标准化无菌及支原体检验培养基应用试验

经微生物促生长及灵敏度检测合格的无菌及支原体检验培养基各3批,分别在6个兽用生物制品企业进行了应用试验。抽检疫苗52批和半成品抗原5批,3批无菌检验培养基检测结果一致,疫苗2/52批、半成品抗原3/5批污染。禽源支原体检验培养基抽检样品17批、非禽源支原体检验培养基抽检33批及血清支原体检验培养基抽检10批,3批培养基检测结果一致,均无支原体污染。     

国内外兽用生物制品支原体检验方法的比较分析

为探讨优化我国兽用生物制品支原体检验方法的可能性,本文从支原体检验方法、培养基种类及培养条件、培养基质量控制和检验操作方法四个方面,对中国、美国和欧盟兽用生物制品支原体检验方法进行系统比较,分析不同方法之间的差异及优劣。结果发现,与美国《联邦法规》第九卷及《欧洲药典》相比《中国兽药典》支原体检验方法有培养基种类及配方成分全面等方面的优势,但在培养基质量控制方法、检验过程控制、检验标准制定等方面还存在不足。正视这些差距与不足,提出改进和验证的方向,可为我国兽用生物制品支原体检验的优化提供参考。     

支原体检验用培养基的改进

对于支原体检验,最关键的问题是培养基的质量。支原体生长要求的条件非常苛刻,质量好的培养基能使支原体生长滴度达到10-9/ml,反之质量差的只能达到10-1/ml。培养基的质量严重影响着支原体检验的准确性。长期以来许多检验人员对支原体检验培养基的配制方法深感棘手,为此对...     

支原体培养基配制工艺改进试验

根据GMP范要求,对兽医生物制品的疫苗必须进行支原体检测。目前乾元浩南京生物药厂已经可以大量配制用于禽类病毒性活疫苗检验的改良Frey氏液体培养基和固体培养基,以及用于非禽类病毒性活疫苗检验的支原体液体培养基和固体培养基,并对我厂生产的所有兽用疫苗都进行了支原体的检测,结果表明培养基的质量已完全达到了检验的要求;现将其制作工艺与改进方法报道如下。     

猪瘟疫苗质量的替代检验方法研究进展

国标的猪瘟疫苗效力检验、支原体检验、外源病毒检验等质量检验方法均存在试验周期长、操作繁琐、敏感性较低等缺点,随着现代免疫学与分子生物学技术的快速发展,猪瘟疫苗质量检验的新兴替代检验方法日臻发展和完善。论文就近年来针对猪瘟病毒的病毒含量测定、猪瘟疫苗效力检验、支原体检验、外源病毒检验等免疫学与分子生物学新兴检测方法的研究进展做一综述,以期为提高猪瘟疫苗的质量检验水平、保障猪瘟疫苗质量提供参考依据。     

猪支原体肺炎疫苗效力检验常见问题与质量控制

为了更规范、标准地评价猪支原体肺炎疫苗，本研究对近年来猪支原体肺炎疫苗检验进行了系统性分析。结果发现，效力检验作为疫苗质量的核心内容，是检验中最易出现问题参数。对效力检验相关方法、常见问题进行分析，有助于研发过程中规避问题，提高疫苗质量继而保障养殖业发展。     

标准化无菌及支原体检验培养基应用试验

经灵敏度检查合格的标准化无菌及支原体检验培养基已经在4个生物制药公司进行应用实验并且投入使用,分批进行抗原检测,99.5%的培养基经试验检测未被污染,可以投入使用。本文围绕这种无菌及支原体检验培养基进行研究分析,从材料和方法入手,最终进行检测实验以及方法探讨。     

蜂胶佐剂鸡毒支原体灭活疫苗的研制

收获鸡毒支原体培养液,甲醛灭活后加入一定比例蜂胶并进行乳化,制备3批蜂胶佐剂灭活疫苗。3批疫苗的物理性状、无菌检验、甲醛、硫柳汞含量等均符合《中华人民共和国兽用生物制品规程》(2000年版)中同类疫苗的基本要求;安全检验结果表明,受试鸡群无局部和全身反应,体温及采食量正常;效力检验结果表明,在免疫30d后攻毒,3批疫苗的免疫保护率均在60%以上。上述各指标显示,蜂胶佐剂鸡毒支原体灭活疫苗达到国家相关规定,为免疫预防鸡毒支原体病提供了新的方案。     

猪支原体肺炎的诊疗及防治

为了探讨猪支原体肺炎的诊疗和防治，本文从猪支原体肺炎的病原、流行病学、临诊症状、病理变化等方面对该病进行剖析，并阐述诊断该病的检验方法，最后提出该病的防制方法。目的是提高对猪支原体肺炎病的认识，并做好相应的防制措施。     

从进口种猪中检出支原体肺炎病

从进口种猪中检出支原体肺炎病王玉玲朱世强秦贞奎（天津动植物检疫局３００４５７）１９９５年９月，河北某猪场从国外引进约克夏和长白种猪２４０头，在隔离检疫期间，经临床检查、组织病理学检验和血清学检验确诊其中部分猪有支原体肺炎病。现报告如下：１临床症状该批...     

Haemadsorption inhibition test for the identification of Mycoplasma gallisepticum and Mycoplasma synoviae

Summary

Colonies of the avian mycoplasma strains Mycoplasma gallisepticum S6 and Mycoplasma synoviae WVU 1853 and two Mycoplasma synoviae isolates from this laboratory were shown to be haemadsorption positive for chicken erythrocytes. Three Mycoplasma synoviae isolates from this laboratory proved to be haemadsorption and haemagglutination negative. The haemadsorption of the mycoplasma colonies mentioned above was inhibited with specific antisera of either high or low titre. No cross‐inhibition was observed. It is suggested that this test could be used for a quick tentative identification of the two avian mycoplasmas on primary solid‐medium cultures.     

山羊传染性胸膜肺炎支原体C87001株菌种纯化克隆研究

为获得纯净山羊传染性胸膜肺炎支原体生产用菌种,将山羊传染性胸膜肺炎C87001株冻干肺组织（1980年）,通过采用绵羊肺炎支原体代谢抑制试验结合固体培养基挑取单个克隆菌落方法,进行克隆纯化。结果表明,制苗用C87001株冻干肺组织（1980年）纯化后克隆株1～15代菌种均纯净、每代活菌滴度均≥10^9.0CCU／mL,经菌落形态和PCR鉴定,获得克隆纯化的F5代山羊传染性胸膜肺炎支原体生产用菌种。该菌种经本动物回归试验,4／4发病;按规程制备灭活疫苗免疫山羊,攻毒保护为4／4（100％）,对照组3／3发病死亡。     

Ciprofloxacin Treatment Eliminates Mycoplasma in Contaminated Channel Catfish Ovary Cells

Abstract

A temperature-sensitive mycoplasma that was present in the channel catfish ovary (CCO) cell line and in a thymidine kinase-negative mutant of this cell line (CCOBr) was effectively eliminated from the cultures by adding ciprofloxacin, a 4-fluoroquinolone, to the culture medium. Both cell lines have remained mycoplasma free after 3 years of continuous culture. A commercially available genetic probe to mycoplasma ribosomal RNA was used to monitor mycoplasma contamination and evaluate the effectiveness of the treatment.     

Enzootic Pneumonia in Pigs: Propagation of a Causative Mycoplasma in Cell Cultures and in Artificial Medium

Three strains of a new species of mycoplasma were recovered from pneumonic pig lungs, known free of Mycoplasma hyorhinis, by prolonged incubation in pig testicle cell cultures. The three strains produced a characteristic cytopathic effect in the cell cultures. A highly enriched meat-infusion-broth medium was evolved and permitted regular propagation of these organisms. Pneumonia could consistently be produced by intratracheal inoculation of pigs with the mycoplasma propagated in the enriched broth medium or in cell cultures. The mycoplasma were recovered from the lungs of experimentally infected pigs by inoculation into the broth medium. Comparative studies of the pneumonia producing mycoplasma and of M. hyorhinis were carried out in cell cultures, broth media, and in pigs infected experimentally by different routes. The morphological characteristics of the mycoplasma, grown in the different media, are described and illustrated.     

Diagnostic experiences in the routine restrained inspection of turkey stock for mycoplasma infections

A combined culturing medium has been developed and has proved to clearly increase rates of detection of avian mycoplasmas, as compared to mycoplasma culturing media previously used in the GDR. The medium can be used to culture all mycoplasma species relevant to turkey. Experimental studies have shown that pH values should be between 7.0 and 7.2 in liquid culturing media and should not exceed 7.2 in mycoplasma agar, in order to be capable of isolating, in cases of mixed infections, not only Mycoplasma (M.) meleagridis but also M. gallisepticum. Culturing media should be incubated at 38.5 degrees C. The living animal should best be diagnosed by examination of palatine and cloacal swabs, with sperm being additionally checked of insemination cocks. A monitoring programme has been drafted for mycoplasma-free broods of turkey parents.     

绵羊肺炎支原体的培养和PCR鉴定

安徽省3个山羊场有羊只发生肺炎症状,用支原体培养基对病料进行病原分离,经传代和纯化后分离到3株支原体,随后对分离到的支原体进行了形态学观察、PCR鉴定和序列比对,结果显示分离到的支原体均为绵羊肺炎支原体。     

Haemadsorption inhibition test for the identification of Mycoplasma gallisepticum and Mycoplasma synoviae

Summary Colonies of the avian mycoplasma strains Mycoplasma gallisepticum S6 and Mycoplasma synoviae WVU 1853 and two Mycoplasma synoviae isolates from this laboratory were shown to be haemadsorption positive for chicken erythrocytes. Three Mycoplasma synoviae isolates from this laboratory proved to be haemadsorption and haemagglutination negative. The haemadsorption of the mycoplasma colonies mentioned above was inhibited with specific antisera of either high or low titre. No cross-inhibition was observed. It is suggested that this test could be used for a quick tentative identification of the two avian mycoplasmas on primary solid-medium cultures.     

Immunobinding assay for detection of Mycoplasma bovis in milk.

An immunobinding dot-blot assay (IBA) was developed for the detection of mycoplasma in milk. The test was highly species specific when monoclonal antibody preparations were employed in the assay system. Reactions were obtained with all mycoplasma species tested when polyclonal antisera preparations were used. Preincubation for 48-72 hours was necessary with milk samples containing only a few mycoplasma. Time from sample receipt to diagnosis in most positive samples could be reduced from several days by culture to a few hours by the IBA, thus enabling control procedures to be quickly initiated.     

Evaluation of a digitonin disk assay to discriminate between acholeplasma and mycoplasma isolates from bovine milk

A study was undertaken to evaluate effectiveness of a digitonin disk inhibition test to discriminate between Acholeplasma laidlawii and Mycoplasma sp. isolated from bovine milk. The test measured zone diameters of growth inhibition surrounding a digitonin-containing disk on solid medium. Zones of inhibition for 20 isolates of A. laidlawii, ranging from 8-14 mm, did not overlap those of 261 isolates of Mycoplasma sp., ranging from 16 to 38 mm. Examination of variation in zone diameters for M. bovis found that inhibition was not appreciably affected by agar dehydration. Zones of inhibition increased with increasing dilutions of stock culture and decreased with increasing incubation time. Analysis of variance and Fisher's least significant difference test of logn zone diameters revealed that differences in mean logn zone diameters were different at the 0.01 level of significance between some of the six species of mycoplasma examined, indicating that growth among some species of mycoplasma was effected differently by digitonin. The digitonin test was found to clearly discriminate between A. laidlawii and Mycoplasma sp. indicating that the test would be useful as a practical screening test of individual-cow and bulk tank milk for mycoplasmas.