山羊痘灭活疫苗免疫持续期试验

将实验室制备的3批山羊痘油佐剂灭活疫苗,分别免疫接种一定数量无羊痘抗体的山羊,山羊于免疫接种后4.5个月、7个月、9.5个月和12个月用AV40株强毒进行攻击。结果显示,3批疫苗接种的山羊免后4.5个月攻毒均100%保护,免后7个月攻毒平均80%保护,免后9.5个月攻毒平均60%保护。实验表明,此山羊痘灭活疫苗免疫后6个月能够维持较高的抗体水平,其免疫持续期可规定为6个月。     

新城疫F－Ⅶ型－－鹅源禽副粘病毒灭活疫苗的研制与免疫效力试验

用GPV-SF02株作为抗原液,经福尔马林灭活后,与油佐剂混合,制成SF02灭活疫苗。同时将ND灭活疫苗、ND弱毒疫苗和SF02灭活疫苗按不同剂量分别免疫川白鹅和AA鸡,结果用1羽份SF02灭活疫苗免疫的鹅与鸡,能100%抵抗SF02株和F48E9株的攻击;用1羽份ND灭活疫苗免疫的鸡能100%抵抗F48E9株的攻击,而免疫的鹅对F48E9株的抵抗力为60~80%;用1羽份ND弱毒疫苗免疫的鹅,其保护率在0~50%,免疫的鸡则有100%的保护率。     

牛传染性鼻气管炎病毒LN01/08株灭活疫苗的研制

为研制牛传染性鼻气管炎病毒(IBRV)灭活疫苗,本研究以国内分离鉴定的IBRV LN01/08株为种毒,优化病毒增殖条件,获得病毒滴度达108.0TCID50/mL,将其灭活制备疫苗.为比较不同免疫佐剂的效果,分别以矿物质白油和Montanide ISA206佐剂配制灭活疫苗,进行牛体免疫试验.临床观察和血清中和抗体检测结果表明,Montanide ISA206佐剂乳化的疫苗在降低副反应和增强免疫效果方面优于矿物质白油佐剂.应用Montanide ISA206佐剂制备3批灭活疫苗,并对其安全性和免疫保护效果进行测定.结果表明该疫苗安全可靠,对强毒攻击可产生较好的抵抗力,攻毒保护率达80%.疫苗在2℃～8℃保存12个月后仍能保持良好的免疫效果.     

山羊痘灭活疫苗研究

用对山羊痘病毒敏感的原代或次代牛睾丸细胞进行病毒增殖培养,从中筛选出一株进行培养,将获得的病毒液用二乙烯亚胺作为灭活剂制备病毒灭活疫苗接种山羊。分别在接种后7、14、21、28及72d采集血清用琼脂扩散试验进行免疫抗体检测,均检测到较强的沉淀抗体;在72d对山羊进行强毒攻击试验未出现山羊痘病理变化。实验证明疫苗安全性良好,制备的山羊痘灭活疫苗对山羊有较强免疫保护作用。     

猪丹毒丝菌灭活疫苗免疫佐剂的筛选

旨在筛选适宜于猪丹毒丝菌灭活疫苗的佐剂。以分离鉴定出的猪丹毒丝菌1a型HG-1株灭活菌体为抗原分别配制矿物油佐剂疫苗(简称矿物油疫苗)、氢氧化铝胶佐剂疫苗(简称铝胶疫苗)、ISA201双相油乳佐剂疫苗(简称ISA201疫苗)、GEL02水溶性聚合物佐剂疫苗(简称GEL疫苗)、IMS1313水溶性纳米佐剂疫苗(简称IMS1313疫苗)共5种佐剂的灭活疫苗。小鼠免疫保护试验结果表明,二免14 d后使用约为4 LD_(50)的HG-1株对小鼠进行腹腔攻毒,矿物油疫苗和GEL疫苗的保护率分别为100%(7/7)和71%(5/7),其他三种佐剂疫苗的保护率均为14%(1/7)。本研究进一步选择铝胶疫苗和GEL疫苗进行猪体对比试验;仔猪安全性试验结果表明,两种佐剂疫苗的副反应均较小;免疫保护试验结果表明,两次免疫后使用约为16 LD_(100)的HG-1株对免疫仔猪进行耳缘静脉攻毒,两种佐剂疫苗的保护率分别为60%(3/5)和100%(5/5)。本研究最终选择GEL佐剂作为开发猪丹毒灭活疫苗的最适佐剂。     

貂源伪狂犬病毒DL14/08株免疫原性及其保护效果的研究

为研制貂源伪狂犬病(PR)灭活疫苗,本实验室从疑似患PR的水貂脑组织中分离到一株PR病毒(PRV)DL14/08株(10~(7.10)TCID_(50)/m L),采用甲醛灭活以铝胶为佐剂制备了水貂源PR灭活疫苗。采用水貂和家兔对疫苗进行安全性检验和最小免疫剂量测定,结果显示水貂和家兔的最小免疫剂量均为5×10~(5.6)TCID_(50)/m L;采用研制的PR灭活疫苗和商品化猪用PR灭活疫苗免疫水貂,免疫21 d后平均中和抗体分别为1∶516和1∶348,用相当于100倍半数致死量毒力的分离株病毒攻毒,自制灭活疫苗组保护率为100%,商品化疫苗组保护率为80%。实验结果表明,制备的水貂源PR灭活疫苗抗体水平及攻毒后保护率明显高于商品化疫苗,能够有效保护强毒株对水貂的攻击,可以作为水貂伪PRV预防和控制的候选疫苗株。     

猪丹毒丝菌灭活疫苗免疫佐剂的筛选

旨在筛选适宜于猪丹毒丝菌灭活疫苗的佐剂。以分离鉴定出的猪丹毒丝菌1a型HG-1株灭活菌体为抗原分别配制矿物油佐剂疫苗（简称矿物油疫苗）、氢氧化铝胶佐剂疫苗（简称铝胶疫苗）、ISA201双相油乳佐剂疫苗（简称ISA201疫苗）、GEL02水溶性聚合物佐剂疫苗（简称GEL疫苗）、IMS1313水溶性纳米佐剂疫苗（简称IMS1313疫苗）共5种佐剂的灭活疫苗。小鼠免疫保护试验结果表明,二免14 d后使用约为4 LD₅₀的HG-1株对小鼠进行腹腔攻毒,矿物油疫苗和GEL疫苗的保护率分别为100%（7/7）和71%（5/7）,其他三种佐剂疫苗的保护率均为14%（1/7）。本研究进一步选择铝胶疫苗和GEL疫苗进行猪体对比试验;仔猪安全性试验结果表明,两种佐剂疫苗的副反应均较小;免疫保护试验结果表明,两次免疫后使用约为16 LD₁₀₀的HG-1株对免疫仔猪进行耳缘静脉攻毒,两种佐剂疫苗的保护率分别为60%（3/5）和100%（5/5）。本研究最终选择GEL佐剂作为开发猪丹毒灭活疫苗的最适佐剂。     

羊源致病性柠檬酸杆菌4种佐剂灭活疫苗的免疫效果比较

为筛选羊源弗氏柠檬酸杆菌灭活菌体疫苗的合适佐剂,本研究以灭活羊源弗氏柠檬酸杆菌LK-1株为抗原分别制备其矿物油疫苗、铝胶疫苗、GEL疫苗和ISA201疫苗。将4种不同佐剂疫苗分别背部皮下免疫小鼠(0.2 mL/只)和颈部肌肉免疫羊(2 mL/只)进行疫苗安全性试验。用4种不同佐剂疫苗分别背部皮下免疫小鼠(0.2 mL/只)两次后使用3 LD50的LK-1株进行腹腔攻击;颈部肌肉免疫羊(2 mL/只)两次后使用2个致死剂量的LK-1株进行耳静脉攻击,比较4种不同佐剂疫苗对小鼠和羊的保护率并对小鼠和羊免疫血清进行微量凝集抗体检测。疫苗安全性试验结果显示,矿物油疫苗导致接种动物出现严重的精神沉郁、食欲减退、以及接种部位发生肿胀坏死和疫苗残留等不良反应;其它3种佐剂疫苗不良反应较轻微。动物免疫试验结果显示,4种佐剂疫苗对小鼠的攻毒保护率分别为85%、50%、80%、100%,对羊的保护率分别为100%、75%、0、100%。4种佐剂疫苗综合比较,ISA201疫苗使接种动物产生最轻微的副反应,而且能产生较高的抗体水平和最强的保护力。本研究为进一步开发羊柠檬酸杆菌病的灭活疫苗奠定基础。     

猪链球菌三价灭活疫苗免疫佐剂的筛选

为筛选适用于新型猪源链球菌(SS)灭活疫苗的佐剂,本研究以SS1 Z1株、SS2 Z2株和SS7 Z7株为抗原,分别配制ISA201 VG双相油乳佐剂疫苗(简称ISA201疫苗)、GEL 02 PR水溶性聚合物佐剂疫苗(简称GEL疫苗)、IMS1313N VG水溶性纳米佐剂疫苗(简称IMS1313疫苗)、矿物油佐剂疫苗(简称油疫苗)、氢氧化铝胶佐剂疫苗(简称铝胶疫苗)共5种佐剂的三价灭活疫苗。豚鼠和仔猪安全性试验结果表明:油佐剂疫苗的副反应较大,其余疫苗的副反应较小。仔猪免疫保护试验结果显示:二免14 d后利用约为1 LD_(100)的SS2 Z2株对仔猪经耳缘静脉攻毒,ISA201疫苗、GEL疫苗对仔猪的保护率均为100%,油疫苗和铝胶疫苗保护率分别为60%、40%,IMS1313疫苗组的保护率为0;利用1 LD_(100) SS1 Z1株攻毒后,ISA201疫苗、GEL疫苗、油疫苗、铝胶疫苗和IMS1313疫苗对仔猪的保护率分别为100%、80%、80%、60%和0。综上结果表明,ISA201疫苗免疫仔猪能产生最高的抗体水平,同时提供了最高的保护效力,且副反应较小,确定ISA201为开发链球菌灭活苗的首选佐剂。本研究为SS三价灭活疫苗的研发奠定了基础。     

CpG寡核苷酸增强鸡新城疫疫苗免疫效力的研究

为了探讨CpG寡核苷酸（CpG oligonucleotide,CpG ODN）对鸡新城疫疫苗免疫效力的影响,将CpG2007与鸡淋巴细胞共孵育,测定淋巴细胞增殖率,结果发现CpG2007对鸡淋巴细胞具有显著的刺激活性。将CpG2007与不同浓度的新城疫抗原混合,制备灭活疫苗,免疫健康雏鸡。分别于免疫后不同时间采血,测定抗体效价和细胞因子表达量,并进行攻毒保护试验。结果发现,添加CpG ODN佐剂的试验组均比对应相同抗原剂量的免疫对照组的抗体水平高,产生抗体速度快;抗原剂量降低10倍的佐剂试验组与高抗原剂量免疫对照组抗体水平和攻毒保护率均相当,表明CpG ODN能显著增强新城疫疫苗的免疫效力,能促进机体产生更强烈的免疫应答,是有效的疫苗佐剂候选物质。     

鸽禽Ⅰ型副粘病毒油佐剂灭活苗对雏鸡免疫效果评价

用鸽Ａ／ＰＭＶ－１油佐剂灭活苗与ＮＤＶ油佐剂灭活苗分别免疫雏鸡,免疫后２１ｄ抗体水平达到峰值,免疫后４２ｄ用新城疫强毒对两种疫苗免疫鸡分别进行攻击,鸽Ａ／ＰＭＶ－１油佐剂灭活苗免疫组保护率为７３．３３％,ＮＤＶ油佐剂灭活苗免疫组保护率为９９．６７％。     

Efficacy of an inactivated recombinant vaccine encoding a fimbrial protein of Pasteurella multocida B:2 against hemorrhagic septicemia in goats

This study was carried out to determine the antibody responses and protective capacity of an inactivated recombinant vaccine expressing the fimbrial protein of Pasteurella multocida B:2 following intranasal vaccination against hemorrhagic septicemia in goats. Goats were vaccinated intranasal with 10⁶ CFU/mL of the recombinant vaccine (vaccinated group) and 10⁶ CFU/mL of pET32/LIC vector without fimbrial protein (control group). All three groups were kept separated before all goats in the three groups were challenged with 10⁹ CFU/mL of live pathogenic P. multocida B:2. During the course of study, both serum and lung lavage fluid were collected to evaluate the antibody levels via enzyme-linked immunosorbent assay. It was found that goats immunized with the inactivated recombinant vaccine developed a strong and significantly (p < 0.05) higher specific IgA and IgG responses in both serum and lung lavage fluid samples compared to the control and unvaccinated groups. Following intratracheal challenge, the rate of isolation was 17% for the vaccinated group, 67% for the control group and 100% for the unvaccinated group. However, none of the goat from the vaccinated group had P. multocida B:2 in the liver, tonsil and heart. Therefore, the study revealed that an inactivated recombinant vaccine significantly provides significant protection against high dose challenge and enhances the stimulation of the local and systemic immunities.     

Mucosal vaccination with formalin-inactivated avian metapneumovirus subtype C does not protect turkeys following intranasal challenge

Studies were performed to determine if mucosal vaccination with inactivated avian metapneumovirus (aMPV) subtype C protected turkey poults from clinical disease and virus replication following mucosal challenge. Decreases in clinical disease were not observed in vaccinated groups, and the vaccine failed to inhibit virus replication in the tracheas of 96% of vaccinated birds. Histopathologically, enhancement of pulmonary lesions following virus challenge was associated with birds receiving the inactivated aMPV vaccine compared to unvaccinated birds. As determined by an enzyme-linked immunosorbent assay (ELISA), all virus-challenged groups increased serum immunoglobulin (Ig) G and IgA antibody production against the virus following challenge; however, the unvaccinated aMPV-challenged group displayed the highest increases in virus-neutralizing antibody. On the basis of these results it is concluded that intranasal vaccination with inactivated aMPV does not induce protective immunity, reduce virus shedding, or result in decreased histopathologic lesions.     

鸡新城疫病毒分离株与La Sota株灭活疫苗效力比较试验

用NDV分离株及La Sota株为抗源液，经福尔马林灭活后，与油佐剂混合，分别制成分离株灭活苗、La Sota株灭活苗及分离株与La Sota株二价灭活苗。将这三种灭活疫苗分别免疫SPF鸡后，均获得100%抵抗NDV分离株及F48株强毒攻击的保护力；而用这3种灭活苗与La Sota活苗单独或联合使用，免疫带有ND母源抗体的普通鸡后，3种灭活苗的免疫效力不同，分离株灭活苗与价灭活苗对NDV分离株攻击的免疫保护效力明显优于La Sota灭活苗；灭活苗与活苗同时使用，其免疫效力明显优于单独使用灭活苗或活苗。     

鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活 疫苗(La Sota株+M41株+SS/94株)免疫程序研究

设计不同的免疫程序,用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)免疫黄羽肉鸡,通过对ND、IB、H9抗体滴度监测,探讨ND、IB、H9抗体消长规律及鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)的免疫程序。试验结果表明,用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)免疫黄羽肉鸡后,其诱导产生的ND、IB、H9抗体的消长规律基本同步;仅于10日龄免疫一次三联灭活苗,其抗体水平较低,于20、40日龄二免、三免或10日龄先用鸡新城疫病毒(La Sota株)、禽流感病毒(H9亚型,SS/94株)二联灭活疫苗作基础免疫,20或40日龄再用三联灭活苗作加强免疫,则上述3种抗体均快速上升,且维持时间长。根据试验结果,建议按照正常免疫程序作基础免疫的健康肉鸡,饲养期较短的可于20日龄左右用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)作加强免疫,0.3 mL/只;饲养期较长的则于40日龄左右用鸡新城疫、传染性支气管炎、禽流感(H9亚型)三联灭活疫苗(La Sota株+M41株+SS/94株)作加强免疫,0.5 mL/只。     

鸽新城疫油乳剂灭活苗的研制

本文报导了一种源自病鸽的鸽新城疫病毒ＱＬ株制备的灭活油乳剂疫苗。以此种疫苗接种后,接种鸽均正常无任何病症,接种疫苗获得保护能抵抗鸽新城疫病毒株攻击的鸽子数多于接种ＬａＳｏｔａ油乳剂疫苗的鸽子。已在野外以此种疫苗接种了大量的鸽子,效果满意     

Acute arthritis in caprine arthritis-encephalitis virus challenge exposure of vaccinated or persistently infected goats

Goats vaccinated with inactivated caprine arthritis-encephalitis virus (CAEV) developed more severe arthritis after infectious CAEV challenge exposure than did goats vaccinated with tissue culture medium. Arthritis also developed more rapidly in the group vaccinated with inactivated virus. In another experiment, goats with persistent CAEV infection developed acute arthritis after at least 2 injections of infectious CAEV at monthly intervals. In this experiment, the control group consisted of goats with persistent CAEV that were given tissue culture medium. Seemingly, the immune response to CAEV is an important cause of the CAEV-induced arthritis.     

Use of the caprinised strain of rinderpest virus for potency testing an attenuated cell culture rinderpest vaccine

The caprinised strain of rinderpest virus was inoculated into goats to produce a challenge stock. These goats were kept with control animals (goats, sheep, calves). In this trial the caprinised strain was shown to have a mild pathogenicity for goats and it spread to one of two contact goats but not from goats to other species. The caprinised strain was then tested on cattle where a febrile reaction was observed. The caprinised strain also did not spread between cattle. The cattle vaccinated with a freeze-dried vaccine produced from the attenuated Kabete RBKO strain on bovine kidney cells were then challenged with the caprinised strain with good results.     

Enhancement of the immune response and virological protection of calves against bovine herpesvirus type 1 with an inactivated gE-deleted vaccine

Four immunisation protocols based on inactivated and attenuated commercially available marker vaccines for bovine herpesvirus type 1 (BHV-1) were compared. The first group of calves were vaccinated with an attenuated vaccine administered intranasally and an inactivated vaccine injected subcutaneously, four weeks apart; the second group were vaccinated twice with the attenuated vaccine, first intranasally and then intramuscularly; the third group were vaccinated twice subcutaneously with the inactivated vaccine; and the fourth group were vaccinated twice intramuscularly with the attenuated vaccine. A control group of calves were not vaccinated. The cellular and humoral immune responses were highest in the two groups which received at least one injection of the inactivated vaccine. Virological protection was observed in all the vaccinated groups after a challenge infection and reactivation by treatment with dexamethasone, but the calves which received one dose of the inactivated vaccine as a booster or two doses of the inactivated vaccine excreted significantly less of the challenge virus than the calves which were vaccinated only with the attenuated preparation.