Effects of the Interaction between Rumen Microbiota Density-VFAs-Hepatic Gluconeogenesis on the Adaptability of Tibetan Sheep to Plateau.

During the adaptive evolution of animals, the host and its gut microbiota co-adapt to different elevations. Currently, there are few reports on the rumen microbiota-hepato-intestinal axis of Tibetan sheep at different altitudes. Therefore, the purpose of this study was to explore the regulatory effect of rumen microorganism-volatile fatty acids (VFAs)-VFAs transporter gene interactions on the key enzymes and genes related to gluconeogenesis in Tibetan sheep. The rumen fermentation parameters, rumen microbial densities, liver gluconeogenesis activity and related genes were determined and analyzed using gas chromatography, RT-qPCR and other research methods. Correlation analysis revealed a reciprocal relationship among rumen microflora-VFAs-hepatic gluconeogenesis in Tibetan sheep at different altitudes. Among the microbiota, Ruminococcus flavefaciens (R. flavefaciens), Ruminococcus albus (R. albus), Fibrobactersuccinogenes and Ruminobacter amylophilus (R. amylophilus) were significantly correlated with propionic acid (p < 0.05), while propionic acid was significantly correlated with the transport genes monocarboxylate transporter 4 (MCT4) and anion exchanger 2 (AE2) (p < 0.05). Propionic acid was significantly correlated with key enzymes such as pyruvate carboxylase, phosphoenolpyruvic acid carboxylase and glucose (Glu) in the gluconeogenesis pathway (p < 0.05). Additionally, the expressions of these genes were significantly correlated with those of the related genes, namely, forkhead box protein O1 (FOXO1) and mitochondrial phosphoenolpyruvate carboxykinase 2 (PCK2) (p < 0.05). The results showed that rumen microbiota densities differed at different altitudes, and the metabolically produced VFA contents differed, which led to adaptive changes in the key enzyme activities of gluconeogenesis and the expressions of related genes.

Astragalus additive in feed improved serum immune function, rumen fermentation and the microbiota structure of early-weaned lambs.

AIM: The purpose of this study was to determine the mechanism of Astragalus activity on the immune function, rumen microbiota structure, and rumen fermentation of early-weaned lambs. METHODS AND RESULTS: Thirty healthy early-weaned lambs with similar body weights (17.42 ± 2.02 kg) were selected for the feeding experiment. The control group (KB) was fed a basal diet, and the Astragalus group (HQ) was fed 0.3% Astragalus additive on the basis of a basic diet. The formal trial period was 60 days. The results showed that the concentrations of blood immunoglobulin A (IgA) and immunoglobulin M (IgM) in the HQ group were significantly higher than those in the KB group (P < 0.05). Compared with the KB group, the concentrations of acetic acid, butyric acid, and total volatile fatty acids (VFAs) in the HQ group were higher (P < 0.01). The expression levels of the rumen epithelial-related genes MCT1, MCT4, NHE2, and ZO1 in the Astragalus group were significantly higher than those in the KB group (P < 0.05). 16S rRNA analysis showed that at the phylum level, Bacteroidetes in the HQ group significantly increased (P < 0.01); at the genus level, Prevotella (P < 0.01) and Succiniclasticum (P < 0.01) in the HQ group were found at significantly higher abundances than those in the KB group, and the results of microbiota gene and function prediction showed that "energy metabolism," "glycan biosynthesis and metabolic" pathways were significantly enriched in the HQ group (P < 0.05). CONCLUSION: As a feed additive, Astragalus can improve the immunity of early-weaned lambs, the structure of the rumen microbiota of lambs, and the fermentation capacity of the rumen.

A novel strategy and mechanism for high-quality volatile fatty acids production from primary sludge: Peroxymonosulfate pretreatment combined with alkaline fermentation.

To obtain high-quality VFAs production from primary sludge, a novel strategy that combined peroxymonosulfate (PMS) pretreatment and alkaline fermentation (i.e., PMS & pH9) was proposed in the study. The results showed that PMS & pH9 was efficient in sludge solubilization and hydrolysis, resulting in a maximal VFAs yield of 401.2 mg COD/g VSS, which was 7.3-, 2.1-, and 8.8-fold higher than the sole PMS, sole pH9, and control, respectively. Acetate comprised 87.6% of VFAs in this integration system. Mechanism investigations revealed that sulfate and free radicals produced by PMS play roles in improving VFAs yield under alkaline conditions. Besides, sulfate also aided in C3∼C5 VFAs converting to acetate under alkaline conditions depending on the increase of incomplete-oxidative sulfate-reducing bacteria (iso-SRB) (i.e., Desulfobulbus and Desulfobotulus). Moreover, the relative abundances of acid-forming characteristic genera (i.e., Proteiniborus, Proteinilcasticum, and Acetoanaerobium) were higher in PMS & pH9.

Evaluation of volatile fatty acids and ammonia recovery approach from landfill leachate using pilot-scale mechanical vapor recompression.

Treatment of landfill leachate is still a current problem due to the high treatment costs in addition to the difficulty of meeting the discharge criteria. However, there is a more important issue that should be underlined; it is also valuable compounds that leachate contains. Conventional methods used for treatment of leachate such as membrane filtration, advanced oxidation processes, biological processes and their combinations have largely focused on treatment. However, the recovery of ammonia and volatile organic acids (VFA) in leachate is a promising approach both to overcome high treatment costs and to sustainably manage leachate. In this study, leachate treatment potential was investigated by mechanical vapor recompression (MVR) process, which offers an operational opportunity to recover high value-added products from leachate while providing an effective treatment for wastewater. Optimum operating conditions for the pilot-scale MVR process have been determined by laboratory-scale studies. VFAs were recovered as organic acid salts from the pilot-scale MVR distillate, while ammonia recovery was accomplished as ammonium sulfate from a highly contaminated concentrate stream. VFA and ammonia recovery rates were 89% and 99%, respectively. The treatment cost of leachate with MVR process was calculated according to the data obtained in pilot scale MVR studies considering the operating cost, chemical cost and economical contribution of value-added products. The results showed that the integrated MVR-crystallization process, all treatment costs are covered, with a net gain of 3.8 USD/m3. Consequently, MVR integrated crystallization process offers an economical and sustainable solution for the treatment of leachate by recovering valuable products.

Thermo-alkaline pretreatment of excess sludge: Effects of temperature on volatile fatty acids accumulation and microbial community.

In order to explore the role of thermal-alkaline pretreatment temperatures (TAPT) in sludge fermentation and the microbial characteristics, five groups (100, 120, 140, 160 °C and control group) were set up and the results showed that the increasing TAPT promoted the dissolution of soluble chemical oxygen demand (SCOD) and VFAs, but had slight influence on the release of NH4+-N and PO43--P. What's more, when it was 120 °C, the SCOD dissolution was comparable to that at 160 °C. Overall, 120 °C was the optimal condition, corresponding to the fact that the maximum release of SCOD was 8788.74 mg/L (2.63 times of the control group), the maximum dissolution of VFAs was 4596 mg/L (about 1.28 times of the control group). The trend of C/N was not significant. High-throughput sequencing showed that Firmicutes and Actinobacteriota were enriched with the temperature increasing, while Proteobacteria and Chloroflexi did not change significantly. Firmicutes was in a stable dominant position. Temperature conditions brought about significant changes in microbial interspecific interaction. Carbohydrate and amino acids had the highest metabolic abundance, especially at 120 °C group. The change rule of amino acid metabolism was similar to that of lipid metabolism, and the abundance of energy metabolism gradually increased with temperature. The protein metabolism was greatly affected by temperature. This study revealed the effect of microbial mechanism of TAPT on the sludge acid production efficiency.

Emerging Strategies for Enhancing Propionate Conversion in Anaerobic Digestion: A Review.

Anaerobic digestion (AD) is a triple-benefit biotechnology for organic waste treatment, renewable production, and carbon emission reduction. In the process of anaerobic digestion, pH, temperature, organic load, ammonia nitrogen, VFAs, and other factors affect fermentation efficiency and stability. The balance between the generation and consumption of volatile fatty acids (VFAs) in the anaerobic digestion process is the key to stable AD operation. However, the accumulation of VFAs frequently occurs, especially propionate, because its oxidation has the highest Gibbs free energy when compared to other VFAs. In order to solve this problem, some strategies, including buffering addition, suspension of feeding, decreased organic loading rate, and so on, have been proposed. Emerging methods, such as bioaugmentation, supplementary trace elements, the addition of electronic receptors, conductive materials, and the degasification of dissolved hydrogen, have been recently researched, presenting promising results. But the efficacy of these methods still requires further studies and tests regarding full-scale application. The main objective of this paper is to provide a comprehensive review of the mechanisms of propionate generation, the metabolic pathways and the influencing factors during the AD process, and the recent literature regarding the experimental research related to the efficacy of various strategies for enhancing propionate biodegradation. In addition, the issues that must be addressed in the future and the focus of future research are identified, and the potential directions for future development are predicted.

Sex differences in rumen fermentation and microbiota of Tibetan goat.

BACKGROUND: The gut microbiota play an important role in maintaining host metabolism, the immune system and health, while sex, genotype, diet and health have specific effects on the composition of the gut microbiota. Therefore, to explore the sex differences in the structure and function of rumen microbiota in Tibetan goats, herein we analyzed sex differences in rumen fermentation parameters, rumen microbiota and the expression of genes related to VFA transport in Tibetan goats. RESULTS: The results showed that the contents of acetic acid and propionic acid in the rumen of TGM (Tibetan goat male) were significantly higher than those in TGFm (Tibetan goat female) (P < 0.05), and total VFAs was significantly higher in TGM than TGFm (P < 0.05). Expression of the VFA transport-related genes DRA, AE2, MCT-1, NHE1, and NHE2 in the rumen epithelium of TGFm was significantly higher than that in TGM. Analysis of the composition and structure of the rumen microbiota revealed significant sex differences. At the phylum level, Firmicutes and Bacteroidetes were the dominant phyla in Tibetan goats. In addition, Fibrobacteres and Spirochaetes had significantly greater relative abundances in TGFm than in TGM (P < 0.05). At the genus level, the relative abundance of Fibrobacter, Ruminococcus_1 and Pyramidobacter was significantly higher in TGFm than in TGM (P < 0.05). The functional prediction results showed that replication, recombination and repair, RNA processing and modification were mainly enriched in TGFm (P < 0.05). CONCLUSIONS: Correlation analysis revealed significant associations of some rumen microbiota with the fermentation product VFAs and VFA transport-related genes. We concluded that yearling TGM and TGFm have distinct fermentation and metabolism abilities when adapting to the plateau environment, which provides a certain sex reference basis for Tibetan goat adaptation to the plateau environment.

Comparison of primary and secondary sludge carbon sources derived from hydrolysis or acidogenesis for nitrate reduction and denitrification kinetics: Organics utilization and microbial community shift.

Seeking available and economical carbon sources for denitrification process is an intractable issue for wastewater treatment. However, no study compared different types of waste sludge as carbon source from denitrification mechanism, organics utilization and microbial community aspects. In this study, primary and secondary sludge were pretreated by thermophilic bacteria (TB), and its hydrolysis or acidogenic liquid were prepared as carbon sources for denitrification. At C/N of 8-3, the variations of NO3--N and NO2--N were profiled in typical cycles and denitrification kinetics was analyzed. Primary sludge achieved a competitive NOX-N removal efficiency with less dosage than secondary sludge. Fourier transform infrared (FTIR) spectroscopy was introduced to analyze organic composition from functional-group perspective and the utilization of organic matters in different sludge carbon sources was investigated. To further analyze the microbial community shift in different denitrification systems, high-throughput sequencing technology was applied. Results showed that denitrifier Thauera, belonging to Proteobacteria, was predominant, and primary sludge acidogenic liquid enriched Thauera most intensively with relative abundance of 47.3%.

Rumen Epithelial Development- and Metabolism-Related Genes Regulate Their Micromorphology and VFAs Mediating Plateau Adaptability at Different Ages in Tibetan Sheep.

The rumen is an important hallmark organ of ruminants and plays an important role in the metabolism and immune barrier of Tibetan sheep on the Plateau. However, there are few studies on rumen development and metabolism regulation in Tibetan sheep at different ages. Here, we comprehensively analyzed the immune function, fermentation function, rumen epithelial micromorphology and transcriptome profile of Tibetan sheep at different ages. The results showed that the concentration of IgG decreased and the concentration of IgM increased with age (p < 0.05), and the highest concentration of IgA was observed at 1.5 and 3.5 years of age. In terms of rumen fermentation characteristics, VFAs of 4-month-old lambs were the highest, followed by VFAs and NH3-N of Tibetan sheep at 3.5 years of age. Hematoxylin-eosin staining and transmission electron microscopy section examination of rumen epithelial tissue showed that the rumen papilla width increased with age (p < 0.001), the thickness of the stratum corneum decreased, the cells in the stratum corneum showed accelerated migration and the thickness of the rumen muscle layer increased (p < 0.001). Desmosomal junctions between the layers of rumen epithelium increased at 1.5 and 3.5 years old, forming a compact barrier structure, and the basal layer had more mitochondria involved in the regulation of energy metabolism. RNA-seq analysis revealed that a total of 1006 differentially expressed genes (DEGs) were identified at four ages. The DEGs of Tibetan sheep aged 4 months and 6 years were mainly enriched in the oxidation−reduction process and ISG15-protein conjugation pathway. The 1.5 and 3.5-year-olds were mainly enriched in skeletal muscle thin filament assembly, mesenchyme migration and the tight junction pathway. WGCNA showed that DEGs related to rumen microbiota metabolite VFAs and epithelial morphology were enriched in "Metabolism of xenobiotics by cytochrome P450, PPAR signaling pathway, Butanoate metabolism pathways" and participated in the regulation of rumen epithelial immune and fermentation metabolism functions of Tibetan sheep at different ages. This study systematically revealed the regulatory mechanism of rumen epithelial development and metabolism in the plateau adaptation of Tibetan sheep, providing a new approach for the study of plateau adaptation.

Dark fermentative volatile fatty acids production from food waste: A review of the potential central role in waste biorefineries.

Volatile fatty acids (VFAs) are high-value chemicals that are increasingly demanded worldwide. Biological production via food waste (FW) dark fermentation (DF) is a promising option to achieve the sustainability and environmental benefits typical of biobased chemicals and concurrently manage large amounts of residues. DF has a great potential to play a central role in waste biorefineries due to its ability to hydrolyze and convert complex organic substrates into VFAs that can be used as building blocks for bioproducts, chemicals and fuels. Several challenges must be faced for full-scale implementation, including process optimization to achieve high and stable yields, the development of efficient techniques for selective recovery and the cost-effectiveness of the whole process. This review aims to critically discuss and statistically analyze the existing relationships between process performance and the main variables of concern. Moreover, opportunities, current challenges and perspectives of a FW-based and fermentation-centred biorefinery layout are discussed.

Selective Separation of Acetic and Hexanoic Acids across Polymer Inclusion Membrane with Ionic Liquids as Carrier.

This paper first reports on the selective separation of volatile fatty acids (VFAs) (acetic and hexanoic acids) using polymer inclusion membranes (PIMs) containing quaternary ammonium and phosphonium ionic liquids (ILs) as the carrier. The affecting parameters such as IL content, VFA concentration, and the initial pH of the feed solution as well as the type and concentration of the stripping solution were investigated. PIMs performed a much higher selective separation performance toward hexanoic acid. The optimal PIM composed of 60 wt% quaternary ammonium IL with the permeability coefficients for acetic and hexanoic acid of 0.72 and 4.38 µm s-1, respectively, was determined. The purity of hexanoic acid obtained in the stripping solution increased with an increase in the VFA concentration of the feed solution and decreasing HCl concentration of the stripping solution. The use of Na2CO3 as the stripping solution and the involvement of the electrodialysis process could dramatically enhance the transport efficiency of both VFAs, but the separation efficiency decreased sharply. Furthermore, a coordinating mechanism containing hydrogen bonding and ion exchange for VFA transport was demonstrated. The highest purity of hexanoic acid (89.3%) in the stripping solution demonstrated that this PIM technology has good prospects for the separation and recovery of VFAs from aqueous solutions.

Anaerobic co-digestion of food waste/excess sludge: substrates - products transformation and role of NADH as an indicator.

The process of anaerobic co-digestion is vital importance to resource recovery from organic solid wastes such as food waste and municipal sludge. However, its application is hindered by the limited understanding on the complex substrates-products transformation reactions and mechanisms therein. In this study, food waste (FW) and excess sludge (ES) from municipal wastewater treatment were mixed at various ratios (ES/FW 5:0, 4:1, 2:1, 1:1, 1:2, 1:4, w/w), and the co-digestion process was studied in a batch test. The consumption of substrates including soluble proteins and carbohydrates, the variation in the intermediates such as various volatile fatty acids, and the production of hydrogen and methane gases were monitored. The results suggested that 4:1 was likely the optimal ratio where substrates were consumed and biogas generated efficiently, whereas 1:2 and 1:4 caused severe inhibition. Fermentation of ES alone produced mainly acetic and propionic acid, while the addition of FW led to butyric acid type fermentation. Intermediates in the fermentation liquid were tentatively identified, and the levels of NADH quantified using 3D-excitation/emission fluorescence spectrometry. One class of the intermediates, tryptophan-like proteins were correlated to the butyric acid accumulation in ES/FW mixtures, and NADH level was proposed as an indicator of VFAs production activities.

Age-Related Response of Rumen Microbiota to Mineral Salt and Effects of Their Interactions on Enteric Methane Emissions in Cattle.

Mineral salt bricks are often used in cow raising as compensation for mineral losses to improve milk yield, growth, and metabolic activity. Generally, effects of minerals are partially thought to result from improvement of microbial metabolism, but their influence on the rumen microbiota has rarely been documented to date. In this study, we investigated the response of microbiota to mineral salt in heifer and adult cows and evaluated ruminal fermentation and enteric methane emissions of cows fed mineral salts. Twelve lactating Holstein cows and twelve heifers fed a total mixed ration (TMR) diet were randomly allocated into two groups, respectively: a treatment group comprising half of the adults and heifers that were fed mineral salt and a control group containing the other half fed a diet with no mineral salt supplement. Enteric methane emissions were reduced by 9.6% (P < 0.05) in adults ingesting a mineral salt diet, while concentrations of ruminal ammonia, butyrate, and propionate were increased to a significant extent (P < 0.05). Enteric methane emissions were also reduced in heifers ingesting a mineral salt diet, but not to a significant extent (P > 0.05). Moreover, the concentrations of ammonia and volatile fatty acids (VFAs) were not significantly altered in heifers (P > 0.05). Based on these results, we performed high-throughput sequencing to explore the bacterial and archaeal communities of the rumen samples. Succiniclasticum and Prevotella, two propionate-producing bacteria, were predominant in samples of both adults and heifers. At the phylotype level, mineral salt intake led to a significant shift from Succiniclasticum to Prevotella and Prevotellaceae populations in adults. In contrast, reduced abundance of Succiniclasticum and Prevotella phylotypes was observed, with no marked shift in propionate-producing bacteria in heifers. Methanogenic archaea were not significantly abundant between groups, either in adult cows or heifers. The shift of Succiniclasticum to Prevotella and Prevotellaceae in adults suggests a response of microbiota to mineral salt that contributes to higher propionate production, which competes for hydrogen utilized by methanogens. Our data collectively indicate that a mineral salt diet can alter interactions of bacterial taxa that result in enteric methane reduction, and this effect is also influenced in an age-dependent manner.

Dynamics of intestinal metabolites and morphology in response to necrotic enteritis challenge in broiler chickens.

Despite the relatively small contribution to metabolizable energy that volatile fatty acids (VFAs) provide in chickens, these organic acids have been reported to play beneficial roles in the gastrointestinal tract (GIT) of birds, for example, inhibition of the growth of some pathogenic bacteria. However, information regarding the dynamics of these metabolites in the GIT of chickens is still scarce, especially under disease conditions such as necrotic enteritis (NE). Here, we investigated the dynamics of VFAs and lactic acid, and intestinal morphology in response to NE predisposing factors, that is, excessive dietary fishmeal and Eimeria inoculation, and causative agent Clostridium perfringens producing NetB toxin. The experiment was designed in a 2 × 2 × 2 factorial arrangement of treatments with or without: fishmeal feeding, Eimeria inoculation and C. perfringens challenge. The results showed that these factors significantly influenced composition and concentration of VFAs and lactic acids, pH and histomorphometry in one way or another. These changes may be important for the onset of NE or only the synergetic responses to micro environmental stress. Eimeria appeared to be more important than fishmeal in predisposing birds to NE, thus the application of Eimeria in NE challenge provides more consistent success in inducing the disease. The metabolic responses to various adverse factors such as excessive dietary fishmeal and Eimeria infection are complex. Thus, intensive efforts are required to better understand NE so as to achieve the control of the disease in the absence of antibiotics.

Community proteomics provides functional insight into polyhydroxyalkanoate production by a mixed microbial culture cultivated on fermented dairy manure.

Polyhydroxyalkanoates (PHAs) are bio-based, biodegradable polyesters that can be produced from organic-rich waste streams using mixed microbial cultures (MMCs). To maximize PHA production, MMCs are enriched for bacteria with a high polymer storage capacity through the application of aerobic dynamic feeding (ADF) in a sequencing batch reactor (SBR), which consequently induces a feast-famine metabolic response. Though the feast-famine response is generally understood empirically at a macro-level, the molecular level is less refined. The objective of this study was to investigate the microbial community composition and proteome profile of an enriched MMC cultivated on fermented dairy manure. The enriched MMC exhibited a feast-famine response and was capable of producing up to 40 % (wt. basis) PHA in a fed-batch reactor. High-throughput 16S rRNA gene sequencing revealed a microbial community dominated by Meganema, a known PHA-producing genus not often observed in high abundance in enrichment SBRs. The application of the proteomic methods two-dimensional electrophoresis and LC-MS/MS revealed PHA synthesis, energy generation, and protein synthesis prominently occurring during the feast phase, corroborating bulk solution variable observations and theoretical expectations. During the famine phase, nutrient transport, acyl-CoA metabolism, additional energy generation, and housekeeping functions were more pronounced, informing previously under-determined MMC functionality under famine conditions. During fed-batch PHA production, acetyl-CoA acetyltransferase and PHA granule-bound phasin proteins were in increased abundance relative to the SBR, supporting the higher PHA content observed. Collectively, the results provide unique microbial community structural and functional insight into feast-famine PHA production from waste feedstocks using MMCs.

Effect of initial total solids concentration on volatile fatty acid production from food waste during anaerobic acidification.

The effect of initial total solids (TS) concentration on volatile fatty acid (VFAs) production from food waste under mesophilic conditions (35 °C) was determined. VFAs concentration and composition, biogas production, soluble chemical oxygen demand concentration, TS and volatile solids (VS) reduction, and ammonia nitrogen [Formula: see text] release were investigated. The VFAs concentrations were 26.10, 39.68, 59.58, and 62.64 g COD/L at TS contents of 40, 70, 100, and 130 g/L, respectively. While the VFAs' yields ranged from 0.467 to 0.799 g COD/g VSfed, decreased as initial TS increased. The percentage of propionate was not affected by TS concentration, accounting for 30.19-34.86% of the total VFAs, while a higher percentage of butyrate and lower percentage of acetate was achieved at a higher TS concentration. Biogas included mainly hydrogen and carbon dioxide and the maximum hydrogen yield of 148.9 ml/g VSfed was obtained at 130 g TS/L. [Formula: see text] concentration, TS and VS reductions increased as initial TS increased. Considering the above variables, we conclude that initial TS of 100 g/L shall be the most appropriate to VFAs production.

Higher thermal remediation temperature facilitates the sequential bioaugmented reductive dechlorination.

The coupling of thermal remediation with microbial reductive dechlorination (MRD) has shown promising potential for the cleanup of chlorinated solvent contaminated sites. In this study, thermal treatment and bioaugmentation were applied in series, where prior higher thermal remediation temperature led to improved TCE dechlorination performance with both better organohalide-respiring bacteria (OHRB) colonization and electron donor availability. The 60 °C was found to be a key temperature point where the promotion effect became obvious. Amplicon sequencing and co-occurrence network analysis demonstrated that temperature was a more dominating factor than bioaugmentation that impacted microbial community structure. Higher temperature of prior thermal treatment resulted in the decrease of richness, diversity of indigenous microbial communities, and simplified the network structure, which benefited the build-up of newcoming microorganisms during bioaugmentation. Thus, the abundance of Desulfitobacterium increased from 0.11 % (25 °C) to 3.10 % (90 °C). Meanwhile, released volatile fatty acids (VFAs) during thermal remediation functioned as electron donors and boosted MRD. Our results provided temperature-specific information on synergistic effect of sequential thermal remediation and bioaugmentation, which contributed to better implementation of the coupled technologies in chloroethene-impacted sites.

Unraveling the effects and mechanisms of microplastics on anaerobic fermentation: Exploring microbial communities and metabolic pathways.

To investigate the effects of microplastics (MPs) on hydrolysis, acidification and microbial characteristics during waste activated sludge (WAS) anaerobic fermentation process, five different kinds of MPs were added into the WAS fermentation system and results indicated that, compared to the control group, the addition of polyvinyl chloride (PVC)-MPs exhibited the least inhibition on volatile fatty acids (VFAs), reducing them by 13.49 %. Conversely, polyethylene (PE)-MPs resulted in the greatest inhibition, with a reduction of 29.57 %. MPs, while accelerated the dissolution of WAS that evidenced by an increase of lactate dehydrogenase (LDH) release, concurrently inhibited the activities of relevant hydrolytic enzymes (α-Glucosidase, protease). For microbial mechanisms, MPs addition affected the proliferation of key microorganisms (norank_f_Bacteroidetes_vadinHA17, Ottowia, and Propioniclava) and reduced the abundance of genes associated with hydrolysis and acidification (pfkb, gpmI, ilvE, and aces). Additionally, MPs decreased the levels of key hydrolytic and acidogenic enzymes to inhibit hydrolysis and acidification processes. This research provides a basis for understanding and unveils impact mechanisms of the impact of MPs on sludge anaerobic fermentation.

Time-dependent interference of surfactants and CeO2/Fe2O3 nanoparticles co-occurrence on the volatile fatty acids biosynthesis during semi-continuous sludge fermentation.

Various exogenous contaminants typically coexist in waste activated sludge (WAS), and the long-term impacts of these co-occurring contaminants on WAS anaerobic fermentation and associated mechanisms remain largely unknown. This study reveals that the co-occurrence of surfactants and nanoparticles (NPs, i.e., Fe2O3 and CeO2, frequently detected in sludge) exhibited time-dependent impacts on the volatile fatty acids (VFAs) biosynthesis. Surfactants triggered WAS decomposition and enhanced NPs dispersion, leading to increased exposure of functional anaerobes to NPs toxicity, negatively affecting them. Consequently, key fermentation processes, acidogenic bacterial abundance, and metabolic functions were inhibited in co-occurrence reactors compared to those containing only surfactants in the early stage (before 56 d). Surprisingly, the fermentation systems containing surfactants collapsed subsequently, with VFAs yield at 72 d decreasing by 48.59-71.27 % compared to 56 d. The keystone microbes (i.e., Acidobacteria (16 d) vs Patescibacteria (56 d)) were reshaped, and metabolic traits (i.e., proB involved in intracellular metabolism) were downregulated by 0.05-78.02 % due to reduced microbial adaptive capacity (i.e., quorum sensing (QS)). Partial least squares path modeling (PLS-PM) analysis suggests that the microbial community was the predominant factor influencing VFAs generation. This study provides new insights into the long-term effects of co-contaminants on the biological treatment of WAS.

An integrated system combining Tetrasphaera-dominated enhanced biological phosphorus removal with sulfur autotrophic denitrification to enhance biological nutrients removal.

This study developed a two-stage process, including Tetrasphaera-dominated enhanced biological phosphorus-removal (EBPR(T)) sequencing batch reactor (SBR), followed by sulfur autotrophic denitrification (SADN) SBR, to achieve advanced nutrients removal from low VFAs wastewater. The removal efficiencies of nitrogen and phosphorus (PO43--P) reached 99 % with effluent PO43--P and total inorganic nitrogen (TIN) below 0.5 mg/L and 1 mg/L in EBPR(T) and SADN SBR, respectively. Mechanism analysis indicated that as increasing drainage ratio and complex carbon sources, free amino acids, glycogen, and PHA served as the endogenous carbon sources of Tetrasphaera to store energy. SADN contributed to approximately 80 % of nitrogen removal. DNA and cDNA results indicated Tetrasphaera was shifted from clade 2 to clade 1 after increasing the drainage ratio and the complexity of the carbon source, and Tetrasphaera (50.95 %) and Ca. Accumulibacter (9.12 %) were the most important functional microorganisms synergized to remove phosphorus at the transcriptional level in EBPR(T). Thiobacillus (45.97 %) and Sulfuritalea (9.24 %) were the dominant sulfur autotrophic denitrifiers at gene and transcriptional level in SADN. The results suggested that the EBPR(T) - SADN SBRs have great nutrient removal performance in treating low VFAs wastewater without additional carbon sources.