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1.
根据培养物的生长速度和含菌量方面筛选适合猪肺炎支原体CJ株生长的猪血清。配制改良Friis培养基时分别添加6种不同生产厂家的猪血清,由6种不同厂家猪血清配制的改良Friis培养基培养猪肺炎支原体CJ株。从生长时间可知,由厂家1的猪血清配制的改良Friis培养基更适宜猪肺炎支原体CJ株的生长。从测定的含菌量测定结果可知,由厂家1的猪血清配制的改良Friis培养基培养猪肺炎支原体CJ株的含菌量较高。结果表明,由厂家1的猪血清配制的改良Friis培养基培养猪肺炎支原体CJ株的生长时间短且含菌量高。  相似文献   

2.
采集疑似猪肺炎支原体病理变化的肺脏,接种到改良牛心培养基、猪胃消化液培养基进行培养,经瑞特染色、镜检,菌落狄氏染色、PCR鉴定及序列比对、生化试验鉴定,抗猪肺炎支原体血清生长抑制试验,分离获得一株猪肺炎支原体云南地方流行菌株,命名为YN200901。试验结果为云南本地株猪肺炎支原体的后续研究提供了物质基础。  相似文献   

3.
从全国部分猪场采集到疑似猪支原体肺炎肺组织病料12份,提取DNA进行猪肺炎支原体PCR和多重PCR检测,将病料研磨后分离猪肺炎支原体,最终分离到1株疑似猪肺炎支原体;通过测序分析、形态观察、生化试验、血清学试验证实其为猪肺炎支原体。该菌株能适应人工培养基的培养,且传代生长良好,液体培养基中培养活菌滴度达109CCU/m L;菌株有一定的致病性,免疫原性好,可作为疫苗备用菌株,该菌株的分离鉴定为研制猪支原体肺炎疫苗奠定了基础。  相似文献   

4.
猪肺炎支原体DJ-166株的分离鉴定   总被引:1,自引:0,他引:1  
从山西某猪场采集疑似猪支原体肺炎肺组织病料,接种CH培养基培养,克隆纯化获得一株菌株。该菌株经PCR、培养特性、生化特性和血清学特性试验鉴定为猪肺炎支原体,命名为DJ-166株。该分离菌株在人工合成培养基上传8代稳定后,活菌计数达到108CCU/m L;菌液培养物气管注射猪后,致病性较弱;免疫原性试验证明该分离菌株具有良好的免疫原性,此结论为猪支原体肺炎疫苗的研究提供了必要条件。  相似文献   

5.
为筛选出山羊支原体山羊肺炎亚种的适宜培养基,将山羊支原体山羊肺炎亚种分离株M1601分别接种Thiaucourt肉汤、MEM-KM2和TSA 3种不同的培养基,测定其在3种不同培养基中生长滴度、生长速度,并测定了M1601在最适培养基中在不同培养阶段的生长滴度。结果表明,添加2g/L丙酮酸钠和150mL/L马血清的Thiaucourt肉汤培养基最适宜山羊支原体山羊肺炎亚种的生长,生长滴度可达109 CCU/mL,在培养6h后开始进入对数生长期,培养60h后进入稳定期,培养72h时后进入衰亡期。此试验结果为山羊支原体山羊肺炎亚种培养特性研究提供了参考数据。  相似文献   

6.
一株猪肺炎支原体HN0613株的分离鉴定   总被引:1,自引:1,他引:0  
采集疑似猪支原体肺炎病理变化的肺脏,经Friis液体、固体培养基培养、纯化,PCR、测序分析、生化鉴定、生长抑制、代谢抑制和致病性试验证实分离获得一株猪肺炎支原体菌株,命名为HN0613.该菌株能适应人工培养基的培养,且传代生长良好,液体培养基中培养达108 CCU/mL;菌株有较强的毒力,可作为疫苗候选株进一步研究.该菌株的分离鉴定为研制猪支原体肺炎灭活疫苗奠定了基础.  相似文献   

7.
绵羊肺炎支原体GH3-3株在改良KM2培养基中的增殖情况   总被引:1,自引:0,他引:1  
绵羊肺炎支原体GH3-3 MoGH3-3株是我国绵羊支原体肺炎灭活疫苗制苗菌株.为研究MoGH3-3株在改良KM2培养基中的生长及繁殖特性,利用活菌计数方法对其在不同培养阶段的生长滴度(颜色变化单位)进行测定,并绘制生长曲线.结果表明,传代稳定的MoGH3-3株在改良KM2培养基中培养12 h开始进入对数生长期,培养7...  相似文献   

8.
为筛选一种适宜绵羊肺炎支原体(Mycoplasma ovipneumoniae,Mo)生长的培养基,利用活菌计数方法对MoGH3-3株在改良KM2培养基、TSB-1培养基、改良Thiaucourt氏培养基和10%马血清改良KM2培养基中不同培养阶段的生长滴度即颜色变化单位(ccu)进行了测定。结果表明,在培养24,48,72和96 h,MoGH3-3株在TSB-1培养基中的平均生长滴度高于其他3种培养基。在培养的72 h和96 h,在TSB-1培养基中平均最大生长滴度达到了109 ccu/mL,在改良KM2培养基和改良Thiaucourt氏培养基中只有108 ccu/mL,而在10%马血清改良KM2培养基中仅为106 ccu/mL。这为绵羊肺炎支原体培养特性研究和疫苗生产工艺研究提供了参考数据。  相似文献   

9.
为了解猪肺炎支原体的培养特性,筛选出肺炎支原体生长的最适培养基,试验采用颜色改变单位(CCU)计数法和PCR方法对猪肺炎支原体在4种常用培养基及改良KM2培养基中生长情况进行了检测与比较。结果显示:改良的KM2培养基为最适培养基,10天时颜色改变单位可达108CCU/mL。  相似文献   

10.
正猪支原体肺炎又称猪气喘病、猪地方性肺炎,是由猪肺炎霉形体(支原体)引起的慢性呼吸道传染病。1流行特点和症状猪支原体是一类无细胞壁的多形态微生物,有球状、环状、点状、杆状和两极状。本菌兼性厌氧,能在无细胞的人工培养基上生长,我国通常采用江苏2号培养基,由于生长条件严格,故一般检验单位不作培养。患肺炎支原体肺炎的患猪表现为呼之困难,肺部前叶有典型性的铅灰色病灶,剧烈的干咳,呼吸粗重,胸部剧烈起  相似文献   

11.
猪支原体肺炎活疫苗(168株)肺内免疫机制研究   总被引:1,自引:0,他引:1  
为研究猪支原体肺炎活疫苗(168株)的免疫机制,通过肺内接种免疫5 ~ 10日龄仔猪,并于免疫后不同时间点检测血清中IgG抗体效价、全血中淋巴细胞转化效率、呼吸道局部的IFN-γ浓度和特异性SIgA滴度,于免疫后28 d剖杀采集呼吸道上皮组织,通过扫描电镜法与原位杂交检测法观察疫苗株在呼吸道的存留以及对纤毛的影响情况.结果发现,免疫后猪血液中淋巴细胞转化增强1.52~2.01倍,支气管表面IFN-γ浓度和特异性SIgA滴度持续增加,但血清抗体一直未检测到.扫描电镜与原位杂交检测结果发现疫苗株能有效地黏附在支气管纤毛上皮细胞上,但对纤毛的影响较小.由此表明,猪支原体肺炎活疫苗(168株)通过肺内免疫可有效激活全身细胞免疫及呼吸道局部的黏膜免疫与细胞免疫反应,而且还可以通过黏附支气管纤毛上皮细胞产生占位效应而对上皮组织不产生损伤.  相似文献   

12.
The present paper compares the fluorescence-serological detection of M. hyopneumoniae and M. hyorhinis in frozen sections with cultural procedures. In 161 lungs of swine the agent of enzootic pneumonia, M. hyopneumoniae, was found almost exclusively in frozen sections (22 times). Only one strain was cultivable. M. hyorhinis was detected 59 times by culture but only 18 times in the sections. For the detection of M. hyopneumoniae therefore the fluorescence-serological investigation of lung sections is the preferable method.  相似文献   

13.
猪胸膜肺炎放线杆菌浊度与细菌计数的关系   总被引:2,自引:0,他引:2  
为了弄清猪胸膜肺炎放线杆菌生长特性、细菌悬液浊度与细菌计数的关系,从而也为探讨细菌致病机理及指导临床上细菌疫苗的生产等提供参考依据,利用地方流行优势菌株APP-7型,将其于TSB液体培养基中培养14 h,按1 h间隔动态取样,测定菌悬液的浊度值(OD630)并用平板法进行活菌计数。结果表明:细菌在接种后2 h进入对数生长期,并持续约3 h,然后进入稳定期。对细菌浊度和细菌计数进行相关性统计分析,发现两变量间存在着极显著相关(P<0.01),以细菌浊度为X,细菌数为Y,建立了曲线回归方程,即:Y=9.817×1014X5.817。  相似文献   

14.
An in vitro culture system for swine tracheal epithelial cells was developed to study the adherence of swine mycoplasmas. Swine tracheal epithelial cells were isolated by enzymatic digestion and cultured on microporous membranes. Growth medium was placed under the membrane support to create air-liquid interface feeding resulting in the cells growing cilia and microvilli on the apical surface. Two strains of Mycoplasma hyopneumoniae (pathogenic strain 91-3 and non-pathogenic type strain J) and two strains of Mycoplasma flocculare (type strain Ms42 and field isolate 7160T) were used in this study. The morphology of the cultured tracheal cells was evaluated by transmission electron microscopy. Adherence of M. hyopneumoniae and M. flocculare and damage to the cilia were demonstrated using scanning electron microscopy. The pathogenic M. hyopneumoniae strain 91-3 adhered to cilia inducing obvious damage. The non-pathogenic M. hyopneumoniae strain J did not adhere to mature cilia. Both M. flocculare strains Ms42 and 7160T adhered to mature and budding cilia. No obvious ciliary damage was observed with strain Ms42. Minimal damage consisting of a slight tangling of the cilia occurred after adherence by strain 7160T. This model will enable us to further study the role of adherence of mycoplasmas on the pathogenesis of swine pneumonia.  相似文献   

15.
The aim of this study was to establish whether enzootic pneumonia could be induced reliably in piglets by administering an aerosolised culture of Mycoplasma hyopneumoniae. Groups of five M hyopneumoniaefree Landrace x Large White piglets weaned at 11 to 14 days of age were exposed to aerosols of in vitro cultures of a virulent strain of M hyopneumoniae. In three separate trials, 14 of 15 pigs exposed to the bacteria developed pneumonia, but pigs exposed to the culture medium alone did not develop the disease. Lung pathology, both gross and histological, indicated acute disease. Ten of the pigs were tested for seroconversion by Western blot and they were all positive. The growth rates of the infected pigs were significantly reduced and the water consumption of the infected groups was also depressed. M hyopneumoniae was recovered from eight of the 15 infected pigs.  相似文献   

16.
几种培养基对A型产气荚膜杆菌产毒效果的比较   总被引:1,自引:0,他引:1  
本文以四种培养基,对同一A型产气荚膜杆菌分离株G3的生长曲线和培养液pH值变化进行了测定,并比较了四种培养基的产毒效果。结果表明:A型产气荚膜杆菌在四种培养基上的生长内线基本一致,但pH值变化有差异,其中以疱肉培养液的pH值变化幅度最小,且其毒素产量最多,活性最高。由此可见,疱肉培养基可作为A型产气荚膜杆菌的产毒培养基。  相似文献   

17.
肺炎支原体的培养和保存条件非常苛刻,是疫苗规模化生产的工艺难题。本文针对猪支原体肺炎活疫苗(168株)生产工艺关键技术进行研究。通过培养试验筛选四种培养基配方表明该疫苗株在低血清改良培养基中生长良好;优化发酵培养工艺,使其在发酵罐培养60~70 h的峰值可达到1010CCU/mL;设计筛选该疫苗耐热保护剂和冻干工艺,37℃下保存10 d的耐老化试验结果显示,下降滴度小于100.5CCU/mL。本研究为提供高效、安全和稳定的猪肺炎支原体疫苗产品奠定基础。  相似文献   

18.
The protective activity of Mycoplasma hyopneumoniae inactivated vaccine prepared from sedimented whole cells and cell-free culture supernates was evaluated experimentally using hysterectomy-produced, colostrum-deprived pigs in which mycoplasmal pneumonia had been induced. The culture supernate vaccine containing less than 10(1) colour-changing units (CCU)/0.2 ml of M. hyopneumoniae significantly (P < 0.05) reduced the percentage of lung lesions compared to controls (3.2 +/- 3.9 vs. 12.2 +/- 2.2%), whereas the sedimented whole cells vaccine containing 10(10) CCU/0.2 ml of organisms provided variable protection (18.7 +/- 16.5 vs. 12.2 +/- 2.2%). Serum from the pigs vaccinated with culture supernate reacted with six protein bands of 97, 89, 65, 46, 42 and 41 kDa by immunoblot analysis. From these results, we conclude that vaccination with culture supernate of M. hyopneumoniae can provide protection against M. hyopneumoniae infection and that these antigens in the culture supernate may be closely related to the reduction of lung lesions.  相似文献   

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