番鸭细小病毒和鹅细小病毒二联弱毒细胞苗的研究

应用适应于番鸭胚成纤维细胞(MDEFC)上繁殖,并产生细胞病变的鹅细小病毒(GPV)弱毒疫苗株与番鸭细小病毒(MPV)弱毒疫苗,按适当比例混合,试制了MPV-GPV二联弱毒细胞苗,并测定了各项免疫指标.结果显示试制出的二联苗对1日龄雏番鸭具有良好的安全性和遗传稳定性;免疫后5 d和7 d攻击GPV强毒,保护率分别为75%和100%,同时免疫后7 d血清中MPV-胶乳凝集抑制(LPAI)抗体效价均大于21,21～28 d抗体达高峰,有效免疫期超过60 d.疫苗于-20℃保存期大于12个月.上述结果表明,二联弱毒细胞苗安全有效.     

我国兽用基因工程疫苗商品化进展

对1991-2012年在我国获得注册并取得产品批准文号的兽用基因工程疫进行了分类总结,分析了我国兽用基因工程疫苗商品化现状和存在的主要问题,展望了兽用基因工程疫苗的发展前景.     

H3亚型猪流感病毒分离与鉴定

从东莞和鹤山等地不同猪场采集的40份鼻拭子或病死猪的肺、气管病料中分离到4株有血凝活性的病毒,其中3株病毒与新城疫病毒阳性血清的HI试验为阳性,另外1株病毒与抗猪流感H3亚型猪流感病毒阳性血清的HI试验为阳性;根据猪流感病毒M基因设计引物,扩增出预期的约315 bp片段,表明该病毒为H3亚型猪流感病毒。     

猪繁殖与呼吸综合征病毒双基因多组合DNA疫苗的构建

本实验根据PRRSV结构蛋白的特性和IFNγ及IL-2这两种细胞因子的特性,构建表达PRRSVGP4-GP5、GP5-M、GP5-N、GP5-IFNγ、GP5-IL-2、N-IFNγ、N-IL-2的重组质粒。这些重组质粒在真核细胞中得到了表达。用这些真核表达质粒可以免疫小猪,通过检测其对猪体免疫应答的诱导能力和接种后动物对PRRSV的抵抗力来对基因疫苗的效能进行研究,以期筛选到有效的基因疫苗并为今后进一步的研究提供思路和理论基础。     

Are the Australian poultry industries vulnerable to large outbreaks of highly pathogenic avian influenza?

Objective   To describe the structure of the Australian poultry industry and discuss the potential for highly pathogenic avian influenza (HPAI) to spread between Australian poultry farms.
Procedure   High densities of poultry farms, frequent contacts between farms by service providers, the supply of live poultry markets (LPM) and the presence of free-range duck flocks in affected regions have been identified as risk factors for the spread of HPAI between flocks in outbreaks causing the death or destruction of over 1 million poultry overseas. Data on 1,594 commercial Australian chicken meat, chicken egg, duck and turkey farms were collected by a telephone questionnaire of farm managers to assess the risk of a HPAI outbreak in Australia.
Results and Discussion   Five regions of Australia had farm densities comparable to overseas regions that experienced widespread HPAI. Common service providers routinely contacted different classes and types of farms over wide geographic areas. However, no responding farms supplied LPM and the majority of duck farms did not produce free-range ducks.
Conclusion   Outbreaks of HPAI have the potential to cause serious impacts on the Australian poultry industry. The risk posted by LPM and free-range ducks is limited, but the movement of genetic stock and common service providers could spread infection between companies, industries or geographical regions. Biosecurity measures are therefore considered critical to limit the secondary spread of infection should an outbreak occur.     

Hsp70 enhances presentation of FMDV antigen to bovine CD4+ T cells in vitro

Foot-and-mouth disease virus (FMDV) is the causative agent of a highly contagious acute vesicular disease affecting cloven-hoofed animals, including cattle, sheep and pigs. The current vaccine induces a rapid humoral response, but the duration of the protective antibody response is variable, possibly associated with a variable specific CD4⁺ T cell response. We investigated the use of heat shock protein 70 (Hsp70) as a molecular chaperone to target viral antigen to the Major Histocompatibility Complex (MHC) class II pathway of antigen presenting cells and generate enhanced MHC II-restricted CD4⁺ T cell responses in cattle. Monocytes and CD4⁺ T cells from FMDV vaccinated cattle were stimulated in vitro with complexes of Hsp70 and FMDV peptide, or peptide alone. Hsp70 was found to consistently improve the presentation of a 25-mer FMDV peptide to CD4⁺ T cells, as measured by T cell proliferation. Complex formation was required for the enhanced effects and Hsp70 alone did not stimulate proliferation. This study provides further evidence that Hsp70:peptide complexes can enhance antigen-specific CD4⁺ T cell responses in vitro for an important pathogen of livestock.     

Oral vaccination of brushtail possums (Trichosurus vulpecula) with BCG: immune responses,persistence of BCG in lymphoid organs and excretion in faeces

AIMS: To determine immune responses, and the localisation and persistence of Mycobacterium bovis bacille Calmette-Guérin (BCG) in gut-associated lymphoid tissues (GALT) and other organs in possums vaccinated orally with lipid-formulated BCG vaccine. To determine the duration of excretion and longevity of survival of BCG in the faeces of vaccinated animals.

METHODS: Possums (n=28) were vaccinated with lipid-formulated BCG (1 x 10 ⁸ colony forming units (cfu) of formulated BCG) by the oral route. Control possums (n=17) were fed oral bait pellets containing formulation medium only. Possums were sacrificed at 3 days and at 1, 3, 6 and 8 weeks after vaccination or ingestion of bait. Proliferation responses to bovine purified protein derivative (PPD) were measured in lymphocytes from blood and mesenteric lymph nodes (MLN) and samples of lung, spleen, liver, MLN and Peyer's patches (PP) were cultured for the presence of BCG. The number of BCG organisms excreted in faeces and the duration of excretion were determined in eight vaccinated possums and eight control possums over a 3-week period. In a separate experiment, a further six possums were vaccinated with oral BCG vaccine (5–10 x 10 ⁸ cfu BCG/possum) and their faeces collected over 48–72 h, for culture of BCG. The longevity of survival of BCG in these faeces was determined by storing faecal samples (n=12) under three different conditions: in an incubator (22.5°C), and conditions which simulated the forest floor and open pasture. A proportion (1–2 g) of these faecal samples was collected after storage for 1, 3, 5, 8 or 20 weeks, and cultured for BCG.

RESULTS: Possums vaccinated orally with BCG vaccine showed strong proliferation responses to bovine PPD in peripheral blood lymphocytes at 6–8 weeks post-vaccination (p.v.). Positive lymphocyte proliferation assay (LPA) responses to bovine PPD were first evident in MLN at 3 weeks p.v. BCG was cultured from MLN and PP in a proportion of animals at 3–8 weeks p.v. BCG was not cultured from sections of spleen, lung or liver at any time p.v. BCG was recovered in low to moderate numbers from the faeces of vaccinated possums for up to 7 days, and maximal numbers were cultured in faeces collected 48–72 h p.v. After storage for 1 week, BCG was cultured from all faecal samples placed in the incubator and from a proportion of faeces exposed to conditions similar to those on the forest floor and pasture. With the exception of one faecal sample stored under forest floor conditions which was culture-positive for BCG at 3 and 5 weeks, BCG was not cultured from any other faecal sample stored for more than 1 week.

CONCLUSIONS: Ingestion of oral BCG vaccine by possums was associated with the development of strong cell-mediated immunity in both blood and MLN. Following oral vaccination with BCG, the organisms were localised and persisted in GALT but did not spread to the spleen, liver or lungs. BCG was shed in low to moderate numbers in the faeces for up to 7 days p.v. The viability of BCG excreted in faeces decreased rapidly, particularly when faeces were exposed to an open pasture environment. Oral vaccination of possums with formulated BCG is unlikely to result in undue contamination of the environment with BCG.     

一株鸡大肠杆菌1型及P型菌毛蛋白结构基因的克隆

据国外发表的人病性大肠杆菌１型及Ｐ型菌毛蛋白结构基因（ｐｉｌＡ、ｐａ;ｐＡ）＿序列,设计并合成 于ＰｉｏＡ及ｐａｐＡ保守区的２对引物。经ＰＣＲ从１株带有甘露糖敏感血凝特性（ＭＳＨＡ）及甘露９糖抵抗血凝特性（ＭＲＨＡ）的鸡致病性大肠杆菌（ＨＪ２０ｅ）染色体中扩增到大小分别在６５７ｂｐ、５４ｌｂｐ的２种阳性产物。通过核酸序列测定分别确证所扩增的ＤＮＡ片段分别为ｐｉｌＡ、ｐａｐＡ基因。经酶切、连接、转化     

仔猪水肿病蜂胶佐剂多价灭活苗的研制Ⅰ.疫苗制备工艺

针对仔猪水肿病发病迅速,病死率很高且很难用抗菌药物控制的特点,旨在研制一种预防该病的新型高效疫苗。采用仔猪水肿病多发病型菌株O138,O139,O141以及湖北地方病型菌株作为制苗用菌种,优化其培养条件,采用蜂胶作为制备疫苗的佐剂,对研制的疫苗进行检验。结果表明菌种在优化培养基上生长旺盛,疫苗安检小鼠及仔猪全部健活,效力检验免疫组保护率达到100％。结果表明该疫苗制备工艺切实可行,疫苗安全性好,保护力强,值得进一步研究。