Bruce E. Rittmann

Human gut microbiota in obesity and after gastric bypass

Recent evidence suggests that the microbial community in the human intestine may play an important role in the pathogenesis of obesity. We examined 184,094 sequences of microbial 16S rRNA genes from PCR amplicons by using the 454 pyrosequencing technology to compare the microbial community structures of 9 individuals, 3 in each of the categories of normal weight, morbidly obese, and post-gastric-bypass surgery. Phylogenetic analysis demonstrated that although the Bacteria in the human intestinal community were highly diverse, they fell mainly into 6 bacterial divisions that had distinct differences in the 3 study groups. Specifically, Firmicutes were dominant in normal-weight and obese individuals but significantly decreased in post-gastric-bypass individuals, who had a proportional increase of Gammaproteobacteria. Numbers of the H2-producing Prevotellaceae were highly enriched in the obese individuals. Unlike the highly diverse Bacteria, the Archaea comprised mainly members of the order Methanobacteriales, which are H2-oxidizing methanogens. Using real-time PCR, we detected significantly higher numbers of H2-utilizing methanogenic Archaea in obese individuals than in normal-weight or post-gastric-bypass individuals. The coexistence of H2-producing bacteria with relatively high numbers of H2-utilizing methanogenic Archaea in the gastrointestinal tract of obese individuals leads to the hypothesis that interspecies H2 transfer between bacterial and archaeal species is an important mechanism for increasing energy uptake by the human large intestine in obese persons. The large bacterial population shift seen in the post-gastric-bypass individuals may reflect the double impact of the gut alteration caused by the surgical procedure and the consequent changes in food ingestion and digestion.

Opportunities for renewable bioenergy using microorganisms

Global warming can be slowed, and perhaps reversed, only when society replaces fossil fuels with renewable, carbon-neutral alternatives. The best option is bioenergy: the suns energy is captured in biomass and converted to energy forms useful to modern society. To make a dent in global warming, bioenergy must be generated at a very high rate, since the world today uses approximately 10 TW of fossil-fuel energy. And, it must do so without inflicting serious damage on the environment or disrupting our food supply. While most bioenergy options fail on both counts, several microorganism-based options have the potential to produce large amounts of renewable energy without disruptions. In one approach, microbial communities convert the energy value of various biomass residuals to socially useful energy. Biomass residuals come from agricultural, animal, and a variety of industrial operations, as well as from human wastes. Microorganisms can convert almost all of the energy in these wastes to methane, hydrogen, and electricity. In a second approach, photosynthetic microorganisms convert sunlight into biodiesel. Certain algae (eukaryotes) or cyanobacteria (prokaryotes) have high lipid contents. Under proper conditions, these photosynthetic microorganisms can produce lipids for biodiesel with yields per unit area 100 times or more than possible with any plant system. In addition, the non-lipid biomass can be converted to methane, hydrogen, or electricity. Photosynthetic microorganisms do not require arable land, an advantage because our arable land must be used to produce food. Algae or cyanobacteria may be the best option to produce bioenergy at rates high enough to replace a substantial fraction of our societys use of fossil fuels.

Gut Microbiota and Its Possible Relationship With Obesity

Obesity results from alterations in the bodys regulation of energy intake, expenditure, and storage. Recent evidence, primarily from investigations in animal models, suggests that the gut microbiota affects nutrient acquisition and energy regulation. Its composition has also been shown to differ in lean vs obese animals and humans. In this article, we review the published evidence supporting the potential role of the gut microbiota in the development of obesity and explore the role that modifying the gut microbiota may play in its future treatment. Evidence suggests that the metabolic activities of the gut microbiota facilitate the extraction of calories from ingested dietary substances and help to store these calories in host adipose tissue for later use. Furthermore, the gut bacterial flora of obese mice and humans include fewer Bacteroidetes and correspondingly more Firmicutes than that of their lean counterparts, suggesting that differences in caloric extraction of ingested food substances may be due to the composition of the gut microbiota. Bacterial lipopolysaccharide derived from the intestinal microbiota may act as a triggering factor linking inflammation to high-fat diet-induced metabolic syndrome. Interactions among microorganisms in the gut appear to have an important role in host energy homeostasis, with hydrogen-oxidizing methanogens enhancing the metabolism of fermentative bacteria. Existing evidence warrants further investigation of the microbial ecology of the human gut and points to modification of the gut microbiota as one means to treat people who are over-weight or obese.

A kinetic perspective on extracellular electron transfer by anode-respiring bacteria.

In microbial fuel cells and electrolysis cells (MXCs), anode-respiring bacteria (ARB) oxidize organic substrates to produce electrical current. In order to develop an electrical current, ARB must transfer electrons to a solid anode through extracellular electron transfer (EET). ARB use various EET mechanisms to transfer electrons to the anode, including direct contact through outer-membrane proteins, diffusion of soluble electron shuttles, and electron transport through solid components of the extracellular biofilm matrix. In this review, we perform a novel kinetic analysis of each EET mechanism by analyzing the results available in the literature. Our goal is to evaluate how well each EET mechanism can produce a high current density (> 10 A m(-2)) without a large anode potential loss (less than a few hundred millivolts), which are feasibility goals of MXCs. Direct contact of ARB to the anode cannot achieve high current densities due to the limited number of cells that can come in direct contact with the anode. Slow diffusive flux of electron shuttles at commonly observed concentrations limits current generation and results in high potential losses, as has been observed experimentally. Only electron transport through a solid conductive matrix can explain observations of high current densities and low anode potential losses. Thus, a study of the biological components that create a solid conductive matrix is of critical importance for understanding the function of ARB.

Proton transport inside the biofilm limits electrical current generation by anode‐respiring bacteria

Anode‐respiring bacteria (ARB) in a biofilm anode carry out an oxidation half‐reaction of organic matter, producing an electrical current from renewable biomass, including wastes. At the same time, ARB produce protons, usually one proton for every electron. Our study shows how current density generated by an acclimated ARB biofilm was limited by proton transport out of the biofilm. We determined that, at high current densities, protons were mainly transported out of the biofilm by protonating the conjugate base of the buffer system; the maximum current generation was directly related to the transport of the buffer, mainly by diffusion, into and out of the biofilm. With non‐limiting acetate concentrations, the current density increased with higher buffer concentrations, going from 2.21 ± 0.02 A m−2 with 12.5‐mM phosphate buffer medium to 9.3 ± 0.4 A m−2 using a 100‐mM phosphate buffer at a constant anode potential of Eanode = −0.35 V versus Ag/AgCl. Increasing the concentration of sodium chloride in the medium (0–100 mM) increased current density by only 15%, indicating that ion migration was not as important as diffusion of phosphate inside the biofilm. The current density also varied strongly with medium pH as a result of the buffer speciation: The current density was 10.0 ± 0.8 A m−2 at pH 8, and the pH giving one‐half the maximum rate was 6.5. A j–V curve analysis using 100 mM phosphate buffer showed a maximum current density of 11.5 ± 0.9 A m−2 and half‐saturation potential of −0.414 V versus Ag/AgCl, a value that deviated only slightly from the standard acetate potential, resulting in small anode‐potential losses. We discuss the implications of the proton‐transport limitation in the field of microbial fuel cells and microbial electrolytic cells. Biotechnol. Bioeng. 2008;100: 872–881.

Selecting Anode-Respiring Bacteria Based on Anode Potential: Phylogenetic, Electrochemical, and Microscopic Characterization

Anode-respiring bacteria (ARB) are able to transfer electrons contained in organic substrates to a solid electrode. The selection of ARB should depend on the anode potential, which determines the amount of energy available for bacterial growth and maintenance. In our study, we investigated how anode potential affected the microbial diversity of the biofilm community. We used a microbial electrolysis cell (MEC) containing four graphite electrodes, each at a different anode potential (E(anode) = -0.15, -0.09, +0.02, and +0.37 V vs SHE). We used wastewater-activated sludge as inoculum, acetate as substrate, and continuous-flow operation. The two electrodes at the lowest potentials showed a faster biofilm growth and produced the highest current densities, reaching up to 10.3 A/m(2) at the saturation of an amperometric curve; the electrode at the highest potential produced a maximum of 0.6 A/m(2). At low anode potentials, clone libraries showed a strong selection (92-99% of total clones) of an ARB that is 97% similar to G. sulfurreducens. At the highest anode potential, the ARB community was diverse. Cyclic voltammograms performed on each electrode suggest that the ARB grown at the lowest potentials carried out extracellular electron transport exclusively by conducting electrons through the extracellular biofilm matrix. This is supported by scanning electron micrographs showing putative bacterial nanowires and copious EPS at the lowest potentials. Non-ARB and ARB using electron shuttles in the diverse community for the highest anode potential may have insulated the ARB using a solid conductive matrix from the anode. Continuous-flow operation and the selective pressure due to low anode potentials selected for G. sulfurreducens, which are known to consume acetate efficiently and use a solid conductive matrix for electron transport.

Biological hydrogen production: prospects and challenges

Hydrogen gas provides exceptional value as an energy carrier and industrial feedstock, but currently is produced entirely by reforming fossil fuels. Biological hydrogen production (BioH(2)), which offers the possibility of being renewable and carbon neutral, can be achieved by photosynthesis, fermentation, and microbial electrolysis cells. This review introduces the principles, advantages and challenges of each approach to BioH(2). Photosynthetic BioH(2) is the ultimate renewable source, since it directly uses inexhaustible resources: sunlight energy and electrons from H(2)O. However, it presents major technical challenges, particularly due to oxygen sensitivity. Fermentative BioH(2) offers a high production rate, but poor conversion efficiency from the organic substrate to H(2). The microbial electrolysis cell can achieve high conversion efficiency, but is an emerging technology.

Applying a novel autohydrogenotrophic hollow-fiber membrane biofilm reactor for denitrification of drinking water

We conducted a series of pseudo-steady-state experiments on a novel hollow-fiber membrane biofilm reactor used for denitrification of oligotrophic waters, such as drinking water. We applied a range of nitrate loadings and hydrogen pressures to establish under what conditions the system could attain three goodness-of-performance criteria: partial nitrate removal, minimization of hydrogen wasting, and low nitrite accumulation. The hollow-fiber membrane biofilm reactor could meet drinking-water standards for nitrate and nitrite while minimizing the amount of hydrogen wasted in the effluent when it was operated under hydrogen-limited conditions. For example, the system could achieve partial nitrate removals between 39% and 92%, effluent nitrate between 0.4 and 9.1 mg N/l, effluent nitrite less than 1 mg N/l, and effluent hydrogen below 0.1 mg H2/l. High fluxes of nitrate and hydrogen made it possible to have a short liquid retention time (42 min), compared with 1-13 h in other studies with hydrogen used as the electron donor for denitrification. The fluxes and concentrations for hydrogen, nitrate, and nitrite obtained in this study can be used as practical guidelines for system design.

Occurrence of disinfection byproducts in United States wastewater treatment plant effluents

Effluents from wastewater treatment plants (WWTPs) contain disinfection byproducts (DBPs) of health concern when the water is utilized downstream as a potable water supply. The pattern of DBP formation was strongly affected by whether or not the WWTP achieved good nitrification. Chlorine addition to poorly nitrified effluents formed low levels of halogenated DBPs, except for (in some cases) dihalogenated acetic acids, but often substantial amounts of N-nitrosodimethyamine (NDMA). Chlorination of well-nitrified effluent typically resulted in substantial formation of halogenated DBPs but much less NDMA. For example, on a median basis after chlorine addition, the well-nitrified effluents had 57 microg/L of trihalomethanes [THMs] and 3 ng/L of NDMA, while the poorly nitrified effluents had 2 microg/L of THMs and 11 ng/L of NDMA. DBPs with amino acid precursors (haloacetonitriles, haloacetaldehydes) formed at substantial levels after chlorination of well-nitrified effluent. The formation of halogenated DBPs but not that of NDMA correlated with the formation of THMs in WWTP effluents disinfected with free chlorine. However, THM formation did not correlate with the formation of other DBPs in effluents disinfected with chloramines. Because of the relatively high levels of bromide in treated wastewater, bromine incorporation was observed in various classes of DBPs.

Non-steady state modeling of extracellular polymeric substances, soluble microbial products, and active and inert biomass

We present a modeling approach that quantifies the unified theory presented in the companion paper. In this approach, we use mathematical modeling to quantify the relationships among three solid species--bacteria, extracellular polymeric substances (EPS), and inert residual biomass-two soluble microbial products (SMP), original substrate, and an electron acceptor. According to the model, donor electrons are used for the synthesis of biomass, EPS, and utilization-associated products. Residual inert biomass and biomass-associated products are produced from the decay of active biomass and the hydrolysis of EPS, respectively. The model includes mass balance equations that consistently describe the flow of electrons among the components. It is solved with a set of parameters appropriate to the experimental study of Hsieh et al. (Biotech. Bioeng. 44 (1994) 219). Model outputs capture all trends observed in steady-state CSTR experiments and transient batch experiments. This agreement supports that the unified theory correctly captures the interconnections among SMP, EPS, and active and inert biomass.