猪带绦虫六钩蚴TSOL18基因疫苗表达载体的构建及其免疫原性研究

为研制新型猪囊尾蚴疫苗,将猪带绦虫六钩蚴阶段编码 TSOL18 的基因定向克隆于真核表达质粒 pVAX1, 经筛选、鉴定及 DNA序列分析正确后,将重组质粒 pVAX1/TSOL18 转染 BHK-21 细胞,通过 SDS-PAGE、Western blotting、免疫荧光试验等方法检测细胞中表达的 TSOL18 抗原。结果表明,重组真核质粒 pVAX1/TSOL18 可在 BHK-21 细胞中表达TSOL18 目的蛋白,表达蛋白能被猪囊尾蚴病阳性血清所识别。动物免疫试验表明,真核表达载体能有效地诱导机体产生细胞免疫和体液免疫应答,这为猪囊尾蚴病基因疫苗的研究奠定了良好基础。     

Study on the Change Rule of Egg Yolk Antibody in Laying Hens after First Immunization by Japanese Encephalitis Virus

The purpose of this experiment was to study the immunization rule of the egg yolk antibody affected by different vaccines,immunization dose and injection ways and further to discuss the optimal immunization procedures of the laying hens for the preparation of egg yolk antibody against swine Japanese encephalitis virus.180 brown laying hens without any vaccines were selected and divided into 18 groups randomly,each group of 10 hens.Groups 1,2 were the control groups,injected with the sterile saline;Groups 3 to 10 were injected with subcutaneous or intramuscular injection,and the vaccine was injected with 0.2,0.5,1.0 and 1.5 mL successively.Groups 11 to 18 were also adopted two kinds of injection,followed by the same dose of vaccine immunization.Six eggs of each experimental group were gathered before immune day and after 3,7,10,14,18,21 and 28 days,the egg yolk antibody was extracted and the titer was determined.As a result,the egg yolk antibody titers of groups 1 to 6,11 and 12 were all 0,and no significant immune response produced;The hens from 7 to 10 groups were injected with the inactivated vaccine.After 7 days,the average antibody titer reached the peak,and the duration of the antibody was 14 days.The hens from 13 to 18 groups were injected with the attenuated virus vaccine.After 14 days,the average antibody titer reached the highest value,and the duration of the antibody was 21 days.The egg yolk antibody titers were not significantly different in the two compared experiment groups with the same injection dose but with different injection ways (P>0.05).With the same injection way of each experiment group,and the difference was significant (P>0.05).Compared with some groups with the same injection and vaccine,the titer of yolk antibody was gradually increased with the increase of the immune dose,and the difference was significant (P<0.05).The results showed that,no matter intramuscular or subcutaneous injection,in order to produce a significant immune response to hens,the immune antigen dose was 1.0 mL inactivated vaccine or 0.5 mL attenuated vaccine at least.Compared with the attenuated and inactivated vaccine,inactivated vaccine stimulated the body to produce the antibody faster,but the maintenance time was shorter;The lower dose of attenuated vaccine could stimulate the body to produce antibodies,but the speed was slower,the maintenance time was longer.     

恩诺沙星混悬液在猪体内残留消除规律研究

采用高效液相色谱法研究恩诺沙星(口服)混悬液在猪体内各组织中的残留消除规律.恩诺沙星(口服)混悬液,按每头猪10 mg/kg体重的剂量灌服给药,连续使用3 d之后,宰杀猪,取组织.组织样品经磷酸盐缓冲液提取,C18固相萃取柱净化,过膜,用流动相0.05 mol/L磷酸溶液/三乙胺一乙腈(82+18)溶解,微孔过滤,进行...     

3种兔球虫18S rDNA部分序列测定与系统发育分析

采用单卵囊分离法从河北某兔场分离大型艾美耳球虫、黄艾美耳球虫及肠艾美耳球虫,接种无球虫兔后获得大量纯种卵囊,CTAB法提取孢子化卵囊基因组DNA。利用艾美耳属球虫18S rDNA保守引物,PCR扩增3种兔球虫18S rDNA片段,产物纯化后测序。将3种球虫18S rDNA测序结果与GenBank中发布的兔球虫18S rDNA序列用DNAStar软件进行比对。使用MEGA4.0软件对兔球虫18S rDNA进行同源性比较,并绘制遗传进化树。结果表明,大型艾美耳球虫扩增出大小为1 521bp的18S rDNA片段;黄艾美耳球虫及肠艾美耳球虫均扩增出大小为1 520bp的18S rDNA片段。序列比对结果显示,3种河北株兔球虫与GenBank中相应的3种兔球虫18S rD-NA(EF694016、EF694011、EF694012)相似性分别为99.6%、99.6%和100%。3种河北株兔球虫序列和GenBank中兔球虫18S rDNA序列(EF694007-EF694017)位于一个单系集群。     

仔犬主要感觉行为发育的观察

犬的感觉行为是犬最基本的行为构成,其发育是一个连续的过程.了解和掌握仔犬期(0-45日龄)主要感觉行为的发育特征,对提高仔犬感觉统合能力、行为发育、仔犬期的饲养管理水平和训练水平具有重要意义.     

无磷酸氢钙日粮对生长猪的影响研究

将48头杜×长×大三元杂交生长猪随机分为两组,分别饲喂正常磷水平日粮和无磷酸氢钙日粮,结果表明与正常磷水平日粮相比,无磷酸氢钙日粮可显著提高磷的表观消化率(P＜0.05),增加猪场的纯收入,减少粪中磷的排放以保护猪场周边的生态环境.     

禽蛋中农药残留检测方法研究进展

禽蛋是人类获取动物蛋白的重要来源之一,禽蛋中残留的农药导致食品安全隐患,威胁人类健康,因此迫切需要建立有效的检测方法,为禽蛋产品的质量安全提供技术保障.文章综述了近十年来国内外文献报道及我国国家标准中规定的禽蛋样品农药残留的前处理方法和分析检测方法,为进一步研究建立简便高效的分析检测新方法、新技术提供参考.     

溶氧因素对冬虫夏草摇瓶培养的影响

对冬虫夏草菌丝体摇瓶培养中影响溶氧的主要因素进行分析。通过测定不同摇床转速、装液量、封口方式下摇瓶培养后的菌丝得率,并对其进行单因素分析,选择单因素较优水平,然后进行3因素正交试验。结果表明,最适宜溶氧条件为摇床转速140r/min、装液量50%、封口采用无菌培养容器封口膜;各因素影响培养的显著性依次为摇床转速、装液量、封口方式;不同摇床转速下对有效成分含量测定显示,摇床转速对虫草多糖及虫草酸含量影响不显著。     

水分对柠条萌蘖株和未平茬株光合参数及调渗物质的影响

对柠条Caragana korshinskii平茬后的萌蘖株和未平茬的对照株在夏季旺盛生长期进行了水分处理,并研究了浇水前后萌蘖株和对照株光合参数及调渗物质的变化。浇水前,萌蘖株具有较高的水势 (LWP)、光合速率(A)、气孔导度(G)、蒸腾速率(E)、光系统II最大光化学量子效率(Fv/Fm)和光系统II实际量子产率(△F/Fm′);而对照株相应指标较低,但对照株光化学猝灭系数（NPQ）,脯氨酸(Pro)、 丙二醛(MDA)含量较高。浇水后,对照株LWP、A、G、E、Fv/Fm、△F/Fm′均有不同程度的上升,而NPQ值和Pro、MAD的含量下降。浇水对萌蘖株和对照株叶绿素(Chla+b)含量影响不显著,且浇水前后萌蘖株均高于对照株。结果表明：萌蘖株在生长季所受干旱胁迫较小,植物将更多的光能以光合作用的形式利用,植物生长迅速;而对照株所受干旱胁迫的影响较为严重,光合速率低,植物将更多的光能以热的形式消耗,导致植物生长缓慢。