Differences in yields of microbial crude protein from in vitro fermentation of carbohydrates

The yield of microbial crude protein (CP) from carbohydrate fermentations was examined using trichloroacetic acid (TCA) precipitation of batch cultures. The medium contained ammonium bicarbonate, casein acid hydrolysate, and cysteine hydrochloride as nitrogen sources. Isolated bermudagrass neutral detergent fiber (iNDF) and 60:40 blends of iNDF and sucrose (Sue), citrus pectin (Pee), or corn starch (Sta) (similar to 375 mg of substrate organic matter/vial) were fermented in vitro in two separate fermentation runs with mixed ruminal microbes. Three fermentation tubes for each substrate were destructively sampled at 0, 4, 8, 12, 16, 20, and 24 h. Fermented samples were precipitated at a concentration of 19.4% TCA, and filtered to collect unfermented iNDF and precipitate. Collected residues were analyzed for CP as Kjeldahl N x 6.25. Microbial CP (TCACP) was estimated as TCA-precipitated CP corrected for the TCA-precipitated CP content of substrates at 0 h, and the mean of fermentation blanks from each hour. Medium pH did not decline below 6.49 in any fermentation tube. Comparisons of maximal yields based on the hour in which the measured mean yield was greatest for each substrate in each fermentation indicated that Sta > Sue = Pee > iNDF (P < 0.05). All substrates showed increases in TCACP to their maxima, followed by declines in TCACP. This likely reflects the relative dominance of production or degradation of microbes about the point of substrate limitation. Unlike other substrates, Sue had no detectable lag, and presented a more persistent TCACP yield curve than the other non-NDF carbohydrates (NFC). Regression analysis of TCACP yield over time for iNDF versus other substrates, Pee + Sta versus Sue, and Pee versus Sta indicated that the compared curves were not parallel (P < 0.05). The patterns of TCACP yield over time were cubic for iNDF and Sue, and quartic for Pec and Sta. The maximal yields of TCACP predicted from the regressions were Sta: 34.0 mg at 15.6 h, Pee: 29.9 mg at 13.5 h, Sue: 25.5 mg at 12.6 h, and iNDF: 13.6 mg at 19.3 h. The NDF and NFC carbohydrates examined differed in both maximal yields and temporal patterns of yield of TCACP.