Original paper
New dilution method in microtiter-plates for enumeration and enrichment of copiotrophic and oligotrophic bacteria
Tan, T. L.; Rüger, H.-J.
Archiv für Hydrobiologie Volume 137 Number 4 (1996), p. 511 - 521
33 references
published: Oct 18, 1996
DOI: 10.1127/archiv-hydrobiol/137/1996/511
ArtNo. ESP141013704003, Price: 29.00 €
Abstract
A new microtiter-plate dilution method was applied during the expedition ANTARKTIS-XI/2 with RV "Polarstern" to determine the distribution of copiotrophic and oligotrophic bacteria in the water columns at polar fronts. Twofold serial dilutions were performed with an eight-channel Electrapette in 96-wells plates by mixing 150μl of seawater with 150μl of copiotrophic or oligotrophic Trypticase-Broth, three times per well. After incubation of about 6 months at 2 °C, turbidities were measured with an eight-channel photometer at 405 nm and combinations of positive test results for three consecutive dilutions chosen and compared with a Most Probable Number table, calculated for 8 replicates and twofold serial dilutions. Densities of 12 to 661 cells/ml for copiotrophs, and 1 to 39 cells/ml for oligotrophs were found. Colony Forming Units on copiotrophic Trypticase-Agar were between 6 and 847/ml, which is in the same range as determined with the MPN method.
Keywords
microtiter-plate • copiotrophic • oligotrophic • bacteria