Matrine Destroy E.Coil Biofilm Formation In Vitro Study

Chapter ?Establish the LB broth-peritoneal dialysis external silicone tube system E.coli biofilm modelObjective:use the LB broth-peritoneal dialysis system external silicone tube to establish E.coli bacterial biofilms(BBF)model in vitro.Method:(1)Congo red agar plates method,preliminary qualitative Clinical strains of whether formation of the BBF(2)using peritoneal dialysis external silicone tube as carrier,and LB broth as a medium build E.coli BBF in vitro models.BBF strains were observed and identified by silver staining and scanning electron microscopes(SEM).Results:(1)standard strain ATCC25922 and the surrounding agar colony Congo red agar plates become black;(2)72 hours later,the dialysis tube visible surface shows a layer of yellow-white mucus membrane-like substance.Carrier of silver staining in ordinary optical microscope can be observed black cotton-like substance;SEM can be observed that E.coli was coral-like aggregates in dialysis tube surface of the bacterial adhesion of how the amount of mucus-like substance a film-like material,forming a three-dimensional structure with a BBF,and the film-like material of uneven thickness.Conclusion:(1)the experimental strain ATCC25922 have the ability to formate BBF;(2)LB broth-dialysis tube system can be establish earlier E.coli biofilm model in vitro in 72 hours.Chapter ?The destructive effect of matrine on E.coli biofilmObjective:To investigate the destructive effects of matrine to the E.coli BBF on the peritoneal dialysis external surface.Methods:using completely randomized controlled design,measure the minimum inhibitory(MIC)of matrine and levofloxacin against E.coli matrine,take one centimeter long each peritoneal dialysis external silicone tube,build E.coliBBF models.The models were randomly divided into the same number of groups,respectively,in three days and seven days is added to the drug intervention.Then randomly divided into four groups within each group,Group A:control group,group B:MIC of levofloxacin(positive control),Group C:MIC matrine,D group:2MIC matrine,E group:4MIC matrine.Each group was divided into 0 hours,4 hours,8 hours,24 hours 4 observation point in time,remove the same number of carriers at each time,rinsed with saline residues of drugs and planktonic bacteria,underwent serial dilution method bacterial colony counts,crystal violet staining semi-quantitative,silver staining and SEM to observe the formation of E.coli BBF.Results:(1)Matrine's MIC is 4.096mg/ml,levofloxacin's MIC is 0.03125ug/ml.(2)Early BBF(3 days):viable count:MIC levofloxacin group,MIC matrine group,2MIC matrine,4MIC matrine are reduced compared with the control colonies,the difference is statistically significant(P<0.05).Crystal violet staining:MIC levofloxacin group,MIC matrine,2MIC matrine,4MIC matrine are reduced compared with the control group,the difference was statistically significant(P<0.05).Silver staining:visible on the ordinary light microscope E.coli BBF control group shows a large dark brown cotton-like growth,with drug intervention of levofloxacin MIC,MIC matrine group,2MIC matrine group,the dark brown cotton-like substance in the 4MIC BBF matrine group is less than the control group.SEM:the control group have a thick accumulation of BBF,and MIC levofloxacin group,MIC matrine group BBF are less than the control group,2MIC matrine,4MIC matrine hardly formate BBF,visible only dispersion of single cells.Mature BBF(7 days)were similar to the earlier BBF.Conclusion:MIC,2MIC,4MIC matrine on early and mature E.coli BBF have destructive effect.