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Conversion of wastewater treatment plants into biorefineries is a sustainable alternative for obtaining valuable compounds, thus reducing pollutants and costs and protecting the environment and human health. Under specific operating conditions, microbial fermentative products of sewage sludge are volatile fatty acids (VFA) that can be precursors of polyhydroxyalkanoate thermoplastic polyesters. The role of various operating parameters in VFA production has yet to be elucidated. This study aimed to correlate the levels of VFA yields with prokaryotic microbiota structures of sewage sludge in two sets of batch fermentations with an initial pH of 8 and 10. The sewage sludge used to inoculate the batch fermentations was collected from a Sicilian WWTP located in Marineo (Italy) as a case study. Gas chromatography analysis revealed that initial pH 10 stimulated chemical oxygen demands (sCOD) and VFA yields (2020 mg COD/L) in comparison with initial pH 8. Characterization of the sewage sludge prokaryotic community structures—analyzed by next-generation sequencing of 16S rRNA gene amplicons—demonstrated that the improved yield of VFA paralleled the increased abundance of fermenting bacteria belonging to Proteobacteria, Bacteroidetes, Chloroflexi, and Firmicutes phyla and, conversely, the reduced abundance of VFA-degrading strains, such as archaeal methanogens.

The valorisation of protein-rich residual streams by anaerobic mixed-culture fermentation (MCF) has been barely studied in contrast to carbohydrate-rich wastes. The aim of this work was, therefore, to investigate how protein composition, i.e. the amino acid (AA) profile, affects the individual consumption of amino acids and, consequently, the outcome of the process. Mixed-culture fermentations were performed with two model proteins (casein and gelatin) using continuous and batch reactors at neutral pH values and 25 °C. The acidification was incomplete for both proteins, with casein achieving a higher value than gelatin. Albeit dominated by acetic acid, product spectra were different as well, with n-butyric acid as the second major product for casein and propionic acid for gelatin. The preferential consumption of amino acids was demonstrated, which interestingly depends on protein composition. The previously accepted stoichiometry accurately describes iso and n-butyric acid production, but it fails for propionic, iso and n-valeric acid generation. Overall, this study offers a better understanding of protein fermentation mechanisms, which will help to improve degradation models and to design fermentation processes, based on optimal substrate selection.

The objective of this study was to investigate the effects of 11 active compounds of essential oils (ACEO) on rumen fermentation characteristics and methane production. Two trials were conducted. In trial 1, ACEO (eugenol, carvacrol, citral, limonene, 1,4-cineole, p-cymene, linalool, bornyl acetate, α-pinene, and β-pinene) at a dose of 1,000 μL/L were incubated for 24 h in diluted rumen fluid with a 70:30 forage:concentrate substrate (16.2% crude protein; 36.6% neutral detergent fiber). Three fistulated Holstein cows were used as donors of rumen fluid. The reduction in methane production was observed with nine ACEO (up to 86% reduction) compared with the control (p

High-rate activated sludge (HRAS) can be used for volatile fatty acid (VFA) production prior or as an alternative to anaerobic digestion. The impact of fermentation pH, pre-treatment and hydraulic retention time (HRT) on the hydrolysis and VFA yields of HRAS sludge fermentation were investigated in continuous stirred tank reactors. Despite higher solubilization levels at pH 10 (38.3 % vs 23.4 %), VFA yields were similar at pH 8 (239 ± 14 mg COD-1 g CODfed and 216 ± 10 mg COD-1 g CODfed for pH 10 and 8 respectively), both operated at an HRT of 18 days. The introduction of an in-line mechanical sludge treatment at pH 8 resulted in a 1.4 fold increased VFA yield. Finally a reduced HRT from 18 to 9 and 6 days did not impact VFA yields significantly (p = 0.183). Since HRT and pH have a major impact on the economic feasibility of acidogenic fermentation, we propose to conduct HRAS fermentation at pH 8 and an HRT of 6 days in order to minimize the dosage of chemicals and reduce the volume of the fermentation tank.

Simple Summary The rumen is well known as a natural bioreactor for highly efficient degradation of fibers, and rumen microbes play an important role on fiber degradation. Carbohydrates are fermented by a variety of bacteria in the rumen and transformed into volatile fatty acids (VFAs) by the corresponding enzymes. However, the content of forage in the diet affects the metabolism of cellulose degradation and VFA production. Therefore, we combine metabolism and metagenomics to explore the effects of High forage/concentrate diets and sampling time on enzymes and microorganisms involved in the metabolism of fiber and VFA in cow rumen. This study showed that propionate formation via the succinic pathway, in which succinate CoA synthetase (EC 6.2.1.5) and propionyl CoA carboxylase (EC 2.8.3.1) were key enzymes. Butyrate formation via the succinic pathway, in which phosphate butyryltransferase (EC 2.3.1.19), butyrate kinase (EC 2.7.2.7) and pyruvate ferredoxin oxidoreductase (EC 1.2.7.1) are the important enzymes. The microorganisms are mainly affected by diet and sampling time. Abstract The objectives of this study were to investigate the difference in the mechanism of VFAs production combined with macrogenome technology under different forage-to-concentrate ratios and sampling times. Six ruminally cannulated Holstein cows were used in a randomized complete block design. The high forage (HF) and high concentrate (HC) diets contained 70 and 35% dietary forage, respectively. The results showed that pH was affected by sampling time, at 4 h after feeding had lower value. Excepted for acetate, the VFAs was increased with forage decreased. Propionate formation via the succinic pathway, in which succinate CoA synthetase (EC 6.2.1.5) and propionyl CoA carboxylase (EC 2.8.3.1) were key enzymes, and significantly higher in HC treatment than in HF treatment, Selenomonas, Ruminobacter, Prevotella, and Clostridium were the main microorganism that encodes these key enzymes. Butyrate formation via the succinic pathway, in which phosphate butyryltransferase (EC 2.3.1.19), butyrate kinase (EC 2.7.2.7) and pyruvate ferredoxin oxidoreductase (EC 1.2.7.1) are the important enzymes, Prevotella and Bacteroides played important role in encodes these key enzymes. This research gave a further explanation on the metabolic pathways of VFAs, and microorganisms involved in VFAs production under different F:C ration, which could further reveal integrative information of rumen function.

The aim of the present experiment was to study changes in volatile fatty acid (VFA) production using an isotope dilution technique, and changes in VFA fractional absorption rate (k aVFA) using a buffer incubation technique (BIT) during the dry period and early lactation, as affected by the postpartum (pp) rate of increase of concentrate allowance. The current results are complementary to previously reported changes on rumen papillae morphology from the same experiment. From 50 d antepartum to 80 d pp, VFA production rate was measured 5 times and k aVFA was measured 10 times in 12 rumen-cannulated Holstein Friesian cows. Cows had free access to a mixed ration, consisting of grass and corn silage, soybean meal, and (dry period only) chopped straw. Treatment consisted of either a rapid (RAP; 1.0 kg of DM/d; n = 6) or gradual (GRAD; 0.25 kg of DM/d; n = 6) increase of concentrate allowance (up to 10.9 kg of DM/d), starting at 4 d pp, aimed at creating a contrast in rumen-fermentable organic matter intake. For the BIT, rumen contents were evacuated, the rumen washed, and a standardized buffer fluid introduced [120 mM VFA, 60% acetic (Ac), 25% propionic (Pr), and 15% butyric (Bu) acid; pH 5.9 and Co-EDTA as fluid passage marker]. For the isotope dilution technique, a pulse-dose of 13C-labeled Ac, Pr, and Bu and Co-EDTA as fluid passage marker was infused. The rate of total VFA production was similar between treatments and was 2 times higher during the lactation (114 mol/d) than the dry period (53 mol/d). Although papillae surface area at 16, 30, and 44 d pp was greater in RAP than GRAD, Bu and Ac production at these days did not differ between RAP and GRAD, whereas at 16 d pp RAP produced more Pr than GRAD. These results provide little support for the particular proliferative effects of Bu on papillae surface area. Similar to developments in papillae surface area in the dry period and early lactation, the k aVFA (per hour), measured using the BIT, decreased from 0.45 (Ac), 0.53 (Pr) and 0.56 (Bu) at 50 d antepartum to 0.28 (Ac), 0.34 (Pr) and 0.38 (Bu) at 3 d pp. Thereafter, k aVFA (/h) rapidly increased up to 0.67 (Ac), 0.79 (Pr), and 0.79 (Bu) at 80 d pp. Although papillae surface area was greater at 16, 30, and 44 d pp in RAP than GRAD, no differences in k aVFA between RAP and GRAD were observed during these days showing papillae surface area is not the limiting factor for k aVFA during early pp adaptation.

The objective of this study was to investigate the effects of 11 active compounds of essential oils (ACEO) on rumen fermentation characteristics and methane production. Two trials were conducted. In trial 1, ACEO (eugenol, carvacrol, citral, limonene, 1,4-cineole, p-cymene, linalool, bornyl acetate, α-pinene, and β-pinene) at a dose of 1,000 μL/L were incubated for 24 h in diluted rumen fluid with a 70:30 forage:concentrate substrate (16.2% crude protein; 36.6% neutral detergent fiber). Three fistulated Holstein cows were used as donors of rumen fluid. The reduction in methane production was observed with nine ACEO (up to 86% reduction) compared with the control (p