Micin kills the extracellular bacteria although the intracellular bacteria are plated on LB agar plates and cfu enumerated. Reduced invasion (by 40 ) of S. Enteritidis was observed on simultaneous addition with the pathogen along with the probiotic strain within the ratio of 1:1 (Figure 5A). Further, the invasion efficiency of S. Enteritidis was significantly reduced by 80 on addition of KSBT 56 strain 1 h prior to the addition of S. Enteritidis as compared to the control (S. Enteritidis only) (p = 0.0012). Similarly, the invasion of Salmonella was reduced by 23 on co-incubation with CFCS of KSBT 56 strain and by 28 on delayed addition of S.Figure 3 Inhibition of development of S. Enteritidis WT and sodC mutant inside the presence of CFCS (A) of KSBT 56 or reside KSBT 56 (B). A. S. Enteritidis (SEn) WT or even a mutant strain deficient of sodC gene (sodC) were co-incubated with CFCS. B. The above groups were also co-incubated with live KSBT 56 bacterial culture. The cfu was enumerated by plating on LB agar plates supplemented with streptomycin. The presence of CFCS or KSBT 56 is indicated by (+) and also the absence is indicated by (-). The growth of sodC is compared with S. Enteritidis WT strain grown inside the presence of CFCS or reside KSBT 56 strain.Das et al. Gut Pathogens 2013, 5:11 http://gutpathogens/content/5/1/Page five ofFigure 5 Effect of KSBT 56 on invasion of S. Enteritidis (A) and impact of CFCS of KSBT 56 on invasion of S. Enteritidis to HCT-116 cells. A. Gentamicin protection assay was performed to establish the invasion of S. Enteritidis into the HCT-116 cell line in the presence (+) or absence (-) of KSBT 56 strain. The pathogen and also the KSBT 56 strain have been either co-infected together into the cell line (0 h) or the pathogen was added at a time delay of (1 h).1780637-40-2 Order B. The impact of CFCS on invasion of S. Enteritidis was determined by co-incubating S. Enteritidis with all the CFCS of KSBT 56 in 24- properly tissue culture plate seeded with HCT-116 cell line. S. Enteritidis was also cultured with CFCS for 1 h prior to infection of HCT-116 cells. L. plantarum MTCC 1407 was taken as a reference strain. The invasion of S. Enteritidis to HCT-116 cells is taken as manage.Enteritidis after incubation with the pathogen with CFCS for 1 h (Figure 5B). The confocal pictures deliver further conclusive proof from the decreased invasion of S. Enteritidis and adherence on the KSBT 56 strain for the HCT-116 cell line (Figure 6A-D).Adhesion of S. Enteritidis to HCT-116 cell line inside the presence or absence of KSBTProbiotics are known to adhere to intestinal epithelial cells thereby competitively excluding the adhesion of pathogens.1308384-31-7 Formula The adhesion of S.PMID:23075432 Enteritidis to HCT-116 colonDas et al. Gut Pathogens 2013, 5:11 http://gutpathogens/content/5/1/Page six ofFigure six Confocal images of Salmonella invasion (A-D) and Expression of hilA gene by RT-PCR (E). Confocal pictures were taken at 63X magnification working with Leica CLSM. The membrane of HCT-116 cell lines were stained with plasma red dye and S. Enteritidis was tagged with GFP. The KSBT 56 strain was observed in phase contrast. A. S. Enteritidis invasion into HCT-116 inside the absence of KSBT 56 strain. B. S. Enteritidis coinfected with KSBT 56 strain into HCT-116 cell line shows decreased invasion of S. Enteritidis. C. Delayed addition of S. Enteritidis after addition of KSBT 56 strain by 1 h further reduces the invasion of Salmonella into the cell lines. D. Merged image of panel C with phase contrast shows KSBT 56 adhering to HCT-116. E. RT-PCR evaluation of hil.