Department of Preventive Medicine, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hnětovínská 3, 775 15, Olomouc, Czech Republic.
Folia Microbiol (Praha). 2012 Sep;57(5):443-6. doi: 10.1007/s12223-012-0149-y. Epub 2012 May 10.
A prerequisite for successful identification of anaerobic pathogenic bacteria from samples of clinical material is the method of cultivation. Currently, several methods of cultivation in anaerobic environment are used: cultivation in anaerobic box, anaerobic jar, and in nonrecurring cultivation system. Here, we determined the suitability of the above methods of cultivation using the estimation of the growth (diameters of colony size) of commonly isolated anaerobic pathogens (Bacteroides fragilis, Clostridium difficile, and Clostridium perfringens). The tested bacterial strains were exposed to atmospheric oxygen for various time periods and then they were cultivated using different anaerobic cultivation systems. Maximum growth differed, depending on the type of cultivation and the strain used. Thus, largest zone diameters, in the majority of measurements, were achieved in the anaerobic box. However, nonrecurring cultivation system seemed better in several cases; this applied to the cultivation of C. perfringens after 15, 30, and 60 min exposure to atmospheric oxygen as well as the cultivation of B. fragilis after 30 and 60 min of oxygen exposure. The cultivation in anaerobic box was the most convenient method for growth of C. difficile. In almost all cases, higher growth was observed in nonrecurring cultivation system than in the system of anaerobic jar. On the other hand, no significant differences were observed among these anaerobic cultivation systems which confirmed their applicability (taking into account some individual features concerning the optimization of cultivations) for identification of pathogenic anaerobes.
成功从临床标本中鉴定厌氧病原菌的前提是培养方法。目前,使用了几种在厌氧环境中的培养方法:厌氧箱、厌氧罐和非循环培养系统。在这里,我们使用通常分离的厌氧病原体(脆弱拟杆菌、艰难梭菌和产气荚膜梭菌)的生长(菌落大小直径)评估来确定上述培养方法的适用性。将测试的细菌菌株暴露于大气氧气中不同的时间段,然后使用不同的厌氧培养系统进行培养。最大生长因培养类型和使用的菌株而异。因此,在大多数测量中,在厌氧箱中获得了最大的区域直径。然而,在某些情况下,非循环培养系统似乎更好;这适用于在大气氧气中暴露 15、30 和 60 分钟后培养产气荚膜梭菌以及在氧气暴露 30 和 60 分钟后培养脆弱拟杆菌的情况。厌氧箱中艰难梭菌的生长最方便。在几乎所有情况下,在非循环培养系统中观察到的生长高于厌氧罐系统。另一方面,在这些厌氧培养系统之间未观察到显著差异,这证实了它们的适用性(考虑到培养优化的一些个别特征)用于鉴定致病性厌氧菌。