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Applied and Environmental Microbiology, September 2003, p. 5731-5735, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5731-5735.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Selective Plating Underestimates Abundance and Shows Differential Recovery of Bifidobacterial Species from Human Feces

Juha H. A. Apajalahti,1* Anu Kettunen,1 Päivi H. Nurminen,1 Hanna Jatila,1,{dagger} and William E. Holben2

Enteromix Research, Danisco Innovation, FIN-02460 Kantvik, Finland,1 Microbial Ecology Program, Division of Biological Sciences, University of Montana, Missoula, Montana 598122

Received 21 February 2003/ Accepted 19 June 2003

The aim of the present work was to compare the efficacies and levels of selectivity of different culture-dependent and -independent methods for analyzing bifidobacteria in human stool samples. The three different culture media used here significantly differed from each other, particularly with regard to the recovery of Bifidobacterium adolescentis. Bifidobacterium medium failed to recover B. adolescentis; Beerens medium recovered some B. adolescentis organisms (17% of total bifidobacteria), whereas tomato-Eugon medium recovered mainly B. adolescentis organisms (58% of total bifidobacteria). A culture-independent method that combines GC fractionation of bacterial community DNA and 16S rRNA sequencing indicated that B. adolescentis organisms accounted for 85% of all bifidobacteria. Methodological biases, such as those described in this paper, should be taken into account in interpreting earlier studies and designing future experiments.

* Corresponding author. Mailing address: Danisco Innovation, Sokeritehtaantie 20, FIN-02460 Kantvik, Finland. Phone: 358-400-307257. Fax: 358-9-2982203. E-mail: juha.apajalahti{at}danisco.com.

{dagger} Present address: Valio Ltd./R&D, FIN-00039 Helsinki, Finland.

Applied and Environmental Microbiology, September 2003, p. 5731-5735, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5731-5735.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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