The MIC was defined as the lowest concentration of antibiotic giving a complete inhibition of visible growth in comparison with inoculated and un-inoculated antibiotic-free wells. Haemolysis test The bacteria were tested for

haemolysis on tryptone soy agar with sheep blood (TSA-SB) (Oxoid Ltd, PB5012A, pH 7.5 ± 0.2, Wesel, Germany) by streaking 24 hr cultures on the blood agar plates followed by incubation at 37°C under anaerobic conditions (Anaerogen, Oxoid) for 24 hrs. The appearance of clear zones around the bacteria colonies indicated the presence of β-haemolysis whereas green zones around the colonies suggested α-haemolysis [42]. Nucleotide accession numbers The nucleotide Anlotinib sequences determined in this study have been assigned GenBank Accession Nos. JQ801703- JQ801728. Results Genotypic characterization The LAB included in the study (Table 1) were isolated from three different African indigenous fermented food products. To confirm their

identities, selected phenotypic tests such as catalase reaction, CO2 production from glucose, colony and cell morphology along with genotypic identification methods were performed. Initially all 33 strains were subjected to rep-PCR (GTG)5 fingerprinting technique for genotypic grouping. Numerical analysis of the (GTG)5-PCR fingerprint band patterns obtained is shown in Figure 1. Figure 1 Dendrogram obtained by A-1210477 cost cluster analysis of rep-PCR (GTG 5 ) fingerprints. The dendrogram is based on Dices’s Coefficient of similarity with the unweighted pair group method with arithmetic averages clustering algorithm (UPGMA). The isolates were identified by 16S rRNA sequencing, IWR-1 in vivo Lb. plantarum group multiplex PCR using recA gene-based primers and W. confusa species-specific PCR method. Sequencing of 16S rRNA gene of all the isolates was performed to further confirm the identities of the strains within each cluster. A BLAST search of the 16S rRNA gene sequences obtained was then performed at NCBI

revealing high similarity values to a number of sequences Protein tyrosine phosphatase in the GenBank database. Strains identified as W. confusa/cibaria showed 99% 16S rRNA sequence homology to both W. confusa and W. cibaria species in the GenBank database. These strains were further subjected to species-specific PCR in order to confirm their true identity. Strains S1 and S2 were previously identified as Lb. paraplantarum based on intergenic transcribed spacers PCR restriction fragment length polymorphism (ITS-PCR/RFLP) grouping, 16S rRNA sequencing and pulsed-field gel electrophoresis (REA-PFGE) [14] and form one cluster group further away from the Lb. plantarum group as shown in the numerical analysis of the (GTG)5-PCR band patterns in Figure 1. However, re-sequencing of the 16S rRNA gene indicated that strains S1 and S2 have high level of sequence homology to both Lb. paraplantarum and Lb. plantarum.