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Giovanna SIMONETTI * Correspondingauthor: Key head Running 4 Descartes, Paris,France. 3 Italy 2 Tunis ElManar 1 Najla Sadfi Touka Nourchéne foot molds in mycosis involved I This article isprotected rights by All copyright. reserved. 10.1111/myc differencesofversion Record. and c theVersion Please between this throughcopyediting,typesetting, the proofreading pagination and process, whichmay lead to This article undergone has for and buthas accepted been been not review publication fullpeer i VD rA I e Dy ) 1434 Biomarkers and Pathologies Mycology, of Laboratory vitro n Department ofDermatology and Venereology Department of Public Health and Infectious Diseases, Infectious and Health Public of Department AcceptedDepartment Article
- words 1034
- biofilms biofilms 0000 : -
Tinea pedis Zouaoui .1 : Dy I e yt e e yt Ie Dy - 2706 : 0000 f Microbiology, of 2092 Tunis, Tunisia 2092 Tunis, foot bri : n atfna susceptibility antifungal and VDI d R R d VDI mycosi 1 1
and GiovannaSimonetti , Serena Corpologno Serena ,
,
onychomycosis, dermatophytes s: b ( ITRVO OO R NRAVOI iofilm iofilm
ôia Necker Hôpital
and drug sensitivity 2 , Marie , , Hôp 2 *
- E f emtpye n non and dermatophyte of ital Laital Tunis,Tunisia. Rabta, fns aae AP Malades nfants - Elisabeth Bougnoux Elisabeth , Faculty , “
fungal biofilm, susceptibility biofilm, fungal tests. Sapienza
of ” University of Rome of University ” Sciences of Sciences ite t his article ashis doi: provided byArchiviodellaricerca-UniversitàdiRomaLaSapienza - P University HP, 3 , Dalenda El Euch El Dalenda ,
Tunis,
-
dermatophyte University , Rome, Rome, ,
Paris
brought toyouby
4 - , CORE
like species other onychomycosis, including infections, foot to associated commonly most species Aspergillus are rubrum Trichophyton affect in diseases fungal commonest the among are onychomycosis and pedis Tinea Introduction appropriate mycosis foot therapy of formation biofilm fungal with obtained growth biofilms staining b fungal against drugs of broth CLSI using out tested isolated mycosis foot in involved the resistance drug mycosis foot D world. Background Summary E Ph Italy.Aldo Moro5,00185,Rome, Health Public of Department T - his article isprotected rights by All copyright. reserved. mail: mail: one/Fax number: 064468626 (+39)
Accepted Article susceptibility A. candidus, A. fumigatus, A fumigatus, A. candidus, A. also isolated from diseased nails such as as such nails diseased from isolated also
MC of MICs . the elderly, children and adults and children elderly, the
with MIC values MICvalues with
. [email protected] from in vitro in raohts n, l and, ermatophytes Results:
sp : and
Tunisian t p Tinea pedis and onychomycosis are among the commonest fungal diseases in the the in diseases fungal commonest the among are onychomycosis and pedis Tinea erbinafine
. Objectives:
flu after 4
- ol be could to 6 ims qatfcto rvae ta al pce studied species all that revealed quantification Biomass . conazole, conazole,
Among the known known the Among
niugl drugs antifungal microdilution method microdilution is 72h. 72h. patients with foot mycose foot with patients
iofilm the most common specie common most the ranging from 0. fromranging
against . .
F Methods: be o om biofilms form to able Conclusion: This work aims to aims work This lu
s frequently, ess and Infectious Diseases “Sapienza”, University of Rome, Piazzale Rome, of University “Sapienza”, Diseases Infectious and econazole
conazole, econazole, itraconazole and terbinafine inhibited inhibited terbinafine and itraconazole econazole, conazole, formation
. Fusarium Fusarium
niger .
/ (+39) 064468625 1
-
3 T of . carried were griseofulvin and terbinafine itraconazole, , , A. flavus, A. terreus, A. terreus, A. flavus, A. , 031 to >64 to 031
hirty The most The Aspergillus has been has
These findings can help clinicians to develop the the develop to clinicians help can findings These sessile sessile . solani
The ability to form biofilm and a and biofilm form to ability The non - six six evaluate - . s, and s, emtpye mo dermatophyte
.
but various non various but emtpye and dermatophytes emtpye and dermatophytes
E quantified by Crystal by quantified frequently identified fungi are fungi identified frequently
cplross brevicaulis Scopulariopsis µg ml µg Fungal conazole showed the highest the showed conazole
spp.,
identified with MALDI with identified in vitro in - 1 . A. ifl has biofilm The best antifungal activity has been activityhas antifungal best The
v ersicolor
the ability to form biofilm and biofilm form to ability the ochraceus and ochraceus - dermatophyte lds non non are eosrtd increasing demonstrated
Violet and Violet and and
- - etiological agents of of agents etiological dermatophyte dermatophyte were . sydowii A. ntifungal activities ntifungal ,
- the world; the
Fusarium
TOF A. sclerotiorum A. molds (NDMs) molds dermatophytes. activity against activity
able to form form to able Safranin Red Red Safranin have been have
are the the are fungal
molds molds
these sp p .,
hyopru keratinophylum Chrysosporium including species Tunisia in enrolled d the of Unit Mycology the attendingmycosis foot with patients from isolated were strains Clinical Fungal methodsMaterials and Violet and Safranin ofbiofilmstainingamount quantified Red the ofsusceptibility to were study present the of aims The metabolicallyactiveinactive and cells in 19 agent antimicrobial 16 Derm necessary efficacy commercially the to resistance of emergence study limitation Furthermore, long term, several with cases ofrecurrence loca and type the to treatment The term attendi cases of T onychomycosis incriminated also are T er hese infections are infections hese his article isprotected rights by All copyright. reserved. , - 20 Accepted18 Article matology . . therapyand h Citl violet Cristal The Fungal
atophytes and atophytes f niugl ucpiiiy ehns constitutes mechanism susceptibility antifungal of
including including strains of an antifungal antifungal an of
s Scopulariopsis Scopulariopsis 15
with high toxicity, toxicity, high with .
biofilm D
of foot mycosis foot of systemic
epartment ng the dermatology dermatology the ng caused clinical isolates of isolates clinical difficult eradicationdifficult recurrentchronic of nails T
s richophyton tion of lesions of tion 22 NDMs
18 considered as an important public health public important an as considered represent an important an represent .
. 7 Thirty six strains were included were strains six Thirty by Many methods Many , 8 assay . niugl agents antifungal brevicaulis n the in Fusarium F
drug, have the sru solani usarium
can be local be can a be dmntae t be to demonstrated been has ay drug many
rubrum n2 and (n=2) University o ht h dvlpet f e ad oeta cmons is compounds potential and new of development the that so . dermatophytes dermatophytes departments and departments Currently
ability species
( evaluate n=3), have been developed to evaluate the quantity of quantity the evaluate to developed been have
(n=21) mat
s 13 but essential systemic treatment is required is treatment systemic essential but to
seglu terreus Aspergillus Fusarium solani Fusarium 9 and optl a at (Tunis) Rabta la Hospital present ure biofilms . role interaction . adhere the
the the , vial aet ad m and agents available Trichophyton F in the pathogenesis and pathogenesis the in treatment of treatment and usarium usarium in vitro in
this can this on many disadvantages disadvantages many NDMs
s
in this st this in biotic or abiotic or biotic
and and 21
ability to form biofilms and biofilms form to ability . n motn strategy important an
oxysporum
be h ms reliable most the (n=2 resistance
implicated in foot mycosis foot in implicated dermatophyte infections dermatophyte problem, constituting a large bulk large a constituting problem, interdigitale infection
due (n=1). udy .
), Fusarium oxysporum Fusarium ),
to the high prevalence, high the to
(Table
ni a until
T. T. r eti are 10 surface formingsurface biofilms 14 ay help to provide the the provide to help ay - in the in 12 . mentagrophytes
. n nte hn, the hand, another In such (n=5),
1)
lgcl agents ological , prosp test 26
resistance to the to resistance
a , o restrict to and 10 10 and s dermatophyte
hc stains which ective therapeutic
depending antifungal is usually is .
biofilm Crystal
(n=2 molds molds DSM study long
the the of of ), -
microscopy. 0.9% 24 to with supplemented medium 28° agar dextrose Potato on grown were was assay biofilm The of biofilmEvaluation the lowest 100% thatinhibited ofgrowth. drug concentration MICinhibition; 0.032 to 16 ( USA) agents: antifungal five 0.4×10 of concentration 28 at USA) Missouri, fungi filamentous a method microdilution (MIC) concentration inhibitory minimal The susceptibilityAntifungal assay LouisAldrich, St. USA) Missouri, AP Hospital microscopic MALDI using ( and protein of analysis by macroscopic confirmed was strains on fungal of identification based methods standard using Germany) Braunschweig, and 4870 T Parasitology ITC his article isprotected rights by All copyright. reserved. Accepted Article C ) -
, , well plates well ni sporulation until
o rmvl f non of removal for t wit erbinafine cnetain rne fo 64 from ranged concentrations h . terreus A.
µg ml µg - - P Prs France Paris, HP, yooy nt n h Dprmn o Mcoilg, Necker Microbiology, of Department the in Unit Mycology 80 .
After After was the lowest concentration that caused 80% growth inhibition andgrowth theMIC caused80% inhibition concentration that lowestwas the ( 23 - TRB 1 . .
All strains were grown on Potato Dextrose A Dextrose Potato on grown were strains All MIC -
30°C
. formation flu ccording to the Clinical and Laboratory Standards Institute reference Institute Standards Laboratory and Clinical the to ccording 24 performed w performed 4 ) S 195 DSM h iouu ws tnadzd o 1×10 to standardized was inoculum The
to 5×10 to and , conazole 50 -
72h at 37°C at 72h were
- L was defined as the lowest concentration that caused ≥ 50% growth 50% ≥ caused that concentration lowest the as defined was until conidia formation. conidia until adherent cells. Morphology of biofilm biofilm of Morphology cells. adherent g - glutamine, buffered with MOPS acid MOPS with buffered glutamine, riseofulvin ).
sda rfrne strains references as used 4
slts ee trd at stored were Isolates
CFU with 30% glycerol of the time with 30%until use. ( 8 FLC ith the use of methods described methods of use the ith
rm emn olcin f irognss (DSMZ, Microorganisms of Collection German from Sga lrc, t Lus isui USA) Missouri, Louis St. Aldrich, (Sigma , the cells were washed two times with sterile saline sterile with times two washed were cells the ,
ml ) ,
(GSF) - e 1 conazole . The .
o .2 µg 0.125 to on planktonic cells planktonic on
(Sigma Aldrich, St. Louis Missouri, USA) Missouri, Louis St. Aldrich, (Sigma in vitro in
(ECO final at prepared was suspension Inoculum
antifungal ml . – )
Initially, all isolates were identified identified were isolates all Initially, (Sigma Aldrich, St. Louis Missouri, Missouri, Louis St. Aldrich, (Sigma
- 0C n aoru Broth Sabouraud on 80°C 1
resp was determined using the broth the using determined was 6 gar (Sigma Aldrich, St. Aldrich, (Sigma gar
ciey n for and ectively conidia (Sigma activity previously was
characteristics. Then, Then, characteristics.
–
ml was evaluated was Aldrich), observed by light light by observed - - Enfants Malades Malades Enfants 1
19 n PI 1640 RPMI in , , 24 nuae at incubated i . traconazole
A and - TOF MS MS TOF ll strains ll
(Sigma
added added Louis water using using from for for 100
biofilm data The activities antifungal The Statistical analysis ofbiof 50% inhibition described previously as staining violet crystal using quantified µg 1 and 2 4, 8, (16, TER and ECZ of concentrations w and Aldrich) L with supplemented medium 1640 RPMI standardized was inoculum USA), The Missouri, sporulation. Louis until St. 28°C Aldrich, at (Sigma incubated agar dextrose Potato on grown were strains All susceptibilityAntifungal of densitywas nm. OD read at 492 washed 50 with stained was well Each min. 30 for 50°C of 200µl 96 in 72h for formation biofilm the After S reader the Finally, min). transfer 1 was (~ well each dissolved from completely solution was violet crystal the of rest the until pipette a with homogenized gently was solution This well. each to solutions ethanol/acetone 80:20 of 1ml de were biofilm and violet crystal of excess remove to water sterile Crystal V A Cristal violet staining stain and quantify comprising total biomass Totaland Biomass EPS fungi Crystal Biofilm quantification T afranin staining his article isprotected rights by All copyright. reserved. Accepted Articleand min 10 for temperature room at dried are plates the formation, biofilm fter at 570nm.
Violet content and, the MIC the and, content he tms ih 0 µ o PS ni te uentn sae cer Fnly the Finally, clear. stayed supernatant the until PBS of µl 200 with times three phosphate iolet solution was added to each well for 30 min. Thewashed30 min. wells werewith each twotimes well added for was to iolet solution f niugl ciiy (MIC) activity antifungal of
19 as allowed to form biofilm in 96 in biofilm form to allowed as , 24
All experiments were performed intriplicate.All experimentswere performed
and Safranin Red Safranin and
buffered saline (PBS) (Sigma Aldrich, St. Louis Missouri, USA) Missouri, Louis St. Aldrich, (Sigma (PBS) saline buffered ilmtreatment formation duetodrug
are
the dermatophyte
values of of values
result of three independent experiments performed in performed experiments independent three of result 25 e t a e 96 new a to red
bind to negatively charged molecules and can be used to used be can and molecules charged negatively to bind ae en presented been have the - well plates, the plates were washed three times with times three washed were plates the plates, well
- different different and and ltmn ad ufrd ih OS cd (Sigma acid MOPS with buffered and glutamine -
NDM well polystyrene plates in the presence of f of presence the in plates polystyrene well µl of safranin solution 1% for 5 min, min, 5 for 1% solution safranin of µl strains -
el lt ad hn ed n a in read then and plate well
biofilms biofilms ml
have - 1 ) Pearson’s correlation coefficient correlation Pearson’s . After incubation After .
s median. as
been determinated been 20
, 23 .
colorized by the addition of addition the by colorized T he concentrations causing concentrations he
o 1×10 to
n re t rlt the relate to order In , the , 6 conidia .
1ml
biofilm was was biofilm and dried at dried and microplate microplate
duplicate. duplicate.
and then and
of 0.5% of
ml optical optical - 1
ive (r) (r) in –
assay showed that at concentration of 4µg ml 4µg the of concentration at that inhibition; showed assay of percentage the of terms in measured was formation biofilm against and inhibition of percentage of activity antifungal The ITC FLC tested wasThe amount ofmature biofilm i correlation between violet Crystal all of quantification biomass of results The polystyrene surface. brevicaulis 1). (Fig. biomass of amount high biofilms, M12) and M6 biofilms, diffe however biofilm form to ability the demonstrated strains the All ml Trichophyton spp the All (MIC tested antifungal (MIC Fusarium oxysporum against The Results a correlation less than and positively estimated been have T his article isprotected rights by All copyright. reserved.
Accepted - Article 1 - . results of antifungal activity antifungal of results
MIC 50 Trichophyton >16 planktonic cells planktonic 80
T.interdigitale spp ,
-
values ( (M3, M6) resistant clinical isolates, showed the most capacity to form biofilm on a on biofilm form to capacity most the showed isolates, clinical resistant M6) (M3, MIC µg . terreus A. .
rences were observed among them them among observed were rences , linearly related. The zero The related. linearly
ml S. 80 ,
- causing 1 values ( values brevicaulis r= ) and GSF (MIC GSF and )
Te orlto cefcet ls to close coefficient correlation The . spp
0. 0.5 isweak. 0.5 , 100
3534
of dermatophytes ( dermatophytes of C. keratinophylum C. S. (T1) biofilm, biofilm, (T1) . >16
strains were susceptible to susceptible were strains tinea pedis r = r ECO brevicaulis ) and
the results are shown in shown are results the µg 0. and
4422
and (MIC
ml
TRB (Fig. 1 pre the In not A - 80 1 . ), for TRB
showed resi
>16 F. oxysporum F. 50 correlated to correlated
terreus - FLC and MIC
, MIC , value indicates weak relationship weak indicates value a) ITC
µg and Safranin T. rubrum T. gis boim omto ws esrd n em of terms in measured was formation biofilm against (T37) biofilm and biofilm (T37) sent study, the species of species the study, sent - . keratinophylum C. 80 MIC , -
1
80 TER ml values ( ee eitn t FLC to resistant were , the percentage ranged from 0% to 95% for ECO ECO for 95% to 0% from ranged percentage the , , solates were presented in in presented were solates - and MIC and 100 s 1 ) ta , GSF and MIC and GSF , . antifungal activity against all planktonic cellsantifungal activityagainst
n The strains of strains The ( Fg 1 (Fig. TRB (M1 and M2) biofilms, biofilms, M2) and (M1 , r= ce toFLC r= T. T. Figure staining 0. 0. on interdigitale
100 1317) 0584 with MIC with ).
24 )
1 of 2 terreus A. T. T. -
),
. well with MIC . ae en eotd in reported been have are variables the that indicates
amount FLC The effect of antifungal agents antifungal of effect The ECO rubrum 50 F. 80 >64 microtitration plate surface plate microtitration
, ) and and ) (MIC F. solani F.
solani - value ECO MIC
( (M5) biofilm produce a produce biofilm (M5) µg
r = r
80 between the variables, variables, the between T1 T0 n T25) and T40 (T21;
50 , NDMs
80 s iue 1 Figure values of
ml
were resistant to all to resistant were ITC 0. S
of < 0. < of >64 values . 694)
- (M7, M8) and M8) (M7, 1 brevicaulis for FLC for ,
TRB
µg ( (Fig 1 F. 032
show ,
(
r= ≥ 64
ml
solani, F. solani,
and al 2 Table 0.2135) µg , - n vitro in b) 1 ECO , ITC ), µg
. (M3; d a ed ml GSF
- S. S. 1 ). , . . ,
MIC v MIC limited a demonstrate treatment of period short P antifungal agains value MIC highest this In reasonthis important that therapy is it by be preceded drug. sensitivity tests hepatotoxicity, headache, impotency as and rash such skin vomiting), effects diarrhea, (nausea, side disturbance gastrointestinal adverse some with associated been also had a systemic many Although, degradation. biofilm for especially antifungal an use onychomycosis. and and susceptibilitydermatophytes among species. trends resistance the detect to agent antifungal determined T resistance drug of development the also and infection of recurrence rapid the facilitates could inappropriate antifungal toinadequate of selection agents doseduration addition of in and ther represents mycosis diagnosis rates low in 26 are mycosis foot of agents commonest The Discussion susceptibility for However, TRB on thestrains tested. best TheTRB. for 93% to 4% fromand T revious studies revious e ehd o the of methods he his article isprotected rights by All copyright. reserved. , Accepted Article27
also T
RB alue study, study,
To NDMSs like NDMSs
in order to provide the appropriate antifungal agent. agent. antifungal appropriate the provide to order in against s
are the most widely available antifungal agents used for systemic treatment of of treatment systemic for used agents antifungal available widely most the are
our knowledge our 22 >16 low MICs of MICs low to the tested antifungal agents.to the tested , 28 Fusarium -
30
In order to have successful therapy for biofilm onychomycosis, it is necessary to necessary is it onychomycosis, biofilm for therapy successful have to order In 38 µg ml µg . - T.rubrum T.rubrum
41 T a major a
he first step of step first he reported that reported
n vitro in
Fusarium pcrm ciiy to activity spectrum - 1
42
t all the clinical dermatophyte clinical the all t agai pce (2 n M) te n ir boim asy hwd low a showed assay biofilms vitro in the M8), and (M2 species
, fewstudies , ECO .
challenge, frequent challenge, GSF and nst some resistant dermatophyte resistant some nst
antifungal ,
spp., T.interdigitale ITC was the first systemic treatment for skin and nail infections but infections nail and skin for treatment systemic first the was TRB treatment , S TRB
have been been have . results were obtained with the ECO when compared with compared when ECO the with obtained were results has a higher clinical cure with a slower relapse rates relapse slower a with cure clinical higher a has
NDMs brevicaulis
activity
dermatophytes and
dermatophytes of tinea pedis and tinea unguium is to is unguium tinea and pedis tinea of
failures and recurrent infection are observed are infection recurrent and failures
responsible for foot mycosis foot for responsible species causin species GSF
conducted a b ueu t peit h cpct o a of capacity the predict to useful be can
and
have
strains strains Aspergillus
and to eradicate the determined fungal determined the eradicate to and
been reported. H reported. been such as the anthropophilic the as such 43
in Tunisia in , 44
g onychomycosis and tinea pedis. tinea and onychomycosis g isolates 36 .
, oevr GSF Moreover,
37 eety t Recently, .
TRB sp.
44 focused on the antifungalthe on focused can be incriminated be can ,
45 was the most effective effective most the was
owever . I n addition, we note we addition, n Actually, he therapy of foot foot of therapy ntifungal drugs ntifungal demonstrate ,
make precise precise make FLC 34 , T. 35 ITC
rubru n for and had ,
FLC 31
in a in apy apy but the - 33
m a . ,
con fr niugl eitne n onychomycosis in resistance antifungal could for account injury, to resistant surprisingly barrier permeability to contribute a is that biofilm, of production surface. polystyrene a on biofilm form to capacity of and not molds by production biofilm fungi filamentous some of formation biofilm the reported vitro mycosis. foot to associated NDMs and dermatophytes infection cause to ability The and explain be can this tested,antifungal brevicaulis brevicaulis a drugs antifungal systemic to infections fungal these of response therapeutic NDMstinea caused unguium by keratinophylum present the In among species. explain be can (>64 with pedis tinea of treatment oral successful a than pedis tinea in effective against azoles, derivative the Concerning beyond that T zoles his article isprotected rights by All copyright. reserved.
Acceptedsolani F. Article been represent form biofilm in different degrees depending on the species. the on depending degrees different in biofilm form µg ml µg
GSF aaiy f emtpye n NDM and dermatophyte of capacity biofilm forming abilities of abilities forming biofilm
( Trichophyton FLC FLC
described s es active less is -
1 and and
) . ,
(M7, M8) and and M8) (M7,
s their survival, act as a persistent source of infection and further dissemination and dissemination further and infection of source persistent a as act survival, their especially ECO
ed in vivo study, also also study,
F Fusarium Fusarium were
. by the fact that derivate azoles target fungal ergosterol in the structure differs differs structure the in ergosterol fungal target azoles derivate that fact the by
. , oxysporum
Overall, n ITC and
spp isolated f isolated and and
for against .
. in vitro agents of tinea pedis and tinea and pedis tinea of agents
spp. NDMs
n h present the In S. brevicaulis S. pedis. tinea to related species ECO
S ) all .
.
and and rom patients with foot mycosis. foot with patients rom In the other hand, the isolates of isolates the hand, other the In However, TRB However, brevicaulis
is T.rubrum the
this finding is confirmed by study of study by confirmed is finding this resistance ITC T uh as such still not still GSF . ed
isolates rubrum
and by the characteristics of characteristics the by
showed a very high MICs values for the strains of of strains the for values MICs high very a showed
(M3, M6) resistant clinical isolates, isolates, clinical resistant M6) (M3, well standardized pointout authors and many and Fusarium ECO and isolates work
to had and most antifungalmost drugs
C . interdigitale T. present ITC
, we have have we , were .
the ability to adhere to the the to adhere to ability the
T. T. keratinophylum
o om ifls is biofilms form to
.
These finding let us suppose that the high high the that suppose us let finding These mentagrophytes H spp, 58 Otherwise, a first work has reported the reported has work first a Otherwise, demonstrated s owever , 59
a low MICs values for the strains of strains the for values MICs low a unguium
25 These . S
. , assessed assessed h lw susceptibility low The 57
brevicaulis
. However, to our knowl our to However, . Fusarium FLC
T hn h ohr antif other the than
In the present study, present the In F difference he treatment of tinea pedis and pedis tinea of treatment he
46 . associated to associated
solani ,
47
to be the most active agents active most the be to
sho 53 h boim production biofilm the 19 ;
- Decro 56 , and many other studies other many and , otherwise, wed a high MIC values MIC high a wed , eeal rltd to related generally .
species to be be to species
n A. and are resistant to all the all to resistant are s
49 in the susceptibility the in ix. ix. polystyrene - 52
showed the most most the showed foot mycosis foot 1995 . We found We .
terreus ITC ungal agents agents ungal of of the species the 48
refractory was more more was Fusarium edge, the the edge,
showing the poor
surface surface and that
has the by by C in in S S . . .
The authors thatthere declared have of interest. conflict isno Interest of Conflict (Universitarie Tu of Research supported been has study This Acknowledgement in order totarget against action of mode exact their susceptibilities antifungal biofilm test to assays t Therefore, topically antibiotics administered biofilm to and test susceptibility biofilm standardized s its reduce and infection the resolve to help could dose adequate with agents antifungal of selection Appropriate compounds.antifungal of screeningrapid for allow could vitro in performed assaysBiofilm cells. should agents antifungal assay, susceptibility the for and formation biofilm bythe first in explained be can onychomycosis of treatment Successful and therate matrix biofilm of thedrug the in especially biofilm of cycle life the anti in differences The of 2012). pathogenesis the in role of increased which biofilm, important form to an ability and plate nail the represent in dermatophytes of adherence firm biofilm including onychomycosis that suggest factors Many different toantifungal susceptibility drugs T al M8) and (M2 biofilm T he his article isprotected rights by All copyright. reserved. Accepted Article.
25 auain f ifl ad h hg cl dniy n h boim arx a influence may matrix biofilm the in density cell high the and biofilm of maturation .
e nrae lvl o boim eitne neln te motne f developing of importance the underline resistance biofilm of levels increased he C26A15MLP9). ra. n h rcmedtos ie b ECI tee s h ugn ne to need urgent the is there ESCMID by given recommendations the In pread. sessile sessile ii (LR16ES05) nisia
to the tested antifungal agents antifungal tested the to - bi cells. ofilm assays ofilm
virulence and resistance to the antimicrobial agents (Nusbaum et al., al., et (Nusbaum agents antimicrobial the to resistance and virulence
by funds from the Ministry of Higher Education and Scientific Scientific and Education Higher of Ministry the from funds by 60
dermatophyte and NDM and dermatophyte .
,
and from from and diffusion through the biofilm. diffusion through the among dermatophyte and NDM and dermatophyte among .
maturation stage maturation S . Spez” nvriy f oe (Ricerche Rome of University “Sapienza” e etd mn boim ad o planktonic not and biofilms among tested be , c ftr research future uch could - pcfc rapit fr ytmc and systemic for breakpoints specific
confirmed the hypothesis of hypothesis the confirmed
associated to the composition of the of composition the to associated biofilms
are needed to be developed be to needed are
species n niugl drugs antifungal in ,
can be related to related be can
Seidler et Seidler
and and the the
Bacteria, and Cancer Fungi,Malaria, Mechanisms inDermatophytes.In:G, SajidA,Kalia VC,eds. Arora 13. finding inonychomycosis. ofterbinafine study 12. Dermatol. 11. Acad. Dermatol. Venereol. 10. Microbiol. Rev. 9. Aspergillo 8. 2005;373:35 2005/05/01/ 7. Mycoses. yearsfeatures central ofonychomycosisin a Tunisia: 22 retrospectiv 6. onychomycosis. 5. management. 4. Microbiol. andrisk factors tinea p unguium of tinea 3. Am. Acad. Dermatol. 2. canadian patients. surveyof15,000 and epidemiology inpatientsvisiting onychomycosis physicians' offices: of amulticenter 1. References T his article isprotected rights by All copyright. reserved.
Accepted Article Contet I,Dhib ZemniFathallah A, A,Yaacoub Said GahaMB.R, R, Clinical andmycological IA,Farwa Mirza et SA, U,Abbasi al. KaurBhalla P.Onychomycosis Kashyap B, R, GarauPerea A,Noriega Ramos MJ, M,Gonzalez AR, del S, Palacio A.Prevalenceand Gregory onychomycosis populations: patient N.Special inthe HIV LyndeGupta Macdonald Prevalence HC, AK, Cooper CW, Jain EA, P, Summerbell RC. Kumar P, LatkaKumar P, B. C,Taneja Therapyand Antifungal Current Drug Resistance van MB, derArandomi CirkelCrijns Schroeff et PK, al. JG, Elewski toenail BE.effecton patient The onychomycosis quality of oflife. SigurgeirssonB, factors O.Risk Steingrimsson associate Nucci Fusarium infections immunocompromisedpatients. M,AnaissieE.in Negroni Due R.Onychomycosis In: toAspergillus Species.A,ed. Comarú Pasqualotto 2013;56:273 sis: From Preventionsis: Diagnosis to 1997;36:754 2000;38:3226
Indian J. Med.Indian J. Microbiol. - Audonneau N. Les OnyxisAudonneauLes N. ÀMoisissures. 2007;20:695 J. Physicians Surg.Pak. Coll. 1996;35(3 Pt 2):S13 1996;35(3 Pt - 280. - 756. - - 3230. 44. Jan 2004;18(1):48 Jan
-
704.
J.Dermatol. Am. Acad. . Cham: Springer International Springer. Cham: 2017:371 Publishing; 2008;26:108 edis thegeneral in inSpain. population - 16. . Dordrecht:Netherlands; 2010:961 Springer Non -
51. 2011;21(10):597 Br. J. Dermatol.
- de rmatophyte asof moulds pathogens - 116. -- epidemiology, diagnosis and epidemiology, diagnosis and Revue Laboratoires. des Francophone
2000;43:244 - Feb 1992;126 Suppl 39:36 Suppl Feb 1992;126 d with onychomycosis. d withonychomycosis. 600. zed treatment duration treatmentzed e study (1986 e study
Drug ResistanceDrug in - 248. - positive patient.positive
J. Clin. - Int. J. 2007). Clin. Clin. J. Eur. - 971. - - - 385. 39. J.
J. Med. Microbiol. studying theeffectsantifungal multicellular on of fumigatus. communities ofagents Aspergillus 24. 2008;28(16). Approved standard AntifungalReferenceBroth Dilution for Susceptibility Method Filamentous Testing of Fungi ; Ostrosky MMR, Ghannoum C.C. Knapp, M.A., 23. InfectiousMedical Diseases Microbiology. and Etiology,and Factors Risk of Tinea PedisU andTinea 22. human immunodeficiency virus infection. of Candida albicanslevels of unlikelytocontribute is tohigh 21. including complex. parapsilosis strains oftheCandida production and of evaluation antifungalamongst susceptibility clinical Candida 20. 2014;30(6):719 charactevitro 19. 2012;17(7):1 18. 2 2001;291(5505):878 17. Trends Microbiol. 16. for treatment. 15. onychomycosis. 14. T his article isprotected rights by All copyright. reserved.
Accepted Article Jin Y,YipHK, SamaranayakeJin Samaranayake YauBiofilmLP. YH, JY, Melo AS,BizerraFC, Freymuller E, Arthington Costa Nusbaum Charles RS, AG,Kirsner CA.Biofilms indermatology. yeast,ReynoldsBakers' fungal Fink TB, amodel GR. for biof IW.Sutherland matrix The biofilm Pfaller MA.resistance: Antifungal epidemiology, mechanisms, drug and consequ BeutnerAlleyBaker Recent SJ, MRK, KR,Plattnerprogressonthe J. topical therapy of Mowat E, Butcher J, LangMowat E,Butcher J, C, Williams S, ABD,Rex J.H. AndesArthington D., M,SadfiToukabriEuch M,Mokni D,Rouissi N,Dhieb C,El
- - rization of Trichophyton T.mentagrophytes ofrubrumrization and biofilms. 5. Orlandi CB, Sardi JC, FuscoOrlandiSantos CT, JC, CB, Sardi Am. J. Med. J. Med. Am. -
727. Expert Opinion onInvestigational Drugs. Expert May 2001;9(5):222 - Sep 2007;56(Pt 9):1205 Sep 2007;56(Pt Second Edition,M38 Second
- 881. Jan 2012;125(1Suppl):S3 Jan
- 227. - A2. -- J. Microbiol. Clin. -
1212. an immobilized butdynamic environment. microbial ‐ Clinical Standard andLaboratory CLSI. Institute Skaggs B., Brown,B., CV,Espinel Skaggs S.D. ‐ 2017;2017:9. ZeichnerL., M.,Sheeha Pfaller
Ramage G.Developmenta model simple for of - 13. - nguium in Tunisia. inTunisia. nguium Med. Mycol. Almeida AM,Mendes - Skaggs BA, Colombo AL. BiofilmSkaggsBA, AL. Colombo
2007/02/01 2007;16(2):1572007/02/01
Jul 2003;41(7):2961Jul yeastoral incases carriage of Apr 2011;49(3):253 - ilm formation. formation. ilm Zouaoui N.Prevalence, Skin Therapy Therapy Lett. Skin Canadian Journal of Journalof Canadian - Giannini MJ. In MJ. Giannini -
Biofouling. Biofouling. n D.J.,, T.J. W. W. T.J. n D.J.,, forming ability spp. isolates, - ‐ 2967. IngroffA., Science. - 167. -
ences 262.
Jul Jul
Feb
2008;70(6):710 Onychomycosis: AnUpdate. 35. dermatophytosis 34. recurrence. 33. Mar 2010 dermatophytes treatment successful after systemic antifungal with agents. 32. 2013;56(3):327 terbinafine forand dermatophyte nailony abrasion toe 31. 2012;103(6):520 Epidemiologicaletiological andinTunisia. offoot study mycosis 30. onychomycosis. dermatophytes, brevicaulisagents opportunistic Scopulariopsis fungi and other asof 29. 2012;26(12):1479 filamentous fungi from nailinfection. isolated 28. andonychomycosis mycological inGoiania, Brazil. data of 27. prevalence and data r 26. Chemother. reducedantifungal epithelial drug susceptibility cells. on bronchial 25. T his article isprotected rights by All copyright. reserved.
Accepted Article Succi IB,Succi Bernardes I,El FekihBelghith Skhiri N, S, Trabelsi Carrillo El EssalFE,Ataides FS, etyeasts ChaulMH, al.Antifungal of patterns susceptibility and Lemos Silva CR, LK, Costa MR.EpidemiologicalSouza Passos XS, JA, Fernandes OF, M,Piraccini E,BrunoroPapini Difonzo BM, A.EpidemiologyItaly: ofonychomycosis in Salvenmoser S,MullerSeidler FM.fumigatus MJ, Aspergillus fo Shirwaikar T,Shirwaikar Prabhu AA,Lobo KS.Treatment Thomas A, R, of DelGonzalez Palacio A,Garau M, Scher RK,Baran inclinical R.Onychomycosis practice: factorscontributing to LongPiraccini A. A,Tosti BM, Sisti ;62(3):411 Br. J. Dermatol. Br. J. Dermatol. Nov 2008;52(11):4130 - - - Munoz AJ, GiusianoIn et AJ, G, al. GuarroJ, Munoz v 714. 332. . 2000. - Agents.Int. J. Antimicrob. 524. - 1485. -
414. isk factor identification. isk
- Engemann AR, OrofinoEngemann Sep 2003;149 Suppl 65:5Sep 2003;149Suppl Indian J. Pharm.Sci. - 4136.
- Escalada Trends MT. thetreatment A,Calvo in of Aug 2007;30(2):157 - term follow term Mycoses. J. Eur. Acad.J. Dermatol. Venereol. - Aounallah H,Khaled Nov - Costa R. Intermittent R. therapywith Costa - chomycosis: a pilot study. apilot chomycosis: 9. 2015;58:659 - Dec
- up of toenailcaused onychomycosis by itro activityitro against ofvoriconazole Mycoses.
- 161. Actas Dermosifiliogr. Antimicrob. Agents - 664.
Jan 2010;53(1):68 Jan
S, Fazaa B. S, rms biofilms with with rms biofilms
J. Am. Acad. J. Dermatol. Mycoses. Dec - 71. May May
1995;34(2):122 48. mellitus. 47. methods. toantifungalsusceptibilities by agentsazole vitro microdilution and testedin brothmacro 46. against dermatophytes inTokat, Turkey. isolated 45. clinicalIsfahan. isolatesofdermatophytes from 44. Mycopathologia. 43. of thefoot. 42. testing ofTrichophyton spp. 41. Exp.Clin. Dermatol. of fluconazole, voriconazole itraconazole, against and onychomycosis. fungi terbinafine causing 40. Chemother. andantifungaltriazoles established Trichophyton agentsrubrum. against 39. associated with onychomycosis. efinaconazole and currently availableantifungal agents a 38. of dermatophyte causing strains tinea unguium. 37. a casemellitus: strains c 36. T his article isprotected rights by All copyright. reserved.
Accepted Article Bell BarrosA, H ME,SantosDde Bueno Martinez Zapata C, B,GallegoIn activity JG, Sanclemente G,vitro M,Mesa AC. Deng Zhanget S, al.Comparison SeyedmousaviS, the C, of activities invitro of newer Tatsumi Siu WJ, Y,SendaJo al. H,et Sarifakioglu DemirbilekIn vitroantifungal E,Seckin M,Can F. D, suscept InOzcan D, Seckin M. D, Demirbilek antifungal vitro ofdermatophyte susceptibility Decroix J. Tinea pedisDecroix J. (mocassin WS. CommonTan fungalJoseph the feet JS, infections of patients in diabetes with LavradorSiqueira ER,Ferreira Pedroso JC, RS, MA, CandidoRC. InYenisehirliE, Yenisehirli G,Tuncoglu A,Bulutactivities Y. of antifungal vitro drugs ChadeganipourInevaluationgriseofulvin Havaei M,Nilipour S, of vitro A. UpdateGupta AK,inantifungal dermatophytosis. Cooper EA. therapy of ausing and tinea onychomycosis pedis non inpatientswith Drugs Aging. Rev. SaoPaulo. Trop. Inst. Med. The Cochrane The Cochrane - Jul 2015;59(7):4312Jul Syer SE, Khan SM, Torgerson DJ. Oral treatments for Torgerson Oral SM, fungalSyer SE,Khan DJ. ofthe treatments infections skin - - control study. 124. Nov
Aug 2010;35(6):658 - Dec 2008;166(5 2004;21(2):101 database of systematic reviews.database ofsystematic Mycological research. J. Eur. Acad. Dermatol. Venereol.J. Dermatol. Eur.Acad. - Antimicrob. Agents Chemother.Antimicrob. Agents 4314. amdan JS. In methods foramdan antifungal JS. vitro susceptibility - -
6):353 112. - type) treated with itraconazole.type) treated with - 663. Jan
Comparison of in vitro antifungalComparison invitro of of activities - - 367.
Feb 2008; Clin. Exp. Dermatol. Clin. Mycoses. Int. J. Dermatol.
Nov 2006;110(Pt11):1355 gainst a variety of pathogenicgainst a variety fungi of 50(1):1 Dec 2004;47(11 Oct 172012;10:CD003584. - Apr 2013;57(4):1610 Dec 2010;24(12):1442 5.
Dec 2013;52(12):1557 -
insulin Nov 2007;32(6):675 Int. J. Dermatol. Antimicrob. Agents Antimicrob. - - Dermatophyte 12):503 dependent diabetes - 1360. ibility patterns -
507. against
-
1616.
- 1446. - - 1560. Feb 679.
2017;66(7):1045 of and vivobiofilm theinvitro ex 59. Dermatol. 58. 2015;64(11):1436 Candida albicans vitro. Fusarium oxysporumin biofilms and nail 57. 2004;48(1):69 and novel antifungal Fusarium drugs against spp. 56. Antimicrob. Chemother. antifungalprofiles susceptibility ofopportunist 55. complex. haplotypean nomenclature, 54. Microbiol. Infect.Clin. diagnosi 53. infectious agents. tosystemicinsensitive treatmentsrevealsazole terbinafine and non 52. of filamentous fungi. InEstrellamedically M.activity vitro against ofterbinafine important non 51. 1):217 clinical to treatment features and response of59cases. 50. Candida onychomycosis. 49. T his article isprotected rights by All copyright. reserved.
Accepted Article - 224. Al O'Donnell DA,Fothergill K,Sutton A,et al.Molecular phylogenetic multilocus diversity, E,et M,Roilides al.ESCMIDTortorano guidelines and AM,Richardson joint ECMM on Baudraz Garcia A,PiracciniTosti OnychomycosisLorenzi BM,S. caused by nondermatophytic molds: Iorizzo A,PiracciniTosti Lorenzi BM,S, M.Treatment and ofnondermatophyte mold Brilhante RSN, Correia EEM, GuedesBrilhante al. CorreiaQuantitative and RSN, GMdM,et structural analyses Gupta AK,D, Car Daigle Miltefosine QuintanilhaN, S. iseffectiveMachado Rozental Sousa against VilaTV, InOrtoneda JavierI, J, Pastor M,Capilla F, oflicensed Guarro Pujol J. interactions vitro s and management ofhyalohyphomycosis:s and management Fusarium spp.and others. spp.,Scedosporium J. Microbiol. Clin. - 2016;74(6):1241 Hatmi AM,van AD, Diepeningen Curfs
- - Effron G,Effron Gomez 71. - Rosselet F, Ruffieux C, Lurati C, Rosselet F, Ruffieux O,MonodM M,Bontems - 1052. - Dermatology. Dermatology.
1449. J. Antimicrob. Chemother.J. Antimicrob.
Apr 3:27 2014;20Suppl
Apr 2015;70(4):1068 Dermatol. Clin. - d in vitro antifungald invitro resistancetheFusarium solani within species 1246. Aug 2008;46(8):2477 - 2010;220(2):164 viel JL. The role of biofilms in onychomycosis.viel JL. in role The ofbiofilms Lopez A,Rodrigue A,Mellado E,Monzon -
forming ofdermatophytes. ability Jul 2003;21(3):491 - 1071. s in the Fusarium fujikuroi complex. s intheFusariumcomplex. fujikuroi - 46. Jun 2004;53(6):1086 - Diagn. Microbiol. Inf 168. - - 2490. Breuker I,Breuker SpecificHoog GS,Meis deJF.
J. Am. Acad. Dermatol. J. Dermatol. Am. Acad.
- 497, vii. J. Med. Microbiol. - dermatophyte moulds asdermatophyte moulds J. M J. -
1089. ect. Dis. ect. ed. Microbiol. ed. . Onychomycosis - dermatophyte species z
- Tudela JL, CuencaTudela Feb 2000;42(2Pt Jan Jan J. Am. Acad.J. J. Nov -
mean oftriplicates with res of concentrations different using der strains of formation biofilm of percentage Inhibition 2. Figure deviation 492 at absorbance safranin and (A) nm 570 at absorbance Quantification 1. Figure Figure legends treatment 2014. infections ofbiofilm 60. T his article isprotected rights by All copyright. reserved. Table1. Table1.
Accepted Article
Strains
T22 p
T61 T56 T52 T46 T42 T40 T39 T25 T23 T21 T18 T13 T12 T11 T66 T64 T9 T7 T5 T4 HøibyBjarnsholtMoser et C, N, al.ESCMID T,
Identificationdermatophyte of
Sex M M M M M M M M M M M F F F F F F F F F F of biofilm biomass formation after 72h is represented by crystal violet crystal by represented is 72h after formation biomass biofilm of
Clinical diagnosis Clinical
pect control. to Errorb Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis
Tinea pedisTinea pedisTinea pedisTinea pedisTinea Clin. Microbiol. Infect.Clin.
econazole and non a - dermatophyte
rs represent deviati standard
Clinical aspect Clinical and terbinafine. Data are percentage of the the of percentage are Data terbinafine. and
∗
DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO
TDO TDO TDO TDO PSO PSO PSO guideline andfor the diagnosis PH PH PD ID
nm (B). Error bars represent standard represent bars Error (B). nm 2015/05/01/ 2015;21:S1 2015/05/01/ matophyte
clinical strains clinical strains
(MALDI n non and
Identification
on. T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum T.rubrum used study in this
- - dermatophyte TOF MS) - S25.
T T: dystrophic A: onychomycosis. dishydrosis; Plantar hyperkeratosis; Proximal subungual PH: PSO: onychomycosis; TDO: Total NDMs: DLSO:ID: Distal lateral subungualInterdigital; onychomycosis;Plantar PD: his article isprotected rights by All copyright. reserved.
AcceptedTrichophyton Article
T103 T34I M13 M12 T37 T68 T45 T44 M8 M7 M5 M6 M3 M2 M1 T1 .
M M M M F F F F F F F F F F F F
Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis Onychomycosis
Tinea pedisTinea Aspergillus ; C: ; C: Chrysosporium
DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO DLSO
TDO TDO TDO ID ; F: Fusarium
C. keratinophylumC. keratinophylumC.
; S: ; S:
T.interdigitale T.interdigitale T.interdigitale T.interdigitale F. oxysporum F. oxysporum S. brevicaulis S. brevicaulis S.brevicaulis
A. terreus T.rubrum T.rubrum F. solani F. solani Scopulariopsis ;
T40 T39 T25 T23 T22P T21 T18 T13 T12 T11 T9 T7 T5 T4 rubrum Trichophyton (n=26) Dermatophytes Strains Antifungal agents T Table 2. Table his article isprotected rights by All copyright. reserved.
Accepted Article
(n=21) Minimum Minimum i
nhibitory (MIC) concentration of five against antifungal agents MIC 0. FLC 0. 32 16 32 16 64 32 125 8 1 4 4 4 8
5
50
MIC(µg MIC(µg MIC 0. 16 64 16 16 64 64 16 64 32 125 8 4 1 1
80
MIC ml >64 >64 >64 >64 >64 32 64 64 32 16 64 8 2 4 -
1
100 )
<0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 MIC ECO 32
50
MIC(µg <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 MIC 64
80
MIC ml <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 >64 -
1 100 )
<0.031 <0.031 <0.031 <0.031 <0.031 MIC 0 0 0 0 0 0 0 ND ITC ...... 16 . 125 125 031 125 125 062 25
50
MIC(µg <0.031 <0.031 <0.031 <0.031 <0.031 MIC 0 0 0 0 0 0 0 0 . . . 16 . . . 062 125 . 062 . 25 25 25 25 25
80
dermatophyte
ml MIC <0.031 <0.031 <0.031 <0.031 <0.031 0 0 >16 0 0 0 . 0 0 0 062 . - . . . 25 . . . 1
5 5 5 5 5 5 ) 100
<0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 MIC T
and non and R 50 B
MIC(µg <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 MIC -
dermatophyteclinical strains 80
MIC ml <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 -
1 ) 100
MIC GSF >16 ND 0. 0. 0. 0. 0.5 0 0 4 1 1 1 4 . .
5 5 5 5 5 5
50
MIC(µg MIC >16 ND 1 8 2 1 1 2 1 1 8 1 2 1
80
MIC ml >16 >16 ND ND ND 16 2 4 2 2 4 2 2 4 -
1
100 )
M2 M1 oxysporum Fusarium Molds T68 T45 T44 T34I T1 interdigital Trichophyton T103 T66 T64 T61 T56 T52 T46 T42 T his article isprotected rights by All copyright. reserved.
Accepted Article
(n=10) e (n=5) (n=2)
0. >64 >64 >64 >64 >64 0 0. 64 64 16 64 125 4 2 8 .
5 5
0. >64 >64 >64 >64 >64 >64 0 64 16 32 64 125 2 8 2 .
5
>64 >64 >64 >64 >64 >64 >64 >64 >64 >64 >64 64 64 64 1
<0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 < <0.125 <0.125 0.125 16 4 1
<0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 32 8 2
<0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 <0.125 64 16 4
<0.031 <0.031 <0.031 <0.031 0 0 0 0.125 0.065 0 0 0 0 >16 ...... 32 065 125 125 125 062 062 062
<0.031 <0.031 <0.031 <0.031 0 0 0.065 0 0 0 0.25 0 0 > >16 . . . . 065 125 . . 125 . 125 16 25 25 25
<0.031 <0.031 <0.031 <0.031 0 0.125 0 0 >16 >16 . 0 0 0.5 0 0 065 . . . . 25 . . 25
5 5 5 5
<0.031 < <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 0.031 4 1
<0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 8 2
<0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 <0.031 16 4
>16 >16 ND ND ND ND 0 0 0 0.5 0. 1 1 1 1 . . .
5 5 5 5
>16 >16 ND ND ND ND 2 2 1 1 1 2 2 1 1
>16 >16 >16 >16 ND ND ND ND ND 4 4 2 2 4 4
195 A. DSM 4870 T. M13 T37 (n=2) keratinophylum Chrysosporium M5 terreus Aspergillus M12 M6 M3 brevicaulis Scopulariopsis M8 M7 (n=2) solaniFusarium T his article isprotected rights by All copyright. reserved.
mentagrophytes Acceptedterreus Article
8
DSM
(n=3)
>64 >64 >64 >64 >64 >64 >64 64 8 2
>64 >64 >64 >64 >64 >64 >64 64 4 8
>64 >64 >64 >64 >64 >64 >64 >64 16 8
<0.031 <0.031 <0.125 <0.125 <0.125 >64 0 64 16 8 .
5
<0.031 <0.031 <0.125 <0.125 <0.125 >64 >64 >64 16 1
<0.031 <0.031 <0.125 <0.125 <0.125 >64 >64 >64 >64 4
<0.031 <0.031 <0.031 0.065 >16 >16 >64 >16 >16 >16
<0.031 <0.031 <0.031 0.065 >16 >16 >64 >16 >16 >16
<0.031 <0.031 <0.031 0.065 >16 >16 >64 >16 >16 >16
<0.031 <0.031 <0.031 <0.031 <0.031 0 >16 >16 . 1 4 25
<0.031 <0.031 <0.031 <0.031 <0.031 0 >16 >16 . 2 8 25
<0.031 <0.031 <0.031 <0.031 <0.031 >16 >16 16 2 8
0 >16 >16 >16 >16 >16 >16 ND . 1 2 25
>16 >16 >16 >16 >16 >16 ND 0 2 4 .
5
>16 >16 >16 >16 >16 >16 ND 4 1 8
T Acceptedhis article isprotected rights by All copyright. reserved. Article FLC caused ≥ 50% growth inhibition; MICcausedgrowth inhibition; ≥ 50% :
fluconazole; ECO :
econazole;
ITC: TRB: itraconazole; terbinafine; 80
:
the lowestgrowth concentration 80% thatcaused inhibition G S F :
griseofulvin; concentration thatinhibitedg 100% of MIC 50
: the lowest concentration that ;
MIC 100
: the lowest drug rowth.
T his article isprotected rights by All copyright. reserved. Accepted Article (a) (b)