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.

Factors shaping the abundance and diversity of the gut archaeome across the animal kingdom.

Archaea are common constituents of the gut microbiome of humans, ruminants, and termites but little is known about their diversity and abundance in other animals. Here, we analyse sequencing and quantification data of archaeal and bacterial 16S rRNA genes from 250 species of animals covering a large taxonomic spectrum. We detect the presence of archaea in 175 animal species belonging to invertebrates, fish, amphibians, birds, reptiles and mammals. We identify five dominant gut lineages, corresponding to Methanobrevibacter, Methanosphaera, Methanocorpusculum, Methanimicrococcus and "Ca. Methanomethylophilaceae". Some archaeal clades, notably within Methanobrevibacter, are associated to certain hosts, suggesting specific adaptations. The non-methanogenic lineage Nitrososphaeraceae (Thaumarchaeota) is frequently present in animal samples, although at low abundance, but may have also adapted to the gut environment. Host phylogeny, diet type, fibre content, and intestinal tract physiology are major drivers of the diversity and abundance of the archaeome in mammals. The overall abundance of archaea is more influenced by these factors than that of bacteria. Methanogens reducing methyl-compounds with H2 can represent an important fraction of the overall methanogens in many animals. Together with CO2-reducing methanogens, they are influenced by diet and composition of gut bacteria. Our results provide key elements toward our understanding of the ecology of archaea in the gut, an emerging and important field of investigation.

Mechanisms underlying Clostridium pasteurianum's metabolic shift when grown with Geobacter sulfurreducens.

Recently, a study showed that glycerol fermentation by Clostridium pasteurianum could be metabolically redirected when the electroactive bacterium Geobacter sulfurreducens was added in the culture. It was assumed that this metabolic shift of the fermentative species resulted from an interspecies electron transfer. The aim of this study was to find out the mechanisms used for this interaction and how they affect the metabolism of C. pasteurianum. To get insights into the mechanisms involved, several coculture setups and RNA sequencing with differential expression analysis were performed. As a result, a putative interaction model was proposed: G. sulfurreducens produces cobamide molecules that possibly modify C. pasteurianum metabolic pathway at the key enzyme glycerol dehydratase, and affect its vanadium nitrogenase expression. In addition, the results suggested that G. sulfurreducens' electrons could enter C. pasteurianum through its transmembrane flavin-bound polyferredoxin and cellular cytochrome b5-rubredoxin interplay, putatively reinforcing the metabolic shift. Unravelling the mechanisms behind the interaction between fermentative and electroactive bacteria helps to better understand the role of bacterial interactions in fermentation setups. KEY POINTS: • C. pasteurianum-G. sulfurreducens interaction inducing a metabolic shift is mediated • C. pasteurianum's metabolic shift in coculture might be induced by cobamides • Electrons possibly enter C. pasteurianum through a multiflavin polyferredoxin.

Microbial Interactions as Drivers of a Nitrification Process in a Chemostat.

This article deals with the inclusion of microbial ecology measurements such as abundances of operational taxonomic units in bioprocess modelling. The first part presents the mathematical analysis of a model that may be framed within the class of Lotka-Volterra models fitted to experimental data in a chemostat setting where a nitrification process was operated for over 500 days. The limitations and the insights of such an approach are discussed. In the second part, the use of an optimal tracking technique (developed within the framework of control theory) for the integration of data from genetic sequencing in chemostat models is presented. The optimal tracking revisits the data used in the aforementioned chemostat setting. The resulting model is an explanatory model, not a predictive one, it is able to reconstruct the different forms of nitrogen in the reactor by using the abundances of the operational taxonomic units, providing some insights into the growth rate of microbes in a complex community.

Modeling of interspecies electron transfer in anaerobic microbial communities.

Interspecies electron transfer (IET) is a key phenomenon in anaerobic ecosystems, which is traditionally modeled as hydrogen transfer. Recently discovered alternative mediated IET (MIET) or direct IET (DIET) offer exciting alternative mechanisms of microbial partnerships that could lead to new strategies for the improvement of biotechnologies. Here, we analyze mathematical modeling of DIET and MIET in anaerobic ecosystems. Bioenergetics approaches already enable the evaluation of different energy sharing scenarios between microorganisms and give interesting clues on redox mediators and on possible ways of driving microbial communities relying on IET. The modeling of DIET kinetics however is currently only in its infancy. Recent concepts introduced for the modeling of electroactive biofilms should be further exploited. Recent modeling examples confirms the potential of DIET to increase the IET rates compared to H2-MIET, but also point out the need for additional characterizations of biological components supporting IET to improve predictions.

Mapping the biological activities of filamentous oxygenic photogranules.

Oxygenic photogranules have been suggested as alternatives to activated sludge in wastewater treatment. Challenging for modeling photogranule-based processes is the heterogeneity of photogranule morphologies, resulting in different activities by photogranule type. The measurement of microscale-activities of filamentous photogranules is particularly difficult because of their labile interfaces. We present here an experimental and modeling approach to quantify phototrophic O2 production, heterotrophic O2 consumption, and O2 diffusion in filamentous photogranules. We used planar optodes for the acquisition of spatio-temporal oxygen distributions combined with two-dimensional mathematical modeling. Light penetration into the photogranule was the factor controlling photogranule activities. The spatial distribution of heterotrophs and phototrophs had less impact. The photosynthetic response of filaments to light was detectable within seconds, emphasizing the need to analyze dynamics of light exposure of individual photogranules in photobioreactors. Studying other recurring photogranule morphologies will eventually enable the description of photogranule-based processes as the interplay of interacting photogranule populations.

Physical assessments of termites (Termitidae) under 2.45 GHz microwave irradiation.

Demands for chemical-free treatments for controlling insect pests are increasing worldwide. One such treatment is microwave heating; however, two critical issues arise when using microwaves as a heat source: intensive labor and excessive energy-consumption. Optimization is thus required to reduce energy consumption while effectively killing insects. Currently, the lethal effect of microwaves on insects is considered to be due to the temperature of the irradiated materials. This study examines how the conditions of irradiation, such as resonance or traveling mode, changed the conversion of electromagnetic energy into heat when 2.45 GHz microwaves penetrated the body of the termite, C. formosanus. Our results indicated that it is possible to heat and kill termites with microwaves under resonance condition. Termites were however found to be very tolerant to microwave irradiation as the permittivity of the insect was low compared with other reported insects and plants. Electron spin resonance revealed that termites contained several paramagnetic substances in their bodies, such as Fe3+, Cu2+, Mn2+, and organic radicals. Interestingly, irradiation with traveling microwaves hardly produced heat, but increased the organic radicals in termite bodies indicating non-thermal effects of microwaves.

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.

Bioflocculation and settling studies of native wastewater filamentous cyanobacteria using different cultivation systems for a low-cost and easy to control harvesting process.

Bioflocculation phenomena for filamentous cyanobacteria were studied and analysed in two different cultivation systems (i.e. based on air-bubbling and on shaking) and for different initial biomass concentrations. Floc formation and biomass settling were monitored during batch cultivation tests according to an innovative protocol. Results showed that the two cultivation systems enhanced two different flocculation behaviours: air bubbling led to the formation of small and dense flocs, while the shaking table resulted in larger (14 mm2 vs 4 mm2) but mechanically weaker flocs. Floc analysis evidenced that the different mixing systems also affected the speciation of biomass. A mathematical model was developed to simulate and predict the settling performance during the bioflocculation process of filamentous cyanobacteria. Natural settling was examined at different phases of biomass growth. Optimal conditions were obtained at the end of the exponential growth phase, when 70% of the total cultivated biomass could be recovered.

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.

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.

Revealing extracellular electron transfer mediated parasitism: energetic considerations.

Extracellular electron transfer (EET) is a mechanism that allows energetic coupling between two microorganisms or between a microorganism and an electrode surface. EET is either supported by direct physical contacts or mediated by electron shuttles. So far, studies dealing with interspecies EET (so-called IET) have mainly focused on possible syntrophic interactions between microorganisms favoured by this mechanism. In this article, the case of fermentative bacteria receiving extracellular electrons while fermenting a substrate is considered. A thermodynamical analysis based on metabolic energy balances was applied to re-investigate experimental data from the literature. Results suggest that the observations of a decrease of cell biomass yields of fermentative electron-accepting species, as mostly reported, can be unravelled by EET energetics and correspond to parasitism in case of IET. As an illustration, the growth yield decrease of Propionibacterium freudenreichii (-14%) observed in electro-fermentation experiments was fully explained by EET energetics when electrons were used by this species at a potential of -0.12 ± 0.01 V vs SHE. Analysis of other cases showed that, in addition to EET energetics in Clostridium pasteurianum, biological regulations can also be involved in such biomass yield decrease (-33% to -38%). Interestingly, the diminution of bacterial biomass production is always concomitant with an increased production of reduced compounds making IET-mediated parasitism and electro-fermentation attractive ways to optimize carbon fluxes in fermentation processes.

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.

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.

Shotgun metaproteomic profiling of biomimetic anaerobic digestion processes treating sewage sludge.

Two parallel anaerobic digestion lines were designed to match a "bovid-like" digestive structure. Each of the lines consisted of two continuous stirred tank reactors placed in series and separated by an acidic treatment step. The first line was inoculated with industrial inocula whereas the second was seeded with cow digestive tract contents. After 3 months of continuous sewage sludge feeding, samples were recovered for shotgun metaproteomic and DNA-based analysis. Strikingly, protein-inferred and 16S ribosomal DNA tags based taxonomic community profiles were not consistent. PCA however revealed a similar clustering pattern of the samples, suggesting that reproducible methodological and/or biological factors underlie this observation. The performances of the two digestion lines did not differ significantly and the cow-derived inocula did not establish in the reactors. A low throughput metagenomic dataset (3.4 × 10(6) reads, 1.1 Gb) was also generated for one of the samples. It allowed a substantial increase of the analysis depth (11 vs. 4% of spectral identification rate for the combined samples). Surprisingly, a high proportion of proteins from members of the "Candidatus Competibacter" group, a key microbial player usually found in activated sludge plants, was retrieved in our anaerobic digester samples. Data are available via ProteomeXchange with identifier PXD002420 (http://proteomecentral.proteomexchange.org/dataset/PXD002420).

Multi-system Nernst-Michaelis-Menten model applied to bioanodes formed from sewage sludge.

Bioanodes were formed under constant polarization at -0.2 V/SCE from fermented sewage sludge. Current densities reached were 9.3±1.2 A m(-2) with the whole fermented sludge and 6.2±0.9 A m(-2) with the fermented sludge supernatant. The bioanode kinetics was analysed by differentiating among the contributions of the three redox systems identified by voltammetry. Each system ensured reversible Nernstian electron transfer but around a different central potential. The global overpotential required to reach the maximum current plateau was not imposed by slow electron transfer rates but was due to the potential range covered by the different redox systems. The microbial communities of the three bioanodes were analysed by 16S rRNA gene pyrosequencing. They showed a significant microbial diversity around a core of Desulfuromonadales, the proportion of which was correlated with the electrochemical performance of the bioanodes.

Stable isotope probing of acetate fed anaerobic batch incubations shows a partial resistance of acetoclastic methanogenesis catalyzed by Methanosarcina to sudden increase of ammonia level.

Ammonia inhibition represents a major operational issue for anaerobic digestion. In order to refine our understanding of the terminal catabolic steps in thermophilic anaerobic digestion under ammonia stress, we studied batch thermophilic acetate fed experiments at low (0.26 g L(-1)) and high (7.00 g L(-1)) Total Ammonia Nitrogen concentrations (TAN). Although methane production started immediately for all incubations and resulted in methane yields close to stoichiometric expectations, a 62-72% decrease of methanogenic rate was observed throughout the incubation at 7.00 g L(-1) of TAN compared to 0.26 g L(-1). Stable Isotope Probing analysis of active microbial communities in (13)C-acetate fed experiments coupled to automated ribosomal intergenic spacer analysis and 16S rDNA pyrotag sequencing confirmed that microbial communities were similar for both TAN conditions. At both TAN levels, the (13)C-labeled bacterial community was mainly affiliated to Clostridia-relatives, with OPB54 bacteria being the most abundant sequence in the heavy DNA 16S rDNA pyrotag library. Sequences closely related to Methanosarcina thermophila were also abundantly retrieved in the heavy DNA fractions, showing that this methanogen was still actively assimilating labeled carbon from acetate at free ammonia nitrogen concentrations up to 916 mg L(-1). Stable isotopic signature analysis of biogas, measured in unlabeled acetate fed experiments that were conducted in parallel, confirmed that acetoclastic methanogenic pathway was dominant at both ammonia concentrations. Our work demonstrates that, besides the syntrophic acetate oxidation pathway, acetoclastic methanogenesis catalyzed by Methanosarcina can also play a major role in methane production at high ammonia levels.

The current provided by oxygen-reducing microbial cathodes is related to the composition of their bacterial community.

Oxygen reducing biocathodes were formed from sludge under constant polarization at -0.2 and +0.4V/SCE. Under chronoamperometry at pH10.3 ± 0.3, current densities of 0.21 ± 0.03 and 0.12 ± 0.01 Am(-2) were displayed at -0.2V/SCE by the biocathodes formed at -0.2 and 0.4V/SCE, respectively. Voltammetry revealed similar general characteristics for all biocathodes and higher diffusion-limited current densities (0.84 ± 0.26 Am(-2)) than chronoamperometry. Up to 3.7 Am(-2) was reached under air bubbling. A theoretical model was proposed to show the consistency of the chronoamperometric and voltammetric data. The biocathodes formed at -0.2V/ECS that gave the highest electrochemical performance showed a homogeneous selection of Deinococcus-Thermus and Gemmatimonadetes, while the biocathodes formed at 0.4V/SCE were enriched in different bacteria. The biocathode that led to the worst electrochemical characteristics, while formed at -0.2V/SCE, showed the largest bacterial diversity. The biocathode performance was consequently related to the enrichment in specific microbial phyla. Moreover, the strong presence of bacteria parented to Deinococci may also have some interest in biotechnology.

A thermodynamic theory of microbial growth.

Our ability to model the growth of microbes only relies on empirical laws, fundamentally restricting our understanding and predictive capacity in many environmental systems. In particular, the link between energy balances and growth dynamics is still not understood. Here we demonstrate a microbial growth equation relying on an explicit theoretical ground sustained by Boltzmann statistics, thus establishing a relationship between microbial growth rate and available energy. The validity of our equation was then questioned by analyzing the microbial isotopic fractionation phenomenon, which can be viewed as a kinetic consequence of the differences in energy contents of isotopic isomers used for growth. We illustrate how the associated theoretical predictions are actually consistent with recent experimental evidences. Our work links microbial population dynamics to the thermodynamic driving forces of the ecosystem, which opens the door to many biotechnological and ecological developments.