A disintegration method for direct counting of bacteria in clay-dominated sediments: dissolving silicates and subsequent fluorescent staining of bacteria.
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.
Boenigk
Jens
J
Institute for Limnology, Austrian Academy of Sciences, Mondseestr. 109, A-5310 Mondsee, Austria. jens.boenigk@oeaw.ac.at
D000538:Aluminum Silicates / Q000737:chemistry*; D001419:Bacteria / Q000302:isolation & purification*; D000077215:Clay; D015169:Colony Count, Microbial / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D019015:Geologic Sediments / Q000382:microbiology*; D006858:Hydrofluoric Acid / Q000737:chemistry; D008856:Microscopy, Fluorescence; D017640:Silicates / Q000737:chemistry*; D012995:Solubility; D013194:Staining and Labeling / Q000379:methods
D000538:Aluminum Silicates; D005456:Fluorescent Dyes; D017640:Silicates; D006858:Hydrofluoric Acid; D000077215:Clay