Biomass acid pretreatment impacts on metabolic routes and bacterial composition of dark fermentation process.

Complex organic matter represents a suitable substrate to produce hydrogen through dark fermentation (DF) process. To increase H2 yields, pretreatment technology is often required. The main objective of the present work was to investigate thermo-acid pretreatment impact on sugar solubilization and biotic parameters of DF of sorghum or organic fraction of municipal solid waste (OFMSW). Biochemical hydrogen potential tests were carried out without inoculum using raw or thermo-acid pretreated substrates. Results showed an improvement in sugar solubilization after thermo-acid pretreatments. Pretreatments led to similar DF performances (H2 and total metabolite production) compared to raw biomasses. Nevertheless, they were responsible for bacterial shifts from Enterobacteriales towards Clostridiales and Bacillales as well as metabolic changes from acetate towards butyrate or ethanol. The metabolic changes were attributed to the biomass pretreatment impact on indigenous bacteria as no change in the metabolic profile was observed after performing thermo-acid pretreatments on irradiated OFMSW (inactivated indigenous bacteria and inoculum addition). Consequently, acid pretreatments were inefficient to improve DF performances but led to metabolic and bacterial community changes due to their impact on indigenous bacteria.

Improving the storage of cover crops by co-ensiling with different waste types: Effect on fermentation and effluent production.

Cover crops harvested at a low maturity stage generally have a high moisture content, which may generate energy losses during silage storage via effluent production and undesirable fermentations. This paper investigates the use of different waste types as absorbent co-substrates to be added before ensiling. The relation between the absorbent water holding capacity and silage effluent volume was first studied to find an effective parameter to prevent effluent production. Effluent retention was found to be proportional to the absorbent loading and water holding capacity (r2 = 0.98) and up to 90 % of effluent production was avoided when compared to control (295 l.t-1). The impact of different co-substrates (including bio-waste and manures) on overall ensiling performances was then investigated at an optimized absorbent loading. All co-substrates allowed a total effluent retention while a 76 l.t-1 effluent volume was reported for the control. The silage fermentation was modified or mostly unchanged depending on the co-substrate chemical and microbial properties and different metabolic pathways were observed (e.g. homolactic or butyric fermentation). In most conditions, the methane potential of the crop was efficiently preserved over a storage of 60 days. Co-ensiling was shown to be a relevant silage preparation method for biogas production.

Conditions for efficient alkaline storage of cover crops for biomethane production.

An innovative process aiming to combine storage and alkali pretreatment of cover crops was investigated using lime as a low cost and environmental friendly reactant. Different lime loadings and Total Solid concentrations (TS) allowed to highlight the abiotic mechanisms of deacetylation during the early stages of the process. Long-term storage experiments of rye and sunflower cover crops at 100 g.kgTS-1 lime loading allowed to evaluate the fermentation kinetics and to compare performances in dry and wet conditions to classical silage storage. The dry condition allowed an efficient alkaline storage and up to a 15.7% Biochemical Methane Potential (BMP) increase, while the wet condition underwent a succession of fermentations with a high butyric acid accumulation and H2 production, leading to a 13% BMP loss. Silage experiments allowed an efficient preservation of the BMP, with no significant variation over the 6-month storage duration.

Microbial community redundance in biomethanation systems lead to faster recovery of methane production rates after starvation.

The Power-to-Gas concept corresponds to the use of the electric energy surplus to produce H2 by water electrolysis, that can be further converted to methane by biomethanation. However, the fluctuant production of renewable energy sources can lead to discontinuous H2 injections into the reactors, that may interfere with the adaptation of the microbial community to high H2 partial pressures. In this study, the response of the microbial community to H2 and organic feed starvation was evaluated in in-situ and ex-situ biomethanation. The fed-batch reactors were fed with acetate or glucose and H2, and one or four weeks of starvation periods were investigated. Methane productivity was mostly affected by the four-week starvation period. However, both in-situ and ex-situ biomethanation reactors recovered their methane production rate after starvation within approximately one-week of normal operation, while the anaerobic digestion (AD) reactors did not recover their performances even after 3 weeks of normal operation. The recovery failure of the AD reactors was probably related to a slow growth of the syntrophic and methanogen microorganisms, that led to a VFA accumulation. On the contrary, the faster recovery of both biomethanation reactors was related to the replacement of Methanoculleus sp. by Methanobacterium sp., restoring the methane production in the in-situ and ex-situ biomethanation reactors. This study has shown that biomethanation processes can respond favourably to the intermittent H2 addition without compromising their CH4 production performance.

Role of indigenous bacteria in dark fermentation of organic substrates.

Hydrogen production by dark fermentation of complex organic substrates, such as biowaste, can naturally take place with indigenous bacteria or by adding an external microbial inoculum issued from various natural environments. This study aims to determine whether indigenous bacteria associated with thermal pretreatment could impact dark fermentation performances. Biochemical hydrogen potential tests were carried out on seven organic substrates. Results showed a strong influence of the indigenous bacteria which are as effective as thermally pretreated exogenous bacteria to produce H2 and metabolites. High abundance in Clostridiales and/or Enterobacteriales was associated with high H2 yield. This study shows that no inoculum nor pretreatment are required to achieve satisfactory dark fermentation performances from organic waste.

Conservation of acquired morphology and community structure in aged biofilms after facing environmental stress.

The influence of growth history on biofilm morphology and microbial community structure is poorly studied despite its important role for biofilm development. Here, biofilms were exposed to a change in hydrodynamic conditions at different growth stages and we observed how biofilm age affected the change in morphology and bacterial community structure. Biofilms were developed in two bubble column reactors, one operated under constant shear stress and one under variable shear stress. Biofilms were transferred from one reactor to the other at different stages in their development by withdrawing and inserting the support medium from one reactor to the other. The developments of morphology and microbial community structure were followed by image analysis and molecular tools. When transferred early in biofilm development, biofilms adapted to the new hydrodynamic conditions and adopted features of the biofilm already developed in the receiving reactor. Biofilms transferred at a late state of biofilm development continued their initial trajectories of morphology and community development even in a new environment. These biofilms did not immediately adapt to their new environment and kept features acquired during their early growth phase, a property we called memory effect.