Department of Animal Health and Antimicrobial Strategies, National Veterinary Institute, Travvägen 20, Uppsala, SE-751 89, Sweden.
Acta Vet Scand. 2012 Sep 21;54(1):54. doi: 10.1186/1751-0147-54-54.
The anaerobic spirochetes Brachyspira hyodysenteriae and Brachyspira pilosicoli cause diarrheal diseases in pigs. Their fastidious nature has hampered standardization of methods for antimicrobial susceptibility testing. For monitoring of antimicrobial susceptibility wild type cutoff values are needed to define where the wild type distribution of MICs ends and no approved cutoffs are available for Brachyspira spp. In this study antimicrobial susceptibility data for both species (in total 906 isolates) were compiled and analyzed and wild type cut off values for B. hyodysenteriae proposed.
The MICs of tiamulin, valnemulin, tylosin, tylvalosin, doxycycline and lincomycin were determined by broth dilution in brain heart infusion broth supplemented with 10% fetal calf serum.
The compiled MICs from the broth dilution tests of the B. hyodysenteriae type strain, B78T (ATCC® 27164T), showed that the method yields reproducible results. In an international perspective the frequencies of isolates with decreased antimicrobial susceptibility were low among both B. hyodysenteriae and B. pilosicoli. However, in B. pilosicoli a constant level of 10-15% isolates with tiamulin MICs >4 μg/ml was detected between 2002 and 2010 and in B. hyodysenteriae a gradual increase in tiamulin MICs was seen between 1990 and 2003 although this increase has ceased during the last years. The wild type cutoff values proposed for B. hyodysenteriae are: tiamulin >0.25 μg/ml, valnemulin >0.125 μg/ml, tylosin >16 μg/ml, tylvalosin >1 μg/ml, lincomycin >1 μg/ml and doxycycline >0.5 μg/ml.
The broth dilution method used in this study has over the years generated tightly grouped MIC populations for the field isolates and reproducible results for the control strain B78T and is therefore a suitable antimicrobial susceptibility test method for monitoring of Brachyspira spp. Here we propose wild type cutoff values for six antimicrobial agents for B. hyodysenteriae tested by broth dilution based on MIC distributions and the current knowledge on mechanisms of resistance in this species. There are few studies on antimicrobial resistance mechanisms and MIC distributions in B. pilosicoli but to some extent the cutoff values proposed for B. hyodysenteriae may be applicable also for monitoring of antimicrobial susceptibility in B. pilosicoli.
厌氧螺旋体巴氏杆菌和猪痢疾密螺旋体引起猪的腹泻病。它们的苛刻性质阻碍了抗菌药物敏感性测试方法的标准化。为了监测抗菌药物敏感性,需要野生型截止值来定义 MIC 野生型分布的终点,并且尚无针对密螺旋体属的批准截止值。在这项研究中,编译并分析了这两个物种(总共 906 株分离株)的抗菌药物敏感性数据,并提出了猪痢疾密螺旋体的野生型截止值。
通过肉汤稀释法在脑心浸液肉汤中测定泰妙菌素、伐奈霉素、泰乐菌素、替米考星、多西环素和林可霉素的 MIC,肉汤中添加 10%胎牛血清。
从猪痢疾密螺旋体 B78T(ATCC®27164T)的肉汤稀释试验中编译的 MIC 表明,该方法可产生可重复的结果。从国际角度来看,两种密螺旋体属(猪痢疾密螺旋体和猪痢疾密螺旋体)的抗菌药物敏感性降低的分离株频率都较低。然而,在猪痢疾密螺旋体中,2002 年至 2010 年间,检测到 10-15%替米考星 MIC 值>4μg/ml 的分离株的恒定水平,而在猪痢疾密螺旋体中,1990 年至 2003 年间,替米考星 MIC 值逐渐增加,尽管近年来这种增加已经停止。建议的猪痢疾密螺旋体野生型截止值为:替米考星>0.25μg/ml,伐奈霉素>0.125μg/ml,泰乐菌素>16μg/ml,替米考星>1μg/ml,林可霉素>1μg/ml,多西环素>0.5μg/ml。
本研究中使用的肉汤稀释法多年来为田间分离株产生了紧密聚集的 MIC 群体,为对照菌株 B78T 产生了可重复的结果,因此是监测密螺旋体属的合适抗菌药物敏感性测试方法。在这里,我们根据 MIC 分布和该物种中耐药机制的当前知识,提出了基于肉汤稀释法检测的 6 种抗菌药物对猪痢疾密螺旋体的野生型截止值。关于猪痢疾密螺旋体的抗菌药物耐药机制和 MIC 分布的研究很少,但对于猪痢疾密螺旋体,提出的截止值在一定程度上也可用于监测抗菌药物敏感性。