J. Takahashi

Effect of ruminal administration of Escherichia coli wild type or a genetically modified strain with enhanced high nitrite reductase activity on methane emission and nitrate toxicity in nitrate-infused sheep.

The effects of two kinds of Escherichia coli (E. coli) strain, wild-type E. coli W3110 and E. coli nir-Ptac, which has enhanced NO(2) reduction activity, on oral CH(4) emission and NO(3) toxicity in NO(3)-treated sheep were assessed in a respiratory hood system in a 4 x 6 Youden square design. NO(3) (1.3 g NaNO(3)/kg(0.75) body weight) and/or E. coli strains were delivered into the rumen through a fistula as a single dose 30 min after the morning meal. Escherichia coli cells were inoculated for sheep to provide an initial E. coli cell density of optical density at 660 nm of 2, which corresponded to 2 x 10(10) cells/ml. The six treatments consisted of saline, E. coli W3110, E. coli nir-Ptac, NO(3), NO(3) plus E. coli W3110, and NO(3) plus E. coli nir-Ptac. CH(4) emission from sheep was reduced by the inoculation of E. coli W3110 or E. coli nir-Ptac by 6 % and 12 %, respectively. NO(3) markedly inhibited CH(4) emission from sheep. Compared with sheep given NO(3) alone, the inoculation of E. coli W3110 to NO(3)-infused sheep lessened ruminal and plasma toxic NO(2) accumulation and blood methaemoglobin production, while keeping ruminal methanogenesis low. Ruminal and plasma toxic NO(2) accumulation and blood methaemoglobin production in sheep were unaffected by the inoculation of E. coli nir-Ptac. These results suggest that ruminal methanogenesis may be reduced by the inoculation of E. coli W3110 or E. coli nir-Ptac. The inoculation of E. coli W3110 may abate NO(3) toxicity when NO(3) is used to inhibit CH(4) emission from ruminants.

Effect of nisin on ruminal methane production and nitrate/nitrite reduction in vitro

The suppressing effects of different concentrations of nitrate (0, 5, 10, 15, and 20 mm) or nisin (0, 5, 10, 15, 20, and 30 μmol/L) on in vitro methane production were examined with mixed rumen microbes using the in vitro continuous incubation system. The effects of different concentrations of nisin (10, 20, and 30 μmol/L) on in vitro nitrate/nitrite reduction were examined for methane suppression without any nitrate toxicity. The culture mixture consisted of 400 mL of strained rumen fluid from 2 non-lactating Holstein cows fed a diet of oaten hay, alfalfa hay cube, and concentrates (35 : 35 : 30) at maintenance level, and 400 mL of autoclaved buffer solution. Methane production was decreased with increasing levels of nitrate. As the concentration of nisin increased from 5 to 30 μmol/L, methane production was decreased by 14–40%. A decrease in acetate to propionate ratio and increase in total volatile fatty acids were observed as the concentration of nisin increased. Toxic nitrite accumulation was unaffected by increasing levels of nisin. In conclusion, nisin improved some of the parameters of ruminal fermentation and inhibited methane production, but did not decrease nitrate toxicity when nitrate was used to inhibit methane production.