Comparison of in vivo and in vitro techniques for methane production from ruminant diets

This study was conducted to compare the methane (CH4) production estimated by in vivo (sulfur hexafluoride tracer technique (SF6)) with that of two in vitro rumen simulation (RUSITEC) and gas production (IVGPT)) techniques. Four adult dry Holstein cows, aged 7.4 +/- 3.0 years and weighing 697 +/- 70 kg, were used for measuring methane production from five diets by the SF6 technique. The experimental diets were alfalfa hay (D-1), corn silage + soybean meal (SBM) (910: 90, D-2), Italian rye grass hay +SBM (920: 80, D-3) rice straw +SBM (910: 90, D-4) and Sudan grass hay +SBM (920: 80, D-5). Each diet was individually fed to all 4 cows and 5 feeding studies of 17 d each were conducted to measure the methane production. In the RUSITEC, methane production was measured from triplicate vessels for each diet In vitro gas production was measured for each of the diets in triplicate syringes. The gas produced after 24 and 48 h was recorded and gas samples were collected in vacuum vials and the methane production was calculated after correction for standard temperature and pressure (STP). Compared to the SF6 technique, estimates of methane production using the RUSITEC were lower for all diets. Methane production estimated from 24 h in vitro gas production was higher (p < 0.001) on D-1 as compared to that measured by SF6, whereas on D2 to D5 it was lower. Compared to SF6, methane production estimated from 48 h in vitro gas production was higher on all diets. However, methane estimated from the mean of the two measurement intervals (24+48 h/2) in IVGPT was very close to that of SF6 (correlation 0.98), except on D-1. The results of our study confirmed that IVGPT is reflective of in vivo conditions, so that it could be used to generate a database on methane production potential of various ruminant diets and to examine strategies to modify methane emissions by ruminants.