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

被引:11
作者
Boenigk, J [1 ]
机构
[1] Austrian Acad Sci, Inst Limnol, A-5310 Mondsee, Austria
基金
奥地利科学基金会;
关键词
disintegration method; sediment; fluorescent staining;
D O I
10.1016/j.mimet.2003.10.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2003 Elsevier B.V. All rights reserved.
引用
收藏
页码:151 / 159
页数:9
相关论文
共 19 条
[1]   FULLY-AUTOMATIC DETERMINATION OF SOIL BACTERIUM NUMBERS, CELL VOLUMES, AND FREQUENCIES OF DIVIDING CELLS BY CONFOCAL LASER-SCANNING MICROSCOPY AND IMAGE-ANALYSIS [J].
BLOEM, J ;
VENINGA, M ;
SHEPHERD, J .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1995, 61 (03) :926-936
[2]   Comparison of detachment procedures for direct counts of bacteria associated with sediment particles, plant litter and epiphytic biofilms [J].
Buesing, N ;
Gessner, MO .
AQUATIC MICROBIAL ECOLOGY, 2002, 27 (01) :29-36
[3]  
Fry J. C., 1988, METHODS AQUATIC BACT, P27
[4]   Optimization of direct cell counting in sediment [J].
Gough, HL ;
Stahl, DA .
JOURNAL OF MICROBIOLOGICAL METHODS, 2003, 52 (01) :39-46
[5]  
Griebler C, 2001, INT REV HYDROBIOL, V86, P453, DOI 10.1002/1522-2632(200107)86:4/5<453::AID-IROH453>3.0.CO
[6]  
2-L
[7]   A TECHNIQUE TO EXTRACT, ENUMERATE AND MEASURE PROTOZOA FROM MINERAL SOILS [J].
GRIFFITHS, BS ;
RITZ, K .
SOIL BIOLOGY & BIOCHEMISTRY, 1988, 20 (02) :163-173
[8]  
Hahn MW, 1998, APPL ENVIRON MICROB, V64, P1910
[9]   Isolation of novel ultramicrobacteria classified as Actinobacteria from five freshwater habitats in Europe and Asia [J].
Hahn, MW ;
Lünsdorf, H ;
Wu, QL ;
Schauer, M ;
Höfle, MG ;
Boenigk, J ;
Stadler, P .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2003, 69 (03) :1442-1451
[10]  
HAHN MW, 1997, TU BRAUNSCHWEIG, V165, P1