Responses of microbial community composition and CAZymes encoding gene enrichment in ensiled Elymus nutans to altitudinal gradients in alpine region.

High-throughput metagenomic sequence technology was employed to evaluate changes in microbial community composition and carbohydrate-active enzymes encoding gene enrichment status in Elymus nutans silages to altitudinal gradients in the world's highest alpine region of Qinghai-Tibetan Plateau (QTP). E. nutans were collected from three different altitudes in QTP: 2,600 m (low altitude), 3600 m (moderate altitude), and 4,600 m [high (H) altitude], and ensiled for 7, 14, 30, and 60 d. Results indicated an improvement in silage quality with the increasing altitude, although the acetic acid concentration and dry matter loss were greater in H altitude silages after 30 d of ensiling. Harmful bacteria or potential pathogens predominated in the microbial community on d 7 and 14 of fermentation, while genera belonging to lactic acid bacteria gradually became the main microorganisms with the increasing altitude on d 30 and 60 of ensiling. The abundance of carbohydrate-active enzymes genes responsible for macromolecular carbohydrate degradation in silage increased with increasing altitude, and those genes were mainly carried by Lactiplantibacillus and Pediococcus at 30 and 60 d of ensiling. The abundance of key enzymatic genes associated with glycolysis and organic acid production in carbohydrate metabolism pathway was higher in H altitude silages, and Lactiplantibacillus and Pediococcus were also the main hosts after 30 d of silage fermentation, except for the fact that acetic acid production was also related to genera Leuconostoc, Latilactobacillus, and Levilactobacillus. IMPORTANCE: The fermentation quality of Elymus nutans silage was getting better with the increase of altitude in the Qinghai-Tibetan Plateau. The abundance of hosts carrying carbohydrate-active enzymes genes and key enzyme genes related to organic acid production increased with increasing altitude during the later stages of fermentation. Lactiplantibacillus and Pediococcus were the core microorganisms responsible for both polysaccharide hydrolysis and silage fermentation in the late stage of ensiling. This study provided insights on the influence of different altitudes on the composition and function of silage microbiome in the Qinghai-Tibetan Plateau, and provided a reference approach for improving the quality and controllability of silage production in high altitude areas of the Qinghai-Tibetan Plateau.

A FRET based ultrasensitive fluorescent aptasensor for 6'-sialyllactose detection.

As a kind of human milk oligosaccharide, 6'-sialyllactose (6'-SL) plays an important role in promoting infant brain development and improving infant immunity. The content of 6'-SL in infant formula milk powder is thus one of the important nutritional indexes. Since the lacking of efficient and rapid detection methods for 6'-SL, it is of great significance to develop specific recognition elements and establish fast and sensitive detection methods for 6'-SL. Herein, using 6'-SL specific aptamer as the recognition element, catalytic hairpin assembly as the signal amplification technology and quantum dots as the signal label, a fluorescence biosensor based on fluorescence resonance energy transfer (FRET) was constructed for ultra-sensitive detection of 6'-SL. The detection limit of this FRET-based fluorescent biosensor is 0.3 nM, and it has some outstanding characteristics such as high signal-to-noise ratio, low time-consuming, simplicity and high efficiency in the actual sample detection. Therefore, it has broad application prospect in 6'-SL detection.

A novel dual-recognition fluorescent biosensor for sialyl-Lewisx sensitive detection.

Sialyl-Lewisx (SLex) is a tetrasugar, which plays an important role in initial inflammation and cancer cell metastasis, and can be used as a marker for cancer diagnosis and prognosis or a therapeutic target. Detecting SLex from complex biological media remains a significant challenge. Herein, a single-stranded DNA aptamer of SLex was screened based on the double-stranded DNA library-modified magnetic bead (MB)-SELEX technology. After 14 rounds of screening, 12,639 sequences were obtained and divided into nine families. Three representative sequences were selected based on the number of sequence repeats and Gibbs binding free energy, and the aptamer SLex-Apt2 with 80 nt length (Kd = 23.01 nM) had the best affinity and relatively high specificity for targeting SLex. Then, a novel dual-recognition fluorescent biosensor for SLex-sensitive detection based on aptamer SLex-Apt2 bio-dots and 3-aminobenzoboric acid-modified MB was developed. This method can detect SLex as low as 32 µM and has a good linear response in the range 100 µM to 2 mM. It has the advantages of low preparation cost, good targeting, and avoiding the occurrence of false-positive and false-negative detection results, which makes the biosensor more valuable in biological detection and clinical diagnosis.

Effects of nisin on bacterial community and fermentation profiles, in vitro rumen fermentation, microbiota, and methane emission of alfalfa silage.

BACKGROUND: Alfalfa (Medicago sativa L.) has been used widely in preparing silage. However, forage legumes are prone to contamination by spoilage bacteria during fermentation. Nisin has broad-spectrum antibacterial properties and has been applied as an inhibitor of rumen methane emissions. However, little research has been carried out on the application of nisin in silage. This study therefore aimed to investigate the impacts of different nisin concentrations on the bacterial community and fermentation dynamics, in vitro ruminal fermentation characteristics, microbiota, and methane emissions of alfalfa silage. RESULTS: The detection limits of organic acid in nisin-treated silages were not reached in 0.09 g kg-1 nisin (0.09 level) from days 1 to 7 of ensiling. With increasing nisin concentrations, the silage dry matter increased linearly (P < 0.05), and dry matter loss decreased linearly (P < 0.05). Moreover, both the 0.06 g kg-1 nisin (0.06 level) and 0.09 level treatments increased the relative abundance of Pediococcus acidilactici during ensiling. Concurrently, as the nisin concentrations increased, ruminal methane production decreased linearly (P < 0.05), while the relative abundances of ruminal Succinivibrio, Fibrobacter succinogenes and Ruminobacter amylophilus increased linearly (P < 0.05). The populations of ruminal total bacteria, methanogens, protozoa, and fungi decreased linearly with increasing nisin concentrations (P < 0.05). CONCLUSION: The addition of nisin delayed the fermentation process, preserved more nutrients in alfalfa silage, and promoted fermentation dominated by P. acidilactici in the late phase of ensiling. Moreover, nisin reduced in vitro rumen methane emissions without adverse effects on dry matter digestibility. © 2023 Society of Chemical Industry.

Effects of malic or citric acid on the fermentation quality, proteolysis and lipolysis of alfalfa silage ensiled at two dry matter contents.

This study evaluated the effects of two organic acids on the fermentation of alfalfa silages at different dry matter (DM) contents. Alfalfa was wilted to DM contents of 30% (moderately low) and 38% (normal) and ensiled without additives (control) or treated with 0.6% fresh matter DL-malic acid (MA) or 0.6% fresh matter citric acid (CA) for 60 days. After ensiling, silages with a normal DM were higher in pH, water-soluble carbohydrates (WSC) and DM loss (p < 0.05) when compared to silages ensiled at a moderately low DM. The higher DM content also limited proteolysis in silages indicated by lower concentrations of ammonia N (NH3 -N). Compared with the control group, MA and CA-treated silages had lower pH, lower concentrations of acetic acid and NH3 -N but higher concentrations of lactic acid. The addition of MA and CA reduced DM losses in silages when compared to the control group except for MA-treated silage at a moderately low DM in which only numerically lower DM loss was observed. Malic acid and CA also resulted in a higher proportion of polyunsaturated fatty acids, especially in silages with a moderately low DM. Including MA and CA could promote silage fermentation, limit proteolysis and lipolysis at the lower and medium-to-high end of DM contents in alfalfa silages.

Proteomic analysis of macrophage in response to Edwardsiella tarda-infection.

Edwardsiella tarda is an important facultative intracellular pathogen infecting a wide range of host from fish to humans. This bacterium could survive and replicate in macrophages as an escape mechanism from the host defense. E. tarda -macrophage interaction is vital in determining the outcome of edwardsiellasis. To fully elucidate the pathogenesis of E. tarda, the differential proteomes of RAW264.7 cells in response to E. tarda-infection, were analyzed at different time points with two-dimensional gel electrophoresis (2-DE) followed by liquid-chromatography-tandem mass spectrometry (LC-MS/MS) identification. 26 altered proteins (18 up-regulated and 8 down-regulated proteins) were successfully identified, which are mainly involved in formation of phagosomes, macrophage microbicidal activity and anti-apoptosis of macrophage. Moreover, 6 corresponding genes of the differentially expressed proteins were quantified by quantitative real-time PCR (qPCR) to examine the transcriptional profiles. Western blot analysis further confirmed the differential expression of 5 proteins in the proteomic profiles. Based on these findings, we hypothesize that these differentially expressed proteins likely play a pivotal role in determining the course of E. tarda-infection. The result suggested that E. tarda could develop some strategies to achieve a successful intracellular lifestyle, including modulation of phagosome biogenesis, resistance to macrophage microbicidal agent and anti-apoptosis of macrophages. Thus, this work effectively provides useful and novel protein-related information to further understand the underlying pathogenesis of E. tarda-infection.

Two nucleoside receptors from Streptomyces coelicolor: expression of the genes and characterization of the recombinant proteins.

Streptomyces coelicolor is a soil-dwelling bacterium that undergoes an intricate, saprophytic lifecycle. The bacterium takes up exogenous nucleosides for nucleic acid synthesis or use as carbon and energy sources. However, nucleosides must pass through the membrane with the help of transporters. In the present work, the SCO4884 and SCO4885 genes were cloned into pCOLADuet-1 and overexpressed in Escherichia coli BL21. Each protein was monomeric. Using isothermal titration calorimetry, we determined that SCO4884 and SCO4885 are likely nucleoside receptors with affinity for adenosine and pyrimidine nucleosides. On the basis of bioinformatics analysis and the transporter classification system, we speculate that SCO4884-SCO4888 is an ABC-like transporter responsible for the uptake of adenosine and pyrimidine nucleosides.

Analysis of Streptomyces coelicolor membrane proteome using two-dimensional native/native and native/sodium dodecyl sulfate gel electrophoresis.

Analysis of the oligomeric state of a protein may provide insights into its physiological functions. Because membrane proteins are considered to be the workhorses of energy generation and polypeptide and nutrient transportation, in this study we characterized the membrane-associated proteome of Streptomyces coelicolor by two-dimensional (2D) blue native/sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), high-resolution clear native/native PAGE, and native/SDS-PAGE. A total of 77 proteins were identified, and 20 proteins belonging to 15 complexes were characterized. Moreover, the resolution of high-resolution clear native/SDS-PAGE is much higher than that of blue native/SDS-PAGE. OBP (SCO5477) and BldKB (SCO5113) were identified as the main protein spots from the membrane fractions of S. coelicolor M145, suggesting that these two proteins are involved in extracellular peptide transportation. These two transporters exhibited multiple oligomeric states in the native PAGE system, which may suggest their multiple physiological functions in the development of S. coelicolor.

Corrigendum to "Probiotic effect of ferulic acid esterase-producing Lactobacillus plantarum inoculated alfalfa silage on digestion, antioxidant, and immunity status of lactating dairy goats" [Anim Nutr 11 (2022) 38-47].

[This corrects the article DOI: 10.1016/j.aninu.2022.06.010.].

Effects of antioxidant-rich Lactiplantibacillus plantarum inoculated alfalfa silage on rumen fermentation, antioxidant and immunity status, and mammary gland gene expression in dairy goats.

BACKGROUND: Milk synthesis in lactating animals demands high energy metabolism, which results in an increased production of reactive oxygen metabolites (ROM) causing an imbalance between oxidants and antioxidants thereby inducing oxidative stress (OS) on the animals. To mitigate OS and postpartum disorders in dairy goats and gain insight into the impact of dietary choices on redox status during lactation, a feeding trial was conducted using alfalfa silage inoculated with a high-antioxidant strain of Lactiplantibacillus plantarum. METHODS: Twenty-four Guanzhong dairy goats (38.1 ± 1.20 kg) were randomly assigned to two dietary treatments: one containing silage inoculated with L. plantarum MTD/1 (RSMTD-1), and the other containing silage inoculated with high antioxidant activity L. plantarum 24-7 (ES24-7). RESULTS: ES24-7-inoculated silage exhibited better fermentation quality and antioxidant activity compared to RSMTD-1. The ES24-7 diet elevated the total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activities in milk, serum, and feces of lactating goats (with the exception of T-AOC in milk). Additionally, the diet containing ES24-7 inoculated silage enhanced casein yield, milk free fatty acid (FFA) content, and vitamin A level in the goats' milk. Furthermore, an increase of immunoglobulin (Ig)A, IgG, IgM, interleukin (IL)-4, and IL-10 concentrations were observed, coupled with a reduction in IL-1ß, IL-2, IL-6, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α concentrations in the serum of lactating goats fed ES24-7. Higher concentrations of total volatile fatty acid (VFA), acetate, and propionate were observed in the rumen fluid of dairy goats fed ES24-7 inoculated silage. Moreover, the diet containing ES24-7 inoculated silage significantly upregulated the expression of nuclear factor erythroid 2 like 2 (NFE2L2), beta-carotene oxygenase 1 (BCO1), SOD1, SOD2, SOD3, GPX2, CAT, glutathione-disulfide reductase (GSR), and heme oxygenase 1 (HMOX1) genes in the mammary gland, while decreased the levels of NADPH oxidase 4 (NOX4), TNF, and interferon gamma (IFNG). CONCLUSIONS: These findings indicated that feeding L. plantarum 24-7 inoculated alfalfa silage not only improved rumen fermentation and milk quality in lactating dairy goats but also boosted their immunity and antioxidant status by modulating the expression of several genes related to antioxidant and inflammation in the mammary gland.

Responses of microbial community dynamics, co-occurrences, functional shifts, and natural fermentation profiles of Elymus nutans silage to altitudinal gradients.

IMPORTANCE: On the Qinghai-Tibet Plateau (QTP), feed shortages are common due to cold environmental conditions and the short growing season of crops. Therefore, effective preservation, such as the ensiling of local forage, is becoming increasingly important to balance the seasonal imbalance between the forage supply and the nutritional needs of domestic animals in this area. However, the structure of the microbial community of the forage, which is influenced by climatic conditions such as altitude differences, has a major impact on the fermentation quality and microbial succession of the ensiled forage. Therefore, we investigated microbial community dynamics, co-occurrence, functional shifts, and natural fermentation profiles of Elymus nutans silage as a function of altitudinal gradients. Results show that silage from Chenduo at higher elevations has better fermentation quality and higher abundance of Lacticaseibacillus and Levilactobacillus than ensiled forage from other regions. This work may contribute to guiding for silage production in QTP.

The Effects of Lactobacillus plantarum and Lactobacillus buchneri on the Fermentation Quality, In Vitro Digestibility, and Aerobic Stability of Silphium perfoliatum L. Silage.

In this experiment, Lactobacillus plantarum and Lactobacillus buchneri were added individually or in combination to Silphium perfoliatum L. (SP) silage to investigate the effects of different fermentation types of lactobacilli on the fermentation quality, in vitro digestibility, and aerobic stability of SP-silage, with a view to providing a certain scientific basis and technical support for obtaining high-quality SP-silage in production. The experiment comprised a non-additive group (control), an L. plantarum group (LP), an L. buchneri group (LB), and an L. plantarum and L. buchneri mixed treatment group (LPLB). Samples were taken after 60 days of fermentation and analyzed for the fermentation quality, in vitro digestibility, and aerobic stability of the SP-silage. The results showed that the addition of LP, LB, and LPLB significantly reduced the pH and proportion of ammonia nitrogen to total nitrogen and significantly increased the lactic acid, in vitro dry matter digestibility, and in vitro crude protein digestibility in the SP-silage (p < 0.05). Compared to the control group, the dry matter and crude protein contents of the LB and LPLB groups were significantly increased, while the neutral detergent fiber and acid detergent fiber contents were significantly reduced (p < 0.05). The SP-silage supplemented with LPLB had the highest dry matter and crude protein contents. The gross and digestible energies of the SP-silage in the LB and LPLB groups were significantly higher than those in the control and LP groups (p < 0.05). The aerobic stability of the SP-silage was significantly reduced by 24.14% in the LP group and increased by 58.62% and 34.48% in the LB and LPLB groups, respectively, compared to the control group (p < 0.05). It was shown that adding a combination of LP and LB resulted in the best fermentation quality, nutritional value, and in vitro digestibility of the SP-silage. LB was effective in improving the aerobic stability of SP-silage.

Effect of Inoculating Two Bacteriocin-Producing Lactiplantibacillus plantarum Strains at Ensiling on In Vitro Rumen Fermentation and Methane Emissions of Alfalfa Silage with Two Dry Matter Contents.

The objective of this study was to investigate the effects of inoculating two bacteriocin-producing strains, Lactiplantibacillus plantarum ATCC14917 and LP1-4, at ensiling on the in vitro ruminal fermentation characteristics and methane production of alfalfa silage with two dry matter (DM)contents. Before ensiling, fresh alfalfa was wilted to a moderate DM content (355 g/kg) and a high DM content (428 g/kg). The wilted alfalfa was treated with (1) distilled water (control), (2) commercial strain L. plantarum MTD/1 (MTD/1), (3) bacteriocin-producing L. plantarum ATCC14917 (ATCC14917), and (4) a bacteriocin-like substance producing L. plantarum LP1-4 (LP1-4) at 1 × 105 colony forming units (CFU)/g fresh weight. After 90 d of ensiling, the silages were used for in vitro rumen fermentation. Inoculation with the two bacteriocin-producing strains at ensiling remarkably reduced (p < 0.05) in vitro ruminal CH4 production and enhanced DM digestibility compared with the control group regardless of DM content. For silages with high DM content, inoculation with the bacteriocin-producing strains even increased (p < 0.05) in vitro ruminal total volatile fatty acid production. Therefore, the bacteriocin-producing inoculants have a great potential to mitigate ruminal methane emission but without an adverse effect on rumen fermentation of the inoculated alfalfa silage.

Current approaches on the roles of lactic acid bacteria in crop silage.

Lactic acid bacteria (LAB) play pivotal roles in the preservation and fermentation of forage crops in spontaneous or inoculated silages. Highlights of silage LAB over the past decades include the discovery of the roles of LAB in silage bacterial communities and metabolism and the exploration of functional properties. The present article reviews published literature on the effects of LAB on the succession, structure, and functions of silage microbial communities involved in fermentation. Furthermore, the utility of functional LAB in silage preparation including feruloyl esterase-producing LAB, antimicrobial LAB, lactic acid bacteria with high antioxidant potential, pesticide-degrading LAB, lactic acid bacteria producing 1,2-propanediol, and low-temperature-tolerant LAB have been described. Compared with conventional LAB, functional LAB produce different effects; specifically, they positively affect animal performance, health, and product quality, among others. In addition, the metabolic profiles of ensiled forages show that plentiful probiotic metabolites with but not limited to antimicrobial, antioxidant, aromatic, and anti-inflammatory properties are observed in silage. Collectively, the current knowledge on the roles of LAB in crop silage indicates there are great opportunities to develop silage not only as a fermented feed but also as a vehicle of delivery of probiotic substances for animal health and welfare in the future.

Lignocellulose conversion of ensiled Caragana korshinskii Kom. facilitated by Pediococcus acidilactici and cellulases.

To explore the biofuel production potential of Caragana korshinskii Kom., Pediococcus acidilactici and an exogenous fibrolytic enzyme were employed to investigate the fermentation profile, structural carbohydrates degradation, enzymatic saccharification and the dynamics of bacterial community of C. korshinskii silage. After 60 d of ensiling, all additives increased the fermentation quality. The highest lactic and acetic acids and lowest non-protein nitrogen (NPN) and ammonia nitrogen (NH3 -N) were observed in P. acidilactici and Acremonium cellulase (PA + AC) treated silage. Additionally, all additives significantly increased the ferulic acid content and fibre degradability with the highest values obtained from PA + AC silage. The bacterial community in all silages was dominated by P. acidilactici throughout the entire fermentation process. The bacterial community was also modified by the silage additives exhibiting a relatively simple network of bacterial interaction characterized by a lower bacterial diversity in P. acidilactici (PA) treated silage. The highest 6-phospho-beta-glucosidase abundance was observed in PA-treated silage at the mid-later stage of ensiling. PA treatment exhibited lower structural carbohydrates degradation but performed better in lignocellulose conversion during enzymatic saccharification. These results indicated that pretreating C. korshinskii improved its silage quality and potential use as a lignocellulosic feedstock for the production of bio-product and biofuel.

Shift of Feeding Strategies from Grazing to Different Forage Feeds Reshapes the Rumen Microbiota To Improve the Ability of Tibetan Sheep (Ovis aries) To Adapt to the Cold Season.

The dynamics of ruminant-rumen microbiome symbiosis associated with feeding strategies in the cold season were examined. Twelve pure-grazing adult Tibetan sheep (Ovis aries) (18 months old; body weight, 40 ± 0.23 kg) were transferred from natural pasture to two indoor feedlots and fed either a native-pasture diet (NPF group) or an oat hay diet (OHF group) (n = 6 per treatment), and then the flexibility of rumen microbiomes to adapt to these compositionally different feeding strategies was examined. Principal-coordinate analysis and similarity analysis indicated that the rumen bacterial composition correlated with altered feeding strategies. Microbial diversity was higher in the grazing group than in those fed with native pasture and an oat hay diet (P < 0.05). The dominant microbial phyla were Bacteroidetes and Firmicutes, and the core bacterial taxa comprised mostly (42.49% of shared operational taxonomic units [OTUs]) Ruminococcaceae (408 taxa), Lachnospiraceae (333 taxa), and Prevotellaceae (195 taxa), which were relatively stable across different treatments. Greater relative abundances of Tenericutes at the phylum level, Pseudomonadales at the order level, Mollicutes at the class level, and Pseudomonas at the genus level were observed in a grazing period than in the other two treatments (NPF and OHF) (P < 0.05). In the OHF group, due to the high nutritional quality of the forage, Tibetan sheep can produce high concentrations of short-chain fatty acids (SCFAs) and NH3-N by increasing the relative abundances of key bacteria in the rumen, such as Lentisphaerae, Negativicutes, Selenomonadales, Veillonellaceae, Ruminococcus 2, Quinella, Bacteroidales RF16 group, and Prevotella 1, to aid in nutrients degradation and energy utilization. The levels of beneficial bacteria were increased by the oat hay diet; these microbiotas are likely to help improve and maintain host health and metabolic ability in Tibetan sheep to adapt to cold environments. The rumen fermentation parameters were significantly influenced by feeding strategy in the cold season (P < 0.05). Overall, the results of this study demonstrate the strong effect of feeding strategies on the rumen microbiota of Tibetan sheep, which provided a new idea for the nutrition regulation of Tibetan sheep grazing in the cold season on the Qinghai-Tibetan Plateau. IMPORTANCE During the cold season, like other high-altitude mammals, Tibetan sheep have to adapt their physiological and nutritional strategies, as well as the structure and function of their rumen microbial community, to the seasonal variation of lower food availability and quality. This study focused on the changes and adaptability in the rumen microbiota of Tibetan sheep when they adapted from grazing to a high-efficiency feeding strategy during the cold season by analyzing the rumen microbiota of Tibetan sheep raised under the different management systems, and it shows the linkages among the rumen core and pan-bacteriomes, nutrient utilization, and rumen short-chain fatty acids. The findings from this study suggest that the feeding strategies potentially contribute to variations in the pan-rumen bacteriome, together with the core bacteriome. Fundamental knowledge on the rumen microbiomes and their roles in nutrient utilization furthers our understanding of how rumen microbial adaptation to harsh environments may function in hosts. The facts obtained from the present trial clarified the possible mechanisms of the positive effects of feeding strategy on nutrient utilization and rumen fermentation in harsh environments.

Effects of inoculating feruloyl esterase-producing Lactiplantibacillus plantarum A1 on ensiling characteristics, in vitro ruminal fermentation and microbiota of alfalfa silage.

BACKGROUND: Ferulic acid esterase (FAE)-secreting Lactiplantibacillus plantarum A1 (Lp A1) is a promising silage inoculant due to the FAE's ability to alter the plant cell wall structure during ensiling, an action that is expected to improve forage digestibility. However, little is known regarding the impacts of Lp A1 on rumen microbiota. Our research assessed the influences of Lp A1 in comparison to a widely adopted commercial inoculant Lp MTD/1 on alfalfa's ensilage, in vitro rumen incubation and microbiota. RESULTS: Samples of fresh and ensiled alfalfa treated with (either Lp A1 or Lp MTD/1) or without additives (as control; CON) and ensiled for 30, 60 and 90 d were used for fermentation quality, in vitro digestibility and batch culture study. Inoculants treated silage had lower (P < 0.001) pH, acetic acid concentration and dry matter (DM) loss, but higher (P = 0.001) lactic acid concentration than the CON during ensiling. Compared to the CON and Lp MTD/1, silage treated with Lp A1 had lower (P < 0.001) aNDF, ADF, ADL, hemicellulose, and cellulose contents and higher (P < 0.001) free ferulic acid concentration. Compared silage treated with Lp MTD/1, silage treated with Lp A1 had significantly (P < 0.01) improved ruminal gas production and digestibility, which were equivalent to those of fresh alfalfa. Real-time PCR analysis indicated that Lp A1 inoculation improved the relative abundances of rumen's total bacteria, fungi, Ruminococcus albus and Ruminococcus flavefaciens, while the relative abundance of methanogens was reduced by Lp MTD/1 compared with CON. Principal component analysis of rumen bacterial 16S rRNA gene amplicons showed a clear distinction between CON and inoculated treatments without noticeable distinction between Lp A1 and Lp MTD/1 treatments. Comparison analysis revealed differences in the relative abundance of some bacteria in different taxa between Lp A1 and Lp MTD/1 treatments. Silage treated with Lp A1 exhibited improved rumen fermentation characteristics due to the inoculant effects on the rumen microbial populations and bacterial community. CONCLUSIONS: Our findings suggest that silage inoculation of the FAE-producing Lp A1 could be effective in improving silage quality and digestibility, and modulating the rumen fermentation to improve feed utilization.

Probiotic effect of ferulic acid esterase-producing Lactobacillus plantarum inoculated alfalfa silage on digestion, antioxidant, and immunity status of lactating dairy goats.

A feeding experiment was conducted to determine the effects of inoculating alfalfa silage with a ferulic acid esterase-producing inoculum on feed digestibility, rumen fermentation, antioxidant, and immunity status of lactating dairy goats. Twenty dairy goats were distributed into 2 experimental groups consisting of control diet (Lp MTD/1, including Lactobacillus plantarum MTD/1 inoculated silage) against diet containing silage treated with ferulic acid esterase-producing L. plantarum A1 (Lp A1). Alfalfa silage inoculated with a ferulic acid esterase-producing Lp A1 had better fermentation quality than the Lp MTD/1 inoculation. The application of Lp A1 improved silage antioxidant capacity as indicated by greater total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and glutathion peroxidase (GSH-Px) activities in Lp A1 treated silage versus Lp MTD/1 treatment. Compared with Lp MTD/1 treated group, inoculation of silage with Lp A1 increased apparent total tract digestibility of dietary dry matter, organic matter and crude protein, and ruminal concentrations of total volatile fatty acids, acetate, propionate and isobutyrate as well. The results of current study also demonstrated improved antioxidant capacity and immune performance of dairy goats with Lp A1 inoculation. Feeding Lp A1-treated silage increased dairy goats' serum antioxidase activity, such as T-AOC, SOD, GSH-Px and catalase, and the serum concentration of immunoglobulin A, while decreased tumor necrosis factor α, interleukin (IL)-2 and IL-6. In addition, compared with Lp MTD/1, diet containing alfalfa silage inoculated with Lp A1 endowed dairy goats' milk with greater fat and protein contents, improved dairy goat milk quality without affecting feed efficiency.

Characterization of a novel beta-cypermethrin-degrading strain of Lactobacillus pentosus 3-27 and its effects on bioremediation and the bacterial community of contaminated alfalfa silage.

In this study, a novel beta-cypermethrin (beta-cyp)-degrading strain Lactobacillus pentosus 3-27 (LP3-27) was screened from beta-cyp-contaminated silage. The strain could degrade 96% of beta-cyp (50 mg/L) in MSM medium after 4 d of culture, while the strain lost its degradation ability when the beta-cyp concentration reached 250 mg/L. The effects of LP 3-27 on fermentation, bacterial community, and bioremediation of contaminated alfalfa silage at two dry matter (DM) contents were studied. The results showed that inoculation with LP3-27 not only degraded beta-cyp, but also improved the fermentation quality of alfalfa silage after 60 d of ensiling. Meanwhile, L. pentosus dominated the bacterial community during ensiling in LP3-27 inoculated silages, whereas Pediococcus acidilactici was the dominant species in the control silage. LP3-27 inoculation also simplified the bacterial interaction networks of ensiled alfalfa. Beta-cyp degradation was positively correlated with L. pentosus in LP- inoculated silages, which confirmed the function of beta-cyp degradation by L. pentosus. In addition, higher beta-cyp degradation was observed in silage with 35% versus 43% DM. In summary, strain LP3-27 could be used as a candidate inoculum for bioremediation of beta-cyp-contaminated silage and to produce safe silage for animal production.

The bacterial community and metabolome dynamics and their interactions modulate fermentation process of whole crop corn silage prepared with or without inoculants.

Multi-omics approach was adopted to investigate the modulation of bacterial microbiota and metabolome as well as their interactions in whole crop corn ensiling systems by inoculating homofermentative Lactobacillus plantarum or heterofermentative Lactobacillus buchneri. Inoculations of the two different inoculants resulted in substantial differences in microbial community and metabolic composition as well as their dynamics in ensiled corn. Inoculants also altered the correlations of microbiota in different manners, and various keystone species were identified in corn silages with different treatments. Many metabolites with biofunctional activities like bacteriostatic, antioxidant, central nervous system inhibitory and anti-inflammatory were found in the present silage. A constitutive difference in microbiota dynamics was found for several pathways, which were upregulated by specific taxa in middle stage of fermentation, and widespread associations between metabolites with biofunctions and the species of lactic acid bacteria dominated in silage were observed. Multiple microbial and metabolic structures and dynamics were correlated and affected the fermentation process of the corn ensiling systems. Results of the current study improve our understanding of the complicated biological process underlying silage fermentation and provide a framework to re-evaluate silages with biofunctions, which may contribute to target-based regulation methods to produce functional silage for animal production.