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This study investigated the effect of saponins gypenoside (gynosaponins) on methane production and microbe numbers in a co-culture of a fungus, Piromyces sp. F1, and a methanogen, Methanobrevibacter sp.. Two co-culture systems were used: with methanogen (co-culture I) and without methanogen (co-culture II; methanogen growth inhibited by chloramphenicol). Each co-culture system was treated with 0, 50, 100 or 200 mg gynosaponins/l culture medium. Gas production, methane concentration and volatile fatty acid concentration (VFA) were measured for each treatment group. The numbers of anaerobic fungi and methanogen were quantified by real time PCR. The results showed that, compared with the control, gynosaponin significantly reduced the gas production, methane concentration, methane to TVFA ratio (total volatile fatty acid), TVFA concentration, number of fungi (except for 50 mg dose of gynosaponin in co-culture I) and number of methanogens. Methane was not detected in co-culture II. The individual VFAs proportion of TVFA were not affected by gynosaponins in either of the co-cultures. The pH was higher in both co-cultures than that of the control (P<0.01). These data suggest that gynosaponins has the potential for being used as feed additive to modulate the ruminal fermentation, inhibit the methanogen growth and reduce methane production.
Two trials were conducted to investigate the effect of a combination of essential oils (CEO) along with fumarate on in vitro rumen fermentation. In trial 1, the essential oil (EO) from thyme, oregano, cinnamon and lemon were mixed at five different ratios. The CEO were applied at levels of 0-500 mg/l. Addition of CEO decreased gas, methane, total volatile fatty acid (VFA) production at 24 h incubation in a dose-dependent manner. Methane tended to decrease much more than total VFA and gas at the same EO level. The mixture of oils at an equal ratio at 500 mg/l that decreased methane much more than VFA was chosen as the optimal combination. In trial 2, the optimal combination was used with 0, 5, 10 or 15 mmol/l of monosodium fumarate. Addition of fumarate further decreased methane production, with 10 mmol/l fumarate resulting in the largest reduction (80.2%) and the smallest decrease in total VFA (5.7%) and gas production (16.7%). Quantification of several ruminal microbe populations by RT-PCR showed that the optimal combination sharply decreased ruminal protozoa; the populations of fungi and fibrolytic bacteria were also decreased. In summary, at an appropriate level, CEO can inhibit methane production. Inclusion of fumarate can further decrease it, which is attributed mainly to inhibition of protozoa and methanogens.
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