Metagenomic insights into the microbiota involved in lactate and butyrate production and manipulating their synthesis in alfalfa silage.

AIMS: Lactate and butyrate are important indicators of silage quality. However, the microorganisms and mechanisms responsible for lactate and butyrate production in silage are not well documented. METHODS AND RESULTS: whole-metagenomic sequencing was used to analyse metabolic pathways, microbiota composition, functional genes, and their contributions to lactate and butyrate production in alfalfa silage with (SA) and without (CK) sucrose addition. Carbon metabolism was the most abundant metabolic pathway. We identified 11 and 2 functional genes associated with lactate and butyrate metabolism, respectively. Among them, D-lactate dehydrogenase (ldhA) and L-lactate dehydrogenase (ldhB) were most important for the transition between D/L-lactate and pyruvate and were primarily related to Lactobacillus in the SA group. The genes encoding L-lactate dehydrogenase (lldD), which decomposes lactate, were the most abundant and primarily associated with Enterobacter cloacae. Butyrate-related genes, mainly encoding butyryl-CoA: acetate CoA-transferase (but), were predominantly associated with Klebsiella oxytoca and Escherichia coli in the CK group. CONCLUSIONS: Enterobacteriaceae and Lactobacillaceae were mainly responsible for butyrate and lactate formation, respectively.

Metagenomic analysis reveals the linkages between bacteria and the functional enzymes responsible for potential ammonia and biogenic amine production in alfalfa silage.

AIMS: To clarify the molecular mechanisms underlying ammonia (NH3 ) and biogenic amines (BAEs) formation in alfalfa silage, whole metagenomic sequencing analysis was performed to identify the linkages between functional bacteria and their responsible enzymes in alfalfa silage prepared with and without sucrose addition. METHODS AND RESULTS: Genes encoding nitrite reductase (nirB) resulting in NH3 formation were the most abundant and were mostly assigned to Enterobacter cloacae and Klebsiella oxytoca. Putrescine-related genes, classified mainly to encode ornithine decarboxylase (odcA), were predominantly carried by Escherichia coli, Ent. cloacae and Citrobacter sp. Escherichia coli and Kl. oxytoca were the important species responsible for cadaverine and tyramine formation. Ent. cloacae, E. coli, and Kl. oxytoca dominated the bacterial community in naturally fermented alfalfa silage, whilst sucrose-treated silages greatly inhibited the growth of these species by promoting the dominance of Lactobacillus plantarum, thus decreasing the concentrations of NH3 , cadaverine, putrescine and tyramine. CONCLUSIONS: Enterobacteriaceae bacteria are mainly responsible for the NH3 , putrescine, cadaverine and tyramine formations in alfalfa silage. SIGNIFICANCE AND IMPACT OF THE STUDY: Whole metagenomic sequencing analysis served as a useful tool to identify the linkages between functional bacteria and associated enzymes responsible for NH3 and BAEs formation.

An efficient strategy to improve enzymatic hydrolysis of naked oat straw pretreated by Irpex lacteus.

The objective of this study was aiming at developing an efficient strategy to promote enzymatic hydrolysis of naked oat straw and deciphering the potential mechanism. Irpex lacteus and Phlebia acerina were employed to inoculated on the naked oat straw for 4 weeks which the changes of fiber components, fermentation losses, lignin-degrading enzymes production pattern were determined weekly. Furthermore, the 72 h enzymatic hydrolysis of ultimately fermented naked oat straw were also evaluated. The acid detergent lignin was degraded at about 25% along with the moderate dry matter and cellulose loss which both showed selective degradation. The lignin-degrading enzymes production patterns of the two fungi were different which lignin peroxidase was not detected in Irpex lacteus treatment. In addition, the activities of cellulolytic enzymes were higher in Phlebia acerina treatment. After 72 h enzymatic hydrolysis, the reducing sugar content and hydrolysis yield pretreated by Irpex lacteus was 12.92 g/L and 69.49%, respectively. It was much higher than that in sterilized substrate and Phlebia acerina treatment. Meanwhile, the hydrolysis yields of glucose, sum of xylose and arabinose were all improved by Irpex lacteus which were 30.96% and 25.62%, respectively, and showed significant enhancements compared to control and Phlebia acerina treatment. Irpex lacteus is one of effective white rot fungi which could promote the enzymatic hydrolysis of naked oat straw obviously.

Novel strategy to improve the colonizing ability of Irpex lacteus in non-sterile wheat straw for enhanced rumen and enzymatic digestibility.

Pretreatment with white rot fungi is a promising method to enhance the digestibility of lignocelluloses; however, sterilization of feedstocks prior to inoculation is one of the costliest steps. To improve the colonizing ability of white rot fungi under non-sterile condition, Irpex lacteus, Pleurotus ostreatus, and Phanerochaete chrysosporium were inoculated in the wheat straw ensiled for 28 days and incubated for 56 days to determine the changes in microbe counts, organic acid content, chemical composition, and rumen and enzymatic digestibility. Results showed that ensiling produced abundant organic acids and suppressed most microbes in wheat straw. Significant growth of I. lacteus was observed after 3 days of incubation, and molds were only detectable at day 7 in the group. At the end of incubation, aerobic bacteria and lactic acid bacteria decreased by 18% and 38% in the wheat straw treated with I. lacteus, but molds, aerobic bacteria, and lactic acid bacteria thrived in those treated with P. ostreatus and P. chrysosporium. Even more, P. ostreatus and P. chrysosporium increased the lignin content of the ensiled wheat straw by 34% and 65%. However, I. lacteus selectively degraded lignin by 28% and improved the rumen and enzymatic digestibility by 18% and 34%. The finding indicates that ensiling prior to fermentation with I. lacteus is an effective method to control spoilage microbes and to enhance the rumen and enzymatic digestibility of wheat straw.

A lower cost method of preparing corn stover for Irpex lacteus treatment by ensiling with lactic acid bacteria.

OBJECTIVE: This study investigated a method of preparing corn stover for Irpex lacteus (I. lacteus) treatment to improve its in vitro rumen degradability under non-sterile conditions. METHODS: Corn stover was inoculated with Lactobacillus plantarum (L. plantarum), Lactobacillus buchneri (L. buchneri), and an equal mixture of these strains, and ensiled for 0, 3, 7, 14, and 28 days. After each period, a portion of the silage was sampled to assess the silage quality, and another portion of the silage was further treated with I. lacteus at 28°C for 28 d. All the samples were analyzed for fermentation quality, chemical composition, and in vitro gas production (IVGP) as a measure of rumen fermentation capacity. RESULTS: Lactic acid bacteria (LAB) was found to improve the silage quality of the corn stover, and the corn stover silage inoculated with L. plantarum produced more lactic acid and higher IVGP than other silage groups. The I. lacteus colonies flourished in the early stage of corn stover silage, especially on the 3-d corn stover silage inoculated with both L. plantarum and L. buchneri. This led to an 18% decrease in the acid detergent lignin content, and a 49.6% increase in IVGP compared with the raw stover. CONCLUSION: The combination of ensiling with the mixed LAB inoculation and I. lacteus treatment provided a cost-effective method for the improvement of the IVGP of corn stover from 164.8 mL/g organic matter (OM) to 246.6 mL/g OM.

Characteristics of proteolytic microorganisms and their effects on proteolysis in total mixed ration silages of soybean curd residue.

OBJECTIVE: The objective of this study was to isolate proteolytic microorganisms and evaluate their effects on proteolysis in total mixed ration (TMR) silages of soybean curd residue. METHODS: TMRs were formulated with soybean curd residue, alfalfa or Leymus chinensis hay, corn meal, soybean meal, a vitamin-mineral supplement, and salt in a ratio of 25.0:40.0:30.0:4.0:0.5:0.5, respectively, on a basis of dry matter (DM). The microbial proteinases during ensiling were characterized, the dominate strains associated with proteolysis were identified, and their enzymatic characterization were evaluated in alfalfa (A-TMR) and Leymus chinensis (L-TMR) TMR silages containing soybean curd residue. RESULTS: Both A-TMR and L-TMR silages were well preserved, with low pH and high lactic acid concentrations. The aerobic bacteria and yeast counts in both TMR silages decreased to about 105 cfu g-1 FM (Fresh matter) and below the detection limit, respectively. The lactic acid bacteria count increased to 109 cfu g-1 FM. The total microbial proteinases activities reached their maximums during the early ensiling stage and then reduced in both TMR silages with fermentation prolonged. Metalloproteinase was the main proteinase when the total proteinases activities reached their maximums, and when ensiling terminated, metallo and serine proteinases played equally important parts in proteolysis in both TMR silages. Strains in the genera Curtobacterium and Paenibacillus were identified as the most dominant proteolytic bacteria in A-TMR and L-TMR, respectively, and both their proteinases were mainly with metalloproteinase characteristics. In the latter ensiling phase, Enterococcus faecium strains became the major sources of proteolytic enzymes in both TMR silages. Their proteinases were mainly of metallo and serine proteinases classes in this experiment. CONCLUSION: Proteolytic aerobic bacteria were substituted by proteolytic lactic acid bacteria during ensiling, and the microbial serine and metallo proteinases in these strains played leading roles in proteolysis in TMR silages.

Effect of Irpex lacteus, Pleurotus ostreatus and Pleurotus cystidiosus pretreatment of corn stover on its improvement of the in vitro rumen fermentation.

BACKGROUND: The present work investigated changes in corn stover pretreated with different white rot fungi. Corn stover was inoculated with Irpex lacteus, Pleurotus ostreatus and Pleurotus cystidiosus prior to incubation under solid-state fermentation conditions at 28 °C for 42 days. Changes in the chemical composition, in vitro rumen degradability, lignocellulolytic enzyme activity and multi-scale structure of the corn stover were analysed. RESULTS: Content of all lignocellulose components decreased to a certain extent after fungal pretreatment. The total gas production of sterilized corn stover treated with I. lacteus for 42 days increased from 200 to 289 mL g-1 organic matter. Moreover, the cellulase activity was highest at the later stage of I. lacteus pretreatment. Multi-scale structural analysis indicated that white rot fungal pretreatment, and in particular that of I. lacteus, increased and enlarged substrate porosity and caused changes in the structure of corn stover. CONCLUSION: Irpex lacteus pretreatment improved the nutritional value of corn stover as a ruminant feed by degrading both cellulose and acid-insoluble lignin as well as changing the structure of the cell walls. © 2018 Society of Chemical Industry.

Characterization of culturable yeast species associating with whole crop corn and total mixed ration silage.

OBJECTIVE: This study investigated the association of yeast species with improved aerobic stability of total mixed ration (TMR) silages with prolonged ensiling, and clarified the characteristics of yeast species and their role during aerobic deterioration. METHODS: Whole crop corn (WCC) silages and TMR silages formulated with WCC were ensiled for 7, 14, 28, and 56 d and used for an aerobic stability test. Predominant yeast species were isolated from different periods and identified by sequencing analyses of the 26S rRNA gene D1/D2 domain. Characteristics (assimilation and tolerance) of the yeast species and their role during aerobic deterioration were investigated. RESULTS: In addition to species of Candida glabrata and Pichia kudriavzevii (P. kudriavzevii) previously isolated in WCC and TMR, Pichia manshurica (P. manshurica), Candida ethanolica (C. ethanolica), and Zygosaccharomyces bailii (Z. bailii) isolated at great frequency during deterioration, were capable of assimilating lactic or acetic acid and tolerant to acetic acid and might function more in deteriorating TMR silages at early fermentation (7 d and 14 d). With ensiling prolonged to 28 d, silages became more (p<0.01) stable when exposed to air, coinciding with the inhibition of yeast to below the detection limit. Species of P. manshurica that were predominant in deteriorating WCC silages were not detectable in TMR silages. In addition, the predominant yeast species of Z. bailii in deteriorating TMR silages at later fermentation (28 d and 56 d) were not observed in both WCC and WCC silages. CONCLUSION: The inhibition of yeasts, particularly P. kudriavzevii, probably account for the improved aerobic stability of TMR silages at later fermentation. Fewer species seemed to be involved in aerobic deterioration of silages at later fermentation and Z. bailii was most likely to initiate the aerobic deterioration of TMR silages at later fermentation. The use of WCC in TMR might not influence the predominant yeast species during aerobic deterioration of TMR silages.

Effects of microbial enzymes on starch and hemicellulose degradation in total mixed ration silages.

OBJECTIVE: This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR) silage. METHODS: The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR) or Leymus chinensis hay (LTMR), corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. RESULTS: Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens), B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens) in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. CONCLUSION: The microbial amylase contributes to starch hydrolysis during the ensiling process in both TMR silages, whereas the microbial hemicellulase participates in the hemicellulose degradation only at the early stage of ensiling.

Dynamics Associated with Prolonged Ensiling and Aerobic Deterioration of Total Mixed Ration Silage Containing Whole Crop Corn.

This study investigated the dynamics associated with prolonged ensiling and aerobic deterioration of whole crop corn (WCC) silages and total mixed ration (TMR) silages containing WCC (C-TMR silages) to clarify the differences that account for the enhanced aerobic stability of TMR silages. Laboratory-scale barrel silos were randomly opened after 7, 14, 28, and 56 d of ensiling and were subjected to analyses of fermentation quality, microbial and temperature dynamics during aerobic exposure. WCC and C-TMR silages were both well preserved and microorganisms were inhibited with prolonged ensiling, including lactic acid bacteria. Yeast were inhibited to below the detection limit of 500 cfu/g fresh matter within 28 d of ensiling. Aerobic stability of both silages was enhanced with prolonged ensiling, whereas C-TMR silages were more aerobically stable than WCC silages for the same ensiling period. Besides the high moisture content, the weak aerobic stability of WCC silage is likely attributable to the higher lactic acid content and yeast count, which result from the high water-soluble carbohydrates content in WCC. After silo opening, yeast were the first to propagate and the increase in yeast levels is greater than that of other microorganisms in silages before deterioration. Besides, increased levels of aerobic bacteria were also detected before heating of WCC silages. The temperature dynamics also indicated that yeast are closely associated with the onset of the aerobic deterioration of C-TMR silage, whereas for WCC silages, besides yeast, aerobic bacteria also function in the aerobic deterioration. Therefore, the inclusion of WCC might contribute to the survival of yeast during ensiling but not influence the role of yeast in deterioration of C-TMR silages.

Fermentation characteristics and lactic Acid bacteria succession of total mixed ration silages formulated with peach pomace.

The objective of this study was to assess the use of peach pomace in total mixed ration (TMR) silages and clarify the differences in aerobic stability between TMR and TMR silages caused by lactic acid bacteria (LAB). The TMR were prepared using peach pomace, alfalfa hay or Leymus chinensis hay, maize meal, soybean meal, cotton meal, limestone, a vitamin-mineral supplement, and salt in a ratio of 6.0:34.0:44.4:7.0:5.0:2.5:1.0:0.1 on a dry matter (DM) basis. Fermentation quality, microbial composition, and the predominant LAB were examined during ensiling and aerobic deterioration. The results indicated that the TMR silages with peach pomace were well fermented, with low pH and high lactic acid concentrations. The aerobic stability of TMR silages were significantly higher than that of TMR. Compared with TMR silages with alfalfa hay, TMR silage with Leymus chinensis hay was much more prone to deterioration. Although the dominant LAB were not identical in TMR, the same dominant species, Lactobacillus buchneri and Pediococcus acidilactici, were found in both types of TMR silages after 56 d of ensiling, and they may play an important role in the aerobic stability of TMR silages.

Available sulfur and phosphorus transformation mechanism and functional microorganisms during sheep manure composting on Qinghai-Tibet Plateau under two moisture contents.

Understanding the mechanisms of sulfur and phosphorus transformation during composting is important for improving compost fertility. This study aims to investigate the microbial mechanism of available sulfur and phosphorus transformation during sheep manure composting under different moisture contents (45%: M45 and 60%: M60) on the Qinghai-Tibet Plateau using metagenomics technology. The results showed that the final available sulfur and phosphorus contents of M45 were 11% and 13% higher than those of M60, respectively. M45 enhanced sulfur oxidation, sulfate reduction, and thiosulfate disproportionation. These steps were significantly positively correlated with available sulfur, and Pseudomonas, Thermobifida, Luteimonas, Brevibacterium, Planifilum, and Xinfangfangia were the main participants. Available phosphorus was significantly positively correlated with polyphosphate degradation and inorganic P solubilization, and the main participants in these steps were Luteimonas, Brachybacterium, Corynebacterium, Jeotgalicoccus, Microbacterium, Streptomyces, and Pseudoxanthomonas. These findings reveal the microbial mechanisms of available and phosphorus transformation during composting at two moisture contents.

The maturity, humus content, and microbial metabolic function of sheep manure compost on the Qinghai-Tibet Plateau can be significantly improved by reducing the moisture content.

The Qinghai-Tibet Plateau (QTP) is characterized by an extreme hypoxia, which may lead to lack of sufficient oxygen for compost production, and thus seriously affecting the compost quality. The moisture content (MC) has a direct effect on the oxygen content of composting pile. At present, the research on the optimum moisture content of compost production on the QTP is still lacking. This study aimed to investigate the influences of MC on fermentation quality of sheep manure composting on the QTP and to further analyze the changes of microbial metabolic function and enzyme activity under different MC. Composting experiment with low MC (45%) and conventional MC (60%) was conducted in both summer and autumn. The results showed that the composting efficiency of 45% MC was better than 60% in both seasons, which was mainly manifested as longer high-temperature period (summer:16 d vs 14 d, autumn: 7 d vs 2 d), higher germination index (summer:136.1% vs 128.6%, autumn:103.5% vs 81.2%), and more humus synthesis (summer:159.8 g/kg vs 151.2 g/kg, autumn:136.1 k/kg vs 115.5 k/kg). The 45% MC can improve microbial metabolism, including increasing the abundance of functional genes involved in carbohydrate metabolism, amino acid metabolism, and nucleotide metabolism and improving the activities of cellulase, ß-glucosidase, protease, polyphenol oxidase, and peroxidase. In conclusion, 45% MC can improve the fermentation efficiency and products quality of sheep manure compost on QTP.

Enhanced lignin degradation by Irpex lacteus through expanded sterilization further improved the fermentation quality and microbial community during the silage preservation process.

Traditional autoclaving, slow degradation rate and preservation of biomass treated by fungi are the main factors restricting biological treatment. In our previous studies, strains with high efficiency and selective lignin degradation ability were obtained. To further solve the limiting factors of biological treatment, this paper proposed a composite treatment technology, which could replace autoclaves for fungal treatment and improve the preservation and utilization of fungal-pretreated straw. The autoclaved and expanded buckwheat straw were, respectively, degraded by Irpex lacteus for 14 days (CIL, EIL), followed by ensiling of raw materials (CK) and biodegraded straw of CIL and EIL samples with Lactobacillus plantarum for different days, respectively (CP, CIP, EIP). An expansion led to lactic acid bacteria, mold, and yeast of the samples below the detection line, and aerobic bacteria was significantly reduced, indicating a positive sterilization effect. Expansion before I. lacteus significantly enhanced lignin selective degradation by about 6%, and the absolute content of natural detergent solute was about 5% higher than that of the CIL. Moreover, EIL decreased pH by producing higher organic acids. The combination treatment created favorable conditions for ensiling. During ensiling, EIP silage produced high lactic acid about 26.83 g/kg DM and the highest acetic acid about 22.35 g/kg DM, and the pH value could be stable at 4.50. Expansion before I. lacteus optimized the microbial community for ensiling, resulting in EIP silage co-dominated by Lactobacillus, Pediococcus and Weissella, whereas only Lactobacillus was always dominant in CP and CIP silage. Clavispora gradually replaced Irpex in EIP silage, which potentially promoted lactic acid bacteria growth and acetic acid production. In vitro gas production (IVGP) in EIL was increased by 30% relative to CK and was higher than 24% in CIL. The role of expansion was more significant after ensiling, the IVGP in EIP was increased by 22% relative to CP, while that in CIP silage was only increased by 9%. Silage of fungal-treated samples reduced methane emissions by 28% to 31%. The study demonstrated that expansion provides advantages for fungal colonization and delignification, and further improves the microbial community and fermentation quality for silage, enhancing the nutrition and utilization value. This has practical application value for scaling up biological treatment and preserving the fungal-treated lignocellulose.

GM1 Ameliorates Neuronal Injury in Rats after Cerebral Ischemia and Reperfusion: Potential Contribution of Effects on SPTBN1-mediated Signaling.

Monosialoganglioside GM1 (GM1) has long been used as a therapeutic agent for neurological diseases in the clinical treatment of ischemic stroke. However, the mechanism underlying the neuroprotective function of GM1 is still obscure until now. In this study, we investigated the effects of GM1 in ischemia and reperfusion (I/R) brain injury models. Middle cerebral artery occlusion and reperfusion (MCAO/R) rats were treated with GM1 (60 mg·kg-1·d-1, tail vein injection) for 2 weeks. The results showed that GM1 substantially attenuated the MCAO/R-induced neurological dysfunction and inhibited the inflammatory responses and cell apoptosis in ischemic parietal cortex. We further revealed that GM1 inhibited the activation of NFκB/MAPK signaling pathway induced by MCAO/R injury. To explore its underlying mechanism of the neuroprotective effect, transcriptome sequencing was introduced to screen the differentially expressed genes (DEGs). By function enrichment and PPI network analyses, Sptbn1 was identified as a node gene in the network regulated by GM1 treatment. In the MCAO/R model of rats and oxygen-glucose deprivation and reperfusion (OGD/R) model of primary culture of rat cortical neurons, we first found that SPTBN1 was involved in the attenuation of I/R induced neuronal injury after GM1 administration. In SPTBN1-knockdown SH-SY5Y cells, the treatment with GM1 (20 µM) significantly increased SPTBN1 level. Moreover, OGD/R decreased SPTBN1 level in SPTBN1-overexpressed SH-SY5Y cells. These results indicated that GM1 might achieve its potent neuroprotective effects by regulating inflammatory response, cell apoptosis, and cytomembrane and cytoskeleton signals through SPTBN1. Therefore, SPTBN1 may be a potential target for the treatment of ischemic stroke.

Effects of turning frequency on fermentation efficiency and microbial community metabolic function of sheep manure composting on the Qinghai-Tibet Plateau.

This study explored the effects of turning frequency on fermentation efficiency and microbial metabolic function of sheep manure composting on the Qinghai-Tibet Plateau (QTP). Five treatments with different turning frequencies were set up in this study: turning every 1 day (T1), 2 days (T2), 4 days (T3), 6 days (T4), and 8 days (T5). Results showed that the high temperature period for T1 and T5 lasted only 4 days, while that for T2-T4 lasted more than 8 days. The germination index of T1 and T5 was lower than 80%, while that of T2-T4 was 100.6%, 97.8%, and 88.6%, respectively. This study further predicted the microbial metabolic function of T2-T4 using the bioinformatics tool PICRUSt2 (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) and determining the activities of various functional enzymes. The results showed that carbohydrate metabolism, protein metabolism, and nucleotide metabolism were the main metabolic pathways of microorganisms, and that T2 increased the abundance of functional genes of these metabolic pathways. The activities of protease, cellulase, and peroxidase in T2 and T3 were higher than those in T4, and the effect of T2 was more significant. In conclusion, turning once every 2 days can improve the quality of sheep manure compost on the QTP.

Mitigation of soil N2O emissions by decomposed straw based on changes in dissolved organic matter and denitrifying bacteria.

The return of decomposed straw represents a less explored potential option for reducing N2O emissions. However, the mechanisms underlying the effects of decomposed straw return on soil N2O mitigation are still not fully clear. Therefore, we used a helium atmosphere robotized continuous flow incubation system to compare the soil N2O and N2 emissions from four treatments: CK (control: no straw), WS (wheat straw), IWS (wheat straw decomposed with Irpex lacteus), and PWS (wheat straw decomposed with Phanerochaete chrysosporium). All the treatments have been fertilized with the same amount of KNO3. Furthermore, we also analyzed i) the chemodiversity of soil dissolved organic matter (DOM), ii) the nirS, nirK, and nosZ gene copies and relative abundances of denitrifying bacterial communities (DBCs), and iii) the specific linkages between N2O emissions and DOM and DBC. The results showed that the WS, IWS and PWS treatments increased N2O emissions compared to the CK treatment. However, applying decomposed straw to soil, especially straw treated with P. chrysosporium, effectively decreased the soil N2O and increased N2 emissions compared to WS and IWS. Moreover, the IWS and PWS treatments increased the CHO composition, but they decreased the CHON and CHOS compositions of heteroatomic compounds of DOM compared with the WS and CK treatments. Furthermore, the WS, IWS and PWS treatments all significantly increased the nirS and nosZ gene copies compared with the CK treatment. Additionally, compared with the other treatments, the PWS treatment significantly shaped the DBC and led to a higher relative abundance of Pseudomonas with nirS and nosZ genes. Meanwhile, Network analysis showed that the mitigation of N2O was closely related to particular DOM molecules, and specific DBC taxa. These results highlight the potential for decomposed straw amendments to mitigate of soil N2O emissions not only by changing soil DOM but also mediating the soil DBC.

Expansion Improved the Physical and Chemical Properties and In Vitro Rumen Digestibility of Buckwheat Straw.

The hard texture and poor palatability of straw are important factors that hinder its application in feed. Expansion is a technology that can improve the utilization of biomass, but few studies have comprehensively revealed how to change physicochemical characteristics to improve nutritional value. In this study, mechanical and chemical methods were combined to study the texture properties, rheological properties, and physicochemical structures of straw, and its utilization value was evaluated by in vitro rumen digestion. Expansion caused hemicellulose degradation, cellulose separation, and lignin redistribution, resulting in a decrease in crystallinity. The hardness and chewiness of expanded straw were reduced by 55% to 66%, significantly improving palatability. The compressive stress could be reduced by 54-73%, and the relaxation elasticity was reduced by 5% when expanded straw was compressed. The compression deformation of expanded straw was doubled compared to feedstock, and the compacting degree was improved. Expanded straw significantly improved digestibility and gas production efficiency, which was due to the pore structure increasing the attachment of rumen microorganisms; besides that, the reduction of the internal structural force of the straw reduced energy consumption during digestion. The lignin content decreased by 10%, the hardness decreased further in secondary expansion, but the digestibility did not improve significantly.

Effects of Regular Water Replenishment on Enzyme Activities and Fungal Metabolic Function of Sheep Manure Composting on the Qinghai-Tibet Plateau.

The dry climate characteristics of the Qinghai-Tibet Plateau will seriously affect microbial metabolism during composting. In this study, we aimed to investigate the effects of regular water supplementation on the fungal and enzymatic activities of sheep manure composting in the Qinghai-Tibet Plateau. The experiment set up the treatments of water replenishment once every 7 days(T2) and 3.5 days (T3) days, and no water supplementation was used as the control (T1). The results showed that regular water supplementation increased the activities of various enzymes during composting, and the activities of protease, cellulase, peroxidase and polyphenol oxidase in T3 were higher than those in T2. Regular water supplementation increased the relative abundance of Remersonia and Mycothermus, which were significantly positively correlated with the germination index, and degradation of organic components. Regular water supplementation could enrich fungi carbohydrate, protein, and nucleotide metabolisms, and T3 had a better effect. A redundancy analysis showed that environmental factors could significantly affect the fungal community; among them, moisture content (76.9%, p = 0.002) was the greatest contributor. In conclusion, regular water supplementation can improve the key enzyme activities and fungal metabolic function of sheep manure composting, and water replenishment once every 3.5 days had the best effect.

Effects of white-rot fungal pretreatment of corn straw return on greenhouse gas emissions from the North China Plain soil.

Straw-return with fungal treatment is a potential method for reducing soil greenhouse gas emissions through carbon (C) sequestration and N2O mitigation. However, there is little information on the effects of different fungal treatments of crop straw return on soil CO2 and N2O emissions. To explore to what extent decomposed corn straw and its components controls soil CO2 and N2O emissions, we set up three sequential incubation experiments using soil collected from the North China Plain, an intensive agricultural area. Interactions between the different C contents of corn straw (CS), CS pretreated with Irpex lacteus (ICS), CS pretreated with Phanerochaete chrysosporium (PCS) and different NO3--N concentrations on the effect of soil CO2 and N2O emissions were conducted, and the kinetics of CO2 and N2O as influenced by changes in soil biochemical factors were analyzed. The effects of different lignocellulose components (lignin, cellulose, and xylan) on soil CO2 and N2O emissions were further studied. The results showed that straw pretreatment did not affect CO2 emissions. Both CO2 and N2O emissions increased when the C and N contents increased. However, applying PCS to 70% water-filled pore space soil effectively decreased the soil N2O emissions, by 41.8%-76.3% compared with adding the same level of CS. Moreover, extracellular enzyme activities related to C and N cycling were triggered, and the nosZI and nosZII abundances were significantly stimulated by the PCS application. These effects are closely related to the initial soluble C content of this treatment. Furthermore, adding xylan can significantly reduce N2O emissions. Overall, our data suggest that the environmentally beneficial effects of returning straw can be greatly enhanced by applying the straw-degrading white-rot fungi of P. chrysosporium in the North China Plain soil. Future studies are needed in the field to upscale this technology.