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Applied and Environmental Microbiology, November 1999, p. 4949-4956, Vol. 65, No. 11
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Screening of Probiotic Activities of Forty-Seven Strains of Lactobacillus spp. by In Vitro Techniques and Evaluation of the Colonization Ability of Five Selected Strains in Humans

C. N. Jacobsen,1,* V. Rosenfeldt Nielsen,2 A. E. Hayford,1 P. L. Møller,1 K. F. Michaelsen,3 A. Pærregaard,2 B. Sandström,3 M. Tvede,4 and M. Jakobsen1

Department of Dairy and Food Science1 and Research Department of Human Nutrition,3 Royal Veterinary and Agricultural University, Frederiksberg, University Clinic of Pediatrics, H:S Hvidovre Hospital, Hvidovre,2 and Department of Clinical Microbiology, University Hospital of Copenhagen, Copenhagen,4 Denmark

Received 3 June 1999/Accepted 19 August 1999

The probiotic potential of 47 selected strains of Lactobacillus spp. was investigated. The strains were examined for resistance to pH 2.5 and 0.3% oxgall, adhesion to Caco-2 cells, and antimicrobial activities against enteric pathogenic bacteria in model systems. From the results obtained in vitro, five strains, Lactobacillus rhamnosus 19070-2, L. reuteri DSM 12246, L. rhamnosus LGG, L. delbrueckii subsp. lactis CHCC 2329, and L. casei subsp. alactus CHCC 3137, were selected for in vivo studies. The daily consumption by 12 healthy volunteers of two doses of 1010 freeze-dried bacteria of the selected strains for 18 days was followed by a washout period of 17 days. Fecal samples were taken at days 0 and 18 and during the washout period at days 5 and 11. Lactobacillus isolates were initially identified by API 50CHL and internal transcribed spacer PCR, and their identities were confirmed by restriction enzyme analysis in combination with pulsed-field gel electrophoresis. Among the tested strains, L. rhamnosus 19070-2, L. reuteri DSM 12246, and L. rhamnosus LGG were identified most frequently in fecal samples; they were found in 10, 8, and 7 of the 12 samples tested during the intervention period, respectively, whereas reisolations were less frequent in the washout period. The bacteria were reisolated in concentrations from 105 to 108 cells/g of feces. Survival and reisolation of the bacteria in vivo appeared to be linked to pH tolerance, adhesion, and antimicrobial properties in vitro.

* Corresponding author. Mailing address: Royal Veterinary and Agricultural University, Department of Dairy and Food Research, Rolighedsvej 30, 1958 Frederiksberg, Denmark. Phone: 35 28 32 84. Fax: 35 28 32 14. E-mail: Cnj{at}kvl.dk.

Applied and Environmental Microbiology, November 1999, p. 4949-4956, Vol. 65, No. 11
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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