Zhang Litai, Huang Xiaofeng, Xue Bai, Peng Quanhui, Wang Zhisheng, Yan Tianhai, Wang Lizhi
Institute of animal nutrition, Sichuan Agricultural University, Yaan, Sichuan, China.
Agri-Food and Biosciences Institute, Hillsborough, United Kingdom.
PLoS One. 2015 Oct 7;10(10):e0140086. doi: 10.1371/journal.pone.0140086. eCollection 2015.
Vaccination through recombinant proteins against rumen methanogenesis provides a mitigation approach to reduce enteric methane (CH4) emissions in ruminants. The objective of present study was to evaluate the in vivo efficacy of a new vaccine candidate protein (EhaF) on methanogenesis and microbial population in the rumen of goats. We amplified the gene mru 1407 encoding protein EhaF using fresh rumen fluid samples of mature goats and successfully expressed recombinant protein (EhaF) in Escherichia coli Rosetta. This product was evaluated using 12 mature goats with half for control and other half injected with 400ug/goat the purified recombinant protein in day 1 and two subsequent booster immunizations in day 35 and 49. All measurements were undertaken from 63 to 68 days after the initial vaccination, with CH4 emissions determined using respiration calorimeter chambers. The results showed that the vaccination caused intensive immune responses in serum and saliva, although it had no significant effect on total enteric CH4 emissions and methanogen population in the rumen, when compared with the control goats. However, the vaccination altered the composition of rumen bacteria, especially the abundance of main phylum Firmicutes and genus Prevotella. The results indicate that protein EhaF might not be an effective vaccine to reduce enteric CH4 emissions but our vaccine have potential to influence the rumen ecosystem of goats.
通过重组蛋白进行疫苗接种以对抗瘤胃甲烷生成,为减少反刍动物肠道甲烷(CH4)排放提供了一种缓解方法。本研究的目的是评估一种新型候选疫苗蛋白(EhaF)对山羊瘤胃甲烷生成和微生物种群的体内功效。我们使用成年山羊的新鲜瘤胃液样本扩增了编码蛋白EhaF的基因mru 1407,并在大肠杆菌Rosetta中成功表达了重组蛋白(EhaF)。使用12只成年山羊对该产品进行评估,一半作为对照,另一半在第1天注射400μg/只的纯化重组蛋白,并在第35天和第49天进行两次后续加强免疫。所有测量均在初次接种疫苗后的63至68天进行,使用呼吸热量计室测定CH4排放。结果表明,与对照山羊相比,接种疫苗后血清和唾液中产生了强烈的免疫反应,尽管对肠道总CH4排放和瘤胃中产甲烷菌数量没有显著影响。然而,接种疫苗改变了瘤胃细菌的组成,尤其是主要菌门厚壁菌门和普雷沃氏菌属的丰度。结果表明,蛋白EhaF可能不是一种减少肠道CH4排放的有效疫苗,但我们的疫苗有潜力影响山羊的瘤胃生态系统。