The effect of pectin, corn and wheat starch, inulin and pH on in vitro production of methane, short chain fatty acids and on the microbial community composition in rumen fluid

Methane emission from livestock, ruminants in particular, contributes to the build up of greenhouse gases in the atmosphere. Therefore the focus on methane emission from ruminants has increased. The objective of this study was to investigate mechanisms for methanogenesis in a rumen fluid-based in vitro fermentation system as a consequence of carbohydrate source (pectin, wheat and corn starch and inulin) and pH (ranging from 5.5 to 7.0). Effects were evaluated with respect to methane and short chain fatty acid (SCFA) production, and changes in the microbial community in the ruminal fluid as assessed by terminal-restriction fragment length polymorphism (T-RFLP) analysis. Fermentation of pectin resulted in significantly lower methane production rates during the first 10 h of fermentation compared to the other substrates (P = 0.001), although total methane production was unaffected by carbohydrate source (P = 0.531). Total acetic acid production was highest for pectin and lowest for inulin (P < 0.001) and vice versa for butyric acid production from pectin and inulin (P < 0.001). Total propionic acid production was unaffected by the carbohydrate source (P = 0.791). Methane production rates were significantly lower for fermentations at pH 5.5 and 7.0 (P = 0.005), sustained as a trend after 48 h (P = 0.059), indicating that there was a general optimum for methanogenic activity in the pH range from 6.0 to 6.5. Decreasing pH from 7.0 to 5.5 significantly favored total butyric acid production (P < 0.001). Principle component analysis of T-RFLP patterns revealed that both pectin and pH 5.5 resulted in pronounced changes in the microbial community composition. This study demonstrates that both carbohydrate source and pH affect methane and SCFA production patterns, and the microbial community composition in rumen fluid.

► We investigated mechanisms of methanogenesis in rumen fluid.
► Substrates were pectin, inulin, wheat and corn starch fermented at pH 5.5, 6, 6.5 and 7.
► Total CH4 production was highest at pH 6 and 6.5 and lowest for pectin fermentation.
► CH4 production was less affected by carbohydrate source than pH.
► The bacterial community composition was particularly affected by pectin and pH 5.5.