“兔瘟”疫苗中不同佐剂对免疫效果的影响

利用“兔瘟”病毒枣(?)毒株(RVZ85)制备的不含佐剂疫苗,甘油佐剂疫苗、氢氧化铝佐剂疫苗和矿物油佐剂疫苗免疫接种易感家兔,定期测 HI 抗体效价并进行人工攻毒试验.结果证明,不含佐剂的疫苗产生的保护力最早,免疫后三天即4/4保护.用四种疫苗免疫接种后一年内,对攻毒的保护率均为100%.四种疫苗中,以矿物油佐剂疫苗免疫兔的HI抗体效价最高,氢氧化铝佐剂疫苗次之,甘油佐剂疫苗及不含佐剂疫苗的HI效价较低。氢氧化铝佐剂疫苗在室温保存8.5个月,免疫兔4/4保护;室温保存14个月.免疫兔2/3保护:4℃保存14个月,免疫兔4/4保护.     

新生仔猪注射猪瘟兔化弱毒疫苗的效果观察

对新生仔猪在哺乳前及吸吮初乳后3、6、9、12和15小时分别接种猪瘟兔化弱毒疫苗,待60日龄时测定其抗猪瘟抗体效价并进行攻毒,观察其免疫效果。结果,哺乳前及吃初乳后3、6和9小时免疫组抗体效价分别是1:128、1:128、1:64和1:32以上.免疫保护率分别为100%、100%、100%和75%。12、15小时免疫组和对照组的抗体效价分别是1:8、1:4和1:8以下,免疫保护率均为零。     

兔源抗IBDV独特型抗体疫苗的制备

用纯化的抗IBDV抗体作为抗原免疫家兔,将分离制得的致价（1：16）较高的兔源抗IBD独特型抗体分别与福氏完全和福氏不完全佐剂按1：1比例乳化制备成抗IBDV独特型抗体疫苗,免疫接种普通迪卡公雏鸡和SPF鸡,然后用IBDV强毒株经点眼和滴鼻方式攻毒,试验组的保护率分别为98／98、100／100、49／49,大群免疫接种应答为100％,具有良好的免疫保护效果。由此也证实了抗独特型抗体疫苗的制备潜在的研究和应用价值。     

鸡沙门氏菌外膜蛋白亚单位疫苗的研究

用十二烷酰肌氨酸法提取鸡沙门氏菌外膜蛋白（OMP）作为抗原，制备免疫激发复合物型（Immunostimulating Complex,ISCOM）亚单位疫苗。ISCOM型OMP亚单位疫苗在电镜下可笼型颗粒结构，直径为60mm-120nm。无菌性检测结果为无菌，具有可靠的安全性。对3周龄雏鸡肌注接种，6周龄攻毒，接种80ugOMP／只ISCOM型亚单位疫苗的雏鸡以鸡白痢沙门氏菌或鼠伤寒沙门氏菌的攻击具有显著的免疫保护力。     

猪伪狂犬病不同佐剂灭活疫苗对兔免疫原性初探

采用分离鉴定的猪伪狂犬病毒（HB—J株）以4种佐剂研制成4批灭活疫苗,以研究其对兔的免疫原性。疫苗分别免疫PRV抗体阴性兔后,用乳胶凝集试验法（LAT）和中和试验法（SNT）对试验兔进行血清抗体检测;于免疫后21、28d对试验兔分别进行攻毒,观察兔保护情况。试验结果表明,兔对不同佐剂的猪伪狂犬病疫苗均产生良好的免疫应答反应,且当兔免疫后血清凝集价≥1：32或血清抗体中和指数≥1479或中和价≥1：16时,可以抵抗10LD50剂量强毒的攻击;当兔免疫后血清凝集价≥1：64或血清抗体中和指数≥2187或中和价≥1：32时,可以抵抗100LD50剂量强毒的攻击;4种佐剂灭活疫苗均具有良好的免疫效果。     

罗非鱼无乳链球菌疫苗研制及检测IgM抗体ELISA方法的建立

为研究新型罗非鱼链球菌疫苗,并且建立检测罗非鱼无乳链球菌IgM抗体水平的ELISA方法,试验首先纯化了罗非鱼血清中的IgM,制备抗IgM的兔高免血清,优化反应条件,比较不同浓度浸泡型疫苗和注射型疫苗的相对免疫保护率,并利用建立的ELISA方法监测罗非鱼免疫后2周抗体水平以及攻毒后4个月内的免疫鱼血清抗体水平变化。结果表明:高浓度浸泡型疫苗能达到88. 65%的相对保护率; ELISA方法特异性较高,重复试验中变异系数最大值为8. 33%,灵敏度为1∶400。通过对免疫鱼血清抗体的跟踪监测,表明血清抗体在免疫后2周明显上升,且产生的免疫保护力可长达4个月之久。说明建立的ELISA方法可以检测罗非鱼IgM抗体,制备的疫苗通过浸泡的方式免疫,可使罗非鱼获得长达4个月的保护力。     

兔病毒性出血症（RHD）母源抗体动态与抗病毒感染关系

为了阐明母源抗体动态与抗病毒感染关系、母源抗体对疫苗免疫干扰程度，从而提出适宜的免疫程序，进而为易感动物筛选，疫苗免疫后抗体水平的跟踪调查等提供科学依据，进行了本项研究。 1试验材料和方法 1. 1被检血清样品 RHD常规免疫母兔怀孕到产期剖腹取胎兔，产后 24～ 48h未经吃初乳仔兔，已吃初乳仔兔，产后第 6、 15、 30、 45、 60、 90、 120日龄兔和产仔母兔，经心脏采血分离血清。 1. 2 RHD种毒 种毒系本研究组分离保存的兔病毒性出血症病毒 (RHDV)湖 84－ 12毒株。 1. 3凝集抗原 取人工感染死亡兔肝组织，以 PBS作 1∶ 10…     

兔出血症不同佐剂灭活疫苗的制备及免疫效果比较

本试验制备了3种兔出血症灭活苗,经对试验兔进行免疫对比试验,以1mL为免疫剂量,7d后检测,发现3种疫苗均可产生抗体。组织灭活苗、蜂胶佐剂苗、铝胶佐剂苗7d的抗体滴度分别为27.1、27.2、26.5;14d的抗体滴度分别为27.9、29.0、28.0;45d的抗体滴度分别为26.8、28.5、27.5;85d的抗体滴度分别为25.5、27.2、26.2;145d的抗体滴度普遍下降。在免疫攻毒试验中,第7、15、50、90d攻毒时,3种疫苗的保护率均为100%,对照组全部死亡;150d攻毒时,组织灭活苗、蜂胶佐剂苗、铝胶佐剂苗的保护率分别为25%、75%、50%。以上结果表明,3种疫苗都有很好的免疫效果,其中以蜂胶佐剂苗的抗体滴度更高,免疫保护期更长。     

自制兔瘟疫苗效力试验前后体温变化的初步观察

实验组兔用85—2批疫苗攻毒前后的体温变化不大,而未经免疫的对照组兔攻毒前后体温变化显著.免疫的长毛兔经120天攻毒后,均能耐受1—5毫升1:2同源肝毒的攻击,保护率达100%,而未经免疫的对照组攻毒后34—36小时全部死亡.     

新剂型兔瘟疫苗的研究

经血凝抑制和攻毒试验结果表明，在含有相同抗原量制备的疫苗，兔瘟油乳剂灭活疫苗比兔瘟组织灭活苗免疫后血清ＨＩ抗体效价高３．２５￣６．０ｌｏｇ２，即使油乳剂灭活疫苗仅为组织灭活苗四分之一的抗原量，免疫后血清ＨＩ抗体效价还高１ｌｏｇ２。油乳剂灭活苗免疫兔第５０天其血清ＨＩ抗体未见下降，而组织灭活苗免疫后第２６天已开始下降。虽然接种二种剂型的疫苗攻毒后均具有１００％的保护率，但血清ＨＩ抗体效价消长情况证明     

Vaccination of ferrets (Putorius furo) against rabies using a tissue vaccine

The use of live tissue rabies vaccine from Vnukovo-32 strain produced by the Bioveta National Corporation at Ivanovice na Hané was tested on 15 ferrets (Putorius furo L. 1758). The 21st day after subcutaneous application of 3 ml of vaccine, the average titre of virus-neutralizing antibodies was 1 : 71.4, and after subcutaneous and intramuscular application of 1 ml of vaccine the average titre was 1:20.6 and 1 : 15.6, respectively. The challenge test was performed 52 weeks after vaccination: a dose of 10(5) MICLD50 of the street rabies virus was applied i. m. to the neck muscle. All the vaccinated animals survived. As indicated by the results of the trials, a good tolerance and full immunity of vaccinates to the infection with the street rabies virus was maintained for one year. 1 ml of vaccine diluted according to instructions and applied s. c. or i. m. was found to be sufficient.     

Testing the effectiveness of foot-and-mouth disease vaccines: the relationship between test infection results and corresponding neutralization titers of vaccinated cattle

In the course of vaccine controls, the potency of 25 foot-and-mouth disease (FMD) vaccines was tested quantitatively in parallel in cattle using the intradermal infection and the determination of the SN titres. More than 95% of the vaccinated cattle with SN titres of greater than 1:20 were protected from generalized FMD, regardless of the virus type tested. 61.5% of the vaccinated cattle with SN titres less than or equal to 1:20 were not protected and developed generalized FMD. Comparison of the PD50 values calculated from the results of the intradermal infection and the corresponding SN titres (minimum protection titre greater than 1:20) showed that the results were in complete agreement in 56% of the tested vaccines. In a further 32% of vaccines, the PD50 calculated from the SN titre was slightly below that for the intradermal infection, in the remaining 12% it was somewhat above. The possibility of using the minimum titre determination for testing a vaccine and the significance of this titre as an expression of protection by vaccination are discussed.     

Testing for immunogenic and protective properties of vaccine against tissue-inactivated rabies

The immunogenic and protective potency was tested of vaccine against inactivated tissue rabies developed from the Vnukovo-32 strain, and produced in the Bioveta state corporation at Ivanovice in Haná. In 21 days after an i. m. application of 3 cm3 of vaccine, the average titre of virus-neutralizing antibodies was found to be 1:47. In this period the animals were revaccinated in the same way. The average titre of virus-neutralizing antibodies was 1:99 three months after revaccination, 1:57 in six months and 1:24 in nine months. In this period a challenge test was performed in a dog using the dose of 10(5) MICLD50 of street rabies virus. This dose was implanted i. m. into masticatory muscles. Another dog was infected experimentally 18 months after immunization. The experiment has proved the good immunogenic potency of vaccine against inactivated tissue rabies and its ability to induce the protection of vaccinated dogs from the strees infection with street rabies virus. The control rabies vaccine of foreign make RABISIN was tested in a similar way.     

An investigation into the viability of broth cultures of the T1 strain of Mycoplasma mycoides sub-species mycoides.

The T1 broth vaccine against contagious bovine pleuropneumonia can only be stored for four weeks at +4 degrees C because after that time the titre of Mycoplasma mycoides sub-species mycoides falls below the minimal vaccinating dose. An investigation into the death of these organisms during the stationary growth phase was made. The culture medium in which the vaccine was prepared was found to contain adequate nutrients. Control of the pH by the addition of KOH was shown to preserve the viability of the organism: there was no drop in titre during 336 h (14 days) at 37 degrees C. Use of this technique may prove valuable as a means of preserving the viability of T1 broth vaccine.     

A live attenuated glycoprotein E negative bovine herpesvirus 1 vaccine induces a partial cross-protection against caprine herpesvirus 1 infection in goats

Taking into account the close antigenic relationship between bovine herpesvirus 1 (BoHV-1) and caprine herpesvirus 1 (CpHV-1), a live attenuated glycoprotein E (gE) negative BoHV-1 vaccine was assessed in goats with the aim to protect against CpHV-1 infection. Vaccine safety was evaluated by intranasal inoculation of two groups of goats with either a gE-negative BoHV-1 vaccine or a virulent BoHV-1. The length of viral excretion and the peak viral titre were reduced with the gE-negative vaccine. To assess the efficacy, two goats were inoculated intranasally twice 2 weeks apart with a gE-negative BoHV-1 vaccine. Four weeks later, immunised and control goats were challenged with CpHV-1. A 2 log(10) reduction in the peak viral titre was observed and the challenge virus excretion lasted 2 days more in immunised than in control goats. These data indicate the safety and the partial efficacy of a live attenuated gE-negative BoHV-1 vaccine intranasally administrated in goats.     

Freeze-dried vaccine against Rinderpest: Stability and activity study

A freeze-dried vaccine against Rinderpest was prepared from modified virus multiplied in calf kidney cell cutlure. Characteristics of the vaccine are as follows:

• —high titre after freeze-drying (10⁴ CCID₅₀/dose),

• —well-adapted freeze-drying stabiliser which ensures maintenance of the infective titre of the vaccinal virus, even under severe conditions (3.5 days at +45°C),

• —use of an appropriate solvent: magnesium sulphate molar solution or more simply physiological saline (for stability after reconstitution even at high temperatures—up to 4 h at +45°C).

The activity of the vaccine, tested in cattle by antibody titration and resistance to specific challenge perfectly satisfies requirements set by the WHO and OIE.     

鸡传染性鼻炎灭活油乳剂苗的研制

采用抗原性较好的Ａ型和Ｃ型菌株,以矿物油为佐剂制备成Ａ、Ｃ型二价油乳剂灭活菌苗,肌内接种健康鸡,玻片凝集试验表明,免疫后第七天产生凝集抗体,效价达到１：４以上,第十六天效价可达１;８,第二十四天效价高达１：１６,攻毒试验的结果表明,抗体效价达到１：１６时可以抵抗强毒的攻击,田间试验的结果表明,该苗的免疫原性好,安全。     

Duration of protective immunity and antibody responses in cattle immunised against alcelaphine herpesvirus-1-induced malignant catarrhal fever

ABSTRACT: Protection of cattle from alcelaphine herpesvirus-1 (AlHV-1)-induced malignant catarrhal fever (MCF) has been described previously, using an attenuated virus vaccine in an unlicensed adjuvant. The vaccine was hypothesised to induce a protective barrier of virus-neutralising antibody in the oro-nasal region, supported by the observation of high titre neutralising antibodies in nasal secretions of protected animals. Here we describe further analysis of this vaccine strategy, studying the effectiveness of the vaccine formulated with a licensed adjuvant; the duration of immunity induced; and the virus-specific antibody responses in plasma and nasal secretions. The results presented here show that the attenuated AlHV-1 vaccine in a licensed adjuvant protected cattle from fatal intranasal challenge with pathogenic AlHV-1 at three or six months. In addition, animals protected from MCF had significantly higher initial anti-viral antibody titres than animals that succumbed to disease; and these antibody titres remained relatively stable after challenge, while titres in vaccinated animals with MCF increased significantly prior to the onset of clinical disease. These data support the view that a mucosal barrier of neutralising antibody blocks infection of vaccinated animals and suggests that the magnitude of the initial response may correlate with long-term protection. Interestingly, the high titre virus-neutralising antibody responses seen in animals that succumbed to MCF after vaccination were not protective.     

Antibody response to an anti-rabies vaccine in a dog population under field conditions in Bolivia

Rabies remains an important public health issue in Bolivia, South America. Public concern and fears are most focussed on dogs as the source of rabies. The objective of the present study was to assess immunity of an inactivated suckling mouse brain vaccine against canine rabies used for the official vaccination campaigns under field conditions in an endemic area of rabies in Bolivia. A total of 236 vaccinated and 44 unvaccinated dogs in Santa Cruz de la Sierra, selected using stratified random sampling, were investigated in order to obtain owned dog characteristics and antibody titres against rabies in April 2007. The proportion of vaccinated dogs with an antibody titre exceeded the protection threshold value of 0.5 EU/ml was 58% [95% confidence intervals (CI): 52-65], indicating that vaccination is likely to elicit an antibody response (odds ratio 6.3, 95% CI: 1.2-11.5). The range of geometric mean of antibody titre for vaccinated dogs (0.89 EU/ml; 95% CI: 0.75-1.04) was considered to meet the minimal acceptable level indicating an adequate immune response to the vaccine. However, the titre level was not satisfactory in comparison with the results from other field investigations with inactivated tissue culture vaccines. It is recommended for public health authorities to (1) consider modernizing their vaccine manufacturing method because the level of immunity induced by the current vaccine is comparably low, (2) conduct frequent vaccination campaigns to maintain high levels of vaccination coverage, and (3) actively manage the domestic dog population in the study area, which is largely responsible for rabies maintenance.     

An evaluation of the role of antibodies to Actinobacillus pleuropneumoniae serovar 1 and 15 in the protection provided by sub-unit and live streptomycin-dependent pleuropneumonia vaccines

OBJECTIVE: To evaluate the serological response of pigs receiving either the Porcilis APP vaccine or a modified live vaccine based on a streptomycin-dependent (SD) strain of Actinobacillus pleuropneumoniae, and then challenged with an Australian isolate of A. pleuropneumoniae of either serovar 1 or 15 as a means of understanding the protection provided by both vaccines against serovar 1 but not against serovar 15. DESIGN: The serological tests evaluated were serovar-specific polysaccharide ELISA tests (for serovar 1 and 15), ELISA tests for antibodies to three A. pleuropneumoniae toxins (ApxI, ApxII and ApxIII) as well as to a 42 kDa outer membrane protein (OMP), a haemolysin neutralisation (HN) assay and immunoblotting. The tests were used to detect antibodies in vaccinated pigs that had been shown to be protected against serovar 1 but not serovar 15. RESULTS: In the polysaccharide antigen ELISA assays, both vaccines resulted in a significant rise in the titre in the serovar 1 ELISA but not the serovar 15 ELISA. The Porcilis APP vaccinated pigs showed a significant response in the ApxI, ApxIII and 42 kDa OMP ELISA. In the ApxII ELISA, all pigs tested (the Porcilis APP vaccinates and the controls) were positive on entry to the trial. In the HN assay, the Porcilis APP vaccinated pigs showed a significant response after one dose while the SD vaccinated pigs required two doses of vaccine before a marked rise in titre was induced. Immunoblotting revealed that neither vaccine generated antibodies that recognised the ApxIII produced by serovar 15. CONCLUSIONS: The failure of these vaccines to provide protection against serovar 15 may be due to novel virulence factors possessed by serovar 15, significant differences between the ApxIII toxin of serovar 15 and those present in the Porcilis APP vaccine or failure by both vaccines to induce antibodies to the serovar 15 specific polysaccharide.