bartonella quintana detection in demodex from erythematotelangiectatic rosacea patients

Short CommunicationNathalia Murilloa, Oleg Mediannikova, Je´rome Aubertb, Didier Raoulta,* a Unite´ des Rickettsies CNRS UMR 6020, IFR 48, Faculte´ de Me´decine de Marseille, Universite´

Short Communication

Nathalia Murilloa, Oleg Mediannikova, Je´rome Aubertb, Didier Raoulta,*

a Unite´ des Rickettsies CNRS UMR 6020, IFR 48, Faculte´ de Me´decine de Marseille, Universite´ de la Me´diterrane´e, 27 Bd Jean Moulin,

13385 Marseille cedex 5, France

b

Galderma R&D, Sophia Antipolis, SNC Les Templiers, Biot, France

1 Introduction

Bartonellaquintanaisknownasthecauseoftrenchfever,butit

may also cause chronic bacteraemia and/or endocarditis and

bacillaryangiomatosis.Theinfectionisusuallytransmittedbylice

in homeless people.1 Bartonella species have been reported in

many other arthropods, including mites2,3 and soft and hard

ticks.4,5Interestingly,thetransmissionofB.quintanaby

Derma-nyssus sp in family members was recently reported.2 Here we

reporttheserendipitousdetectionofB.quintanainDemodexmites

fromapatientwitherythematotelangiectaticrosacea

2 Methods

Forty-fivesubjectsfromthegeneralpopulationofSchenefeld/

Hamburgandneighbouringcommunitieswererecruitedintothis

study,withtheallocationof15subjectsperstudygroupasfollows:

subjectswitherythematotelangiectaticrosacea,withflushingand

persistent central facial erythema and optional telangiectasia

(meanage54.8910.49years;n=15)andsubjectswith

papulo-pustular rosacea defined as moderate to severe with persistent

centralfacialerythemaandtransientpapulesand/orpustules(mean

age50.8611.2years;n=15),fromtheGaldermarosaceadatabase; and 15 healthy subjects with no active skin disease (mean age 52.3813.68years).Subjectswithrosaceareceivednotreatment andwereaskedtostopanytreatmentwithbenzoylbenzoate,lindane, pyrethrin, malathion, allethrin, crotamiton, and metronidazole at least7daysbeforesampling

Demodexweresampledbystandardized skinsurfacebiopsy from the alar crease.6 A total of 356 Demodex mites from

28patientswereselectedundertheloop(115magnification)and werewashedin70%ethanol.Inordertoidentifythebacterialflora

of the mites, DNA was extracted from individual mites by a modifiedprotocolwiththeQIAmpDNAMiniKit(Qiagen).Finally, thebacterialcollectionof72Demodexmitesfrom13patientswas assessedby16SrDNAclonelibraries

3 Results

Weweresurprisedtoidentifythe16SrDNAofB.quintanain twoDemodexmitesfromthesamepatient.Amplificationofspacer

8947matchedwithB.quintanaToulousestrain(coverage100%and identity100%,GenBankaccessionnumberAY660712)

We developed a specific qPCR targeting the vompD gene sequence (AY618465.1) with forward 50 -GGGTGAGGTTAG-CATCGGTA-30,reverse50-TTGACTGCATCGGTGTCTTC-30,andprobe FAM-AGCCTGCTGCCACTCCTGCT We tested all collected mites

A R T I C L E I N F O

Article history:

Received 2 June 2014

Received in revised form 25 July 2014

Accepted 28 July 2014

Corresponding Editor: Eskild Petersen,

Aarhus, Denmark

Keywords:

Bartonella quintana

Rosacea

Demodex

S U M M A R Y

http://creativecommons.org/licenses/by-nc-nd/3.0/)

* Corresponding author.

E-mail address: didier.raoult@gmail.com (D Raoult).

ContentslistsavailableatScienceDirect

j o urn a l hom e pa ge : ww w e l s e v i e r c om/ l o ca t e / i j i d

http://dx.doi.org/10.1016/j.ijid.2014.07.021

1201-9712/ß 2014 The Authors Published by Elsevier Ltd on behalf of International Society for Infectious Diseases This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/3.0/ ).

withthis systemand foundthreemitestobepositive:thetwo

alreadyidentifiedby16SrDNAandanotherfromthesamepatient

B.quintanawasfinallyidentifiedinthreeoutofthesixDemodex

mitescollectedfromthatpatient

The patient was a 63-year-old woman from Schenefeld/

Hamburg(Germany)whopresentedwith

erythematotelangiecta-ticrosaceadefinedbyamildtelangiectasiaandmoderateflushing,

but neither non-transient erythema nor papules and pustules

Toourknowledge,shewasnothomeless,infestedwithlice,orin

contactwithlice-infestedhomelesspeople.Asnobloodsamples

werecollectedduringthisstudy,wecouldnotperformB.quintana

serology testing to establish whether or not the patient was

contaminatedwithB.quintana.Finally,inthisstudyweidentified

B.quintanain3/72testedmitesin1/13patients

4 Discussion

To date, the louse Pediculus humanus humanus is the only

confirmedvectorofBartonellaquintana,eventhoughthisspecies

hasbeenidentifiedbymolecularbiologyorcultureinmanyother

arthropods, including the house dust mites Dermatophagoides

farinaeandDermatophagoidespteronyssinus3andhaematophagous

mitesofDermanyssussp.2Thisisthefirstreportofthedetectionof

B.quintana16SrDNA sequencesfromDemodexmitescollected

fromapatientwitherythematotelangiectaticrosacea.Itisnotyet

knownwhetherDemodexmitesplayarole inthepathogenesis

ofrosacea orwhether theymay serveas a hostfor Bartonella

Demodex mites are not haematophagous and do not excrete faeces,sothe‘classical’(asfor liceandsand flies,forexample) epidemiological scheme of Bartonella transmission does not appeartobeapplicablehere.However,Demodexmitesarehighly contagiousandthetransmissionofBartonella-infectedDemodex mitesfromoneindividualtoanotherispossible.Furtherstudies shouldbeperformedinordertoclarifytheroleofDemodexmites

ashostsandpossiblevectorsofB.quintana

Conflictofinterest:Nonedeclared

References

163–165.

N Murillo et al / International Journal of Infectious Diseases 29 (2014) 176–177 177