Largus T. Angenent

Succession of microbial consortia in the developing infant gut microbiome

The colonization process of the infant gut microbiome has been called chaotic, but this view could reflect insufficient documentation of the factors affecting the microbiome. We performed a 2.5-y case study of the assembly of the human infant gut microbiome, to relate life events to microbiome composition and function. Sixty fecal samples were collected from a healthy infant along with a diary of diet and health status. Analysis of >300,000 16S rRNA genes indicated that the phylogenetic diversity of the microbiome increased gradually over time and that changes in community composition conformed to a smooth temporal gradient. In contrast, major taxonomic groups showed abrupt shifts in abundance corresponding to changes in diet or health. Community assembly was nonrandom: we observed discrete steps of bacterial succession punctuated by life events. Furthermore, analysis of ≈500,000 DNA metagenomic reads from 12 fecal samples revealed that the earliest microbiome was enriched in genes facilitating lactate utilization, and that functional genes involved in plant polysaccharide metabolism were present before the introduction of solid food, priming the infant gut for an adult diet. However, ingestion of table foods caused a sustained increase in the abundance of Bacteroidetes, elevated fecal short chain fatty acid levels, enrichment of genes associated with carbohydrate utilization, vitamin biosynthesis, and xenobiotic degradation, and a more stable community composition, all of which are characteristic of the adult microbiome. This study revealed that seemingly chaotic shifts in the microbiome are associated with life events; however, additional experiments ought to be conducted to assess how different infants respond to similar life events.

Host remodeling of the gut microbiome and metabolic changes during pregnancy.

Many of the immune and metabolic changes occurring during normal pregnancy also describe metabolic syndrome. Gut microbiota can cause symptoms of metabolic syndrome in nonpregnant hosts. Here, to explore their role in pregnancy, we characterized fecal bacteria of 91 pregnant women of varying prepregnancy BMIs and gestational diabetes status and their infants. Similarities between infant-mother microbiotas increased with childrens age, and the infant microbiota was unaffected by mothers health status. Gut microbiota changed dramatically from first (T1) to third (T3) trimesters, with vast expansion of diversity between mothers, an overall increase in Proteobacteria and Actinobacteria, and reduced richness. T3 stool showed strongest signs of inflammation and energy loss; however, microbiome gene repertoires were constant between trimesters. When transferred to germ-free mice, T3 microbiota induced greater adiposity and insulin insensitivity compared to T1. Our findings indicate that host-microbial interactions that impact host metabolism can occur and may be beneficial in pregnancy.

Cathodes as electron donors for microbial metabolism: Which extracellular electron transfer mechanisms are involved?

This review illuminates extracellular electron transfer mechanisms that may be involved in microbial bioelectrochemical systems with biocathodes. Microbially-catalyzed cathodes are evolving for new bioprocessing applications for waste(water) treatment, carbon dioxide fixation, chemical product formation, or bioremediation. Extracellular electron transfer processes in biological anodes, were the electrode serves as electron acceptor, have been widely studied. However, for biological cathodes the question remains: what are the biochemical mechanisms for the extracellular electron transfer from a cathode (electron donor) to a microorganism? This question was approached by not only analysing the literature on biocathodes, but also by investigating known extracellular microbial oxidation reactions in environmental processes. Here, it is predicted that in direct electron transfer reactions, c-type cytochromes often together with hydrogenases play a critical role and that, in mediated electron transfer reactions, natural redox mediators, such as PQQ, will be involved in the bioelectrochemical reaction. These mechanisms are very similar to processes at the bioanode, but the components operate at different redox potentials. The biocatalyzed cathode reactions, thereby, are not necessarily energy conserving for the microorganism.

Bacterial community structures are unique and resilient in full-scale bioenergy systems

Anaerobic digestion is the most successful bioenergy technology worldwide with, at its core, undefined microbial communities that have poorly understood dynamics. Here, we investigated the relationships of bacterial community structure (>400,000 16S rRNA gene sequences for 112 samples) with function (i.e., bioreactor performance) and environment (i.e., operating conditions) in a yearlong monthly time series of nine full-scale bioreactor facilities treating brewery wastewater (>20,000 measurements). Each of the nine facilities had a unique community structure with an unprecedented level of stability. Using machine learning, we identified a small subset of operational taxonomic units (OTUs; 145 out of 4,962), which predicted the location of the facility of origin for almost every sample (96.4% accuracy). Of these 145 OTUs, syntrophic bacteria were systematically overrepresented, demonstrating that syntrophs rebounded following disturbances. This indicates that resilience, rather than dynamic competition, played an important role in maintaining the necessary syntrophic populations. In addition, we explained the observed phylogenetic differences between all samples on the basis of a subset of environmental gradients (using constrained ordination) and found stronger relationships between community structure and its function rather than its environment. These relationships were strongest for two performance variables—methanogenic activity and substrate removal efficiency—both of which were also affected by microbial ecology because these variables were correlated with community evenness (at any given time) and variability in phylogenetic structure (over time), respectively. Thus, we quantified relationships between community structure and function, which opens the door to engineer communities with superior functions.

Impact of training sets on classification of high-throughput bacterial 16s rRNA gene surveys.

Taxonomic classification of the thousands–millions of 16S rRNA gene sequences generated in microbiome studies is often achieved using a naïve Bayesian classifier (for example, the Ribosomal Database Project II (RDP) classifier), due to favorable trade-offs among automation, speed and accuracy. The resulting classification depends on the reference sequences and taxonomic hierarchy used to train the model; although the influence of primer sets and classification algorithms have been explored in detail, the influence of training set has not been characterized. We compared classification results obtained using three different publicly available databases as training sets, applied to five different bacterial 16S rRNA gene pyrosequencing data sets generated (from human body, mouse gut, python gut, soil and anaerobic digester samples). We observed numerous advantages to using the largest, most diverse training set available, that we constructed from the Greengenes (GG) bacterial/archaeal 16S rRNA gene sequence database and the latest GG taxonomy. Phylogenetic clusters of previously unclassified experimental sequences were identified with notable improvements (for example, 50% reduction in reads unclassified at the phylum level in mouse gut, soil and anaerobic digester samples), especially for phylotypes belonging to specific phyla (Tenericutes, Chloroflexi, Synergistetes and Candidate phyla TM6, TM7). Trimming the reference sequences to the primer region resulted in systematic improvements in classification depth, and greatest gains at higher confidence thresholds. Phylotypes unclassified at the genus level represented a greater proportion of the total community variation than classified operational taxonomic units in mouse gut and anaerobic digester samples, underscoring the need for greater diversity in existing reference databases.

Biochemical methane potential and biodegradability of complex organic substrates.

The biomethane potential and biodegradability of an array of substrates with highly heterogeneous characteristics, including mono- and co-digestion samples with dairy manure, was determined using the biochemical methane potential (BMP) assay. In addition, the ability of two theoretical methods to estimate the biomethane potential of substrates and the influence of biodegradability was evaluated. The results of about 175 individual BMP assays indicate that substrates rich in lipids and easily-degradable carbohydrates yield the highest methane potential, while more recalcitrant substrates with a high lignocellulosic fraction have the lowest. Co-digestion of dairy manure with easily-degradable substrates increases the specific methane yields when compared to manure-only digestion. Additionally, biomethane potential of some co-digestion mixtures suggested synergistic activity. Evaluated theoretical methods consistently over-estimated experimentally-obtained methane yields when substrate biodegradability was not accounted. Upon correcting the results of theoretical methods with observed biodegradability data, an agreement greater than 90% was achieved.

Light energy to bioelectricity: photosynthetic microbial fuel cells.

Here, we reviewed five different approaches that integrate photosynthesis with microbial fuel cells (MFCs)-photoMFCs. Until now, no conclusive report has been published that identifies direct electron transfer (DET) between a photosynthetic biocatalyst and the anode of a MFC. Therefore, most recent research has been performed to generate sufficient electric current from sunlight with either electrocatalysts or heterotrophic bacteria on the anode to convert photosynthetic products indirectly. The most promising photoMFCs to date are electrocatalytic bioelectrochemical systems (BESs) that convert hydrogen from photosynthesis and sediment-based BESs that can convert excreted organics from cyanobacteria or plants. In addition, illumination on the cathode may provide either oxygen for an electrocatalytic reduction reaction or a promising anoxygenic biocathode.

Bioelectrochemical systems: From extracellular electron transfer to biotechnological application

In the context of wastewater treatment, Bioelectrochemical Systems (BESs) have gained considerable interest in the past few years, and several BES processes are on the brink of application to this area. This book, written by a large number of world experts in the different sub-topics, describes the different aspects and processes relevant to their development. Bioelectrochemical Systems (BESs) use micro-organisms to catalyze an oxidation and/or reduction reaction at an anodic and cathodic electrode respectively. Briefly, at an anode oxidation of organic and inorganic electron donors can occur. Prime examples of such electron donors are waste organics and sulfides. At the cathode, an electron acceptor such as oxygen or nitrate can be reduced. The anode and the cathode are connected through an electrical circuit. If electrical power is harvested from this circuit, the system is called a Microbial Fuel Cell; if electrical power is invested, the system is called a Microbial Electrolysis Cell. The overall framework of bio-energy and bio-fuels is discussed. A number of chapters discuss the basics - microbiology, microbial ecology, electrochemistry, technology and materials development. The book continues by highlighting the plurality of processes based on BES technology already in existence, going from wastewater based reactors to sediment based bio-batteries. The integration of BESs into existing water or process lines is discussed. Finally, an outlook is provided of how BES will fit within the emerging biorefinery area. This title belongs to Integrated Environmental Technology Series . ISBN: 9781843392330 (Print) ISBN: 9781780401621 (eBook)

Microbial electrochemistry and technology: terminology and classification

Microbial electrochemistry is the study and application of interactions between living microbial cells and electrodes (i.e. electron conductors, capacitive materials). For a long time this subfield of bioelectrochemistry has been the interest of mainly fundamental researchers. This has considerably changed during the last decade and microbial electrochemistry gained interest from applied researchers and engineers. These researchers took the microbial fuel cell (MFC), which is a system that converts the chemical energy of organic material in wastewater into electric power, from a concept to a technology. In addition, a plethora of derivative technologies, such as microbial electrolysis cells (MECs), microbial desalination cells (MDCs), photomicrobial fuel cells (photoMFCs), microbial electrosynthesis (MES), and biocomputing have been developed. The growing number of systems is often referred to in literature under the termini bioelectrochemical system (BES), microbial electrochemical technology (MET), or electrobiotechnology. Within this article we introduce a classification of technologies based on interfacing microbiology and electrochemistry. We argue that BESs comprise all systems based on bioelectrochemistry, with a further layer of termini through the use of METs. Primary METs are based on extracellular electron transfer (direct or mediated), whereas secondary METs include systems in which electrochemistry is connected – at least through ionic contact – with a microbial process via the electrochemical control or adaptation of environmental parameters, such as pH or metabolite concentration level.

Innate and Adaptive Immunity Interact to Quench Microbiome Flagellar Motility in the Gut

Gut mucosal barrier breakdown and inflammation have been associated with high levels of flagellin, the principal bacterial flagellar protein. Although several gut commensals can produce flagella, flagellin levels are low in the healthy gut, suggesting the existence of control mechanisms. We find that mice lacking the flagellin receptor Toll-like receptor 5 (TLR5) exhibit a profound loss of flagellin-specific immunoglobulins (Igs) despite higher total Ig levels in the gut. Ribotyping of IgA-coated cecal microbiota showed Proteobacteria evading antibody coating in the TLR5(-/-) gut. A diversity of microbiome members overexpressed flagellar genes in the TLR5(-/-) host. Proteobacteria and Firmicutes penetrated small intestinal villi, and flagellated bacteria breached the colonic mucosal barrier. In vitro, flagellin-specific Ig inhibited bacterial motility and downregulated flagellar gene expression. Thus, innate-immunity-directed development of flagellin-specific adaptive immune responses can modulate the microbiomes production of flagella in a three-way interaction that helps to maintain mucosal barrier integrity and homeostasis.