Effect of Clostridium butyricum Supplementation on in vitro Rumen Fermentation and Microbiota With High Grain Substrate Varying With Media pH Levels

Abstract

Clostridium butyricum (C. butyricum) can survive at low pH, and it has been widely used as an alternative to antibiotics for the improvement of feed efficiency and animal health in monogastrics. A recent study suggested that the improved ruminal fermentation with supplementing C. butyricum is may be associated with increasing the abundance of rumen microbiota in Holstein heifers, as ruminal pH plays a key role in rumen microbiota and the probiotics are often active in a dose-dependent manner. The objective of this study was to determine the effects of increasing the doses of C. butyricum on gas production (GP) kinetics, dry matter disappearance (DMD), fermentation characteristics, and rumen microbiota using a high grain substrate in batch culture varying with media pH levels. The doses of C. butyricum were supplemented at 0 (control), 0.5 × 106, 1 × 106, and 2 × 106 CFU/bottle, respectively, at either media pH 6.0 or pH 6.6. The fermentation microbiota at 0 and 1 × 106 CFU/bottle were determined using the 16S rRNA high throughput sequencing technology. Overall, the GP, DMD, total volatile fatty acid (VFA) concentration, and the ratio of acetate:propionate were higher (P <0.01) at media pH 6.6 than at pH 6.0. However, there was interaction between pH × dose of C. butyricum for rate constant of GP (P = 0.01), average GP rate (P = 0.07), and volume of GP (P = 0.06); with the increase in C. butyricum supplementation, the GP kinetics were not changed at media pH 6.0, but the volume (P = 0.02), rate of GP (P = 0.01), and average GP rate (P = 0.01) were quadratically changed at media pH 6.6. The DMD was not affected by increasing the supplementation of C. butyricum. The molar proportions of propionate (P <0.09), butyrate (P <0.06), and NH3-N concentration (P = 0.02) were quadratically changed with increasing supplementation of C. butyricum regardless of media pH levels. The interactions between media pH level and dose of C. butyricum supplementation were noticed for alpha diversity indexes of Shannon (P = 0.02) and Evenness (P = 0.04). The alpha diversity indexes increased (P <0.05) except for Chao1 with supplementation of C. butyricum. The unweighted uniFrac analysis showed that the group of control at media pH 6.0 and control at media pH 6.6, and supplementation of C. butyricum and control at media pH 6.0 clustered separately from each other. At the phylum level, relative abundance (RA) of Bacteroidota was lower (P <0.01) and Firmicutes was higher (P <0.01) at media pH 6.6 than pH 6.0. Moreover, RA of Proteobacteria decreased (P <0.05) with supplemented C. butyricum at either media pH 6.6 or pH 6.0. At media pH 6.6, RA of Rikenellaceae_RC9_gut_group and Prevotella were decreased, and CAG-352 was increased (at genus level) compared to pH 6.0. Supplementation of C. butyricum decreased RA of Rikenellaceae_RC9_gut_group and increased CAG-352 at media pH 6.0. It could hence be concluded that manipulating media pH level and supplementation of C. butyricum effectively modulated in vitro rumen fermentation characteristics and microbiota but in a dose depending manner of C. butyricum addition.

Effects of dietary Clostridium butyricum and rumen protected fat on meat quality, oxidative stability, and chemical composition of finishing goats

“…The trend of increase of HCW by CB is consistent with the increased LWBS. The increase in LWBS with CB in this study may be due to the fact that the CB can provide metabolites, especially short-chain FA, as an energy source to the digestive enzyme system, thus increase the BW [ 15 ]. The spleen is the largest immune organ in animal body, which can improve the body resistibility and minimize the pathogenic bacteria from invading the body, therefore its development is believed to be directly associated with the immune function of the body.…”

“…Moreover, butyric acid produced by CB metabolism can regulate muscle lipid metabolism [ 14 ]. Furthermore, the inconclusive effects of CB in the literature may be due to differences in source of CB strains used, its supplemental dose and type of diet [ 15 , 16 ]. Zhang et al [ 17 ] observed an interaction between dietary lipids and CB on lipid-related gene expression in breast muscle of broiler chickens, suggesting that dietary fat may be interacted with CB on meat quality.…”

Insights into Effects of Combined Capric and Lauric Acid on Rumen Bacterial Composition

“…An increased F:B ratio has been shown to contribute to improved bovine performance, including body weight gain and milk fat yield [ 29 ], but there are contradictory reports [ 30 ]. A greater abundance of Firmicutes indicates a more feed-efficient animal [ 21 , 31 ] with improved structural polysaccharide degradation [ 32 ]. Changes in this ratio did not affect digestibility, and we could not assess its effect on production parameters, because the animals in our study were dry cows [ 14 ].…”