Project description:The production of polyhydroxyalkanoates (PHAs) has been herein investigated by using an organic acid mixture originated from a pilot-scale acidogenic fermentation (AF) of reground pasta (RP) byproduct. The pilot-scale AF process was conducted either under no pH control or with the pH maintained at a value of 5.90, with the two obtained fermented mixtures termed RP-fermented 1 and RP-fermented 2, respectively. The fermented mixtures were fed to a lab-scale sequencing batch reactor (SBR), operated at short hydraulic retention time (HRT, 0.5 days) and sludge retention time (SRT, 1 day) and at two values of the applied organic loading rate (OLR) of 2.12 gCODACIDS/Ld and 4.25 gCODACIDS/Ld. During all of the SBR operating conditions, a high selective microbial pressure was established, as confirmed by both the microbiology analysis and the detected values of the storage yield (which reached a maximum value of 0.68 ± 0.04 CODPHA/CODACIDS). A poly(hydroxybutyrate/hydroxyvalerate) copolymer and a poly(hydroxybutyrate/hydroxyvalerate/hydroxyhexanoate) terpolymer were produced with the RP-fermented 1 and RP-fermented 2 streams, respectively. When the OLR of 2.12 gCODACIDS/Ld was applied to the SBR, the stored copolymer and terpolymer presented very similar molecular weights of 339 and 389 kDa, respectively.
Project description:This study compares the effects of pre- and post-hydrothermal treatment of source- separated organics (SSO) on solubilization of particulate organics and acidogenic fermentation for volatile fatty acids (VFAs) production. The overall COD solubilization and solids removal efficiencies from both schemes were comparable. However, the pre-hydrolysis of SSO followed by acidogenic fermentation resulted in a relatively higher VFA yield of 433 mg/g VSS, which was 18% higher than that of a process scheme with a post-hydrolysis of dewatered solids from the fermentation process. Regarding the composition of VFA, the dominance of acetate and butyrate was comparable in both process schemes, while propionate concentration considerably increased in the process with pre-hydrolysis of SSO. The microbial community results showed that the relative abundance of Firmicutes increased substantially in the fermentation of pretreated SSO, indicating that there might be different metabolic pathways for production of VFAs in fermentation process operated with pre-treated SSO. The possible reason might be that the abundance of soluble organic matters due to pre-hydrolysis might stimulate the growth of more kinetically efficient fermentative bacteria as indicated by the increase in Firmicutes percentage.
Project description:Biological sulfate reduction (BSR) represents a promising strategy for bioremediation of sulfate-rich waste streams, yet the impact of metabolic interactions on performance is largely unexplored. Here, genome-resolved metagenomics was used to characterize 17 microbial communities in reactors treating synthetic sulfate-contaminated solutions. Reactors were supplemented with lactate or acetate and a small amount of fermentable substrate. Of the 163 genomes representing all the abundant bacteria, 130 encode 321 NiFe and FeFe hydrogenases and all genomes of the 22 sulfate-reducing microorganisms (SRM) encode genes for H2 uptake. We observed lactate oxidation solely in the first packed bed reactor zone, with propionate and acetate oxidation in the middle and predominantly acetate oxidation in the effluent zone. The energetics of these reactions are very different, yet sulfate reduction kinetics were unaffected by the type of electron donor available. We hypothesize that the comparable rates, despite the typically slow growth of SRM on acetate, are a result of the consumption of H2 generated by fermentation. This is supported by the sustained performance of a predominantly acetate-supplemented stirred tank reactor dominated by diverse fermentative bacteria encoding FeFe hydrogenase genes and SRM capable of acetate and hydrogen consumption and CO2 assimilation. Thus, addition of fermentable substrates to stimulate syntrophic relationships may improve the performance of BSR reactors supplemented with inexpensive acetate.
Project description:BackgroundCo-fermentation is an attractive technology for improving volatile fatty acids (VFAs) production by treatment of solid organic wastes. However, it remains unclear how the composition of different organic matters in solid waste influences the VFAs distribution, microbial community structure, and metabolic pathway during acidogenic co-fermentation. In this study, different organic wastes were added into waste activated sludge (WAS) as co-fermentation substrates to explore the impact of organic matter composition on VFAs pattern and the microbiological mechanism .ResultsAcetate was the most dominant VFA produced in all fermentation groups, making up 41.3-57.6% of the total VFAs produced during acidogenic co-fermentation under alkaline condition. With the increased addition of potato peel waste, the concentrations of propionate and valerate decreased dramatically, while ethanol and butyrate concentrations increased. The addition of food waste caused gradual decreases of valerate and propionate, but ethanol increased and butyrate was relatively stable. Some inconsistency was observed between hydrolysis efficiency and acidification efficiency. Our results revealed that starch was mainly responsible for butyrate and ethanol formation, while lipids and protein favored the synthesis of valerate and propionate. Microbial community analysis by high-throughput sequencing showed that Firmicutes had the highest relative abundance at phylum level in all fermentation groups. With 75% potato peel waste or 75% food waste addition to WAS, Bacilli (72.2%) and Clostridia (56.2%) were the dominant respective classes. In fermentation using only potato peel waste, the Bacilli content was 64.1%, while the Clostridia content was 53.6% in the food-only waste fermentation.ConclusionsAcetate was always the dominant product in acidogenic co-fermentation, regardless of the substrate composition. The addition of carbon-rich substrates significantly enhanced butyrate and ethanol accumulation, while protein-rich substrate substantially benefited propionate and valerate generation. Potato peel waste substantially favored the enrichment of Bacilli, while food waste dramatically increased Clostridia content in the sludge.
Project description:Valuable routes for sewage sludge utilization: effect of temperature and hydraulic retention time in the acidogenic fermentation process
| PRJNA830363 | ENA
Project description:Saline soil microbiome study biases caused by different DNA extraction protocols
Project description:Feeding with high-concentrate diets increases the risk of subacute ruminal acidosis (SARA). This experiment was conducted to evaluate whether supplementing a phytogenic feed additive based on L-menthol, thymol, eugenol, mint oil (Mentha arvensis) and cloves powder (Syzygium aromaticum) (PHY) can amend the ruminal fermentation profile, modulate the risk of SARA and reduce inflammation in cattle. The experiment was designed as a crossover design with nine non-lactating Holstein cows, and was conducted in two experimental runs. In each run, cows were fed a 100% forage diet one week (wk 0), and were then transitioned stepwise over one week (0 to 65% concentrate, wk adapt.) to a high concentrate diet that was fed for 4 weeks. Animals were fed diets either with PHY or without (CON). The PHY group had an increased ruminal pH compared to CON, reduced time to pH < 5.8 in wk 3, which tended to decrease further in wk 4, reduced the ruminal concentration of D-lactate, and tended to decrease total lactate (wk 3). In wk 2, PHY increased acetate, butyrate, isobutyrate, isovalerate, and the acetate to propionate ratio compared to CON. Phytogenic supplementation reduced inflammation compared to CON in wk 3. Overall, PHY had beneficial effects on ruminal fermentation, reduced inflammation, and modulated the risk of SARA starting from wk 3 of supplementation.
Project description:This study is focused on the effects of pH on the production of volatile fatty acids (VFAs) and their distribution through the acidogenic fermentation of source-sorted organic fraction of municipal solid waste (OFMSW) from a mechanical-biological treatment (MBT) plant, and food waste (FW) from a university canteen. In semi-continuous lab-scale digesters using OFMSW at a hydraulic retention time (HRT) of 3.5 days under acidic conditions (pH 6.0), the VFA concentration in the effluent increased to 9.8-11.5 g L-1 (VS content of the feedstock between 4.2 and 5.2% w/w), while its individual VFA profiling was similar to the influent which was already pre-fermented (namely, C2 35-41%, C3 18-22%, C4 17-21%, and C5 9-12%). When working with the same conditions but using FW as feedstock, an effluent with a VFA concentration up to 11.5 g VFA L-1 (FW with a VS content of 5.5% w/w) and a stable distribution of C2 and C4 acids (up to 60.3% and 12.9%, respectively) but with very low quantities of C3 and C5 acids (lower than 1.8 and 2.7%, respectively) was obtained. Anaerobic batch tests using FW revealed that alkaline pH near 9 could lead to higher VFA production with high acetic acid content when compared to pH 6. In the semi-continuous fermenters working at alkaline conditions (pH 9.5-10) using OFMSW and FW, an enhanced solubilization of organic matter was registered with respect to the fermenters working under acidic conditions. This fact was not reflected in a higher VFA production when using OFMSW as feedstock, probably due to free ammonia inhibition, since OFMSW was mixed in the MBT plant with supernatant from anaerobic digestion of this biowaste. However, when using FW, alkaline conditions lead to an enhanced VFA production with respect to the reactor working under acidic conditions, being acetic acid the predominant product, which represented up to 91% of the VFA spectrum obtained.
Project description:BackgroundInteractions between microorganisms during specific steps of anaerobic digestion determine metabolic pathways in bioreactors and consequently the efficiency of fermentation processes. This study focuses on conversion of lactate and acetate to butyrate by bacteria of dark fermentation. The recently recognized flavin-based electron bifurcation as a mode of energy coupling by anaerobes increases our knowledge of anaerobic lactate oxidation and butyrate formation.ResultsMicrobial communities from dark fermentation bioreactors or pure culture of Clostridium butyricum are able to convert lactate and acetate to butyrate in batch experiments. The ability of C. butyricum to transform lactate and acetate to butyrate was shown for the first time, with ethanol identified as an additional end product of this process. A search for genes encoding EtfAB complexes and their gene neighbourhood in C. butyricum and other bacteria capable of lactate and acetate conversion to butyrate as well as butyrate-producers only and the lactate oxidiser Acetobacterium woodii, revealed that the Etf complexes involved in (i) lactate oxidation and (ii) butyrate synthesis, form separate clusters. There is a more extent similarity between Etf subunits that are involved in lactate oxidation in various species (e.g. A. woodii and C. butyricum) than between the different etf gene products within the same species of butyrate producers. A scheme for the metabolic pathway of lactate and acetate transformation to butyrate in C. butyricum was constructed.ConclusionsStudies on the conversion of lactate and acetate to butyrate by microbial communities from dark fermentation bioreactors or Clostridium butyricum suggest that a phenomenon analogous to cross-feeding of lactate in gastrointestinal tract also occurs in hydrogen-yielding reactors. A scheme of lactate and acetate transformation pathway is proposed, based on the example of C. butyricum, which employs flavin-based electron bifurcation. This process utilizes electron-transferring flavoprotein (Etf) complexes specific for (i) lactate oxidation and (ii) butyrate formation. Phylogenetic analysis revealed that such complexes are encoded in the genomes of other bacteria capable of lactate and acetate conversion to butyrate. These findings contribute significantly to our understanding of the metabolic pathways and symbiotic interactions between bacteria during the acidogenic step of anaerobic digestion.
Project description:An understanding of the integrated relationships among the principal cellular functions that govern the bioenergetic reactions of an organism is necessary to determine how cells remain viable and optimise their fitness in the environment. Urease is a complex enzyme that catalyzes the hydrolysis of urea to ammonia and carbonic acid. While the induction of urease activity by several microorganisms has been predominantly considered a stress-response that is initiated to generate a nitrogen source in response to a low environmental pH, here we demonstrate a new role of urease in the optimisation of cellular bioenergetics. We show that urea hydrolysis increases the catabolic efficiency of Streptococcus thermophilus, a lactic acid bacterium that is widely used in the industrial manufacture of dairy products. By modulating the intracellular pH and thereby increasing the activity of β-galactosidase, glycolytic enzymes and lactate dehydrogenase, urease increases the overall change in enthalpy generated by the bioenergetic reactions. A cooperative altruistic behaviour of urease-positive microorganisms on the urease-negative microorganisms within the same environment was also observed. The physiological role of a single enzymatic activity demonstrates a novel and unexpected view of the non-transcriptional regulatory mechanisms that govern the bioenergetics of a bacterial cell, highlighting a new role for cytosol-alkalizing biochemical pathways in acidogenic microorganisms.