Davide De Francisci

Microbial diversity and dynamicity of biogas reactors due to radical changes of feedstock composition

The anaerobic digestion process is often inhibited by alteration of substrates and/or organic overload. This study aimed to elucidate changes of microbial ecology in biogas reactors upon radical changes of substrates and to determine their importance to process imbalance. For this reason, continuously fed reactors were disturbed with pulses of proteins, lipids and carbohydrates and the microbial ecology of the reactors were characterized by 16S rRNA gene sequencing before and after the imposed changes. The microbial composition of the three reactors, initially similar, diverged greatly after substrate change. The greatest increase in diversity was observed in the reactor supplemented with carbohydrates and the microbial community became dominated by lactobacilli, while the lowest corresponded to the reactor overfed with proteins, where only Desulfotomaculum showed significant increase. The overall results suggest that feed composition has a decisive impact on the microbial composition of the reactors, and thereby on their performance.

New steady-state microbial community compositions and process performances in biogas reactors induced by temperature disturbances

BackgroundThe microbial community in a biogas reactor greatly influences the process performance. However, only the effects of deterministic factors (such as temperature and hydraulic retention time (HRT)) on the microbial community and performance have been investigated in biogas reactors. Little is known about the manner in which stochastic factors (for example, stochastic birth, death, colonization, and extinction) and disturbance affect the stable-state microbial community and reactor performances.ResultsIn the present study, three replicate biogas reactors treating cattle manure were run to examine the role of stochastic factors and disturbance in shaping microbial communities. In the triplicate biogas reactors with the same inoculum and operational conditions, similar process performances and microbial community profiles were observed under steady-state conditions. This indicated that stochastic factors had a minor role in shaping the profile of the microbial community composition and activity in biogas reactors. On the contrary, temperature disturbance was found to play an important role in the microbial community composition as well as process performance for biogas reactors. Although three different temperature disturbances were applied to each biogas reactor, the increased methane yields (around 10% higher) and decreased volatile fatty acids (VFAs) concentrations at steady state were found in all three reactors after the temperature disturbances. After the temperature disturbance, the biogas reactors were brought back to the original operational conditions; however, new steady-state microbial community profiles were observed in all the biogas reactors.ConclusionsThe present study demonstrated that temperature disturbance, but not stochastic factors, played an important role in shaping the profile of the microbial community composition and activity in biogas reactors. New steady-state microbial community profiles and reactor performances were observed in all the biogas reactors after the temperature disturbance.

Microplate-based method for high-throughput screening of microalgae growth potential

Microalgae cultivation conditions in microplates will differ from large-scale photobioreactors in crucial parameters such as light profile, mixing and gas transfer. Hence volumetric productivity (P(v)) measurements made in microplates cannot be directly scaled up. Here we demonstrate that it is possible to use microplates to measure characteristic exponential growth rates and determine the specific growth rate light intensity dependency (μ-I curve), which is useful as the key input for several models that predict P(v). Nannochloropsis salina and Chlorella sorokiniana specific growth rates were measured by repeated batch culture in microplates supplied with continuous light at different intensities. Exponential growth unlimited by gas transfer or self-shading was observable for a period of several days using fluorescence, which is an order of magnitude more sensitive than optical density. The microplate datasets were comparable to similar datasets obtained in photobioreactors and were used an input for the Huesemann model to accurately predict P(v).

Microbial analysis in biogas reactors suffering by foaming incidents

Foam formation can lead to total failure of digestion process in biogas plants. In the present study, possible correlation between foaming and the presence of specific microorganisms in biogas reactors was elucidated. The microbial ecology of continuous fed digesters overloaded with proteins, lipids and carbohydrates before and after foaming incidents was characterized using 16S rRNA gene sequencing. Moreover, the microbial diversity between the liquid and foaming layer was assessed. A number of genera that are known to produce biosurfactants, contain mycolic acid in their cell wall, or decrease the surface tension of the media, increased their relative abundance after foam formation. Finally, a microorganism similar to widely known foaming bacteria (Nocardia and Desulfotomaculum) was found to increase its relative abundance in all reactors once foam was observed, regardless of the used substrate. These findings suggest that foaming and specific microorganisms might have direct association which requires to be further investigated.

Effective harvesting of the microalgae Chlorella protothecoides via bioflocculation with cationic starch.

In the present work, the flocculation efficiency of cationic starch (Greenfloc 120) was tested on the fresh water microalga Chlorella protothecoides under different conditions (pH and flocculant concentrations). Different concentrations of Greenfloc 120 (0, 2.5, 5, 10, 20, 40 mg L(-1)) were screened against different algal densities (0.44, 0.56 and 0.77 g L(-1)). Once the optimal flocculation concentration had been established (40 mg L(-1) for all different biomasses densities) a more detailed analysis was performed in order to investigate if different pH (4.0, 7.7, and 10.0) could increase the flocculation efficiency of cationic starch. Highest flocculation efficiency without addition of Greenfloc 120 was obtained at pH 10, while in the presence of flocculant, the efficiency increased for all the tested pH values, with a maximum of 98% for pH 7.7 and 10. Cationic starch confirmed to be as an easy to use, efficient and cost-effective flocculant for harvesting of microalgae.

Effect of nitrogen source and acclimatization on specific growth rates of microalgae determined by a high-throughput in vivo microplate autofluorescence method

Specific growth rates (SGR) of freshwater algae species (Chlorella vulgaris, Auxenochlorella protothecoides, and Chlorella sorokiniana) and the marine species Nannochloropsis oculata on various nitrogen sources (ammonium carbonate, ammonium chloride, sodium nitrate, and urea) could be determined by in vivo chlorophyll-a autofluorescence. These preferences could be determined before large pH changes occurred in the media, with no significant difference (P > 0.05) between buffered and non-buffered media. In all algal species, acclimatization was observed with no significant difference (P > 0.05) between SGRs of the second and third cultivations. ANOVA of SGRs in the acclimatized second and third cultivations revealed preferences for nitrogen sources among most of the algae; C. vulgaris preferred sodium nitrate over other nitrogen sources, A. protothecoides adapted to urea after no growth in the first cultivation, and the SGRs of N. oculata showed an aversion for sodium nitrate over other nitrogen sources (P < 0.05).

Evaluation of microalgae production coupled with wastewater treatment

ABSTRACT In the present study, the feasibility of microalgae production coupled with wastewater treatment was assessed. Continuous cultivation of Chlorella sorokiniana with wastewater was tested in lab-scale flat-panel photobioreactors. Nitrogen and phosphorus removals were found to be inversely proportional to the four dilution rates, while chemical oxygen demand removal was found to be 50% at all the tested conditions. The biomass obtained at the highest dilution rate was characterized for its content of lipids, proteins and pigments. The average yields of fatty acid methyl esters (FAMEs), protein, lutein, chlorophylls and β-carotene was 62.4, 388.2, 1.03, 11.82 and 0.44 mg per gram dry biomass, respectively. Economic analysis revealed that potentially more than 70% of revenue was from the production of pigments, that is, chlorophyllin (59.6%), lutein (8.9%) and β-carotene (5.0%) while reduction in discharging costs of the treated wastewaters could account for 19.6% of the revenue. Due to the low market price of biodiesel, the revenue from the above was found to be the least profitable (1.4%). Even when combining all these different revenues, this cultivation strategy was found with the current prices to be uneconomical. Power consumption for artificial light was responsible for the 94.5% of the production costs.

The RNA polymerase subunits E/F from the Antarctic archaeon Methanococcoides burtonii bind to specific species of mRNA

RNA polymerase in Archaea is composed of 11 or 12 subunits - 9 or 10 that form the core, and a heterodimer formed from subunits E and F that associates with the core and can interact with general transcription factors and facilitate transcription. While the ability of the heterodimer to bind RNA has been demonstrated, it has not been determined whether it can recognize specific RNA targets. In this study we used a recombinant archaeal MbRpoE/F to capture cellular mRNA in vitro and a microarray to determine which transcripts it specifically binds. Only transcripts for 117 genes (4% of the total) representing 48 regions of the genome were bound by MbRpoE/F. The transcripts represented important genes in a number of functional classes: methanogenesis, cofactor biosynthesis, nucleotide metabolism, transcription, translation, import/export. The arrangement and characteristics (e.g. codon and amino acid usage) of genes relative to the putative origin of replication indicate that MbRpoE/F preferentially binds to mRNA of genes whose expression may be important for cellular fitness. We also compared the biophysical properties of RpoE/F from M. burtonii and Methanocaldococcus jannaschii, demonstrating a 50°C difference in their apparent melting temperatures. By using MbRpoE/F to capture and characterize cellular RNA we have identified a previously unknown functional property of the MbRpoE/F heterodimer.