Identification of lactic acid bacteria in the feces of dairy cows fed whole crop maize silage to assess the survival of silage bacteria in the gut

In order to assess the survival of lactic acid bacteria (LAB) in whole crop maize silage in the gut of dairy cows, one representative silage sample and three different feces samples were collected from dairy cows on three dairy farms in Hua Bei, China and three dairy farms in Kyushu, Japan. The composition of the bacterial community was examined by denaturing gradient gel electrophoresis and quantitative polymerase chain reaction. Lactobacillus acetotolerans was detected in all bunker‐made maize silage samples, regardless of the dairy farm or sampling region from which they were sourced. A total of eight LAB species were detected in the maize silage samples, of which three (L. acetotolerans, L. pontis and L. casei) appeared to survive digestion. The populations of L. acetotolerans in silage and feces were 10^6–7 and 10^3–4 copies/g, respectively, indicating that, even for the LAB species showing potential survival in the gut, competition in this niche may be harsh and the population may substantially decrease during the digestion process. It may be difficult for silage LAB to survive in the gut of silage‐fed dairy cows, because marked decrease in population can take place during the digestion process, even for surviving species.     

2006—2012年鲁豫冀地区15株猪繁殖与呼吸综合征病毒的分离鉴定及ORF5/Nsp2基因的遗传变异分析

为研究鲁豫冀地区猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)的遗传变异情况,对2006—2012年来自3省区发病猪场的42份样品进行PRRSV分离鉴定,并进行了生物学特性研究和PCR鉴定,结果显示先后分离到15株PRRSV。分别采用RT-PCR扩增其ORF5基因和部分Nsp2基因并测序,与GenBank中68个ORF5序列和40个Nsp2序列的推导氨基酸序列进行比对,遗传变异分析结果表明,15株分离株均属于美洲型毒株,其中13个毒株Nsp2基因推导的氨基酸序列均存在氨基酸的不连续缺失,其ORF5基因推导的氨基酸序列与JXA1株有较高的同源性(95.5%~97.5%);SDDY2007株与疫苗株RespPRRS MLV和VR2332株亲缘关系相近,处于同一个亚群中;而HN25-2009分离株Nsp2基因推导的氨基酸序列有30个氨基酸的特征性缺失,其ORF5基因推导的氨基酸序列的遗传进化分析结果显示该分离株处于VR2332所在亚群(氨基酸同源性97.5%),具有一定特殊性。本试验结果表明,2006—2012年高致病性PRRSV是鲁豫冀地区的优势流行毒株,且存在疫苗毒株,3省区流行毒株间有一定遗传差异,但无明显地域特征。     

猪骨形态发生蛋白-6基因多态性及其与产仔性能的关联性分析

试验旨在研究猪骨形态发生蛋白-6（bone morphogenetic protein 6, BMP-6）基因多态性及其与产仔性能的关联。采用PCR-SSCP技术检测了野猪、民猪、长白猪、北京黑猪、法系大白猪5个猪种268个个体BMP-6基因的多态性,并分析了BMP-6基因多态性与法系大白猪产仔性能的关联性。结果表明,猪BMP-6基因CDS区1104位存在1个G→A的同义突变,产生AA、AB、BB 3种基因型,其中在野猪和民猪中,A等位基因频率高;在长白猪、北京黑猪和法系大白猪中,B等位基因频率高。不同基因型对法系大白猪产仔数和产活仔数的影响差异显著（P<0.05）,初步认为BMP-6基因多态性与产仔性能显著关联。     

Effect of three insect-resistant maizes expressing Cry1Ie,Cry1Ab/Cry2Aj and Cry1Ab on the growth and development of armyworm Mythimna separata(Walker)

Three transgenic maize events(IE09 S034, Shuangkang 12–5 and C0030.3.5) produced Cry1 Ie, Cry1 Ab/Cry2 Aj and G10-EPSPS, Cry1 Ab and EPSPS, respectively, all of which target the Asian corn borer. The oriental armyworm Mythimna separata(Walker) is the secondary target. In this study, the effects of the three Bt maizes on the development and survival of armyworm were studied. The results showed that IE09 S034 had insecticidal activity against 1 st instar larvae, and the survival rate of armyworm fed with Bt maize for 10 days was 46.2%, significantly lower than that of the control. The larvae at 3 rd–6 th instar were more tolerant of the Bt toxin than the early instar larvae. However, Shuangkang 12-5 had good insecticidal activity against 1 st–5 th instar larvae. The mortality was nearly 100% when the larvae were fed with Shuangkang 12-5 before 3 rd instar, and the toxin had quick-acting efficacy. This event significantly inhibited the development of armyworm; that is, the larval duration of the 3 rd and 4 th instar larvae fed with Shuangkang 12-5 was prolonged by 4.5 and 3.0 days, respectively. The pupal weight and egg number were also significantly lower than those of the control. For C0030.3.5, it could control 1 st–5 th instar larvae effectively. The mortality rates were all over 50% if 1 st–3 rd larvae were fed with this event. The pupal weight of 4 th–6 th instar larvae fed with Bt maize were only 53.9, 56.8 and 54.6%, respectively, compared to that of the control. The number of eggs laid was significantly less than the control. The results indicate that all three transgenic maize events exhibit the potential to provide effective control of early instar larvae of armyworm, which can be commercialized in future to control lepidoptera pests such as Asian corn borer and armyworm.     

7个鲜食葡萄生长发育特性观测

【目的】 研究新疆南疆地区主栽鲜食葡萄品种生长发育特性,为果园栽培管理提高品质提供理论依据。【方法】 以新疆阿图什市相同立地条件下的7个鲜食葡萄品种为材料,研究结果习性、物候期、叶片性状、开花特性及果实发育动态变化。【结果】 各葡萄品种萌芽率79%~93%,结果系数基本一致,但结果枝率差异明显,无核白鸡心最高为1.21,梦幻无核、白木纳格和和田红最低为1.03;花期持续6~11 d;各葡萄品种叶片的差异程度分别为比叶重>单叶重量>叶面积>叶长>叶柄长>叶长宽比>叶柄宽>叶宽>叶柄厚,比叶重新郁最高0.08,白木纳格最小0.03。【结论】 葡萄叶片叶绿素含量最高的是白木纳格葡萄;葡萄果实生长发育动态呈现双“S”曲线和单“S”曲线。     

褪黑素浸种对盐碱胁迫下大豆种子萌发的影响

为研究褪黑素浸种对盐碱胁迫下大豆种子萌发的影响,以大豆品种垦丰16为试验材料,研究100μmol/L褪黑素浸种对不同浓度混合盐碱（NaCl和NaHCO₃物质的量按1:1混合,配制为0、25、50、100、200mmol/L的盐碱溶液）胁迫下大豆种子萌发的影响。结果表明：与对照相比,随盐碱浓度提升,大豆种子发芽率、发芽指数、芽鲜重和芽干重呈不断降低趋势,超氧化物歧化酶、过氧化物酶、过氧化氢酶、谷胱甘肽还原酶和谷胱甘肽过氧化物酶活性呈现先升高后降低的趋势,抗坏血酸过氧化物酶活性呈不断上升趋势;渗透调节物质可溶性糖、可溶性蛋白和脯氨酸含量呈现先升高后降低的趋势;膜脂过氧化产物丙二醛、H₂O₂含量、$\mathop{{O}}_{2}^{{\mathop{}_{\ .}^{-}}}$产生速率呈不断上升趋势。褪黑素浸种可提高不同浓度盐碱胁迫下大豆种子发芽率、发芽指数、芽鲜重、芽干重和抗氧化酶活性以及渗透调节物质含量,降低丙二醛、H₂O₂含量和$\mathop{{O}}_{2}^{{\mathop{}_{\ .}^{-}}}$产生速率。综上所述,褪黑素浸种可以缓解不同浓度盐碱胁迫对大豆种子发芽的抑制作用,并能提高抗氧化酶活性和渗透调节物质含量,降低膜脂过氧化程度。     

递增注射脂多糖诱导慢性免疫应激对仔猪生长性能的影响

本试验旨在研究递增30%注射脂多糖(LPS)诱导仔猪慢性免疫应激的效果及对仔猪生长性能、营养物质表观消化率的影响。选取16头断奶仔猪,随机分为2组(对照组、LPS组),单笼饲养。LPS组隔天静脉注射LPS,初始注射剂量80μg/kg BW,随后逐次递增30%,对照组注射相应剂量的生理盐水,试验期15 d。研究发现:仔猪注射LPS第1、5天血浆白细胞介素-1β和皮质醇含量极显著高于对照组(P0.01),第9天血浆皮质醇有高于对照组的趋势(P=0.10),整个试验期内血浆结合珠蛋白含量显著高于对照组(P0.05)。与对照组相比,注射LPS仔猪在试验的3个阶段(1~4 d、5~8 d和9~15 d)平均日增重分别降低了40.00%(P0.01)、29.52%(P0.05)和19.30%(P0.05),平均日采食量分别降低了25.09%(P=0.01)、23.15%(P0.05)和19.47%(P0.05),增重耗料比与对照组无显著差异(P0.05)。与对照组相比,注射LPS仔猪饲粮干物质表观消化率下降了5.43%(P0.05),能量表观消化率有降低的趋势(P=0.06),粗脂肪和粗蛋白质表观消化率略有降低(P0.10)。试验结果表明,递增30%注射LPS成功诱导仔猪慢性免疫应激,降低了仔猪平均日采食量和营养物质表观消化率,抑制了仔猪生长。     

牛病毒性腹泻病毒单克隆抗体捕获ELISA方法的建立

以纯化的牛病毒性腹泻病毒(BVDV)作为抗原免疫BALB/C系小鼠,通过细胞融合和克隆筛选出稳定分泌BVDV抗体的杂交瘤细胞株,将杂交瘤细胞接种于BALB/C系小鼠的腹腔内诱生腹水,腹水单抗与BVDV均呈阳性反应.将5A9、2G3和5C2(分别结合不同的抗原结合位点)诱生的BVDV单抗纯化后等量混合包被酶标板,与鸡抗BVDV IgY(检测抗体)联合应用,建立BVDV抗原捕捉ELISA(AC-ELISA)方法.采用方阵滴定法确定单抗、鸡抗BVDV IgY的最适工作浓度,用阴性组织样品作为判定检测结果的D490临界值,最后以建立的AC-ELISA检测临床粪样棉拭子27份,结果表明:该方法敏感性、特异性高,与全血病毒分离结果的符合率迭86.36%;与全血和粪样RT-PCR检测结果的符合率分别为89.47%和94.74%.阴性对照RT-PCR、病毒分离和AC-ELISA检测均为阴性.     

T-RFLP技术及其在动物胃肠道微生物群落多样性研究中的应用

随着分子生物学技术的飞速发展,越来越多的新兴研究手段应用于胃肠道微生态系统的研究之中。介绍了末端限制性片段长度多态性技术(T-RFLP)的基本原理、操作流程和优缺点,并总结了该技术在动物胃肠道微生态研究中的应用现状。     

复合微生物制剂对重口裂腹鱼生长、消化酶活性、肠道菌群及水质指标的影响

本试验旨在研究复合微生物制剂对重口裂腹鱼生长、肠道消化酶和菌群以及水质指标的影响.选取健康平均体重(250.00±0.50)g的重口裂腹鱼180尾,随机分成2组(对照组和试验组),每组3个重复,每个重复30尾,对照组饲喂基础日粮,试验组饲喂在基础日粮中添加2‰复合微生物制剂的试验日粮,试验期120 d.结果显示:与对照组相比,饲喂复合微生物制剂可提高重口裂腹鱼的特定生长率和饲料效率,且随养殖时间的增加作用效果越明显;可显著提高前肠、中肠和后肠的蛋白酶、脂肪酶、淀粉酶活性(P<0.01);可显著或极显著增加前、中、后肠芽孢杆菌、双歧杆菌和乳杆菌的菌群数量(P<0.01或P<0.05);可显著或极显著降低试验中、后期水体中活性磷酸盐和氨氮浓度(P<0.01或P<0.05).由结果可知,复合微生物制剂能改善水质,提高重口裂腹鱼肠道消化酶活性、增加肠道有益菌群数(芽孢杆菌、双歧杆菌和乳杆菌),促进鱼体生长.