Duration of immunity with Mycoplasma synoviae: comparison of the live attenuated vaccine MS-H (Vaxsafe MS) with its wild-type parent strain, 86079/7NS

The duration of protective immunity elicited by the MS-H vaccine was evaluated by experimental challenge of chickens at 15 and 40 wk after eyedrop vaccination. Immunity induced by the parent strain of the vaccine, 86079/7NS, was also investigated for comparison. A serological response to Mycoplasma synoviae was detected in 89% to 100% of MS-H vaccinates and 86079/7NS inoculates at 15, 27, 30, 35, and 40 wk after inoculation. A significantly lower incidence of air-sac lesions and lower air-sac lesion severity were observed in both the MS-H vaccinated and the 86079/7NS inoculated groups, as compared to the unvaccinated controls, after both challenge points. Tracheal mucosal thicknesses in MS-H vaccinates was significantly lower in the upper, lower, and total trachea at 40 wk after vaccination, as compared to the controls. It was demonstrated in this experiment that protective immunity, as determined by protection against experimental challenge, was maintained to at least 40 wk after vaccination.     

Determination of the effective dose of the live Mycoplasma synoviae vaccine, Vaxsafe MS (strain MS-H) by protection against experimental challenge

The minimum effective dose of the Mycoplasma synoviae-H (MS-H) vaccine was determined through protection against experimental challenge. Chickens were vaccinated by eyedrop with the following doses of a vaccine: 1.2 x 10(5), 2.4 x 10(5) 4.8 x 10(5), 9.6 x 10(5), 1.92 X 10(6), and 3.84 X 10(6) color change units (CCU), then challenged 6 wk after vaccination. Rapid serum agglutination results indicated that 100% of birds receiving an MS-H dose of > or = 4.8 x 10(5) CCU had antibodies to MS and enzyme-linked immunosorbent assay results showed that 60% of birds receiving a dose of 4.8 x 10(5) or 9.6 x 10(5) CCU and 100% of birds receiving a dose of 1.92 x 10(6) or 3.84 x 10(6) had antibodies to MS. At postmortem after challenge, the following parameters were significantly lower in birds vaccinated with an MS-H dose of > or = 4.8 x 10(5) CCU: air sac (AS) lesion severity; incidence of AS lesions; mucosal thicknesses in the upper trachea, middle trachea, and lower trachea (LT); and MS colonization of the LT and AS. It was concluded that an MS-H dose of 4.8 x 10(5) CCU was sufficient to elicit an antibody response in birds, prevent MS colonization in the LT and AS, and protect against AS lesions caused by an experimental MS and infectious bronchitis virus challenge.     

Evaluation of the non-temperature-sensitive field clonal isolates of the Mycoplasma synoviae vaccine strain MS-H

The live attenuated temperature-sensitive (ts+) Mycoplasma synovia (MS) strain, MS-H, is used as a vaccine in a number of countries to control virulent MS infection in commercial chicken flocks. Nine out of 50 isolates made from flocks vaccinated with MS-H were found to have lost the ts+ phenotype of the original vaccine strain. In order to examine the influence of the ts- phenotype on virulence of the isolates, four of the ts- isolates, the MS-H vaccine, and the vaccine parent strain 86079/7NS were administered by aerosol in conjunction with infectious bronchitis virus to 3-wk-old specific-pathogen-free chickens. The four ts- clones induced only minimal air sac lesions that were not different in severity from those caused by MS-H vaccine; however, the vaccine parent strain 86079/7NS caused air sac lesions that were significantly greater than those of MS-H and all ts- clones. The vaccine parent strain 86079/7NS and two of the ts- clones were recovered from the air sacs of the respectively infected chickens whereas the MS-H vaccine and two other ts- clones were not. Three of the ts- isolates caused increased tracheal mucosal thicknesses that were significantly greater than those from birds inoculated with MS-H, and one caused increased tracheal mucosal thicknesses that were significantly less than those from birds inoculated with 86079/7NS. In conclusion, unlike the MS-H vaccine, the MS-H ts- clones were associated with minor changes in tracheal mucosa; however, unlike the vaccine parent strain, they did not induce lesions in the air sacs. These results suggest that factors other than ts+ phenotype are involved in the attenuation of the MS-H vaccine.     

Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene

1. The single-copy domain of the N-terminal region of the vlhA gene of Mycoplasma synoviae was sequenced, analysed and verified and used to type Iranian field isolates of M. synoviae and the MS-H live vaccine strain. In addition, a restriction fragment length polymorphism (RFLP) method was developed to differentiate between field isolates of Iranian and MS-H vaccine strains.

2. All sequences were analysed and aligned; the percentage similarity of the DNA was calculated and dendrograms were constructed. Based on single nucleotide polymorphism (SNP) that existed in all field isolates in Iran, the PCR-RFLP method allowed the differentiation of all M. synoviae field isolates from the vaccine strain.

3. Using phylogenetic analysis, the isolates were assigned to 8 unique genotypes and, within each group, DNA had a high level of similarity.

4. DNA sequence analysis and PCR-RFLP of the amplicon based on percent similarity and evolutionary relationship appeared to be useful tools for strain differentiation whether M. synoviae clinical isolates from infected chickens were derived from the vaccine strain or wild-type strains.

5. This study confirms the potential value of strain typing for epidemiological purposes and suggests that phylogenetic studies are essential to understand the true relationships between strains.     

Safety and efficacy of the Mycoplasma synoviae MS-H vaccine in turkeys

The live, attenuated, temperature-sensitive Mycoplasma synoviae (MS) vaccine strain MS-H is used to control virulent MS infection in commercial chicken flocks. However, the safety of this vaccine and its potential to prevent disease in turkeys have not been investigated. In this study, MS-H was shown to colonize the upper respiratory system and to induce an antibody response in turkeys but, even at the maximum release dose, was not found to cause air sac, joint, or tracheal lesions typical of wild-type MS infection. Histopathologic examinations of the vaccinated turkeys after exposure to a virulent MS challenge revealed that administration of the vaccine by aerosol, but not eye drop, at the dose recommended for chickens protected the birds against microscopic lesions and colonization of the virulent MS in trachea. It is concluded that MS-H vaccine is safe for use in turkeys and, when used as aerosol at the dose recommended for commercial chickens, can protect turkeys against tracheal lesions caused by a wild-type MS strain.     

Genotyping of Japanese field isolates of Mycoplasma synoviae and rapid molecular differentiation from the MS-H vaccine strain

Mycoplasma synoviae is an important causative agent of avian mycoplasmosis. In the present study the conserved domain of the variable lipoprotein and hemagglutinin (vlhA) gene of M. synoviae was sequenced and analyzed for 19 field strains of M. synoviae isolated from chickens across Japan. This analysis revealed that there were at least nine genotypes of M. synoviae present in Japan. Furthermore, we found a single nucleotide polymorphism (SNP) within this region in all the Japanese isolates, and based on this finding, we established a PCR method with cycling probe technology to differentiate between these field isolates and the live M. synoviae vaccine strain Mycoplasma synoviae-H (MS-H).     

A modified live PRRSV vaccine and the pathogenic parent strain induce regulatory T cells in pigs naturally infected with Mycoplasma hyopneumoniae

Rhodococcus equi is an important respiratory pathogen of young foals for which a vaccine has long been sought. Two major impediments to effective vaccination are the functionally immature type I immune responses of neonatal foals and early exposure to the bacterium via the environment. Despite these obstacles, it appears that under specific circumstances foals can develop a protective immune response. In this study we investigated the protective mechanisms behind oral inoculation of foals with virulent R. equi bacteria. Two foals receiving an oral inoculum demonstrated accelerated development of R. equi specific cytotoxic T lymphocytes (CTL) as evidenced by significant lysis of R. equi infected, ELA-A mismatched cells at 3 weeks of age. As in a previous study, CTL were not detected until 5-6 weeks of age in two control foals. At each time point the ability of foal peripheral blood mononuclear cells (PBMC) to produce IFN-γ following stimulation with live R. equi or extracted cell wall lipids was similar to that of an adult horse control and between foals, regardless of treatment. These results provide a potential mechanism of protection which has previously been shown to occur following oral inoculation, and suggest that the early detection of CTL may be a useful marker for induction of protective immunity.     

Protective efficacy of a live attenuated Mycoplasma hyopneumoniae vaccine with an ISCOM-matrix adjuvant in pigs

An attenuated Mycoplasma hyopneumoniae vaccine that requires intrathoracic administration is commercially available for use against mycoplasmal pneumonia in China. Given the limitations of such a route of administration, this study was undertaken to assess the capacity of an ISCOM-matrix adjuvant to enhance immunogenicity following intramuscular use. Immune responses in pigs following vaccination and subsequent intra-tracheal bacterial inoculation were examined using lymphocyte proliferation, serology and mucosal IgA in both nasal and saliva swabs.Vaccination induced clear lymphocyte proliferation, but only slight serum antibody responses although these were significantly increased following experimental infection. Mucosal IgA was not detected in either nasal or salivary secretions. Following bacterial challenge, animals vaccinated with the adjuvant-containing live vaccine exhibited less severe pulmonary lesions (median score 3.67) than unvaccinated pigs (median score 13.58). The degree of ciliary loss on the respiratory tract surface was reduced in vaccinated pigs compared with experimentally infected controls. The findings indicated that the adjuvant vaccine administered IM provided protection against experimentally induced mycoplasmal pneumonia and could have commercial potential.     

表达IBDV VP2蛋白重组减毒沙门菌活载体疫苗株的构建及免疫原性分析

通过PCR克隆出IBDV VP2基因,将其插入到表达载体pYA3341中,构建重组质粒pYA3341-VP2。将重组质粒电转入鼠伤寒沙门菌疫苗株X4550（缺失Asd、Cya、Crp基因）,获得重组疫苗菌株X4550（pYA3341-VP2）。进行重组菌VP2蛋白表达的鉴定;测定重组菌的稳定性、生长曲线、安全性以及小鼠免疫试验。结果表明,酶切鉴定证实重组质粒构建成功;SDS-PAGE和Western blot证实重组菌表达的VP2蛋白能与鸡抗IBDV阳性血清特异性结合;重组菌株在体外营养选择压力下,可稳定地携带重组质粒传代繁殖,在体内可稳定地定居于肠系膜淋巴结和脾脏;小鼠口服试验证实重组菌无毒性作用;口服重组菌免疫小鼠,ELISA检测产生了抗IBDV抗体;中和试验表明产生的抗体具有中和活性。本试验成功构建了能稳定表达IBDV VP2蛋白的口服减毒鼠伤寒沙门菌疫苗株X4550（pYA3341-VP2）,为研究IBD口服基因工程疫苗奠定了基础。     

Molecular differentiation between NS1 gene of a field strain Bluetongue virus serotype 2 (BTV-2) and NS1 gene of an attenuated BTV-2 vaccine

At the end of September 2000, clinical symptoms of Bluetongue appeared in sheep flocks of the Balearic Islands (Spain). The presence of the BTV serotype 2 in tissue and blood samples of affected animals was confirmed by laboratory techniques. A systematic vaccination were carried out in affected areas using a live monovalent serotype 2 vaccine available from Onderstepoort laboratory (South Africa). In order to perform epidemiological studies, a new method to differentiate between the NS1 genes of BTV-2 affecting the Balearic islands and that of the Onderstepoort commercial live virus vaccine (monovalent, serotype 2) has been developed. This procedure is based on the use of an RT-PCR, followed by restriction endonuclease analysis. Epidemiological data of a study carried out in the period January-October 2001 using this procedure are included.     

鸡马立克氏病CVI 988/Rispens毒株生物学特性的研究

对荷兰农业渔业部提供的初代次 CVI988/ Rispens种毒 ,繁殖到 37代 ,用间接荧光抗体试验测定属血清 型 ,经接种雏鸡进行 6代毒力返强试验观察到 70日龄和对 1日龄雏鸡接种 4 5 0 0 0个蚀斑单位 ,观察到 12 0日龄均为安全。对 1日龄雏鸡接种免疫剂量的疫苗 ,于 4日龄产生病毒血症 ,7～ 14日龄达到高峰。用 37代毒繁殖生产的三批 CVI988/ Rispens疫苗 ,与国外 CVI988/ Rispens疫苗进行免疫效力比较 ,保护指数为 94 .5 %～ 10 0 %。     

IBV肾型地方分离株弱毒疫苗与同类冻干疫苗的免疫效力试验比较

本试验对人工致弱IBV肾型地方分离株(SF株)与相应进口弱毒疫苗和国产弱毒疫苗进行了免疫效力比较试验。结果表明SF株免疫后产生抗体的时间和抗体效价高低与其他疫苗相当甚至略高,对攻毒鸡的保护率均为100%。剖检实验鸡,采集肾脏样本做肾脏组织切片,观察发现接种致弱的SF株疫苗后,引起的肾脏组织病理变化比选用的2种国产疫苗病理变化轻微,而与进口疫苗引起的病理变化程度相当,属可逆性病变。     

马传染性贫血弱毒疫苗免疫马与自然感染马酶联免疫吸附试验血清抗体耐热性比较研究

本试验对马传贫血清抗体酶联免疫吸附试验(ELISA)的耐热性进行了比较研究.对2匹弱毒疫苗免疫马跟踪检测17个月,在接种3周后ELISA检测均呈阳性,其中一匹马ELISA抗体阳性持续到接种后10个月,另一匹马ELISA抗体阳性可持续到接种后一年.对两匹马的所有不同时期采集的血清样品经75℃加热5 min处理后,用ELISA检测均为阴性.另选6匹马传贫弱毒疫苗免疫马血清且ELISA检测抗体呈阳性的30个样品,经75℃加热5min处理后,均由阳性转变成阴性.4匹经马传贫病毒辽毒株(LN-EIAV)实验感染马且ELISA检测抗体呈阳性的血清样品,经75℃加热5 min处理后,结果仍为阳性.23匹ELISA检测抗体呈阳性的自然感染马血清样品,经加热处理后,19匹马血清样品仍为阳性,4匹马血清样品由阳性转变成阴性.结果表明弱毒疫苗免疫马较自然感染马血清抗体耐热性稍差.     

猪繁殖与呼吸综合征活疫苗(CH-1R株)工厂化生产培养条件的研究

为提高猪繁殖与呼吸综合征病毒(PRRSV)活疫苗CH-1R株的产量和疫苗的质量,本实验探索了PRRSV活疫苗CH-1R株在MARC-145细胞中大规模生产培养的最佳条件。将CH-1R株分别在3L和10L转瓶培养的MARC-145细胞中进行增殖试验,分析CH-1R株在不同接种病毒量和不同接种病毒时间的TCID50变化规律。结果表明,在3L和10L转瓶中,病毒的TCID50最高可达到107.0TCID50/mL以上。CH-1R株通过转瓶在MARC-145细胞中大规模增殖,为PRRSVCH-1R株的工厂化生产奠定了基础。     

高致病性猪繁殖与呼吸综合征活疫苗（JXA1-R株）免疫后在血清中存在时间的报告

本研究应用高致病性猪繁殖与呼吸综合征活疫苗（JXA1－R株）对28～30日龄的仔猪进行免疫,前8周每周采血,8周后每2周采血进行疫苗毒核酸检测,直至免疫后第16周。结果表明,高致病性猪繁殖与呼吸综合征活疫苗（JXA1－R株）在血清中存在的最长时间约为6周,但有的猪呈现间歇性携带疫苗毒。     

Brucella abortus vaccines: comparison of protection provided by immunopotentiated 45/20 bacterins and live strain 19 vaccine in guinea pigs

Guinea pigs were subcutaneously inoculated with 300 microgram of Brucella abortus strain 45/20 killed cells combined in 1% oil emulsion with trehalose dimycolate (TDM), muramyl dipeptide (MDP), or a combination of the 2 immunopotentiators. Protection, as determined by splenic infections in the guinea pigs after challenge exposure, was compared with that induced by strain 19 vaccine. With few exceptions, protection induced by bacterins containing 50 to 1,000 microgram of TDM or TDM-MDP/dose was comparable with that of strain 19 vaccine (P greater than 0.05). Bacterins that contained MDP as an adjuvant were inferior to those with TDM regardless of the excipient or method of preparation. There was no further enhancement of immunogenicity by the addition of MDP to bacterins that already contained TDM. Mineral oil could not be replaced by a metabolizable excipient in bacterins potentiated with TDM.     

免疫不同的IBD中等毒力活疫苗对ND(LaSota)疫苗体液免疫应答影响

维扬麻鸡150只分为5组(即Ⅰ-Ⅴ组).其中I、Ⅱ、Ⅲ组为试验组,对应用市售的A、B、C三种lBD中等毒力活疫苗进行两次免疫接种,Ⅳ组为ND免疫对照组,Ⅴ组为空白对照组,在IBD活疫苗二免后定期监测鸡群免疫ND(LaSota)疫苗后的HI抗体消长情况,以评价这3种IBD活疫苗对NDHI抗体产生的影响程度。结果发现,I组(免疫A疫苗)对NDV的HI抗体水平的影响最大,Ⅱ组(免疫B疫苗)对NDV的HI抗体水平的影响次之,Ⅲ组(免疫C疫苗)对NDV HI抗体水平的影响轻微。