A comparative study of extracellular enzymes from chromoblastomycosis agents reveals the potential association of phospholipase with the severity of the lesions.

Chromoblastomycosis is a chronic, progressive subcutaneous mycosis that is endemic in tropical and subtropical countries. Cladophialophora carrionii and Fonsecaea pedrosoi are prevalent etiological agents. The potential role of the proteolytic activity of extracellular enzymes in these fungi and its relationship with the pathogenesis of the disease has not been proven. Some phenotypic traits have been associated with the virulence of other fungi; i.e., their different rate of protease, phospholipase, and esterase excretion, melanin, and thermotolerance. The aim of this study was the identification of extracellular enzymes that could be considered virulence markers of chromoblastomycosis agents. Therefore, we tested 29 C. carrionii and 11 F. pedrosoi clinical isolates to determine their hydrolytic and physiologic characteristics. All the tested isolates grew at a range of 30°-37 °C; except 2 strains of F. pedrosoi that grew slowly at 40 °C. We noticed that the hydrolytic capabilities of the tested isolates were positive for urea hydrolysis in almost all, while both strains were negative for DNase, hemolysin, and gelatin. C. carrionii and F. pedrosoi had phospholipase and esterase activity. These findings were similar for most isolates. All strains showed an association between phospholipase activity and moderate to severe lesions. However, only in F. pedrosoi isolates, the association remains significant. We conclude that the different enzymatic production reported here may be linked to the clinical manifestations of these pathologies. Notwithstanding, the influence of other virulence factors is not excluded.

In vitro susceptibility of chromoblastomycosis agents to five antifungal drugs and to the combination of terbinafine and amphotericin B.

Chromoblastomycosis is a chronic mycosis that affects the skin and subcutaneous tissues caused by several genera of dematiaceous fungi. There is not a treatment of choice. Thus, tools that help guide clinical practice are fundamental. In this sense, antifungal activity tests in vitro could be useful. However, trials with chromoblastomycosis agents are scarce. The aim of this study was to evaluate both the in vitro susceptibility of 60 chromoblastomycosis agents to five antifungals and the combination of amphotericin B (AMB) and terbinafine (TRB). TRB, itraconazole (ITZ) and ketoconazole (KTZ) were, in this order, the drugs which showed better activity against the chromoblastomycosis agents. The less active drugs were voriconazole (VRZ) and AMB. The more differentiated group was Exophiala spinifera. Cladophialophora carrionii and Fonsecaea spp. are significantly more susceptible to KTZ than Phialophora verrucosa, whereas C. carrionii is significantly more sensitive to VRZ than P. verrucosa and E. spinifera. Assays in this direction allow the knowledge of the susceptibility of the causative agents which may help the management of patients with this disease. This study includes the largest number of these agents and of genera found in the literature.

Galleria mellonella in vitro model for chromoblastomycosis shows large differences in virulence between isolates.

BACKGROUND: Chromoblastomycosis is the World Health Organization (WHO)-recognized fungal implantation disease that eventually leads to severe mutilation. Cladophialophora carrionii (C. carrionii) is one of the agents. However, the pathogenesis of C. carrionii is not fully investigated yet. METHODS: We investigated the pathogenic potential of the fungus in a Galleria mellonella (G. mellonella) larvae infection model. Six strains of C. carrionii, and three of its environmental relative C. yegresii were tested. The G. mellonella model was also applied to determine antifungal efficacy of amphotericin B, itraconazole, voriconazole, posaconazole, and terbinafine. RESULTS: All strains were able to infect the larvae, but virulence potentials were strain-specific and showed no correlation with clinical background of the respective isolate. Survival of larvae also varied with infection dose, and with growth speed and melanization of the fungus. Posaconazole and voriconazole exhibited best activity against Cladophialophora, followed by itraconazole and terbinafine, while limited efficacy was seen for amphotericin B. CONCLUSION: Infection behavior deviates significantly between strains. In vitro antifungal susceptibility of tested strains only partly explained the limited treatment efficacy in vivo.

Chromoblastomycosis in Santa Catarina state, Brazil / Cromoblastomicosis en el estado de Santa Catarina, Brasil

A case of chromoblastomycosis caused by Cladophialophora carrionii is reported. The diabetic and hypertensive patient presented serpiginous and verrucous lesions, with centrifugal evolution. The patient, with a history of disease for 59 years, had not been diagnosed or treated before. Dematiaceous septate hyphal and elliptical conidia were seen on microscopic observations. The isolated fungus was identified on the basis of micro-macromorphologic characteristics.

Se reportó un caso de cromoblastomicosis causado por Cladophialophora carrionii. El paciente, diabético e hipertenso, presentaba lesiones de apariencia verrugosa y serpiginosa, con evolución centrífuga. Tenía un historial de enfermedad hace 59 años sin haber sido diagnosticado hasta entonces. En el examen microscópico de observaron elementos hifales septados, pigmentados y con conidios elípticos. La identificación del hongo se basó en las características macromorfológicas y micromorfológicas.

Cromomicosis: una endemia rural en la región noroccidental en Venezuela / Chromomycosis: rural endemic in the north-western region of Venezuela

INTRODUCCIÓN: la cromomicosis, descrita por Max Rudolph en 1914, es una micosis subcutánea de desarrollo crónico, que se adquiere generalmente a partir de un traumatismo cutáneo y es causada por hongos dematíaceos. En Venezuela se ha reportado en todo el país aunque hay franco predominio en los estados de Lara, Zulia y Falcón. OBJETIVO: establecer algunos factores que pudieran explicar la endemia en la zona rural del estado de Falcón, Venezuela. MÉTODOS: se procedió a la detección in vitro de Cladophialophora carrionii a partir de la vegetación típica de esta región, semiárida, descrita como "bosque xerófilo de espinar". Se realizó además, búsqueda activa de casos clínicos entre los habitantes de la zona, mediante examen directo y cultivo de las escamas obtenidas a partir de lesiones y se trató de establecer, la existencia un factor de susceptibilidad hereditario mediante el cálculo del factor de heredabilidad de Falconer y la metodología de problemas genéticos heredables de Arias. RESULTADOS: se pudo aislar C. carrionii a partir de especies xerófilas abundantes en la zona: Prosopis juliflora; y varias cactáceas. La casuística acumulada, en nuestro estado representa 54,4 % (490/900) de todos los casos reportados en Venezuela, desde 1983 hasta 2005. La mayoría de los pacientes fueron infectados por C. carrionii y refirieron traumas frecuentes con espinas de cactáceas. Mediante estudios genealógicos se demostró la concentración de los casos en grupos familiares (hasta 11 % superior a la comunidad) así como un factor de heredabilidad de 65 % y alta frecuencia de matrimonios consanguíneos (25 %) asociados con 7 % de personas con cromomicosis. CONCLUSIONES: se consideró que se trata de una endemia rural, laboral y familiar, posiblemente relacionada con un factor de susceptibilidad heredable.

INTRODUCTION: chromomycosis, described by Max Rudolph in 1914, is a chronic subcutaneous mycosis that are generally caught from a cutaneous trauma and caused by dematiaceous fungi. This disease has been reported throughout Venezuela, but it is predominant in the states of Lara, Zulia and Falcón. OBJECTIVE: to ascertain some factors that might explain the endemic at Falcon State rural zone. METHODS: in vitro detection of Cladophialophora carrionii from the typical vegetation of this semiarid area known as "xerophilous woods of prickles" and also, active search of clinical cases among inhabitants of this area through direct testing and culture of flakes from skin lesions. It was intended to determine a hereditary susceptibility factor through the estimation of Falconer´s heritability factor and Arias´ methodology of heritable genetic disorders. RESULTS: it was possible to isolate C. carrionii from abundant xerophilous species in the area such as Prosopis juliflora and several Cactaceae. The accumulated casuistry in our state accounts for 54,4% (490/900) of all cases reported in Venezuela from 1983 to 2995. Most of the patients were infected by C. carrionii and they mentioned frequent injures by cactaceae prickles. On the basis of genealogical studies, it was proved that cases were concentrated in family groups (up to 11% higher than in the community) as well as 65% heritability factor and high frequency of blood-related marriages (25%) associated with 7% of people affected by chromomycosis. CONCLUSIONS: it was concluded that this is a rural, work and family endemic that is likely associated with a hereditary susceptibility factor.

Cladophialophora carrionii: a rare agent of chromoblastomycosis in Rio de Janeiro State, Brazil / Cladophialophora carrionii: agente raro de cromoblastomicose no estado do Rio de Janeiro, Brasil

A 73 year-old male farm laborer from a rural area presented a 15 year history of extensive tumoral lesions over his left leg. Histological studies of skin biopsy showed pseudoepitheliomatous hyperplasia and granulomatous chronic inflammatory process with muriform cells, confirming chromoblastomycosis (CBM). Cladophialophora carrionii was isolated in culture. Treatment with itraconazole 400 mg/day for 12 months resulted in complete remission of lesions. As far we aware, this is the first case report of CBM caused by Cladophialophora carrionii in Rio de Janeiro State, Brazil.

Lavrador, com 73 anos, residente em área rural apresentava há 15 anos lesões tumorais disseminadas na perna esquerda. Exame histopatológico de biópsia de pele mostrou hiperplasia pseudo-epiteliomatosa e processo inflamatório crônico granulomatoso com células muriformes, confirmando o diagnóstico de cromoblastomicose (CBM). Cladophialophora carrionii foi isolado na cultura. Tratamento com itraconazol 400 mg/dia durante 12 meses resultou na completa remissão das lesões. Este é o primeiro relato de CBM causado por C. carrionii no estado do Rio de Janeiro, Brasil.

Respuestas inflamatorias y de anticuerpos contra Cladophialophora carrionii en caprinos / Inflamatory and Antibody Responses against Cladophialophora carrionii in Goats

Cladophialophora carrionii es un hongo dimórfico con melanina que causa en humanos una micosis profunda, localizada y crónica llamada cromomicosis, y se adquiere a través de la inoculación con espinas de cactáceas infectadas. El estado Falcón es una zona endémica para esta enfermedad, donde las poblaciones están integradas principalmente por campesinos criadores de caprinos. El objetivo de este trabajo fue evaluar la respuesta inmunitaria humoral e inflamatoria contra C. carrionii en caprinos con el fin de conocer la interacción hospedador-parásito en un animal usualmente considerado como resistente a la infección, a pesar de su exposición permanente a las espinas infectadas. Se inocularon seis cabritos con 1×10(6) células de C. carrionii, vía subcutánea, en la tabla del cuello. Se midió la temperatura corporal de cada animal y se tomaron muestras de sangre completa y suero para análisis hematológicos y ensayos inmunoenzimáticos (ELISA) dos veces a la semana. Se midió el diámetro de la piel 24 y 48 horas después de la inoculación. La respuesta inflamatoria en el sitio de inoculación fue notoria a las 48 horas. No hubo respuesta significativa de anticuerpos en los animales inoculados a partir de los 50 días post-inoculación. Se confirma que la respuesta celular es importante en la resistencia contra C. carrionii, pero la respuesta de anticuerpos parece no ser relevante en esta especie animal.

Cladophialophora carrionii is a dimorphic fungus with melanin which causes a deep, localized and chronic mycosis in humans, named chromomycosis, acquired through inoculation with cactuses infected-spines. Falcon state is an endemic zone to this disease, where human populations make use of goat breeding as their main way of living. The objective of this work was to evaluate both humoral immune and inflammatory responses against C. carrionii in goats to improve knowledge about host-parasite interactions in a species usually considered as infection-resistant, in spite of permanent exposition to infected spines. Six kids were inoculated with 1 ×10(6) cells of C. carrionii, by subcutaneous route on the neck. Blood and sera samples for hematological and immunoenzimatic tests were taken and body temperature was also measured. The diameter of the inoculated skin was also evaluated 24 and 48 hours post-inoculation. Inflammatory response was considerable at 48 hours. There was no valuable response of antibodies. It is established that inflammatory response is important in resistance against C. carrionii, but antibody response seems not to be relevant for this animal specie.