Diversity Dynamics of Marine Bacteria Studied by Immunofluorescent Staining on Membrane Filters

Abstract

Of 34 strains of marine bacteria isolated on a general seawater medium, 5 were selected for detailed studies of their population dynamics in the plankton. The isolates were characterized as Aeromonas sp., Chromobacterium cf. lividum, Vibrio sp., and two Pseudomonas spp. Specific antibodies were produced by immunization of rabbits, and bacterial cells were stained on black Uni-Pore membrane filters by an indirect immunofluorescent staining procedure. The method proved to be very specific and practical for use in a large-scale field sampling program. Growth of all five isolates was stimulated by high values for net primary production, chlorophyll a , and dissolved organic carbon. Calculation of a diversity index based on specific and total counts is proposed as a way of characterizing the dynamics of organotrophic bacterial populations in the sea.

Viability of Escherichia coli O157:H7 in natural river water determined by the use of flow cytometry

“…Thus, bacterial cells can be distinguished by 6CFDA-PI double staining and sorted into two populations based on their esterase activity. Sorted bacterial cells can be analysed and identified further at the single cell level by fluorescent antibody staining (Dahle and Laake 1982) or flu-orescent in situ hybridization (FISH) (DeLong et al 1989;Amann et al 1990Amann et al , 1995. Cell growth can indicate active bacterial metabolism as well as respiratory function, enzymatic activity and substrate uptake profile.…”

Utility of Green Fluorescent Nucleic Acid Dyes and Aluminum Oxide Membrane Filters for Rapid Epifluorescence Enumeration of Soil and Sediment Bacteria

“…strain SN45(p111) was enumerated by using an HQ-Cy3 band pass filter set (AHF Analysentechnik, Tübingen, Germany). Immunofluorescence counting was performed as described in the work of Dahle and Laake (2). Total counts were obtained by using SYBR I.…”

Detection, enumeration, and sizing of planktonic bacteria by image-analyzed epifluorescence microscopy

“…epifluorescence microscopy of small, autofluorescent cells contributed to the discovery of the widespread distribution of chroococcoid cyanobacteria in the open ocean (17,35) and is now routinely used for their enumeration (3,20,21). Other recent applications of epifluorescence microscopy include the determination of the frequency of dividing cells as an indicator of instantaneous growth rate (14,22), the use of fluorescent antibodies to detect specific bacterial types (4,7,26,34), and the enumeration and differentiation of chloroplast-containing and apochlorotic nanoplankton populations (5,8,13,27). Biomass estimates of bacterial populations often are made visually by measuring epifluorescent images with an ocular micrometer to determine biovolume and then calculating cell carbon by using appropriate conversion factors (6,9,10,12,22,36,38).…”