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