To assist with selection of the Lactobacillus species in the feeding study, we investigated whether the addition of polymyxin B to MRS medium (MRS-P agar, see Methods) would increase the selectivity of this medium by acting as a counter-selection

against coliforms. Addition of polymyxin B at a concentration of 120 units per ml of agar did not inhibit the viability of any of reference LAB species isolates (Table 2) or the two Lactobacillus strains incorporated into the capsule. However, MRS-P was highly effective at reducing the number of contaminating Gram negative enteric PD-0332991 purchase colonies seen after plating of human faeces. To examine the efficacy of the semi-selective MRS-P developed for enrichment of the LAB species within faeces, 29 Galunisertib solubility dmso of the most dominant cultivable isolates recovered from 10 of the volunteers at days -14, 0 and 28 (before and after Lactobacillus feeding) were randomly selected for molecular identification. Using 16S rRNA gene sequence analysis these dominant isolates were identified as (Table 2; Fig 2): Lactobacillus species (10 isolates), Streptococcus species (7 isolates), Enterococcus species (7 isolates), Weissella species (1 isolate) and Staphylococcus species (4 isolates). The latter Staphylococcus isolates were the only non-LAB species isolated in high numbers on MRS-P agar after faecal plating. These data indicated that

the MRS-P agar was effective for selection of LAB species after faecal culture. Tracking Lactobacillus strains after oral administration RAPD fingerprinting of the major colony morphotypes appearing after cultivation of each faecal sample was used to determine if the Lactobacillus strains had survived gastric and intestinal passage (Fig. 5). The mean faecal LAB count was 8.8 ± 2.7 × 106 cfu per g faeces when all volunteer samples were analysed; consumption

of the lactobacilli did not significantly alter the total faecal LAB counts obtained from any of the volunteers (data not shown). Prior to the start of the study, L. salivarius strain NCIMB 30211, aminophylline was not detected in any of the volunteers, however, strains matching L. acidophilus NCIMB 30156 were cultivated from three of the volunteers at the pre-feeding stage (Table 3). The appearance of this L. acidophilus (RAPD strain type 1; Table 2) at this point in the study was not unreasonable since it appeared to be a strain commonly found in food/probiotic products which may have been consumed by the volunteers (Table 2). Table 3 Detection of Lactobacillus capsule strains and other faecal bacteria during the feeding study Volunteer Detection of strain in faecal samples before and after consumption of the Lactobacillus capsulea Other recurrent strainsb (strains listed in Table 2)   L. salivarius NCIMB 30211 L. acidophilus NCIMB 30156     Before After Before After   Ac – - – + (D7,21,28) 5 strains (L. rhamnosus A+28) Bd – + (D2) – + (D2) 2 strains (S.