One of the most important biotechnological challenges is to develop environment friendly technologies to produce new sources of energy. Microbial production of biohydrogen through dark fermentation, by conversion of residual biomass, is an attractive solution for short-term development of bioH2 producing processes. Efficient biohydrogen production relies on complex mixed communities working in tight interaction. Species composition and functional traits are of crucial importance to maintain the ecosystem service. The analysis of microbial community revealed a wide phylogenetic diversity that contributes in different-and still mostly unclear-ways to hydrogen production. Bridging this gap of knowledge between microbial ecology features and ecosystem functionality is essential to optimize the bioprocess and develop strategies toward a maximization of the efficiency and stability of substrate conversion. The aim of this review is to provide a comprehensive overview of the most up-to-date biodata available and discuss the main microbial community features of biohydrogen engineered ecosystems, with a special emphasis on the crucial role of interactions and the relationships between species composition and ecosystem service. The elucidation of intricate relationships between community structure and ecosystem function would make possible to drive ecosystems toward an improved functionality on the basis of microbial ecology principles.

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To provide new insight into the dark fermentation process, a multi-lateral study was performed to study the microbiology of 20 different lab-scale bioreactors operated in four different countries (Brazil, Chile, Mexico, and Uruguay). Samples (29) were collected from bioreactors with different configurations, operation conditions, and performances. The microbial communities were analyzed using 16S rRNA genes 454 pyrosequencing. The results showed notably uneven communities with a high predominance of a particular genus. The phylum Firmicutes predominated in most of the samples, but the phyla Thermotogae or Proteobacteria dominated in a few samples. Genera from three physiological groups were detected: high-yield hydrogen producers (Clostridium, Kosmotoga, Enterobacter), fermenters with low-hydrogen yield (mostly from Veillonelaceae), and competitors (Lactobacillus). Inocula, reactor configurations, and substrates influence the microbial communities. This is the first joint effort that evaluates hydrogen-producing reactors and operational conditions from different countries and contributes to understand the dark fermentation process.

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Hydrogen is a promising alternative of clean energy carrier which can be biologically produced from glycerol-rich waste an abundant and economic source of substrate. However, continuous hydrogen-producing systems still need to be improved and in particular by manipulating the only few available operating conditions. The aim of this study was to investigate the effect of the two main operational parameters, ie. pH and hydraulic retention time (HRT) on hydrogen yields and microbial community structures. For that, a continuous stirred tank reactor (CSTR) was first inoculated with an enriched mixed microflora and was then fed with glycerol. A strong influence of these two operational parameters was shown on hydrogen yields where the maximum yield (0.58 ± 0.13 mol H2 mol-1 glycerol) was achieved at pH 5.5-HRT 12 h and was 28.5 times higher than the minimum (0.02 ± 0.02 mol H2 mol-1 glycerol) obtained at pH 5.0-HRT 14 h. Changes in most dominant microbial populations were mainly influenced by the pH. Interestingly, HRT parameter related to changes in the metabolic patterns and influenced the composition of subdominant Version postprint