The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at 39°C to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at 39°C for 72 h. Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, 39°C, pH 6.5, moisture 50%, inoculum level 107 cell/g. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

Aims The aim of this research is to study the removal characteristics and evaluate the detoxify action of deoxynivalenol by Bacillus natto 16 in wheat flour as food or feed. Methods and Results The content of deoxynivalenol was determined using ELISA by testing the deoxynivalenol removal rate, and the influence of culture supernatant, intracellular substances, crude enzyme and cell wall on the deoxynivalenol in wheat flour was studied. The effect of bacterial components on the removal of deoxynivalenol was studied in the artificial gastrointestinal environment to simulate the digestion of food. Secondary metabolites were analysed by high‐performance liquid chromatography in tandem with mass spectrometry (HPLC‐MS).The cell wall can reduce the content of deoxynivalenol in the sample by adsorption, the influence of culture supernatant, intracellular substances and crude enzyme can convert deoxynivalenol into substances with a lower molecular weight. Bacterial components have no effect on deoxynivalenol in wheat flour in simulated gastric fluid and have a certain removal effect on deoxynivalenol, which is closely related to intestinal digestion time and pH, in simulated intestinal fluid. Conclusions Experimental results indicate that the removal of deoxynivalenol by Bacillus natto 16 includes adsorption and biodegradation, simulated gastric fluid would invalidate the deoxynivalenol removal activity of Bacillus natto 16’s components. Significance and Impact of Study Our study showed that as an edible probiotic bacterium, Bacillus natto 16 can effectively remove deoxynivalenol from wheat flour as food or feed, and can be used a new deoxynivalenol ‐detoxifying microbe. The results of this research could provide the theory foundation for further development and application of Bacillus natto 16.

Ruminants exhibit stronger tolerance to gossypol, an anti-nutritional factor, compared to monogastric animals. We transplanted Hu sheep rumen microbiota into male mice to investigate the role of rumen microbiota in animal gossypol tolerance. Thirty specific-pathogen-free (SPF) male C57BL/6 mice were randomly divided into three groups: normal diet (CK group), gossypol diet (FG group), and rumen microbiota transplantation (FMT group, gossypol diet). The pathological changes in the liver and small intestine of the mice, the organ coefficient, and sperm parameters were analyzed. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the blood and lactate dihydrogen-X (LDH-X) levels in the testicular tissue were also measured. The results showed that body weight, feed intake, sperm concentration, sperm motility, and LDH-X levels in the FMT group increased (p < 0.05) compared with the FG group, while the enzyme activities of ALT, AST, and AST/ALT decreased (p < 0.05). In the FMT group, the injury to liver cells was alleviated, the structure of the small intestine was intact, and the villus height and the ratio of villus height to crypt depth (V/C) were higher than those in the FG group (p < 0.05). And there were no differences in various organ coefficients and sperm deformity rates among the three groups (p > 0.05), but compared with the FG group, mice in the FMT group showed tendencies closer to those in the CK group. Rumen microbiota transplantation relieved the reproductive toxicity and liver damage induced by gossypol in male mice and improved the tolerance of recipient animals to gossypol. Additionally, rumen microbes improved the intestinal structural integrity of recipients.

Objective: This study aimed to investigate the promoting effect of a Bacillus velezensis (BV) strain on lactic acid bacteria (LAB) and determine its influence on the fermentation quality and aerobic stability of silage.Methods: Flat colony counting method was used to evaluate the effect of BV on the growth of LAB. Freshly harvested whole-plant corn was inoculated separately with BV and L. plantarum (LP), along with an uninoculated control group (CK), and assessed at 1, 3, 5, 7, 15, and 30 days of ensiling.Results: The results indicated that BV exhibited a proliferative effect on Weissella confusa, Lactobacillus plantarum L-2, and Pediococcus pentosaceus. And exhibited a more rapid pH reduction in BV-inoculated silage compared with that in CK and LP-inoculated silage during the initial stage of ensiling. Throughout ensiling, the BV and LP experimental groups showed enhanced silage fermentation quality over CK. Additionally, relative to LP-inoculated silage, BV-inoculated silage displayed reduced pH and propionic acid. BV also prolonged aerobic stability under aerobic conditions. The microbial community in BV-inoculated silage showed greater stability than that in LP-inoculated silage. Additionally, Firmicutes and Lactobacillus exhibited more rapid elevation initially in BV versus LP-inoculated silage, but reached comparable levels between the two inoculation groups in the later stage.Conclusion: In summary, BV enhanced the efficacy and aerobic stability of whole-plant corn silage fermentation by stimulating LAB proliferation.

Companion planting of white clover (Trifolium repens L.) with orchard grass (Dactylis glomerata L.), a famous hay grass, improves the forage quality of orchard grass. Microbiome profiling techniques can reveal the specific role of white clover companion planting with orchard grass. This study aimed to explore the microbiome distribution and gene functions of rhizosphere and non-rhizosphere soil via companion planting systems of white clover and orchard grass. From metagenomics sequencing analysis, we confirmed the significant role of white clover on soil environment modeling during companion planting with orchard grass. Twenty-eight biomarkers of rhizosphere soil organisms were identified during companion planting, including Proteobacteria, Betaproteobacteria, Flavobacteriia, and Caulobacterales. The number of gene functions of nitrogen and carbon fixation in companion planting was higher than that in single plants, indicating new functional flora for companion planting. We characterized specific rhizosphere effects, typical biomarker flora, and potential regulatory mechanisms for white clover-related companion planting by metagenomics analyses.

Once a silo has been opened, the silage inside will face challenges such as aerobic deterioration, rot, and contamination. Biocontrol bacteria, as a kind of biological antiseptic, are highly effective and natural and are gaining increasing attention. This study aimed to screen a strain with anti-microbial activity against silage spoilage microorganisms and examine its effects on the fermentation quality, aerobic stability, in vitro digestion, and methane production of silage. Lactic acid bacteria, pathogenic and rot-causing microorganisms, were used as indicators to screen the strains for putrefactive silage. The bacteriostatic spectrum, growth performance, and tolerance to the silage environment of the strain were tested. A strain named D-2 was screened from rotten whole-plant corn silage and identified as Bacillus velezensis through physiological and biochemical tests as well as 16S rDNA sequencing. This study found that D-2 exhibits antibacterial effects on several microorganisms, including Escherichia coli, Staphylococcus aureus, Salmonella enteritidis, Aspergillus niger, Saccharomyces cerevisiae, Fusarium oxysporum, and Fusarium graminearum. However, it has no adverse effect on Lactobacillus reuteri, Enterococcus faecium, or Lactobacillus casei. D-2 can attain a stable stage within 10 h and withstand temperatures of up to 70 °C. Moreover, this study found that D-2 had a high survival rate of over 97% after 48 h in a lactic acid environment with pH 4. Freshly chopped whole-plant corn was inoculated without or with D-2 and ensiled for 60 days. The results show that D-2 inoculations increase the content of water-soluble carbohydrates, acetic acid, and propionic acid in the silage and decrease the number of yeasts and molds, the NH₄⁺-N/TN ratio, and the pH. We also found that fermenting whole-plant corn with D-2 significantly increased the in vitro digestibility and the propionic acid content, while also significantly inhibiting methane production. After being exposed to air for 10 days, D-2 can still effectively reduce the total number of yeasts and molds, prevent the decrease in lactic acid bacteria, and inhibit the increase in the pH and NH₄⁺-N/TN ratio of silage products. Overall, D-2 is resistant to pathogenic and rot-causing microorganisms, allowing for easy adaptation to silage production conditions. D-2 can effectively improve aerobic stability and reduce losses in the nutritional value of silage, indicating possible applications for the prevention of silage rot and methane production.