An inhibitor persistently decreased enteric methane emission from dairy cows with no negative effect on milk production

A quarter of all anthropogenic methane emissions in the United States are from enteric fermentation, primarily from ruminant livestock. This study was undertaken to test the effect of a methane inhibitor, 3-nitrooxypropanol (3NOP), on enteric methane emission in lactating Holstein cows. An experiment was conducted using 48 cows in a randomized block design with a 2-wk covariate period and a 12-wk data collection period. Feed intake, milk production, and fiber digestibility were not affected by the inhibitor. Milk protein and lactose yields were increased by 3NOP. Rumen methane emission was linearly decreased by 3NOP, averaging about 30% lower than the control. Methane emission per unit of feed dry matter intake or per unit of energy-corrected milk were also about 30% less for the 3NOP-treated cows. On average, the body weight gain of 3NOP-treated cows was 80% greater than control cows during the 12-wk experiment. The experiment demonstrated that the methane inhibitor 3NOP, applied at 40 to 80 mg/kg feed dry matter, decreased methane emissions from high-producing dairy cows by 30% and increased body weight gain without negatively affecting feed intake or milk production and composition. The inhibitory effect persisted over 12 wk of treatment, thus offering an effective methane mitigation practice for the livestock industries.The livestock sector is a significant source of greenhouse gas (GHG) emissions in the United States and globally (1, 2). In the United States, enteric fermentation of feed by ruminants is the largest source of anthropogenic methane emissions (0.14 Gt of CO₂ Eq. in 2012; or 25% of the total methane emissions; ref. 3). Globally, according to the most recent Intergovernmental Panel on Climate Change (IPCC) report, GHG emissions from agriculture represent around 10–12% (5.0–5.8 Gt CO₂ Eq/yr) of the total anthropogenic GHG emissions (1). In this report, livestock contribution to the global anthropogenic GHG emissions was estimated at 6.3%, with GHG emissions from enteric fermentation accounting for 2.1 Gt CO₂ Eq/yr and manure management accounting for 0.99 Gt CO₂ Eq/yr (1). The relative contribution of emissions from enteric fermentation to the total agricultural GHG emissions will vary by region depending on the structure of agricultural production and type of livestock production systems. For example, GHG from enteric fermentation were estimated at 57% for New Zealand, a country with a large, pasture-based livestock sector (4). Extensive research in recent years has provided a number of viable enteric methane mitigation practices, such as alternative electron receptors, methane inhibitors, dietary lipids, and increased animal productive efficiency (5). Methane emission in the reticulo-rumen is an evolutionary adaptation that enables the rumen ecosystem to dispose of hydrogen, a fermentation product and an important energy substrate for the methanogenic archaea (6), which may otherwise accumulate and inhibit carbohydrate fermentation and fiber degradation (7, 8). Some compounds may be effective in decreasing methane emission, but they may also decrease feed intake, fiber degradability, and animal productivity (5), or the rumen archaea may adapt to them (9). Therefore, it is important to evaluate methane mitigation strategies in long-term experiments, which for livestock experimentation requires treatment periods considerably longer than the 21–28 d, common for crossover designs. In addition, due to a variety of constraints and confounding factors of batch or continuous culture in vitro systems (5, 10), mitigation compounds, including methane inhibitors, have to be tested in vivo using animals with similar productivity to those on commercial farms. An example of the limitations of in vitro systems is a series of experiments with garlic oil. In continuous rumen culture, garlic oil was very effective in inhibiting rumen methane emission (11), but it failed to produce an effect in sheep (12). The nutrient requirements of high-producing dairy cows are much greater than those of nonlactating or low-producing cows (13) and hence any reduction in feed intake caused by a methane mitigation compound or practice would likely result in decreased productivity, which may not be evident in low-producing cows.Methane inhibitors are chemical compounds with inhibitory effects on rumen archaea. Compounds such as bromochloromethane, 2-bromoethane sulfonate, chloroform, and cyclodextrin have been tested, some successfully, in various ruminant species (5). Inhibition of methanogenesis by these compounds in vivo can be up to 60% with the effect of bromochloromethane shown to persist in long-term experiments (5, 14). However, the viability of these compounds as mitigation agents has been questioned due to concerns for animal health, food safety, or environmental impact. Bromochloromethane, for example, is an ozone-depleting agent and is banned in many countries.Among the efficacious methane inhibitors identified is 3-nitrooxypropanol (3NOP; ref. 15). This compound was part of a developmental program designing specific small molecule inhibitors for methyl coenzyme-M (CoM) reductase, the enzyme that catalyzes the last step of methanogenesis, the reduction of methyl CoM and coenzyme-B (CoB) into methane and a CoM–CoB complex (16). A continuous in vitro culture study (11) was followed by in vivo experiments in sheep (17), beef (18), and dairy cattle (19, 20), which demonstrated that 3NOP is an effective methane inhibitor. However, these experiments were conducted using nonlactating animals (17), or were short-term (<35 d; refs. 19 and 20). The rumen microorganisms have the ability to adapt to foreign agents or changes in the feeding regimen and, therefore, short-term responses are not representative of the effect of a given mitigation compound or practice in real farm conditions. McIntosh et al. (21), for example, showed that the MIC50 of essential oils doubled or tripled for a number of important rumen bacteria (Butyrivibrio fibrisolvens, Prevotella bryantii, Ruminococcus albus, Ruminobacter amylophilus), if they were adapted to the treatment for a period of 10 d. Thus, it is critically important for the success of GHG mitigation efforts to substantiate the mitigation potential of a given compound in long-term animal experiments before considering it for adoption by the livestock industries.