The effect of sucrose supplements on particle-associated carboxymethylcellulase (EC 3.2.1.4) and xylanase (EC 3.2.1.8) activities in cattle given grass-silage-based diet

Abstract

Carboxymethylcellulase (EC 3 . 2 . 1 . 4 ; CMCase) and xylanase (EC 3 . 2 . 1 .8) activities were assayed in rumen fluid and from microbes closely associated either with rumen particulate material or with feed particles incubated in nylon bags in the rumen of cattle. The cattle were fitted with a permanent rumen cannula and a simple 'T'-piece duodenal cannula and were given four diets in a 4 x 4 Latin Square experiment. The basal diet (diet C) consisted of grass silage, barley and rapeseed meal (700, 240 and 60 g/kg total dry matter (DM)) given at the rate of 5.3 kg/d or supplemented with 1.0 kg sucroseld given twice daily (diet S), twice daily with 0.25 kg sodium bicarbonate/d (diet B) or as a continuous intrarumen infusion (diet I). Giving sucrose supplements decreased CMCase and xylanase activities extracted from microbes associated with rumen particulate material or feed particles incubated in nylon bags as compared with diet C. Supplementation of the sucrose diet with sodium bicarbonate resulted in higher CMCase and xylanase activities than other sucrose diets (S and I). Particle-associated CMCase and xylanase activities were found to be very sensitive in detecting differences in the rumen environment and were related to changes in cell wall digestion. The activities were highly correlated with disappearance of DM and neutral-detergent fibre from nylon bags incubated in the rumen, rumen and total digestion of cell-wall carbohydrates and rumen pool size of cell-wall carbohydrates. It was concluded that the attachment of fibrinolytic enzymes is involved in the depression of fibre digestion. Particle-associated CMCase and xylanase activities were much higher when measured from rumen particulate material than from feed particles incubated in nylon bags.

Supplementation with non-fibrous carbohydrates reduced fiber digestibility and did not improve microbial protein synthesis in sheep fed fresh forage of two nutritive values

“…Calsamiglia et al (2008), working in vitro and Martin et al (1999) working in vivo, attributed the reduction in NDF digestibility primarily to pH changes. Moreover, Huhtanen and Khalili (1992), working in vivo, reported that the use of supplementation of diets containing sucrose with sodium bicarbonate, resulted in higher carboxymethylcelulase and xylanase activities than sucrose diets without buffer. Martin and Michalet-Doreau (1995), working with barley-supplemented cows with or without infusions of bicarbonate salts observed that, although the density of microorganisms was similar, glycosidase activities were higher when buffer was added.…”

Effect of extraction method on activities of polysaccharide-depolymerase enzymes in the microbial population from the solid phase in the rumen

“…The grinding improves microbial accessibility while freezing/defrosting improves rupture of microbial cells. The Gram-negative bacteria, which are predominant in the rumen, are considered to be particularly susceptible to this treatment (Hammond, 1992 Huhtanen and Khalili, 1992); and diminution of activities after feeding (Williams ef al, 1989;Bowman and Firkins, 1993;Martin et al, 1993 …”

“…This measurement requires the extraction of these enzymes from microbes adhering to rumen content. Extractions by sonication (Williams and Strachan, 1984;Martin et al, 1993) or lysozyme and carbon tetrachloride (CC' 4 ) treatment (Silva ef al, 1987;Huhtanen and Khalili, 1992;Bowman and Firkins, 1993) (Williams et al, 1989) and dietary composition (Huhtanen and Khalili, 1992).…”

The effect of live yeast, <i>Saccharomyces cerevisiae</i>, and their metabolites on ciliate fauna, fibrolytic and amylolytic activity, carbohydrate digestion and fermentation in the rumen of goats

“…Enzymes were extracted from the rumen content with CCl 4 (Huhtanen and Khalili, 1992) and the activities carboxymethylocellulase (CMC-ase), xylanase, and amylase were determined according to Michałowski et al (2003) and Bełżecki et al (2007) using the dinitrosalicilic acid reagent. The VFA concentration was determined by gas chromatography according to Ziołecki and Kwiatkowska (1973).…”