Instituto de Biologia Experimental e Tecnológica (IBET), Av. República, Qta. do Marquês (EAN), 2784-505 Oeiras, Portugal.
Water Res. 2010 Sep;44(17):4850-9. doi: 10.1016/j.watres.2010.07.018. Epub 2010 Jul 24.
A comparison of different isolation techniques and culture media for detection of filamentous fungi and yeasts in the aquatic environment revealed that the use of membrane filtration with the media dichloran rose bengal chloramphenicol (DRBC) optimized fungi detection in terms of abundance and variety in three untreated water sources with very different characteristics (surface water, spring water, and groundwater). The diversity of the fungi population captured by direct DNA extraction of fungi collected by membrane filtration was compared with the isolates obtained after selective growth using different culture media through amplification of the internal transcribed spacer gene and denaturing gradient gel electrophoresis (DGGE). The Czapek-Dox agar, Sabouraud dextrose agar, and DRBC media showed closer similarities to those obtained by the uncultured biomass for the different water sources. Based on these data and the best enumeration results, DRBC is recommended for the assessment of fungi in water sources using culture-based methods. DGGE was also used to monitor temporal variations in the fungal population structure and showed that each water matrix possessed a distinct population profile as well as that changes in the fungal community can be expected in the different matrices throughout the year.
不同的分离技术和培养介质对水生环境中丝状真菌和酵母菌的检测比较表明,在三种未经处理的具有非常不同特性的水源(地表水、泉水和地下水)中,使用膜过滤和含孟加拉玫瑰红氯 霉素(DRBC)的培养基可以优化真菌的检测,提高真菌的丰度和种类。通过对膜过滤收集的真菌进行直接 DNA 提取,对捕获的真菌种群多样性与使用不同培养基进行选择性生长后获得的分离物进行了比较,方法是扩增内部转录间隔区基因并进行变性梯度凝胶电泳(DGGE)。Czapek-Dox 琼脂、沙氏葡萄糖琼脂和 DRBC 培养基与不同水源的未培养生物量获得的结果更为相似。基于这些数据和最佳计数结果,建议在使用基于培养的方法评估水源中的真菌时使用 DRBC。DGGE 还用于监测真菌种群结构的时间变化,结果表明,每种水基质都具有独特的种群特征,并且可以预期真菌群落会在全年不同的基质中发生变化。