Evaluation of a single-flow continuous culture fermenter system for determination of ruminal fermentation and enteric methane production.

A 4-unit, single-flow continuous culture fermenter system was developed to assess in vitro nutrient digestibility, volatile fatty acid (VFA) concentration and daily enteric methane (CH4 ) production of ruminant diets. The objective was to develop a closed-vessel system that maintained protozoal populations and provided accurate predictions of total CH4 production. A diet of 50% orchardgrass (Dactylis glomerata L.) and 50% alfalfa (Medicago sativa L.) was fed during 4, 10-day periods (7-day adaptation and 3-day collection). Fermenters were fed 82 g of dry matter (DM)/day in four equal feedings. pH and temperature were taken every 2 min, and CH4 concentration was measured every 10 min. Samples for DM and protozoal counts were taken daily, and daily effluent samples were collected for determination of DM, VFA and NH3 -N concentrations. There was no effect (p > 0.17) of adaptation versus collection days on vessel and effluent DM, temperature or pH. Initial protozoal counts decreased (p < 0.01), but recovered to initial counts by the collection period. Total VFA, acetate, propionate and isobutyrate concentrations did not differ (p ≥ 0.13) among periods or days of the collection period. There was no difference (p ≥ 0.37) among days or periods in total daily CH4 production and CH4 production per g of OM, NDF, digestible OM or digestible NDF fed. Data collected throughout 4 experimental periods demonstrated that the system was able to reach a steady state in fermentation well within the 7-day adaptation period and even typically variable data (i.e., CH4 production) were stable within and across periods. While further research is needed to determine the relationship between this system and in vivo data, this continuous culture fermenter system provides a valid comparison of in vitro ruminal fermentation and enteric CH4 production of ruminant diets that can then be further validated with in vivo studies.

Enteric methane production and ruminal fermentation of forage brassica diets fed in continuous culture.

The aim of the current study was to determine nutrient digestibility, VFA production, N metabolism, and CH4 production of canola (Brassica napus L.), rapeseed (B. napus L.), turnip (Brassica rapa L.), and annual ryegrass (Lolium multiflorum Lam.) fed with orchardgrass (Dactylis glomerata L.) in continuous culture. Diets were randomly assigned to fermentors in a 4 × 4 Latin square design using 7 d for adaptation and 3 d for collection. Diets were: 1) 50% orchardgrass + 50% annual ryegrass (ARG); 2) 50% orchardgrass + 50% canola (CAN); 3) 50% orchardgrass + 50% rapeseed (RAP); and 4) 50% orchardgrass + 50% turnip (TUR). Feedings (82 g DM/d) occurred four times daily throughout 4, 10-d periods at 730, 1030, 1400, and 1900 h. Methane samples were collected every 10 min using a photoacoustic gas analyzer (LumaSense Technologies, Inc.; Santa Clara, CA) during the last 3 d of the experiment. Effluent samples were collected on d 8, 9, and 10, composited by fermentor, and analyzed for VFA and pH as well as DM, OM, CP, and fiber fractions for determination of nutrient digestibility. Forage samples were analyzed for CP, NDF, ADF, minerals, and glucosinolate (GLS) concentrations. Data were analyzed using the GLIMMIX procedure of SAS. Apparent DM, OM, and NDF digestibilities and true DM and OM digestibilities were similar (P > 0.28) among diets (45.1, 63.2, 44.1, 67.1, and 87.2%, respectively). Total VFA (87.2 mol/100 mol), pH (6.47), and acetate (A: 44.6 mol/100 mol) were also not different (P > 0.20) among diets. The A:P (P = propionate) ratio was greater (P < 0.01) in ARG and CAN than RAP and TUR. Daily CH4 production was greater (P < 0.01) in ARG than all other diets (68.9 vs. 11.2 mg/d). Methane, whether expressed as g per g of OM, NDF, digestible OM, or digestible NDF fed was greatest (P < 0.01) in ARG but similar (P > 0.18) among brassica diets. A significant negative correlation was observed between total GLS and CH4 production. However, when multiple regression analysis on CH4 production was completed, neither total GLS nor individual GLS were a significant component of the model. Addition of brassicas provided similar nutrient digestibility to ARG while reducing daily CH4 production, potentially making brassicas an alternative for ARG in pasture-based ruminant diets.