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DIRECT ENUMERATION OF BACTERIA FROM MACROALGAE BY EPIFLUORESCENCE MICROSCOPY AS APPLIED TO THE FLESHY RED ALGAE KAPPAPHYCUS ALVAREZII AND GRACILARIA SPP. (RHODOPHYTA) 1
Author(s) -
Largo Danilo B.,
Fukami Kimio,
Adachi Masao,
Nishijima Toshitaka
Publication year - 1997
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.0022-3646.1997.00554.x
Subject(s) - biology , algae , kappaphycus alvarezii , enumeration , bacteria , botany , dilution , filtration (mathematics) , red algae , serial dilution , staining , seawater , chromatography , ecology , chemistry , medicine , statistics , genetics , mathematics , physics , alternative medicine , pathology , combinatorics , thermodynamics
This report describes a simplified method for direct counting of total bacteria associated with the fleshy red algae Kappaphycus alvarezii (Doty) Doty and Gracilaria spp. A Nuclepore® polycarbonate membrane (0.2–μm pore size) fitted to a vacuum filtration apparatus was used to filter algal tissue homogenate after serial dilution and staining with the fluorochrome 4′,6‐diamidino‐2‐phenylindole. Using epifluorescence microscopy, it is possible to count bacteria without preseparating them from the algae. The technique requires homogenized algal tissue diluted with 0.2‐μm‐filtered, autoclaved seawater to a level appropriate for counting. Dilution reduces the amount of autofluorescent algal debris, which may interfere with Counting. The membrane filtration method yielded a bacterial count two orders of magnitude higher than that of the conventional agarspread plate technique. This method offers a more accurate approach to counting the total number of bacteria on macroalgae.