Bioenergy recovery and carbon emissions benefits of short-term bio-thermophilic pretreatment on low organic sewage sludge anaerobic digestion: A pilot-scale study.

Sewage sludge in cities of Yangzi River Belt, China, generally exhibits a lower organic content and higher silt contentdue to leakage of drainage system, which caused low bioenergy recovery and carbon emission benefits in conventional anaerobic digestion (CAD). Therefore, this paper is on a pilot scale, a bio-thermophilic pretreatment anaerobic digestion (BTPAD) for low organic sludge (volatile solids (VS) of 4%) was operated with a long-term continuous flow of 200 days. The VS degradation rate and CH4 yield of BTPAD increased by 19.93% and 53.33%, respectively, compared to those of CAD. The analysis of organic compositions in sludge revealed that BTPAD mainly improved the hydrolysis of proteins in sludge. Further analysis of microbial community proportions by high-throughput sequencing revealed that the short-term bio-thermophilic pretreatment was enriched in Clostridiales, Coprothermobacter and Gelria, was capable of hydrolyzing acidified proteins, and provided more volatile fatty acid (VFA) for the subsequent reaction. Biome combined with fluorescence quantitative polymerase chain reaction (PCR) analysis showed that the number of bacteria with high methanogenic capacity in BTPAD was much higher than that in CAD during the medium temperature digestion stage, indicating that short-term bio-thermophilic pretreatment could provide better methanogenic conditions for BTPAD. Furthermore, the greenhouse gas emission footprint analysis showed that short-term bio-thermophilic pretreatment could reduce the carbon emission of sludge anaerobic digestion system by 19.18%.

Effect of lysozyme combined with hydrothermal pretreatment on excess sludge and anaerobic digestion.

Anaerobic digestion (AD) is widely employed for sludge stabilization and waste reduction. However, the slow hydrolysis process hinders methane production and leads to prolonged sludge issues. In this study, an efficient and eco-friendly lysozyme pre-treatment method was utilized to address these challenges. By optimizing lysozyme dosage, hydrolysis and cell lysis were maximized. Furthermore, lysozyme combined with hydrothermal pretreatment enhanced overall efficiency. Results indicate that: (1) When lysozyme dosage reached 90 mg/g TS after 240 min of pretreatment, SCOD, soluble polysaccharides, and protein content reached their maxima at 855.00, 44.09, and 204.86 mg/L, respectively. This represented an increase of 85.87%, 365.58%, and 259.21% compared to the untreated sludge. Three-dimensional fluorescence spectroscopy revealed the highest fluorescence intensity in the IV region (soluble microbial product), promoting microbial metabolic activity. (2) Lysozyme combined with hydrothermal pretreatment significantly increased SCOD, soluble proteins, and polysaccharide release from sludge, reducing SCOD release time. Orthogonal experiments identified Group 3 as the most effective for SCOD and soluble polysaccharide release, while Group 9 released the most soluble proteins. The significance order of factors influencing SCOD, soluble proteins, and polysaccharide release is hydrothermal temperature > hydrothermal time > enzymatic digestion time.(3) The lysozyme-assisted hydrothermal pretreatment group exhibited the fastest release and the highest SCOD concentration of 8,135.00 mg/L during anaerobic digestion. Maximum SCOD consumption and cumulative gas production increased by 95.89% and 130.58%, respectively, compared to the control group, allowing gas production to conclude 3 days earlier.

Life Cycle Assessment and Net Energy Analysis of an Integrated Hydrothermal Liquefaction-Anaerobic Digestion of Single and Mixed Beverage Waste and Sewage Sludge.

The conversion of biomass to bioenergy is one of the approaches to creating a sustainable society. In this study, the life cycle assessment and the net energy analysis of converting mixed sewage sludge and beverage waste into bioenergy via a combined hydrothermal liquefaction-anaerobic digestion (HTL-AD) system was carried out. Primary sludge (PS), winery rose lees (RL), brewery Trub (BT), the mixture of brewery trub and primary sludge (BTPS) and the mixture of winery rose lees and primary sludge (RLPS) were the feedstocks considered. Efficient energy utilization [in form of net energy ratio (NER)], and environmental emissions were evaluated. The NER of BT (2.07) and RL (1.76) increased when mixed with PS (3.18) to produce BTPS (3.20) and RLPS (2.85). Also, the HTL phase of the combined HTL-AD system produced a greater NER than the AD phase in BT, BTPS, and PS and vice-versa in RL and RLPS. Six environmental impact categories were studied namely global warming, terrestrial acidification, ionizing radiation, terrestrial ecotoxicity, human carcinogenic toxicity, and human non-carcinogenic toxicity. RL produced the greatest environmental impact while BTPS produced the least impact, thus indicating the advantage of feedstock combination. This study shows that the combination of feedstocks for bioenergy production in an HTL-AD system does not only increase the quality and quantity of products but also increases the overall NER as well as reducting the environmental impacts. The study also proved that an integrated HTL-AD system is an energy efficient system with greater resource utilization and less environmental footprint than the constituent systems.

Syntrophic propionate degradation in anaerobic digestion facilitated by hydrochar: Microbial insights as revealed by genome-centric metatranscriptomics.

Propionate is a model substrate for studying energy-limited syntrophic communities in anaerobic digestion, and syntrophic bacteria usually catalyze its degradation in syntrophy with methanogens. In the present study, metagenomics and metatranscriptomics were used to study the effect of the supportive material (e.g., hydrochar) on the key members of propionate degradation and their cooperation mechanism. The results showed that hydrochar increased the methane production rate (up to 57.1%) from propionate. The general transcriptional behavior of the microbiome showed that both interspecies H2 transfer (IHT) and direct interspecies electron transfer (DIET) played essential roles in the hydrochar-mediated methanation of propionate. Five highly active syntrophic propionate-oxidizing bacteria were identified by genome-centric metatranscriptomics. H85pel, a member of the family Pelotomaculaceae, was specifically enriched by hydrochar. Hydrochar enhanced the expression of the flagellum subunit, which interacted with methanogens and hydrogenases in H85pel, indicating that IHT was one of the essential factors promoting propionate degradation. Hydrochar also enriched H162tha belonging to the genus of Thauera. Hydrochar induced the expression of genes related to the complete propionate oxidation pathway, which did not produce acetate. Hydrochar and e-pili-mediated DIET were enhanced, which was another factor promoting propionate degradation. These findings improved the understanding of metabolic traits and cooperation between syntrophic propionate oxidizing bacteria (SPOB) and co-metabolizing partners and provided comprehensive transcriptional insights on function in propionate methanogenic systems.

Elucidating the role of sub-thermophilic temperature and pre-hydrolyzation for effective upgrading scheme of old swine manure digesters.

Solids concentration, temperature, and digester configuration were subjected to biomethanation study to identify effective retrofitting schemes for old swine waste digesters. Batch assays were commenced to determine an appropriate scenario at 30-55 °C and total solids 1-3 %TS. Sub-thermophilic temperature (45 °C) was found desirable with an additional 11.1 % methane yield, while digestion at higher TS induced ammonium inhibition. Subsequent batch experiments lasted 72 hrs for hydrolytic-acidogenic assessment under various temperatures. Heating control at 45 °C and 55 °C for 24 hrs increased hydrolysis efficiency 4.6-5.7 folds above control but showed no significant difference (α = 0.05) between them. Limited heat supply from biogas engine dictated the continuous digestion study to operate pre-hydrolysis reactor at maximum temperature of 45 °C. The two-stage strategy demonstrated best overall performances at the sub-thermphilic combination, raising methane yield by 35.4 %. Next-Generation Sequencing indicated remarkable shifts in abundance and diversity, especially for hydrolytic organisms, which expanded from 54 to 70.2 % by sub-thermophilic temperature.

Characterization, stability, and curcumin bioaccessibility of buckwheat flower polysaccharide conjugate emulsion.

In this study, buckwheat flower polysaccharide conjugates (BFPCs) were synthesized and evaluated for their emulsification properties. The stability of BFPC-stabilized emulsions was assessed through particle size analysis, zeta potential measurements, microscopic observations, and rheological tests. Gum Arabic served as a control to compare BFPC's emulsifying efficacy across varying storage conditions, including exposure to metal ions, pH variations, and different heat treatment temperatures. Results showed that BFPC significantly lowered interfacial tension (16.2 mN/m) and effectively stabilized emulsions containing 60 wt% medium-chain triglycerides at a concentration of 1.0 wt%. Over a 20-day storage period, BFPC emulsions demonstrated robust resistance to heat (60-90°C), acidic conditions (pH 2.0-9.0), and ion concentrations (Na+, Ca2+). Moreover, in a high oil phase emulsion, BFPC enhanced the bioavailability of curcumin to 27.05%, markedly higher than the 7.10% observed without emulsification, underscoring its potential in nutrient delivery applications. PRACTICAL APPLICATION: Due to its excellent resistance, long-time emulsifying stability under different conditions, and its good effect in curcumin embedding, BFPC has a broad prospect and can be widely used under various conditions in food industry.

Bioaccessibility and Antioxidant Activity of Faba Bean Peptides in Comparison to those of Pea and Soy after In Vitro Gastrointestinal Digestion and Transepithelial Transport across Caco-2 and HT29-MTX-E12 Cells.

In this study, the transepithelial transport of bioactive peptides derived from faba bean flour gastrointestinal digestates was investigated, in vitro, using a Caco-2 and HT29-MTX-E12 coculture monolayer, in comparison to those of pea and soy. The profile of transported peptides was determined by mass spectrometry, and the residual antioxidant activity was assessed. The ORAC value significantly (p < 0.05) decreased after transepithelial transport (24-36% reduction) for all legumes, while the antioxidant activity in ABTS assay significantly (p < 0.05) increased, as shown by the EC50 decrease of 26-44%. Five of the nine faba bean peptides that crossed the intestinal cell monolayer exhibited antioxidant activity. Two of these peptides, TETWNPNHPEL and TETWNPNHPE, were further hydrolyzed by the cells' brush border peptidases to smaller fragments TETWNPNHP and TWNPNHPE. These metabolized peptides were synthesized, and both maintained high antioxidant activity in both ABTS (EC50 of 1.2 ± 0.2 and 0.4 ± 0.1 mM, respectively) and ORAC (2.5 ± 0.1 and 3.4 ± 0.2 mM of Trolox equivalent/mM, respectively) assays. These results demonstrated for the first time the bioaccessibility of faba bean peptides produced after in vitro gastrointestinal digestion and how their bioactive properties can be modulated during transepithelial transport.

Biokinetic and microbial insights into regulatory mechanisms of long-chain fatty acid degradation during food waste-lipid co-digestion within anaerobic membrane bioreactor.

This study investigated the effects of varying lipid ratios on the anaerobic co-digestion of high-lipid food waste (FW) in a mesophilic anaerobic membrane bioreactor (AnMBR). At a lipid concentration of 5 %, optimal biogas production (3.84 L/L/d) and lipid removal efficiency (78 %) were achieved; however, increasing lipid concentrations resulted in significant accumulations of long-chain fatty acids (LCFAs) and volatile fatty acids (VFAs). Batch tests further demonstrated the impact of various types of LCFAs, with stearic acid showing the slowest microbial growth rate (0.033d-1), confirming its role in the accumulation of acetate-dominated VFAs, potentially limiting the methanogenesis process at elevated lipid levels. Furthermore, at 8 % lipid content, the downregulation of key LCFA degradation enzymes and dominance of hydrogenotrophic methanogens indicated adverse conditions. The importance of the intricate interplay between LCFA degradation kinetics and microbial community for the system efficiency was evidenced, offering insights for optimizing and managing high-lipidic wastes.

Two-stage solid-state fermentation to increase the nutrient value of corn processing waste and explore its efficacy as a feed protein source.

Corn gluten meal-corn husk mixes (CCM) are an inexpensive and readily available agricultural by-product. This study explores a novel technique by converting CCM into high-value livestock feed protein sources through fermentation with Aspergillus niger AAX and Lactobacillus fermentum LLS, aiming to sustainably meet future global protein needs. The process of fermentation significantly altered the structural composition of high molecular weight proteins, zein, and dietary fibers. This transformation resulted in a marked elevation in the concentrations of peptides, free amino acids, and polyphenols. The acidic environment produced during fermentation prevented lipid oxidation in CCM, thereby extending its storability. After fermentation, the content of anti-nutritional factors decreased, while its antioxidant capacity increased. In vitro simulated digestion suggested that fermentation improved the digestibility of CCM protein. In vivo animal experiments showed that fermented CCM (FCCM) promoted growth and gut health in chicks. This study provides new insights into the utilization of CCM.

Enhancing biogas production from cattle manure: A circular economy approach with solar thermal pretreatment and soil conditioning.

Biogas production from cattle manure, pivotal for sustainable waste and energy management, encounters challenges from its low digestibility linked to lignocellulosic structures. This study investigates biogas efficiency enhancement through anaerobic digestion coupled with solar thermal (ST) pretreatment and digestate application in Napier grass cultivation (STAD-G). ST pretreatment at 40-60 °C for 20 h markedly increases methane yield, validated by pilot-scale trials that exhibited a 3.9-fold surge in methane production, attributed to improved acidification. Untreated manure's structure, however, impedes acidification, slowing methanogenesis as shown by lower volatile fatty acid concentration in effluents. In addition, utilizing digestate as soil conditioner notably improves grass yield (19.3 ± 0.8 ton dry/ha/year) and protein content, akin to urea fertilizer. Furthermore, the STAD-G system incurs higher upfront costs, it yields superior biogas efficiency and enhanced long-term financial returns. This integrated approach, by boosting economic and environmental sustainability, advocates for ST pretreatment as a key strategy in advancing sustainable agriculture and energy solutions.

Antioxidant activity and inhibitory effects on angiotensin I-converting enzyme and α-glucosidase of trans-p-coumaroyl-secologanoside (comselogoside) and its inclusion complex with ß-cyclodextrin. Bioaccessibility during simulated in vitro gastrointestinal digestion.

This study explored the impact of complexing comselogoside (COM) with ß-cyclodextrin (ß-CD) on antioxidant capacity and investigated its in vitro inhibitory effects against α-glucosidase and angiotensin I-converting enzyme (ACE). The COM: ß-CD complex in three molar ratios (1:2, 1:1, and 2:1) showed significantly higher antioxidant activity compared to free COM, assessed by DPPH and ferric reducing power assays. COM exhibited weak to moderate α-glucosidase inhibition (IC50 1221 µM) and notable ACE inhibition (IC50 119.4 µM). Encapsulation improved ACE inhibition notably for the 1:2 and 2:1 M ratios. The cleavage of secoiridoid moiety of COM by ß-glucosidase further enhanced ACE inhibition from IC50 of 63.91 to 41.75 µg/mL in the hydrolysed mixture. In vitro gastrointestinal digestion revealed 34-40% bioaccessibility of COM and its ß-CD complex. This study demonstrates the potential of encapsulated COM as a functional food or supplement for preventing and treating diabetes, hypertension, and oxidative stress-related diseases.

Effect of different food matrices on the bioaccessibility of vitamin D3 in beverage systems: Comparison between juice and liquid milk.

Vitamin D plays a crucial role in bone, immunology, and neurophysiological functions but has inadequate bioavailability in the human body. In this paper, six different liquid beverages were used for vitamin D3 fortification, investigating the effect of different food matrices on the bioaccessibility of vitamin D. Not from concentrate (NFC) apple juice (9.34%) and NFC orange juice (8.12%) presented about 20% higher bioaccessibility of vitamin D3 than soybean and skim milk, and achieved a similar value of whole milk (8.04%). Meanwhile, the bioaccessibility of NFC apple and orange juice was markedly about 120% higher than that of apple clear juice. From the correlation analysis, the bioaccessibility of VD3 indicated significant correlations with small intestine retention (0.82) and viscosity (0.66). But small intestinal particle size showed a negative effect on bioaccessibility (-0.78). Therefore, food components, delivery matrices, and physicochemical properties of digesta were key factors to achieve higher bioaccessibility for guiding formulation design.

Insight into the performance and microbial community of anaerobic digestion treating cow manure with a novel iron-functionalized activated biochar.

The main objective of this research was to evaluate the impacts of FeCl3-activated biochar (FA-BC) on anaerobic digestion (AD) treating cow manure. The study focused on improving AD performance and understanding microbial community structure with the addition of FA-BC, while comparing FA-BC with other conductive additives, such as pristine biochar (P-BC), NaOH-activated biochar (NA-BC), and magnetite. Key findings indicated that FA-BC significantly enhanced the AD performance, supported by an increase in CH4 yield of 11-16% and a reduction in the lag phase by 51%. The high surface area and electrical conductivity of FA-BC synergistically facilitated direct interspecies electron transfer (DIET), leading to these improvements. On contrast, P-BC and NA-BC were not efficient in enhancing the AD performance due to relatively low electrical conductivity. P-BC also improved the CH4 yield, but less effectively than FA-BC. The effects of NA-BC varied with its dosage, showing inhibition at higher dosages due to excessive surface area. Magnetite, despite its high conductivity, made the limited enhancement in CH4 yield owing to its low surface area. Additionally, the statistical analyses revealed that each additive differently affected specific bacterial and archaeal groups depending on their physical and chemical properties. Thus, these findings suggest that FA-BC would be a highly promising additive for enhancing AD systems, with potential applications in waste management and renewable energy production.

A simplified non-reduced peptide mapping method with faster and efficient enzymatic digestion for characterization of native disulfide bonds in monoclonal and bispecific antibodies.

Development of monoclonal and bispecific antibody-based protein therapeutics requires detailed characterization of native disulfide linkages, which is commonly achieved through peptide mapping under non-reducing conditions followed by liquid chromatography-mass spectrometry (LC-MS) analysis. One major challenge of this method is incomplete protein digestion due to insufficient denaturation of antibodies under non-reducing conditions. For a long time, researchers have explored various strategies with the aim of efficiently digesting antibody drugs when the disulfide bonds remain intact, but few could achieve this by using a simple and generic approach with well controlled disulfide scrambling artifacts. Here, we report a simple method for fast and efficient mapping of native disulfides of monoclonal and bispecific antibody-based protein therapeutics. The method was optimized to achieve optimal digestion efficiency by denaturing proteins with 8 M urea plus 0-1.25 M guanidine-HCl at elevated temperature (50 °C), followed by two-step digestion with trypsin/Lys-C mix using a one-pot reaction. The only parameter that needs to be optimized for different proteins is the concentration of guanidine-HCl present. This simplified sample preparation eliminated buffer exchange and can be completed within three hours. By using this new method, all native disulfide bonds were confirmed for these monoclonal and bispecific antibodies with high confidence. When compared with a commercial kit utilizing low-pH digestion condition, the new method demonstrated higher digestion efficiency and shorter sample preparation time. These results suggest this new one-pot-two-step digestion method is suitable for the characterization of antibody disulfide bonds, particularly for those antibodies with digestion-resistant domains under typical digestion conditions.

Production and simulated digestion of high-load beads containing Schizochytrium oil encapsulated utilizing prilling technique.

The oil from the heterotroph Schizochytrium is a rich source of n-3 PUFA, particularly DHA, and therefore highly susceptible to oxidation. The present work reports the first application of coaxial prilling for the protection of this oil through microencapsulation. After process optimization, core-shell microparticles were produced with calcium or zinc alginate at different concentrations. Encapsulates were analyzed in their tocopherol and PUFA content. Prilling lowered the earlier but had little effect on the latter. Microcapsules coated with calcium alginate (1 % and 1.75 %) had higher oil load and encapsulation efficiency and were therefore submitted to in vitro digestion together with a simulated meal. Digesta were also analyzed with HPLC-qTOF and 1H NMR and compared to undigested encapsulates. While 1 % calcium shell granted lower oil release and protection from oxidation in the simulated gastrointestinal tract, chromatographic and spectroscopic data of digesta showed higher presence of lipid digestion products.

Potential prebiotic effects of soy by-products as novel dietary fibre: Structure, function, in vitro simulation of digestion and fermentation properties.

This study aimed to prepare soybean dregs dietary fibre (DF) using physically assisted chemical (KHMSO) modification and study its structure, function and vitro simulation experiments. The soluble dietary fibre (SDF) content in KHMSO increased and insoluble dietary fibre (IDF) content decreased. The modified DF surface becomes irregular and rough, and the results of XPS fitting indicated that the DF structure had different peak-splitting groups. The KHMSO-treated group had the lowest digestion rate in gastric fluid and the highest digestibility in intestine fluid. The OD600 of fecal cultures was increased to 0.915, and the increased abundance of microbiota was associated with the metabolism of SCFAs, such as Lachnospiraceae, as well as the higher n-butyric acid in the KHMSO-treated group compared to the other groups and lower than the inulin, suggesting KHMSO might enhance the production of functional foods aimed at promoting intestinal health.

Biomethanization of rigid packaging made entirely of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate): Mono- and co-digestion tests and microbial insights.

This study evaluates the anaerobic mesophilic mono- and co-digestion of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) plastic bottles as a proxy for rigid packaging materials. Initial tests showed a 97.3 ±â€¯0.2 % reduction in weight and an observable alteration in the surface (thinning, color fading and pitting) of the PHBH bottles after eight weeks. Subsequent tests showed that PHBH squares (3 × 3 cm) produced 400 NmL-CH4/g-VSfed, at a slower rate compared to powdered PHBH but with similar methane yield. Co-digestion experiments with food waste, swine manure, or sewage sludge showed successful digestion of PHBH alongside organic waste (even at a high bioplastic loading of 20 % volatile solids basis), with methane production comparable to or slightly higher than that observed in mono-digestion. Molecular analyses suggested that the type of co-substrate influenced microbial activity and that methane production was mainly driven by hydrogenotrophic methanogenesis. These results suggest the potential for integrating rigid PHBH packaging into anaerobic digesters.

Acute physiological effects on macromolecule digestion following oral ingestion of the enzyme blend Elevase® in individuals that had undergone an ileostomy, but were otherwise healthy-a randomized, double blinded, placebo-controlled exploratory study.

Digestive enzymes can selectively degrade proteins, carbohydrates and lipids; and their supplementation alongside food may accelerate the breakdown of complex food matrices, facilitate greater nutrient absorption, decrease food sensitivities and aid in the management of certain disease states. Several intrinsic and extrinsic factors govern food digestion and for every individual this phenomenon is unique. This study was conducted as a randomized, crossover, placebo-controlled design where each participant served as their own control. This post-hoc analysis investigated the impact of a dietary enzyme supplementation blend known as Elevase® on dietary macromolecule digestion in samples from otherwise healthy participants that had previously undergone a small bowel resection, resulting in an ileostomy (NCT04489810). This is the first time this study-paradigm has been used for the assessment of in vivo dietary breakdown following enzyme supplementation. Arguably, this technique offers superior data when compared to that generated in artificial gut digestion models, preclinical animal models, or indeed conventional clinical studies using stool analyses, as it allows real-time access to samples in situ in the small intestine where the majority of nutritional absorption takes place. It was demonstrated that after 4 h, Elevase® significantly increased monosaccharide levels (predominantly glucose and fructose) in the ileostomy samples taken from the same individuals on the same diet on a different day. In addition, the bile salt taurohyodeoxycholic acid was also increased, suggesting a physiological host response to the macromolecule digestion induced by the enzymatic blend. Overall, these findings suggest Elevase® could accelerate food digestion and potentially increase nutrient availability from the diet.

Evaluation of the effect of roasting and digestion on biological activity of compounds of coffee extracts - in vitro assessment of the bioavailability, cytoprotective properties and modulation of inflammatory response.

Coffee is the most consumed beverage in the world. Consumption of phenolic compounds present in coffee protects the body against oxidative stress generation, inflammatory response, and cancer development. The aim of the study was evaluation of biological activity of coffee extracts (obtained from green, as well as light and dark roasted Robusta and Arabica beans) and isolated fractions on human colon adenocarcinoma Caco-2 cells, which are used as a cellular model of intestinal barrier in bioavailability studies. Additionally, impact of coffee phenolics on oxidative stress level and anti-inflammatory activity has been studied with RAW 264.7 macrophages used in immunomodulatory research. It was demonstrated that the coffee constituents protection against oxidative stress, lipotoxicity and secretion of proinflammatory mediators is correlated with the presence of mono- and dichlorogenic acids and roasting process. It was demonstrated that coffee phytochemicals can decrease cells proliferation and bind to topoisomerase IIα being a dietary tool in cancer prevention.