Effects Of 3-nitrooxypropanol Research Articles

Effects of 3-nitrooxypropanol (Bovaer10) and whole cottonseed on milk production and enteric methane emissions from dairy cows under Swiss management conditions

The objective of this study was to determine the potential effect and interaction of 3-nitrooxypropanol (3-NOP; Bovaer, DSM-Firmenich Nutrition Products Ltd.) and whole cottonseed (WCS) on lactational performance and enteric methane (CH4) emission of dairy cows. A total of 16 multiparous cows, including 8 Holstein Friesian (HF) and 8 Brown Swiss (BS; 224 ± 36 DIM, 26 ± 3.7 kg milk yield, mean ± SD), were used in a split-plot design, where the main plot was the breed of cows. Within each subplot, cows were randomly assigned to a treatment sequence in a replicated 4 × 4 Latin square design with 2 × 2 factorial arrangements of treatments with four 24-d periods. The experimental treatments were as follows: (1) control (basal TMR), (2) 3-NOP (60 mg/kg TMR DM), (3) WCS (5% TMR DM), and (4) 3-NOP + WCS. The treatment diets were balanced for ether extract, crude protein, and NDF contents (4%, 16%, and 43% of TMR DM, respectively). The basal diets were fed twice daily at 0800 and 1800 h. Dry matter intake and milk yield were measured daily, and enteric gas emissions were measured (using the GreenFeed System, C-Lock Inc.) during the last 3 d of each 24-d experimental period when animals were housed in tiestalls. There was no difference in DMI on treatment level, whereas the WCS treatment increased ECM yield and milk fat yield. No interaction of 3-NOP and WCS occurred for any of the enteric gas emission parameters, but 3-NOP decreased CH4 production (g/d), CH4 yield (g/kg DMI), and CH4 intensity (g/kg ECM) by 13%, 14%, and 13%, respectively. Further, an unexpected interaction of breed by 3-NOP was observed for different enteric CH4 emission metrics: HF cows had a greater CH4 mitigation effect compared with BS cows for CH4 production (g/d; 18% vs. 8%), CH4 intensity (g/kg milk yield; 19% vs. 3%), and CH4 intensity (g/kg ECM; 19% vs. 4%). Hydrogen production was increased by 2.85-fold in HF and 1.53-fold in BS cows receiving 3-NOP. Further, a 3-NOP × time interaction occurred for both breeds. In BS cows, 3-NOP tended to reduce CH4 production by 18% at approximately 4 h after morning feeding, but no effect was observed at other time points. In HF cows, the greatest mitigation effect of 3-NOP (29.6%) was observed immediately after morning feeding, and it persisted at around 23% to 26% for 10 h until the second feed provision, and 3 h thereafter, in the evening. In conclusion, supplementing 3-NOP at 60 mg/kg DM to a high-fiber diet resulted in 18% to 19% reduction in enteric CH4 emission in Swiss HF cows. The lower response to 3-NOP by BS cows was unexpected and has not been observed in other studies. These results should be interpreted with caution due to the low number of cows per breed. Finally, supplementing WCS at 5% of DM improved ECM and milk fat yield but did not enhance the CH4 inhibition effect of 3-NOP of dairy cows.

The effect of 3-nitrooxypropanol, a potent methane inhibitor, on ruminal microbial gene expression profiles in dairy cows

BackgroundEnteric methane emissions from dairy cows are an environmental problem as well as a gross feed energy loss to the animal. Methane is generated in the rumen by methanogenic archaea from hydrogen (H2) + carbon dioxide and from H2 + methanol or methylamines. The methanogenic substrates are provided by non-methanogens during feed fermentation. Methane mitigation approaches have yielded variable results, partially due to an incomplete understanding of the contribution of hydrogenotrophic and methylotrophic archaea to methanogenesis. Research indicates that 3-nitrooxypropanol (3-NOP) reduces enteric methane formation in dairy cows by inhibiting methyl-coenzyme M reductase (MCR), the enzyme responsible for methane formation. The purpose of this study was to utilize metagenomic and metatranscriptomic approaches to investigate the effect of 3-NOP on the rumen microbiome and to determine the fate of H2 that accumulates less than expected under inhibited methanogenesis.ResultsThe inhibitor 3-NOP was more inhibitory on Methanobrevibacter species than methanol-utilizing Methanosphaera and tended to reduce the gene expression of MCR. Under inhibited methanogenesis by 3-NOP, fluctuations in H2 concentrations were accompanied by changes in the expression of [FeFe] hydrogenases in H2-producing bacteria to regulate the amount of H2 production. No previously reported alternative H2 sinks increased under inhibited methanogenesis except for a significant increase in gene expression of enzymes involved in the butyrate pathway.ConclusionBy taking a metatranscriptomic approach, this study provides novel insights on the contribution of methylotrophic methanogens to total methanogenesis and regulation of H2 metabolism under normal and inhibited methanogenesis by 3-NOP in the rumen.-iX6YPbp4qKjzQQ25nvbCQVideo

Effects of 3-nitrooxypropanol and varying concentrate feed proportions in the ration on methane emission, rumen fermentation and performance of periparturient dairy cows

ABSTRACT The climate-relevant enteric methane (CH4) formation represents a loss of feed energy that is potentially meaningful for energetically undersupplied peripartal dairy cows. Higher concentrate feed proportions (CFP) are known to reduce CH4 emissions in cows. The same applies to the feed additive 3-nitrooxypropanol (3-NOP), albeit through different mechanisms. It was hypothesised that the hydrogen not utilised for CH4 formation through the inhibition by 3-NOP would be sequestered by propionate formation triggered by higher CFP so that it could thereby give rise to a synergistically reduced CH4 emission. In a 2 × 2-factorial design, low (LC) or high (HC) CFP were either tested without supplements (CONLC, CONHC) or combined with 3-NOP (NOPLC, 48.4 mg/kg dry matter (DM); NOPHC, 51.2 mg 3-NOP/kg DM). These four rations were fed to a total of 55 Holstein cows from d 28 ante partum until d 120 post partum. DM intake (DMI) was not affected by 3-NOP but increased with CFP (CFP; p < 0.001). CH4/DMI and CH4/energy-corrected milk (ECM) were mitigated by 3-NOP (23% NOPLC, 33% NOPHC) (p < 0.001) and high CFP (12% CON, 22% 3-NOP groups) (CFP × TIME p < 0.001). Under the conditions of the present experiment, the CH4 emissions of NOPLC increased to the level of the CON groups from week 8 until the end of trial (3-NOP × CFP × TIME; p < 0.01). CO2 yield decreased by 3-NOP and high CFP (3-NOP × CFP; p < 0.001). The reduced body weight loss and feed efficiency in HC groups paralleled a more positive energy balance being most obvious in NOPHC (3-NOP × CFP; p < 0.001). ECM was lower for NOPHC compared to CONHC (3-NOP × CFP; p < 0.05), whereas LC groups did not differ. A decreased fat to protein ratio was observed in HC groups and, until week 6 post partum, in NOPLC. Milk lactose and urea increased by 3-NOP (3-NOP; p < 0.05). 3-NOP and high CFP changed rumen fermentation to a more propionic-metabolic profile (3-NOP; CFP; p < 0.01) but did not affect rumen pH. In conclusion, CH4 emission was synergistically reduced when high CFP was combined with 3-NOP while the CH4 mitigating 3-NOP effect decreased with progressing time when the supplement was added to the high-forage ration. The nature of these interactions needs to be clarified.

Effects of 3-nitrooxypropanol on enteric methane production, rumen fermentation, and feeding behavior in beef cattle fed a high-forage or high-grain diet1.

The objective of the study was to determine whether feeding a diet supplemented with 3-nitrooxypropanol (3-NOP) affects feeding behavior altering intake and rumen fermentation. Two experiments were conducted with 9 rumen-cannulated beef steers in a replicated 3 × 3 Latin square design where animals received a high-forage or high-grain diet. Treatments were 1) a basal diet (CON), the CON diet supplemented with 3-NOP (dNOP; 100 mg/kg in dietary DM or 1 g/d), or the CON diet with 3-NOP (1 g/d) infused into the rumen (infNOP). Each experimental period consisted of 14-d diet adaptation and 7-d sample collection. A 7-d washout period was provided between experiment periods. All data were analyzed as a Latin square design using Mixed Procedure of SAS. In Exp. 1 (high-forage diet), methane yield (measured by the Greenfeed system) was lowered by 18% (18.6 vs. 22.7 g/kg DMI; P < 0.01) by dNOP compared with CON. Rumen fermentation was altered similarly by both NOP treatments compared with CON where dNOP and infNOP increased (P < 0.01) rumen pH at 3 h and decreased (P < 0.01) proportion of acetate in total VFA. However, DMI, feed consumption rate (0 to 3, 3 to 6, 6 to 12, and 12 to 24 h after feeding), particle size distribution of orts, and feeding behavior (videotaped for individual animals over 48 h) were not affected by dNOP and infNOP compared with CON. In Exp. 2 (high-grain diet), methane production was not affected by dNOP or infNOP compared with CON. Dry matter intake, feed consumption rate, particle size distribution of orts, and feeding behavior were not altered by dNOP and infNOP compared with CON. However, both dNOP and infNOP affected rumen fermentation where total VFA decreased (P = 0.04) and acetate proportion in total VFA tended to decrease (P = 0.07) compared with CON. In conclusion, dietary supplementation of 3-NOP did not affect feeding behavior of beef steers fed a high-forage or high-grain diet. However, rumen fermentation was similarly changed when 3-NOP was provided in the diet or directly infused in the rumen. Thus, observed changes in rumen fermentation with 3-NOP were not due to changes in feeding behavior indicating no effects on the organoleptic property of the diets. In addition, according to small or no changes in DMI in both experiments and relatively small changes in rumen fermentation in Exp. 2, a greater dosage level of 3-NOP than 100 mg/kg (dietary DM) may need further examination of its effects on feeding behavior of beef cattle.

Use of 3-nitrooxypropanol as feed additive for mitigating enteric methane emissions from ruminants: a meta-analysis

This study aimed to perform a meta-analysis on the effect of 3-nitrooxypropanol (3-NOP) on enteric methane (CH4) emissions from ruminants. A total of 12 in vivo studies from 10 articles were integrated into a database. Ruminant species included were dairy cows, beef cattle and sheep. Concentration of 3-NOP in diets varied from 0 to 280 mg/kg dry matter intake (DMI). Parameters included were CH4 emissions, rumen fermentation, microbial population, nutrient digestibility and animal performance. Meta-analysis of data was performed by using mixed model methodology in which different studies were treated as random effects whereas 3-NOP addition levels in diets of ruminants were treated as fixed effects. Results showed that increasing level of 3-NOP addition in diets of ruminants decreased enteric CH4 emissions per unit of body weight, CH4/DMI, CH4/milk produced, CH4/digested organic matter or CH4/gross energy intake (p < .05). Production of H2 was higher with increasing level of 3-NOP addition (p < .001). Addition of 3-NOP decreased total VFA concentration (p < .01), and decreased and increased proportions of C2 and C3, respectively (p < .001). Addition of 3-NOP decreased archaea population (p < .01) but it did not change total bacteria and protozoa populations. The substance had minor effect on digestibility of nutrients. Production performance of dairy cows and beef cattle was limitedly influenced by the addition of 3-NOP in the diets, and it had no negative effect on DMI of ruminants. It is concluded that 3-NOP is an effective feed additive to mitigate enteric CH4 emissions without compromising productive performance of ruminants.

Rapid Communication: Evaluation of methane inhibitor 3-nitrooxypropanol and monensin in a high-grain diet using the rumen simulation technique (Rusitec)

The objective of this study was to evaluate the effects of 3-nitrooxypropanol (NOP), a known methane (CH) inhibitor; the ionophore monensin (MON); and their combination on in vitro CH production in a high-grain diet (85% barley grain, 10% barley silage, and 5% vitamin-mineral supplement; DM basis) using a rumen simulation technique (Rusitec). Sixteen fermentation vessels in 2 Rusitec apparatuses (blocks) were used in a completely randomized block design with 4 treatments: Control, NOP (200 µg/g DM), MON (200 µg/g DM), and the combination of 200 µg NOP/g DM and 200 µg MON/g DM (NOP + MON). Two fermenters within each apparatus were randomly assigned to a treatment. Treatments were mixed with 10 g of substrate and supplied on a daily basis. The study included an 8-d adaptation period without treatment supplementation and a 6-d period for addition of treatments. Dry matter disappearance, pH, and total VFA were not affected by treatment ( ≥ 0.34). Acetate proportion was decreased by 8.3% and 14.9% with NOP and NOP + MON ( < 0.01), respectively; however, propionate proportion was not affected by treatment ( = 0.44). The acetate to propionate ratio was lowered by 21.1% with the combination of NOP and MON ( = 0.02), whereas ammonia-N concentration was not affected by treatment ( = 0.50). Total gas production was unaffected ( = 0.50), but CH production decreased by 77.7% and 75.95% ( < 0.01) with NOP and NOP + MON addition, respectively. Concurrently, H gas production increased by 131.3% and 185.6% ( = 0.01) with NOP and NOP + MON treatments, respectively. The copy number of methanogens was decreased in both solid and liquid phases ( < 0.01) with NOP and NOP + MON treatments. Despite the combination of NOP + MON showing the greatest decrease in acetate molar proportion and acetate to propionate ratio, it did not further inhibit CH beyond the effect of NOP alone. The decrease in CH emissions with treatments that included NOP occurred along with a decrease in the copy number of methanogens associated with the solid and liquid phases, confirming the inhibitory effects of NOP on these microorganisms. In conclusion, the combined effects of NOP and MON on CH mitigation did not exceed the effect of NOP alone when using a high-grain diet in vitro.

Effects of 3-nitrooxypropanol and monensin on methane production using a forage-based diet in Rusitec fermenters

3-Nitrooxypropanol (NOP) is an enzymatic inhibitor that has consistently decreased enteric methane (CH4) emissions in ruminants while monensin (MON) has a moderate, and sometimes transitory, effect on CH4 decrease. We hypothesised that combining NOP and MON would result in an additive CH4 decrease. The objective was to evaluate the effect of NOP (2mg), MON (2mg) and the combination of NOP (2mg) and MON (2mg) on CH4 production using a forage-based diet (600g/kg barley silage, 350g/kg barley grain and 50g/kg vitamin-mineral supplement; dry matter basis) incubated in Rusitec fermenters. The experiment included an adaptation period without treatment supplementation (8 d), a treatment period (7 d), and a recovery period where treatments were discontinued (3 d). During the treatment period CH4 production (mL/d) was decreased (P<0.01) by 71.5, 11.8 and 69.9% with NOP, MON and NOP+MON respectively. Hydrogen gas (H2) production was increased (P<0.01) by 66.4 and 82.5% with NOP and NOP+MON respectively. The total copy number of the 16S rRNA gene for methanogens (log10 copies/mL) in the solid fraction, obtained using quantitative PCR, was decreased (P<0.01) by 17.0 and 21.4% for NOP and NOP+MON, respectively. During the recovery period, CH4 production remained lower than control (P<0.01) with NOP and NOP+MON treatments; however, a gradual increase in CH4 concentration was observed after treatment withdrawal. No effect (P=0.53) on H2 production was observed during the recovery period. In conclusion, addition of NOP and, to a lesser extent, MON, to a forage-based diet is an effective means of decreasing CH4 production in vitro; however, the combination of both did not produce an additive effect.

Effect of 3-nitrooxypropanol on methane and hydrogen emissions, methane isotopic signature, and ruminal fermentation in dairy cows

The objective of this crossover experiment was to investigate the effect of a methane inhibitor, 3-nitrooxypropanol (3NOP), on enteric methane emission, methane isotopic composition, and rumen fermentation and microbial profile in lactating dairy cows. The experiment involved 6 ruminally cannulated late-lactation Holstein cows assigned to 2 treatments: control and 3NOP (60 mg/kg of feed dry matter). Compared with the control, 3NOP decreased methane emission by 31% and increased hydrogen emission from undetectable to 1.33 g/d. Methane emissions per kilogram of dry matter intake and milk yield were also decreased 34% by 3NOP. Milk production and composition were not affected by 3NOP, except milk fat concentration was increased compared with the control. Concentrations of total VFA and propionate in ruminal fluid were not affected by treatment, but acetate concentration tended to be lower and acetate-to-propionate ratio was lower for 3NOP compared with the control. The 3NOP decreased the molar proportion of acetate and increase those of propionate, butyrate, valerate, and isovalerate. Deuterium-to-hydrogen ratios of methane and the abundance of 13CH3D were similar between treatments. Compared with the control, minor (4‰) depletion in the 13C/12C ratio was observed for 3NOP. Genus composition of methanogenic archaea (Methanobrevibacter, Methanosphaera, and Methanomicrobium) was not affected by 3NOP, but the proportion of methanogens in the total cell counts tended to be decreased by 3NOP. Prevotella spp., the predominant bacterial genus in ruminal contents in this experiment, was also not affected by 3NOP. Compared with the control, Ruminococcus and Clostridium spp. were decreased and Butyrivibrio spp. was increased by 3NOP. This experiment demonstrated that a substantial inhibition of enteric methane emission by 3NOP in dairy cows was accompanied with increased hydrogen emission and decreased acetate-to-propionate ratio; however, neither an effect on rumen archaeal community composition nor a significant change in the isotope composition of methane was observed.