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Contents
(Abstract) 5
Introduction 6
Materials and methods 8
A. Bacterial strains and culture conditions 8
B. Bacterial biofilm assay to estimate the formation of dental plaque 8
C. Inhibitory effects of water-soluble polymers on the S.mutans biofilm formation 9
D. Thin-layer chromatographic (TLC) analysis of water-soluble polymers 10
E. Hemolysis activity assay on sheep blood agar plates 11
F. Gelatin liquefaction test 11
G. Urease test 11
H. Indole test 11
I. Phenylalanine deaminase test 12
J. Hydrolases assay by TLC 12
K. Enzyme activity associated with production of toxic substances 12
L. Nitroreductase activity 13
M. Antibiotic sensitivity 13
Result 14
A. Inhibitory effects of CMS1 and CMS3 on the formation of S. mutans biofilm 14
B. Analysis of water-soluble polymers produced by the isolates 15
C. Hemolysis activity 15
D. Gelatin liquefaction test 15
E/F. Urease test 16
F/G. Indole test 16
G/H. Phenylalanine deaminase test 16
H/I. Hydrolyses enzyme activity 16
I/J. Enzyme activity test 17
J/K. Nitrate reduction test 17
K/L. Antibiotic sensitivity 17
Discussion 18
Conclusion 22
References 23
(국문초록) 41
Table 1. Effects of W. cibaria and culture supernatant (CS) on the proliferation of S. mutans. 27
Fig. 1. Effects W. cibaria isolates and culture supernatant on the formation of S. mutans biofilm. SM and CS represent S. mutans and culture supernatant, respectively. Error bars represent standard deviations... 28
Fig. 2. Effects of water-soluble polymers of W. cibaria isolates or dextran T-2000 on the formation of S. mutans biofilm. SM represents S. mutans. Error bars represent standard deviations of the mean values of... 29
Fig. 3. Thin-layer chromatogram of products produced by W. cibaria isolates. S, 0.5% sucrose; G, 0.5% glucose; F, 0.5% fructose; 1, the extracts of culture supernatant with 67% ethanol and dried in an 80℃... 30
Fig. 4. TLC pasterns of soluble polymer released from CMU, CMS1, CMS2 and CMS3, those are culture with or without 5% sucrose S; 5% sucrose G; glucose. 31
Fig. 5. Hemolysis activity of W. cibaria isolates. Purified aulture strains (A; CMU B; CMS1 C; CMS2 D; CMS3 E; CCUG) were streaked on 5% blood BAP. After 24hr, the changes of plate color were observed.... 32
Fig. 6. Gelatin liquefaction and degradation by W. cibaria isolates on gelatin agar. W. cibaria (, A; CMU, B; CMS1, C; CMS2, D; CMS3, E;CCUG) (A-E) did not liquity and degrade gelatin agar, whereas P.... 33
Fig. 7. Urea test of W. cibaria isolates on urease agar. W. cibaria (, A; CMU, B; CMS1, C; CMS2, D; CMS3, E; CCUG) (A-E). 34
Fig. 8. Indole test of W. cibaria isolates in trytophan broth. W. cibaria (, A; CMU, B; CMS1, C; CMS2, D; CMS3, E; CCUG) (A-E). 35
Fig. 9. Phenylalanine deaminase that of W. cibaria isolates in phenylalanine slant agar. W. cibaria (, A; CMU, B; CMS1, C; CMS2, D; CMS3, E; CCUG) (A-E). 36
Fig. 10. Hydrolase assay of W. cibaria isolates by thin layer chromatography. W. cibaria CCUG, CMU, CMS1, CMS2, CMS3 (A-E) produced two spot, one spot one is suggested cholic acid (spot 1) and... 37
Fig. 11. Enzyme activity test of W. cibaria isolates by 0.1 M ρ-nitorphenyl-β-D-glucuronide. W. cibaria (, A; CMU, B; CMS1, C; CMS2,D; CMS3, E; CCUG) (A-E). 38
Fig. 12. Detect W. cibaria isolates and E. coli by 4-Nitrobenzoic acid. E.coli showed purple color, however W. cibaria isolates were not changeany color. 39
Fig. 13. Resistance transfer test of vancomycin and chloramphenic from E. faecalis to W. cibaria isolates. W. cibaria CCUG, CMU, CMS1, CMS2, CMS3 (A-E)... 40
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