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1.
The aim of this study was to identify possible effects of different vaccination strategies (concomitantly or not) against brucellosis and clostridia on intake, performance, feeding behavior, blood parameters, and immune responses of dairy heifers calves. Fifty heifers calves were enrolled [38 Gyr (Zebu, Bos taurus indicus) and 12 5/8 Holstein Gyr]. At 120 d of age, animals were randomly distributed among 3 groups: B (n = 18), vaccinated against brucellosis; C (n = 14), vaccinated against clostridia and CB (n = 18), vaccinated concomitantly for both. Rectal and thermographic temperatures were evaluated on days 1, 0, 1, 2, 3, 5, 7,10, 14, and 28 relatives to the vaccination day. Feed and water intake, body weight (BW), and feeding behavior were monitored daily by an electronic feeding system. Blood was sampled on days 0, 3, 7, 14, and 28, relative to the vaccination day for determination of glucose and -hydroxybutyrate (BHBA) concentrations. Blood sampled on day 0 (prevaccination) and on days 28 and 42 were used to evaluate the immune response against Brucella abortus and clostridia. There was an increase in rectal temperature between the first and the third day postvaccination in the 3 groups. The thermography revealed an increase of local temperature for 7 d on groups B and CB. Group C had increased local temperature for a longer period, lasting for up to 14 d. Dry mater intake was reduced for groups B and CB, but no alteration was observed for group C. No alterations regarding initial BW, final BW, average daily weight gain, and feed efficiency were observed. No differences were observed for the 3 vaccination groups for blood parameters throughout the evaluation period. The concomitant vaccination against brucellosis and clostridia led to lower neutralizing antibody titers against epsilon toxin of Clostridium perfringens and botulinum toxin type C of C. botulinum (C > CB > B). When cellular proliferation assay and serological tests to B. abortus were evaluated, no differences were observed between groups B and CB. The present results indicate that the concomitant vaccination against brucellosis and clostridia has no relevant impact on the intake, performance, and feeding behavior of dairy calves. However, the concomitant vaccination of vaccines against these 2 pathogens impacts animal immunity against clostridial infections.  相似文献   
2.
布鲁菌是布鲁菌病的病原体,在世界范围内给养殖业带来巨大损失。自噬是细胞的一种代谢方式,细胞在自噬相关基因的调控下清除细胞内病原微生物和吞噬降解受损、衰老细胞器及大分子物质,以维持机体内环境平衡。布鲁菌入侵细胞后,能够诱发细胞自噬,而这一复杂的过程需要很多细胞因子的参与,探究布鲁菌引发的细胞自噬已成为揭示布鲁菌致病机制的新热点,现就这一新热点的研究进展进行综述。  相似文献   
3.
布鲁氏菌胞内存活机制与巨噬细胞极化关系研究进展   总被引:1,自引:1,他引:0  
布鲁氏菌(Brucella)是一种兼性胞内寄生致病菌,虽无典型的毒力因子却有很强的致病力,且常导致慢性持续感染。布鲁氏菌病被列入世界上严重的人兽共患病之一,直接对畜牧业造成重大经济损失,严重威胁人类健康和公共卫生安全。布鲁氏菌感染的靶细胞主要是巨噬细胞,其发展了更高的策略逃逸宿主免疫细胞的杀伤,甚至在细胞内大量繁殖,削弱巨噬细胞的功能,使巨噬细胞的杀伤作用和抗原递呈功能部分丧失,从而能在宿主细胞内长期持续性感染。文章围绕布鲁氏菌胞内存活机制进行探讨,分析了不同极化类型的巨噬细胞在布鲁氏菌感染过程中的调控作用,以及相关炎症通路对机体炎症发展的作用;揭示了布鲁氏菌胞内生存不仅可适应持续感染期间不同的免疫微环境,也可适应感染期间靶细胞营养物质利用率的差异;证实了在慢性感染的过程中免疫逃避和与宿主细胞代谢的相互作用起关键作用;解释了NF-κB通路是调节M1/M2型巨噬细胞亚型平衡状态的关键因素。布鲁氏菌在宿主细胞中持续感染是国内外学者所面临的巨大难题,其免疫逃逸机制和致病机制仍需进一步研究。  相似文献   
4.
为获得一定纯度及浓度的布鲁氏菌分泌蛋白BspJ并制备多克隆抗体,本研究通过常规PCR方法扩增BspJ基因,连接至pMD19-T克隆载体并筛选阳性克隆;使用限制性内切酶切下目的片段后连接至pET28a(+)载体,双酶切、菌液PCR验证阳性克隆菌并送测序;表达载体pET28a-BspJ转入大肠杆菌DE3后经IPTG诱导表达,收集表达菌进行SDS-PAGE分析;使用His标签蛋白纯化柱纯化目的蛋白rBspJ;将rBspJ蛋白混入弗氏佐剂分4次免疫实验兔,收集兔血清,Western blot鉴定多克隆抗体,饱和硫酸铵法纯化多克隆抗体.结果显示:PCR方法扩增出大小534 bp的目的片段,测序结果正确,表明成功构建出pMD19-T-BspJ克隆载体及pET28a-BspJ表达载体;表达菌表达出大小约24 kDa的rBspJ,纯化后的rBspJ条带明显,基本无杂带,表明成功纯化出rBspJ;真核表达的rBspJ与制备的多克隆抗体反应,表明成功制备多克隆抗体.本研究结果为BspJ蛋白的亚细胞定位分析、功能研究及布鲁氏菌致病机制的研究积累实验数据和奠定基础.  相似文献   
5.
为研究铁调控因子irr和rirA在羊种布鲁氏菌(Brucella melitensis)感染过程中的作用,本研究通过卡那替换的方法构建两个缺失株M5-90Δirr和M5-90ΔrirA,分别将亲本株和缺失株在相同营养条件下培养36 h,观察其振荡培养时的生长变化趋势。分别将1×108 CFU M5-90Δirr、M5-90ΔrirA和M5-90接种到含1.5 mol/L NaCl、pH 2.5、pH 11.5、10 mmol/L H2O2的1 mL布鲁氏菌液体培养基,比较缺失株和亲本株在不同条件下的生长特性;分别以1×106 CFU M5-90Δirr、M5-90ΔrirA和M5-90感染小鼠巨噬细胞RAW264.7检测缺失株的黏附、侵袭和胞内生存能力。结果显示,在相同体外培养条件下缺失株M5-90Δirr和M5-90ΔrirA生长速度明显低于亲本株;M5-90Δirr、M5-90ΔrirA在高盐、强酸和强碱环境下生存率均显著或极显著低于亲本株(P<0.05;P<0.01),在H2O2条件下这两个缺失株的生存率却显著高于亲本株(P<0.05)。与亲本株相比,缺失株在小鼠巨噬细胞RAW264.7内的侵袭和黏附能力均减弱,在感染后45 min缺失株M5-90Δirr和M5-90ΔrirA的黏附和侵袭能力均极显著低于亲本株(P<0.01),但在感染后24 h,这两个缺失株在细胞内繁殖能力与亲本株相比有所增强。本研究报道了irrrirA基因不仅调控了羊种布鲁氏菌的生长,同时对细菌黏附和侵袭能力也产生一定的影响,M5-90Δirr和M5-90ΔrirA是两株具有潜力的布鲁氏菌候选疫苗株。  相似文献   
6.
7.
[目的]探讨采用基因分型方法对非典型布鲁菌鉴定的实用性,为非典型菌株的鉴别提供参考。[方法]采用常规鉴定方法和VITEK 2.0全自动细菌鉴定分析系统对菌株进行初步鉴定,利用AMOS-PCR进行种型鉴定,应用多位点可变数目串联重复序列分析方法(MLVA)确定菌株的基因型。[结果]常规鉴定结果显示试验菌株为疑似布鲁菌;VITEK 2.0全自动细菌鉴定系统显示3株试验菌株均为布鲁菌;AMOS-PCR扩增表明试验菌株均为羊种菌,MLVA聚类分析表明试验菌株与羊种2型布鲁菌紧密地聚为一类,属东地中海基因型,并在Panel 2B中发现了新的基因型(4-4-3-7-5),命名为CN2B-45。[结论]MLVA基因分型方法对非典型布鲁菌具有极高的分辨力,是非典型布鲁菌分型鉴别的最佳策略。  相似文献   
8.
采用热酚水法提取布鲁氏杆菌脂多糖,分离、纯化并SDS-PAGE银染鉴定。纯化的细菌内毒素免疫小鼠,通过斑点ELISA测定血清抗体效价,检测细菌内毒素免疫原性。采用热酚法能获得较高纯度的细菌内毒素,其免疫原性较高,斑点ELISA测血清效价达到1∶1 000 000,与全菌免疫原性接近。本研究为制备布鲁氏杆菌单克隆抗体和建立布鲁氏杆菌病快速诊断方法奠定研究基础。  相似文献   
9.
In this study, we characterized the secreted proteins of Brucella abortus into the enriched media under the bacterial laboratory growth condition and investigated the pathogenic importance of culture supernatant (CS) proteins to B. abortus infection. CS proteins from stationary phase were concentrated and analyzed using 2D electrophoresis. In MALDI TOF/TOF analysis, more than 27 proteins including CuZn SOD, Dps, Tat, OMPs, Adh, LivF, Tuf, SucC, GroEL and DnaK were identified. Cytotoxic effects of CS proteins were found to increase in a dose-dependent manner in RAW 264.7 cells. Upon B. abortus challenge into phagocytes, however, CS proteins pre-treated cells exhibited lower bacterial uptake and intracellular replication compared to untreated cells. Immunization with CS proteins induced a strong humoral and cell mediated immune responses and exhibited significant higher degree of protection against virulence of B. abortus infection compared to mice immunized with Brucella broth protein (BBP). Taken together, these results indicate that B. abortus secreted a number of soluble immunogenic proteins under laboratory culture condition, which can promote antibody production resulted in enhancing host defense against to subsequently bacterial infection. Moreover, further analysis of CS proteins may help to understand the pathogenic mechanism of B. abortus infection and host–pathogen interaction.  相似文献   
10.
A captive adult female bottlenose dolphin presented with stillbirth. The placenta appeared oedematous. No other gross lesions were evident in the placenta or the stillborn calf. Histopathology revealed mild multifocal placentitis and foetal encephalitis. Brucella sp. was isolated from lung, liver, spleen and kidney. Sequence and phylogenetic analysis demonstrated this organism to be most similar to Brucella ceti sequence type (ST) 27. Brucella sp. DNA was detected in formalin-fixed paraffin-embedded placenta and brain by real-time PCR using primers targeting the IS711 gene. Immunohistochemical staining revealed Brucella sp. antigen in placental inflammation. This is the first report of isolation of Brucella sp. from a marine mammal in the Southern Hemisphere and the first report of marine Brucella-associated disease in Australia.  相似文献   
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