Thermophilic Fermentation Research Articles

Effects of metronidazole on mesophilic and thermophilic fermentation: Biodegradation mechanisms, microbial communities, and reversibility

Metronidazole (MNZ), an antibiotic that is specifically used for the treatment of anaerobic infections, may inhibit anaerobic fermentation. This work was designed to understand the fate and effects of MNZ in mesophilic fermentation (MF) and thermophilic fermentation (TF), respectively. The results showed that the removal of MNZ mainly occurred via biodegradation, rather than adsorption, and that MNZ could be completely degraded by opening the imidazole ring. MFs were more strongly inhibited by MNZ than TFs. MNZ concentration increased from 0 to 25 mg/L, hydrogen yield (HY) decreased from 167.5 to 16.8 mL/g glucose (90.0% decrease), and butyrate yield almost completely disappeared in MFs, whereas in TFs, HY decreased only from 101.1 to 89.3 mL/g glucose (11.7% decrease), and ethanol yield increased by 39.8%. Illumina MiSeq sequencing analysis showed that MNZ reduced the abundance of hydrogen-producing bacteria. Furthermore, the inhibition of MNZ on anaerobic fermentation was reversible.

Sulfate- and pH-driven metabolic flexibility in sugarcane vinasse dark fermentation stimulates biohydrogen evolution, sulfidogenesis or homoacetogenesis

Dark fermentation of sugarcane vinasse can be used as a “cleaning” step to remove sulfate prior to methanogenesis because sulfidogenic conditions can be successfully established in parallel with biohydrogen production. Using a 22 central composite rotational design (CCRD) and response surface methodology (RSM), this study assessed the impacts of bicarbonate and sulfate availability on the establishment of sulfidogenesis in the thermophilic (55 °C) fermentation of vinasse in batch reactors, equally assessing the impacts on biohydrogen evolution. CCRD-RSM results indicated the favoring of biohydrogen production at the lowest sulfate and bicarbonate concentrations, whilst the opposite was observed for sulfidogenesis. Glycerol, lactate, and hydrogen were the preferential electron donors utilized by sulfate-reducing bacteria (SRB), whilst ethanol was markedly consumed only at high sulfate concentrations. SRB were inhibited by sodium when dosing excess NaHCO3 and Na2SO4. Complementary tests revealed maximum biohydrogen production (2.40 mmol) out of the CCRD, at pH exceeding 7.5 with no interference of sulfidogenesis. Non-efficient biohydrogen production was observed at low pH (<5.0; ∼1.90 mmol) because the uptake of lactate was inhibited. Meanwhile, homoacetogenesis was established under intermediate pH range (5.5–6.5), as revealed by the accumulation of acetate (up to 2.5 g L−1). 16S rRNA gene amplicon sequencing further revealed the genera Thermoanaerobacterium/Pseudoclostridium, Desulfotomaculum/Desulfohalotomaculum and Sporomusaceae/Moorella as the main biohydrogen-producing, sulfate-removing and biohydrogen-consuming (homoacetogens) microbial groups, respectively. Hence, using a single inoculum source, vinasse may provide a butyrate-rich (along with biohydrogen-rich biogas) or a sulfate-free and acetate-rich fermented effluent, depending mainly on proper pH control.

Metaproteomics reveals protein composition of multiple saccharifying enzymes in nongxiangxing daqu and jiangxiangxing daqu under different thermophilic temperatures

SummaryDaqu provides hydrolytic enzymes that saccharifying the macromolecular carbohydrate in fermenting grains. The process of saccharification is the premise of Baijiu production. But little is known about the protein composition associated with the activity of saccharifying enzymes in daqu. This study investigated related proteins between nongxiangxing daqu and jiangxiangxing daqu by metaproteomics. We present that the amylase system (α‐amylase, glucoamylase and α‐glucosidase) and cellulase system (cellobiohydrolases, endoglucanase and β‐glucosidase) are regarded as the main saccharifying enzymes. Combined with protein‐based taxonomic annotation, the key proteins involving in amylase system were mainly from Kroppenstedtia, Lichtheimia, Byssochlamys and Thermomyces, while Thermoascu contributed the most to cellulase system. Moreover, it was found that the up‐regulated amylase proteins at thermophilic fermentation period in jiangxiangxing daqu in comparison with nongxiangxing daqu were affiliated to Aspergillus, Rhizomucor, Byssochlamys and Thermomyces, along with Thermoascu contributing to cellulase system. These findings will provide clues for improving saccharification efficiency in Baijiu production.

PH buffering additives mitigate the inhibition of thermophilic dry methane fermentation

pH buffering additives mitigate the inhibition of thermophilic dry methane fermentation

Sewage sludge as carbon source for polyhydroxyalkanoates: a holistic approach at pilot scale level

In the present study, polyhydroxyalkanoates (PHA) production by mixed microbial cultures (MMC) has been carried out using thermally pre-treated excess thickened waste activated sludge (WAS), applying the feast-famine approach at pilot scale. The preliminary results of WAS fermentation conducted both in mesophilic and thermophilic conditions, in combination with a thermal pre-treatment at 70 °C for 48 h, highlighted how the thermal hydrolysis has a crucial role for the solubilization of the chemical oxygen demand (COD), which allows a significant increase of the final volatile fatty acid (VFA) concentration (roughly 8.5 g CODVFA/L). Since thermophilic fermentation after thermal pre-treatment was the best performing condition, it has been applied at pilot scale in order to routinely produce VFA as precursors for the following PHA synthesis. The selection and enrichment of PHA-producing biomass was successfully established and maintained in a Sequencing Batch Reactor (SBR) during the whole experimentation period, under short hydraulic retention time (HRT; 2 days) and medium-low organic loading rate (OLR; 2.0–2.2 g COD/L d). At the end of the production process an average PHA content of 53 ± 3 %w/w was achieved. The polymer was finally extracted and recovered from the biomass using traditional chloroform and sodium hypochlorite extraction and then characterized for the quantification of thermal properties (Tm = 156.8–160.9 °C) and molecular weight (Mv = 396–405 kDa). A final overall mass balance, usually poorly reported in the literature, has been also assessed, resulting in an overall yield of 56 g PHA per kg of volatile solids (VS).

Integrated meta-omics study on rapid tylosin removal mechanism and dynamics of antibiotic resistance genes during aerobic thermophilic fermentation of tylosin mycelial dregs

For efficient treatment of tylosin mycelial dregs (TMDs), rapid tylosin removal mechanism and dynamics of ARGs during TMDs fermentation were investigated using integrated meta-omics (genomics, metaproteomics and metabolomics) and qPCR approaches. The results showed that over 86% of tylosin was degraded on day 7 regardless of the type of bulking agents. The rapid removal of tylosin was mainly attributed to de-mycarose reaction (GH3) and esterase hydrolysis (C7MYQ7) of Saccharomonospora, and catalase-peroxidase oxidation of Bacillus (A0A077JB13). In addition, the moisture content and mobile genetic elements were vital to control the rebound of ARGs. The removal efficiency of antibiotic resistant bacteria (Streptomyces, Pseudomonas, norank_f__Sphingobacteriaceae, and Paenalcaligenes) and Intl1 (98.8%) in fermentation treatment TC21 with corncob as the bulking agent was significantly higher than that in other three treatments (88.3%). Thus, appropriate bulking agents could constrain the abundance of antibiotic resistant bacteria and Intl1, which is crucial to effectively reduce the resistance.

Mechanisms of nitrogen transformation driven by functional microbes during thermophilic fermentation in an ex situ fermentation system

In this study, we explored the pathways and mechanisms of nitrogen (N) transformation driven by functional microbes carrying key genes in an ex situ fermentation system (EFS). Temperature and N content were found to be the most important factors driving variation in bacterial and fungal communities, respectively; Bacillus became the most abundant bacteria and Batrachochytrium became the most abundant fungi. Co-occurrence network analysis showed that some bacteria including Halomonas, Truepera, and Gemmatimonas species carry genes that promote mineralization, nitrification, dissimilatory/assimilatory nitrate reduction, denitrification, anammox reactions, and N fixation. The maximum rate of total mineralization reached 136.60 μg N g−1 d−1. Functional microbes promoted various N conversion processes at different rates in the EFS, with levels increasing by at least 0.23 μg N g−1 d−1. These results provide a theoretical basis for feasible optimization measures to address N loss during fermentation.

Plant biomass fermentation by the extreme thermophile Caldicellulosiruptor bescii for co-production of green hydrogen and acetone: Technoeconomic analysis

A variety of chemical and biological processes have been proposed for conversion of sustainable low-cost feedstocks into industrial products. Here, a biorefinery concept is formulated, modeled, and analyzed in which a naturally (hemi)cellulolytic and extremely thermophilic bacterium, Caldicellulosiruptor bescii, is metabolically engineered to convert the carbohydrate content of lignocellulosic biomasses (i.e., soybean hulls, transgenic poplar) into green hydrogen and acetone. Experimental validation of C. bescii fermentative performance demonstrated 82% carbohydrate solubilization of soybean hulls and 55% for transgenic poplar. A detailed technical design, including equipment specifications, provides the basis for an economic analysis that establishes metabolic engineering targets. This robust industrial process leveraging metabolically engineered C. bescii yields 206 kg acetone and 25 kg H2 per metric ton of soybean hull, or 174 kg acetone and 21 kg H2 per metric ton transgenic poplar. Beyond this specific case, the model demonstrates industrial feasibility and economic advantages of thermophilic fermentation.

Enhanced semi-continuous hydrogen production by addition of microplastics under mesophilic and thermophilic fermentation

In this study, the influences of polyethylene terephthalate microplastics (PET MPs) on the semi-continuous hydrogen (H2) production and microbial community under mesophilic and thermophilic fermentation were investigated. Results indicated that PET MPs can enhance the semi-continuous H2 production performance, and the highest H2 production rates in mesophilic and thermophilic MPs supplemented groups were 4.85 and 3.6 L L−1 d−1, which were 21.6% and 63.6% higher than the control groups, respectively. Moreover, it was found that PET MPs could favor the growth of H2-producing bacteria and optimize the microbial community structure. In mesophilic MPs supplemented groups, the abundance of genus Lactococcus and Ethanoligenens increased from 20.77% and 6.7% to 38.69% and 10.78%, respectively. The abundance of genus Thermoanaerobacterium became the dominant bacteria in thermophilic fermentation, and its abundance increased from 9.02% to 69.38%. This work provides novel insights into the function of MPs in fermentative H2 production system.

Conversion of Food Waste into 2,3-Butanediol via Thermophilic Fermentation: Effects of Carbohydrate Content and Nutrient Supplementation.

Fermentation of food waste into 2,3-butanediol (2,3-BDO), a high-value chemical, is environmentally sustainable and an inexpensive method to recycle waste. Compared to traditional mesophilic fermentation, thermophilic fermentation can inhibit the growth of contaminant bacteria, thereby improving the success of food waste fermentation. However, the effects of sugar and nutrient concentrations in thermophilic food waste fermentations are currently unclear. Here, we investigated the effects of sugar and nutrients (yeast extract (YE) and peptone) concentrations on 2,3-BDO production from fermenting glucose and food waste media using the newly isolated thermophilic Bacillus licheniformis YNP5-TSU. When glucose media was used, fermentation was greatly affected by sugar and nutrient concentrations: excessive glucose (>70 g/L) slowed down the fermentation and low nutrients (2 g/L YE and 1 g/L peptone) caused fermentation failure. However, when food waste media were used with low nutrient addition, the bacteria consumed all 57.8 g/L sugars within 24 h and produced 24.2 g/L 2,3-BDO, equivalent to a fermentation yield of 0.42 g/g. An increase in initial sugar content (72.9 g/L) led to a higher 2,3-BDO titer of 36.7 g/L with a nearly theoretical yield of 0.47 g/g. These findings may provide fundamental knowledge for designing cost-effective food waste fermentation to produce 2,3-BDO.

Increased biological and cathodic hydrogen production using a novel integrated thermophilic fermenter and dual anion exchange membrane bioelectrochemical system

Many researchers are interested in utilizing renewable and sustainable energy made by exoelectrogenic bacteria during electrodialysis for the separation of minerals and organic matters from aqueous environments. The aim of this study was to develop a novel thermophilic fermenter and dual anion exchange membrane bioelectrochemical system for separating biohydrogen production inhibitors from the thermophilic fermenter and thereby increasing biological and cathodic hydrogen production by food waste and wastewater.•Using this innovative system the biohydrogen production inhibitors were separated and nutrients (for example ammonium), alkalinity, buffering capacity and pH were preserved in the bioreactor at the same time, led to higher biological and cathodic hydrogen production.

Lactic acid production from mesophilic and thermophilic fermentation of food waste at different pH

It is promising to recover lactic acid (LA) from fermentation of food waste (FW). In this study, pH and temperatures were investigated comprehensively to find their effects on LA fermentation, and microbial analyses were used to take insight to the variation of LA production. The results showed that mesophilic fermentation benefited hydrolysis and acidification, leading to a high yield of LA, while thermophilic conditions restricted other producers at low pH, leading to a high purity of LA. Lactobacillus amylolyticus was the main LA producer under thermophilic conditions, but Thermoanaerobacterium thermosaccharolyticum boomed at pH 5.0–6.0 and it converted LA partly to butyric acid. Simultaneously, Bacillus coagulans also increased and improved the optical purity (OP) of L-LA. From a series of this study, an operational condition of pH 5.5 and temperature of 52 °C would be potentially suitable for lactate fermentation of FW with high purity of 89%, while a stable LA production with an OP of 68% was achieved at 55 °C and pH 6.0.

Production of n-caproate using food waste through thermophilic fermentation without addition of external electron donors

The effectiveness of producing n-caproate from food waste without external electron donors (EDs) was investigated through batch and semi-continuous fermentation. The maximum concentration of n-caproate reached 10,226.28 mg COD/L during semi-continuous fermentation. The specificity for n-caproate was the highest at 40.19 ± 3.95%, and the soluble COD conversion rate of n-caproate reached up to 22.50 ± 1.09% at the end of batch fermentation. The production of n-caproate was coupled with the generation of lactate as an ED to facilitate chain elongation reactions. When lactate was used as the only substrate, n-butyrate (64.12 ± 20.11%) markedly dominated the products, instead of n-caproate (0.63 ± 0.07%). Microbial community analysis revealed that Caproiciproducens, Rummeliibacillus, and Clostridium_sensu_stricto_12 were the key genera related to n-caproate production. In addition to n-caproate, n-butyrate dominated the products in batch and semi-continuous fermentation with a maximum specificity of 47.59 ± 3.39%. Clostridium_sensu_stricto_7 was committed to producing n-butyrate from lactate.

Effect of Geobacillus toebii GT-02 addition on composition transformations and microbial community during thermophilic fermentation of bean dregs

Bean dregs can be prepared into organic fertilizer by microbial fermentation. Geobacillus toebii GT-02, which has promoting effect on bean dregs fermentation, was isolated from horse dung and it grows within a range of 40–75 °C and pH 6.50–9.50. The effectiveness of GT-02 addition on composition transformations and the microbial community in bean dregs thermophilic fermentation at 70 °C for 5 days was investigated (T1). Fermentation of bean dregs without GT-02 served as control (CK). The results showed that T1 (the germination index (GI) = 95.06%) and CK (GI = 86.42%) reached maturity (defined by GI ≥ 85%) on day 3 and day 5, respectively. In addition, the total nitrogen loss of T1 (18.46%) on day 3 was lower than that in CK (24.12%). After thermophilic fermentation, the total organic carbon and dry matter loss of T1 (53.51% and 54.16%) was higher than that in CK (41.72% and 42.82%). The mean microbial number in T1 was 4.94 × 107 CFUs/g dry matter, which was 5.37 times higher than that in CK. 16S rDNA sequencing identified Bacillus, Geobacillus and Thermobacillus as dominant in CK, while Bacillus, Ammoniibacillus and Geobacillus were dominant in T1. A canonical correspondence analysis showed that Geobacillus and Ammoniibacillus were positively correlated with the GI. Thus, thermophilic fermentation with GT-02 can promote the maturity of bean dregs, which indicated the potential application value of GT-02 in thermophilic fermentation.

Manganese ferrite nanoparticles enhanced biohydrogen production from mesophilic and thermophilic dark fermentation

In this work, Manganese ferrite nanoparticles (MnFe 2O4 NPs) were synthesized, and their potential to increase the biohydrogen (bio-H2) yield of glucose in mesophilic and thermophilic fermentations was evaluated. Material characterization showed that the MnFe 2O4 NPs were perfectly synthesized, with a specific surface area of 43.97 m2g−1. Both mesophilic and thermophilic fermentation obtained the highest bio-H2 yields of 272.7 and 183.4 mL H2/g glucose in the 400 mg/L MnFe 2O4 NPs group, which values were increased by 40.1% and 131.9% (37 °C and 55 °C), respectively, compared to their control groups. Soluble microbial products (SMPs) showed that MnFe 2O4 NPs enhanced the butyrate pathway. Three-dimensional fluorescence spectroscopy analysis showed that MnFe 2O4 NPs increased the extracellular polymer content of anaerobic sludge. The results showed that MnFe 2O4 NPs distinctly increased the abundance of Clostridium_sensu_stricto 1 (39.49%) in the microbial community during mesophilic fermentation and improved the abundance of the dominant bacteria Clostridium_sensu_stricto_7 (25.59%) and Longlinea (21.36%) during thermophilic fermentation. Real-time PCR analysis showed that hydrogenase gene (hycl) expression increased exponentially after the addition of MnFe 2O4 NPs, and increasing the fermentation temperature improved the hydrogenase activity. We found that MnFe 2O4 NPs promoted to bio-H2 production in both mesophilic and thermophilic fermentation, and it contributed more significantly to thermophilic fermentation than mesophilic fermentation.

Degradation of hop latent viroid during anaerobic digestion of infected hop harvest residues

The citrus bark cracking viroid (CBCVd) was identified as causal agent for a severe stunting disease in hops. Viroids are highly stable parasitic RNAs, which can be easily transmitted by agricultural practices. Since CBCVd has recently been detected in two European countries a growing concern is that this pathogen will further spread and thereby threaten the European hop production. Biogas fermentation is used to sanitize hop harvest residues infected with pathogenic fungi. Consequently, the aim of this study was to test if biogas fermentation can contribute to viroid degradation at mesophilic (40 °C) and thermophilic (50 °C) conditions. Therefore, a duplex reverse transcription real-time PCR analysis was developed for CBCVd and HLVd detection in biogas fermentation residues. The non-pathogenic hop latent viroid (HLVd) was used as viroid model for the pathogenic CBCVd. The fermentation trials showed that HLVd was significantly degraded after 30 days at mesophilic or after 5 days at thermophilic conditions, respectively. However, sequencing revealed that HLVd was not fully degraded even after 90 days. The incubation of hop harvest residues at different temperatures between 20 and 70 °C showed that 70 °C led to a significant HLVd degradation after 1 day. In conclusion, we suggest combining 70 °C pretreatment and thermophilic fermentation for efficient viroid decontamination.

ПЕРСПЕКТИВЫ ПОЛУЧЕНИЯ ГАЗОВОГО МОТОРНОГО ТОПЛИВА ИЗ БИОГАЗА АНАЭРОБНОГО СБРАЖИВАНИЯ ОТХОДОВ АПК

The ability of gas hydrates to concentrate gas into a solid chelate structure and the properties of self-preservation of gas hydrates at negative temperatures allows us to consider the possibility of developing a method for the utilization of biogas, environmentally safe storage and transportation of biomethane. (Research purpose) The research purpose is in substantiation the technological possibilities of obtaining synthetic mixed gas hydrates of biogas components, their storage and transportation based on the analysis of the existing regularities of the formation of gas hydrates in time, temperature and external pressure. (Materials and methods) The article presents the accumulated results of studies of the process of obtaining artificial hydrates of natural gas and methane- containing gas mixtures at various initial static pressures and temperatures. The object of research to substantiate the parameters of artificial creation of gas hydrates is biogas obtained during anaerobic thermophilic fermentation of organic waste at an existing experimental biogas plant. Mixed feed SK-8 with a humidity of 90-92 percent was used as an organic substrate of constant composition. The composition of biogas was studied using the Optima-7 Biogas gas analyzer. (Results and discussion) The article presents a process model and a technical appearance of an installation for producing gas motor fuel from the biogas of anaerobic digestion of organic waste of the agro-industrial complex. The hydrate formation time depends on the increase in the interfacial surface and the movement of gas bubbles relative to the liquid, which can be regulated by acting on the hydrate formation zone (shock wave, electromagnetic, mechanical, chemical, temperature). (Conclusions) The research results can be used in modeling processes in two-phase media during the formation of gas hydrates and the creation of installations for their production.

Cultured Milk

Animal milk use started between ca. 9000 and 6500 BCE at the onset of the Neolithic in Southwest Asia, and it subsequently accompanied the spread of farming into Europe from 6800/6700 BCE onward. Against the background of lambing seasons, climate, and technology, it is likely that prehistoric adults consumed not fresh milk but variants of lactose-reduced fermented milks and cheeses. While this is well in line with genetic lactose intolerance prevailing in the Neolithic, the strong selective force behind the regional emergence of genetic lactase persistence in later prehistory is still unknown. In a synopsis of the currently available archaeological data as well as knowledge of recent traditional dairying and microbiology, in this paper we trace the role of fermented dairy products in the prehistory of Southwest Asia and Europe, address possible spillover effects with other fermented products such as bread, beer, and wine, and outline working hypotheses for future research. Particular attention is drawn to the distinction between meso- and thermophilic fermentation as a possible clue as to the reasons for the strong selective advantage of lactase persistence.

Effect of hyperthermophilic pretreatment on methane and hydrogen production from garden waste under mesophilic and thermophilic conditions

Anaerobic digestion of garden waste was investigated in a two-stage process consisting of hyperthermophilic pretreatment followed by mesophilic or thermophilic fermentation. The greatest digestion performance was achieved when the substrates were first treated at 70 °C for 3 days with no inoculation, and then mixed with inoculum (anaerobic sludge) and subjected to anaerobic digestion at 55 °C. Under such conditions, the maximum methane and hydrogen yields from grass were 517 NmlCH4/kgVS and 52 NmlH2/kgVS, whereas the corresponding values for leaves were 421 NmlCH4/kgVS and 23 NmlH2/kgVS, and these figure were far greater than the yields obtained in experiments with no hyperthermophilic stage. A metagenomic analysis of hyperthermophilic environments revealed the appearance of thermophilic and hyperthermophilic bacteria showing hydrolytic activity against lignocellulosic materials, including Caldicellulosiruptor, Thermovenabulum, Thermoanaerobacter, Moorella, Tepimicrobium, Geobacillus and Thermobacillus at a genus level. A noticeable methane production in the hyperthermophilic stage could be linked to the presence of Methanothermobacter sp.

Effective Degradation of Cigarette Butts via Treatment with Old Landfill Leachates

In this paper, results of feasibility study on microplastics (MPs) assessment in leachates from the Latvian solid municipal landfill Getliņi are discussed. The application of leachates for the treatment of cigarette butts (CGB) was evaluated. Methods of fluorescent microscopy, Fourier transform infrared (FTIR) spectroscopy and FTIR-microscopy were used for the identification and characterization of MPs in the leachates and analysis of CGB. Presence of the secondary MPs (e.g., degraded polyolefin mixtures) was determined in the tested landfill leachates, while cellulose acetate (CA) was not determined in these products. The leachates were tested as potential media for the thermophilic (55°C) fermentation of CGB without air supply. Degradation of CGB was determined after one-week fermentation that was confirmed comparing the changes in FTIR spectra of CA prior and after the treatment. This study provoked a path for further experimental studies of controlled degradation of cigarette butts under natural conditions in landfill environments.