Dietary nitrate and presence of protozoa increase nitrate and nitrite reduction in the rumen of sheep.

Nitrate ( ) supplementation is an effective methane (CH ) mitigation strategy for ruminants but may produce nitrite ( ) toxicity. It has been reported that rumen protozoa have greater ability for and reduction than bacteria. It was hypothesised that the absence of ruminal protozoa in sheep may lead to higher accumulation in the rumen and a higher blood methaemoglobin (MetHb) concentration. An in vivo experiment was conducted with defaunated (DEF) and faunated (FAU) sheep supplemented with 1.8% in DM. The effects of rumen protozoa on concentrations of plasma and ruminal and , blood MetHb, ruminal volatile fatty acid (VFA) and ruminal ammonia (NH ) were investigated. Subsequently, two in vitro experiments were conducted to determine the contribution of protozoa to and reduction rates in DEF and FAU whole rumen digesta (WRD) and its liquid (LIQ) and solid (SOL) fractions, incubated alone (CON), with the addition of or with the addition of . The results from the in vivo experiment showed no differences in total VFA concentrations, although ruminal NH was greater (p < .01) in FAU sheep. Ruminal , and plasma concentrations tended to increase (p < .10) 1.5 hr after feeding in FAU relative to DEF sheep. In vitro results showed that reduction to NH was stimulated (p < .01) by incoming in both DEF and FAU relative to CON digesta. However, adding increased (p < .05) the rate of accumulation in the SOL fraction of DEF relative to both fractions of FAU digesta. Results observed in vivo and in vitro suggest that and are more rapidly metabolised in the presence of rumen protozoa. Defaunated sheep may have an increased risk of poisoning due to accumulation in the rumen.