Ethical approval for the feeding study was granted

by Cardiff School of Biosciences, Cardiff University (Approval number 079-1). Cultivation of LAB from faecal samples Fresh faecal samples were weighed, diluted 1:10 MRD diluent (Oxoid, Basingstoke, UK) containing 15% glycerol, and frozen at -80°C; no significant loss of cultivable diversity or viability was observed when freshly resuspended and plated faecal samples were compared to replicate samples that had been stored frozen. buy GW-572016 Serial dilutions were plated in replicate onto MRS and MRS-P agar, incubated at 37°C for 72 hours, and enumerated quantitatively and qualitatively prior to random picking of up to 10 the different colony morphotypes for RAPD fingerprinting. Each serial dilution plate was documented HKI-272 supplier using digital photography; if RAPD detected the presence of either feeding study strain (L. salivarius strain NCIMB 30211 PCI-34051 mw or L. acidophilus strain NCIMB 30156; see Fig. 6), then retrospective counting of all the morphotypes associated with the strains was performed to determine a total count per gram of faeces. Statistical analysis For enumeration of the faecal counts on MRS-P agar, the mean and standard error of the

mean were determined and a 2-sample t-test to compare means (all numerical analysis was performed using MINITAB®

Release 15, Minitab Inc.). The overall results of the Lactobacillus feeding study were analysed non-parametrically using Chi square because of the limited number of subjects and the variables measured. A 2 × 2 data table was constructed for the analysis categorising the data as follows: two columns for the number of volunteers positive and negative for the administered Lactobacillus strains, respectively, and two rows for before and after capsule consumption, respectively (positive cultures Montelukast Sodium for any given volunteer were only counted once). Acknowledgements This work was supported by a grant from the HELP Wales Programme and Cultech Ltd. P.D. acknowledges salary funding from the Wellcome Trust (grant 075586). We thank: Catrin Thomas and Mark Weaver for technical assistance; Martin Day, Peter Vandamme, Julian Marchesi and Nigel Plummer for helpful discussions concerning the manuscript; and Peter Randerson for advice on the statistical analysis. References 1. Reid G: Probiotics and prebiotics – Progress and challenges. International Dairy Journal 2008,18(10–11):969–975.CrossRef 2.