In Vitro Fermentation Vessel Type and Method Alter Fiber Digestibility Estimates

Abstract

Selected vessel types and conditions used for in vitro fermentation were compared to evaluate their effects on determinations of neutral detergent fiber (NDF) digestibility (NDFD) in 2 replicate 48-h fermentations. Treatments included 50-mL polyethylene centrifuge tubes with gas-release valves (treatment 1); 50-mL polyethylene centrifuge tubes with continuous gassing with CO2 (treatment 2); 50-mL polyethylene centrifuge tubes sealed, oriented horizontally, and shaken continuously parallel to the long axis of the tube, with manual gas release (treatment 3); 125-mL Erlenmeyer flasks with continuous gassing with CO2 (treatment 4); and 125-mL serum vials sealed with stoppers and crimp seals with (treatment 5) or without (treatment 6) manual gas release. Goering and Van Soest medium and blended ruminal inoculum from 4 lactating cows were used. Substrates were alfalfa hay, corn silage, ryegrass hay, and soyhulls. Gas was released and measured in treatments 3 and 6 at 3.0, 5.5, 9.0, 11.5, 23.5, 29.5, and 47.5h by using a syringe with a hypodermic needle. Vessels from each treatment were harvested at 0, 6, 12, 24, 30, and 48h for NDF analysis, with NDFD calculated as 1-[(residual NDF, g-residual NDF in fermentation blank, g)/sample NDF, g]. Medium pH did not decline below 6.3 for any treatment. Average values for NDFD for 24 through 48h were 0.576, 0.639, 0.688, 0.668, 0.679, and 0.681 for treatments 1 through 6, respectively (standard error of the difference=0.008). The lowest NDFD was noted for treatment 1, which differed from all other treatments; treatments 3, 4, 5, and 6 did not differ by treatment or by the interaction of treatment and substrate. Treatments 1 and 2 gave lower NDFD values than the other treatments, but these differences were not consistent and differed by substrate, with alfalfa showing the fewest differences among treatments and soyhulls the most. Net energy of lactation values for substrates, as predicted from differences in 48-h NDFD, were 7 to 15% lower for treatment 1 than for the average of all other treatments. Slopes of the gas production per gram of substrate dry matter curves differed between treatments 3 and 5. In conclusion, measured NDFD was altered by fermentation treatment, with polyethylene tubes + gas-release valves giving the lowest values. Consequently, NDFD values may not be comparable across fermentation methods, but the effect will vary among feedstuffs. The combination of methods used for sealing, gassing, or agitating vessels may have a greater impact on NDFD than does vessel type.