The hyperthermophilic anaerobic bacterium, Caldicellulosiruptor bescii, is effective in degrading and solubilizing lignocellulosic materials. Laboratory studies have characterized the chemistry of the process for crystalline cellulose and switchgrass, but the data are insufficient for engineering commercial plants to use C. bescii for pre-digestion of waste streams. The purpose of this study is three-fold: 1) to identify any potential toxicities in C. bescii pre-digestion and biogas production from several wastes; 2) to determine the potential enhancement of biogas production by anaerobic digestion of pre-digested dairy manure and waste activated sludge; and 3) to identify variables that must be quantified and controlled for engineering commercial, continuous-flow systems for waste disposal and biogas production incorporating C. bescii pre-digestion. Tests were run at lab-, bench- and pilot plant-scale with C.bescii pre-digestion and controls run at 75°C and pH 7-8 followed by mesophilic anaerobic digestion at 37-41°C. The lab- and bench-scale tests demonstrate that C. bescii is capable of growing on several organic wastes and pre-digestion with C. bescii increases conversion of waste into biogas, typically by a factor of 2 or more. Incorporation of C. bescii pre-digestion in an optimized commercial system is predicted to provide 75-85% volatile solids conversion to biogas with 75% methane when digesting dairy manure and sewage sludge. Achieving these results at a commercial scale requires further work to quantify C. bescii growth and enzyme production rates, as well as rates of base- and enzyme-catalyzed hydrolysis of the polymeric materials, e.g., lignocellulose, in the waste in order to optimize retention times.

This paper builds a quantitative thermodynamic model for the microbial hydrolysis process (MHP, which uses Caldicellulosiruptor bescii at 75°C for pre-digestion) for producing biogas from a 5-10% aqueous suspension of dairy manure (naturally buffered near pH 7.8 by ammonium bicarbonate) by anaerobic digestion with a mix of acetoclastic and syntrophic methanogenesis. Standard Gibbs energy changes were calculated for the major reactions in pre-digestion, for reactions producing H2, acetate, and CO2 in the digester, and for methanogenesis reactions in the digester. The available data limit the study to analyzing reactions in the digester to reactions of short-chain volatile fatty acids anions. Results are presented as curves of ΔrxnG (Gibbs energy change) vs. acetate concentration. The H2(aq) concentration must be above 1.2×10-9 M to get significant syntrophic methanogenesis, i.e., for ΔrxnG to be negative. The results show syntrophic methanogenesis of propionate, butyrate, and valerate slows as acetate concentration increases because hydrogen production also decreases, and consequently, biogas production from syntrophic methanogenesis slows as acetate increases. Bicarbonate also inhibits both acetoclastic and syntrophic methanogenesis but is necessary to prevent acidification (souring) of the digester. At identical steady-state conditions, acetoclastic methanogenesis runs about 1.4 times faster than syntrophic methanogenesis. Because syntrophic methanogenesis produces acetate catabolized by acetoclastic methanogens, both types of methanogens are necessary to maximize biogas production. The culture in the digester is predicted to evolve to optimize the ratio of acetoclastic methanogens to syntrophic methanogens, a condition signaled by a constant, low acetate concentration in the digester effluent. Obtaining volatile solids reduction as high as 75% with MHP requires a feedstock with less than 25% lignin and a culture of acetoclastic methanogens and syntrophic methanogens and their symbiotic bacteria.

Heat stress negatively affects several physiological and biochemical processes in grapevine plants. In this work, two new methods, calorespirometry, which has been used to determine temperature adaptation in plants, and near-infrared (NIR) spectroscopy, which has been used to determine several grapevine-related traits and to discriminate among varieties, were tested to evaluate grapevine response to high temperatures. ‘Touriga Nacional’ variety grapevines, inoculated or not with Rhizoglomus irregulare or Funneliformis mosseae, were used in this study. Calorespirometric parameters and NIR spectra, as well as other parameters commonly used to assess heat injury in plants, were measured before and after high temperature exposure. Growth rate and substrate carbon conversion efficiency, calculated from calorespirometric measurements, and stomatal conductance, were the most sensitive parameters for discriminating among high temperature responses of control and inoculated grapevines. The results revealed that, although this vine variety can adapt its physiology to temperatures up to 40 °C, inoculation with R. irregulare could additionally help to sustain its growth, especially after heat shocks. Therefore, the combination of calorespirometry together with gas exchange measurements is a promising strategy for screening grapevine heat tolerance under controlled conditions and has high potential to be implemented in initial phases of plant breeding programs.