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The purpose of this experiment was to study the effects of different levels of Clostridium butyricum supplementation on the gas production, fermentation parameters, and nutrient digestibility in TMR diet by in vitro gas production method, so as to determine the appropriate amount of Clostridium butyricum supplementation in beef cattle diet. Holstein steers with permanent fistula were selected as rumen fluid donors and TMR as culture substrate. The control group (CON group) only contained the culture substrate, while the Clostridium butyricum test group added Clostridium butyricum with the concentrations of 2x108 (CB1 group), 4x108 (CB2 group), 6x108 (CB3 group), 8x108 (CB4 group) and 10x108 CFU/kg (CB5 group) on the basis of the culture substrate, with six replicates at each level. The results showed that the in vitro gas production of CB1 group was significantly lower than that of other groups when the fermentation time was 2 h and 4 h (P<0.05). When the fermentation time was 12 h, the in vitro gas production of CB3 group, CB2 group, CB4 group, and CB5 group was significantly higher than that of CON group and CB1 group (P<0.05). The pH value in each experimental group were significantly lower than those in CON group (P<0.05), while the content of ammonia nitrogen (NH3-N) was significantly higher than that in CON group (P<0.05), and there was no significant difference among the experimental groups (P>0.05). The concentrations of acetic acid, propionic acid, and total acid in CB3 group, CB4 group and CB5 group were significantly higher than those in CON group (P<0.05). Isobutyric acid content in CB1 group, CB2 group and CB3 group was significantly higher than that in CON group (P<0.05). Dry matter digestibility (DMD) and neutral detergent fiber degradation (NDFD) of each experimental group were significantly higher than those of CON group (P<0.05). The degradation rate of acid detergent fiber (ADFD) in CB2 group was significantly higher than that in CON group (P<0.05). The study indicates that the addition of Clostridium butyricum can increase gas production, NH3-N, acetic acid, propionic acid, total volatile acid content and DMD. The suitable addition of Clostridium butyricum is 6x108 CFU/kg. [ABSTRACT FROM AUTHOR]

Highlights • The Moringa oleifera leaves and peduncles are a new feed resource. • The replacement of alfalfa hay by M. oleifera improved rumen fermentation. • The M. oleifera increased intake and apparent digestibility. Abstract To study the effects of replacing alfalfa hay with Moringa oleifera leaves and peduncles in diets on intake, digestibility, and rumen fermentation in dairy cows, eight multiparous Holsteins cows with similar DIM (day in milk, 93 ± 18 d) and body weights (624 ± 38 kg) were used in a crossover design with two dietary treatments. The treatment diets contained similar concentrate mixtures and corn silage but different proportions of M. oleifera leaves and peduncles. The treatments were a (1) control diet (0% M. oleifera feed) and (2) M. oleifera diet (10.85% M. oleifera feed). The M. oleifera feed used M. oleifera leaves and M. oleifera peduncles at a ratio of 3:2 and replaced portions of alfalfa hay. The experimental period was 18 d, with 15 d for treatment adaptation and 3 d for data collection. Compared with the control diet, the dry matter intake and apparent digestibility of dry matter, crude protein, acid detergent fiber and neutral detergent fiber in the M. oleifera diet were significantly increased. No difference was observed in ruminal pH; however, the ammonia-N concentrate was lower for the M. oleifera diet than the control diet. The concentrations of total ruminal volatile fatty acids, acetate, propionate, and butyrate were higher in cows fed the M. oleifera diet than in those fed the control diet. The results of ruminal microbes showed that those consuming the M. oleifera diet had greater relative expression level of Fibrobacter succinogenes, Butyrivibrio fibrisolvens, Ruminobacter amylophilus, Streptococcus bovis, Prevotella ruminicola which were observed for the control diet than the M. oleifera diet. These results show that the M. oleifera diet can improve intake and apparent digestibility of nutrients, and rumen fermentation in dairy cows. [ABSTRACT FROM AUTHOR]