Abstract
Nitrate supplementation into the ruminant diet can decrease ruminal methane emissions, but amounts needed to achieve appreciable decreases can risk ruminal accumulations of nitrite with potential for methemoglobinemia. A denitrifying rumen Paenibacillus fortis strain 79R4 (79R4) selected for enhanced nitrite-metabolizing ability has shown promise as a probiotic to decrease risks of nitrite toxicosis. Presently, a spray-dried prototype of this spore-forming, facultative anaerobe was tested during anaerobic culture (10 mL/tube) of rumen fluid freshly-collected from an alfalfa hay-fed cannulated Jersey cow. Cultures supplemented with 22 mM sodium nitrate and without or with inoculations of freshly-harvested (FH) cells or spray dried (SP) 79R4 spores (3 tubes/treatment; 10(8) cells/spores per tube) were cultured anaerobically (39(o)C for 24 h with 100% CO(2)). The FH- and SP- cells were grown aerobic, 72 h in tryptic soy broth, SP- cells were then processed and spray-dried. Nitrate-supplementation decreased (P = 0.0006; SEM = 0.31) methane production by the cultures, but this decrease was unaffected by 79R4 inoculation (2.57 µmol CH(4)/mL with no nitrate/no inoculum versus 0.15 µmol CH(4)/mL with nitrate/inoculum). Nitrate-metabolizing activity in nitrate-treated cultures were unaffected by 79R4 inoculations (P = 0.17; SEM = 0.15), rates being 0.95, 0.63 and 0.50 µmol nitrate/mL h(-1) FH-, SP- and non-inoculated cultures, respectively. Nitrite accumulation rates (P = 0.10; SEM = 0.06) and peak nitrite concentrations (P = 0.06; SEM = 0.75) tended to be lower in SP- than in FH- and non-inoculated cultures (0.29, 0.47 and 0.47 µmol nitrite/mL h(-1) and 3.33, 5.67 and 5.66 µmol/mL, respectively). Rates of ammonia accumulation were more rapid (P = 0.01; SEM = 0.01) in SP- and FH- than in non-inoculated cultures (0.16 and 0.15 versus 0.06 µmol/mL h(-1), respectively). Results provide evidence that 79R4 prototype may aid rumen populations in detoxifying nitrite, therefore enhancing the abilities of high nitrate diets.