The effect of incremental levels of dietary nitrate on methane emissions in Holstein steers and performance in Nelore bulls.

Two experiments were conducted to study effects of dietary nitrate on enteric methane production, blood methemoglobin concentration, and growth rate in cattle. In Exp. 1, 36 Holstein steers (288 ± 25 kg BW) were fed increasing levels of dietary nitrate (6 levels; 0 to 3.0% of feed DM) in corn silage-based total mixed rations. Nitrate was introduced gradually in a 25-d adaptation period before methane production was determined in environmentally controlled rooms. In the rooms, feed intake was restricted and similar among all treatments. Methane production (g/d) decreased linearly as dietary nitrate concentration increased (P < 0.01). The apparent efficiency (measured methane reduction divided by potential methane reduction) with which enteric methane was mitigated was 49%. Blood methemoglobin levels increased with increasing nitrate dose. In Exp. 2, 300 Nelore bulls (392 ± 28 kg) were fed increasing levels of nitrate (6 levels; 0 to 2.4% of feed DM) in high-concentrate total mixed rations offered ad libitum. Feed intake decreased linearly with increasing level of dietary nitrate (P < 0.01). However, ADG was not affected by nitrate dose (P = 0.54), resulting in a linear improvement in G:F (P = 0.03) as dietary nitrate level increased. Carcass dressing percentage showed a quadratic response to incremental dietary nitrate, reaching the highest value at 0.96% of NO3/kg DM (P = 0.04).

Evaluation of the SF6 tracer technique for estimating methane emission rates with reference to dairy cows using a mechanistic model.

A dynamic, mechanistic model of the sulfur hexafluoride (SF6) tracer technique, used for estimating methane (CH4) emission rates from ruminants, was constructed to evaluate the accuracy of the technique. The model consists of six state variables and six zero-pools representing the quantities of SF6 and CH4 in rumen and hindgut fluid, in rumen and hindgut headspace, and in blood and collection canister. The model simulates flows of CH4 and SF6 through the body, subsequent eructation and exhalation and accumulation in a collection canister. The model predicts CH4 emission by multiplying the SF6 release rate of a permeation device in the rumen by the ratio of CH4:SF6 in collected air. This prediction is compared with the actual CH4 production rate, assumed to be continuous and used as a driving variable in the model. A sensitivity analysis was conducted to evaluate the effect of changes in several parameters. The predicted CH4 emission appeared sensitive to parameters affected by the difference in CH4:SF6 ratio in exhaled and eructed air respectively, viz., hindgut fractional passage rate and hindgut CH4 production. This is caused by the difference in solubility of CH4 and SF6 and by hindgut CH4 production. In addition, the predicted CH4 emission rate appeared sensitive to factors that affect proportions of exhaled and eructed air sampled, i.e., eructation time fraction, exhalation time fraction, and distance from sampling point to mouth/nostrils. Changes in rumen fractional passage rate, eructation rate, SF6 release rate, background values and air sampling rate did not noticeably affect the predicted CH4 emission. Simulations with (13)CH4 as an alternative tracer show that the differences and sensitivity to parameters greatly disappear. The model is considered a useful tool to evaluate critical points in the SF6 technique. Data from in vivo experiments are needed to further evaluate model simulations.

Dietary nitrate supplementation reduces methane emission in beef cattle fed sugarcane-based diets.

The objective of this study was to determine the effect of dietary nitrate on methane emission and rumen fermentation parameters in Nellore × Guzera (Bos indicus) beef cattle fed a sugarcane based diet. The experiment was conducted with 16 steers weighing 283 ± 49 kg (mean ± SD), 6 rumen cannulated and 10 intact steers, in a cross-over design. The animals were blocked according to BW and presence or absence of rumen cannula and randomly allocated to either the nitrate diet (22 g nitrate/kg DM) or the control diet made isonitrogenous by the addition of urea. The diets consisted of freshly chopped sugarcane and concentrate (60:40 on DM basis), fed as a mixed ration. A 16-d adaptation period was used to allow the rumen microbes to adapt to dietary nitrate. Methane emission was measured using the sulfur hexafluoride tracer technique. Dry matter intake (P = 0.09) tended to be less when nitrate was present in the diet compared with the control, 6.60 and 7.05 kg/d DMI, respectively. The daily methane production was reduced (P < 0.01) by 32% when steers were fed the nitrate diet (85 g/d) compared with the urea diet (125 g/d). Methane emission per kilogram DMI was 27% less (P < 0.01) on the nitrate diet (13.3 g methane/kg DMI) than on the control diet (18.2 g methane/kg DMI). Methane losses as a fraction of gross energy intake (GEI) were less (P < 0.01) on the nitrate diet (4.2% of GEI) than on the control diet (5.9% of GEI). Nitrate mitigated enteric methane production by 87% of the theoretical potential. The rumen fluid ammonia-nitrogen (NH(3)-N()) concentration was significantly greater (P < 0.05) for the nitrate diet. The total concentration of VFA was not affected (P = 0.61) by nitrate in the diet, while the proportion of acetic acid tended to be greater (P = 0.09), propionic acid less (P = 0.06) and acetate/propionate ratio tended to be greater (P = 0.06) for the nitrate diet. Dietary nitrate reduced enteric methane emission in beef cattle fed sugarcane based diet.

Persistency of methane mitigation by dietary nitrate supplementation in dairy cows.

Feeding nitrate to dairy cows may lower ruminal methane production by competing for reducing equivalents with methanogenesis. Twenty lactating Holstein-Friesian dairy cows (33.2±6.0 kg of milk/d; 104±58 d in milk at the start of the experiment) were fed a total mixed ration (corn silage-based; forage to concentrate ratio 66:34), containing either a dietary urea or a dietary nitrate source [21 g of nitrate/kg of dry matter (DM)] during 4 successive 24-d periods, to assess the methane-mitigating potential of dietary nitrate and its persistency. The study was conducted as paired comparisons in a randomized design with repeated measurements. Cows were blocked by parity, lactation stage, and milk production at the start of the experiment. A 4-wk adaptation period allowed the rumen microbes to adapt to dietary urea and nitrate. Diets were isoenergetic and isonitrogenous. Methane production, energy balance, and diet digestibility were measured in open-circuit indirect calorimetry chambers. Cows were limit-fed during measurements. Nitrate persistently decreased methane production by 16%, whether expressed in grams per day, grams per kilogram of dry matter intake (DMI), or as percentage of gross energy intake, which was sustained for the full experimental period (mean 368 vs. 310±12.5 g/d; 19.4 vs. 16.2±0.47 g/kg of DMI; 5.9 vs.4.9±0.15% of gross energy intake for urea vs. nitrate, respectively). This decrease was smaller than the stoichiometrical methane mitigation potential of nitrate (full potential=28% methane reduction). The decreased energy loss from methane resulted in an improved conversion of dietary energy intake into metabolizable energy (57.3 vs. 58.6±0.70%, urea vs. nitrate, respectively). Despite this, milk energy output or energy retention was not affected by dietary nitrate. Nitrate did not affect milk yield or apparent digestibility of crude fat, neutral detergent fiber, and starch. Milk protein content (3.21 vs. 3.05±0.058%, urea vs. nitrate respectively) but not protein yield was lower for dietary nitrate. Hydrogen production between morning and afternoon milking was measured during the last experimental period. Cows fed nitrate emitted more hydrogen. Cows fed nitrate displayed higher blood methemoglobin levels (0.5 vs. 4.0±1.07% of hemoglobin, urea vs. nitrate respectively) and lower hemoglobin levels (7.1 vs. 6.3±0.11 mmol/L, urea vs. nitrate respectively). Dietary nitrate persistently decreased methane production from lactating dairy cows fed restricted amounts of feed, but the reduction in energy losses did not improve milk production or energy balance.

Dietary inclusion of diallyl disulfide, yucca powder, calcium fumarate, an extruded linseed product, or medium-chain fatty acids does not affect methane production in lactating dairy cows.

Two similar experiments were conducted to assess the effect of diallyl disulfide (DADS), yucca powder (YP), calcium fumarate (CAFU), an extruded linseed product (UNSAT), or a mixture of capric and caprylic acid (MCFA) on methane production, energy balance, and dairy cow performance. In experiment 1, a control diet (CON1) and diets supplemented with 56 mg of DADS/kg of dry matter (DM), 3g of YP/kg of DM, or 25 g of CAFU/kg of DM were evaluated. In experiment 2, an inert saturated fat source in the control diet (CON2) was exchanged isolipidically for an extruded linseed source (100g/kg of DM; UNSAT) or a mixture of C8:0 and C10:0 (MCFA; 20.3g/kg of DM). In experiment 2, a higher inclusion level of DADS (200mg/kg of DM) was also tested. Both experiments were conducted using 40 lactating Holstein-Friesian dairy cows. Cows were adapted to the diet for 12 d and were subsequently kept in respiration chambers for 5 d to evaluate methane production, diet digestibility, energy balance, and animal performance. Feed intake was restricted to avoid confounding effects of possible differences in ad libitum feed intake on methane production. Feed intake was, on average, 17.5 and 16.6 kg of DM/d in experiments 1 and 2, respectively. None of the additives reduced methane production in vivo. Methane production in experiment 1 was 450, 453, 446, and 423 g/d for CON1 and the diets supplemented with DADS, YP, and CAFU, respectively. In experiment 2, methane production was 371, 394, 388, and 386 g/d for CON2 and the diets supplemented with UNSAT, MCFA, and DADS, respectively. No effects of the additives on energy balance or neutral detergent fiber digestibility were observed. The addition of MCFA increased milk fat content (5.38% vs. 4.82% for control) and fat digestibility (78.5% vs. 59.8% for control), but did not affect milk yield or other milk components. The other products did not affect milk yield or composition. Results from these experiments emphasize the need to confirm methane reductions observed in vitro with in vivo data.

Effects of a combination of feed additives on methane production, diet digestibility, and animal performance in lactating dairy cows.

Two experiments were conducted to assess the effects of a mixture of dietary additives on enteric methane production, rumen fermentation, diet digestibility, energy balance, and animal performance in lactating dairy cows. Identical diets were fed in both experiments. The mixture of feed additives investigated contained lauric acid, myristic acid, linseed oil, and calcium fumarate. These additives were included at 0.4, 1.2, 1.5, and 0.7% of dietary dry matter, respectively (treatment ADD). Experimental fat sources were exchanged for a rumen inert source of fat in the control diet (treatment CON) to maintain isolipidic rations. Cows (experiment 1, n=20; experiment 2, n=12) were fed restricted amounts of feed to avoid confounding effects of dry matter intake on methane production. In experiment 1, methane production and energy balance were studied using open-circuit indirect calorimetry. In experiment 2, 10 rumen-fistulated animals were used to measure rumen fermentation characteristics. In both experiments animal performance was monitored. The inclusion of dietary additives decreased methane emissions (g/d) by 10%. Milk yield and milk fat content tended to be lower for ADD in experiment 1. In experiment 2, milk production was not affected by ADD, but milk fat content was lower. Fat- and protein-corrected milk was lower for ADD in both experiments. Milk urea nitrogen content was lowered by ADD in experiment 1 and tended to be lower in experiment 2. Apparent total tract digestibility of fat, but not that of starch or neutral detergent fiber, was higher for ADD. Energy retention did not differ between treatments. The decrease in methane production (g/d) was not evident when methane emission was expressed per kilogram of milk produced. Feeding ADD resulted in increases of C12:0 and C14:0 and the intermediates of linseed oil biohydrogenation in milk in both experiments. In experiment 2, ADD-fed cows tended to have a decreased number of protozoa in rumen fluid when compared with that in control cows. Total volatile fatty acid concentrations were lower for ADD, whereas molar proportions of propionate increased at the expense of acetate and butyrate.

Nitrate and sulfate: Effective alternative hydrogen sinks for mitigation of ruminal methane production in sheep.

Twenty male crossbred Texel lambs were used in a 2 × 2 factorial design experiment to assess the effect of dietary addition of nitrate (2.6% of dry matter) and sulfate (2.6% of dry matter) on enteric methane emissions, rumen volatile fatty acid concentrations, rumen microbial composition, and the occurrence of methemoglobinemia. Lambs were gradually introduced to nitrate and sulfate in a corn silage-based diet over a period of 4 wk, and methane production was subsequently determined in respiration chambers. Diets were given at 95% of the lowest ad libitum intake observed within one block in the week before methane yield was measured to ensure equal feed intake of animals between treatments. All diets were formulated to be isonitrogenous. Methane production decreased with both supplements (nitrate: -32%, sulfate: -16%, and nitrate+sulfate: -47% relative to control). The decrease in methane production due to nitrate feeding was most pronounced in the period immediately after feeding, whereas the decrease in methane yield due to sulfate feeding was observed during the entire day. Methane-suppressing effects of nitrate and sulfate were independent and additive. The highest methemoglobin value observed in the blood of the nitrate-fed animals was 7% of hemoglobin. When nitrate was fed in combination with sulfate, methemoglobin remained below the detection limit of 2% of hemoglobin. Dietary nitrate decreased heat production (-7%), whereas supplementation with sulfate increased heat production (+3%). Feeding nitrate or sulfate had no effects on volatile fatty acid concentrations in rumen fluid samples taken 24h after feeding, except for the molar proportion of branched-chain volatile fatty acids, which was higher when sulfate was fed and lower when nitrate was fed, but not different when both products were included in the diet. The total number of rumen bacteria increased as a result of sulfate inclusion in the diet. The number of methanogens was reduced when nitrate was fed. Enhanced levels of sulfate in the diet increased the number of sulfate-reducing bacteria. The number of protozoa was not affected by nitrate or sulfate addition. Supplementation of a diet with nitrate and sulfate is an effective means for mitigating enteric methane emissions from sheep.