Enteric methane (CH4) production is a side-effect of herbivore digestion, but it is unknown whether CH4 itself influences digestive physiology. We investigated the effect of adding CH4 to, or reducing it in, the reticulorumen (RR) in a 4×4 Latin square experiment with rumen-fistulated, non-lactating cows, with four treatments: (i) control, (ii) insufflation of CH4 (iCH4), (iii) N via rumen fistula, (iv) reduction of CH4 via administration of bromochloromethane (BCM). DM intake (DMI), apparent total tract digestibility, digesta mean retention times (MRT), rumen motility and chewing activity, spot breath CH4 emission (CH4exhal, litre/kg DMI) as well as CH4 dissolved in rumen fluid (CH4RRf, µg/ml) were measured. Data were analysed using mixed models, including treatment (or, alternatively, CH4exhal or CH4RRf) and DMI relative to body mass0·85 (rDMI) as covariates. rDMI was the lowest on the BCM treatment. CH4exhal was highest for iCH4 and lowest for BCM treatments, whereas only BCM affected (reduced) CH4RRf. After adjusting for rDMI, CH4RRf had a negative association with MRT in the gastrointestinal tract but not in the RR, and negative associations with fibre digestibility and measures of rumination activity. Adjusting for rDMI, CH4exhal had additionally a negative association with particle MRT in the RR and a positive association with rumen motility. Thus, higher rumen levels of CH4 (CH4exhal or CH4RRf) were associated with shorter MRT and increased motility. These findings are tentatively interpreted as a feedback mechanism in the ruminant digestive tract that aims at mitigating CH4 losses by shortening MRT at higher CH4.

The effects of the anti-methanogenic compound, bromochloromethane (BCM), on rumen microbial fermentation and ecology were examined in vivo. Japanese goats were fed a diet of 50 % Timothy grass and 50 % concentrate and then sequentially adapted to low, mid and high doses of BCM. The goats were placed into the respiration chambers for analysis of rumen microbial function and methane and H2 production. The levels of methane production were reduced by 5, 71 and 91 %, and H2 production was estimated at 545, 2941 and 3496 mmol/head per d, in response to low, mid and high doses of BCM, respectively, with no effect on maintenance feed intake and digestibility. Real-time PCR quantification of microbial groups showed a significant decrease relative to controls in abundance of methanogens and rumen fungi, whereas there were increases in Prevotella spp. and Fibrobacter succinogenes, a decrease in Ruminococcus albus and R. flavefaciens was unchanged. The numbers of protozoa were also unaffected. Denaturing gradient gel electrophoresis and quantitative PCR analysis revealed that several Prevotella spp. were the bacteria that increased most in response to BCM treatment. It is concluded that the methane-inhibited rumen adapts to high hydrogen levels by shifting fermentation to propionate via Prevotella spp., but the majority of metabolic hydrogen is expelled as H2 gas.

Bromochloromethane (BCM), a halogenated methane analogue, was evaluated for its anti-methanogenic activity in both batch and continuous fermentation systems. For batch fermentation, a roughage-based substrate was incubated with mixed rumen microflora for 24 h at two concentrations of BCM (5 and 10 μm). A 89–94 % reduction in methane was obtained in terms of volume and truly degraded substrate (TDS; P < 0·05). The partitioning factor (an index of efficiency of microbial protein production; expressed as mg TDS/ml net gas produced) increased from 3·55 to 3·73 (P < 0·05), while the acetate:propionate proportion decreased from 2·80 to 2·22 (P < 0·05). A complete inhibition of methanogens was associated with a 48 % decrease in Ruminococcus flavefaciens, a 68 % increase in Fibrobacter succinogenes and a 30 % increase in rumen fungi when quantified using qPCR. The continuous fermentation was carried out using the roughage-based substrate in four fermenters, two fermenters being control and the other two fed with BCM (5 μm) once in a day, for nine consecutive days. A persistent effect of BCM on methane reduction (85–90 %) was obtained throughout the study (P < 0·05) with no effect on gas production, SCFA production, acetate:propionate proportion, true degradability and efficiency of microbial mass synthesis (P>0·05). The complete inhibition of methane resulted in a significant decrease in R. flavefaciens and methanogens (P < 0·05) and an increase in fungal population (P < 0·05), while there was no effect on total bacterial and F. succinogenes populations (P>0·05). The batch fermentation confirms the anti-methanogenic activity of BCM, while the continuous fermentation indicates the persistency of this effect under in vitro conditions.