重组鸡痘病毒vFV282疫苗株在鸽体内的分布及毒性试验

采用翅内侧皮肤无血管处刺种途径给30日龄幼鸽接种重组鸡痘病毒vFV282疫苗株,利用PCR的方法检测其在鸽体内的分布及其动态并对其毒性进行了研究。结果显示,接种后6 h即在脾脏检测到病毒DNA;接种后1 d,脾、肺PCR检测阳性;3 d,在心、肝、脾、肺、肾、皮肤均检测到病毒DNA;7 d,心、肝、脾、肺、肾、脑PCR检测均呈阳性;10 d,除脑外所有内脏器官中均未检测到病毒DNA,15 d后所有内脏器官PCR检测结果均为阴性。而对照组在整个试验期间PCR检测结果均为阴性。毒性试验表明,重组鸡痘病毒vFV282疫苗株使用安全。     

非洲马瘟疫苗研究进展

本文对非洲马瘟的病毒特性、临床症状和检测方法进行概述,重点对非洲马瘟的传统疫苗和新型疫苗的研究进展进行了详细综述,以期为非洲马瘟疫苗的相关研究提供参考。     

牛分枝杆菌esat-6和mpb70-mpb83基因DNA疫苗的免疫效果

用PCR技术和重叠延伸剪接技术获得牛分枝杆菌esat-6基因和mpb70-mpb83融合基因，连接真核表达载体pcDNA3．1（＋），构建了重组质粒pCE6和pC70-83-E6。分4组免疫小鼠：pCE6组、pC70—83-E6组、pcDNA3．1（＋）和PBS对照组，采用间接EI。ISA法检测免疫小鼠血清特异性抗体水平，MTT法检测免疫小鼠脾淋巴细胞增殖情况和IFN—y分泌情况。结果表明，2重组质粒免疫组小鼠的血清抗体水平持续上升，而2对照组始终维持在较低水平，且pC70-83-E6组小鼠的抗体水平高于pCE6组。经PPD刺激后，pCE6组和pC70—83-E6组小鼠的SI值与2对照组均差异显著（P〈0．05），2重组质粒免疫组间差异不显著（P〉0．05）；2重组质粒免疫小鼠脾细胞产生的IFN—y均显著高于2对照组（P〈0．05），且pC70-83-E6组明显高于其他3组（P〈0．05）。证实本试验构建的2种牛分枝杆菌DNA疫苗可有效诱导实验动物产生体液免疫和细胞免疫应答。     

禽流感油乳佐剂疫苗质量快速检测方法的建立

通过对低温冻融法、研磨法、稀释高速捣碎法、95%乙醇法及三氯甲烷法破乳效果比较,确定三氯甲烷为油乳剂疫苗的破乳剂.将不同抗原含量的H9和H5亚型灭活与非灭活疫苗破乳后分离上层水相,采用红细胞凝集(HA)试验,测定其HA价,结果发现脱乳后上层水相与未做成油乳疫苗并做相应稀释的原液的HA价下降1～3个滴度,且将不同稀释度制备H5亚型疫苗经脱乳处理后上层水相的HA价与荧光定量PCR结果基本一致.所建立的禽流感油乳剂疫苗质量的检测方法具有快速、经济、简便,尤其适合于对大批量及基层应用.     

Vaccination of rainbow trout, Oncorhynchus mykiss (Walbaum), with recombinant and DNA vaccines produced to Flavobacterium psychrophilum heat shock proteins 60 and 70

Flavobacterium psychrophilum heat shock proteins (Hsp) 60 and 70 are highly immunogenic and were therefore investigated as potential vaccine candidates. Recombinant Hsps were purified from Escherichia coli and rainbow trout ( Oncorhynchus mykiss ) were intraperitoneally injected with phosphate buffered saline/Freunds complete adjuvant (FCA), 8 μg of rHsp60/FCA, rHsp70/FCA or a combination of 4 μg each of rHsp60 and rHsp70/FCA. Antibody responses against recombinant Hsp60 and Hsp70 8 weeks post-immunization were observed, but only fish immunized with rHsp70 exhibited highly elevated antibody levels against F. psychrophilum whole cell lysate. Some cross reactivity occurred, which may have been due to the V5 tag common to both proteins. Protection against F. psychrophilum challenge was not observed in any treatments at 8 weeks post-immunization. To further investigate any protective effect of these proteins, hsps were polymerase chain reaction amplified and cloned into pVAX1. Rainbow trout were intramuscularly injected with 8 μg of pVAX1hsp60, pVAX1hsp70 or a combination of 4 μg each of pVAX1hsp60 and pVAX1hsp70. Antibody responses at 4 weeks post-immunization were low and protection was not observed following challenge at 6 or 10 weeks post-immunization. Although Hsps of F. psychrophilum have been shown to be immunodominant, these antigens do not appear to be good vaccine candidates when delivered alone or in combination.     

Surface-exposed expression of Edwardsiella tarda EseB in live attenuated Vibrio anguillarum based on novel surface display systems

Live, attenuated Vibrio anguillarum strains can serve as vectors for the delivery of heterologous antigens for development of multivalent recombinant vaccines. Based on the outer membrane anchoring elements of V. anguillarum , we have previously constructed several efficient surface display systems Lpp-Omporf1, Lpp-OmpU, Lpp-Omp26La, Wza-Omporf1, Wza-OmpU and Wza-Omp26La. In this study, with these constructed surface display systems, a putative antigen protein EseB from pathogenic Edwardsiella tarda was successfully expressed on the surface of an attenuated V. anguillarum strain to get multivalent vaccine candidates. Further immune protection evaluation in zebra fish ( Danio rerio ) demonstrated that the V. anguillarum EseB-display strain AV/pW-26La-B could trigger full protection against V. anguillarum infection and early protection against E. tarda infection in the immunized fish. These results suggest that surface display of heterologous protective antigens in attenuated V. anguillarum could be used as a tool to develop potential V. anguillarum vector vaccine.     

禽霍乱油乳剂灭活苗（TJ株）的研制与应用

从天津地区某些发病鸡场分离的２０余株禽多杀性巴氏杆菌中筛选４株（ＴＪ株）作为制苗用菌种，制成油乳剂灭活苗，进行物理性状检验与安全性，免疫性试验，用实验室制备的禽霍乱油乳剂灭活苗（ＴＪ株）对大港，静海等发病鸡场进行免疫接种，完全控制了禽霍乱的流行。     

猪瘟乳前免疫及其抗体检测

本试验对新生仔猪哺乳前初次进行猪瘟疫苗免疫，至７０日龄时进行第二次免疫，免疫持续至７０日龄和１５０日龄时两次应用Ｄｏｔ－ＥＬＩＳＡ方法进行检测，结果表明，阳性率分别为９２．３％和９１．２５％，说明此免疫方法是一较好的预防猪瘟的免疫程序。     

鸡新城疫CS2实验疫苗的生产及鉴定

应用ＳＰＦ鸡胚生产出３批ＣＳ２实验疫苗，疫苗的病毒价≥１０８．５ＥＬＤ５０／ｍｌ。对２月龄小鸡的最低免疫量（ＭＩＤ）为３～３０ＥＬＤ５０。据此，作者们建议对此种小鸡的实用免疫剂量为１０５ＥＬＤ５０。对这３批疫苗的实验结果显示：１０倍的剂量（即１０６ＥＬＤ５０）对鸡不引起任何不正常现象。免疫持续期达１２个月以上。在２～８℃和－１５℃中分别保存９和２４个月效力不变。     

感染鸡传染性贫血病毒的雏鸡新城疫疫苗免疫后局部体液免疫球蛋白含量变化

本试验对感染鸡传染性贫血病毒（ＣＩＡＶ）雏鸡新城疫（ＮＤ）疫苗免疫及其强毒攻击后，其泪液、气管液、肠液和胆汁的免疫球蛋白ＩｇＧ、ＩｇＭ、ＩｇＡ含量的动态变化进行了检测。结果发现，感染雏鸡泪液的ＩｇＧ、ＩｇＡ和气管液的ＩｇＧ及胆汁的ＩｇＧ、ＩｇＭ、ＩｇＡ含量均于接种ＮＤ疫苗后７～２８天明显低于未感染ＣＩＡＶ的免疫对照雏鸡；泪液的ＩｇＭ和气管液的ＩｇＡ含量在ＮＤ免疫后１４～２８天明显降低；肠液的ＩｇＡ和ＩｇＭ、ＩｇＧ含量分别于免疫后７～１４和７、１４天也显著减少。表明ＣＩＡＶ感染雏鸡对ＮＤ疫苗免疫的局部体液免疫应答能力降低。ＮＤ强毒攻击后，ＣＩＡＶ感染ＮＤ免疫雏鸡泪液、气管液、肠液和胆汁的ＩｇＧ、ＩｇＡ、ＩｇＭ含量及免疫保护率，均明显低于未感染ＣＩＡＶ的免疫对照雏鸡。