Pakpinyo Somsak, Sasipreeyajan Jiroj
Department of Veterinary Medicine, Faculty of Veterinary Science, Chulalongkorn University, Patumwan, Bangkok, Thailand.
Vet Microbiol. 2007 Nov 15;125(1-2):59-65. doi: 10.1016/j.vetmic.2007.05.011. Epub 2007 May 18.
In this study, three consecutive approaches of molecular characterization, determination of minimum inhibitory concentration (MIC) and antimicrobial tested on Mycoplasma gallisepticum (MG) isolated from chicken farms were investigated. These approaches were conducted between 2004 and 2005 to 134 MG samples collected from five different regions of the intensive farming area of Thailand. Twenty MG isolates and four reference strains including S6, F, ts-11, and 6/85 were classified according to Random Amplification of Polymorphic DNA (RAPD) patterns prior to the antimicrobial tests. These isolates exhibited 5 different genotypes (A-E). Consequently, MG isolates representing each genotype were tested on 11 registered antibiotics. The levels of MIC were determined. Three antibiotics, doxycycline (0.20 microg/ml), tiamulin (0.10 microg/ml), and tylosin (0.33 microg/ml), gave the least MICs among all effective drugs. Break point comparisons of each antimicrobial suggested that the MG isolates were most sensitive to lincomycin, oxytetracycline, tiamulin, and tylosin. Some MG isolates had an intermediate effect on josamycin and were resistant to enrofloxacin and erythromycin. Our results also indicated that MG isolated and collected from the region and nearby districts had similar RAPD patterns showing properties of antimicrobial resistance. The RAPD patterns may imply the frequent use of antibiotics and a resistant strain of MG. This is the first report of genetic characterization using RAPD reflected by the levels of MIC against MG. The information is useful to plan for prophylactic and therapeutic impacts on the poultry industry especially in the area of intensive use of antibiotics.
在本研究中,对从养鸡场分离出的鸡毒支原体(MG)进行了分子特征分析、最低抑菌浓度(MIC)测定及抗菌测试这三种连续的方法研究。这些方法于2004年至2005年期间,对从泰国集约化养殖区五个不同地区收集的134份MG样本进行。在抗菌测试之前,根据随机扩增多态性DNA(RAPD)模式对20株MG分离株和包括S6、F、ts - 11和6/85在内的四株参考菌株进行了分类。这些分离株表现出5种不同的基因型(A - E)。因此,对代表每种基因型的MG分离株进行了11种注册抗生素的测试。测定了MIC水平。三种抗生素,强力霉素(0.20微克/毫升)、泰妙菌素(0.10微克/毫升)和泰乐菌素(0.33微克/毫升),在所有有效药物中MIC最低。每种抗菌药物的断点比较表明,MG分离株对林可霉素、土霉素、泰妙菌素和泰乐菌素最敏感。一些MG分离株对交沙霉素有中等作用,对恩诺沙星和红霉素耐药。我们的结果还表明,从该地区及附近地区分离和收集的MG具有相似的RAPD模式,显示出抗菌耐药性特征。RAPD模式可能意味着抗生素的频繁使用以及MG的耐药菌株。这是第一份利用RAPD进行遗传特征分析并通过针对MG的MIC水平反映出来的报告。该信息对于规划对家禽业的预防和治疗影响非常有用,尤其是在抗生素大量使用的地区。