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.     

图形翻转视觉诱发电位在视神经管减压后视神经损伤不同时期的变化特征

目的:观察视神经损伤动物模型在损伤后和不同时期视神经管减压后视觉诱发电位的变化,了解创伤性视神经损伤的手术时机与疗效的关系。方法:实验于2005-03/05在解放军南京军区南京总医院动物实验中心完成。①实验分组:30只新西兰白兔随机分为正常对照组、术后2d减压组、术后7d减压组、术后14d减压组、术后不减压组,每组6只。②造模:除正常对照组外,其余各组在视神经孔中塞入一细端为2mm直径的圆锥软硅胶,阻塞视神经孔,造成视神经的挤压伤。③指标检测:采用图形翻转视觉诱发电位检测损伤前、损伤后1h、减压前1h、减压后2周视功能变化,记录NPN曲线主波(P波)的绝对潜伏期、绝对波幅。正常对照组仅采集一组数据作为对照。结果:30只实验动物均进入结果分析。①正常对照组家兔图形翻转视觉诱发电位检查均引出典型NPN波型曲线,视神经挤压伤后1hNPN波形低阔扁平,P波潜伏期延长,波幅降低。②P波潜伏期:术后2d减压组减压后短于减压前[(71.25±8.51),(86.47±14.28)ms,P<0.05];术后7d减压组减压前后比较差异无显著性(P>0.05);术后14d减压组减压后明显长于减压前[(158.73±15.16),(116.35±17.13)ms,P<0.05]。术后2d减压组和术后7d减压组短于术后不减压组(P<0.01)。术后7,14d减压组和术后不减压组明显长于正常对照组(P<0.01)。③P波波幅:术后2d减压组减压后高于减压前[(5.25±0.78),(4.42±0.42)μV,P<0.05]。术后2d减压组减压后低于术后7d减压组、术后14d减压组(P<0.01),术后14d减压组低于术后7d减压组(P<0.05);术后7d减压组、术后14d减压组、术后不减压组低于正常对照组(P<0.01)。结论:神经元继发性损伤是视功能进行性下降的重要原因,视神经减压术有利于减轻视神经间接损伤,较早期(损伤后48h以内)减压可阻止轴突继发性损伤,避免视功能进一步下降,并在一定程度上逆转视功能的损害。     

Effect of training on upper-extremity prosthetic performance and motor learning: a single-case study

Dromerick AW, Schabowsky CN, Holley RJ, Monroe B, Markotic A, Lum P. Effect of training on upper-extremity prosthetic performance and motor learning: a single-case study.

Objectives

To examine the impact of a new prosthesis on an experienced and highly motivated prosthetic limb user, to evaluate the effects of training and the ability of clinical measures to detect change, and to gain insight into the mechanisms by which improvement occurs.

Design

A single-case study.

Setting

An outpatient clinic.

Participant

A bilateral high-arm amputee (right shoulder disarticulation, left above elbow).

Interventions

Provision of new prosthesis and occupational therapy.

Main Outcome Measures

Action Research Arm Test, box and block test of manual dexterity, Jebsen-Taylor Hand Function Test, and speed and accuracy of reaching movements with and without visual guidance.

Results

In this experienced prosthesis user, provision of a new prosthesis led to an immediate worsening in functional limitation. With training, the subject recovered his baseline status and then exceeded it in both proximal and distal function. All study clinical measures detected change, but the change detected varied as much as 300-fold depending on the measure chosen. The clinical improvements were associated with modest improvements in the speed of reaching but not the accuracy of reaching under visual guidance. Improvements in reaching accuracy without visual guidance were seen after 10 trials, suggesting that some motor learning had occurred.

Conclusions

Provision of a new prosthesis can cause functional decline even in an experienced user; this decline can be reversed with training. There is wide variability in sensitivity to change among functional limitation measures. Although some training-related improvements may have been due to increased speed and accuracy of reaching without visual guidance, skill in prosthesis use also plays a role.     

BRAF mutation testing with a rapid,fully integrated molecular diagnostics system

Fast and accurate diagnostic systems are needed for further implementation of precision therapy of BRAF-mutant and other cancers. The novel Idylla^TM BRAF Mutation Test has high sensitivity and shorter turnaround times compared to other methods. We used Idylla to detect BRAF V600 mutations in archived formalin-fixed paraffin-embedded (FFPE) tumor samples and compared these results with those obtained using the cobas 4800 BRAF V600 Mutation Test or MiSeq deep sequencing system and with those obtained by a Clinical Laboratory Improvement Amendments (CLIA)-certified laboratory employing polymerase chain reaction–based sequencing, mass spectrometric detection, or next-generation sequencing. In one set of 60 FFPE tumor samples (15 with BRAF mutations per Idylla), the Idylla and cobas results had an agreement of 97%. Idylla detected BRAF V600 mutations in two additional samples. The Idylla and MiSeq results had 100% concordance. In a separate set of 100 FFPE tumor samples (64 with BRAF mutation per Idylla), the Idylla and CLIA-certified laboratory results demonstrated an agreement of 96% even though the tests were not performed simultaneously and different FFPE blocks had to be used for 9 cases. The Idylla^TM BRAF Mutation Test produced results quickly (sample to results time was about 90 minutes with about 2 minutes of hands on time) and the closed nature of the cartridge eliminates the risk of PCR contamination. In conclusion, our observations demonstrate that the Idylla test is rapid and has high concordance with other routinely used but more complex BRAF mutation–detecting tests.