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[First case of phaeohyphomycosis due to Pleurostoma ootheca in a kidney transplant recipient in Martinique (French West Indies)]

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Abstract

Phaeohyphomycosis is a group of superficial and deep infections due to dematiaceous fungi. They are most common in tropical environments, especially in immunocompromised hosts. We describe the first case of phaeohyphomycosis due to Pleurostoma ootheca in a kidney transplant recipient in Martinique (French West Indies). A 59-year-old man with a kidney graft, treated with mycophenolate mofetil, tacrolimus, and prednisone, presented suppurative tumefaction of the left ankle. Cutaneous and osseous phaeohyphomycosis caused by P. ootheca was diagnosed, based on mycological, histological, and radiological testing. The patient's condition improved with posaconazole treatment. P. ootheca is a known environmental fungus. Immunocompromised hosts are more vulnerable to many infections, due to opportunistic pathogens. Bacteriological, histological, and mycological testing is required for accurate diagnosis and appropriate treatment. Treatment is not well defined and usually relies on antifungal agents or surgical resection or both. An important point to consider is that azole antifungal agents may cause major drug-drug interactions with immunosuppressive agents such as tracrolimus.
Montrouge, le 17/11/2014
E. Amazan
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Communication courte Me
´decine et Sante
´Tropicales 2014 ; 24 : 323-325
Premier cas de phaeohyphomycose à Pleurostoma
ootheca chez un greffé rénal en Martinique
First case of phaeohyphomycosis due to Pleurostoma ootheca in a kidney
transplant recipient in Martinique (French West Indies)
Amazan E.
1
, Desbois N.
2
, Fidelin G.
1
, Baubion E
´.
1
, Derancourt C.
1
, Thimon S.
1
, Ekindi N.
3
, Quist D.
1
1
Service de dermatologie, CHU de Martinique, BP 632, 97261 Fort-de-France
2
Laboratoire de parasitologie-mycologie, CHU de Martinique, Fort-de-France
3
Laboratoire d’anatomopathologie, CHU de Martinique, Fort-de-France
Article accepte
´le 27/1/2014
Les phaeohyphomycoses sont des mycoses profondes dues
a
`des champignons pigmente
´s du groupe des de
´matie
´s. De
multiples champignons peuvent en e
ˆtre responsables, les
plus fre
´quemment retrouve
´se
´tant Exophiala sp.,Phialophora sp.
et Alternaria sp. [1] Bien qu’ubiquitaires, ces infections
apparaissent particulie
`rement dans les pays tropicaux, le plus
souvent chez des individus immunode
´prime
´s[2].Nous
rapportons la premie
`re observation de phaeohyphomycose a
`
Re
´sume
´.Les phaeohyphomycoses sont des myco-
ses profondes dues a
`des champignons pigmente
´s
du groupe des de
´matie
´s. Ces infections apparais-
sent particulie
`rement en milieu tropical et souvent
dans un contexte d’immunode
´pression. Nous
rapportons la premie
`re observation de phaeohy-
phomycose a
`Pleurostoma ootheca survenue chez
un patient ayant eu une greffe re
´nale en Martinique.
Un homme de 59 ans, aux ante
´ce
´dents de greffe
re
´nale, traite
´par immunosuppresseurs (mycophe
´-
nolate mofe
´til, tacrolimus et prednisone) pre
´sentait
une tume
´faction perce
´e de clapiers purulents,
douloureuse et localise
´e en faces externe et
poste
´rieure de la cheville gauche. Le diagnostic
de phaeohyphomycose cutane
´e et osseuse a
`
Pleurostoma ootheca ae
´te
´pose
´gra
ˆce aux explo-
rations mycologiques, histologiques et radiolo-
giques. Un traitement par posaconazole a permis
une e
´volution favorable. Pleurostoma ootheca est un
champignon saprophyte des ve
´ge
´taux, non de
´crit
comme pathoge
`ne pour l’homme dans la litte
´rature.
Les patients immunode
´prime
´s sont a
`risque de
de
´velopper des infections en tout genre, avec des
germes varie
´s. Des explorations exhaustives (bacte
´-
riologique, mycologique et parasitologique) sont
ne
´cessaires afin d’e
´tablir un diagnostic pre
´coce et
entreprendre un traitement adapte
´. Le traitement
n’est pas consensuel mais empirique, base
´sur des
antifungiques et/ou de la chirurgie. Les antifun-
giques azole
´s posent, dans ce cas, le proble
`me des
interactions me
´dicamenteuses avec le tacrolimus.
Mots cle
´s:phaeohyphomycose, Pleurostoma
ootheca, greffe re
´nale, Martinique.
Correspondance : Amazan E
<emmanuelle.amazan@chu-fortdefrance.fr>
Abstract. Phaeohyphomycosis is a group of super-
ficial and deep infections due to dematiaceous
fungi. They are most common in tropical environ-
ments, especially in immunocompromised hosts.
We describe the first case of phaeohyphomycosis
due to Pleurostoma ootheca in a kidney transplant
recipient in Martinique (French West Indies). A 59-
year-old man with a kidney graft, treated with
mycophenolate mofetil, tacrolimus, and predni-
sone, presented suppurative tumefaction of the left
ankle. Cutaneous and osseous phaeohyphomycosis
caused by P. ootheca was diagnosed, based on
mycological, histological, and radiological testing.
The patient’s condition improved with posacona-
zole treatment. P. ootheca is a known environmen-
tal fungus. Immunocompromised hosts are more
vulnerable to many infections, due to opportunistic
pathogens. Bacteriological, histological, and myco-
logical testing is required for accurate diagnosis
and appropriate treatment. Treatment is not well
defined and usually relies on antifungal agents or
surgical resection or both.
An important point to consider is that azole
antifungal agents may cause major drug-drug
interactions with immunosuppressive agents such
as tracrolimus.
Key words: phaeohyphomycosis, Pleurostoma
ootheca, kidney transplantation, Martinique.
doi: 10.1684/mst.2014.0342
Pour citer cet article : Amazan E, Desbois N, Fidelin G, Baubion E
´, Derancourt C, Thimon S, Ekindi N, Quist D. Premier cas de phaeohyphomycose a
`Pleurostoma ootheca
chez un greffe
´re
´nal en Martinique. Med Sante Trop 2014 ; 24 : 323-325. doi : 10.1684/mst.2014.0342
323
© John Libbey Eurotext, 2014
Pleurostoma ootheca survenue chez un patient ayant eu une
greffe re
´nale en Martinique (Antilles franc¸aises).
Observation
Un homme martiniquais de 59 ans, ayant eu une greffe re
´nale
quatorze ans auparavant et traite
´par mycophe
´nolate mofe
´til
(2 g/j), tacrolimus (3,5 mg/j) et prednisone (10 mg/j), e
´tait
hospitalise
´pour un abce
`s situe
´en regard de la malle
´ole externe
gauche e
´voluant depuis un mois. La le
´sion e
´tait apparue apre
`s
un traumatisme mineur contre un chariot de supermarche
´.Un
traitement par amoxicilline-acide clavulanique avait e
´te
´essaye
´
pendant trois semaines sans efficacite
´.
A
`l’examen clinique, le patient e
´tait apyre
´tique et pre
´sentait
une tume
´faction e
´rythe
´mateuse, perce
´e de clapiers purulents,
localise
´e en face poste
´rieure et externe de la cheville gauche,
douloureuse, avec e
´coulement purulent abondant, sans grains
visibles (figure 1). Les aires ganglionnaires e
´taient libres et le
reste de l’examen clinique sans particularite
´.
La nume
´ration-formule sanguine e
´tait normale et la prote
´ine
C-re
´active e
´tait infe
´rieure a
`10 mg/L. L’examen histopatholo-
gique d’une biopsie cutane
´e montrait un infiltrat inflammatoire
dans le derme avec un granulome inflammatoire e
´pithe
´lioı
¨de et
gigantocellulaire ne
´crosant non case
´eux, et des filaments septe
´s
visibles aux PAS (periodic acid shiff) et au Gomori-Grocott. La
coloration de Ziehl-Neelsen ne mettait pas en e
´vidence de
germes. L’examen direct au May-Gru¨nwald-Giemsa montrait la
pre
´sence de nombreux filaments septe
´s et ramifie
´s. La culture
d’un fragment cutane
´sur milieu de Sabouraud a
`30 et a
`37 8C
isolait des colonies marron a
`beige, en cocarde, finement
duveteuses (figure 2), e
´voluant vers le noir avec le temps,
identifie
´es comme e
´tant Phialophora sp. L’analyse en biologie
mole
´culaire (Centre national de re
´fe
´rence des mycoses et des
antifongiques [CNRMA], Institut Pasteur, Paris) permettait
l’identification de Pleurostoma ootheca.
Le diagnostic de phaeohyphomycose a
`P. ootheca e
´tait
retenu.
Par ailleurs, la culture sur milieu de Lowenstein apre
`s
quatre-vingt-dix jours e
´tait ne
´gative et permettait d’e
´liminer une
infection a
`mycobacte
´rie. Les cultures bacte
´riologiques de pus
et d’un fragment cutane
´restaient ste
´riles. L’IRM de
´celait un
e
´paississement fusiforme du tendon achille
´en et une ne
´crose de
l’os naviculaire (figure 3).
Apre
`sre
´ception de l’antifongigramme, un traitement par
itraconazole (CMI a
`0,750 mg/mL) e
´tait instaure
´a
`300 mg/j, puis
augmente
´a
`400 mg/j. Devant une aggravation des sympto
ˆmes
(augmentation de la douleur, extension de l’inflammation
avec e
´coulement purulent important) a
`un mois et demi de
Figure 1. Tume
´faction e
´rythe
´mateuse purulente de la face externe de cheville
gauche.
Figure 1. Suppurative tumefaction of the left ankle. Figure 2. Biopsie cutane
´e mise en culture sur milieu de Sabouraud.
De
´veloppement de colonies marron a
`beige, en cocarde, finement duveteuses.
Figure 2. The cutaneous biopsy sample was cultured on Sabouraud dextrose agar.
Growth of brown-beige downy colonies.
Figure 3. IRM de la cheville gauche avec e
´paississement fusiforme du tendon
d’Achille et ne
´crose de l’os naviculaire.
Figure 3. MRI of the left ankle: Calcaneal tendon thickening and navicular bone
necrosis.
324 Me
´decine et Sante
´Tropicales, Vol. 24, N83 - juillet-aou
ˆt-septembre 2014
E. AMAZAN, ET AL.
© John Libbey Eurotext, 2014
traitement, l’itraconazole e
´tait relaye
´par du posaconazole
(CMI a
`0,0640) a
`800 mg/j, permettant une e
´volution favorable a
`
deux mois de traitement.
Discussion
Pleurostoma ootheca ae
´te
´identifie
´par Barr en 1985 comme
e
´tant un champignon saprophyte des ve
´ge
´taux en de
´composi-
tion [3]. Il fait partie du groupe des Ascomyce
`tes et de la famille
des Calosphaeriaceae, dans laquelle des cas de phaeohypho-
mycoses ont e
´te
´de
´crits avec Pleurostomophora richardsiae [4]
et plus re
´cemment un cas d’eumyce
´tome a
`Pleurostomophora
ochracea [5].
Ce sont des espe
`ces tre
`s proches morphologiquement de
Phialophora sp., qui ont e
´te
´diffe
´rencie
´es par la biologie
mole
´culaire. Il s’agit de la premie
`re description d’infection
humaine a
`P. ootheca chez un patient ayant eu une greffe
re
´nale.
Les patients ayant subi une transplantation re
´nale sont a
`haut
risque d’infections opportunistes, dont des infections fongiques
superficielles et profondes [6]. L’e
´tude de Revankar a d’ailleurs
mis en e
´vidence que la transplantation d’un organe solide e
´tait
un facteur de risque de phaeohyphomycose [2]. Cependant,
toutes les causes d’immunode
´pression peuvent favoriser les
mycoses sous-cutane
´es et donc les phaeohyphomycoses. En
effet, les phaeohyphomyce
`tes rendent la phagocytose plus
difficile par la pre
´sence de me
´lanine dans leur paroi et
e
´chappent ainsi aux de
´fenses immunitaires [7].
Les phaeohyphomycoses peuvent prendre des pre
´senta-
tions cliniques tre
`s diffe
´rentes d’un patient a
`l’autre (nodule,
kyste, abce
`s...) rendant le diagnostic plus difficile. L’aspect le
plus e
´vocateur e
´tant celui d’un « kyste phaeohyphomycotique ».
Dans le cas pre
´sent, il s’agissait d’une tume
´faction fistulise
´e
pouvant faire e
´voquer une mycose profonde, une infection
bacte
´rienne ou a
`mycobacte
´rie. La ne
´gativite
´des cultures
en bacte
´riologie et mycobacte
´riologie re
´futait ces dernie
`res
hypothe
`ses. L’aspect clinique de la le
´sion et la rapidite
´
d’e
´volution ainsi que l’absence de cellules fumagoı
¨des (corps
scle
´rotiques) et la non-visualisation de grains au sein de la le
´sion
permettaient d’e
´liminer respectivement une chromomycose
ainsi qu’un myce
´tome.
Notre observation rappelle la ne
´cessite
´d’une surveillance
dermatologique rapproche
´e de toute le
´sion cutane
´omuqueuse
persistante ou inhabituelle survenant chez des immunode
´pri-
me
´s afin de re
´aliser les pre
´le
`vements a
`la recherche de
champignons, de bacte
´ries, de mycobacte
´ries, de virus ou de
parasites.
Aucun consensus n’est e
´tabli dans le traitement des
phaeohyphomycoses mais l’itraconazole est l’un des anti-
fongiques les plus efficaces [8]. Le posaconazole a lui aussi
e
´te
´de
´crit comme e
´tant efficient dans les infections fongiques
invasives apre
`s transplantations d’organe y compris les
phaeohyphomycoses [9], comme dans l’e
´tude d’Alexander
(treize patients dont un cas de phaeohyphomycose a
`Exophiala
sp.) [10].
Le traitement des phaeohyphomycoses chez les patients
ayant eu une transplantation d’organe soule
`ve le proble
`me des
interactions me
´dicamenteuses. En effet, les azole
´s sont des forts
inhibiteurs du CYP3A4/5 et peuvent, de ce fait, augmenter les
concentrations sanguines de certains immunosuppresseurs
dont le tacrolimus [11]. Chez notre patient, le dosage du
tacrolimus sous traitement azole
´are
´ve
´le
´un surdosage. Il a e
´te
´
de
´cide
´avec les ne
´phrologues de diminuer le tacrolimus de
3,5 mg/j a
`0,5 mg/j, permettant une normalisation du dosage et
une conservation de la fonction du greffon.
En conclusion, il est important que tous les acteurs
me
´dicaux participant au suivi post-transplantation d’organe
e
´voquent syste
´matiquement le diagnostic de mycoses pro-
fondes – incluant les phaeohyphomycoses –, devant des le
´sions
cutane
´es traı
ˆnantes, afin d’obtenir un diagnostic pre
´coce et
entreprendre rapidement un traitement adapte
´.
Conflits d’inte
´re
ˆt:aucun.
Re
´fe
´rences
1. Ogawa MM, Galante NZ, Godoy P, et al. Treatment of subcutaneous
phaeohyphomycosis and prospective follow-up of 17 kidney transplant
recipients. J Am Acad Dermatol 2009 ; 61 : 977-85.
2. Revankar S, Patterson J, Sutton D, Pullen R, Rinaldi M. Disseminated
phaeohyphomycosis: review of an emerging mycosis. Clin Infect Dis 2002 ;
34 : 467-76.
3. Barr ME. Notes on the Calosphaeriales.Mycologia 1985 ; 77 : 549-65.
4. Ikai K, Tomono H, Watanabe S. Phaeohyphomycosis caused by
Phialophora richardsiae.J Am Acad Dermatol 1988 ; 19 : 478-81.
5. Mhmoud NA, Ahmed SA, Fahal AH, de Hoog GS, Gerrits van den Ende
AH, van de Sande WW. Pleurostomophora ochracea, a novel agent
of human eumycetoma with yellow grains. J Clin Microbiol 2012 ; 50 : 2987-
94.
6. Rubin R. Infectious disease complications of renal transplantation.
Kidney Int 1993 ; 44 : 221-36.
7. Schoeffler A, Redon E, Contet-Audonneau N, et al. Phaehohyphomycose
cutane
´ea
`Cladophialophora bantiana.Ann Dermatol Venereol 2011 ; 138 :
504-7.
8. Sharkey PK, Graybill JR, Rinaldi MG, et al. Itraconazole treatment of
phaeohyphomycosis. J Am Acad Dermatol 1990 ; 23 : 577-86.
9. Scheinfeld N. A review of the new antifungals: posaconazole, micafungin,
and anidulafungin. J Drugs Dermatol 2007 ; 6 : 1249-51.
10. Alexander BD, Perfect JR, Daly JS, et al. Posaconazole as salvage therapy
in patients with invasive fungal infections after solid organ transplant.
Transplantation 2008 ; 86 : 791-6.
11. Vermeire SE, de Jonge H, Lagrou K, Kuypers DR. Cutaneous
phaeohyphomycosis in renal allograft recipients: report of 2 cases and
review of the literature. Diagn Microbiol Infect Dis 2010 ; 68 : 177-80.
Me
´decine et Sante
´Tropicales, Vol. 24, N83 - juillet-aou
ˆt-septembre 2014 325
Phaeohyphomycose a
`Pleurostoma ootheca
... It mimics benign skin and soft tissue tumors such as lipomas, sebaceous cysts or neurofibromas [3,4]. The incidence of phaeohyphomycis is increasing, especially in solid organ transplant patients [5,6]. The most frequently involved genus is Alternaria, but many other species can be isolated (e.g. ...
... The biopsy sample was sent to the mycology laboratory for direct examination, which revealed the presence of numerous septate and branched melanized hyphae with terminal and intercalary chlamydospores (Figs. [4][5][6][7][8]. ...
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We report a case of subcutaneous mycosis in the form of a subcutaneous cyst of the index finger, successfully treated by surgery and posaconazole in an 84-year-old female kidney transplant patient. Intra-operative mycological analysis enabled the diagnosis of Phialophora chinensis phaeohyphomycosis. Phialophora chinensis is an environmental mold recently described in human pathology in cases of chromoblastomycosis. This is the first case of subcutaneous phaeohyphomycosis due to Phialophora chinensis in an immunocompromised patient.
... Infection may result from close contact with Pleurostoma-contaminated plants or direct inoculation of contaminated materials into the body [7][8][9][10][11]. Very often Pleurostoma infections are localized and manifest as cutaneous or subcutaneous lesions and/or nodules [7][8][9][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30], although infections of the bones/joints [9,20,22,[31][32][33][34][35], eyes [10,[35][36][37] and urinary tracts [38] are also observed. In more rare occasions especially in severely immunocompromised individuals, disseminated invasive infections affecting the bloodstreams, hearts and livers could also occur [35,39,40]. ...
... In more rare occasions especially in severely immunocompromised individuals, disseminated invasive infections affecting the bloodstreams, hearts and livers could also occur [35,39,40]. The aetiological agent for most of the cases were Pleurostoma richardsiae [7-25, 28-33, 35-40], while Pleurostoma ochraceum [26], Pleurostoma ootheca [27] and Pleurostoma repens [34] were also reported to cause infections in a few cases. ...
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The incidence of invasive fungal infections (IFIs) in solid organ transplant (SOT) recipients has increased during the past 20 years and is associated with significant morbidity and mortality. In this post hoc analysis of a large, open-label, multicenter study, we evaluated efficacy and safety of posaconazole, a new extended-spectrum triazole, as salvage therapy for IFIs in SOT recipients. Twenty-three SOT recipients with proven or probable IFI and evidence of disease refractory to, or intolerant of, standard antifungal therapies received posaconazole oral suspension (40 mg/mL) 800 mg daily in divided doses. An independent, blinded data-review committee assessed patient diagnosis and outcome. Complete or partial response was documented in 13 of 23 (57%) SOT recipients with proven or probable IFIs, including 1 of 2 (50%) refractory patients, 5 of 8 (63%) intolerant to prior therapy, and 7 of 13 (54%) who were both. Successes by type of IFI included 7 of 12 with invasive aspergillosis, 2 of 2 with invasive fusariosis, 1 of 1 with cryptococcosis, and 1 of 2 with zygomycosis. Treatment-related adverse events (TRAEs) were reported in 12 of 23 patients. Severe TRAEs occurred in 4 of 23 patients including increased levels of cyclosporine or tacrolimus requiring immunosuppressive dose adjustments in three patients and in one, termination of posaconazole. Severe TRAEs associated with renal and liver toxicities were uncommon. Posaconazole was well tolerated and effective against IFIs including invasive aspergillosis, zygomycosis, fusariosis, and cryptococcosis in SOT recipients intolerant of or failing other antifungal therapies. Calcineurin inhibitor levels should be closely monitored in patients treated concomitantly with posaconazole to avoid toxicity from drug interaction.
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The history and diagnostic characteristics of the Calosphaeriales, family Calosphaeriaceae (Class Ascomycetes) are reviewed. A combination of features of ascomata and asci permits recognition of eight genera: Calosphaeria, Scoptria, Enchnoa, Jattaea, Romellia, Graphostroma, Togninia, and Pleurostoma. Some of the species are described and illustrated. One new species, Romellia tympanoides, is described; the new combinations Scoptria discreta, Jattaea curvicolla, J. ceanothina, Romellia cornina, and Pleurostoma ootheca are proposed.
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Among dematiaceous fungi responsible for phaeohyphomycosis, Cladophialophora bantiana is an opportunistic pathogen that causes central nervous system infections, chiefly in immunocompromised patients. Only a few reports on skin involvements have been reported in the recent dermatological literature. Herein we report the case of an immunocompetent patient with cutaneous phaeohyphomycosis. A 48-year-old male presented a nodular, painless and non-suppurative lesion with a diameter of 1cm on the right buttock that had developed since his return from a trip to Vietnam. A diagnosis of phaeohyphomycosis due to C. bantiana was made based on the histopathology and mycology examinations, which allowed the identification of C. bantiana, a dematiaceous (black) fungus from hyphomycete species. C. bantiana is a neurotropic fungus that causes central nervous system infections in particular. Extracerebral involvement is rare and only a few cases of cutaneous phaeohyphomycosis have been reported. Furthermore, since immunocompromised hosts are more vulnerable, this mycosis is more commonly seen in immunocompromised patients. However in this particular case, an intramuscular injection of corticosteroids could have caused local immunosuppression. The prognosis depends on both localization and site. There are no guidelines for optimal treatment.
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Dematiaceous molds are increasingly recognized as important human pathogens. We report 2 cases of cutaneous phaeohyphomycosis in renal allograft recipients, caused by Alternaria alternata and Curvularia spp., respectively, which demonstrate the diversity in clinical presentation, the different therapeutic strategies, and the clinical importance of azole antifungal-induced drug-drug interactions with immunosuppressive therapy.
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Subcutaneous phaeohyphomycosis in solid organ recipients may have an adverse outcome. We sought to describe the disease course, treatment, and outcome of allograft function in kidney transplant recipients with phaeohyphomycosis. Seventeen patients were followed for a mean period of 25.4 months to analyze the clinical response to treatment. There was no treatment failure or relapsing disease among 12 patients who completed treatment. Two patients were still in treatment with disease remission. One patient discontinued the study during treatment with partial remission, one died after finishing treatment with disease remission, and one was dropped from the study because contact was lost. Immunosuppressive regimens were not changed. Two of 17 patients had a significant reduction in allograft function. The follow-up time was short and the number of patients was small. The outcome of phaeohyphomycosis in kidney transplant recipients was favorable with minimal impact on renal allograft function.
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Nineteen patients with phaeohyphomycosis were treated with itraconazole. Of these, 17 were assessable for clinical outcome. Of these, two had received no prior therapy, five had failed amphotericin B therapy, four had failed ketoconazole or miconazole therapy, and five had failed both amphotericin B and azole therapy. One patient had received only prior surgical intervention. Fungi of seven different genera caused disease of the skin in nine patients, soft tissue in nine, sinuses in eight, bone in five, joints in two, and lungs in two. Itraconazole was given in dosages ranging from 50 to 600 mg/day for 1 to 48 months. Clinical improvement or remission occurred in nine patients. Two patients have had stabilization of disease. Six patients failed treatment, one had a relapse after initially successful treatment. Itraconazole appears to be highly effective in some patients with phaeohyphomycosis, including patients refractory to other antifungal agents.
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A case of phaeohyphomycosis caused by Phialophora richardsiae is presented. The patient was a 30-year-old man with end stage malignant lymphoma. A subcutaneous abscess that developed on the dorsum of the right foot was removed surgically but immediately recurred. The causative mold was isolated from the pus in the lesion and identified as P. richardsiae. This is the fifth known case of phaeohyphomycosis caused by P. richardsiae and the first recorded in Japan.
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