AtmiR156a调控菊苣营养生长与品质

菊苣是南方草地生态畜牧业发展的重要牧草资源,其产量与植物营养生长阶段长短密切相关。本研究采用转基因的方法将拟南芥AtmiR156a在菊苣中过量表达,获得了140株转基因植株,PCR检测阳性率达94.3%。荧光定量PCR分析发现转基因菊苣中AtmiR156a表达量上调7.9倍。AtmiR156a过量表达菊苣与野生型菊苣的发芽率基本相当,但其叶片发生速率显著比野生型快,抽薹时间比野生型推迟20.2 d,开花推迟27.3 d,但是株高比野生型要矮,年均产草量与野生型基本相当。分析第一茬草的品质性状发现AtmiR156a过量表达菊苣叶片中粗蛋白含量比野生型高3.7%,纤维素降低2%,其他品质性状在两个材料中没有显著差异。本研究不仅建立了高效的菊苣遗传转化体系,培育出晚花、速生的菊苣新种质资源,为培育高产耐刈菊苣新品种奠定基础,同时为开展借助其他农作物重要功能基因进行菊苣遗传改良的研究工作提供借鉴,具有重要的理论研究与实际应用价值。     

禽致病性大肠杆菌的分离、O血清群鉴定及药物敏感试验

对2005年5月~2016年3月自河北、山东、河南、江苏、陕西、辽宁等省发病鸡场疑似大肠杆菌病的病死鸡病变部位分离的194株禽致病性大肠杆菌进行了生化特性测定、O抗原血清群鉴定及药敏试验。试验用5种大肠杆菌单因子（O1、O2、O57、O65、O78）阳性血清进行O血清群鉴定,确定了124株O血清群,占鉴定菌株数的63.9%（124/194）,其中O1 3株、O2 26株、O57 15株、O65 47株和O78 33株,O65和O78为优势菌株,而O2和O57血清群菌株次之,O1血清群菌株最少。8种药物（氨苄西林、新霉素、多西环素、磺胺间甲氧嘧啶、替米考星、环丙沙星、头孢噻呋和氟苯尼考）的药敏试验结果表明,受试的5株大肠杆菌分离株均对环丙沙星和新霉素敏感,而对其他药物均有不同程度的耐受。本研究结果表明O65、O78、O2和O57血清群高度流行,为进一步研发多价菌苗奠定了基础。     

逆转录环介导等温扩增技术检测草莓潜隐环斑病毒的研究

采用环介导等温扩增技术（loop-mediated isothermal amplification, LAMP）,建立了草莓潜隐环斑病毒（Strawberry latent ringspot virus, SLRSV）检测方法。根据SLRSV外壳蛋白编码基因上的8个位点,共设计了6条引物,通过逆转录环介导等温扩增得到特征性的瀑布状条带。特异性试验表明,引物对SLRSV的检测具有良好的特异性;灵敏度试验显示RT-LAMP的灵敏度比普通RT-PCR高出100倍。该方法无需特殊的试剂和设备,只需在水浴锅中65 ℃等温扩增,整个检测周期约1.5 h,结果采用SYBR green I染色显示,易于观察和判定。     

苜蓿干草捆贮藏过程中的营养物质含量动态变化研究

为探讨苜蓿干草捆贮藏过程中营养物质含量的动态变化规律,对含水量为15%、打捆密度为150kg/m3的苜蓿草捆在不同贮藏时间(1、10、30、90、180、360d)营养物质含量和相对饲喂价值(RFV)的动态变化情况进行研究。结果表明,在整个贮藏过程中,苜蓿干草的粗蛋白(CP)含量总体呈下降趋势,与贮藏1d时相比,不同贮藏时间段苜蓿干草的CP含量均显著下降(P<0.05);从CP含量角度评价,苜蓿干草等级由1级下降到2级。苜蓿干草的中性洗涤纤维(NDF)含量随贮藏时间的延长而增加,贮藏360d时的NDF含量显著高于贮藏时间在90d之内的NDF含量(P<0.05);从NDF含量角度评价,苜蓿干草等级由2级下降到3级。酸性洗涤纤维(ADF)含量也随贮藏时间的延长而增加,贮藏10d之后ADF含量开始显著增加(P<0.05);从ADF含量角度评价,苜蓿干草等级由1级下降到4级。贮藏时间在30d以上的苜蓿干草DMI值显著低于贮藏时间在30d之内的DMI值(P<0.05);从DMI值角度评价,苜蓿干草等级由1级下降到3级。贮藏时间在90d以上的苜蓿干草DDM值显著低于贮藏时间在90d之内的DDM值(P<0.05);从DDM值角度评价,苜蓿干草等级由1级下降到4级。随着贮藏时间的延长,苜蓿干草的RFV值显著降低(P<0.05),且在贮藏180~360d的时间段内,RFV值下降幅度较大;从RFV值角度评价,苜蓿干草等级由2级下降到3级。综合比较,苜蓿干草在贮藏180d之后营养价值大幅度下降,因此,在贮藏180d之内出售或者饲喂家畜比较适合。     

抗盐酸克伦特罗单链抗体基因的克隆、序列分析及表达

提取分泌抗盐酸克伦特罗单克隆抗体杂交瘤细胞株的总RNA,通过RT-PCR技术,扩增VH和VL,用一段柔性肽链-(G4S)3将VH和VL连接成ScFv。测序后经NCBI Blast分析,所得ScFv具有重组功能性鼠抗体可变区基因的特征。将所得目的基因与pET-22b( )连接,转化E.coli BL21,用IPTG诱导表达,表达蛋白经SDS-PAGE分析,37℃培养5 h表达量较大,25℃诱导表达以可溶性为主。     

猪细小病毒SYBR Green Ⅰ模式实时定量PCR检测方法的建立

根据GenBank猪细小病毒(PPV)的NS1基因序列,设计一对特异性引物,采用SYBR Green Ⅰ随机结合渗入法,建立实时定量PCR检测方法,构建了检测PPV的标准DNA模版,循环阈值(Ct)与标准质粒DNA模板在7.8×101～7.8×108拷贝/μl浓度范围内呈良好的线性关系,相关系数为0.997。该方法用于猪细小病毒的检测具有很高的特异性,其敏感性与常规PCR相比可以提高100倍,可以用于猪细小病毒的快速检测。     

Feline Ilial Fractures: A Prospective Study of Dorsal Plating and Comparison with Lateral Plating

Objective— To (1) report a technique for repair of feline ilial fractures using a dorsally applied bone plate and (2) compare outcome with cats treated by a lateral plate.
Study Design— Prospective study.
Animals— Cats (n=10) with iliac fractures.
Methods— Cats with ilial fractures (January 2005–December 2006) were treated by application of a dorsally applied bone plate. Immediate postoperative radiographs were compared with those taken 4–6 weeks later to assess screw loosening, screw purchase, and pelvic canal narrowing. Owners were contacted for medium-term (>3 month) follow-up. Data were compared with a report of outcome after lateral plating (LP) in 21 cats.
Results— Mean (± SD) screw purchase (89 ± 11 mm) was significantly greater ( P <.01) with a dorsal plate compared with a lateral plate (33 ± 8 mm). Significantly more screws ( P <.01) were used with a dorsal plate (median, 7) compared with a lateral plate (median, 6). Significantly less postoperative pelvic canal narrowing developed in the dorsal plating group between postoperative and 4–6-week follow-up radiography compared with the LP group (2% versus 15%, P <.01).
Conclusion— Dorsal plating of feline ilial fractures results in significantly less screw loosening and pelvic canal narrowing at 4–6 weeks after surgery compared with LP.
Clinical Relevance— Dorsal plating of feline iliac fractures may reduce complications associated with pelvic canal narrowing such as constipation and megacolon.     

Effects of Bacillus subtilis B10 spores on viability and biological functions of murine macrophages

This study was conducted to investigate the effects of Bacillus subtilis B10 spores on the viability and biological functions of murine macrophage. RAW 264.7 cells were stimulated both with and without B. subtilis B10 spores for 12 h. Then cell viability was determined to evaluate the cytotoxic effect of B. subtilis B10 spores to the cells, and the activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH), the production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS), and the secretion of inflammatory cytokines were measured to analyze the functions of macrophages. The results showed that B. subtilis B10 spores were not harmful to RAW 264.7 cells and they also strongly enhanced the activities of ACP and LDH (P < 0.01), remarkably increased NO and iNOS production (P < 0.01), and significantly stimulated the secretion of pro‐inflammatory cytokines, including tumor necrosis factor‐alpha (TNF‐α), interferon‐gamma (IFN‐γ), interleukin‐1 beta (IL‐1β), IL‐6, IL‐8 and IL‐12 (P < 0.01) while they reduced anti‐inflammatory cytokine IL‐10 (P < 0.01). The outcomes suggest that B. subtilis B10 spores are not only safe for murine macrophages, but also can activate these cells and enhance their immune function. The above findings suggest that B. subtilis B10 spores are potentially probiotic.     

不同酶制剂对水稻秸秆和白酒糟混合青贮品质的影响

为研究不同酶制剂对水稻(Oryzasativa L.)秸秆和白酒糟(m︰m=1︰1)混合青贮品质的影响,设置CK(对照)、CEL(250 U·g-1纤维素酶)和XYL(250 U·g-1木聚糖酶)3个处理,青贮120 d后取样测定其青贮品质.结果表明:CEL处理的感官评定等级为良好;CEL和XYL处理的干物质、粗脂肪、...     

若干引进牧草品种的适应性研究

对美国引进的6个牧草种、21个品种进行了适应性和产量比较试验,结果表明:苇状羊茅Festuca arundinacea和冰草Agropyron cristatum的干草总产量较高,其中,苇状羊茅品种Cajun的总产量最高;紫花苜蓿Medicago sativa的干草总产量居于中等水平,在参试的紫花苜蓿品种中,WL323的干草总产量最高;无芒雀麦的产量最低.苇状羊茅、冰草和紫花苜蓿在北京及其周边地区有很好的适应性和推广价值.