NanoSIMS, some bacterial cells were observed to be fixing CO2 from [13C]NaHCO3. In this study, Methanosaeta was only active with ammonia < 200 mg-N/L; the syntrophs catalyzing SAO-HM started to compete with AM-conducting Methanosarcina at intermediate concentrations of ammonia, i.e. 200-500 mg-N/L, and outcompeted the acetotrophic methanogens with ammonia > 500 mg-N/L. Under ammonia stress, diverse known and novel microbial taxa were involved in acetate mineralization, comparable with those identified in previous studies.

Electrochemical analysis of a microbial electrochemical snorkel in laboratory and constructed wetlands.

Microbial electrochemical snorkel (MES) is a short-circuited microbial fuel cell applicable to water treatment that does not produce energy but requires lower cost for its implementation. Few reports have already described its water treatment capabilities but no deeper electrochemical analysis were yet performed. We tested various materials (iron, stainless steel and porous graphite) and configurations of snorkel in order to better understand the rules that will control in a wetland the mixed potential of this self-powered system. We designed a model snorkel that was studied in laboratory and on the field. We confirmed the development of MES by identifying anodic and cathodic parts, by measuring the current between them and by analyzing microbial ecology in laboratory and field experiments. An important application is denitrification of surface water. Here we discuss the influence of nitrate on its electrochemical response and denitrification performances. Introducing nitrate caused the increase of the mixed potential of MES and of current at a potential value relatively more positive than for nitrate-reducing biocathodes described in the literature. The major criteria for promoting application of MES in artificial wetland dedicated to mitigation of non-point source nitrate pollution from agricultural water are considered.

Denitrifying bio-cathodes developed from constructed wetland sediments exhibit electroactive nitrate reducing biofilms dominated by the genera Azoarcus and Pontibacter.

To limit the nitrate contamination of ground and surface water, stimulation of denitrification by electrochemical approach is an innovative way to be explored. Two nitrate reducing bio-cathodes were developed under constant polarization (-0.5 V vs SCE) using sediments and water from a constructed wetland (Rampillon, Seine-et-Marne, France). The bio-cathodes responded to nitrate addition on chronoamperometry through an increase of the reductive current. The denitrification efficiency of the pilots increased by 47% compared to the negative controls without electrodes after polarization. 16S rRNA gene sequencing of the biofilms and sediments evidenced the significant and discriminating presence of the Azoarcus and Pontibacter genera in the biofilms from biocathodes active for nitrate reduction. Our study shows the possibility to promote the development of efficient Azoarcus-dominated biocathodes from freshwater sediment to enhance nitrate removal from surface waters.

Insights from Microbial Transition State Theory on Monod's Affinity Constant.

Microbial transition state theory (MTS) offers a theoretically explicit mathematical model for substrate limited microbial growth. By considering a first order approximation of the MTS equation one recovers the well-known Monod's expression for growth, which was regarded as a purely empirical function. The harvest volume of a cell as defined in MTS theory can then be related to the affinity concept, giving a new physical interpretation to it, and a new way to determine its value. Consequences of such a relationship are discussed.

Biorefinery for heterogeneous organic waste using microbial electrochemical technology.

Environmental biorefineries aim to produce biofuels and platform biomolecules from organic waste. To this end, microbial electrochemical technologies theoretically allow controlled microbial electrosynthesis (MES) of organic molecules to be coupled to oxidation of waste. Here, we provide a first proof of concept and a robust operation strategy for MES in a microbial electrolysis cell (MEC) fed with biowaste hydrolysates. This strategy allowed stable operation at 5 A/m2 for more than three months in a labscale reactor. We report a two to four-fold reduction in power consumption compared to microbial electrosynthesis with water oxidation at the anode. The bioelectrochemical characterizations of the cells were used to compute energy and matter balances for biorefinery scenarios in which anaerobic digestion (AD) provides the electricity and CO2 required for the MEC. Calculations shows that up to 22% of electrons (or COD) from waste may be converted to organic products in the AD-MEC process.

Comparative metatranscriptomic analysis of anaerobic digesters treating anionic surfactant contaminated wastewater.

Three distinct biological reactors fed with synthetic medium (UASB_Control), synthetic medium and linear alkylbenzene sulfonate (LAS; UASB_SL), and real laundry wastewater (UASB_LW) were compared using a metatranscriptomic approach to determine putative bioindicator genes and taxonomies associated to all steps of anaerobic LAS biodegradation pathway. A homemade bioinformatics pipeline combined with an R workflow was developed to perform the RNAseq data analysis. UASB_SL and UASB_LW showed similar values of LAS biological degradation (~47%) and removal (53-55%). Rarefaction analysis revealed that 1-2 million reads were sufficient to access the whole functional capacity. In the first step of LAS biodegradation pathway, fumarate reductase subunit C was detected and taxonomically assigned to the genus Syntrophobacter (0.002% - UASB_SL; 0.0015% - UASB_LW; not detected - UASB_Control). In the second step, many enzymes related to beta-oxidation were observed and most of them with low relative abundance in UASB Control and taxonomically related with Smithella, Acinetobacter and Syntrophorhabdus. For the ring cleavage step, the abundance of 6 OCH CoA hydrolase putative gene was ten times higher in UASB_SL and UASB_LW when compared to UASB_Control, and assigned to Desulfomonile and Syntrophorhabdus. Finally, the adenylylsulfate reductase, taxonomically related with Desulfovibrio and Desulfomonile, was observed in the desulfonation step with the highest relative abundance in UASB_LW.

Consistent microbial dynamics and functional community patterns derived from first principles.

Microbial communities are key engines that drive earth's biogeochemical cycles. However, existing ecosystem models have only limited ability to predict microbial dynamics and require the calibration of multiple population-specific empirical equations. In contrast, we build on a new kinetic "Microbial Transition State" (MTS) theory of growth derived from first principles. We show how the theory coupled to simple mass and energy balance calculations provides a framework with intrinsically important qualitative properties to model microbial community dynamics. We first show how the theory can simultaneously account for the influence of all the resources needed for growth (electron donor, acceptor, and nutrients) while still producing consistent dynamics that fulfill the Liebig rule of a single limiting substrate. We also show consistent patterns of energy-dependent microbial successions in mixed culture without the need for calibration of population-specific parameters. We then show how this approach can be used to model a simplified activated sludge community. To this end, we compare MTS-derived dynamics with those of a widely used activated sludge model and show that similar growth yields and overall dynamics can be obtained using two parameters instead of twelve. This new kinetic theory of growth grounded by a set of generic physical principles parsimoniously gives rise to consistent microbial population and community dynamics, thereby paving the way for the development of a new class of more predictive microbial ecosystem models.

Life cycle assessment of a bioelectrochemical system as a new technological platform for biosuccinic acid production from waste.

Waste management is a key environmental and socio-economic issue. Environmental concerns are encouraging the use of alternative resources and lower emissions to air, water and soil. Innovative technologies to deal with waste recovery that produce marketable bioproducts are emerging. Bioelectrochemical synthesis systems (BESs) are based on the primary principle of transforming organic waste into added-value products using microorganisms to catalyse chemical reactions. This technology is at the core of a research project called BIORARE (BIoelectrosynthesis for ORganic wAste bioREfinery), an interdisciplinary project that aims to use anaerobic digestion as a supply chain to feed a BES and produce target biomolecules. This technology needs to be driven by environmental strategies. Life Cycle Assessment (LCA) was used to evaluate the BIORARE concept based on expert opinion and prior experiments for the production of biosuccinic acid and waste management. A multidisciplinary approach based on biochemistry and process engineering expertise was used to collect the inventory data. The BES design and the two-step anaerobic digestion process have many potential impacts on air pollution or ecotoxicity-related categories. The comparison of the BIORARE concept with conventional fermentation processes and a water-fed BES technology demonstrated the environmental benefit resulting from the use of both the BES technology and a waste-based substrate as input thus supporting the BIORARE concept. Some trade-offs among the impact categories were identified but led to options to improve the concept. BES design and synergy management may improve the environmental performance of the BIORARE concept.

Improving anaerobic digestion with support media: Mitigation of ammonia inhibition and effect on microbial communities.

This study aimed at providing a better understanding of the influence of support media (10g/L of zeolites, 10g/L of activated carbons, and 1g/L of chitosan) on key phylotypes steering anaerobic digestion (AD) performance in presence of 19g/L of Total Ammonia Nitrogen (TAN) within batch digesters. Support media did not influence TAN concentration. However, both zeolites and activated carbon 1 reduced methanization lag phase by 47% and 25%, respectively. By contrast, activated carbon 2 and chitosan led to an increase of methanization lag phase by 51% and 32%, respectively. 16S rRNA gene sequencing revealed that zeolites preserved Methanosarcina and enhanced Methanobacterium. In presence of activated carbon 1, Methanoculleus, became predominant earlier than without support while chitosan and activated carbon 2 limited its implantation. This study highlighted potentialities to use supports to enhance AD stability under extreme TAN concentration and evidenced their specific influence on the microbiota composition.

Whole Proteome Analyses on Ruminiclostridium cellulolyticum Show a Modulation of the Cellulolysis Machinery in Response to Cellulosic Materials with Subtle Differences in Chemical and Structural Properties.

Lignocellulosic materials from municipal solid waste emerge as attractive resources for anaerobic digestion biorefinery. To increase the knowledge required for establishing efficient bioprocesses, dynamics of batch fermentation by the cellulolytic bacterium Ruminiclostridium cellulolyticum were compared using three cellulosic materials, paper handkerchief, cotton discs and Whatman filter paper. Fermentation of paper handkerchief occurred the fastest and resulted in a specific metabolic profile: it resulted in the lowest acetate-to-lactate and acetate-to-ethanol ratios. By shotgun proteomic analyses of paper handkerchief and Whatman paper incubations, 151 proteins with significantly different levels were detected, including 20 of the 65 cellulosomal components, 8 non-cellulosomal CAZymes and 44 distinct extracytoplasmic proteins. Consistent with the specific metabolic profile observed, many enzymes from the central carbon catabolic pathways had higher levels in paper handkerchief incubations. Among the quantified CAZymes and cellulosomal components, 10 endoglucanases mainly from the GH9 families and 7 other cellulosomal subunits had lower levels in paper handkerchief incubations. An in-depth characterization of the materials used showed that the lower levels of endoglucanases in paper handkerchief incubations could hypothetically result from its lower crystallinity index (50%) and degree of polymerization (970). By contrast, the higher hemicellulose rate in paper handkerchief (13.87%) did not result in the enhanced expression of enzyme with xylanase as primary activity, including enzymes from the "xyl-doc" cluster. It suggests the absence, in this material, of molecular structures that specifically lead to xylanase induction. The integrated approach developed in this work shows that subtle differences among cellulosic materials regarding chemical and structural characteristics have significant effects on expressed bacterial functions, in particular the cellulolysis machinery, resulting in different metabolic patterns and degradation dynamics.

New insights into the key microbial phylotypes of anaerobic sludge digesters under different operational conditions.

Analyses on bacterial, archaeal communities at family level and methane-production metabolism were conducted in thirteen full-scale and pilot-scale anaerobic sludge digesters. These digesters were operated at different conditions regarding solids concentration, sludge retention time, organic loading rate and feedstock composition, seeking to optimize digester capacity. Correlations between process parameters and identified microbial phylotypes were evaluated based on relative abundance of these phylotypes determined by Quantitative PCR and 16S rDNA amplicon sequencing. Results showed that, Total Solids concentration (TS), among the evaluated operational parameters, demonstrated the most positive correlation with chemical parameters (including NH3 and VFAs) and significant impact on the abundance of key microbial phylotypes regardless of other factors. Digesters were grouped into 'Higher-TS' with higher stress (TS > 44 g/L, NH3 > 90 mg/L, VFAs > 300 mg/L) and 'Lower-TS' under easier status (TS ≤ 44 g/L, NH3 < 120 mg/L, VFAs < 525 mg/L) in this study. We identified the key microbial phylotypes, i.e. the most abundant and discriminating populations, in 'Higher-TS' digesters with high biogas production rate, which were the class Clostridia, the family Methanosarcinaceae and the order Methanobacteriales. Thermoanaerobacteraceae and Syntrophomonadaceae were identified as key families of Clostridia. Methane was produced both from acetoclastic and hydrogenotrophic methanogenesis. By contrast, in 'Higher-TS' digesters with low biogas production rate, the classes Alpha-, Beta- and Gamma-proteobacteria were detected in higher percentages, of which Rhodobacteraceae, Comamonadaceae and Xanthomonadaceae were the most abundant families respectively, and Methanomicrobiales was the prevailing methanogen order. Consistently, hydrogenotrophic pathway was predominant for methanogenesis, indicating existence of syntrophic acetate oxidation in such 'high-stress', low biogas production rate digesters. These microbial phylotypes were therefore considered to be associated to 'Higher-TS' operation. In 'Lower-TS' digesters, the abundance of the class Delta-proteobacteria, the families Anaerolineaceae, Rikenellaceae, Candidatus Cloacamonas and Methanosaetaceae was obviously higher compared with those in 'Higher-TS' digesters, which were thus considered to be marker phylotypes of easy status. The influence of TS and NH3 on the microbiome should be considered when a 'TS-increasing' strategy is applied to increase digester capacity.

Community shifts within anaerobic digestion microbiota facing phenol inhibition: Towards early warning microbial indicators?

Performance stability is a key operational issue for anaerobic digestion (AD) and phenolic compounds are regularly mentioned as a major cause of digester failures. To get more insights into AD microbiota response to a wide range of inhibition levels, anaerobic batch toxicity assays were conducted with ten phenol concentrations up to 5.00 g/L. Final AD performance was not impaired up to 1.00 g/L. However, progressive shifts in microbial community structure were detected from 0.50 g/L. The methanogenic function was maintained along with increasing initial phenol concentrations up to 2.00 g/L thanks to the emergence of genus Methanoculleus at the expense of Methanosarcina. Within syntrophic populations, family Syntrophomonadaceae proportion was gradually reduced by phenol while Synergistaceae gained in importance in the microbiome. Moreover, at 2.00 g/L, the relative abundance of families belonging to order Clostridiales dropped, leading to the predominance of populations assigned to order Bacteroidales even though it did not prevent final AD performance deterioration. It illustrates the high level of adaptability of archaeal and bacterial communities and suggests the possibility of determining early warning microbial indicators associated with phenol inhibition.

Penta- and 2,4,6-tri-chlorophenol biodegradation during municipal solid waste anaerobic digestion.

In this study isotopic tracing using (13)C labelled pentachlorophenol (PCP) and 2,4,6-trichlorophenol (2,4,6-TCP) is proposed as a tool to distinguish the loss of PCP and 2,4,6-TCP due to biodegradation from other physical processes. This isotopic approach was applied to accurately assess in situ PCP and 2,4,6-TCP degradation under methanogenic conditions in several microcosms made up of household waste. These microcosms were incubated in anaerobic conditions at 35°C (mesophilic) and 55°C (thermophilic) without agitation. The volume of biogas produced (CH4 and CO2), was followed for a period of 130 days. At this stage of stable methanogenesis, (13)C6-PCP and (13)C6-2,4,6-TCP were introduced anaerobically in microcosms and its monitoring at mesophilic and thermophilic conditions was performed in parallel by gas chromatography mass spectrometry (GC-MS) and gas chromatography isotope-ratio mass spectrometry (GC-IRMS). This study proved the almost total dechlorination of bioavailable PCP and 2,4,6-TCP into 4-CP at 35°C. Nevertheless, high rate adsorption in particular materials of the two compounds was observed. Furthermore, Carbon-13 Nuclear Magnetic Resonance ((13)C-NMR) Spectroscopy analysis of (13)C labelled 2,4,6-TCP mesophilic incubations showed the partial mineralization of 4-CP at 35°C to acetate and then to HCO(3-). Consequently, NMR results confirm the biogas isotopic results indicating the mineralization of (13)C labelled 2,4,6-TCP into (13)C (CH4 and CO2). Concerning (13)C labelled PCP mesophilic incubations, the isotopic composition of the biogas still natural until the day 262. In contrast, no dechlorination was observed at 55°C. Thus PCP and 2,4,6-TCP were persistent in thermophilic conditions.

Biocathodes reducing oxygen at high potential select biofilms dominated by Ectothiorhodospiraceae populations harboring a specific association of genes.

Biocathodes polarized at high potential are promising for enhancing Microbial Fuel Cell performances but the microbes and genes involved remain poorly documented. Here, two sets of five oxygen-reducing biocathodes were formed at two potentials (-0.4V and +0.1V vs. saturated calomel electrode) and analyzed combining electrochemical and metagenomic approaches. Slower start-up but higher current densities were observed at high potential and a distinctive peak increasing over time was recorded on cyclic voltamogramms, suggesting the growth of oxygen reducing microbes. 16S pyrotag sequencing showed the enrichment of two operational taxonomic units (OTUs) affiliated to Ectothiorodospiraceae on high potential electrodes with the best performances. Shotgun metagenome sequencing and a newly developed method for the identification of Taxon Specific Gene Annotations (TSGA) revealed Ectothiorhodospiraceae specific genes possibly involved in electron transfer and in autotrophic growth. These results give interesting insights into the genetic features underlying the selection of efficient oxygen reducing microbes on biocathodes.

Anaerobic digestion of biowaste under extreme ammonia concentration: Identification of key microbial phylotypes.

Ammonia inhibition represents a major operational issue for anaerobic digestion (AD). In order to get more insights into AD microbiota resistance, anaerobic batch reactors performances were investigated under a wide range of Total Ammonia Nitrogen (TAN) concentrations up to 50.0g/L at 35°C. The half maximal inhibitory concentration (IC50) value was determined to be 19.0g/L. Microbial community dynamics revealed that above a TAN concentration of 10.0g/L, remarkable modifications within archaeal and bacterial communities occurred. 16S rRNA gene sequencing analysis showed a gradual methanogenic shift between two OTUs from genus Methanosarcina when TAN concentration increased up to 25.0g/L. Proportion of potential syntrophic microorganisms such as Methanoculleus and Treponema progressively raised with increasing TAN up to 10.0 and 25.0g/L respectively, while Syntrophomonas and Ruminococcus groups declined. In 25.0g/L assays, Caldicoprobacter were dominant. This study highlights the emergence of AD key phylotypes at extreme ammonia concentrations.

Increasing concentrations of phenol progressively affect anaerobic digestion of cellulose and associated microbial communities.

Performance stability is a key issue when managing anaerobic digesters. However it can be affected by external disturbances caused by micropollutants. In this study the influence of phenol on the methanization of cellulose was evaluated through batch toxicity assays. Special attention was given to the dynamics of microbial communities by means of automated ribosomal intergenic spacer analysis. We observed that, as phenol concentrations increased, the different steps of anaerobic cellulose digestion were unevenly and progressively affected, methanogenesis being the most sensitive: specific methanogenic activity was half-inhibited at 1.40 g/L of phenol, whereas hydrolysis of cellulose and its fermentation to VFA were observed at up to 2.00 g/L. Depending on the level of phenol, microbial communities resisted either through physiological or structural adaptation. Thus, performances at 0.50 g/L were maintained in spite of the microbial community's shift. However, the communities' ability to adapt was limited and performances decreased drastically beyond 2.00 g/L of phenol.

Asymmetrical response of anaerobic digestion microbiota to temperature changes.

In natural settings, anaerobic digestion can take place in a wide temperature range, but industrial digesters are usually operated under either mesophilic (~35 °C) or thermophilic (~55 °C) conditions. The ability of anaerobic digestion microbiota to switch from one operating temperature to the other remains poorly documented. We therefore studied the effect of sudden temperature changes (35 °C/55 °C) in lab-scale bioreactors degrading 13C-labelled cellulose. An asymmetric behaviour was observed. In terms of methane production, after an adaptation period, mesophilic inoculum exhibited a functional resistance to temperature increase but no functional resilience when temperature was reset to 35 °C, while thermophilic inoculum methanogenic activity strongly decreased under mesophilic conditions but partially recovered when temperature was reset to 55 °C. Automated ribosomal intergenic spacer analysis community fingerprints evidenced a strong influence of temperature on microbial diversity, particularly pronounced and persistent for Archaea. Key phylotypes involved in 13C-cellulose degradation were identified with a coupled stable isotope probing (SIP)-16S rDNA pyrotag sequencing approach, suggesting that the hydrolytic and fermentative metabolic functions could be maintained thanks to functional redundancy between members of the class Clostridia, whereas methanogenic activity primarily relied on specialized groups affiliated either to genus Methanosarcina (mesophilic conditions), Methanothermobacter or Methanoculleus (thermophilic conditions) that were irreversibly modified by temperature increase.