to caspofungin of Candida spp. strains isolated in Ceará, Northeastern Brazil

SHORT

COMMUNICATION/COURTE

COMMUNICATION

Susceptibility

to

caspofungin

of

Candida

spp.

strains

isolated

in

Ceará,

Northeastern

Brazil

Sensibilite

´ a

` la

caspofungine

de

souches

de

Candida

_sp.

_isole

_{´ es}

_en

_Ceara

_´,

Nord-est

Bre

´ silien

C.

Rocha

da

Silva

,

R.

de

Sousa

Campos

,

M.

Adalgiza

dos

Santos

Neta

,

M.

Rozellê

Ferreira

Ângelo

,

H.

Iury

Ferreira

Magalhães

,

B.

Coêlho

Cavalcanti

,

M.

Odorico

de

Moraes

,

D.

Silveira

Macedo

,

H.

Vitoriano

Nobre

Júnior

*

a_{DepartmentofClinicalandToxicologicalAnalysis,SchoolofPharmacy,LaboratoryofBioprospectionand}

ExperimentsinYeast(LABEL),FederalUniversityofCeara´,Fortaleza,CE,Brazil

b_{DepartmentofPathologyandLegalMedicine,SchoolofMedicine,SpecializedMedical,MycologyCenter,}

FederalUniversityofCeara´,Fortaleza,CE,Brazil

c_{CentralPublicHealthLaboratory(LACEN-CE),Fortaleza,CE,Brazil}

d_{DepartmentofPhysiologyandPharmacology,FederalUniversityofCeara´,Fortaleza,CE,Brazil}

Received1August2011;receivedinrevisedform15September2011;accepted23September2011 Availableonline9November2011

KEYWORDS Caspofungin;

Candidaspp.; Microdilution; Susceptibilitytests

Summary Caspofungin isan echinocandinprescribedforthetreatment ofinvasivefungal infectionscausedbyCandidaspp.andAspergillusspp.Theaimofthisstudyistoassessthe degreeofsusceptibilityofCandida spp., isolatedfromblood culturesin thestateofCeará (Brazil)tocaspofunginbythebrothmicrodilutionmethod.Thirty-threestrainsofCandidaspp. wereselectedforthetest(sevenC.albicans,nineC.tropicalisand17C.parapsilosis);these strainsarethemostcommonlyisolatesoffungalinfectionsinCeará.Theresultsofsusceptibility testsbybrothmicrodilutioncanbereadat24or48hoursaftertesting,withoutcompromising testinterpretations.C.parapsilosisexhibitedthehighestMICswhencomparedwiththeMICsof caspofunginagainstC.albicansandC.tropicalis.

#2011ElsevierMassonSAS.Allrightsreserved.

MOTSCLÉS Caspofungine;

Candidasp.;

Résumé Lacaspofungineestuneéchinocandineprescritepourletraitementdesinfections fongiquesinvasivescauséesparCandidasp.etAspergillussp.Lebutdecetteétudeestd’évaluer ledegrédesensibilitéàlacaspofunginedeCandidasp.,isoléàpartird’hémoculturesdansl’état duCeará(Brésil)parlaméthodedemicrodilution.Trente-troissouchesdeCandidasp.ontété

JournaldeMycologieMédicale(2011)21,273—276

* Correspondingauthor.CapitãoFranciscoPedro,1210,CEP-60.420-970RodolfoTeófilo,Brazil. E-mailaddress:helioufc@yahoo.com.br(H.VitorianoNobreJúnior).

Availableonlineat

www.sciencedirect.com

Microdilution; Testsdesensibilité

sélectionnéespourletest(septC.albicans,neufC.tropicaliset17C.parapsilosis);cessouches sontleplussouventisoléesdansdesinfectionsfongiquesenCeará.Lesrésultatsdestestsde sensibilitéparmicrodilutionpeuventêtrelusà24ou48heuresaprèslestests,sans compro-mettrelesinterprétationsdutest.C.parapsilosismontrelesCMIlesplusélevées comparative-mentàcellesdeC.albicansetC.tropicalispourlacaspofungine.

#2011ElsevierMassonSAS.Tousdroitsréservés.

Introduction

Candidaalbicansisthespeciesmostcommonlyisolatedfrom patients with invasive fungal infections (IFIs); however,

C.tropicalisandC.parapsilosisare increasinglyprevalent andaretheprincipalspeciesisolatedinBrazil[8,14,22,16]. Echinocandins are the newest class of antifungal drugs approvedbytheFoodandDrugAdministration(FDA)forthe treatment of systemicfungal infections. Thiswas the first classofantifungalagentstoexhibitactiononthecellwall,by inhibitingtheenzyme(1,3)beta-glucansynthase. Echinocan-dinsexhibitalimitedspectrumofactioncomparedtoazoles. However, they are excellent agents for the treatment of candidiasis.Theprimaryconstituentsofthisgroupare: cas-pofungin,micafunginandanidulafungin[11,3,5,9,18,12].

Thereisnoinvitrosusceptibilitytestingwith echinocan-dins against Candida spp. isolated from blood cultures in Ceará.Theaimofthisstudyistoevaluatethesusceptibility of strains of C. albicans, C.tropicalis and C. parapsilosis

againstcaspofunginbythebrothmicrodilutionmethod,and tocomparetheeffectivenessof caspofunginagainstthese strains.

Materials

and

methods

Isolationandidentificationoffungalstrains

Weused33strainsofCandidaspp.(sevenC.albicans,nine

C.tropicalisand17C.parapsilosis),thestrainswereisolated frombloodcultures,between2009and2010,andarepartof theCollectionofYeastsoftheLaboratoryofBioprospection andExperiments inYeast (LABEL), Departmentof Clinical and Toxicological Analysis, College of Pharmacy, Federal UniversityofCeará.The strainswereinoculatedonpotato agar(HimediaMumbai,India)andincubatedat378C/24h. They were then plated on CHROMagar Candida (Himedia Mumbai,India)toassesspurity.Identificationwasdoneby micromorphologyonriceagarTween80,germtube produc-tion,fermentationandassimilationofcarbohydrates,aswell asmoleculartests[7,2].

Susceptibilitytesting

Caspofungin(Sigma,StLouis,MO)waspreparedinwater,and dilutedinRPMI1640medium(Sigma,St.Louis,MO)buffered to pH 7.0 with 0.165M MOPS (morpholinepropanesulfonic acid)buffer(Sigma).Themicrodilutiontestingwasperformed inaccordancewiththeguidelinesinCLSIdocumentM27-A3 (CLSI2008).MICsweredeterminedvisually,after24hand48h ofincubation,asthelowestconcentrationinmicrogramper millilitreofdrugthatcausedasignificantdiminutionof50%

and90% inhibition of growth belowcontrol levels.Quality controlwas performedbystrainsC. kruseiATCC6258and

C.parapsilosisATCC22019.

Results

and

discussion

Treatmentofsystemicmycosesisoneoftheforemost pro-blemsinthefieldofmedicalmycology;theseinfectionsaffect patientswithpredisposingfactors.Suchpathologiesare diffi-culttodetectandthedelayindiagnosingthemisthereason forhighmortality.Theantifungalagentsavailabletodayoften showsecondaryandsideeffectsbecausetheir therapeutic margin is narrow, which limits usage of such agents[20]. Testing for susceptibility to antifungal agentshas become veryimportantduetotheemergenceoffungiexhibitingsome degreeofresistancetodrugscommonlyusedinclinical med-icine[23,21].InnortheasternBrazil,thereareveryfewstudies thatassesstheincidenceoffungalinfectionsandtheprofileof drugsusceptibility.Whatweobservedinclinicalpractice,ona culturalbasis,wastheprescriptionofantifungaldrugswithout priorsusceptibilitytesting.Thereisanoveralllackoftrained professionalsandreferencelaboratories withqualifiedand knowledgeablepersonnel,and mostimportantly, a lack of integration between laboratories andhospitals. Echinocan-dinsareattractivetherapeuticoptionsforthetreatmentof IFIs. Althoughthese agents have a narrow spectrum, they coverthetwomostcommonIFIs,candidiasisandaspergillosis

[3].Echinocandinsexhibitfewinteractionswithotherdrugs, lowtoxicity,andare broadlyactive againstazole-resistant

Candidaspp.[5].Table1showsthegeometricmean(GM)of MICs50%and90%at24and48hoursofcaspofunginagainst

C.albicans,C.parapsilosisandC.tropicalis.Froma micro-biologicalviewpoint,weseethattheGMofthe50%and90% MICs at 24 and48hours of C. parapsilosisis higher when comparedtotheGMofthe50%and90%MICsat24and48hours ofC.albicansandC.tropicalis;thisdifferencewas statisti-callysignificantwithP<0.05,24and48hoursofcaspofungin vis-a-visC.albicans,C.parapsilosisandC.tropicalis. Epide-miological studiesconducted in different regions of Latin America,includingBrazil,haveindicatedthatC.albicansis themostcommonlyisolatedspecies,followedbyC.tropicalis

andC.parapsilosis[22].C.parapsilosisisoneoftheforemost causesofinvasivecandidiasis[14].Inourstudy,itwasthe principalstrainisolated(17strains)(Table1).Treatmentof IFIsrequiresanunderstandingoftheepidemiologyofspecific infections.Echinocandinsrepresentoneofthemainstaysin thetreatmentagainstcandidiasis,andshouldbemonitoredby identifyingthemostfrequentspecies,aswellasthe suscept-ibilityofsuchspeciestodrugsusedpreviously[5].Inourstudy, theGMoftheMICsofC.parapsilosiscomparedtoC.albicans

and C. tropicalis was higher, showing that C. parapsilosis

exhibitshigherMICsforcaspofungin;similarobservationshave beendescribedintheliterature[9,18,1,4,19].

ThePearsoncoefficient(R)wasusedtoassessthe corre-lationofthelaboratoryresultsat24and48hours.Onecan seeastrongcorrelationbetweentheresultsreadafterthese periodsoftime,suggestingthatthereisnodifferenceinthe release of results at 24 or 48hours aftertesting, thereby speedingupnotonlydiagnosis,butalsoestablishmentofthe appropriate therapy. This coefficient is a measure of the degreeoflinearrelationshipbetweentwoquantitative vari-ables,usedinseveralstudiestocorrelatelaboratory proce-duresandtimeofoperationalization[6,13,17].

Prescribingcaspofunginwithoutpriorsusceptibility test-ingmaycompromisethetreatmentofinfectionscausedby

C.parapsilosis,whichinourstudyandinotherstudieshave been shown to be less susceptible to caspofungin

[9,18,1,4,19]. The union between susceptibility testing andprior identificationof thecausative agentis essential forestablishingthecorrecttreatment.TheGMofthehigh MICsfoundinthestrainsofC.parapsilosiscanbeexplained by the presence of the mutation on thefks genealready reportedinotherstudies.C.parapsilosis,Candida metapsi-losis,and Candida orthopsilosis,which harbor a naturally occurringaminoacidsubstitutionintheequivalentposition ofFks1p.[10,15].Futurestudiesarerequiredtoevaluatethis mutationandtheMICsfound.

Disclosure

of

interest

Theauthorsdeclarethattheyhavenoconflictsofinterest concerningthisarticle.

Acknowledgements

This study was supported byConselho Nacional de Desen-volvimentoCientíficoeTecnológico(CNPq),Coordenaçãode

AperfeiçoamentodePessoaldeNívelSuperior(CAPES)and FundaçãoCearensedeApoioaoDesenvolvimentoCientíficoe Tecnológico(FUNCAP).

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