Cottonseed meal (CSM) is an important protein feed source for dairy cows. Its inclusion in ruminant diets is limited due to the presence of the highly toxic gossypol though rumen microorganisms are believed to be capable of gossypol degrading and transforming. The objective of the present study was to isolate the gossypol-degrading bacteria from the rumen contents and to assess its potential for gossypol degradation in vitro. A strain named Lactobacillus agilis WWK129 was anaerobically isolated from dairy cows after mixed rumen microorganisms were grown on a substrate with gossypol as the sole carbon source. Furthermore, the strain was applied at 5% inoculum concentration in vitro to continuously ferment CSM at 39 °C for five days, and it presented gossypol degradability as high as 83%. Meanwhile, the CSM contents of crude protein, essential amino acids increased significantly along with the increase of lactic acid yield (p < 0.01). Compared with the original CSM, the fermented CSM contents of neutral detergent fiber and acid detergent fiber was remarkably decreased after the anaerobic fermentation (p < 0.01). In brief, the Lactobacillus strain isolated from the rumen is not only of great importance for gossypol biodegradation of CSM, but it could also be used to further explore the role of rumen microorganisms in gossypol degradation by the ruminants.

Cysteamine (CS) is an essential nutritional regulator that improves the productive performance of animals by regulating somatotropic hormone secretion. To investigate the fattening potential and effects of CS on rumen microbial fermentation, 48 feedlot lambs were randomly assigned to four groups and fed diets supplemented with different CS concentrations (0, 20, 40, and 60 mg/kg BW). An increase in dietary CS concentrations linearly increased the average daily gain (ADG) and dry matter intake (p < 0.05) but decreased the feed-to-gain ratio (p < 0.01). For the serum hormone, increasing the dietary CS concentration linearly decreased somatostatin and leptin concentration (p < 0.01) but linearly increased the concentration of growth hormone and insulin-like growth factor 1 (p < 0.01). Regarding rumen fermentation, ruminal pH, ammonia-N, and butyrate content did not differ among the four treatments, although dietary CS supplementation linearly increased microbial protein and propionate and decreased the amount of acetate (p < 0.05). Furthermore, an increase in dietary CS concentrations quadratically decreased the estimated methane production and methane production per kg ADG (p < 0.05). High-throughput sequencing revealed that increased dietary CS concentrations quadratically increased Prevotella (p < 0.05), and Prevotella and norank_f__norank_o__Clostridia_UCG-014 were positively correlated with growth performance and rumen fermentation in a Spearman correlation analysis (r > 0.55, p < 0.05). Overall, a CS concentration higher than 20 mg/kg BW produced growth-promoting effects by inhibiting somatostatin concentrations and shifting the rumen toward glucogenic propionate fermentation by enriching Prevotella. In addition, Prevotella and norank_f__norank_o__Clostridia_UCG-014 were positively correlated with growth performance in lambs.

Corn silage is an important source of forage, but whether or not bacterial inoculants should be applied is somewhat controversial in ruminant feeding practice. In the present study, chopped whole corn plants treated with a single inoculant of Lactobacillus buchneri (LB), Lactobacillus plantarum (LP), Pediococcus pentosaceus (PP) served as either homofermentation (e.g., lactate only) or heterofermentation (e.g., lactate and acetate) controls and compared with those treated with either a mixture of the lactic acid bacteria (QA: 60% LP, 10%PP, 30% LB) or a mixture of the lactic acid bacteria (QB: 60% LP, 15% PP, 25% LB), to investigate their effects on the fermentation quality, ester-linked phenolic acids, and in vitro digestibility. After 60 day ensiling, the addition of QA exhibited the lowest pH (3.51) with greater lactic acid (LA) production. The ester-linked ferulic acid (FAest) and p-coumaric acid (pCAest) concentrations were significantly decreased during 60 days ensiling. And among all these groups, the LB and QA treated group showed a lower concentration of FAest and pCAest than other groups. After 60 days ensiling, Lactobacillus was the dominant genus in all LAB treated groups. Meanwhile, negative correlations of Bacillus, Bacteroides, Bifidobacterium, Blautia, Prevotella, Ruminococcus, and Roseburia with FAest content after 60 days ensiling occurred in the present study. Komagataeibacter was mainly found in LB and PP addition silages, and presented a significant negative effect with the level of acid detergent fiber (ADF). To explore whether the addition of LABs can improve digestibility of whole corn silage, an in vitro rumen fermentation was conducted using the 60 day ensiled whole corn silages as substrates. The QA addition group exhibited a greater 48 h and 96 h in vitro dry matter and ADF disappearance, greater 48 h gas production and less methane emissions. Even though there were the same neutral NDF levels in corn silages treated with LB and QA after 60 days ensiling, the QA treated silages with lower FAest and pCAest presented higher IVDMD after 96 h and 48 h in vitro fermentation. In brief, the addition of mixed inoculants of 60% LB,10% PP, 30% LB compared with the addition of whichever single HoLAB or HeLAB inoculants, facilitated the release of ester-linked phenolic acids (e.g., ferulic and p-coumaric acids) and remarkably, improved silage quality in terms of sharp pH decline and greater lactate production. Taken together with the improvement in rumen microbial fermentation, the results obtained in the present study provided concrete evidence for the role of mixed LAB application in corn silage preparation for ruminant feeding practices.

Cysteamine (CS) is a vital antioxidant product and nutritional regulator that improves the productive performance of animals. A 2 × 4 factorial in vitro experiment was performed to determine the effect of the CS supplementation levels of 0, 20, 40, and 60 mg/g, based on substrate weight, on the ruminal fermentation, antioxidant capacity, and microorganisms of a high-forage substrate (HF, forage:corn meal = 7:3) in the Statistical Analysis System Institute. After 48 h of incubation, the in vitro dry matter disappearance and gas production in the LF group were higher when compared with a low-forage substrate (LF, forge hay:corn meal = 3:7), which was analyzed via the use of the MIXED procedure of the HF group, and these increased linearly with the increasing CS supplementation (p < 0.01). With regard to rumen fermentation, the pH and acetate were lower in the LF group compared to the HF group (p < 0.01). However, the ammonia N, microbial crude protein, total volatile fatty acids (VFA), and propionate in the LF group were greater than those in the HF group (p < 0.05). With the CS supplementation increasing, the pH, ammonia N, acetate, and A:P decreased linearly, while the microbial crude protein, total VFA, and propionate increased linearly (p < 0.01). Greater antioxidant capacity was observed in the LF group, and the increasing CS supplementation linearly increased the superoxide dismutase, catalase, glutathione peroxidase, total antioxidant capacity, glutathione, and glutathione reductase, while it decreased the malondialdehyde (p < 0.05). No difference occurred in the ruminal bacteria alpha diversity with the increasing CS supplementation, but it was higher in the LF group than in the HF group (p < 0.01). Based on the rumen bacterial community, a higher proportion of Bacteroidota, instead of Firmicutes, was in the LF group than in the HF group. Furthermore, increasing the CS supplementation linearly increased the relative abundance of Prevotella, norank_f_F082, and Prevotellaceae_UCG-001 under the two substrates (p < 0.05). Prevotella, norank_f_F082, and Prevotellaceae_UCG-001 were positively correlated with gas production, rumen fermentation, and antioxidant capacity in a Spearman correlation analysis (r > 0.31, p < 0.05). Overall, a CS supplementation of not less than 20 mg/g based on substrate weight enhanced the rumen fermentation and rumen antioxidant capacity of the fermentation system, and it guided the rumen fermentation towards glucogenic propionate by enriching the Prevotella in Bacteroidetes.

Guanidine acetic acid (GAA) has been reported to improve growth performance, nutrient utilization, and meat quality in livestock. This study aimed to investigate whether coated GAA (CGAA) in comparison with uncoated GAA (UGAA) could have different effects on rumen fermentation, antioxidant capacity, and microflora composition in the rumen. Seventy-two lambs were randomly arranged in a 2 × 3 factorial experiment design with two diets of different forage type (OH: oaten hay; OHWS: oaten hay plus wheat silage) and three GAA treatments within each diet (control, diet without GAA addition; UGAA, uncoated GAA; CGAA, coated GAA). The whole feeding trial lasted for 120 days. The lambs in the OH group presented lower total volatile fatty acid (VFA), alpha diversity, Firmicutes, NK4A214_group, and Lachnospiraceae_NK3A20_group than those on the OHWS diet in the last 60 days of the feeding stage (p < 0.05). Regardless of what GAA form was added, dietary GAA supplementation increased the total VFA, microbial crude protein (MCP), adenosine triphosphate (ATP), and antioxidant capacity in rumen during lamb feedlotting (p < 0.05). However, molar propionate proportion, acetate:propionate ratio (A:P), and relative Succiniclasticum abundance decreased with GAA addition in the first 60 days of the growing stage, while the molar butyrate proportion and NK4A214_group (p < 0.05) in response to GAA addition increased in the last 60 days of feeding. These findings indicated that dietary GAA enhanced antioxidant capacity and fermentation characteristics in the rumen, but the addition of uncoated GAA in diets might cause some dysbacteriosis of the rumen microbiota.

Gossypol, a phenolic compound found in the cotton plant, is widely distributed in cottonseed by-products. Although ruminant animals are believed to be more tolerant of gossypol toxicity than monogastric animals due to rumen microbial fermentation, the actual mechanisms of detoxification remain unclear. In contrast, the metabolic detoxification of gossypol by Helicoverpa armigera (Lepidoptera: Noctuidae) larvae has achieved great advances. The present review discusses the clinical signs of gossypol in ruminant animals, as well as summarizing advances in the study of gossypol detoxification in the rumen. It also examines the regulatory roles of several key enzymes in gossypol detoxification and transformation known in H. armigera. With the rapid development of modern molecular biotechnology and -omics technology strategies, evidence increasingly indicates that research into the biological degradation of gossypol in H. armigera larvae and some microbes, in terms of these key enzymes, could provide scientific insights that would underpin future work on microbial gossypol detoxification in the rumen, with the ultimate aim of further alleviating gossypol toxicity in ruminant animals.