The protoxin Cry1Ac of Bacillus thuringiensis improves the protection conferred by intranasal immunization with Brucella abortus RB51 in a mouse model

Brucellosis is a zoonotic disease affecting many people and animals worldwide. Preventing this infection requires improving vaccination strategies. The protoxin Cry1Ac of Bacillus thuringiensis is an adjuvant that, in addition to increasing the immunogenicity of different antigens, has shown to be protective in different models of parasitic infections. The objective of the present study was to test whether the intranasal co-administration of pCry1Ac with the RB51 vaccine strain of Brucella abortus confers protection against an intranasal challenge with the virulent strain B. abortus 2308 in BALB/c mice. The results showed that co-administration of pCry1Ac and RB51, increased the immunoprotection conferred by the vaccine as evidenced by the following: (1) decrease of the splenic bacterial load when challenged intranasally with the virulent strain; (2) greater in vivo cytotoxic activity in response to the transference of previously infected cells; (3) further proliferation of cytotoxic TCD8+ cells in response to stimulation with heat-inactivated bacteria; (4) increased production of TNF-α and IFN-γ; and (5) significant IgG2a response. These results indicate that the use of the Cry1Ac protein as a mucosal adjuvant via the intranasal route can be a promising alternative for improving current RB51 vaccine against brucellosis.     

布鲁菌S2疫苗株免疫牛与牛种、羊种布鲁菌自然感染牛ELISA鉴别诊断方法的建立

为建立区分猪种布鲁菌S2疫苗株接种奶牛与布鲁菌自然感染奶牛,BLAST比对分析羊种、牛种、猪种、犬种、沙林鼠种和绵羊种6种布鲁菌基因序列,发现repA—related基因是猪种布鲁菌与牛种及羊种布鲁菌的差异基因。设计引物PCR扩增获得repA-related基因片段,克隆并原核表达得到了布鲁菌repA—related融合蛋白,以repArelated蛋白建立间接EI．IsA检测方法。用repA—related蛋白间接ELISA检测猪种s2疫苗株接种动物血清为阳性,检测牛种和羊种布鲁菌自然感染动物血清为阴性。repA—related蛋白间接EusA能从试管凝聚实验（SAT）及常规ELIsA检测阳性的奶牛血清样本中,区分出s2疫苗接种牛与牛种布鲁菌感染牛。     

奶牛四种流产性疾病的流行现状及疫苗研究进展

奶牛布氏杆菌病、传染性鼻气管炎、病毒性腹泻-黏膜病及新孢子虫病是导致我国奶牛流产发病比例升高的主要疾病,有效防控这四种疾病一直是世界关注的重点之一。本文对上述四种疾病的流行现状及疫苗研究进展进行了综述。     

动物布鲁氏菌病疫苗的来历及亚单位疫苗的研究概况

布鲁氏菌病是一种严重危害人和动物健康的人畜共患病,人群感染主要来源于感染的动物及被污染的畜产品,目前疫苗免疫仍是控制动物布鲁氏菌病的主要措施。现有国际参考疫苗主要有牛种疫苗株S19和羊种疫苗株Rev.1,这些疫苗为动物群布鲁氏菌病控制提供重要保障的同时也存在关键的技术瓶颈问题,如安全性差,接种动物出现流产;血清抗体体内持续时间较长,干扰常规诊断等。随着布鲁氏菌基因组测序工作的相继完成及DNA重组技术的发展,新型布鲁氏菌病疫苗的开发成为研究的热点。亚单位疫苗本着其较高的安全性和有效性,在动物群布鲁氏菌病控制过程中具有较大的应用前景。笔者综述了布鲁氏菌病疫苗的发展史,并对布鲁氏菌病亚单位疫苗的研究进展进行概述。     

Tuberculosis and brucellosis prevalence survey on dairy cattle in Mbarara milk basin (Uganda)

We determined the prevalence of tuberculosis and brucellosis reactors in the dairy herds in the Mbarara district of Uganda in 2002. This is one of the most important dairy-production areas of the country and includes both pastoral and agro-pastoral zones. A total of 340 (of 11,995) randomly selected herds were tested for tuberculosis, using the intradermal tuberculosis-skin test and 315 (of 10,562) herds tested for brucellosis using the serum Rose Bengal test.

The herd prevalence for tuberculosis reactors was 74.1% (95% confidence intervals 69, 78), the individual-animal prevalence was of 6.0% (5.6, 6.5) and within-herd range was 1–50% (up to 100% if suspicious reactors were included).

The herd prevalence for brucellosis was 55.6% (50, 61.2) individual-animal prevalence 15.8% (14.8, 16.7) and within-herd range 1–90%.

The reactor prevalence increased with the age of the animals for both tuberculosis and brucellosis.

Tuberculosis reactor prevalences were higher in animals from the agro-pastoral zone. However, the individual-animal and herd prevalences of brucellosis seroprevalences were higher in the pastoral zone.     

正安县山羊主要疫病血清抗体检测报告

为了解正安县山羊主要疫病的发展趋势,通过虎红平板凝集试验、正向间接血凝试验、酶联免疫吸附试验等方法对正安县引进山羊进行布鲁氏杆菌病、O型口蹄疫、AsiaI型口蹄疫、山羊传染性胸膜肺炎、山羊痘等主要疫病血清抗体的检测。结果表明,O型口蹄疫、AsiaI型口蹄疫和山羊痘免疫抗体阳性率分别为79．42％、81．18％和99．58％,未免疫的布鲁氏杆菌病、羊传染性胸膜肺炎的丝状支原体山羊亚种和绵羊肺炎支原体未免疫抗体阳性率分别为0％、0％和3．81％。监测结果为山羊健康的引进、培育和扩繁提供了一些基础数据,对指导养羊生产有一定的意义。     

High protection of mice against Brucella abortus by oral immunization with recombinant probiotic Lactobacillus casei vector vaccine,expressing the outer membrane protein OMP19 of Brucella species

Brucellosis is a zoonotic disease threatening the public health and hindering the trade of animals and their products, which has a negative impact on the economic development of a country. Vaccination is the most effective way to control brucellosis. The recombinant vector vaccines are promising candidates for immunization in humans and animals. In this study, the gene encoding OMP19 antigen was primarily amplified and cloned into an expression vector called pT1NX, and then transformed to L. casei cell via electroporation technique. The expression was confirmed using specific antibody against the recombinant protein via immunological screening tests such as western blot and immunofluorescence assay. Finally, recombinant L. casei was orally fed to mice and the results were further recorded, indicating that the mice group which received OMP19 through L. casei based vaccine represented a very good general and mucosal immune responses protective against challenges with virulent B. abortus 544 strain compared with negative control recipient groups. Therefore, the vaccine produced in this research plan can be a very good candidate for protection against brucellosis.     

Killing of Brucella antigen-sensitized macrophages by T lymphocytes in bovine brucellosis

The present study was an investigation into the role of T lymphocytes in the killing of antigen-sensitized macrophages (MΦ) in bovine brucellosis. Following confirmation of bovine T lymphocyte cell lines derived from Brucella abortus Strain 19 vaccinated steers as antigen-specific in proliferation studies using various antigens, we adapted an apoptosis assay for evaluation of cytotoxicity by these bovine T cells against autologous monocyte-derived macrophages (MDMΦ) as target cells. Various B. abortus antigen preparations were tested including whole γ-irradiated B. abortus bacteria (γBA), a soluble cytosolic protein fraction and a membrane-associated protein fraction. Both polyclonal and cloned T lymphocyte cell lines exhibited cytotoxicity against MDMΦ targets in an antigen-specific fashion. Polyclonal and cloned T lymphocyte cell lines demonstrated cytotoxic responses to varying degrees against B. abortus antigens regardless of whether the antigen used was whole nonviable bacteria, a soluble protein extract or a membrane-associated fraction of extracted bacteria. To further develop correlation of these responses to an in vivo host defense mechanism, cytotoxicity was evaluated using target cells that had been infected with live B. abortus S19 or B. abortus Strain 2308. Cytotoxic responses were also demonstrated consistently against infected targets with either strain of B. abortus although in most cases, cytotoxicity was higher against target cells sensitized with γBA compared to those infected with live bacteria. Cloned T lymphocyte cell lines were all CD4⁺, CD8⁻ cells indicating that the observed cytotoxic responses were most likely due to an inflammatory T_h1 response and may represent an important host defense mechanism induced by vaccination with live attenuated strains of B. abortus in cattle.     

布鲁氏菌病的研究与防控进展

布鲁氏菌病（Brucellosis,简称“布病”）是由布鲁氏菌（Brucella）引起的危害严重的人畜共患病,主要危害人畜生殖系统,人与人之间几乎不传播,患病动物是主要的传染源,近几年疫情急骤回升。作者综述了布病的研究现状及其流行趋势,初步探讨布病综合防控面临的主要困境。     

Seroprevalence of brucellosis in yaks (Poephagus grunniens) in India and evaluation of protective immunity to S19 vaccine

The present study was carried out to explore the seroprevalence of brucellosis in yaks of North-Eastern hilly yak tracts of Arunachal Pradesh, India. Of 374 animals tested, 23.79, 21.11 and 18.98% were found positive for brucellosis using avidin-biotin ELISA (AB-ELISA), Rose-Bengal plate test (RBPT) and standard tube-agglutination test (STAT), respectively. The relative sensitivity and specificity for STAT were 79.77 and 100%, respectively and the same for RBPT were 88.76 and 100%, respectively in comparison to AB-ELISA. The alarming prevalence as recorded was highest among the yak cows (31.42%) followed by heifers (23.85%) and bulls (8.88%). The immune response in yaks following standard dose of calfhood vaccination with Brucella abortus strain 19 vaccine showed that protective antibody level persisted up to 210 days. This is the first report from India on prevalence of brucellosis and immunization with B abortus strain 19 vaccine in yaks. The present investigation would be a valuable guideline for future control measure and eradication programme of brucellosis in yaks.