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Applied and Environmental Microbiology, September 1998, p. 3246-3255, Vol. 64, No. 9
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Rapid Determination of Bacterial Abundance, Biovolume, Morphology, and Growth by Neural Network-Based Image Analysis

Nicholas Blackburn,1,* Åke Hagström,2,dagger Johan Wikner,3 Rocio Cuadros-Hansson,3 and Peter Koefoed Bjørnsen2

Marine Biological Laboratory, DK-3000 Helsingør,1 and National Environmental Research Institute, DK-4000 Roskilde,2 Denmark, and Umeå Marina Forskningscentrum, Norrbyn, S-910 20 Hörnefors, Sweden3

Received 20 November 1997/Accepted 27 May 1998

Annual bacterial plankton dynamics at several depths and locations in the Baltic Sea were studied by image analysis. Individual bacteria were classified by using an artificial neural network which also effectively identified nonbacterial objects. Cell counts and frequencies of dividing cells were determined, and the data obtained agreed well with visual observations and previously published values. Cell volumes were measured accurately by comparison with bead standards. The survey included 690 images from a total of 138 samples. Each image contained approximately 200 bacteria. The images were analyzed automatically at a rate of 100 images per h. Bacterial abundance exhibited coherent patterns with time and depth, and there were distinct subsurface peaks in the summer months. Four distinct morphological classes were resolved by the image analyzer, and the dynamics of each could be visualized. The bacterial growth rates estimated from frequencies of dividing cells were different from the bacterial growth rates estimated by the thymidine incorporation method. With minor modifications, the image analysis technique described here can be used to analyze other planktonic classes.

* Corresponding author. Mailing address: Marine Biological Laboratory, Strandpromenaden 5, DK-3000 Helsingør, Denmark. Phone: 45 49211633, ext. 326. Fax: 45 49261165. E-mail: mblnb{at}mail.centrum.dk.

dagger Present address: Kalmar University, S-39129 Kalmar, Sweden.

Applied and Environmental Microbiology, September 1998, p. 3246-3255, Vol. 64, No. 9
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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