一种用于直接计数以黏土为主的沉积物中细菌的解体方法：溶解硅酸盐及随后对细菌进行荧光染色。

A disintegration method for direct counting of bacteria in clay-dominated sediments: dissolving silicates and subsequent fluorescent staining of bacteria.

作者信息

Boenigk Jens

机构信息

Institute for Limnology, Austrian Academy of Sciences, Mondseestr. 109, A-5310 Mondsee, Austria.

出版信息

J Microbiol Methods. 2004 Feb;56(2):151-9. doi: 10.1016/j.mimet.2003.10.004.

DOI:10.1016/j.mimet.2003.10.004

PMID:14744444

Abstract

The masking of bacteria by abundant microparticles of the clay and silt fraction and cell losses due to sonication hampered direct enumeration of bacteria in sediments dominated by fine sediments. These problems can be circumvented by dissolving silicate fine particles using hydrofluoric acid and subsequent staining of bacteria by DTAF. The developed disintegration method partly replaces mechanical separation of bacteria from sediment particles by chemical disintegration of the silicates. Recovery efficiency ranged from 90% to 111% for different clays and clay-dominated sediments. Especially for the analysis of fine sediments and clays, this method circumvents both strong dilution of the sediment sample and harsh sonication. The method can also therefore be used in sediments where particle abundance is several orders of magnitude higher than bacterial abundance and simple dilution would not suffice in reliably counting bacteria.

摘要

黏土和粉砂级别的大量微粒对细菌的掩盖以及超声处理导致的细胞损失，阻碍了在以细粒沉积物为主的沉积物中对细菌进行直接计数。通过使用氢氟酸溶解硅酸盐细颗粒，随后用二氯三嗪基氨基荧光素（DTAF）对细菌进行染色，可以规避这些问题。所开发的分解方法通过硅酸盐的化学分解部分替代了从沉积物颗粒中机械分离细菌的过程。对于不同的黏土和以黏土为主的沉积物，回收效率在90%至111%之间。特别是对于细粒沉积物和黏土的分析，该方法避免了沉积物样品的强烈稀释和严苛的超声处理。因此，该方法也可用于颗粒丰度比细菌丰度高几个数量级且简单稀释不足以可靠计数细菌的沉积物中。