Cristiano Varrone

Microbial network for waste activated sludge cascade utilization in an integrated system of microbial electrolysis and anaerobic fermentation.

BackgroundBioelectrochemical systems have been considered a promising novel technology that shows an enhanced energy recovery, as well as generation of value-added products. A number of recent studies suggested that an enhancement of carbon conversion and biogas production can be achieved in an integrated system of microbial electrolysis cell (MEC) and anaerobic digestion (AD) for waste activated sludge (WAS). Microbial communities in integrated system would build a thorough energetic and metabolic interaction network regarding fermentation communities and electrode respiring communities. The characterization of integrated community structure and community shifts is not well understood, however, it starts to attract interest of scientists and engineers.ResultsIn the present work, energy recovery and WAS conversion are comprehensively affected by typical pretreated biosolid characteristics. We investigated the interaction of fermentation communities and electrode respiring communities in an integrated system of WAS fermentation and MEC for hydrogen recovery. A high energy recovery was achieved in the MECs feeding WAS fermentation liquid through alkaline pretreatment. Some anaerobes belonging to Firmicutes (Acetoanaerobium, Acetobacterium, and Fusibacter) showed synergistic relationship with exoelectrogens in the degradation of complex organic matter or recycling of MEC products (H2). High protein and polysaccharide but low fatty acid content led to the dominance of Proteiniclasticum and Parabacteroides, which showed a delayed contribution to the extracellular electron transport leading to a slow cascade utilization of WAS.ConclusionsEfficient pretreatment could supply more short-chain fatty acids and higher conductivities in the fermentative liquid, which facilitated mass transfer in anodic biofilm. The overall performance of WAS cascade utilization was substantially related to the microbial community structures, which in turn depended on the initial pretreatment to enhance WAS fermentation. It is worth noting that species in AD and MEC communities are able to build complex networks of interaction, which have not been sufficiently studied so far. It is therefore important to understand how choosing operational parameters can influence reactor performances. The current study highlights the interaction of fermentative bacteria and exoelectrogens in the integrated system.

Recent Advances and Challenges towards Sustainable Polyhydroxyalkanoate (PHA) Production

Sustainable biofuels, biomaterials, and fine chemicals production is a critical matter that research teams around the globe are focusing on nowadays. Polyhydroxyalkanoates represent one of the biomaterials of the future due to their physicochemical properties, biodegradability, and biocompatibility. Designing efficient and economic bioprocesses, combined with the respective social and environmental benefits, has brought together scientists from different backgrounds highlighting the multidisciplinary character of such a venture. In the current review, challenges and opportunities regarding polyhydroxyalkanoate production are presented and discussed, covering key steps of their overall production process by applying pure and mixed culture biotechnology, from raw bioprocess development to downstream processing.

Evaluation of surfactants on waste activated sludge fermentation by pyrosequencing analysis

The effects of three widely-used surfactants on waste activated sludge (WAS) fermentation and microbial community structures were investigated. Rhamnolipid bio-surfactants (RL) showed more positive effects on WAS hydrolysis and acidification compared to chemosynthetic surfactants, such as sodium dodecylsulphate (SDS) and sodium dodecyl benzene sulfonate (SDBS). The highest SCOD and VFAs concentrations obtained with RL were 1.15-fold and 1.16-fold that of SDS, and up to 1.73 and 3.63 times higher than those obtained with SDBS. Pyrosequencing analysis showed that an evident reduction in bacterial diversity in surfactant-treated WAS. Moreover, acid-producing bacteria (such as Megasphaera and Oscillibacter), detected with RL, were (6.8% and 6.4% in proportion) more abundant than with SDS, and were rarely found in SDBS and the control. The results also revealed that RL allowed efficient hydrolysis enhancement and was favorable to functional microorganisms for further acidification during WAS fermentation.

Methane production enhancement by an independent cathode in integrated anaerobic reactor with microbial electrolysis.

Anaerobic digestion (AD) represents a potential way to achieve energy recovery from waste organics. In this study, a novel bioelectrochemically-assisted anaerobic reactor is assembled by two AD systems separated by anion exchange membrane, with the cathode placing in the inside cylinder (cathodic AD) and the anode on the outside cylinder (anodic AD). In cathodic AD, average methane production rate goes up to 0.070 mL CH4/mL reactor/day, which is 2.59 times higher than AD control reactor (0.027 m(3) CH4/m(3)/d). And COD removal is increased ∼15% over AD control. When changing to sludge fermentation liquid, methane production rate has been further increased to 0.247 mL CH4/mL reactor/day (increased by 51.53% comparing with AD control). Energy recovery efficiency presents profitable gains, and economic revenue from increased methane totally self-cover the cost of input electricity. The study indicates that cathodic AD could cost-effectively enhance methane production rate and degradation of glucose and fermentative liquid.

Comparison of Different Strategies for Selection/Adaptation of Mixed Microbial Cultures Able to Ferment Crude Glycerol Derived from Second-Generation Biodiesel

Objective of this study was the selection and adaptation of mixed microbial cultures (MMCs), able to ferment crude glycerol generated from animal fat-based biodiesel and produce building-blocks and green chemicals. Various adaptation strategies have been investigated for the enrichment of suitable and stable MMC, trying to overcome inhibition problems and enhance substrate degradation efficiency, as well as generation of soluble fermentation products. Repeated transfers in small batches and fed-batch conditions have been applied, comparing the use of different inoculum, growth media, and Kinetic Control. The adaptation of activated sludge inoculum was performed successfully and continued unhindered for several months. The best results showed a substrate degradation efficiency of almost 100% (about 10 g/L glycerol in 21 h) and different dominant metabolic products were obtained, depending on the selection strategy (mainly 1,3-propanediol, ethanol, or butyrate). On the other hand, anaerobic sludge exhibited inactivation after a few transfers. To circumvent this problem, fed-batch mode was used as an alternative adaptation strategy, which led to effective substrate degradation and high 1,3-propanediol and butyrate production. Changes in microbial composition were monitored by means of Next Generation Sequencing, revealing a dominance of glycerol consuming species, such as Clostridium, Klebsiella, and Escherichia.

Vertical distribution of bacterioplankton in Lake Averno in relation to water chemistry.

The effects of environmental factors on bacterioplankton distribution along the water column of Lake Averno (Naples, Italy) have been investigated by means of denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR)-amplified 16S rRNA gene fragments, and multivariate analysis applied to molecular data and physico-chemical parameters. Bacterial richness, estimated from DGGE profiles, remains constant throughout the water column, whereas archaeal richness increases with depth. Moreover, archaeal richness was significantly correlated to most of the measured abiotic variables, whereas bacterial richness did not. Analysis of sequences from DGGE bands revealed that the dominant bacterial populations belong to Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, Bacteroidetes and Firmicutes, whereas sequences of the archaeal DGGE bands are affiliated to Methanomicrobiales and Methanosarcinales orders. Moreover, bacterial sequences affiliated to Proteobacteria and Bacteroidetes do not group closely to typical freshwater lineages/clades/tribes within these phyla. Bacterioplankton distribution along the water column was apparently correlated with the vertical gradient of physico-chemical parameters as a statistically significant relationship between most of them with grouping of specific taxonomic units was observed.

Combined polyhydroxyalkanoates (PHA) and 1,3-propanediol production from crude glycerol: Selective conversion of volatile fatty acids into PHA by mixed microbial consortia

Crude glycerol is an important by-product of the biodiesel industry, which can be converted into volatile fatty acids (VFA) and/or 1,3-propanediol (1,3-PDO) by fermentation. In this study, a selective conversion of VFA to polyhydroxyalkanoates (PHA) was attained while leaving 1,3-PDO in the supernatant by means of mixed microbial consortia selection strategies. The process showed highly reproducible results in terms of PHA yield, 0.99 ± 0.07 Cmol PHA/Cmol S (0.84 g COD PHA/g COD S), PHA content (76 ± 3.1 g PHA/100 g TSS) and 1,3-PDO recovery (99 ± 2.1%). The combined process had an ultimate yield from crude glycerol of 0.19 g COD PHA and 0.42 g COD 1,3-PDO per g of input COD. The novel enrichment strategy applied for selectively transforming fermentation by-products into a high value product (PHA) demonstrates the significance of the enrichment process for targeting specific bio-transformations and could potentially prove valuable for other biotechnological applications as well.

Polyhydroxyalkanoates (PHA) production from fermented crude glycerol: Study on the conversion of 1,3-propanediol to PHA in mixed microbial consortia

Crude glycerol, a by-product from the biodiesel industry, can be converted by mixed microbial consortia into 1,3-propanediol (1,3-PDO) and volatile fatty acids. In this study, further conversion of these main products into polyhydroxyalkanoates (PHA) was investigated with the focus on 1,3-PDO. Two different approaches for the enrichment of PHA accumulating microbial consortia using an aerobic dynamic feeding strategy were applied. With the first approach, where nitrogen was present during the whole cycle, no net production of PHA from 1,3-PDO was observed in the fermented effluent, not even in a nitrogen-limited PHA accumulation assay. Nevertheless, experiments in synthetic substrates revealed that the conversion of 1,3-PDO to PHA was possible under nitrogen limiting conditions. Thus, a different enrichment strategy was formulated where nitrogen was limited during the feast phase to stimulate the storage response. Nitrogen was still supplied during the famine phase. With the latter strategy, a net production of PHA from 1,3-PDO was observed at a yield of 0.24 Cmol PHA/Cmol 1,3-PDO. The overall yield from the fermented effluent was 0.42 Cmol PHA/Cmol substrate. Overall, the PHA yield from 1,3-PDO seemed to be limited, similarly to when using glycerol as a substrate, by a decarboxylation step and accumulation of other storage polymers such as glycogen, and possibly, lipid inclusions.

Dynamics of hydrogen-producing bacteria in a repeated batch fermentation process using lake sediment as inoculum

Abstract In this study, we evaluated the effectiveness of lake sediment as inoculum for hydrogen production through dark fermentation in a repeated batch process. In addition, we investigated the effect of heat treatment, applied to enrich hydrogen-producing bacteria, on the bacterial composition and metabolism. Denaturing gradient gel electrophoresis and molecular cloning, both performed using the 16S rDNA gene as target gene, were used to monitor the structure of the bacterial community. Hydrogen production and bacterial metabolism were analysed via gas chromatography and high-performance liquid chromatography. Both treated and non-treated inocula were able to produce high amounts of hydrogen. However, statistical analysis showed a clear difference in their bacterial composition and metabolism. The heat treatment favoured the growth of different Clostridia sp., in particular of Clostridium bifermentans, allowing the production of a constant amount of hydrogen over prolonged time. These cultures showed both butyrate and ethanol fermentation types. Absence of heat treatment allowed species belonging to the genera Bacillus, Sporolactobacillus and Massilia to outgrow Clostridia sp. with a reduction in hydrogen production and a significant metabolic change. Our data indicate that lake sediment harbours bacteria that can efficiently produce hydrogen over prolonged fermentation time. Moreover, we could show that the heat treatment stabilizes the bacterial community composition and the hydrogen production.

Experimental assessment of a process including microbial fuel cell for nitrogen removal from digestate of anaerobic treatment of livestock manure and agricultural wastes

aDipartimento di Ingegneria Chimica Materiali Ambiente, Sapienza University of Rome, Via Eudossiana 18, 00184, Rome, Italy. bDipartimento di Ingegneria Astronautica, Elettrica ed Energetica, Sapienza University of Rome, Via delle Sette Sale 12b, 00184, Rome, Italy cENEA, C.R. Casaccia. Via Anguillarese 301, 00123 Roma dAalborg University, Department of Chemistry and Bioscience, Section for Sustainable Biotechnology, Copenhagen, Denmark. luca.dipalma@uniroma1.it