Fitting of pH conditions for the study of concentrate feeds fermentation by the in vitro gas-production technique

Abstract

Two experiments were conducted to simulate in vitro the fermentation conditions under high-concentrate feeding. The concentration of bicarbonate ion in the buffer of the incubation solution was assayed in Experiment 1, by adjusting medium pH to 6.50, 6.25, 6.00, 5.75 and 5.50, in two incubation series of 12 h, using barley as the reference substrate. The pH diminished linearly (P < 0001) by lowering the buffer, and remained constant throughout 12 h, except for treatments 5.75 and 5.50, where pH dropped to 5.51 and 5.31 at 12 h. Gas production decreased linearly with a decreasing medium pH (P < 0.001), with the total volume of gas produced after 12 h being highly dependent (P < 0.01) on pH at 12 h (R2 = 0.629), thus demonstrating the importance of the incubation pH for estimation of fermentation of concentrate feeds. In Experiment 2, the effect of pH on direct and indirect proportion of gas was studied by adding 0.0, 0.1, 0.2, 0.3, 0.4 and 0.5 mmol of acetic acid, either with or without (water added instead) rumen inoculum, to the media. Linear multiple regressions established between the volume of gas produced and the addition of acetic acid, and the bicarbonate ion concentration showed high determination coefficients for water (R2 = 0.929) and rumen inoculum (R2 = 0.851). Without inoculum, indirect gas production ranged from 9.4 to 12.4 mL/mmol of acid for medium pH of 5.50–6.50. With rumen inoculum, indirect gas was 20.8 mL/mmol acid, although this may have been biased by the contribution of inoculum itself to direct fermentation.