In vitro rumen incubation studies were conducted to determine effects of initial pH on bacterial attachment and fiber digestion. Ruminal fluid pH was adjusted to 5.7, 6.2 and 6.7, and three major fibrolytic bacteria attached to rice straw in the mixed culture were quantified with real-time PCR. The numbers of attached and unattached Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminocococcus albus were lower (p<0.05) at initial pH of 5.7 without significant difference between those at higher initial pH. Lowering incubation media pH to 5.7 also increased bacterial numbers detached from substrate regardless of bacterial species. Dry matter digestibility, gas accumulation and total VFA production were pH-dependent. Unlike bacterial attachment, maintaining an initial pH of 6.7 increased digestion over initial pH of 6.2. After 48 h in vitro rumen fermentation, average increases in DM digestion, gas accumulation, and total VFA production at initial pH of 6.2 and 6.7 were 2.8 and 4.4, 2.0 and 3.0, and 1.2 and 1.6 times those at initial pH of 5.7, respectively. The lag time to reach above 2% DM digestibility at low initial pH was taken more times (8 h) than at high and middle initial pH (4 h). Current data clearly indicate that ruminal pH is one of the important determinants of fiber digestion, which is modulated via the effect on bacterial attachment to fiber substrates.
A series of in vitro and in vivo experiments were conducted to investigate the effects of the mixture of Tween 80 and cellulolytic enzymes (xylanase and cellulase) on total tract nutrient digestibility and rumen cellulolytic bacterial adhesion rates in Holstein steers. Ground timothy hay sprayed with various levels of Tween 80 and cellulolytic enzymes was used as substrates in an in vitro experiment to find out the best combinations for DM degradation. The application level of 2.5% (v/w) Tween 80 and the combination of 5 U xylanase and 2.5 U cellulase per gram of ground timothy hay (DM basis) resulted in the highest in vitro dry matter degradation rate (p<0.05). Feeding the same timothy hay to Holstein steers also improved in vivo nutrient (DM, CP, CF, NDF and ADF) digesibilities compared to non-treated hay (p<0.05). Moreover, Tween 80 and enzyme combination treatment increased total ruminal VFA and concentrations of propionic acid and isovaleric acid with decreased acetate to propionate ratio (p<0.001). However, adhesion rates of Fibrobacter succinogenes and Ruminococcus flavefaciens determined by Real Time PCR were not influenced by the treatment while that of Ruminococcus albus was decreased (p<0.05). The present results indicate that a mixture of Tween 80 and cellulolytic enzymes can improve rumen environment and feed digestibility with variable influence on cellulolytic bacterial adhesion on feed.
This study was conducted to investigate the effect of biological membrane transfer modifier, lysophospholipd (LPLs) on the parameters from in vitro rumen simulated fermentation. Commercially available LPLs product (Lipidol TM ) was supplemented into experimental diets which consisted of orchard grass and concentrate diet (60:40) in different levels (0.1%, 0.3% and 0.5%). Then in vitro rumen simulated fermentation was performed. Although, a declining trend of pH was found in treatments, all pH values were detected in a range relevant to normal rumen fermentation. Gas production, ammonia nitrogen and total VFA production were greatly influenced by the supplementation of LPLs. All parameters were increased along with increased levels of LPLs in diet. As a result, 0.1% of Lipidol TM is recommended based on the determined in vitro rumen fermentative parameters in this study.
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