Methodological variations in the amount of sample incubated and the type of rumen fluid used are commonly observed. This study evaluated the effect of three sample amounts (0.6, 1.3 or 2.6g DM 100mL-1 of rumen fluid) incubated in rumen fluid buffered (BRF-buffered rumen fluid) or not (PRF-pure rumen fluid) on total gas volume (TV), methane (CH4) production, dry matter degradation (DM Deg) and final pH of corn silage incubated in vitro. The highest DM Deg was reached with the lowest amounts of sample (0.6 and 1.3g of DM 100mL-1 of rumen fluid). The sample amount of 2.6g associated with PRF reduced CH4 production (P<0.05), compared with the amounts of 0.6 and 1.3g DM 100mL-1, which had similar CH4 production (P>0.05). The use of BRF caused no effect on CH4 production (P>0.05), independent of the sample amount. Increasing the amount of substrate resulted in lower final pH of incubation in both fluids (P<0.05). Our results indicate that incubations should be performed with the smallest amount of sample (0.6g of DM 100mL-1 of rumen fluid), using fluid without buffer. Incubation without buffer solution overestimates the CH4 production of corn silage. Further studies should be conducted to verify the possibility of in vitro ruminal incubation of other ingredients using pure rumen fluid.
degradation; gas; incubation; methane; pH
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