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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

The objective of this study was to investigate the effect of lactic acid bacteria (LAB) inoculums on fermentation quality and in vitro digestibility of corn stover silage. Corn stover was ensiled without (control) or with Lactobacillus plantarum (LP), Enterococcus faecalis (EF), and Enterococcus mundtii (EM) for 45 days. The fermentation characteristics were assessed, and subsequent in vitro dry matter digestibility (DM-D), neutral detergent fiber digestibility (NDF-D), volatile fatty acids (VFA), methane (CH4) production, cellulolytic bacteria proportions and their activities per corn stover silage were also determined. There was no significant difference (P>0.05) among the silage pH, lactic acid, crude protein (CP), water soluble carbohydrates (WSC) and lignocelluloses contents of different treatments. The relative proportions of Ruminococcus flavefaciens and Fibrobacter succinogenes, carboxymethyl-ocellulose and β-glycosidase activities, DM-D, NDF-D, and VFA production of in vitro incubation was higher (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.

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.

The prognosis for endocrine-independent prostate carcinoma is still poor due to its highly metastatic feature. In the present work, TPX2 (the targeting protein for Xklp2), which is known as a micro-tubulin interacted protein, was identified as a novel coactivator of ETS-1, a transcription factor that plays a central role in mediating the metastasis of human malignancies. TPX2 enhanced the transcription factor activation of ETS-1 and increased the expression of ETS-1’s downstream metastasis-related genes, such as mmp3 or mmp9, induced by HGF (hepatocyte growth factor), a typical agonist of the HGF/c-MET/ETS-1 pathway. The protein-interaction between TPX2 and ETS-1 was examined using immunoprecipitation (IP). TPX2 enhanced the accumulation of ETS-1 in the nuclear and the recruitment of its binding element (EST binding site, EBS) located in the promoter region of its downstream gene, mmp9. Moreover, TPX2 enhanced the in vitro or in vivo invasion of a typical endocrine-independent prostate carcinoma cell line, PC-3. Therefore, TPX2 enhanced the activation of the HGF/ETS-1 pathway to enhance the invasion of endocrine-independent prostate carcinoma cells and thus it would be a promising target for prostate carcinoma treatment.

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

Abstract Background Butyric fermentation and a substantial loss of dry matter (DM) often occur in alfalfa silage during the rainy season, which is not conducive to subsequent biofuel production. Currently, there have been negative effects on the combination of cellulases and lactic acid bacteria (LAB) on processing high-moisture alfalfa silage; however, transgenically engineered LAB strains that secrete cellulase have been proposed as an alternative approach to avoid the above problem. The objective of the present study was to construct engineered Lactococcus lactis strains with high-efficiency secretory-expressing cellulase genes from Trichoderma reesei and to investigate the effects of the combination of transgenically engineered L. lactis strains HT1/pMG36e-usp45-bgl1, HT1/pMG36e-usp45-cbh2, and HT1/pMG36e-usp45-egl3 (HT2) on fermentation quality, structural carbohydrate degradability and nonstructural carbohydrate fermentation kinetics of high-moisture alfalfa silage treated without additive as a negative control (Control), or/and with cellulase (EN), wild-type L. lactis subsp. lactis MG1363 (HT1) and the combination of HT1 and EN (HT1 + EN) as positive additive controls. Results Engineered L. lactis strains were successfully constructed and efficiently secreted endoglucanase (1118 mU/mL), cellobiohydrolase (222 mU/mL), and β-glucosidase (131 mU/mL) and had high filter paper activity (236 mU/mL). Ensiling experiments verified that HT2 obtained the highest fermentation quality score (83.6) and most efficiently processed high-moisture alfalfa silage, demonstrated by a low pH (4.49) and ammonia-N content (106 g/kg nitrogen) and a high lactic acid content (67.1 g/kg DM) and without butyric acid. Change curves of structural carbohydrates revealed that HT2 degraded more lignocelluloses, demonstrated by the lowest contents of neutral detergent fibre, acid detergent fibre, cellulose and hemicellulose after ensiling for 60 days. Kinetic analysis showed that the most residual water-soluble carbohydrates, glucose, fructose and xylose generated by lignocellulose degradation were produced by HT2, followed by HT1 + EN. The HT2-treated silages had the highest DM recovery, had the fewest Clostridia spores, emitted a fragrance and were not sticky. Conclusion HT2 improved the conversion of lignocellulose to sugars and processed high-moisture alfalfa silage efficiently. This is a novel strategy that can be used to enhance lignocellulosic degradation in high-moisture alfalfa via a bioprocess with transgenically engineered L. lactis strains, which could enhance the development of alfalfa as a biomass feedstock and promote second-generation biofuel development in the rainy season.

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.

Prostate cancer remains a significant cause of cancer-related deaths in male population. More recently, accumulating evidence continues to implicate long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and mRNAs in various types of cancers, including prostate cancer. The current study aimed to elucidate the role of lncRNA AGAP2-AS1/miR-195-5p/PDZ and LIM domain 5 (PDLIM5) in prostate cancer progression. Initially, microarray expression profiles were applied to screen differentially expressed lncRNAs/miRNAs/genes associated with prostate cancer. Dual-luciferase reporter and RNA pull-down/RIP assays were subsequently performed to explore the interactions among lncRNA AGAP2-AS1, miR-195-5p, and PDLIM5, after which their expression was detected in cancer tissues and cells. Next, gain- and loss-of-function approaches were employed to elucidate the mechanism of lncRNA AGAP2-AS1/miR-195-5p/PDLIM5 in the processes of cell proliferation, migration and invasion as well as tumor growth. LncRNA AGAP2-AS1 was found to be highly expressed in prostate cancer. Silencing of lncRNA AGAP2-AS1 contributed to the suppression of proliferation, migration and invasion of cancer cells in vitro. Besides, lncRNA AGAP2-AS1 could bind to miR-195-5p which targeted PDLIM5 and subsequently downregulated its expression, ultimately impeding the progression of prostate cancer. Additionally, lncRNA AGAP2-AS1 inhibition led to an up-regulated expression of miR-195-5p and down-regulated PDLIM5 expression, resulting in delayed tumor growth in vivo. Taken together, the key findings of our study demonstrated that lncRNA AGAP2-AS1 silencing exerted suppressive effects on the development of prostate cancer via the miR-195-5p-dependent downregulation of PDLIM5. Our findings highlighted the potential of lncRNA AGAP2-AS1 as a promising novel molecular target for prostate cancer therapy.

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

This study was conducted to evaluate the effect of lactic acid bacteria (LAB) on fermentation quality, mycotoxin concentrations, and microbial communities of whole-crop corn silages infested with mycotoxigenic fungi. Cultured spores (106 cfu/mL) of mycotoxigenic Aspergillus flavus and Fusarium graminearum were sprayed (5 mL) on corn forage on 27 July and 10 August 2018. On 21 August 2018, sprayed (FI; 3 plots) and unsprayed (NFI; 3 plots) corn forage were harvested at the 1/2 kernel milk line stage, followed by chopping and ensiling without inoculants (CON), or with Lactobacillus buchneri (LB, 1 × 106 cfu/g FW), Lactobacillus plantarum (LT, 1 × 106 cfu/g FW), or L. buchneri + L. plantarum (BT: both L. buchneri and L. plantarum applied at 0.5 × 106 cfu/g FW). After 90 d of ensiling, FI silages had a higher (p < 0.05) pH value and higher acetic acid (ACA), ethanol, and ammonia nitrogen (ammonia N) concentrations, but lower (p < 0.05) lactic acid (LA) concentrations than NFI silage. The inoculants decreased pH and increased LA concentration and LA/ACA compared with CON. The aflatoxin B1 (AFB1) was only detected in FI fresh corn and silages; ensiling decreased (p < 0.05) AFB1 concentration compared with fresh corn, and LB and BT decreased AFB1 concentration compared with CON. The zearalenone (ZEN), deoxynivalenol (DON), and fumonisin B1 (FB1) concentrations were similar (p < 0.05) for NFI silages, while ZEN concentration in BT was the lowest (p < 0.05) among all FI silages; DON and FB1 concentrations in LB, LT, and BT silages were significantly lower (p < 0.05) than those of CON in FI silages. The fungal infestation increased the bacterial and fungal diversity of silages compared with NFI silages. The FI silages had a higher relative abundance (RA) of Lactobacillus, Weissella, Wickerhamomyces, Pichia, and Epicoccum than the corresponding NFI silages. The RA of Aspergillus and Fusarium markedly decreased after 90 d of ensiling, and the inoculation expanded this trend irrespective of fungal infestation. The Penicillium in FI silages survived after 90 d of ensiling, while the inoculants decreased the RA of Penicillium. Inoculants mitigate the adverse effects of fungal infestation on corn silage quality by changing the bacterial and fungal communities.

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

This study was conducted to evaluate the effect of lactic acid bacteria and propionic acid on the fermentation quality, aerobic stability and in vitro gas production kinetics and digestibility of whole-crop corn based total mixed ration (TMR) silage. Total mixed ration was ensiled with four treatments: (1) no additives (control); (2) an inoculant (Lactobacillus plantarum) (L); (3) propionic acid (P); (4) propionic acid+lactic acid bacteria (PL). All treatments were ensiled in laboratory-scale silos for 45 days, and then subjected to an aerobic stability test for 12 days. Further, four TMR silages were incubated in vitro with buffered rumen fluid to study in vitro gas production kinetics and digestibility. The results indicated that all TMR silages had good fermentation characteristics with low pH (

Objective The objective of this study was to evaluate the effect of sodium diacetate (SDA) on fermentation profile, chemical composition and aerobic stability of alfalfa (Medicago sativa L.) silage. Methods Fresh alfalfa was ensiled with various concentrations of SDA (0, 3, 5, 7, and 9 g/kg of fresh forage). After 60 days of the ensiling, the samples were collected to examine the fermentative quality, chemical composition and aerobic stability. Results The application of SDA significantly (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

Supershort avalanche electron beam (SAEB) plays an important role in nanosecond-pulse discharges. This paper aims at reviewing experiments results on characteritics of SAEB and its spectra in different gases in nanosecond-pulse discharges. All the joint experiments were carried in the Institute of High Current Electronics of the Russian Academy of Sciences and the Institute of Electrical Engineering of the Chinese Academy of Sciences. In these experiments, the generation of a SAEB in SF6 in an inhomogeneous electric field was studied on three generators with pulse rise times of 0.3, 0.5 and ∼2 ns. Firstly, the comparison of SAEB parameters in SF6 with those obtained in other gases (air, nitrogen, argon, and krypton) is introduced. Secondly, the SAEB spectra in SF6 and air at pressures of 10 kPa (75 torr), and 0.1 MPa (750 torr) are reviewed and discussed. Finally, 1.5-D theoretical simulation of the supershort pulse of the fast electron beam in a coaxial diode filled with SF6 at atmospheric pressure is described. The simulation was carried out in the framework of hybrid model for discharge and runaway electron kinetics. The above research progress can provide better understanding of the investigation into the mechanism of nanosecond-pulse discharges.

This experiment was conducted to study the effect of molasses on the fermentation characteristics of mixed silage ensiled rice straw and vegetable by-products with alfalfa. Mixture (202 g kg−1 dry matter (DM)) consisting of rice straw, broccoli residue and alfalfa at the ratio of 5:4:1 was ensiled with three experimental treatments: (1) no additives (control); (2) molasses at 2.5% (M1); (3) molasses at 5% (M2) on a fresh matter basis of mixture, respectively. All treatments were packed into laboratory-scale silos, and three silos per treatment were sampled on days 1, 3, 5, 14 and 30. The result showed that the pH value of all mixed silages decreased gradually with the time of ensiling except for the control silage, in which a significant increase (P

Background: Cold-active endo-1, 4-β-glucanase (EglC) can decrease energy costs and prevent product denaturation in biotechnological processes. However, the nature EglC from C. farmeri A1 showed very low activity (800 U/L). In an attempt to increase its expression level, C. farmeri EglC was expressed in Escherichia coli as an N-terminal fusion to protein S (ProS) from Myxococcus xanthus. Results: A novel expression vector, pET(ProS-EglC), was successfully constructed for the expression of C. farmeri EglC in E. coli. SDS-PAGE showed that the recombinant protein (ProS-EglC) was approximately 60 kDa. The activity of ProS-EglC was 12,400 U/L, which was considerably higher than that of the nature EglC (800 U/L). ProS-EglC was active at pH 6.5–pH 8.0, with optimum activity at pH 7.0. The recombinant protein was stable at pH 3.5–pH 6.5 for 30 min. The optimal temperature for activity of ProS-EglC was 30°C–40°C. It showed greater than 50% of maximum activity even at 5°C, indicating that the ProS-EglC is a cold-active enzyme. Its activity was increased by Co2+ and Fe2+, but decreased by Cd2+, Zn2+, Li+, methanol, Triton-X-100, acetonitrile, Tween 80, and SDS. Conclusions: The ProS-EglC is promising in application of various biotechnological processes because of its cold-active characterizations. This study also suggests a useful strategy for the expression of foreign proteins in E. coli using a ProS tag.

Space charge injected into a liquid dielectric by an electrode under an impulse voltage causes the electric field to be distorted, which affects the dielectric's insulation performance. In this study, the authors study the surface modification of three-electrode materials, namely aluminium, copper, and stainless steel, by sputtering of titanium dioxide (TiO(2)) based on magnetron sputtering. The breakdown voltage of propylene carbonate and the surface morphology of the electrodes before and after modification were examined. Furthermore, the distribution of the space charge injected into the propylene carbonate from the different electrode materials was also measured based on the Kerr electro-optic effect. These results indicate that the breakdown voltage of propylene carbonate increased by 7, 4, and 9% after surface modification of aluminium, copper, and stainless steel, respectively. After sputtering of TiO(2), micro-grooves on the surface of the three-electrode materials became smooth, and the surface roughness decreased, which changed the barrier height of the solid–liquid interface. The space charge injection mode of the three-electrode materials before and after surface modification was bipolar injection; however, the space charge injection amounts were markedly lower for the modified electrodes.

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 aim of the current study was to investigate the effects of a lactic acid bacteria inoculant (Lactobacillus plantarum, LP), fibrolytic enzyme (EN), combination of LP and EN (LP+EN) on fermentation quality, nutritive characteristics and in vitro digestibility of total mixed ration (TMR) silages containing 0, 7.5 and 15.0% (on dry matter basis) of rape (Brassica campestris L.) straw (RS) (denoted as CTMR, LTMR and MTMR, respectively). After ensiling for 60 days, TMR silages without additives were well preserved, but MTMR had higher pH than CTMR and LTMR. There were no differences in other parameters of fermentation quality, microbial composition, nutrition and in vitro digestibility between CTMR and LTMR except for yeast and mold number and crude protein (CP) content. CTMR and LTMR silage had higher CP content, in vitro neutral detergent fiber digestibility (IVNDFD) and in vitro acid detergent fiber digestibility (IVADFD), lower acid detergent fiber (ADF) content than MTMR silage. LP and EN decreased pH and increased dry matter (DM) recovery of TMR silages. LP+EN improved the fermentation quality, nutritive characteristics and in vitro digestibility of TMR silages, showed by lower pH, ADF content, higher lactic acid content, in vitro dry matter digestibility (IVDMD) and IVNDFD. Therefore, It was suggested that TMR silage contained 7.5% RS on a DM basis and treated with LP+EN can be as a useful feed for ruminant.

Understanding the mechanism of dielectric breakdown is important for improving the breakdown field of a polymer. In this work, dielectric barrier discharge (DBD) treatment was applied to one surface of P(VDF-CTFE) (vinylidene fluoride-chlorotrifluoroethylene) film, and the dielectric properties of the film were studied. When the treated surface was connected to the high potential side of the power source for the breakdown test, the breakdown field of the treated film was significantly reduced compared to that of the pristine film. Based on the characterization results for the surface chemistry and morphology, it was proposed that the phenomenon was caused by the combined effects of hole injection from the metal electrode and the damage of polymer chains near the surface of the polymer film after the DBD treatment process.

Pulsed discharge plasma and its application is one of the promising directions in civilian areas of pulsed power technology. In order to promote the research and development of the theory and application technology for pulsed discharge plasma, in this paper, recent progress on the mechanism of nanosecond-pulse gas discharge and the characteristics and applications of typical pulsed plasma at the Institute of Electrical Engineering, Chinese Academy of Sciences is reviewed. Firstly, progress on mechanism of nanosecond-pulse discharge based on runaway electrons and measurement technology of runaway electrons is introduced. Then, the characteristics of three typical discharges, including direct-driven pulsed discharge, pulsed dielectric barrier discharge and pulsed plasma jet, are reviewed. Furthermore, typical plasma applications of pulsed plasma on surface modification and methane conversion are presented.

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

This study deals with experimental and theoretical simulation data showing the influence of electrode spacing and gas pressure on parameters of a supershort avalanche electron beam (SAEB) formed in SF(6) and nitrogen at different rise times and amplitudes of a voltage pulse. Using GIN-55-01, VPG-30-200, and SLEP-150M pulsers, tubular cathodes with a diameter of 6 mm, as well as gaps of 3, 5, and 8 mm, it was shown that the SAEB current amplitude can both increase and decrease depending on an electrode spacing, a waveform and a rise time of the voltage pulse, as well as the pressure of SF(6) and nitrogen. It was established as a result of simulation that maximal voltage across the gap during the process of generation of runaway electrons and the thickness of an anode foil have a major effect on the SAEB current pulse amplitude.

The objective of this study was to evaluate the effects of molasses and propionic acid on the fermentation quality and aerobic stability of total mixed ration (TMR) silages prepared with whole-plant corn in Tibet. TMR (354 g/kg DM) was ensiled with four different treatments: no additive (control), molasses (M), propionic acid (P), and molasses+propionic acid (PM), in laboratory silos (250 mL) and fermented for 45 d. Silos were opened and silages were subjected to an aerobic stability test for 12 days, in which chemical and microbiological parameters of TMR silages were measured to determined the aerobic deterioration. After 45 d of ensiling, the four TMR silages were of good quality with low pH value and ammonia/total N (AN), and high lactic acid (LA) content and V-scores. M silage showed the highest (p105 cfu/g FM), however, it appeared to be more stable as indicated by a delayed pH value increase. P and PM silages showed fewer yeasts (

Background Many biotechnological and industrial applications can benefit from cold-adapted EglCs through increased efficiency of catalytic processes at low temperature. In our previous study, Citrobacter farmeri A1 which was isolated from a wood-inhabiting termite Reticulitermes labralis could secrete a cold-adapted EglC. However, its EglC was difficult to purify for enzymatic properties detection because of its low activity (0.8 U/ml). The objective of the present study was to clone and express the C. farmeri EglC gene in Escherichia coli to improve production level and determine the enzymatic properties of the recombinant enzyme. Methods The EglC gene was cloned from C. farmeri A1 by thermal asymmetric interlaced PCR. EglC was transformed into vector pET22b and functionally expressed in E. coli. The recombination protein EglC22b was purified for properties detection. Results SDS-PAGE revealed that the molecular mass of the recombinant endoglucanase was approximately 42 kDa. The activity of the E. coli pET22b-EglC crude extract was 9.5 U/ml. Additionally, it was active at pH 6.5–8.0 with an optimum pH of 7.0. The recombinant enzyme had an optimal temperature of 30–40 °C and exhibited >50% relative activity even at 5 °C, whereas it lost approximately 90% of its activity after incubation at 60 °C for 30 min. Its activity was enhanced by Co2+ and Fe3+, but inhibited by Cd2+, Zn2+, Li+, Triton X-100, DMSO, acetonitrile, Tween 80, SDS, and EDTA. Conclusion These biochemical properties indicate that the recombinant enzyme is a cold-adapted endoglucanase that can be used for various industrial applications.

This study aims to evaluate the fermentation characteristics of Lactobacillus plantarum and Pediococcus spp isolated from sweet sorghum silage to enhance the fermentation quality of Napier grass and sweet sorghum silage. Based on molecular 16S ribosomal ribonucleic identification the isolated strains were phylogenetically related to Lactobacillus plantarum (HY1), Pediococcus acidilactici (HY2) and Pediococcus claussenii (HY3). Strains HY1, HY2 and HY3 and commercial bacteria Lactobacillus plantarum, Ecosyl; (MTD\1( were ensiled with sweet sorghum and Napier grass and the non-inoculated grasses, have been arranged in a completely randomized experimental design in a 5 (inoculants) × 3 (ensiling periods). In both grasses, the fermentation characteristics chemical composition and microbial population were assessed at 5–30 and 90 days of ensiling. The results showed that the effect of addition inoculants significantly reduced (p < 0.05) the pH, ammonia-N, acetic acid and undesirable microbial population and increased (p < 0.05) lactic acid and lactic acid bacteria counting when compared to the control. The effect of ensiling days on silage quality through the increasing lactic acid, acetic acid, ammonia-N, propionic acid and butyric acid whereas decreasing pH and water-soluble carbohydrates and microbial counts. In both sweet sorghum and Napier silage treated with isolated strains showed the best results in silage quality. The HY3 belongs to Pediococcus claussenii was not extensively studied in silage but it has shown good fermentation quality which strongly recommended to apply as probiotic.

In order to develop methods that would enlarge the feed resources in Tibet, mixtures of hulless-barley straw and tall fescue were ensiled with four levels (0, 10%, 20%, and 30% of fresh weight) of wet hulless-barley distillers’ grains (WHDG). The silos were opened after 7, 14 or 30 d of ensiling, and the fermentation characteristics and nutritive quality of the silages were analyzed. WHDG addition significantly improved fermentation quality, as indicated by the faster decline of pH, rapid accumulation of lactic acid (LA) (p

Two experiments were carried out to investigate the effects of supplemental Chinese herbs, Fructus Ligustri Lucidi (FLL), Radix Astragali (RA) and Radix Codonopsis (RC) on growth performance, blood antioxidant and immune function in Holstein dairy heifers fed high fibre diet. Experiment 1 indicated that the supplementation of the three herbs had no effect on dry matter intake. FLL Supplementation increased heifers average daily gain (ADG), final body weight and feed efficiency. Experiment 2 indicated that FLL supplementation improved the blood antioxidant function with higher concentration of superoxide dismutase (SOD) and lower concentration of malondialdehyde (MDA), and improved immune function with lower concentrations of prostaglandin E2 (PGE2) and immunoreactive fibronectin (IFN-γ). Addition of FLL increased apparent digestibility of diet’s dry matter and organic matter than the other groups. It was demonstrated that FLL supplementation improved nutrient digestion, feed efficiency, blood antioxidant function, immune and growth performance for Holstein dairy heifers.

Runaway electrons play an important role in the avalanche formation in nanosecond- and subnanosecond- pulse discharges. In this paper, characteristics of a supershort avalanche electron beam (SAEB) generated at the subnanosecond and nanosecond breakdown in sulfur hexafluoride (SF_{6}) in an inhomogeneous electric field were studied. One pulser operated at negative polarity with voltage pulse amplitude of ∼130 kV and rise time of 0.3 ns. The other pulser operated at negative polarity with voltage pulse amplitude of 70 kV and rise time of ∼1.6 ns. SAEB parameters in SF_{6} are compared with those obtained in krypton (Kr), nitrogen (N_{2}), air, and mixtures of SF_{6} with krypton or nitrogen. Experimental results showed that SAEB currents appeared during the rise-time of the voltage pulse for both pulsers. Moreover, amplitudes of the SAEB current in SF_{6} and Kr approximately ranged from several to tens of milliamps at atmospheric pressure, which were smaller than those in N_{2} and air (ranging from hundreds of milliamps to several amperes). Furthermore, the concentration of SF_{6} additive could significantly reduce the SAEB current in N_{2}-SF_{6} mixture, but it slightly affected the SAEB current in Kr-SF_{6} mixture because of the atomic/molecular ionization cross section of the gas had a much greater impact on the SAEB current rather than the electronegativity.

Extraction of polysaccharides from Gynura medica (GMPs) was optimized by response surface methodology (RSM). A central composition design including three parameters, namely extraction temperature (X1), ratio of water to raw material (X2) and extraction time (X3), was used. The best conditions were extraction temperature of 91.7 °C, extraction time of 4.06 h and ratio of water to raw material of 29.1 mL/g. Under the optimized conditions, the yield of GMPs was 5.56%, which was similar to the predicted polysaccharides yield of 5.66%. A fraction named GMP-1 was obtained after isolation and purification by DEAE-52 and Sephadex G-100 gel chromatography, respectively. GMP-1, with a molecular weight of 401 kDa, mainly consisted of galacturonic acid (GalA), xylose (Xyl), glucose (Glu). Infrared spectroscopy was used to characterize the major functional groups of GMP-1 and the results indicated that it was an acidic polysaccharide. The antioxidant and α-glucosidase inhibitory activities of GMPs and GMP-1 were determined in vitro. The results indicated that GMPs and GMP-1 show potential for use in functional foods or medicines.

ABSTRACT: Ensilage is a simple and low-cost strategy to enable long term preservation and environmentally friendly utilization of agricultural by-products, such as straws and distiller's grains (DG) for ruminants. Effect of mixing different proportions of DG and rice straw (i.e. 0, 10, 20 or 30% of DG) with or without 5% molasses addition on fermentation and chemical variables of silages was evaluated. The study was conducted as a randomized blocks design in a 4 × 2 factorial arrangement, with three replications, using laboratory silos of 1L capacity (n=24). Despite a significant interaction (P

ABSTRACT This study was designed to find the effect of vitamin E supplementation on growth, serum antioxidant enzymes, and some trace elements in Japanese quail (Coturnix coturnix japonica) under low ambient temperature. A total of 180 day-old Japanese quails were randomly divided into four groups and provided with 0 (group A), 50 (group B), 100 (group C), and 150 IU/kg (group D) vitamin E (dl-α-tocopherol acetate) under an average temperature of 9±0.5 °C for an experimental period of 42 days. The result showed that feed intake per day, body weight, weight gain per day, and feed conversion ratio did not differ significantly between the groups. Serum concentrations of superoxide and glutathione peroxidase were significantly high in birds supplemented with 150 mg/kg of vitamin E. The concentration of aspartate aminotransferase was not significantly affected between the control and treated groups; however, alanine transaminase concentration significantly reduced in group D. Zinc concentration in the blood increased significantly in group D, with no significant effect on copper and manganese between the control and treated groups. Vitamin E at the level of 150 IU/kg of feed improves the blood antioxidant status and zinc concentration, with no effect on the performance traits of quail reared under low ambient temperature.

This study proposes a novel model that integrates the generative adversarial network (GAN) with the value at risk (VaR) measuring method. The objective is to investigate the efficacy of the VaR method in addressing the issue of excessive financialization in enterprises. Firstly, the related concepts and calculation principles of VaR in the financial field are explained, and the autoregressive conditional heteroscedastic (ARCH) family-based VaR calculation method and the basic structure of GAN under the deep learning are introduced. Then, the GAN algorithm is employed to optimize and train the initialization network, transformation network, and structure network of the GAN algorithm. Finally, the optimized GAN is applied to the VaR measurement of 300 stocks in Shanghai and Shenzhen stock market. GAN demonstrates the ability to handle unbalanced data samples, sample minority data, and fit the overall distribution of minority samples. GAN introduces a groundbreaking method for data processing, and its integration with manual efforts yields significant improvements in practical applications. Moreover, (JAN demonstrates a positive impact on data set training, offering reliable potential for advancement and serving as a valuable point of reference. In conclusion, the combination of GAN under deep learning with VaR showcases a dependable practicability in assessing the risks associated with excessive financialization. [ABSTRACT FROM AUTHOR]

Objective(s): Wnt5a, which regulates the activities of osteoblasts and osteoclasts, is reportedly overexpressed in osteoarthritis (OA) tissues. The purpose of this study was to elucidate its role in the development of OA by deleting Wnt5a in osteocalcin (OCN)-expressing cells. Materials and Methods: Knee OA was induced by anterior cruciate ligament transection (ACLT) in OCN-Cre;Wnt5afl/fl knockout (Wnt5a-cKO) mice and control littermates. Eight weeks after surgery, histological changes, cell apoptosis, and matrix metabolism of cartilage were evaluated by toluidine blue, TUNEL staining, and im-immunohistochemistry analyses, respectively. In addition, the subchondral bone microarchitecture of mice was examined by micro-computed tomography (micro-CT). Results: Histological scores show substantial cartilage degeneration occurred in ACLT knees, coupled with decreased collagen type II expression and enhanced matrix metalloproteinase 13 expression, as well as higher proportions of apoptotic cells. Micro-CT results show that ACLT resulted in decreased bone mineral density, bone volume/trabecular volume, trabecular number, and structure model index of subchondral bones in both Wnt5a-cKO and control littermates; although Wnt5a-cKO mice display lower BMD and BV/TV values, no significant difference was observed between Wnt5a-cKO and control mice for any of these values. Conclusion: Our findings indicate that Wnt5a deficiency in OCN-expressing cells could not prevent an osteoarthritic phenotype in a mouse model of post-traumatic OA. [ABSTRACT FROM AUTHOR]