Gholarezazadeh, Melika

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lA',t'Pi' )V . ,/-t'/t'it'6fsqj rt(tr)uL&(P,4rt,Fi o.r(,i,itr.l,.i,tl ,r*l'.,l.rlS a+l.r $t*.;.r ,:.e i <-t ;LL,,S,},i ,r*, l'.,+. vo;Sr aJ,i,-; -;.r:,llJ:.l:.r:j.ljl ,-y,E3,o3.l.l*.i csl, *ls-r:l ,sss.t o-fi o-lj 3,.8 d.rJ+ .l; -il r#JJis C+sr* +ls-, al::- GJJ ;1 ,ll -il ir*iri J uSiJ*" J l-J 6la ssrc 3l o'ri 151o-11 J1,,,L r" J+sit l+ l$-* rhlR uj ul*,'o.rl_,11;_,;-)li l+L : L. "-fAt*$ t,o;.r--- jS.l :t-iol; .lE -,t\Y11-\Y1V :.J*,--: Jt*,Kerman University of Medical SciencesFaculty of MedicineIn Partial Fulfillment of the Requirements for the Degree Master of Science in:Medical MicrobiologyTitle:Evalution of antimicrobial activitiy of nanosilver particel on pseudomonasaueroginosa isolates recovered from wound and burn infections and analysisof its effect on biofilm formation and expression of rhlR gene during 2015-20t6.By: Melika Gholam rezazadehSupervisor: Dr" Mohammad Reza ShakibaieYear: 20L7ht,ok,,'r)'r U*\' )-,) ,\r."v',f.rbr.,1, ,* * ;i c*9G" ,;l1lil ,[ol: c., .lb,,L* d[6 ,r9re .:l-711 2r ..,,br,:)-l-t o5;9b c*ol oq d+z -ol.rL,*.,) d)d orriu t-i #x,-t*fu;i sF* alx aiy ol1 ;:f hg;g:.; grJ" .l,l jJ.aLl c-l "s .r-,fuL.s c,tJU" 1.: ora; oligtlJ .r-trx "rj ii .?Fl :y l;9419;l u.pUrlrt-, 6F|'r ,-d1)f ,P e;t; ,p> .,: s ./ **Ls ir *l*ix ali- af c,*,! Jis -:j .-(,- 4f rhlR. gi.x tf,.;r; rhlR ".,"i Ok ;lx. x g'i ll .ura; orir;U ,r^.J*i9. s : "o,:.5*" "i; ll ,x "rys d*!,,, gr-! 1r kl .;*.r_fJ-l*( .r; .g;Lls jt",f tl.i .rblL* *3r or;- )\ l;yis;i ,,"Ug):^"* al91.i yA :i.u :b ,-is; : , ya,*y.p19-r g,.,Liy ozii.; ol5pb (MBC) "i.;:, ;l ,rfr:J c&E J,lJ 9 (MIC) fit ) 6#;;:; - - ;*.I"i:" G il s *Lix "j5;:.$,5 e$l oly 6;11,1:rfn Js, + (CLSI) **J!:,ralK,::,"t;i Jj:. --relative quantitative real- .3;(; ily \, rhlR g ih Olx" t!-tej "c.if ehal ;*1"5* ,yc: * c,* : . - -.,o,:l: ,li c')X r)g, o;ii oliltJ 19;> prr 9 )y& r.: time PCR. (qRT-: - -".,,bl"cr9*, 'Jt+.gt" aJgyl t;, )y ,)G 9 c;l.: *.f gfv.l'j il ,;"iU Y. rLl 1.: osii o,:t- --a;l;: ;*.^o ey-b .r,-Li JrS "sl diby" ,-LG ,lq i, 'd,"5 ;li11".* ,ts:>y ,.i:- ;l or^i qg79i eq j .7-_.: -,e rhlR Oj .:'* of JL ,r) "+),F Lr)is^+ ,dq ,rJiX .:""1J c.cL sub-MIC ,:Lls 7.: o2il o1! i- .: .-.c;b r;:11;sl ylX f ,-11,,*" C osil oj:.- .:Lt' u#-k" *^LrX J; 3 G9)S* -r; c_*E .5111,: oy; or:9;[ af .:i.: ;l*:.; dra* O*l e"tI ,-5 y\ 4r! .z;l,c #X ;[*<;; oble; y diX L t :" ur:"lf *J-r .(- Oil- a'rhlR. Oj al11 ,]1.,.r r,r ,gi "Li, .>.3-|,3u ,l c."-l 15^" ",*r .gF-l -r.$J .ita F*St<" Yk>i .c..31.: Ok ",Ll)e! ,xl,x rr Oj y-i e-(;- - .- -.r.i! .r p;Y .gr*.1 c,ult ,ol*iJ*+ J6.$ 1r o;ii o2rgtti il ,t*+K. { Ar"r C, S1X hJ .r:^:L i;- --) -*J ri r#,_t" re.b'"J*,,,or.6.; o;5gL,,ol.'sx J;g4#i.lUrl:rt :rgs.S .t*',r,:#"n: considering the importance of pseudomonas bacteria in the development of Ihospital infections and its increased resistance to multiple drugs (MDRs), the aim of this research Iwas to find an effective way to prevent the formation of biofilms by silver nanoparticles in Ipatients infected wlth Pseudomonas aeruginosa' Although the anti-biofilm effect of silver Inanoparticles has been studied in previous studies, the rerationship between rhlR gene, a control lgene that regulates biofilm levels, has not been reported in the presence ofnano silver' Therefore' ]in addition to the antimicrobial and anti-biofilm effect of silver nanoparticles, its effect on theexpression of rhlRgene expression was also investigated'Methods:23isolatesofpseudomonasaeruginosawereisolatedfromtheburnwardofShafahospital in Kerman. The minimum inhibitory concentration of growth (MC) and minimumbactericidal concentration (MBC) of silver nanoparticles were performed according to the clinicaland laboratory standards institute (cLsD guidelines by broth micro-dilution broth method'Biofilmformationandanti-biofilmeffectofsilvernanoparticleswereperformedbymicrotitermethod.Finally,weexaminedtheexpressionofrhlRgeneexpressionbyquantitativereal-timePCR(qRT-PCR)techniqueinthepresenceandabsenceofsilvernanoparticles.Results: Silver nanoparticles in the 20-nanometer dimension had a strong antimicrobial effectandcouldsignificantlycontrolthegrowthofclinicalisolatesofpseudomonasaeruginosacollected from the healing ward,s burn area. The results also showed that the silver nanoparticlesin the sub-MC concentration reduced the biofilm significantly' while rhlR gene expressionincreased twice in the presence of silver nanoparticles'conclusion:lngeneral,theresultsofthisstudyshowedthatsilvernanoparticlespossessantimicrobial and anti-biofilm properties' Although there was no relationship between theexpressionofrhtRgeneexpressionasabiofilmregulatorandtheabilitytoformbiofilm'itwastwiceaspronouncedforthegene.otherregulatorymechanismsmayalsobeeffectiveinbiofilmformation.Therefore,morestudiesareneededtofindoutthemechanismofsilvernanoparticleeffect in biofilm formation'Keywords:P.aeruginosa,biofilm,nano-Ag,relativequantitativeRT-PCR.

Evalution of antimicrobial activity of nanosilver particel on psudomonas aueroginosa isolates recovered fromwound and burn infections and analysis of its effect on biofilm formation and expression of rhlR gene during ۲۰۱۵-۲۰۱۶

Evalution of antimicrobial activity of nanosilver particel on psudomonas aueroginosa isolates recovered fromwound and burn infections and analysis of its effect on biofilm formation and expression of rhlR gene during ۲۰۱۵-۲۰۱۶

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