瘤胃微生物在木质纤维素价值化利用的研究进展

瘤胃是反刍动物消化的第一个腔室，各种微生物（细菌、真菌和原生动物）相互作用将木质纤维素植物生物降解为易于代谢的化合物。瘤胃也是目前自然界公认的木质纤维素高效降解和利用的天然反应器，其真菌和细菌可分泌多种木质纤维素降解酶，在木质纤维素生产生物燃料和化学用品方面具有潜在的价值。因此，本研究在瘤胃内木质纤维降解的微生物及其降解木质纤维素相关酶的基础上，重点综述了瘤胃微生物在木质纤维素生物转化为乙醇、生物化学（有机酸）及沼气等方面的研究进展，旨在为瘤胃微生物和瘤胃酶在木质纤维素价值化利用方面的研究和应用提供新的方法和思路。     

莫能菌素浓度对混合瘤胃微生物体外发酵的影响

本试验研究了莫能菌素对混合瘤胃微生物体外发酵的影响 ,以期确定莫能菌素在瘤胃内容物中的适宜添加浓度。混合瘤胃微生物由饲喂花生秧基础日粮的阉公牛瘤胃内容物和瘤胃液制备 ,用简易人工瘤胃装置测定莫能菌素浓度对产气量、瘤胃乙丙酸比例、粗纤维降解率和真蛋白产量的影响。结果表明 ,莫能菌素浓度强烈地抑制产气量 ,两者呈线性负相关(P<0.05);粗纤维降解率开始时随莫能菌素浓度的升高而增加 ,在浓度为0.078mg/L时达到最大值 ,两者呈二次幂函数关系 ;添加莫能菌素降低了瘤胃乙丙酸比例 ,但瘤胃乙丙酸比例随莫能菌素浓度的提高有逐渐增加的趋势 ;在莫能菌素浓度为0.1～2.0mg/L的范围内 ,添加莫能菌素对蛋白质的产量没有明显影响     

大豆和百脉根古老原核基因的全基因组鉴定与比较分析

在漫长的进化过程中,高等真核生物从原核生物中获取了大量基因,对这些基因进行鉴定分析,可为研究高等植物系统演化及基因组水平比较分析提供理论依据。为了进一步解析大豆和百脉根古老原核基因在其基因组中所起的作用及进化关系,本研究利用生物信息学技术对大豆和百脉根基因组古老原核基因进行全基因组鉴定,并对其特征及功能进行了比较分析。研究结果表明,大豆(40.6%)古老原核基因的占比高于百脉根(33.9%),且多分布于内共生细胞器线粒体、叶绿体中。此外,古老原核蛋白质的结构域在大豆和百脉根基因组中的分布相似,表明其在大豆和百脉根进化过程中具有较高的共线性和保守性。通过GO注释发现大豆与百脉根中古老原核蛋白主要分布在膜系统、细胞、细胞组分中。在分子功能上,大豆古老原核蛋白更多地参与代谢和发育过程,这可能与其接受了更多的人为选育有关。在生物过程中,古老原核蛋白更多参与催化反应和结合反应,主要以酶的形式在发挥作用。通过比较分析,本研究揭示了古老原核蛋白在大豆和百脉根中进化的独特模式,可为其他豆科植物基因组分析提供一定理论基础。     

冷季放牧补饲对甘肃高山细毛后备母羊瘤胃代谢参数及瘤胃微生物数量的影响

旨在筛选出甘肃高山细毛羊后备母羊科学合理的补饲料,改变牧区由于营养供给不足导致甘肃高山细毛羊2.5岁才能配种的饲养方式。试验选择甘肃高山细毛羊(12月龄)48只,随机分为试验a(1号料)、b(2号料)、c(3号料)组和对照组(不补饲),进行90 d放牧补饲试验。结果表明,1)瘤胃pH试验a组和b组差异极显著(P<0.01)、c组显著(P<0.05)低于对照组,瘤胃氨态氮、蛋白氮、总氮浓度试验组均极显著高于对照组(P<0.01),总挥发性脂肪酸(VFA)试验a组和b组均极显著高于对照组(P<0.01),b组显著高于c组(P<0.05);乙酸和丙酸摩尔比a组极显著高于对照组(P<0.01),b和c组显著高于对照组(P<0.05);丁酸摩尔比a组极显著高于c组(P<0.01);乙/丙b组显著低于对照组(P<0.05)。2)瘤胃纤维降解菌和原虫数目试验组均极显著(P<0.01)或显著(P<0.05)高于对照组。综上所述,补饲均能正向影响甘肃高山细毛羊后备母羊瘤胃代谢参数及瘤胃微生物数量,以1号料补饲效果最优,2号料也能起到理想效果。     

Impact of cold-season grazing and supplementary feeding on rumen metabolic parameters and micro-organisms in the ewes of Gansu Alpine Fine Wool Sheep北大核心CSCD

An experiment was conducted to scientifically identify the appropriate feed for Gansu Alpine Fine Wool ewes in order to change the animals' breeding mode, which currently can only be mated at the age of 2.5 years due to insufficient nutritional supply. The experiment selected 48 12-month-old ewes and randomly divided them into group a (Diet 1), group b (Diet 2), group c (Diet 3), and a control group with no supplementary feeding. The feed test was run for 90 days. The rumen pH of groups a and b were very significantly higher (P<0.01) or group c significantly lower (P<0.05) than control group. The rumen ammonia nitrogen, protein nitrogen and total nitrogen of groups a, b and c were all higher than control group (P<0.01). The total volatile fatty acids of group a and b were all higher than the control group (P<0.01), group b significantly higher than group c (P<0.05). The molar ratio of acetic acid and propionic acid in group a was very significantly higher than control group (P<0.01), group b and c were significantly higher than control group (P<0.05). The molar ratio of butyric acid in group a was very significantly higher than group c (P<0.01). The ratio of acetic acid and propionic acid in group b was significantly lower than control group (P<0.05). The number of several rumen fiber-degrading bacteria and protozoa in groups a, b and c were very significantly (P<0.01) or significantly (P<0.05) higher than the control. In conclusion, this study shows that supplementary feeding can positively affect the rumen metabolic parameters and the number of several rumen microorganisms in Gansu Alpine Fine Wool ewes. In general, the effects of Diet 1 were the strongest, while Diet 2 also produced notable improvements. © 2018, Editorial Office of Acta Prataculturae Sinica. All rights reserved.     

Research Progress on Bacterial Resistance in Environment

Antibiotic resistance genes have become a recognized environmental pollutant, threatening the ecological safety and human health. The application of antibiotics in the clinical and animals breeding, making the environmental microorganisms living with the impact of the residue of antibiotics and elements of resistance genetic and leading to antibiotic resistant bacteria to gain a competitive advantage and destroyed the stability of ecosystem. In this paper, we expounded the concept of resistance gene transmission by the view of macro environment. Through analyzing the mechanism of resistance development,environmental pollution caused by drug resistance and the impact of environmental microorganism for drug resistance, we clarified the key role of the environment in the development of the characteristics of bacterial resistance and analysis environmental resistance.Such as the ecological diversity of flora, the types and the spread of resistant bacteria, the residues of antibiotics and the transmission of the resistance genes.     

Expression,Purification and Activity Evaluation of Mb1230 Gene of Mycobacterium bovis

To explore the value of Mb1230 protein of Mycobacterium bovis in the diagnosis of bovine tuberculosis,we obtained the Mb1230 gene by PCR and constructed recombinant plasmid pET-22b-Mb1230.Recombinant Mb1230 protein was obtained by IPTG induction and purified by affinity chromatography.The activity of the recombinant protein was evaluated by TST test,IGRA test and indirect ELISA.The size of the recombinant protein matched with the theoretical value proved by SDS-PAGE;Western blotting result showed that the recombinant protein could react with mouse anti-His antibody,and had specific band;The results of TST test,IGRA test and indirect ELISA test also showed the recombinant protein had antigenic activity.The results indicated the recombinant protein Mb1230 had good B cell and T cell activity,so,it had the potential application in the diagnosis of bovine tuberculosis.     

Sequence Analysis and Prokaryotic Expression of NSP1 Gene of Bovine Rotavirus UK Strain

Rotavirus is a major cause of acute diarrhea in both many kinds of young animals and children under 5 years old.Rotavirus NSP1, a 55 ku RNA binding protein, is the product of gene 5, which can subvert innate immune responses and be one of virulent determinant factors.According to the sequence in GenBank, specific primers targeting to NSP1 gene were designed and the gene was amplified by RT-PCR, following by being cloned into the pET-28a(+) vector.It showed that the full length of NSP1 gene was 1 473 bp, encoding 491 amino acids.The NSP1 shared the highest identity with WC3 strain.The recombinant protein was induced in E.coli Rosetta(DE3) by IPTG and was analyzed by SDS-PAGE and Western blotting.The results revealed that NSP1 recombinant protein existed in the form of inclusion body with the molecular weight of 55 ku.The purified recombinant protein could be recognized by His-tag antibody.This study laid the foundation for further research on the relationship between the intracytoplasmic location of NSP1 protein and its activity.     

传染性法氏囊病病毒主要结构蛋白VP2、VP1和VP2-VP1串联基因的原核表达及其初步应用

为获得传染性法氏囊病病毒(IBDV)特异性抗体检测用抗原VP2、VP1及VP2-VP1蛋白,分别设计引物扩增IBDV野毒株NN1172的VP2和VP1基因,并扩增VP2和VP1基因中抗原性和亲水性较好的重要区域,通过PCR扩增基因串联方法对截短的VP2和截短的VP1基因进行串联,首次获得VP2-VP1串联基因,并对VP2、VP1和VP2-VP1串联基因进行了原核表达和鉴定。结果成功构建了原核表达载体pET-VP2、pET-VP1和pET-VP2-VP1;诱导表达条件显示,3个重组质粒分别转入BL21菌株后经0.05 mmol/L IPTG诱导表达,分别得到分子量为69、114和63 kDa的VP2、VP1和VP2-VP1重组蛋白,且均以包涵体形式表达,3个重组蛋白分别于诱导后5、3和6 h时表达量最多。Western blot结果显示,表达的VP2、VP1和VP2-VP1蛋白与鸡抗IBDV阳性血清均具有良好的反应原性。以纯化的VP2、VP1和VP2-VP1蛋白作为包被抗原对传染性支气管炎病毒(IBV)、呼肠孤病毒(ReoV)、禽白血病病毒(ALV)和新城疫病毒(NDV)4种阳性血清检测均为阴性,表明所获得的纯化蛋白具有高度的特异性;对免疫了IBD灭活疫苗,IBD基因工程疫苗和IBD弱毒疫苗的商业鸡群进行抗体检测,结果均能显示疫苗免疫后机体抗体水平的变化趋势。本研究表明利用该原核表达系统所表达的3个蛋白均具有良好的免疫反应活性,为IBDV特异性抗体的检测和新型亚单位疫苗的研发奠定基础。