Dermatophyte infections in the Lisbon and Tagus valley

We ec ui ed 41 FLD pa ien s and 43 HEC om i e uni e si y hospi al pneumology depa men s in F ance and Swi ze land. L. co ymbi e a p o eins we e ex ac ed om he e e ence s ain BBCM/ IHEM 3809 isola ed om FLD-linked hay. P o eins we e sepa a ed by wo-dimensional elec opho esis and subjec ed o wes e n blo ing, wi h se a om FLD pa ien s (n=7) o con ols (n=9). FLD-speci ic p o eins we e iden i ied by mass spec ome y (LC-MS/MS) and we e p oduced as ecombinan an igens as p e iously desc ibed 1 . The diag- nos ic pe o mance o ELISA es s using he ecombinan an igens was assessed wi h all he se a om FLD pa ien s and con ols. Resul When compa ed wes e n blo memb anes e ealed by FLD and HEC se um, 25 spo s we e conside ed as FLD speci ic. The 25 FLD speci ic spo s we e cu om he gel and analyzed by LC-MS/MS. Six y-nine di e en p o eins we e iden i ied om he 25 spo s. Six p o eins we e selec ed o be p oduced as ecombinan an igens: acylCoAdehyd ogenase, p o easome alpha, py u a e kinase, mala e dehyd ogenase, ATP syn hase alpha, dihyd olipoyl dehyd ogenase. ELISA es s we e pe o med using each ecombinan an igen, wi h all he se a om FLD pa ien s (n=41) and con ols (n=43). Dihy- d olipoyl dehyd ogenase was he mos e ec i e ecombinan an igens o disc imina ing FLD pa ien s om con ols, wi h AUC =0.82 and wi h sensi i i y and speci ici y o 81% and 77%, espec i ely. ELISA using p o easome a also showed AUC abo e 0.80, wi h sensi i i y o 88%, bu sensi i i y was only 65%. Conclusion In ol emen o L. co ymbi e a in FLD has been desc ibed mainly in Eas o F ance and Finland. Combining ecombinan an i- gens om L co ymbi e a wi h ecombinan an igens om o he mic o-o ganisms (Saccha opolyspo a ec i i gula, Aspe gillus) in ol ed in FLD would be p obably help ul o p oduce a s anda dized ELISA ki e ec i e o diagnosing FLD wha e e he geog aphic loca ion o he pa ien . A p ospec i e s udy, using such a es combining mos e ec i e ecombinan an igens om S ec i i gula 1 and om Aspe gil- lus2 wi h dihyd olipoyl dehyd ogenase and p o easome alpha om L. co ymbi e a, is ongoing in ou lab o assess diagnosis pe o mance wi h pa ien s om di e en geog aphic o igins. 1. Ba e a C, Millon L, e al. Immuno eac i e p o eins o Saccha- opolyspo a ec i i gula o a me ’s lung se odiagnosis P o eomics Clin Appl. 2014: 8(11–12):971–81 2. Millon, Roussel e al. Aspe gillus ecombinan an igens o se o- diagnosis o a me ’s lung disease J Alle gy Clin Immunol, 2012, 130(3):803–805. P471 Va ia ion in he polysaccha ide capsule size in e e es wi h iden i ica ion o C yp ococcus neo o mans and C. ga ii by MALDI-TOF mass spec ome y D. Y. Thomaz, 1 M. S. M. Vidal, 2 R. C. G en ell, 3 M. C. Giudice, 2 L. Juliano Ne o, 3 G. Bena d, 4 G. M. B. del Neg o 5 and J. N. Almeida J 5 1 School o Medicine - Uni e si y o S~ ao Paulo, S~ ao Paulo, B azil; 2 Ins i u e o T opical Medicine, Uni e si y o S~ ao Paulo, S~ ao Paulo, B azil; 3 School o Medicine, Fede al Uni e si y o S~ ao Paulo, S~ ao Paulo, B azil; 4 Medical School, Uni e si y o S~ ao Paulo, S~ ao Paulo, B azil and 5 Clinics Hospi al, School o Medicine, Uni e si y o S~ ao Paulo, S~ ao Paulo, B azil Objec i es o in es iga e he in e e ence o C yp ococcus’s capsule size in MALDI-TOF MS species iden i ica ion. Me hods ou C. neo o mans e e ence s ains, WM 148 (VNI), WM 626 (VNII), WM 628 (VNIII), WM 629 (VNIV), and ou C. ga ii, WM 179 (VGI), WM 178 (VGII), WM 161 (VGIII), and WM 779 (VGIV) we e used o capsule g ow h expe imen s. Ini ially he yeas cells o each s ain we e incuba ed o e nigh in acid Sabou aud b o h a 30 °C wi h shaking. A e cen i uga ion, he pelle s we e incuba ed in he capsule g ow h inducing medium (Sabou aud b o h dilu ed 10 imes, pH 7.3) a 37 °C wi h shaking. The capsule induc- ion was ca ied ou eplacing medium e e y 48 h du ing a pe iod o 20 days. To educe he size o he capsules, he yeas cells we e hen seeded in acid Sabou aud aga plus 2.9% NaCl, and incuba ed a 30 °C o 48 h. Fo each s ain, he capsule size was measu ed in ligh mic oscope using he so wa e Axio ision. A leas en di e en ields o he slides we e andomly chosen and 40 o 50 capsule cells we e measu ed. Subsequen ly, he s ains we e analyzed by MALDI- TOF mass spec ome y a e s anda d ex ac ion p o ocol wi h e ha- nol 70% and o mic acid 70%. Then, he supe na an s we e placed in MALDI a ge pla e wells, d ied a oom empe a u e and o e laid wi h he ma ix con aining a sa u a ed solu ion o HCCA. Mass spec- a (MS) we e gene a ed by he Au o lex MALDI-TOF mass spec om- e e and compa ed o main spec a o bo h C. neo o mans and C. ga ii om he da abase Bio ype 3.1. To ensu e he ep oducibili y o spec a each s ain was es ed in quad uplica e in eigh di e en expe imen s. Resul s we e exp essed as log sco e o 1.700–1.999 indica ing a p obable genus iden i ica ion and log sco e abo e 2.000 indica ing secu e genus and species iden i ica ion. Resul s he s ains o C. ga ii de eloped la ge capsule sizes in com- pa ison wi h C. neo o mans s ains (mean 16.8 lm95.6 lm, P=0.002) when incuba ed in he induc ion medium. Capsule size was in e sely ela ed o MS quali y. All eplica es wi h capsule size abo e 10 lm had un eliable species iden i ica ion. The mean capsule size o C yp ococcus s ains wi h co ec species iden i ica ion was 2.18 lm, whe eas o hose assigned wi h log sco e below 2.000 was 5.77 lm (P=0.03). Conclusion The capsule o C yp ococcus nega i ely in e e ed wi h he pe o mance o MALDI-TOF MS species iden i ica ion. Capsule size educ ion is ecommended o achie e a eliable labo a o y iden i ica- ion o C yp ococcus isola es by his echnology. P475 De ma ophy e in ec ions in he Lisbon and Tagus alley C. Ve ı ´ssimo, 1 J. C. B and~ ao, 1 H. L. Sim~ oes 2 and R. F. P. Sabino 1 1 Na ional Ins i u e o Heal h D . Rica do Jo ge, Lisboa, Po ugal and 2 INSA, Lisboa, Po ugal Objec i e A e ospec i e s udy was done in he Po uguese Na ional Ins i u e o Heal h, in o de o es ablish he p e alence and cha ac e ize de ma ophy ic in ec ions wi hin he NUTSII egion o Lisbon and Tagus Valley. Ma e ial and me hods This e ospec i e s udy included 4193 bio- logical samples om pa ien s wi h medical suspicion o ungal in ec- ion, collec ed om 2004 o 2013. Samples we e ob ained by ex ac ing hai s and sc aping skin and nails using a s e ile cu e e o e he a ec ed a eas. Samples we e mic oscopically examined a e 30% W/V o 20 min po assium hyd oxide p epa a ion and cul u ed on Sabou aud dex ose aga wi h cicloheximide. Species we e iden i- ied by he obse a ion o mo phological ea u es ha included col- ony pigmen a ion, ex u e, g ow h a e and dis inc i e mic oscopic s uc u es. Physiological es s (u ea, i amin and amino acid es aga s) we e used whene e necessa y. Resul s The a e age equency o de ma ophy e in ec ions was 21%, anging om 18% o 26%; 841 indi iduals (425 emale and 410 male) had posi i e cul u es. Tinea capi is was con i med in 236 (28%) pa ien s and was mo e p e alen in child en om he g oup o 1– 9 yea s old. In scalp de ma omycosis, Mic ospo um audouinii was he mos equen ly isola ed species (N=120, 51%), ollowed by T. soudanense (N=44, 19%). Males we e mo e a ec ed (58%) han emales (42%). Onychomycosis caused by de ma ophy es we e con i med in 385 cases (46%). O he ungi ecognized as cause o onychomycosis we e no conside ed o his epo . Skin samples we e posi i e in 220 cases (26%). The mos p e alen de ma ophy es isola ed in nail and skin sam- ples we e T. ub um (206 and 93 isola es, espec i ely) and T. men a- g ophy es wi h (49 and 39 isola es espec i ely). E. loccosum was he species less equen ly ound in skin samples (1%). ª2015 The Au ho s Mycoses ª2015 Blackwell Ve lag GmbH, 58 (Suppl. 4), 51–226220 Pos e P esen a ions Conclusion The equency o in ec ions by de ma ophy es has e ealed i sel s eady o he las 10 yea s. The spec um o de ma o- phy es is simila o hose epo ed in o he s udies o inea co po is and onychomycosis, being T. ub um he main ae iological agen . Howe e , ou esul s di e ed om se e al s udies ha desc ibe a aise, du ing he ecen yea s, in he numbe o cases o inea capi is caused by M. canis, especially in o he Medi e anean coun ies. Ou esul s showed a high p e alence o an h opophilic species (M. audouinii and T.soudanense) in he egion o Lisbon and Tagus alley, whe e he numbe o o eign ci izens om A ican coun ies is highe . Like in o he s udies he numbe o in ec ions by E. loccosum was e y low sugges ing ha he e was a eplacemen o his ae iological agen by o he de ma ophy es like T. ub um. P476 In ea men o p ima y cu aneous Aspe gillosis, sys emic an i ungal he apy and su gical managemen ; P esen a ion o wo cases V. A kan-Oguz, 1 I. S. Sa oglu, 1 M. Celik, 1 A. E. Acan 1 and N. Yapa 2 1 Dokuz Eylul Uni e si y Facul y o Medicine, Izmi , Tu key and 2 Dokuz Eylul Uni e si y, Izmi , Tu key Objec i es Aspe gillus skin in ol emen occu s by hema ogenous sp ead o local inocula ion. P ima y cu aneous aspe gillosis ha appea s wi h local inocula ion is a ely de e mined on immunocom- pe en pa ien . Howe e , i is an impo an cause o mo bidi y and mo ali y in su gical pa ien s. T ea men is con o e sial and di e - en medical and/o su gical app oaches can be applied. He e, wo cases o p ima y cu aneous aspe gillosis diagnosed wi h he g ow h o Aspe gillus la us on issue samples a e p esen ed. In he ea men o pa ien s, o iconazole and su gical ea men was applied oge he . Di e en me hods we e used o he wound ca e. Me hods Pa ien 1: 53 yea s old emale, who had a mo o cycle acci- den esul ed wi h le leg inju y. She had a wound wi h a la ge is- sue de ec on he le c u is wi h ibia and ibula ac u e. Toge he wi h ac u e ea men epea ed su gical deb idemen we e applied on he wounds ha had wide issue de ec s. Bu in 3 weeks o su gi- cal ea men due o he appea ance o in ec ed nec o ic issue, he wounds we e deb ided again o he issue cul u es. A. la us has g own in cul u e. Fo he dis inc ion o colonisa ion and in ec ion, issue cul u es we e epea ed o wice. A. la us has also g own in bo h. Fungal spo es and hyphae s uc u e b anching wi h na ow angle we e demons a ed. Because o ha ing nec o ic a eas wi h he con inuing in ec ed appea ance and g owing o A. la us in 3 conse- quen issue cul u es she was ea ed wi h ecu en su gical deb ide- men and i o iconazole. An ibac e ial abso ben polyme d essing - So bac abso p ion d essing; SB was also used o he wound ca e. A e hi d week o he ea men he e was no ungus g ow h in cul- u es. Recons uc ion was made wi h he skin g a aken om la - e al side o igh high. Vo iconazole he apy was comple ed in 12 weeks (Figu e 1). Pa ien 2: 39-yea s old male pa ien applied o eme gency se ice wi h bu n and ca bon monoxide in oxica ion. Because o he de elop- men o igh o ea m, igh high and c u is compa men synd ome, ascio omies we e pe o med o bo h ex emi ies. He s ayed in ICU o 16 days. In ec ed nec o ic wounds we e deb ided o wice. A. la us has g own in he issue cul u e. He was ea ed wi h o iconazole and igecycline. Vacuum-assis ed closu e (VAC) he apy was used o wound ca e. The e was no any g ow h in he issue cul u es aken in he second week o he an i ungal ea men . Following nega i i y in he issue cul u es, wound was p ima ily su u ed. He was ea ed success ully wi h o iconazole o 8 weeks (Figu e 2). Discussion P ima y cu aneous aspe gillosis may de elop a in a- enous ca he e inse ion si es and unde adhesi e ape d essings and bandages o in cases whose skin in eg i y is impa ed. Bo h o he epo ed cases a e p ima y cu aneous aspe gillosis. I has been hough ha slow p og essi e massi e nec osis migh be guiding o he oppo unis ic ungus in ec ion in bo h cases. In ea men o cases, epea ed deb idemen / di e en wound ca e and sys emic o iconazole ea men was adminis e ed oge he . In slow bu p o- g essi ely des uc i e pos - auma ic wound in ec ions, p ima y cu a- neous aspe gillosis should be conside ed in di e en ial diagnosis. P477 Fusa ium solani isola ed om pa ien s wi h onychomycosis: nail in ec ion po en ial and bio ilm o ma ion abili y J. Galle i, 1 F. K. Tobaldini, 2 S. Sil a, 3 E. S. Kioshima, 4 M. Neg i 4 and T. I. E. S idzinski 4 1 Uni e sidade Es adual de Ma inga, Ma ing a, B azil; 2 Uni e sidade Es adual de Ma ing a/ Uni e sidade do Minho, Ma ing a, B azil; 3 Uni e si y o Minho, B aga, Po ugal and 4 Uni e sidade Es adual de Ma ing a, Ma ing a, B azil Objec i e The aim o he p esen s udy was o pe o m an epidemio- logical s udy, e alua e he abili y o Fusa ium solani o use he human nail as a single sou ce o nu ien s and i s po en ial o bio- ilm o ma ion. Me hods We i s pe o med an epidemiological s udy o de e mine he equency o F. solani in pa ien s wi h onychomycosis. The s udy included da a om all pa ien s who a ended he Teaching and Resea ch Labo a o y o Clinical Analysis (LEPAC), Di ision o Mycology, Uni e sidade Es adual de Ma ing a (UEM), be ween Jan- ua y and Decembe 2013 wi h suspec ed onychomycosis. This desc ip i e, e ospec i e, c oss-sec ional, obse a ional s udy was ª2015 The Au ho s Mycoses ª2015 Blackwell Ve lag GmbH, 58 (Suppl. 4), 51–226 221 Pos e P esen a ions