The 16MΔvjbR as an ideal live attenuated vaccine candidate for differentiation between Brucella vaccination and infection

Brucellosis brings great economic burdens for developing countries. Live attenuated vaccines are the most efficient means for prevention and control of animal Brucellosis. However, the difficulties of differentiating of infection from vaccine immunization, which is essential for eradication programs, limit their applications. Therefore, the development of a vaccine that could differentiate infection from immunization will overcome the limitations and get extensive application. VjbR is a quorum sensing regulator involving in Brucella's intracellular survival. The vjbR∷Tn5 mutants have been proven effective against wild type strain challenge, implying its possibility of use in vaccine candidate development. To further evaluate this candidate gene, in the present study, the antigenicity of purified recombinant VjbR protein was analyzed. Antibodies to Brucella melitensis VjbR could be detected in sera from patients and animals with brucellosis but not in control ones, implying the potential use of this protein as a diagnostic antigen. Then a vjbR mutant of B. melitensis 16M was constructed by replacing the vjbR with kanamycin gene. The mutant showed reduced survival in macrophage and mice. Vaccination of BALB/c mice with 16MΔvjbR conferred significant protective immunity against B. melitensis strain 16M challenges, being equivalent to which induced by the license vaccine Rev.1. The vjbR deletion mutant elicited an anti-Brucella-specific immunoglobulin G response and induced the secretion of gamma interferon and interleukin-10. The most importance is that, the use of vjbR mutants as vaccines in association with diagnostic tests based on the VjbR antigen would allow the serological differentiation between infected and vaccinated animals. These results suggest that 16MΔvjbR is an ideal live attenuated vaccine candidate against B. melitensis and deserves further evaluation for vaccine development.     

Ghrelin对雌激素诱导雄性小鼠胸腺萎缩的逆转及对相关细胞因子基因表达的影响

SPF级6周龄Balb/c雄性小鼠60只,随机分为3组,每组20只,适应饲养1周后进行动物试验.雌激素+Gh-relin试验组:前2周隔日腹腔注射苯甲酸雌二醇注射液(含苯甲酸雌二醇0.1 mg)0.05 mL/只,后2周按每日腹腔注射Ghrelin(含Ghrelin 60 μg)0.1mL/只;雌激素+生理盐水对照组:前2周隔日腹腔注射苯甲酸雌二醇注射液(含苯甲酸雌二醇0.1mg)0.05mL/只,后2周按每日腹腔注射0.9％生理盐水0.1 mL/只;生理盐水对照组:前2周按0.05 mL/只隔日腹腔注射0.9％生理盐水,后2周按0.1 mL/只每日腹腔注射0.9％生理盐水.动物试验结束后,统计小鼠胸腺指数,应用光镜观察胸腺形态学变化,采用实时荧光定量PCR法检测胸腺中12种细胞因子mRNA表达量的变化.结果显示,注射Ghrelin后,雌二醇诱导的小鼠胸腺萎缩在形态学上基本恢复到正常水平,胸腺中白细胞介素1(IL-1)、IL-2、IL-4、IL-6、IL-12、干扰素(1FN-γ)、白血病抑制因子(LIF)、抑瘤素(OSM)、干细胞因子(SCF)、胸腺体液因子(THF)、肿瘤坏死因子(TNF-α) mRNA含量显著降低(P＜0.05),而IL-7 mRNA含量稍微升高(P＞0.05).结果表明,Ghrelin对雌激素诱导的小鼠胸腺萎缩具有逆转作用,其机制可能是:一方面通过抑制IL一6、OSM、LIF、SCF等细胞因子的表达,从而促进胸腺细胞的增殖;另一方面通过抑制IL-1、IL-2、IL-4、IL-12、THF等细胞因子的表达,从而减少TNF-α和IFN-γ的分泌,进而抑制胸腺细胞的凋亡.     

布鲁氏菌病研究进展

布鲁氏菌病的早期诊断及鉴定是该病防控的重要环节,其诊断技术主要包括细菌学诊断技术、血清学诊断技术以及分子生物学诊断技术。布鲁氏菌病防控应坚持预防为主的策略,其免疫主要以弱毒活疫苗为主,随着分子生物学及重组DNA技术的发展,基因工程疫苗成为近年研究热点,但目前尚未有鉴别疫苗免疫和自然感染的鉴别诊断技术及相应的疫苗制品。本文对布鲁氏菌病的病原学、流行病学、诊断技术和疫苗研究进展等方面进行概述,以期为建立灵敏性高、特异性强、高效便捷且易推广的布鲁氏菌病诊断技术和研发更加安全有效的布鲁氏菌病疫苗提供基础。     

庭院生态养猪新技术

本文介绍了庭院生态养殖的基本内涵，猪舍的构造特点，建筑布局及其应注意的问题，并预测了庭院生态养猪在我国的发展前景。     

诱导猪肺泡巨噬细胞CD163受体表达的细胞因子筛选

猪肺泡巨噬细胞(porcine alveolar macrophages,PAM)中的CD163受体作为明星分子不仅参与正常的生理活动,而且作为重要的病毒受体介导猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)入侵宿主靶细胞,因...     

海南坡鹿迁地保护现状与建议

迁地保护的海南坡鹿种群可以分为野放、半野放和圈养等3种主要类型。野放种群个体数量最多，约占迁地保护种群个体总数的55．3％；其次为半野放种群，其个体数量占35．6％；圈养种群个体数量最少，只占9．1％。半野放是一种较为稳妥的坡鹿种群恢复方式。在海南坡鹿的迁地保护中，应该增加半野放种群个体数量。本文就海南坡鹿的迁地保护提出了7点保护管理及科学研究建议。     

适应无血清培养的HEK293细胞系的培育

为培育适合腺病毒增殖的无血清培养细胞系,本研究采用培养液渐进替换的方法获得了一株适应无血清培养的HEK293细胞(293SF),经检测293SF细胞具有稳定的支持腺病毒增殖与表达外源蛋白的能力;平均病变时间为97 h,比HEK293细胞缩短12.9%;毒价达到107.48TCID50/mL,比HEK293细胞提高43.3%;并且稳定性良好,连续传16代后293SF细胞状态与易感性未发生变化。该细胞系为腺病毒的无血清培养奠定基础。     

Sequence and Phylogenetic Analysis of rDNA ITS of Three Eimeria Species in Rabbit

In order to study whether the internal transcribed spacers (ITS) sequence could be used as a molecular marker for the species identification of rabbit coccidian, the rDNA ITS of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were amplified by polymerase chain reaction (PCR), and were cloned into pGEM-T Easy vector subsequently. The positive recombinant plasmids were identified by PCR and then sequenced. By sequence comparison and comparative analysis with the relative sequences of rabbit Eimeria spp. available in GenBank, the results showed that the lengths of Eimeria intestinalis, Eimeria flavescens and Eimeria magna were 1065, 1009 and 1047 bp, respectively, and the sequence homologies with the same species sequences were 99.2%, 99.0% and 94.5%, respectively, while were 55.3% to 82.1% compared with corresponding sequences of other different species sequences. The phylogenetic analysis using software Mega 5.0 showed that all rabbit coccidia clustered together in a clade, which was divided into two sister lineages, corresponding to the presence or absence of oocyst residuum. The result demonstrated ITS could be used as a molecular marker for the species identification of rabbit coccidia.     

微酸性电解水对受鸡粪液污染鸡蛋表面沙门氏菌的喷雾消毒效果

为验证仅采用微酸性电解水（Slightly Acidic Electrolyzed Water,SAEW）对污染鸡粪液鸡蛋进行喷雾清洗和消毒,能否缓解鸡粪等有机物对消毒效果干扰问题,同时找出最佳有效氯浓度和消毒时间。该研究采用喷雾方式,按先清洗后消毒的流程对比双蒸水（H2O）+H2O组（清洗和消毒都用灭菌过的H2O）、碱性电解水（Electronized Reduced Water,ERW）+ERW组（清洗和消毒都用ERW）、H2O+SAEW组（先用灭菌过的H2O清洗,再用SAEW消毒）、ERW+SAEW组（先用ERW清洗,再用SAEW消毒）、SAEW+SAEW组（清洗和消毒都用SAEW）等方式对污染鸡粪液鸡蛋表面沙门氏菌的杀灭效果,并采用多元非线性回归拟合杀菌模型,评估了有效氯浓度和消毒时间对SAEW+SAEW组和ERW+SAEW组的影响。结果发现,仅采用SAEW对污染鸡粪液的鸡蛋进行喷雾杀菌方式,可有效避免鸡粪液对SAEW杀菌效果干扰,当采用SAEW（ACC (Available Chlorine Concentration)= 25 mg/L）进行喷雾清洗和消毒,清洗时间10 s,消毒时间18 s时,可完全杀灭污染鸡粪液鸡蛋表面沙门氏菌,杀菌值达到6.26 lg cfu/个;SAEW+SAEW组模型决定系数和调整决定系数分别为0.933和0.930,ERW+SAEW组分别为0.926和0.923;验证试验中,SAEW+SAEW组和ERW+SAEW组实际值和预测值的相关系数分别为0.97和0.95;模型成立,该研究可为SAEW在鸡蛋消毒中的应用提供参考。     

赤松纯林胸径结构对枯梢病发生的效应

目的 探究赤松纯林胸径结构对枯梢病发生效应,为合理采取森林经营措施调整林分胸径结构,防控赤松枯梢病的流行提供理论支持和技术支撑。 方法 以昆嵛山赤松纯林为研究对象,设置50块赤松纯林临时样地进行赤松胸径及枯梢病病情调查,应用Weibull分布拟合位置、尺度和形状参数并进行$ {x}^{2} $检验,然后将3参数和赤松平均胸径,感病赤松平均胸径,未感病赤松平均胸径,最大病级赤松平均胸径,最小病级赤松平均胸径表征为赤松胸径特征指标,分别与枯梢病病情指数进行Spearman秩相关性分析。 结果 各赤松纯林临时样地胸径结构均符合Weibull分布,其中胸径分布中心在9.66 ~16.25 cm,90.00%的样地胸径结构表现为正偏山状分布。赤松枯梢病病情指数与未感病赤松平均胸径、最大病级赤松平均胸径和最小病级赤松平均胸径呈负相关,但检验结果为不显著;与尺度参数、赤松平均胸径和感病赤松平均胸径呈显著负相关（p<0.05）,与形状参数呈极显著负相关（p<0.01）;与位置参数呈间接显著负相关。 结论 Weibull分布能够较好地拟合赤松纯林胸径结构。赤松纯林枯梢病病情指数与胸径结构分布中心、胸径结构分布曲线形状、赤松平均胸径和感病赤松平均胸径结构指标存在显著或极显著负相关,与所测赤松最小胸径存在间接显著负相关,可采取抚育间伐等森林经营措施增加林内优势木比重,降低赤松胸径分化程度,减少感病小胸径赤松,达到赤松枯梢病生态调控的目的。