鸡白痢沙门氏菌鸡胚感染模型建立方法的筛选

为筛选出理想的鸡白痢沙门氏菌宿主感染模型的建立方法,本研究通过选择不同接种方式配合不同菌液浓度接种不同胚龄鸡胚建立感染模型,将90枚SPF鸡胚随机分成9组,每组10枚,分别为A、B、C、D、E、F、G、H、I组,A组为空白对照组,其余各组为模型组,模型组分别以气室接种、蛋壳接触接种的方式对不同胚龄（11和14胚龄）SPF鸡胚接种不同浓度（1×10³、3×10³、9×10³、3×10⁵、9×10⁵ CFU/mL）的鸡白痢沙门氏菌C79-3菌株。每天观察情况直至出雏,待雏鸡孵出后按组分笼饲养,每隔12 h观察1次,连续观察7 d,观察各组雏鸡的临床症状、死亡情况,观察期结束对死亡和存活雏鸡进行剖检、组织病理学检查及鸡白痢沙门氏菌的分离鉴定。结果显示,各模型组均有雏鸡出现明显的鸡白痢病症状并死亡,相较于其他模型组,以11胚龄蛋壳接触感染方式接种9×10⁵ CFU/mL的C79-3菌液的F组和14胚龄气室感染方式接种3×10³ CFU/mL C79-3菌液0.1 mL的H组,鸡胚出壳率较高,可分别达到80%和90%,出壳雏鸡呈现明显鸡白痢病特征,剖检可见肝脏出血、盲肠膨大、卵黄囊吸收不良;组织病理切片可观察到肝脏、盲肠、心脏各有不同程度的损伤,发病率分别100%和88.8%,并且鸡白痢沙门氏菌阳性检出率分别达70%和90%。本研究结果表明,以11胚龄蛋壳接触感染方式接种0.1 mL 9×10⁵ CFU/mL的C79-3菌液和14胚龄以气室感染方式接种0.1 mL 3×10³ CFU/mL的C79-3菌液的两种方法均可用于建立稳定的鸡白痢沙门氏菌宿主感染模型。     

抗旱、耐盐碱、矮秆陆地棉数量性状的遗传相关及主成份分析

对从国内外引进的抗旱、耐盐碱、矮杆棉花资源材料 ,经抗旱、耐盐碱、矮杆性鉴定和比较 ,选取了其中表现较好的 1 9份材料 ,进行了遗传相关和主成份分析 ,结果表明 :在 5 0 mmol· L- 1Na Cland1 0 0 mmol· L- 1Na Cl胁迫下的出苗率间呈正相关 ,抗盐性与产量间呈负相关 ;第一棉铃高度与纤维品质性状呈正相关。抗旱、耐盐碱、矮杆棉花材料的前 6个因子贡献率达 86.5 % ,其分别为第一棉铃高度因子、果枝数因子、单株结铃数因子、矮杆因子、单铃重因子等     

贵州喀斯特高原峡谷和盆地典型区石漠化演变特征分析

以贵州喀斯特高原峡谷和盆地两个典型石漠化综合治理示范区作为研究区,从石漠化演变轨迹和石漠化年变化率两个方面分析石漠化演变特征,结果表明:2000-2013年花江示范区以减弱型为主,并且分布较为集中、成片。红枫湖示范区石漠化以不变型为主,尤其是无石漠化面积不变型占示范区喀斯特面积的42.04%。此外两个示范区都呈现低等级石漠化面积增加,高等级石漠化面积减少的趋势,表明在人为干预下石漠化治理都取得了一定的成效,尤其是花江示范区石漠化治理成果比较显著。     

基于模拟退火算法优化给水管线造价公式研究

给水管线造价公式的精确性对工程经济分析与给水管网优化的科学性和经济性有重大影响。通过五个地区的铸铁管综合单价数据,采用最小二乘法对各组数据进行多项式一至十次的拟合,确定了给水管线造价公式的最佳拟合次数为三次;进一步采用模拟退火算法对拟合的多项式参数进行优化计算,求解给水管线造价公式。结果表明:凭借模拟退火算法随机全局搜索模式以及不受函数性质影响的优势,能够克服传统算法难以求解多阶导数的困难,提高公式拟合精度。     

沼液追肥对制种玉米产量与土壤化学性质的影响

研究沼液作追肥对制种玉米产量及土壤化学性质的影响。文章以制种玉米为试验材料,采用计随机区组试验设,研究沼液追肥对制种玉米经济性状、产量、土壤化学性质、经济效益的影响。结果表明,T1(沼液1500kg·hm-2+142.50 kg N·hm-2)有机质含量最高,与其他处理间达到了显著差异;T1的土壤速效钾含量最高,达12.67 g·kg-1,与CK(150 kg N·hm-2),T2(沼液22500 kg·hm-2+138.75 kg N·hm-2)间达到了显著差异;土壤速效磷含量,土壤p H值,土壤电导率差异不显著;不同处理对穗行数,行粒数、穗粒重的影响差异不显著,T3(沼液30000 kg·hm-2+135 kg N·hm-2)千粒重最高,与CK,T2,T4(沼液37500 kg·hm-2+131.25 kg N·hm-2)间差异达到了显著水平;T2的产量,产值,经济效益最高。沼液做追肥可提高产量,降低生产成本,但必须和化肥配合施用。     

常德市经济型酒店发展的SWOT分析

经济型酒店在全国各地目前的发展是如雨后春笋般崛地而起,发展势头惊人,在大城市得到较好发展后,现在中小城市遍地开花,而因"水土不服"等各方面的因素而发展受挫,出现了一系列混乱竞争、恶性定价等现象。以及各大连锁品牌酒店的入驻,加上本地原有的中、低星级酒店及本土经济型酒店,使得经济型酒店在中小城市的竞争日趋激烈,如何能在新市场中立于不败之地,这是经济型酒店在发展过程中亟待解决的重要问题。此次研究以常德市经济型酒店发展的为例,运用SWOT方法对发展常德市经济型酒店发展的优势、劣势、机遇及威胁进行了分析,并提出常德市经济型酒店发展的对策和措施。     

Study on Expression Difference of IGFBP-5 Gene in Different Tissues of Kazakh and Yanqi Horses

The study aimed to explore the mRNA expression pattern of insulin-like growth factor binding protein-5 (IGFBP-5) gene in different tissues of Kazakh and Yanqi horses.The expression of IGFBP-5 gene in different tissues of heart,liver,spleen,lung,kidney,small intestine,large intestine,cecum,intercostal muscles,longissimus dorsi muscles,brachialis muscle and gluteus in two horses were detected by Real-time quantitative PCR and compared the mRNA expression in the same tissues of two breeds.The results showed that the expression of IGFBP-5 in longissimus dorsi muscle and brachialis muscle of two breeds were significantly higher than other tissues including heart,liver,spleen,lung,kidney,small intestine,large intestine and cecum (P<0.05),and was the lowest in large intestine.The expression of IGFBP-5 in kidney,small intestine,large intestine,cecum,longissimus dorsi muscle and brachialis muscles of Yanqi horse were higher than in the same part of Kazakh horse,and among those in longissimus dorsi muscle and large intestine of Yanqi horse were extremely significantly higher than in the same part of Kazakh horse (P<0.01),and in small intestine and cecum of Yanqi horse were significantly higher than in the same part of Kazakh horse (P<0.05).The test was for further researching the biological function of IGFBP-5 gene,and it could provide a theoretical basis for genetic improvement of production performance of horse in our country.     

The Fluorescent Characterization and Analysis of Two Brucella Attenuated Vaccine Infecting Mouse Macrophagocyte

The experiment was conducted to discuss the difference of binding time of green fluorescent protein B.melitensis M5 (GFP-M5) and B.abortus S19 (GFP-S19) infecting the mouse macrophagocyte (RAW264.7),lysosome,endoplasmic reticulum and golgi body in the initial stage and compare the binding rate of GFP-M5,GFP-S19 with organelle in different timeline,respectively,by confocal laser scanning microscope (CLSM) and flow cytometry.The result showed that GFP-M5 and GFP-S19 were successfully constructed.The intracellular survival ability of Brucella M5,Brucella S19,GFP-M5 and GFP-S19 were not obvisouly affected after infecting RAW264.7.GFP-M5 and GFP-S19 could enter the macrophagocyte in 30 mins,and in 2 h the Brucella could reach lysosome,endoplasmic reticulum and golgi body.In addition,the binding time for two attenuated vaccine did not show differences in 1,2,3 and 4 h.The content of GFP⁺ cell produced by RAW264.7 infected by GFP-M5 and GFP-S19 did not show significant differences (P>0.05).Therefore,the two strains did not have significant differences in the invasion ability in the initial stage of infecting host cell.     

Construction and Identification of Yeast Surface Display Libraries of Ovis aries DRA Gene Exon 2

The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method.