Christian Abendroth

Eubacteria and archaea communities in seven mesophile anaerobic digester plants in Germany.

BackgroundOnly a fraction of the microbial species used for anaerobic digestion in biogas production plants are methanogenic archaea. We have analyzed the taxonomic profiles of eubacteria and archaea, a set of chemical key parameters, and biogas production in samples from nine production plants in seven facilities in Thuringia, Germany, including co-digesters, leach-bed, and sewage sludge treatment plants. Reactors were sampled twice, at a 1-week interval, and three biological replicates were taken in each case.ResultsA complex taxonomic composition was found for both eubacteria and archaea, both of which strongly correlated with digester type. Plant-degrading Firmicutes as well as Bacteroidetes dominated eubacteria profiles in high biogas-producing co-digesters; whereas Bacteroidetes and Spirochaetes were the major phyla in leach-bed and sewage sludge digesters. Methanoculleus was the dominant archaea genus in co-digesters, whereas Methanosarcina and Methanosaeta were the most abundant methanogens in leachate from leach-bed and sewage sludge digesters, respectively.ConclusionsThis is one of the most comprehensive characterizations of the microbial communities of biogas-producing facilities. Bacterial profiles exhibited very low variation within replicates, including those of semi-solid samples; and, in general, low variation in time. However, facility type correlated closely with the bacterial profile: each of the three reactor types exhibited a characteristic eubacteria and archaea profile. Digesters operated with solid feedstock, and high biogas production correlated with abundance of plant degraders (Firmicutes) and biofilm-forming methanogens (Methanoculleus spp.). By contrast, low biogas-producing sewage sludge treatment digesters correlated with high titers of volatile fatty acid-adapted Methanosaeta spp.

Producing high-strength liquor from mesophilic batch acidification of chicken manure

This report describes the results from anaerobic batch acidification of chicken manure as a mono-substrate studied under mesophilic conditions. The manure was diluted with tap water to prevent methane formation during acidification and to improve mixing conditions by reducing fluid viscosity; no anaerobic digester sludge has been added as an inoculum. Highest acidification rates were measured at concentrations of 10 gVS L−1 and 20 gVS L−1; the pH value remained high (pH 6.9–7.9) throughout the test duration and unexpected fast methane formation was observed in every single batch. At substrate concentrations of 10 gVS L−1 there was a remarkable methane formation representing a value of 82% of the respective biochemical methane potential of chicken manure. Increasing substrate concentrations did not supress methane formation but impaired acid production. Consequently, the liquor cannot be stored over longer periods but should immediately be used in a digestion process.

The CENP-T C-Terminus Is Exclusively Proximal to H3.1 and not to H3.2 or H3.3

The kinetochore proteins assemble onto centromeric chromatin and regulate DNA segregation during cell division. The inner kinetochore proteins bind centromeres while most outer kinetochore proteins assemble at centromeres during mitosis, connecting the complex to microtubules. The centromere–kinetochore complex contains specific nucleosomes and nucleosomal particles. CENP-A replaces canonical H3 in centromeric nucleosomes, defining centromeric chromatin. Next to CENP-A, the CCAN multi-protein complex settles which contains CENP-T/W/S/X. These four proteins are described to form a nucleosomal particle at centromeres. We had found the CENP-T C-terminus and the CENP-S termini next to histone H3.1 but not to CENP-A, suggesting that the Constitutive Centromere-Associated Network (CCAN) bridges a CENP-A- and a H3-containing nucleosome. Here, we show by in vivo FRET that this proximity between CENP-T and H3 is specific for H3.1 but neither for the H3.1 mutants H3.1C96A and H3.1C110A nor for H3.2 or H3.3. We also found CENP-M next to H3.1 but not to these H3.1 mutants. Consistently, we detected CENP-M next to CENP-S. These data elucidate the local molecular neighborhood of CCAN proteins next to a H3.1-containing centromeric nucleosome. They also indicate an exclusive position of H3.1 clearly distinct from H3.2, thus documenting a local, and potentially also functional, difference between H3.1 and H3.2.

Towards a Microbial Thermoelectric Cell

Microbial growth is an exothermic process. Biotechnological industries produce large amounts of heat, usually considered an undesirable by-product. In this work, we report the construction and characterization of the first microbial thermoelectric cell (MTC), in which the metabolic heat produced by a thermally insulated microbial culture is partially converted into electricity through a thermoelectric device optimized for low ΔT values. A temperature of 41°C and net electric voltage of around 250–600 mV was achieved with 1.7 L baker’s yeast culture. This is the first time microbial metabolic energy has been converted into electricity with an ad hoc thermoelectric device. These results might contribute towards developing a novel strategy to harvest excess heat in the biotechnology industry, in processes such as ethanol fermentation, auto thermal aerobic digestion (ATAD) or bioremediation, which could be coupled with MTCs in a single unit to produce electricity as a valuable by-product of the primary biotechnological product. Additionally, we propose that small portable MTCs could be conceived and inoculated with suitable thermophilic of hyperthermophilic starter cultures and used for powering small electric devices.

Microbial communities involved in biogas production exhibit high resilience to heat shocks

We report here the impact of heat-shock treatments (55 and 70 °C) on the biogas production within the acidification stage of a two-stage reactor system for anaerobic digestion and biomethanation of grass. The microbiome proved both taxonomically and functionally very robust, since heat shocks caused minor community shifts compared to the controls, and biogas yield was not decreased. The strongest impact on the microbial profile was observed with a combination of heat shock and low pH. Since no transient reduction of microbial diversity occured after the shock, biogas keyplayers, but also potential pathogens, survived the treatment. All along the experiment, the heat-resistant bacterial profile consisted mainly of Firmicutes, Bacteroidetes and Proteobacteria. Bacteroides and Acholeplasma were reduced after heat shocks. An increase was observed for Aminobacterium. Our results prove the stability to thermal stresses of the microbial communities involved in acidification, and the resilience in biogas production irrespectively of the thermal treatment.

Potential pitfalls of FISH microscopy as assessment method for anaerobic digesters

In the present work we investigated how the state of a biogas reactor impacts the enumeration of prokaryotic cells by fluorescence in situ hybridisation (FISH). Therefore, the correlation between gas production and FISH hybridisation rates was analysed in different anaerobic digester sludges. High gasification activity coincided with high hybridisation rates. Low hybridisation rates were especially achieved with reactor samples subjected to long starvation periods showing low biogas production. Based on our findings we conclude that samples for FISH analysis should be fixed as soon as possible to prevent a loss of microbial activity resulting in lower FISH signals. Furthermore, the location of sampling is of importance, since samples from different fermenters within the same biogas plant also varied strongly in their FISH hybridisation rate. Our results indicate that FISH could be a useful method for assessing the metabolic state of microorganisms in anaerobic digester plants.

Liquid co-substrates repower sewage microbiomes

A range of parameters are known to shape the methanogenic communities of biogas-producing digesters and to strongly influence the amount of biogas produced. In this work, liquid and solid fractions of grass biomass were used separately for semicontinuous batch methanation using sewage sludge as seed sludge. During 6 months of incubation, the amount of input COD was increased gradually, and the underlying methanogenic microbiome was assessed by means of microscopy-based automated cell counting and full-length 16S rRNA high-throughput sequencing. In this sense, we prove for the first time the suitability of the ONT™MinION platform as a monitoring tool for anaerobic digestion systems. According to our results, solid-fed batches were highly unstable at higher COD input concentrations, and kept Methanosaeta spp. typically associated to sewage sludge-as the majoritary methanogenic archaea. In contrast, liquid-fed batches developed a more stable microbiome, proved enriched in Methanosarcina spp, and resulted in higher methanogenic yield. This work demonstrates the high repowering potential of microbiomes from sewage sludge digesters, and highlight the effectiveness of liquefied substrates for increasing biogas productivity in anaerobic digestions.

Methanogenic community shifts during the transition from sewage mono-digestion to co-digestion of grass biomass

In this work, liquid and solid fractions of grass biomass were used as co-substrates for anaerobic co-digestion of sewage sludge. The input of grass biomass was increased gradually, and the underlying methanogenic microbiome was assessed by means of microscopy-based cell counting and full-length 16S rRNA gene high-throughput sequencing, proving for the first time the suitability of nanopore-based portable sequencers as a monitoring tool for anaerobic digestion systems. In both cases co-fermentation resulted in an increased number of bacteria and methanogenic archaea. Interestingly, the microbial communities were highly different between solid and liquid-fed batches. Liquid-fed batches developed a more stable microbiome, enriched in Methanosarcina spp., and resulted in higher methanogenic yield. In contrast, solid-fed batches were highly unstable at higher substrate concentrations, and kept Methanosaeta spp. - typically associated to sewage sludge - as the majoritary methanogenic archaea.

Warm and wet: robust lipase-producing bacteria from the indoor environment

Lipases are key biocatalysts with important biotechnological applications. With the aim of isolating robust lipolytic microbial strains, we have analyzed the bacterial communities inhabiting two domestic extreme environments: a thermophilic sauna and a dishwasher filter. Scanning electron microscopy revealed biofilm-forming and scattered microorganisms in the sauna and dishwasher sample, respectively. A culture-independent approach based on 16S rRNA analysis indicated a high abundance of Proteobacteria in the sauna sample; and, a large amount of Proteobacteria, Firmicutes, Cyanobacteria and Actinobacteria in the dishwasher filter. With a culture-dependent approach, we isolated 48 bacterial strains, screened their lipolytic activities on media with tributyrin as the main carbon source, and finally selected five isolates for further characterization. These strains, all of them identified as members of the genus Bacillus, displayed optimum lipolytic peaks at pH 6.5 and with 1-2% NaCl, and the activity proved very robust at a wide range of pH (up to 11.5) and added NaCl concentrations (up to 4%). The thermal, pH and salt robustness of the selected isolates is a valuable attribute for these strains, which are promising as highly tolerant biodetergents. To our knowledge, this is the first report regarding the isolation from an indoor environment of Bacillus strains with a high potential for industry.