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Abstract Fruit acidity significantly influences fruit flavor, but the specific impact of soil pH on fruit acidity remains unclear. This study investigated the effects of various soil pH levels on fruit acidity and citric acid (CA) metabolism in lemon (Citrus limon L.). High soil pH (pH 8) decreased total soluble solids concentrations in lemon fruits, while low soil pH (pH 4) increased titratable acid and CA concentrations. Although low soil pH reduced the synthesis of CA due to the decreased citrate synthase and phosphoenolpyruvate carboxylase activities, the elevated fruit acidity under low soil pH conditions is not directly related to CA synthesis. Instead, low soil pH was found to suppress the activity of cytosolic aconitase (Cyt-ACO), an iron-dependent enzyme, indicating a potential role for CA degradation inhibition in low soil pH-induced CA accumulation. Furthermore, low soil pH significantly reduced cytosolic iron (Cyt-Fe) concentration, which was positively correlated with Cyt-ACO activity. In conclusion, low soil pH contributes to increasing fruit acidity in lemon, partially by inhibiting CA degradation due to the reduced Cyt-Fe concentrations. Our work unravels the influence of soil pH on CA accumulation and provides important clues for modulating CA levels through microelement fertilization in citrus.

This study proposed an improved bio-carbonation of reactive magnesia cement (RMC) method for dredged sludge stabilization using the urea pre-hydrolysis strategy. Based on unconfined compression strength (UCS), pickling-drainage, and scanning electron microscopy (SEM) tests, the effects of pre-hydrolysis duration (T), urease activity (UA) and curing age (CA) on the mechanical properties and microstructural characteristics of bio-carbonized samples were systematically investigated and analyzed. The results demonstrated that the proposed method could significantly enhance urea hydrolysis and RMC bio-carbonation to achieve efficient stabilization of dredged sludge with 80% high water content. A significant strength increment of up to about 1063.36 kPa was obtained for the bio-carbonized samples after just 7 d of curing, which was 2.64 times higher than that of the 28-day cured ordinary Portland cement-reinforced samples. Both elevated T and UA could notably increase urea utilization ratio and carbonate ion yield, but the resulting surge in supersaturation also affected the precipitation patterns of hydrated magnesia carbonates (HMCs), which weakened the cementation effect of HMCs on soil particles and further inhibited strength enhancement of bio-carbonized samples. The optimum formula was determined to be the case of T = 24 h and UA = 10 U/mL for dredged sludge stabilization. A 7-day CA was enough for bio-carbonized samples to obtain stable strength, albeit slightly affected by UA. The benefits of high efficiency and water stability presented the potential of this method in achieving dredged sludge stabilization and resource utilization. This investigation provides informative ideas and valuable insights on implementing advanced bio-geotechnical techniques to achieve efficient stabilization of soft soil, such as dredged sludge.

Abstract Background Deciphering the assembly rules of microbial communities is vital for a mechanistic understanding of the general principles driving microbiome structures and functions. In this study, a null modeling-based framework was implemented to infer the assembly rules of bacterial community in oat silages harvested in southern China starting from the grain-filling stage through to full ripening. Results Most silages displayed “inferior” or “very inferior” fermentation quality. The fermentation qualities of silages tended to further decrease with the delay of harvest. Lactobacillus, Pediococcus, unclassified_f_Enterobacteriaceae, and Hafnia–Obesumbacterium constituted the predominated genera in silages. Delaying harvest increased the proportions of Hafnia–Obesumbacterium. Null model analysis revealed that stochastic processes were the primary contributor to the assembly of rare subcommunity during silage fermentation. The succession of abundant subcommunity was controlled both by stochastic and deterministic processes. Deterministic processes, more specifically, heterogeneous selection, were more prominent in the assembly of abundant bacteria in silages with the delay of harvest. Linear regression analysis indicated the important roles of DM, WSC and pH in the assembly of abundant subcommunity. Conclusion This study, from the ecological perspectives, revealed the ecological processes controlling the bacterial community assembly in silage, providing new insights into the mechanisms underlying the construction of silage bacterial community. Graphical Abstract

Abstract This study combined applied PICRUSt2 and BugBase tools to evaluate the effects of these two strains on the fermentation characteristics, microbial community, potential microbial risk and hygienic quality of whole-crop corn (WCC) silage. Fresh WCC harvested at the dough stage was ensiled with distilled water (CON), Leuconostoc lactis (LS) and Weissella confusa (WA) for 2, 4, 8, 15 and 30 days. After ensiling, all WCC silages presented desirable fermentation with high lactic acid and Lactobacillus proportions, low pH and ammonia nitrogen levels and absent butyric acid. Ensiling decreased the complexity of bacterial co-occurrence networks, and the Lc. lactis and W. confusa inoculation further decreased the complexity. The inoculation of W. confusa suppressed the most pathogenic pathways and related modules associated with zoonosis. In bacterial phenotype predicted analysis, although CON had lower proportions of ‘Potentially pathogenic’ than fresh material, this undesirable phenotype declined to negligible levels via LS and WA inoculation. Even for well-fermented WCC silages, the risk of pathogens remained after 30 days of ensiling. Therefore, WA could be developed as a promising fast start-up inoculant for reducing pathogenic contamination and improving hygienic quality of silage. Graphical Abstract

The objective was to evaluate the effectiveness of lactic acid bacteria (LAB) inoculants and chemical additives on the fermentation quality, aerobic stability and in vitro gas production kinetics and digestibility of total mixed ration (TMR) silage. Total mixed ration (568 g/kg dry matter (DM)) was ensiled with six experimental treatments: (1) no additives (control); (2) Lactobacillus buchneri (LB; applied at 1 × 106 cfu/g fresh weight (FW)); (3) Lactobacillus casei (LC; applied at 1 × 106 cfu/g FW); (4) calcium propionate (CAP; applied at 0.5% FW); (5) sodium diacetate (SD; applied at 0.5% FW); (6) potassium sorbate (PS; applied at 0.1% FW). All silos (18 L) were opened for fermentation quality, in vitro gas production kinetics and digestibility analysis after 90 days of ensiling, and then subjected to aerobic stability test for 14 days. All the TMR silage was well-preserved with low pH (4.36 ∼ 4.66) and acceptable levels of butyric acid (1.02 ∼ 2.51 g/kg DM) and ammonia nitrogen (86.3 ∼ 107 g/kg total nitrogen). All the groups were steady during 14 days of aerobic exposure, while SD group was more stable with lower (p

The current work aimed to evaluate the effect of ensiling density on the fermentation quality and aerobic stability of Pennisetum giganteum silages. The silage was ensiled in laboratory silos (1 L), and three treatments were designed according to different ensiling densities: (1) low density (LD, 750 kg/m3); (2) medium density (MD, 900 kg/m3); and (3) high density (HD, 1050 kg/m3). During ensiling, the silage was sampled for a fermentation quality analysis. All silages were well preserved, as indicated by the high lactic acid (LA) content and low pH (3-N) content (68.2 g/kg total nitrogen, TN) among all silages after 45 days of ensiling. The aerobic stability of HD, MD, and LD persisted for 51 h, 54 h, and 48 h, respectively. The NH3-N contents of HD and MD were below 80 g/kg TN during aerobic exposure. These results show that the increase in ensiling density improved the fermentation quality and aerobic stability of Pennisetum giganteum silages.

Abstract Organic photovoltaics have attracted worldwide interest due to their unique advantages in developing low-cost, lightweight, and flexible power sources. Functional molecular design and synthesis have been put forward to accelerate the discovery of ideal organic semiconductors. However, it is extremely expensive to conduct experimental screening of the wide organic compound space. Here we develop a framework by combining a deep learning model (graph neural network) and an ensemble learning model (Light Gradient Boosting Machine), which enables rapid and accurate screening of organic photovoltaic molecules. This framework establishes the relationship between molecular structure, molecular properties, and device efficiency. Our framework evaluates the chemical structure of the organic photovoltaic molecules directly and accurately. Since it does not involve density functional theory calculations, it makes fast predictions. The reliability of our framework is verified with data from previous reports and our newly synthesized organic molecules. Our work provides an efficient method for developing new organic optoelectronic materials.

Abstract Background The purpose of this research is to study the contribution of epiphytic microbiota in fresh oat (OT), Italian ryegrass (IR) and whole-crop maize (MZ) to silage fermentation products and bacterial community structure of MZ. After γ-ray irradiation, the sterile MZ was treated via microbiota transplantation method: (1) sterile deionized water (STMZ); (2) microbiota epiphytic on MZ (MZMZ); (3) microbiota epiphytic on OT (MZOT); (4) microbiota epiphytic on IR (MZIR). Triplicate silos of each treatment were tested after 1, 3, 7, 15, 30 and 60 days of ensiling. Results MZMZ had higher (P

The inefficiency observed in investment within state-owned enterprises presents a significant practical challenge that can affect the sustainable development of China's economy. To address this issue, this study comprehensively explores the intricate mechanisms underlying the governance implications of mixed ownership on the investment efficiency of listed companies. Drawing on unbalanced panel data encompassing Shanghai and Shenzhen Stock Exchange A-share listed companies in China spanning the period from 2008 to 2022, this study employs a fixed-effects model to unveil the nuanced ways in which mixed ownership influences investment efficiency through the lens of agency costs. This study transcends the boundaries of traditional agency conflicts between managers and shareholders. It delves deeper, illuminating the diverse effects of agency conflicts between significant controlling shareholders and minority shareholders. The results revealed a noteworthy positive correlation between mixed ownership and investment efficiency, and verified the intermediary role of agency costs between mixed ownership and investment efficiency, which is an important result of our research. Heterogeneity analysis indicates that the relationship between the two can be affected by external events, such as during the COVID-19 pandemic, investment efficiency is not the most concerned issue for enterprises. The findings have practical implications for practitioners and policymakers, as they offer avenues for optimizing investment strategies and fostering efficient and effective corporate governance practices.

Abstract Background Italian ryegrass is a temperate climate crop, which is widely cultivated in the winter fallow paddy fields of subtropical China. The utilization efficiency of Italian ryegrass depends greatly on its growth stage at harvest. Previous studies have reported the optimum stage for harvesting various forage to balance their quality and quantity. However, when considering the practice condition, such as rainy or unavailability of harvest equipment, the harvest stage of forage cannot always be implemented according to the production schedules. Thus, to characterize the effect of growth stage on the silage fermentation profile, bacterial community construction and metabolisms of carbohydrates and amino acids, Italian ryegrass were naturally ensiled at the filling stage (FSN) and the dough stage (DSN), respectively. After ensiling for 1, 3, 7, 15, 30, and 60 days, triplicate silos were opened for sampling. Results The growth of Italian ryegrass increased the pH, dry matter, neutral, and acid detergent fiber contents, but decreased buffering capacity, crude protein and water-soluble carbohydrates contents, and the epiphytic microbiota of Italian ryegrass harvested at the filling stage was simpler than that harvested at the dough stage. During ensiling, FSN had lower pH and higher organic acid content than DSN. The bacterial succession rate in FSN was also faster than DSN, which showed that Lactobacillus becomes the dominant genus in the early stage of ensiling. The predicted metabolisms revealed that carbohydrate and amino acid metabolisms were the two main metabolisms in silage fermentation. When compared with epiphytic microbiota, ensiling enhanced carbohydrate metabolism and diminished amino acid metabolism. The difference of these two metabolisms between FSN and DSN was obvious at the early stage of ensiling. Conclusions Growth stage affected the chemical and microbial composition of Italian ryegrass, so as to the fermentation profile, bacterial community and its metabolisms intensity. Italian ryegrass harvested at the filling stage was prone to ferment. The complexity of epiphytic microbiota made Italian ryegrass harvested at the dough stage more difficult to ferment. Graphical Abstract

Abstract Background This study evaluated the effects of growth stage and storage time on fermentation characteristics, bacterial communities and their functionality in alfalfa (Medicago sativa L.) silage. Alfalfa was harvested at initial flowering (10–20% bloom, AL1) and full flowering (> 80% bloom, AL2) stages, respectively. The harvested alfalfa was ensiled in 15 L plastic silos. Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of ensiling, respectively. Fermentation products were analyzed on each sampling day. The bacterial communities and their functional potential after 3 and 60 days were analyzed by high-throughput sequencing technique and PICRUSt2 method. Results AL2 had better fermentation quality than AL1 with lower pH, ammonia nitrogen and butyric acid concentrations and higher lactic acid concentrations on day 60. AL2 had higher abundances of Weissella and Lactobacillus after 3 days, and lower abundances of Enterobacter and Enterobacteriaceae on day 60 compared to AL1. In metabolic pathway analysis, ensiling promoted the carbohydrate and amino acid metabolism, and inhibited the signal transduction and membrane transport. In enzyme analysis, AL2 had lower abundances of nitrite reductase (NADH) and ornithine decarboxylase than AL1 on day 60. In phenotype analysis, AL2 had higher proportions of facultatively anaerobic and lower proportions of anaerobic, potentially pathogenic and gram negative than AL1 on day 60. Conclusions High throughput sequencing technique combined with PICRUSt2 can be successfully used to describe the changes of bacterial communities and their functionality in silage. This approach can improve our understanding of the silage microbiology to further regulate the fermentation products. Graphical Abstract

This study was conducted to evaluate the effects of different additives on the fermentation quality and bacterial community of silage prepared from fresh-cut whole-plant quinoa (WPQ). Fresh-cut WPQ without wilting process was directly ensiled with one of the following additives: (i) untreated (control); (ii) fibrolytic enzyme (E); (iii) molasses (M); (iv) LAB inoculant (L); (v) a combination of fibrolytic enzyme and LAB inoculant (EL); and (vi) a combination of molasses and LAB inoculant (ML). The fermentation quality and bacterial community after 60 days of ensiling were analysed. The results showed that control silage was dominated by enterobacteria (>50%), with high pH value (5.25), acetic acid (77.9 g/kg DM) and NH3–N production (261 g/kg TN). Adding E, EL and ML shifted the fermentation pattern towards increased intensity of lactic acid production, as indicated by increased lactic acid content (>62.4 g/kg DM), lactic/acetic acid ratio (>1.12) and relative abundance of Lactobacillus (>62.0%), and decreased pH value (

Objective The observation that temperate C3 and tropical C4 forage silages easily produce large amounts of ethanol or acetic acid has puzzled researchers for many years. Hence, this study aimed to assess the effects of epiphytic microbiota from C3 forages (Italian ryegrass and oat) on fermentative products and bacterial community structure in C4 forage (sorghum) silage. Methods Through microbiota transplantation and γ-ray irradiation sterilization, the irradiated sorghum was treated: i) sterile distilled water (STSG); ii) epiphytic microbiota from sorghum (SGSG); iii) epiphytic microbiota from Italian ryegrass (SGIR); iv) epiphytic microbiota from oat (SGOT). Results After 60 days, all the treated groups had high lactic acid (>63.0 g/kg dry matter [DM]) contents and low pH values (

Objective This study aimed to investigate the fermentation profiles, bacterial community and predicted metabolic characteristics of Sudangrass (Sorghum sudanense Stapf.) during ensiling. Methods First-cutting Sudangrass was harvested at the vegetative stage and ensiled in laboratory-scale silos (1 L capacity). Triplicate silos were sampled after 1, 3, 7, 15, 30, and 60 days of ensiling, respectively. The bacterial communities on day 3 and 60 were assessed through high-throughput sequencing technology, and 16S rRNA-gene predicted functional profiles were analyzed according to the Kyoto encyclopedia of genes and genomes using Tax4Fun. Results The Sudangrass silages showed good fermentation quality, indicated by higher lactic acid contents, and lower pH, butyric acid and ammonia nitrogen contents. The dominant genus Lactococcus on day 3 was replaced by Lactobacillus on day 60. The metabolism of amino acid, energy, cofactors and vitamins was restricted, and metabolism of nucleotide and carbohydrate was promoted after ensiling. The 1-phosphofructokinase and pyruvate kinase of bacterial community seemed to play important roles in stimulating the lactic acid fermentation, and the promotion of arginine deiminase could help lactic acid bacteria to tolerate the acidic environment. Conclusion High-throughput sequencing technology combined with 16S rRNA gene-predicted functional analyses revealed the differences during the early and late stages of Sudangrass ensiling not only for distinct bacterial community but also for specific functional metabolites. The results could provide a comprehensive insight into bacterial community and metabolic characteristics to further improve the silage quality.

ABSTRACT Forage epiphytic microbiota exhibits pronounced changes in composition and function throughout the day. However, the effects of these changes on silage fermentation are rarely explored. Here, we transplanted the epiphytic microbiota of sorghum-sudangrass hybrid (SSG) harvested at 7:00 h (AM), 12:00 h (M), and 17:00 h (PM) to sterilized SSG to evaluate the effects of diurnal variation of epiphytic microbiota on fermentation characteristics. During fermentation, remarkable differences in bacterial community successions were observed between silages inoculated with AM and M microbiota. Compared to AM microbiota, M microbiota inoculation increased the proportions of Pantoea dispersa, Leuconostoc lactis, Enterobacter, and Klebsiella variicola, whereas it decreased the proportions of Weissella cibaria and Lactobacillus plantarum during fermentation. This led to the most rapid pH declines and organic acid production in AM silage and the slowest in M silage. Both M and PM microbiota affected the bacterial cooccurrence patterns, indicated by decreased complexity and stability in the community structures of M and PM silages compared to that of AM silage. The predicted functions indicated that some key carbohydrate metabolism pathways related to lactic acid synthesis were downregulated, while some competing pathways (ascorbate and aldarate metabolism and C5-branched dibasic acid metabolism) were upregulated in M silage compared to AM silage after 3 days of fermentation. Correlation analysis revealed positive correlations between competing pathways and enterobacterial species. The current study highlights the importance of diurnal variation of epiphytic microbiota in affecting the silage bacterial community, potentially providing an effective strategy to improve silage quality by optimizing harvest time. IMPORTANCE Ensiling is a way to preserve wet biomass for animal and bioenergy production worldwide. The fermentation quality of silage is largely dependent on the epiphytic microbiota of the material. Plant epiphytic microbiota exhibit diurnal changes in composition and function. However, the effects of these changes on silage fermentation are rarely explored. The results presented here demonstrated that diurnal variation of epiphytic microbiota could affect the fermentation characteristics and bacterial community during SSG fermentation. Marked bacterial community differences were observed between AM and M silages during the initial 3 days of fermentation. The dominance rate of Lactobacillus plantarum was highest in AM silage, whereas enterobacterial species were more abundant in M silage. The predicted function revealed downregulated lactic acid synthesis pathways and upregulated competing pathways in M silage compared to those in AM silage. This study provides clues for technological-parameter optimization of the fermentation process by the selection of harvest time.

Abstract Introduction The Canadian Cannabis Act came into effect on October 17, 2018, which allowed Canadian adults to consume cannabis for non-medical purposes (Government of Canada, Cannabis regulations (SOR/2018-144). Cannabis Act, (2018a); Parliament of Canada, C-45: an Act respecting cannabis and to amend the Controlled Drugs and Substances Act, the Criminal Code and other Acts, 2018). With this major policy change, it is unknown how the attitude of the public changed and how information on cannabis changed. Social media platforms, including Twitter, are significant venues for studying emerging patterns in social issues such as cannabis legalization. This study aimed to examine sentiments, themes and contents of cannabis-related tweets by suppliers (both licensed and unlicensed) and general tweets in Canada. To our knowledge, this is the first analysis that mixes sentiment analysis and thematic analysis of Canadians’ cannabis-related Twitter data. Method A sample of Canadian cannabis-related tweets was collected from January 2018 to August 2020 through the Twitter Application Programming Interface (API). Using a standard access token and the Twitter Standard Search API, tweets were extracted based on Twitter handles to capture the content of both licensed and unlicensed cannabis retailers in Canada, as well as relevant cannabis-related keywords to capture public content. We conducted sentiment and positive polarity analyses, and content analysis to identify attitudes and themes around cannabis use in Canada. Results This study gathered and analyzed a total of 44,970 tweets in the sentiment analysis and a total of 1035 tweets in the thematic analysis. Descriptive analysis showed that monthly tweets peaked prior to legalization in October 2018 and again during the initial wave of the COVID-19 pandemic in February and March 2020. The data showed an overall positive sentiment polarity with a high of + 0.24 in April 2019 and a low of + 0.14 in March 2020. Thematic analysis revealed the themes: (i) education/information, (ii) uses of cannabis, (iii) cannabis products including packing, quality, price, types, and sources, (iv) cannabis policies including regulations and public safety, (v) access, (vi) social issues include gender and stigma, and (vii) COVID-19 impact. Conclusion This study combined the power of big data collection and analysis with manual coding and analysis methods to extract rich content from large data using social media communications on issues related to cannabis in Canada. The findings of this study may inform policies on advertising cannabis products and highlighted some patterns related to education, access, and safety that deserve further investigation.

Abstract Background In early lactation, bovine mammary epithelial cells undergo serious metabolic challenges and oxidative stress both of which could be alleviated by activation of autophagy. Nuclear factor erythroid 2 related factor 2 (NFE2L2), a master regulator of cellular redox homeostasis, plays an important role in the regulation of autophagy and oxidative stress. Thus, the objective of this study was to investigate the role of NFE2L2-mediated autophagy on oxidative stress of bovine mammary epithelial cells in response to exogenous free fatty acids (FFA). Results Exogenous FFA induced linear and quadratic decreases in activities of glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD), and increases in the contents of reactive oxygen species (ROS) and malondialdehyde (MDA). Protein abundance of LC3-phosphatidylethanolamine conjugate (LC3-II) and the number of autophagosomes and autolysosomes decreased in a dose-dependent manner, while protein abundance of p62 increased in cells challenged with FFA. Activation of autophagy via pre-treatment with Rap attenuated the FFA-induced ROS accumulation. Importantly, FFA inhibited protein abundance of NFE2L2 and the translocation of NFE2L2 into the nucleus. Knockdown of NFE2L2 by siRNA decreased protein abundance of LC3-II, while it increased protein abundance of p62. Furthermore, sulforaphane (SFN) pre-treatment attenuated the FFA-induced oxidative stress by activating NFE2L2-mediated autophagy. Conclusions The data suggested that NFE2L2-mediated autophagy is an important antioxidant mechanism in bovine mammary epithelial cells experiencing increased FFA loads.

ABSTRACT To characterize the effects of time of day for harvest on the fermentation parameters, bacterial community, and metabolic characteristics of sorghum-sudangrass hybrid (SSG) silage, SSG (vegetative stage) harvested at 7:00 (AM), 12:00 (M), and 17:00 (PM) on three sunny days were ensiled for 1, 3, 7, 14, 30, and 60 days. Compared to AM silage, M and PM silages were characterized by delayed fermentation, unnormal lower final pH, and lower acetic acid production. In addition, PM silage contained higher residual water-soluble carbohydrates than other silages. After 60 days of ensiling, AM silage was dominated by Lactobacillus, whereas the bacterial communities of M and PM silages were complex and mainly composed of bacteria such as Delftia, Methylobacterium-Methylorubrum, Enhydrobacter, Acinetobacter, and Bacillus. The harvest time affected a wide range of metabolic pathways including “Metabolism” and “Cellular Processes” and “Organismal Systems” in SSG silage. Particularly, at the late stage of ensiling M silage exhibited highest relative abundances of amino acid metabolisms including “glycine, serine, and threonine metabolism,” “phenylalanine metabolism,” and lowest relative abundances of “lysine biosynthesis.” These results suggest that the time of day for harvest could affect the fermentation parameters, bacterial community, and metabolic characteristics of SSG silage. Better SSG silage characteristics could be achieved through morning harvest. IMPORTANCE Ensiling is a common way for preserving green forages worldwide. Silage fermentation quality can vary greatly depending on the chemical and microbial characteristics of forage crop being ensiled. It is well documented that forages exhibit considerable variations in chemical composition and epiphytic microbiota during daylight. However, the effects of the time of day for harvest on silage fermentation is less investigated. Our results demonstrate that the time of day for harvest could affect the fermentation parameters, bacterial community, and metabolic characteristics of SSG hybrid silage. Harvesting SSG late in the day delayed fermentation process, lowered acetic acid production and final pH, and increased the residual water-soluble carbohydrates content in silage. Moreover, the delayed harvest time increased the relative abundances of bacteria such as Delftia, Methylobacterium-Methylorubrum, Acinetobacter, Enhydrobacter, and Bacillus, and amino acid metabolisms at the late stage of SSG ensiling. This study highlights the importance of diurnal changes in forage to fermentation characteristics, providing a strategy to improve silage quality through optimizing the harvest time.

Cows with ketosis display severe metabolic stress and immune dysfunction which renders them more susceptible to infections. Monocytes, one of the major subtypes of white blood cells, play an important role in innate immune defense against infections. Thus, the aim of this study was to investigate alterations in immune function, reactive oxygen species (ROS) production and activity of the NLR family pyrin domain containing 3 (NLRP3) inflammasome pathway in monocytes (CD14+) of cows with clinical ketosis (CK). Twelve healthy multiparous Holstein cows [blood β-hydroxybutyrate (BHB) concentration < 1.2 mM] and 12 cows with CK (BHB > 3.0 mM) at 3 to 14 days in milk were used for blood sample collection. To determine effects of BHB on phagocytosis, ROS and protein abundance of the NLRP3 inflammasome pathway in vitro, monocytes isolated from healthy cows were treated with 3.0 mM BHB for 0, 6, 12 or 24 h. Dry matter intake (22.7 vs. 19.0 kg) was lower in cows with CK. Serum concentrations of fatty acids (0.30 vs. 0.88 mM) and BHB (0.52 vs. 3.78 mM) were greater in cows with CK, whereas concentration of glucose was lower (4.09 vs. 2.23 mM). The adhesion, migration and phagocytosis of monocytes were lower in cows with CK, but apoptosis and ROS content were greater. Protein abundance of NLRP3, cysteinyl aspartate specific proteinase 1 (caspase 1) and interleukin-1B p17 (IL1B p17) were greater in monocytes of cows with CK, while abundance of NADPH oxidase isoform 2 (NOX2) was lower. Compared with 0 h BHB, ROS content and apoptosis were greater in the monocytes challenged for 6, 12 or 24 h BHB. Compared with 0 h BHB, protein abundance of NLRP3, caspase 1, IL1B p17 and concentration of IL1B in medium were greater in the monocytes challenged for 6, 12 or 24 h BHB. However, compared with 0 h BHB, protein abundance of NOX2 and phagocytosis of monocytes were lower in the monocytes challenged for 6, 12 or 24 h BHB. Overall, the data suggested that exogenous BHB activated the ROS-NLRP3 pathway, which might be partly responsible for immune dysfunction of dairy cows with CK.

Objective The study aimed to evaluate the effect of storage time and formic acid (FA) on fermentation characteristics, epiphytic microflora, carbohydrate components and in vitro digestibility of rice straw silage. Methods Fresh rice straw was ensiled with four levels of FA (0%, 0.2%, 0.4%, and 0.6% of fresh weight) for 3, 6, 9, 15, 30, and 60 d. At each time point, the silos were opened and sampled for chemical and microbial analyses. Meanwhile, the fresh and 60-d ensiled rice straw were further subjected to in vitro analyses. Results The results showed that 0.2% and 0.6% FA both produced well-preserved silages with low pH value and undetected butyric acid, whereas it was converse for 0.4% FA. The populations of enterobacteria, yeasts, moulds and aerobic bacteria were suppressed by 0.2% and 0.6% FA, resulting in lower dry matter loss, ammonia nitrogen and ethanol content (p0.05). Conclusion The 0.2% FA application level promoted lactic acid fermentation while 0.6% FA restricted all microbial fermentation of rice straw silages. Rice straw ensiled with 0.2% FA or 0.6% FA improved its nutrient preservation without affecting digestion, with the 0.6% FA level best.

In this study, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed to investigate the polymorphism of GHR gene and the correlations of polymorphic sites with growth traits. The results showed that GHR detected CC, CD and DD genotypes in the five sheep populations. GHR gene had the highest DD genotype frequencies in Yuxi fatty-tailed sheep, Dorper sheep, Henan large-tailed han sheep, Hu sheep and small-tailed han sheep, being 0.889, 0.784, 0.785, 0.529 and 0.750, respectively. The correlation analysis indicated that GHR gene had significant correlations with chest circumference and abdomen circumference of sheep

Abstract Complex networks in the real world are often with heterogeneous degree distributions. The structure and function of nodes can vary significantly, with vital nodes playing a crucial role in information spread and other spreading phenomena. Identifying and taking action on vital nodes enables change to the network’s structure and function more efficiently. Previous work either redefines metrics used to measure the nodes’ importance or focuses on developing algorithms to efficiently find vital nodes. These approaches typically rely on global knowledge of the network and assume that the structure of the network does not change over time, both of which are difficult to achieve in the real world. In this paper, we propose a localized strategy that can find vital nodes without global knowledge of the network. Our joint nomination (JN) strategy selects a random set of nodes along with a set of nodes connected to those nodes, and together they nominate the vital node set. Experiments are conducted on 12 network datasets that include synthetic and real-world networks, and undirected and directed networks. Results show that average degree of the identified node set is about 3–8 times higher than that of the full node set, and higher-degree nodes take larger proportions in the degree distribution of the identified vital node set. Removal of vital nodes increases the average shortest path length by 20–70% over the original network, or about 8–15% longer than the other decentralized strategies. Immunization based on JN is more efficient than other strategies, consuming around 12–40% less immunization resources to raise the epidemic threshold to $$\tau \sim 0.1$$ τ ∼ 0.1 . Susceptible-infected-recovered simulations on networks with 30% vital nodes removed using JN delays the arrival time of infection peak significantly and reduce the total infection scale to 15%. The proposed strategy can effectively identify vital nodes using only local information and is feasible to implement in the real world to cope with time-critical scenarios such as the sudden outbreak of COVID-19.

Abstract Background Bovine mammary epithelial cells after calving undergo serious metabolic challenges and oxidative stress both of which could compromise autophagy. Transcription factor EB (TFEB)-mediated autophagy is an important cytoprotective mechanism against oxidative stress. However, effects of TFEB-mediated autophagy on the oxidative stress of bovine mammary epithelial cells remain unknown. Therefore, the main aim of the study was to investigate the role of TFEB-mediated autophagy in bovine mammary epithelial cells experiencing oxidative stress. Results H2O2 challenge of the bovine mammary epithelial cell MAC-T increased protein abundance of LC3-II, increased number of autophagosomes and autolysosomes while decreased protein abundance of p62. Inhibition of autophagy via bafilomycin A1 aggravated H2O2-induced reactive oxygen species (ROS) accumulation and apoptosis in MAC-T cells. Furthermore, H2O2 treatment triggered the translocation of TFEB into the nucleus. Knockdown of TFEB by siRNA reversed the effect of H2O2 on protein abundance of LC3-II and p62 as well as the number of autophagosomes and autolysosomes. Overexpression of TFEB activated autophagy and attenuated H2O2-induced ROS accumulation. Furthermore, TFEB overexpression attenuated H2O2-induced apoptosis by downregulating the caspase apoptotic pathway. Conclusions Our results indicate that activation of TFEB mediated autophagy alleviates H2O2-induced oxidative damage by reducing ROS accumulation and inhibiting caspase-dependent apoptosis.

Objective To investigate the improvement in utilization efficiency of total mixed ration (TMR) on Tibetan plateau, TMR were ensiled with different additives. Methods A total of 150 experimental silos were prepared in a completely randomized design to evaluate the six treatments: i) control (without additive), ii) Lactobacillus buchneri (L. buchneri), iii) acetic acid, iv) propionic acid, v) 1,2-propanediol; and vi) 1-propanol. After 90 days of ensiling, silos were opened for fermentation quality and in vitro analysis, and then subjected to an aerobic stability test for 14 days. Results Treating with L. buchneri, acetic acid, 1,2-propanediol and 1-propanol decreased propionic acid contents and yeast number, whereas increased (p0.05) all in vitro parameters including gas production at 24 h (GP24), GP rate constant, potential GP, in vitro DM digestibility and in vitro neutral detergent fibre digestibility. All additives improved the aerobic stability of ensiled TMR to different extents. Specially, aerobic stability of the ensiled TMR were substantially improved by L. buchneri, acetic acid, 1,2-propanediol, and 1-propanol, indicated by stable pH and lactic acid content during the aerobic stability test. Conclusion L. buchneri, acetic acid, 1,2-propanediol, and 1-propanol had no adverse effect on in vitro digestibility, while ensiling TMR with the additives produced more acetic acid and ethanol, subsequently resulting in improvement of aerobic stability. There is a potential for some fermentation boosting additives to enhance aerobic stability of fermented TMR on Tibetan plateau.

This study aimed to investigate the impacts of three organic acid salts and two bacterial additives on the fermentation profile, in vitro digestibility and gas production, and aerobic stability of total mixed ration (TMR) silage prepared with 10% fresh weight (FW) of wet hulless barley distillers’ grains (WHDG), 35% FW of common vetch, 15% FW of whole-crop oat, 10% FW of hulless barley straw, and 30% FW of mixed concentrate. The total mixed ration (555 g kg−1 FW) was ensiled with six treatments: (1) no additives (control); (2) calcium propionate (CAP; 0.5% FW); (3) potassium sorbate (POS; 0.1% FW); (4) sodium diacetate (SDA; 0.5% FW); (5) Lactobacillus buchneri (LB; 1 × 106 cfu g−1 FW); and (6) Lactobacillus casei (LAC; 1 × 106 cfu g−1 FW). All silos (20 L) were opened for the fermentation profile and in vitro digestibility analysis after 95 days of fermentation, and then subjected to an aerobic stability experiment for 14 days. All the TMR silage was well preserved with a low pH (4.32~4.51), acceptable levels of butyric acid (1.34~1.56 g kg−1 dry matter), and ammonia nitrogen (69.1~87.1 g kg−1 total nitrogen). All the groups were steady during aerobic exposure, while the SDA treatment was more stable with lower populations of undesirable microorganisms on day 14. The CAP, POS, and SDA treatments evidently (p < 0.05) enhanced the in vitro digestibility of dry matter (54.8~57.5 vs. 48.4%) and neutral detergent fiber (48.4~51.6 vs. 41.1%) compared to the control. By comprehensive consideration, SDA is recommended as additive to enhance fermentation quality, aerobic stability, and in vitro digestibility of TMR silage containing 10% FW of WHDG.

This study aimed to evaluate the effect of substituting Pennisetum sinese with bamboo shoot shell (BSS) on the fermentation quality, chemical composition, in vitro digestibility and aerobic stability of ensiled total mixed ration (TMR). Four ensiled TMRs were designed on fresh matter basis: (i) 45% P. sinese (BSS0); (ii) 15% BSS + 30% P. sinese (BSS15); (iii) 25% BSS + 20% P. sinese (BSS25); (iv) 35% BSS + 10% P. sinese (BSS35). All silages were moderately fermented according to the V-score. Substituting P. sinese with BSS increased acetic acid content and decreased ethanol and ammonia nitrogen contents (p .05). The substitution of P. sinese with BSS had no adverse effect on the fermentation quality and in vitro digestibility while efficiently improving the aerobic stability of ensiled TMR. The BSS35 substitution level is recommended considering the maximum utilisation of BSS.HIGHLIGHTS Bamboo shoot shell (BSS) was explored for ensiled TMR. BSS had no effect on the silage digestibility. BSS improved aerobic stability.

An experiment was conducted to investigate the effects of citric acid residue (CAR) and lactic acid bacteria (LAB) on fermentation quality and aerobic stability of alfalfa silage. Alfalfa was treated as follows: (1) control without additive (CON); (2) LAB (L); (3) 12 g/kg CAR (C); (4) 12 g/kg CAR + LAB (CL) and fermented for 3, 6, 9, 15, 30 and 45 days. The residual silages ensiled for 45 days were evaluated for aerobic stability. The results showed that addition of CAR with or without LAB inoculant decreased pH, acetic acid, ammonia nitrogen , neutral detergent fibre, hemicellulose contents and Enterobacteriaceae counts (p

Most arthropod guts harbor diverse microbiota for symbiotic digestion. The European corn borer (ECB), Ostrinia nubilalis (Hübner), is a devastating pest that feeds the lignocellulose-rich tissues of maize plants. However, the potential role of ECB gut microbes in degrading maize cellulose remains largely unexplored. Here, we investigated the gut microbiota of ECB fed with different diets and their potential function in maize lignocellulose degradation. The diversity and composition of gut bacterial communities varied dramatically between the ECB larva fed with artificial diets (ECB-D) and maize plants (ECB-M). Draft genomes of the microbial consortia from ECB-D and ECB-M showed that the principal degraders of cellulose mainly belonged to Firmicutes or Proteobacteria and they were primarily found in the midgut. The cellulolytic microbial consortia contained genes encoding various carbohydrate-active enzymes (CAZyme). Furthermore, scanning electron microscopy revealed significant breakdown of lignocellulose in maize treated by the two microbial consortia for 9 days in vitro. Metabolomic analyses show that maize particles treated by two microbial consortia generate distinctive metabolomic profiles, with enrichment for different monosaccharides (i.e., Glucose, Rhamnofuranose, Isomaltose, and Cellobiose) and amino acids (i.e., Threonine, Histidine, and Lysine). The results indicated that the diet of the host impacted the composition and function of its gut microbiota and ECB exploited specific gut microbes to digest maize lignocellulose with distinctive products. Our study provides valuable microbiota resources for lignocellulose bioconversion.

This study aimed to investigate the effect of epiphytic microbiota from alfalfa and red clover on the fermentative products, bacterial community compositions, and their predicted functional characteristics in Italian ryegrass silage. By microbiota transplantation and γ-ray irradiation sterilization, the irradiated Italian ryegrass was treated as follows: (1) sterile distilled water (STIR); (2) epiphytic microbiota on Italian ryegrass (IRIR); (3) epiphytic microbiota on alfalfa (IRAL); and (4) epiphytic microbiota on red clover (IRRC). The irradiated Italian ryegrass was ensiled for 1, 3, 7, 15, 30, and 60 days. STIR had similar chemical components with fresh Italian ryegrass. IRAL had higher lactic acid concentrations [64.0 g/kg dry matter (DM)] than IRIR (22.3 g/kg DM) and IRRC (49.4 g/kg DM) on day 3. IRRC had the lowest lactic acid concentrations (59.7 g/kg DM) and the highest pH (4.64), acetic acid (60.4 g/kg DM), ethanol (20.4 g/kg DM), and ammonia nitrogen (82.6 g/kg DM) concentrations and Enterobacteriaceae [9.51 log10 cfu/g fresh weight (FW)] populations among treatments on day 60. On days 3 and 60, Lactobacillus was dominant in both IRIR (42.2%; 72.7%) and IRAL (29.7%; 91.6%), while Hafnia-Obesumbacterium was predominant in IRRC (85.2%; 48.9%). IRIR and IRAL had lower abundances of “Membrane transport” than IRRC on day 3. IRIR and IRAL had lower abundances of phosphotransacetylase and putative ATP-binding cassette transporter and higher abundances of arginine deiminase on day 3. IRAL had the highest abundance of fructokinase on day 3. Overall, inoculating epiphytic microbiota from different legume forages changed the fermentative products, bacterial community compositions, and their predicted functional characteristics in Italian ryegrass silage. The microbial factors that result in the differences in fermentative profiles between legume forage and grass were revealed. Knowledge regarding the effect of epiphytic microbiota could provide more insights into the improvement of silage quality.

Objective To explore feed resources capable of replacing regular poor-quality fodder, this study was conducted to evaluate the effects of additives on the fermentation quality, in vitro digestibility and aerobic stability of mulberry leaves silage. Methods The mulberry leaves were ensiled either untreated (control) or treated with 1×106 cfu/g fresh matter Lactobacillus plantarum (L), 1% glucose (G), 3% molasses (M), a combination of 1% glucose and Lactobacillus plantarum (L+G), and a combination of 3% molasses and Lactobacillus plantarum (L+M). The fermentation quality and chemical composition were analyzed after 7, 14, 30, and 60 d, respectively. The 60-d silages were subjected to an aerobic stability test and fermented with buffered rumen fluid to measure the digestibility. Results Inoculating lactic acid bacteria (LAB) resulted in more rapid increase in lactic acid concentrations and decline in pH of mulberry leaves silage as compared control. Higher acetic acid and lower ethanol and ammonia nitrogen concentrations (p

This study aimed to investigate the effect of fibrolytic enzymes, cellulolytic fungi and lactic acid bacteria on the fermentation quality, structural carbohydrate composition and in vitro digestibility of rice straw silage. This experiment followed a completely randomised block design; four treatments were designed: (1) distilled water (control, CON); (2) fibrolytic enzymes (2.0 g/kg fresh weight (FW), E); (3) Trichoderma reesei (4400 U/kg FW, F); (4) Enterococcus faecium Y83 (1 × 106 cfu/g FW, Y83). All additives were diluted with distilled water and sprayed onto the rice straw (20 mL/kg FW). The rice straw was placed into a laboratory silo (10 L) after uniformly mixing and stored at ambient temperature (17–22 °C) ensiling for 3, 7, 14, 30 and 60 days. The fermentation quality in treated silages was improved compared to the CON, as indicated by lower pH, propionic acid, acetic acid and ammonia nitrogen (NH3-N) contents. Furthermore, Y83 had the lowest (p < 0.05) pH and highest (p < 0.05) lactic acid content after 60 days of ensiling. Y83 significantly (p < 0.05) decreased the neutral detergent fibre content compared with CON, E and F. In addition, E and Y83 had significantly (p < 0.05) higher in vitro dry matter digestibility and in vitro neutral detergent fibre digestibility than CON and F. Overall, Y83 can be used as a promising inoculant for improving the fermentation quality of rice straw silage.

Chlorine (Cl) is indispensable for the growth of plants. While rarely systematic reports are available for the effect of Cl-containing fertilizers on citrus production. This study aimed to investigate the impacts of various Cl-containing fertilizers on the nutrients in the leaves, the yield and quality of sweet orange, and the Cl migration in the plant–soil–leaching system. A 5-year field experiment (2016–2020) with five Cl treatments (0, 75, 150, 450, and 900 kg ha−1), and soil core lysimeter test with five Cl levels (0, 150, 225, 300, and 450 kg ha−1) were carried out. The results showed that 77.0% of Cl leached into above 60 cm deeper soil layer, with calcium as the main accompanying ions, resulting in less Cl being absorbed by the citrus plants. The content of Cl in the leaves and soil was enhanced by the increasing input of Cl-containing fertilizer, without yearly increased characteristics, under a mean annual rainfall of 1,474 mm. Chlorine significantly increased the yield (13.24–37.8 9%), fruit weight, and vitamin C (Vc), in addition to enhancing the flavor and the juice yield of sweet orange via improving the absorption of N and K. Moreover, the long-term application of potassium sulfate has elevated the accumulation of sulfur in the soil and in leaves; it is becoming a potential risk factor for citrus production. Taken together, the application of Cl-containing fertilizer in sweet orange is feasible, and trace absorbance of Cl could improve the yield and fruit quality of sweet orange.

The effects of sorbic acid, ethanol, previously fermented juice and their combined additives on fermentation quality and residual water-soluble carbohydrates (WSC) of napiergrass (Pennisetum purpureum) were herein investigated. There were two experiments in total. The treatments of experiment 1 were as follows: control (no addition); sorbic acid at 0.1%, S; molasses at 1.0%, M; previously fermented juice at 5 mL/kg, P; SP; SM; SPM. The treatments of experiment 2 were as follows: control (no addition); ethanol at 1.5%, E; molasses at 1.0%, M; previously fermented juice at 5 mL/kg, P; EM; EP; EPM. The laboratory silos (10 L) were kept at room temperature (~25 °C), and opened after 120 days of ensiling and the chemical compositions of silage were analyzed. Sorbic acid addition significantly (p < 0.05) decreased the silage pH and increased the residual WSC and lactic acid (LA) contents as compared with control. Ethanol addition decreased pH, acetic acid (AA) and volatile fatty acids (VFAs) contents, and increased the ratio of LA/AA, LA and residual WSC contents. Molasses addition increased ratios of LA/AA and enhanced WSC content used by lactic acid bacteria (LAB). The PFJ addition increased AA and ammonia nitrogen/total nitrogen (AN/TN) and decreased the LA contents. Comparing the fermentation quality among all silages in the present study, the combined additives SM and EM performed best in improving the silage quality of napiergrass.

In this study, the effects of epiphytic microbiota from different forages on the fermentation characteristics and microbial community structure of forage sorghum silage were investigated. The gamma irradiated sterilized forage sorghum was treated through sterile water, epiphytic microbiota of forage sorghum (FSm), Sudan grass (SDm), Napier grass (NPm), and maize (MZm). NPm and SDm inoculated silages showed similar pH value and lactic acid (LA) and acetic acid (AA) contents at day 3 and 60 of ensiling. The final silage of FSm and MZm showed lower (p < 0.05) pH and AA content and a higher LA content compared to the NPm and SDm silages. Bacterial species from the Weisella genus were predominantly present in FSm, NPm, and SDm, while Lactococcus dominated the MZm silage during early ensiling. Lactobacillus was predominant in all inoculated terminal silages. Overall, the four inoculated microbiota decreased the pH value of silage and were dominated by lactic acid bacteria (LAB); however, the NPm and SDm treatments resulted in comparatively higher AA contents which could have an inhibitory effect on the secondary fermentation developed by the yeast and enhanced the aerobic stability of forage sorghum silage.

Objective To study the contribution of plant enzyme and microbial activities on protein degradation in silage, this study evaluated the nitrogen transformation dynamics during ensiling of non- and irradiated alfalfa (Medicago sativa L.) and red clover (Trifolium pratense L.). Methods Alfalfa and red clover silages were prepared and equally divided into two groups. One group was exposed to γ-irradiation at a recommended dosage (25 Gky). Therefore, four types of silages were produced: i) non-irradiated alfalfa silage; ii) irradiated alfalfa silage; iii) non-irradiated red clover silage; and iv) irradiated red clover silage. These silages were opened for fermentation quality and nitrogen components analyses after 1, 4, 8, and 30 days, respectively. Results The γ-irradiation successfully suppressed microbial activity, indicated by high pH and no apparent increases in fermentation end products in irradiated silages. All nitrogen components, except for peptide-N, increased throughout the ensiling process. Proteolysis less occurred in red clover silages compared with alfalfa silages, indicated by smaller (p

Objective This study was to evaluate the fermentation dynamics, structural and nonstructural carbohydrate composition and in vitro gas production of rice straw ensiled with lactic acid bacteria and molasses. Methods Fresh rice straw was ensiled in 1-L laboratory silos with no additive control (C), Lactobacillus plantarum (L), molasses (M) and molasses+Lactobacillus plantarum (ML) for 6, 15, 30, and 60 days. After storage, the silages were subjected to microbial and chemical analyses as well as the further in vitro fermentation trial. Results All additives increased lactic acid concentration, and reduced pH, dry matter (DM) loss and structural carbohydrate content relative to the control (p

To elucidate the differences of various sugar sources and doses on rice straw silage, the effect of molasses (M, direct substrate) and fibrolytic enzyme (E, indirect substrate) with four doses on fermentation dynamics, carbohydrates and in vitro digestibility was evaluated. Molasses (0, 2, 3 and 4% of fresh weight) or fibrolytic enzyme (0, 0.02, 0.04 and 0.06%) were applied to fresh rice straw ensiling for 3, 6, 9, 15, 30 and 60 days. All additives increased lactic acid content and lactic/acetic acid, reduced pH, NH3-N content of resulting silage (p

In order to enhance the feed’s nutritive value and utilisation efficiency of local straws in Tibet, hulless-barley straw was conserved as silage by mixing with tall fescue and adding the tested additives and their combination. In experiment 1, hulless-barley straw was ensiled with four levels of tall fescue (0, 20, 40 or 60 of fresh weight, FW) in laboratory silos (1 L) for 60 days. The results suggested that 40% hulless-barley straw inclusion of 60% tall fescue was proper for further study. In experiment 2, a mixture of hulless-barley straw and tall fescue (40/60) were treated with (1) no additive (control), (2) fibrolytic enzymes (E, 0.1% FW), (3) molasses (M, 4% FW), (4) Lactobacillus plantarum MTD-1 (Lp, 106 cfu g−1 FW), (5) E + Lp and (6) M + Lp, respectively. Silos were opened after 7, 24, 45 and 60 days of ensiling respectively. The fermentation quality and structural carbohydrates were analysed. After 60 days of ensiling, all the additives except fibrolytic enzymes increased the lactic acid contents, and decreased the pH, ammonia nitrogen and butyric acid contents. All the treated silages had lower neutral detergent fibre, acid detergent fibre, hemicellulose and cellulose contents than that of control, especially in E + Lp and M + Lp treated silages. In conclusion, application of L. plantarum together with fibrolytic enzymes or molasses could further improve the fermentation quality and structural carbohydrate compositions in comparison with using each of the three additives alone.Highlights Enhancing the utilisation efficiency of local straws by mixed silage. Improving the feed’s nutritive value by reducing the crude fibre contents.

Fatty liver disease is one of the most common disorders afflicting dairy cows during the postpartum period, and is associated with increased blood non-esterified fatty acid (NEFA) uptake by the liver. Major long-chain fatty acids (LCFA) in NEFA are palmitic (PA), palmitoleic (POA), stearic (SA), oleic (OA), and linoleic (LA) acid. In order to investigate the characteristics of lipid accumulation and injury caused by these NEFA, primary calf hepatocytes were isolated and challenged for 12 h with 1.2 mmol/L PA, POA, SA, OA, LA, or a mixture of these LCFA (NEFA). Compared with POA, OA, and LA, culture with PA and SA led to greater abundance of CCAAT-enhancer binding protein, glucose-regulated protein 78 mRNA, and stearoyl-CoA desaturase 1 mRNA along with greater concentrations of H2O2, malondialdehyde and reactive oxygen species (ROS). Although culture with POA, OA, and LA led to lower very low density lipoprotein (VLDL) concentration in cell culture medium, POA and OA led to greater concentrations of triacylglycerol, protein abundance of sterol regulatory element-binding protein 1c, fatty acid synthase, acetyl coenzyme A carboxylase 1, ApoB100, and sortilin 1 (SORT1). Compared with individual fatty acids, culture with NEFA led to an intermediate degree of lipid accumulation and hepatocytes damage. Overall, the data suggest that saturated fatty acids cause more severe oxidative and ER stress. However, unsaturated fatty acids cause serious lipid accumulation. Furthermore, a fatty acid balanced nutrient regulation was suggested useful improve liver health of transition period dairy cows.

The aim of this study was to determine the effect of applying three of lactic acid bacteria isolates AZZ1, AZZ4, AZZ7 and one commercial (CB) on the fermentation dynamics and aerobic stability of Napier grass silage. The grass was ensiled with or without the addition of a lactic acid bacteria inoculant at 1.0×106 CFU/g of fresh material to Napier grass silage. Napier grass was ensiled in laboratory silos and treated as follows: no additives (control), Pediococcus acidilactici (AZZ1), Lactobacillus plantarum subsp. plantarum (AZZ4), Lactobacillus plantarum subsp. argentoratensis (AZZ7) and Lactobacillus plantarum, Ecosyl MTD/1 (CB). To follow the fermentation dynamics during ensiling samples were taken on days 7, 14, 30, 60 and 90 of ensiling for chemical and microbiological analysis. Aerobic stability was determined on day 90 of ensiling. The experimental design was a completely randomized design, with a 5 × 5 × 3. The inoculant resulted in a more decreased (P

Fibroblast Growth Factor 21 (FGF21) is a novel metabolic regulator involved in lipid utilization, and it can involve in the regulation of lipid metabolism. To determine the physiological function of FGF21 in relation to a high-fat diet (HFD) and high cholesterol, the effect of FGF21 on immune response indicators and hematological parameters was investigated. In the experiment, a total of 24 female mice were selected, 12 of which were fed with HFDs (group 1) and the other were fed with normal diet (group 2). After 30 days, the mice of two groups were respectively divided into two groups on average: 1. HFDs mice were used as model group; 2 normal diet mice were used as control group; 3. HFDs mice injection were used as model + FGF21 group; 4. normal diet injection were used as control+FGF21 group. Mice were sacrificed at 3-day intervals and the livers were isolated and analyzed. Blood was collected and analyzed for CD3 and CD19 by flow cytometry and IL-4, IL-6 and TNF-α by ELISA. Serum TG, TC, NEFA, LDL-c, and HDL-c levels were measured by automatic biochemical analyzer. Although CD3 and CD19 varied, the difference was not significant, and the levels of IL-4, IL-6, TNF-α, TC, TG LDL-c, and NEFA in the model exogenous injection FGF21 group were lower than in the model group (P

The purpose of this study is to isolate and identify lactic acid bacteria from elephant grass (Pennisetum purpureum) silage and examine their effect on the silage fermentation quality. Three of lactic acid bacteria strains were isolated from elephant silage and their characterization, identification, and influence on silage quality after 90 d of ensiling was studied. All three strains were Gram-positive, catalase-negative, and were grown in 6.5% NaCl and pH 4.00. Strains AZZ1, was identified as for genus pediococcus, whereas, AZZ4 and AZZ7 strains were classified as genus Lactobacillus according to the phenotype, 16S rRNA, and RecA gene analysis. Three strains were used as additives at 1.0×106 CFU/g of fresh material of elephant grass. Strain AZZ4 is found to be the most effective in improving the fermentation quality of the elephant grass silage, as indicated by a lower (P

Objective To assess the potency of calcium propionate (CAP) used as silage additive, an experiment was carried out to evaluate the effect of CAP on the nitrogen transformation, fermentation quality and aerobic stability of alfalfa silages. Methods Alfalfa was ensiled with four levels of CAP (5, 10, 15, and 20 g/kg of fresh weight [FW]) in laboratory silos for 30 days. After opening, the silages were analyzed for the chemical and microbiological characteristics, and subjected to an aerobic stability test. Results The increasing proportion of CAP did not affect pH, lactic acid (LA) concentrations and yeast counts, while linearly decreased counts of enterobacteria (p = 0.029), molds (p

Objective Four lactic acid bacteria (LAB) strains isolated from common vetch, tall fescue and perennial ryegrass on the Tibetan Plateau were characterized, and their effects on the fermentation quality of Italian ryegrass (Lolium multiflorum Lam.) silage were studied. Methods The four isolated strains and one commercial inoculant (G, Lactobacillus plantarum MTD-1) were evaluated using the acid production ability test, morphological observation, Gram staining, physiological, biochemical and acid tolerance tests. The five LAB strains were added to Italian ryegrass for ensiling at three different temperatures (10°C, 15°C, and 25°C). Results All isolated strains (LCG3, LTG7, I5, and LI3) could grow at 5°C to 20°C, pH 3.0 to 8.0 and NaCl (3.0%, 6.5%). Strains LCG3, LTG7, I5, and LI3 were identified as Lactobacillus plantarum, Pediococcus acidilactici, Lactobacillus paraplantarum, and Lactobacillus casei by sequencing 16S rDNA, respectively. All LAB inoculants significantly (p

Freezing damages in forages represents a major economic loss to agriculture. This study was conducted to investigate the effects of freeze–thaw (FT) event on microbial community dynamics of red clover silage. Results showed that the FT-treated material displayed higher proportions of Weissella and aerobic bacteria, while lower Pantoea and Enterobacter compared with the control material. The FT event promoted the development of Lactobacillus in silage microflora, inducing more intense lactic fermentation after an initial short lag. The aerobic bacteria were suppressed immediately after the onset of ensiling. Microbiomes of the two silages tended to be almost similar after 2 days of ensiling. However, a small number of aerobic bacteria tended to revitalize in the FT silage with prolonged ensiling time, indicated by apparent abundances of Acinetobacter and Pseudomonas at the end of ensiling. The results obtained here suggest that the FT event could promote the development of Lactobacillus during ensiling and the control of aerobe revitalization need to be concerned with silages made from the freeze-damaged forages.

ABSTRACT: The objective of present study was to investigate the dynamics of change in fermentation and fatty acid profiles in high moisture (DM=232g/kg FW) alfalfa silages during ensiling at 45°C, 30°C and 15°C. After ensiling for 1, 3, 7, 21, 39 and 65 days, silages was sampled and examined. Lactic fermentation changing into butyric fermentation in silage ensiled at 30°C and 45°C occurred on ensiling for 21 and 65 days, respectively, as accompanied with high ammonia-N content ( >150g/kg N), which subsequently resulted in a sudden increase of pH (P < 0.05). In contrast, the increase of lactic acid content was observed in silage ensiled at 15°C during ensiling for 65 days (P

Aflatoxin B1 (AFB1), a mycotoxin found in food and feed, is immunotoxic to animals and poses significant threat to the food industry and animal production. The primary target of AFB1 is the liver. To overcome aflatoxin toxicity, probiotic-mediated detoxification has been proposed. In the present study, to investigate the protective effects and molecular mechanisms of Lactobacillus bulgaricus or Lactobacillus rhamnosus against liver inflammatory responses to AFB1, mice were administered with AFB1 (300 μg/kg) and/or Lactobacillus intragastrically for 8 weeks. AML12 cells were cultured and treated with AFB1, BAY 11-7082 (an NF-κB inhibitor), and different concentrations of L. bulgaricus or L. rhamnosus. The body weight, liver index, histopathological changes, biochemical indices, cytokines, cytotoxicity, and activation of the NF-κB signaling pathway were measured. AFB1 exposure caused changes in liver histopathology and biochemical functions, altered inflammatory response, and activated the NF-κB pathway. Supplementation of L. bulgaricus or L. rhamnosus significantly prevented AFB1-induced liver injury and alleviated histopathological changes and inflammatory response by decreasing NF-κB p65 expression. The results of in vitro experiments revealed that L. rhamnosus evidently protected against AFB1-induced inflammatory response and decreased NF-κB p65 expression when compared with L. bulgaricus. These findings indicated that AFB1 exposure can cause inflammatory response by inducing hepatic injury, and supplementation of L. bulgaricus or L. rhamnosus can produce significant protective effect against AFB1-induced liver damage and inflammatory response by regulating the activation of the NF-κB signaling pathway.