Study On The Inhibitory Effect Of Anti-streptococcus Mutans-IgY On Streptococcus Mutans

Objective Streptococcus mutans is one of the main pathogens of rickets.The yolk antibody is an antibody produced by avian specific antigen,which is produced by B lymphocytes and transferred to the yolk.The anti-Streptococcus mutans antibody is prepared and purified;Determination of OD value,extracellular polysaccharide and glucosyltransferase activity of S.mutans-IgY against S.mutans,biofilm formation and activity effects.Studying the antibacterial mechanism against S.mutans-IgY provides a more favorable theoretical basis for the anti-caries properties of yolk antibodies.Anti-S.mutans-IgY is a new type of drug with stable,safe and non-resistance characteristics,which provides a new idea for clinical prevention and treatment of dental caries.Methods The formaldehyde-inactivated S.mutans was immunized to the 150-day-old white-air hens 4 times at intervals of 10 days.Eggs were extracted by two-step ammonium sulfate precipitation method,and the titer of the immunization was detected by ELISA.Determination of anti-S.mutans-IgY concentration after purification by BCA method.S.mutans were cultured in different concentrations of egg yolk antibody,and cultured at 0 h,3 h,6 h,9 h,12 h,15 h,18 h,21 h,24 h,and 27 h,respectively,using an ultraviolet spectrophotometer,selecting a wavelength of 630 nm and measuring each The bacterial OD value of the time period and plot the growth inhibition curve.The indolone method was used to determine the water-insoluble and water-soluble extracellular polysaccharides,the Neson-Somogyi method was used to determine the activity of GTF enzyme,Scanning electron microscopy for plaque biofilm morphology.The plaque biofilm after 24 hours was observed by scanning electron microscopy,andthe cross-linking between bacteria was tight and densely cross-linked into a network structure.The XTT reduction colorimetric method was used to study the effect of IgY on the total amount of biofilm.The plaque biofilm was observed by laser confocal microscopy.The composition of bacteria and live bacteria..All experimental data were analyzed using SPSS 16.0 statistical software.Results The highest titer of specific IgY after ELISA was 1:256000 and maintained for 40 days.After purification by BCA method,IgY is converted into 1g egg yolk by two steps of ammonium sulfate precipitation to obtain total IgY about 13 mg.With the increase of IgY concentration,the OD value of bacteria decreased at 24 h,and the growth curve of 4 mg/L and 8 mg/LIgY group was gentle and low.In the inhibition of acid production,1g/L,2mg/L,4mg/L and 8mg/L four groups of IgY could inhibit the acid production of Streptococcus mutans,and the difference was statistically significant.The ability of IgY to synthesize water-insoluble extracellular polysaccharides by S.mutans was significantly inhibited compared with the control group;the reducing sugar and enzyme activity of S.mutans decreased after drug treatment;the XTT method showed that the inhibition rate of IgY to S.mutans biofilm was 17.02%.~74.13%;CLSM showed that with the increase of the concentration of the drug solution,the green viable bacteria and agglomerate structure decreased,and the biofilm thickness was significantly reduced.Conclusions Anti-S.mutans-IgY can effectively inhibit the growth of Streptococcus mutans,acid production,scorpion virulence factors and plaque biofilm activity.A certain concentration of IgY can inhibit the proliferation of S.mutans and has anti-caries effect.