Influence of protein fermentation and carbohydrate source on in vitro methane production.

Incubations were carried out with batch cultures of ruminal micro-organisms from sheep to analyse the influence of the N source on in vitro CH4 production. The two substrates were mixtures of maize starch and cellulose in proportions of 75:25 and 25:75 (STAR and CEL substrates, respectively), and the three nitrogen (N) sources were ammonia (NH4 Cl), casein (CA) and isolated soya bean protein (SP). Five isonitrogenous treatments were made by replacing non-protein-N (NPN) with CA or SP at levels of 0 (NPN), 50 (CA50 and SP50, respectively) and 100% (CA100 and SP100) of total N. All N treatments were applied at a rate of 35 mg of N/g of substrate organic matter and incubations lasted 16.5 h. With both proteins, N source × substrate interactions (p = 0.065 to 0.002) were detected for CH4 production and CH4 /total VFA ratio. The increases in CH4 production observed by replacing the NPN with protein-N were higher (p < 0.05) for STAR than for CEL substrate, but the opposite was observed for the increases in volatile fatty acid (VFA) production. As a consequence, replacing the NPN by increased levels of CA or SP led to linear increases (p < 0.05) in CH4 /total VFA ratio with STAR, whereas CH4 /total VFA ratio tended (p < 0.10) to be decreased with CEL substrate. Increasing the amount of both proteins decreased linearly (p < 0.05) ammonia-N concentrations, which may indicate an incorporation of amino acids and peptides into microbial protein without being first deaminated into ammonia-N. In incubations with the tested N sources as the only substrate, the fermentation of 1 mg of CA or SP produced 1.24 and 0.60 μmol of CH4 respectively. The results indicate the generation of CH4 from protein fermentation, and that the response of CH4 production to protein-N supply may differ with the basal substrate.