Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
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In Situ Detection and Differential Counts of Bifidobacterium spp. Using Bromocresol Green, a pH-dependent Indicator

  • Kim, Ki-Hwan (Department of Biological Resources and Technology, Yonsei University) ;
  • Shin, Won-Cheol (Department of Bioengineering and Technology, Kangwon National University) ;
  • Park, Young-Seo (Division of Biotechnology, Kyungwon University) ;
  • Yoon, Sung-Sik (Institute of Functional Biomaterials and Biotechnology, Yonsei University)
  • Published : 2007.02.28
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Abstract

The purpose of this study was to develop a simple detection method, possibly at the species-level, that allows for large-scale screening of bifidobacteria. Human fecal samples were plated on MRS-raffinose agar containing cysteine and neomycin sulfate, serving as selective pressure for bifidobacteria, and 0.003%(w/v) bromocresol green. All of the test strains grew well on this medium at $37{\pm}1^{\circ}C$, forming white colonies surrounded by yellow halos, which presented a sharp contrast against the green background. In this disc assay, the required incubation time to develop a yellowish zone varied with the species of Bifidobacterium that was tested, allowing for differential counts and easy identification at the species-level: 10-14 hr for B. bifidum, 20-22 hr for B. catenulatum and B. infantis. and 24-25 hr for B. longum and B. breve. No apparent color was observed for B. angulatum and B. adolescentis 28 hr after inoculation. To evaluate the results of pH indicator-based identification, individual isolates were subjected to a colony-PCR experiment with genus-specific primers. The amplified products from the isolates were in good accordance with those from the reference strains at a level of 95% agreement. These results suggest that the present method could be conveniently applied to cell counts, as well as to the preliminary identification of bifidobacteria from a variety of sample types including human feces, dairy products, and commercial probiotic supplements.

Keywords

References

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