Case Report: Invasive Sinusitis due to Sporothrix Brasiliensis in a Renal Transplant Recipient.

Sporotrichosis is usually a subcutaneous infection caused by thermodimorphic fungi of the genus Sporothrix. The disease occurs worldwide, but endemic areas are located in tropical and subtropical regions. The epidemiology of sporotrichosis in Brazil is peculiar because of the cat's entry in the chain of transmission of this mycosis, associated with Sporothrix brasiliensis, the most virulent species in the genus. Sinusitis caused by Sporothrix species is unusual and may be underdiagnosed or confused with other fungal etiologies, like mucormycosis. We report a case of sinusitis due to a Sporothrix species in a 6-year renal transplant recipient. Direct examination of smears of exudate of the sinus specimen (aspirate, biopsy) revealed budding yeasts and cigar-shaped cells. Sporothrix was subsequently recovered from the patient's exudate culture and identified as S. brasiliensis using species-specific polymerase chain reaction, and she was successfully treated with antifungal therapy. Her parents also developed the disease a week later, both only cutaneous involvement. Sporotrichosis sinusitis is a rare disease, even in immunocompromised patients. Diagnosis is crucial, and benefits from good epidemiological history.

The spread of cat-transmitted sporotrichosis due to Sporothrix brasiliensis in Brazil towards the Northeast region.

BACKGROUND: Sporotrichosis is a worldwide subcutaneous mycosis caused by Sporothrix spp. In the past, this infection was associated with armadillo hunting, horticulturists, miners, and gardeners, being considered an implantation mycosis acquired by plant debris injury. Nevertheless, since the late nineties, it has been considered a zoonotic disease in Brazil. Here we report a case series of 121 patients with cat-transmitted sporotrichosis seen in Northeast Brazil. METHODOLOGY/PRINCIPAL FINDINGS: Patient's demographic, clinical data, and length of treatment were recorded. In addition, a mycological examination and further PCR confirmation of species identification were performed. One hundred and twenty two patients were diagnosed with subcutaneous sporotrichosis from October 2016 to December 2019, while PCR revealed that 71 of them were due to S. brasiliensis. The majority of the individuals were female (n = 86; 70.5%). Patient's age ranged from 5 to 87 years old. The clinical forms found were lymphocutaneous (58.2%) and fixed cutaneous (39.4%). Interestingly, 115 patients reported previous contact with cats diagnosed with sporotrichosis. Patients were successfully treated with itraconazole and potassium iodide. CONCLUSIONS/SIGNIFICANCE: Our study adds important contributions for the investigation of the spread of cat-transmitted subcutaneous sporotrichosis in Brazil, specifically towards the Northeast region of a continental-size country. It will also help clinicians to be aware of the existence and importance to accurately diagnose sporotrichosis and treat patients with this infectious disease in the lowest income region of Brazil.

Clinical and epidemiological aspects of feline sporotrichosis caused by Sporothrix brasiliensis and in vitro antifungal susceptibility.

Sporotrichosis is a subcutaneous mycosis resulting from the traumatic implantation of pathogenic Sporothrix species. In Brazil, zoonotic transmission plays an important role in the epidemiology of the disease, involving especially cats. The objective of this study was to isolate Sporothrix spp. from cats with signs of sporotrichosis, determining the causative species, clinical and epidemiological aspects, and the in vitro susceptibility profile of the isolates against antifungal drugs. From September 2017 to February 2019, 245 samples of lesions were collected from symptomatic cats in São José do Rio Preto, Brazil. Identification of the isolates was performed by morphophysiological parameters and species-specific polymerase chain reaction. The susceptibility profile of the isolates was determined for five drugs (amphotericin B, itraconazole, ketoconazole, potassium iodide and terbinafine), using the broth microdilution method. Clinical and epidemiological aspects were analyzed based on data contained on investigation forms filled by the veterinarians at moment of collection. Sporothrix spp. were isolated in 189 (77.2%) of the samples. Phenotypic and molecular analyses revealed S. brasiliensis as the only causative agent. In vitro susceptibility testing showed lower MIC values for terbinafine (MIC = 0.03-2 µg/ml), ketoconazole (MIC = 0.03-2 µg/ml), and itraconazole (MIC = 0.03-4 µg/ml). Most of the animals were male (73.5%), adults (96.3%), stray (53.5%), and uncastrated (69.8%). Our results show the expansion of the S. brasiliensis epidemic to an area nearly 840 km apart from the epicenter of the long-lasting outbreak of cat-transmitted sporotrichosis in Rio de Janeiro.

Can cats cause colossal contagious cutaneous carbuncles?

Sporothrix spp. cause the most common deep fungal skin infections in Brazil and this is related to infected cats. Transmission is traditionally from organic material/plants but can also be zoonotic. Culture of a skin biopsy is the golden standard for determination. Treatment with oral itraconazole approaches up to 95% efficacy in patients with cutaneous sporotrichosis.

Esporotricosis, una realidad aún presente en Chile: a propósito de un caso / Sporotrichosis, a reality still present in Chile: a case report

OBJETIVO: Reportar un caso de esporotricosis linfocutánea contraída en Chile y realizar una revisión de la literatura con énfasis en los hallazgos dermatoscópicos de las lesiones y en los casos reportados en el país. CASO CLÍNICO: Paciente adulto mayor con lesiones nodulares ulceradas siguiendo trayecto linfático en extremidad superior derecha. RESULTADOS: Estudio micológico confirmó la presencia de Sporothrix schenckii. Se inició tratamiento con antifúngico oral con buena respuesta terapéutica. CONCLUSIONES: Existe un escaso número de reporte de esporotricosis linfocutánea contraída en Chile con confirmación micológica. Este es el único caso chileno con descripción dermatoscópica de las lesiones.

OBJETIVE: To report a case of lymphocutaneous sporotrichosis adquired in Chile and provide a review of the literature with emphasis on dermoscopic findings and previous cases reported in the country. CLINICAL CASE: An elderly patient presented with ulcerated nodular lesions following lymphatic vessels in his right upper limb.RESULTS: Mycological work-up confirmed the presence of Sporothrix schenkii. Oral antifungal treatment was initiated with good therapeutic response. CONCLUSIONS: Few case reports of lymphocutaneous sporotricosis adquired in Chile can be found in literature. Until now this is the only chilean case with dermoscopic description of the lesions.

Canine sporotrichosis: polyphasic taxonomy and antifungal susceptibility profiles of Sporothrix species in an endemic area in Brazil.

Sporotrichosis, a mycosis caused by pathogenic species of the genus Sporothrix, affects diverse species of mammals. Until 2007, Sporothrix schenckii was considered the unique etiologic agent of sporotrichosis. Canine sporotrichosis is a poorly reported disease, and the majority of cases are from Rio de Janeiro, Brazil. There are scarce studies on the characterization of canine isolates of Sporothrix schenckii complex, as well as few antifungal susceptibility data available. The aim of this study was to characterize the clinical isolates of Sporothrix from dogs from Brazil at species level and evaluate their antifungal susceptibility profile. Polyphasic taxonomy was used to characterization at species level (morphological, phenotypical characteristics, and molecular identification). Antifungal susceptibility profiles (amphotericin B, itraconazole, ketoconazole, posaconazole, and terbinafine) were determined using the Clinical and Laboratory Standards Institute broth microdilution method (M38-A2). According to phenotypic identification and molecular analysis, 46 isolates included in this study were identified as S. brasiliensis and one as S. schenckii. Amphotericin B presented the highest minimum inhibitory concentration values, and the other drugs showed effective in vitro antifungal activity. This is the first report of S. schenckii in dogs from Brazil, since S. brasiliensis is the only species that has been described in canine isolates from Rio de Janeiro to date. Nevertheless, no differences were observed in the antifungal susceptibility profiles between the S. brasiliensis and S. schenckii isolates, and it is important to continuously study new canine clinical isolates from Rio de Janeiro, Brazil.

Geographical distribution and ecological niche modeling of the etiological agents of human sporotrichosis in Venezuela.

The geographical distribution and ecological niche of the two circulating species of the Sporothrix genus in Venezuela was established. For this, 68 isolates of Sporothrix spp. from patients of different regions of the country were analyzed. A molecular taxonomy analysis was conducted using a fragment of the calmodulin gene (CAL), and ITS regions, confirming the presence of S. schenckii (62%) and S. globosa (38%). Computational models of ecological niche for each species were obtained by the maximum entropy method using the MaxEnt software, which predicted the best environmental conditions for the presence of the two species. These models predict that the main variables influencing the presence of S. schenckii were altitude and annual mean temperature, while for S. globosa, the more influent variable was the land use, with 82% of S. globosa located at urban areas vs 56% for S. schenckii. The results here presented could contribute to understand the specific environmental factors that might modulate the occurrence of Sporothrix spp. as well as its transmission. To our knowledge, our analyses show for the first time Sporothrix spp.-specific ecological niche data, a valuable tool to promote evidence-based public health policymaking within endemic areas of sporotrichosis.

Phenotypic and molecular characterisation of Sporothrix globosa of diverse origin from India.

Sporotrichosis is one of the neglected tropical diseases causing subcutaneous chronic granulomatous lesion by thermally dimorphic fungi belonging to Sporothrix species. Sporothrix brasiliensis, Sporothrix mexicana and Sporothrix globosa are the common pathogenic species. In Asian countries, S. globosa constitutes nearly 99.3% of all Sporothrix species. We studied 63 cases of sporotrichosis of geographically diverse origin from India and Sporothrix isolates were characterised for its growth in different media, temperatures, ability to assimilate sugars and antifungal susceptibility profile. Molecular characterization was performed by sequencing of the calmodulin (CAL), beta tubulin (BT) and translational elongation factor 1-alpha (TEF-1α) and typing by fluorescent amplified fragment length polymorphism (FAFLP). In patients who presented with fixed (49.2%), lymphocutaneous lesions (23.8%), in 26.9% the details were not known, none had systemic dissemination. All the isolates tested were Sporothrix globosa and that could grow up to 35 °C and unable to grow at and beyond 37 °C. The assimilation of sucrose, ribitol and raffinose helps in identifying S. globosa. Sequences of CAL or BT or TEF-1α can differentiate S. globosa from other species in the complex. FAFLP results exhibited low genetic diversity. No correlation was noted between genotypes and clinical presentation, or geographic distribution. Itraconazole, terbinafine and posaconazole showed good in vitro antifungal activity against S. globosa whereas fluconazole and micafungin had no activity. S. globosa of Indian origin is relatively less pathogenic than other pathogenic Sporothrix species as it does not cause systemic dissemination and in the diagnostic laboratory, incubation of the cultures below 37 °C is essential for effective isolation.

Sporotrichosis in Mexico.

Sporotrichosis is an endemic mycosis caused by the species of the Sporothrix genus, and it is considered one of the most frequent subcutaneous mycoses in Mexico. This mycosis has become a relevant fungal infection in the last two decades. Today, much is known of its epidemiology and distribution, and its taxonomy has undergone revisions. New clinical species have been identified and classified through molecular tools, and they now include Sporothrix schenckii sensu stricto, Sporothrix brasiliensis, Sporothrix globosa, and Sporothrix luriei. In this article, we present a systematic review of sporotrichosis in Mexico that analyzes its epidemiology, geographic distribution, and diagnosis. The results show that the most common clinical presentation of sporotrichosis in Mexico is the lymphocutaneous form, with a higher incidence in the 0-15 age range, mainly in males, and for which trauma with plants is the most frequent source of infection. In Mexico, the laboratory diagnosis of sporotrichosis is mainly carried out using conventional methods, but in recent years, several researchers have used molecular methods to identify the Sporothrix species. The treatment of choice depends mainly on the clinical form of the disease, the host's immunological status, and the species of Sporothrix involved. Despite the significance of this mycosis in Mexico, public information about sporotrichosis is scarce, and it is not considered reportable according to Mexico's epidemiological national system, the "Sistema Nacional de Vigilancia Epidemiológica." Due to the lack of data in Mexico regarding the epidemiology of this disease, we present a systematic review of sporotrichosis in Mexico, between 1914 and 2019, that analyzes its epidemiology, geographic distribution, and diagnosis.

A novel Sporothrix brasiliensis genomic variant in Midwestern Brazil: evidence for an older and wider sporotrichosis epidemic.

Sporotrichosis is a subcutaneous infection caused by fungi from the genus Sporothrix. It is transmitted by inoculation of infective particles found in plant-contaminated material or diseased animals, characterizing the classic sapronotic and emerging zoonotic transmission, respectively. Since 1998, southeastern Brazil has experienced a zoonotic sporotrichosis epidemic caused by S. brasiliensis, centred in the state of Rio de Janeiro. Our observation of feline sporotrichosis cases in Brasília (Midwestern Brazil), around 900 km away from Rio de Janeiro, led us to question whether the epidemic caused by S. brasiliensis has spread from the epicentre in Rio de Janeiro, emerged independently in the two locations, or if the disease has been present and unrecognized in Midwestern Brazil. A retrospective analysis of 91 human and 4 animal cases from Brasília, ranging from 1993 to 2018, suggests the occurrence of both sapronotic and zoonotic transmission. Molecular typing of the calmodulin locus identified S. schenckii as the agent in two animals and all seven human patients from which we were able to recover clinical isolates. In two other animals, the disease was caused by S. brasiliensis. Whole-genome sequence typing of seven Sporothrix spp. strains from Brasília and Rio de Janeiro suggests that S. brasiliensis isolates from Brasília are genetically distinct from those obtained at the epicentre of the outbreak in Rio de Janeiro, both in phylogenomic and population genomic analyses. The two S. brasiliensis populations seem to have separated between 2.2 and 3.1 million years ago, indicating independent outbreaks or that the zoonotic S. brasiliensis outbreak might have started earlier and be more widespread in South America than previously recognized.

Tregs in the immune response of BALB/c mice experimentally infected with species of the Sporothrix genus.

Background: Sporotrichosis occurs through contact with contaminated soil and plant. However, the incidence of sporotrichosis as a zoonotic epidemic has increased, particularly in Rio de Janeiro. Aim: In this work, we decided to evaluate some T-cell phenotypes involved in the immune response. Materials & methods: We used flow cytometry to quantify TCD4+ and TCD8+ and Treg from immunocompetent and immunosuppressed mice infected with Sporothrix species with different levels of virulence and pathogenicity. Results: It was demonstrated the predominance of TCD4+ over the TCD8+ cells in both groups, inoculated with all the species, and percentages of Treg observed in infected immunocompetent mice. Conclusion: This regulatory phenotype can be associated with a protective immunity in the initial periods of infection.

Disseminated sporotrichosis with immune reconstitution inflammatory syndrome in an HIV patient: case report and review of the literature / Esporotricosis diseminada y síndrome inflamatorio de reconstitución inmune en un paciente con VIH: reporte de un caso y revisión de la literatura

BACKGROUND: Sporotrichosis has been occurring as outbreaks in Brazil, reaching epidemic levels in some regions. Zoonotic transmission is the main route to acquire Sporothrix. CASE REPORT: We describe a case of disseminated sporotrichosis caused by Sporothrix brasiliensis in an HIV/AIDS patient, with the presentation of immune reconstitution inflammatory syndrome (IRIS). CONCLUSIONS: This case reinforces that sporotrichosis should always be suspected in patients with IRIS from endemic regions, even in patients without the typical cutaneous lesions of this mycosis

ANTECEDENTES: La esporotricosis suele aparecer en Brasil en forma de brotes y alcanza tasas epidémicas en algunas regiones. La ruta principal de transmisión es la zoonótica. CASO CLÍNICO: Describimos un caso de esporotricosis diseminada causado por Sporothrix brasiliensis en una paciente con VIH/sida que presentó un síndrome inflamatorio de reconstitución inmune (SIRI). CONCLUSIONES: Este caso demuestra que en regiones endémicas de esporotricosis esta micosis siempre debe ser sospechada en casos de SIRI, incluso en pacientes sin las lesiones cutáneas típicas de esta enfermedad

Disseminated cutaneous sporotrichosis: unusual presentation in an alcoholic patient.

Sporotrichosis is an implantation mycosis highly prevalent in Brazil, associated with soil activities and contact with infected animals. It has numerous clinical manifestations and its disseminated cutaneous form is uncommon, found in approximately 4% of cases and usually related to immunosuppressive conditions. We report an alcoholic 56-year-old male patient with no other comorbidities, presenting with multiple cutaneous nodules and ulcers. The patient was diagnosed with disseminated cutaneous sporotrichosis based on the isolation and identification of Sporothrix spp. The patient was initially treated with amphotericin B with satisfactory results and then with itraconazole.

Direct PCR of lesions suggestive of sporotrichosis in felines / [PCR direta de lesões sugestivas de esporotricose em gatos]

A esporotricose é uma doença emergente e a incidência de esporotricose zoonótica tem aumentado, principalmente no Brasil; a maioria dos casos está relacionada à transmissão de gatos infectados. O diagnóstico definitivo da esporotricose felina é feito por cultura fúngica; no entanto, aguardar o longo período de cultura pode atrasar o início do tratamento. O objetivo deste estudo foi detectar e determinar as espécies de Sporothrix por PCR realizado diretamente das lesões de gatos, provenientes de área endêmica, nos quais a esporotricose fazia parte do diagnóstico diferencial. Um total de 87,1% dos casos foi positivo por PCR ou cultura fúngica para Sporothrix; 81,4% foram confirmados como S. brasiliensis por PCR, 71,4% por isolamento e 65,7% pelos dois métodos. Em conclusão, a análise direta por PCR de lesões sugestivas de esporotricose em gatos é um bom método para confirmar a infecção e determinar as espécies de Sporothrix, garantindo um diagnóstico rápido; esse método tem uma boa concordância com o isolamento fúngico.(AU)

Benzylidene-carbonyl compounds are active against itraconazole-susceptible and itraconazole-resistant Sporothrix brasiliensis.

We evaluated the antifungal activity of benzylidene-carbonyl compounds (LINS03) based on the structure of gibbilimbol from Piper malacophyllum Linn. Five analogues (1-5) were synthetized following a classic aldol condensation between an aromatic aldehyde and a ketone, under basic conditions. These were tested against itraconazole-susceptible (n = 3) and itraconazole-resistant (n = 5) isolates of Sporothrix brasiliensis by M38-A2 guidelines of CLSI. All of them were fungistatic (MIC ranged of 0.11-0.22 mg/mL (1); 0.08-0.17 mg/mL (2); 0.05-0.1 mg/mL (3); 0.04-0.33 mg/mL (4); and 0.04-0.3 mg/mL (5)), highlighting compounds 2 and 3. As fungicidal, compounds 1 and 2 were highlighted (MFC ranged of 0.22-0.89 mg/mL and 0.08-1.35 mg/mL, respectively), compared with the remaining (0.77-> 3.08 mg/mL (3); 0.08-> 2.6 mg/mL (4); and 0.59-> 2.37 mg/mL (5)). The inhibitory activity was related to the benzylidene-carbonyl, whereas the phenol group and the low chain homolog seems to contribute to some extent to the fungicidal effect. Compound 2 highlighted due to the considerable fungistatic and fungicidal activities, including itraconazole-resistant Sporothrix brasiliensis. These findings support the potential usefulness of benzylidene-carbonyl compounds as promising prototypes for the development of antifungal against sporotrichosis by Sporothrix brasiliensis, including against itraconazole-resistant isolates.

5-Aminolevulinic acid-mediated photodynamic therapy has effective antifungal activity against Sporothrix globosa in vitro.

BACKGROUND: An alternative therapy for sporotrichosis is necessary to reduce the treatment time and raise clinical efficacy. The 5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) is a promising platform with which to treat mycoses. However, despite the worldwide prevalence of Sporothrix globosa, a causative agent of sporotrichosis, the effect of ALA-PDT on this pathogen has not been validated. OBJECTIVES: The aim of this study was to evaluate the effect of ALA-PDT on S globosa and the protection of melanin through an in vitro study. The mechanisms involved were also investigated. METHODS: To estimate the survival rate of S globosa treated with ALA-PDT and the protection offered by melanin, the conidia and yeast cells of wild-type S globosa (Mel+), other clinical strains, tricyclazole-treated Mel+ and an albino mutant strain (Mel-) were incubated with and without ALA or irradiation. Reactive oxygen species generation by Mel+ conidia induced by ALA-PDT was assayed. SEM and TEM were conducted to obverse ultrastructural changes in the conidia. A comet assay was performed to evaluate DNA damage. RESULTS: The survival rate of S globosa conidia and yeast cells significantly decreased following incubation with 1.19M ALA and 162 J/cm2 irradiation in vitro. Melanin was not only capable of protecting the conidia against ALA-PDT, but also against ALA or irradiation alone. After induction by ALA-PDT, alterations in reactive oxygen species generation, DNA damage and ultrastructural changes were observed. CONCLUSIONS: ALA-PDT inhibits the survival of S globosa conidia in vitro and therefore has potential for the treatment of sporotrichosis.

Genome-wide mapping using new AFLP markers to explore intraspecific variation among pathogenic Sporothrix species.

Sporotrichosis is a chronic subcutaneous mycosis caused by Sporothrix species, of which the main aetiological agents are S. brasiliensis, S. schenckii, and S. globosa. Infection occurs after a traumatic inoculation of Sporothrix propagules in mammals' skin and can follow either a classic route through traumatic inoculation by plant debris (e.g., S. schenckii and S. globosa) or an alternative route through zoonotic transmission from animals (e.g., S. brasiliensis). Epizootics followed by a zoonotic route occur in Brazil, with Rio de Janeiro as the epicenter of a recent cat-transmitted epidemic. DNA-based markers are needed to explore the epidemiology of these Sporothrix expansions using molecular methods. This paper reports the use of amplified-fragment-length polymorphisms (AFLP) to assess the degree of intraspecific variability among Sporothrix species. We used whole-genome sequences from Sporothrix species to generate 2,304 virtual AFLP fingerprints. In silico screening highlighted 6 primer pair combinations to be tested in vitro. The protocol was used to genotype 27 medically relevant Sporothrix. Based on the overall scored AFLP markers (97-137 fragments), the values of polymorphism information content (PIC = 0.2552-0.3113), marker index (MI = 0.002-0.0039), effective multiplex ratio (E = 17.8519-35.2222), resolving power (Rp = 33.6296-63.1852), discriminating power (D = 0.9291-0.9662), expected heterozygosity (H = 0.3003-0.3857), and mean heterozygosity (Havp = 0.0001) demonstrated the utility of these primer combinations for discriminating Sporothrix. AFLP markers revealed cryptic diversity in species previously thought to be the most prevalent clonal type, such as S. brasiliensis, responsible for cat-transmitted sporotrichosis, and S. globosa responsible for large sapronosis outbreaks in Asia. Three combinations (#3 EcoRI-FAM-GA/MseI-TT, #5 EcoRI-FAM-GA/MseI-AG, and #6 EcoRI-FAM-TA/MseI-AA) provide the best diversity indices and lowest error rates. These methods make it easier to track routes of disease transmission during epizooties and zoonosis, and our DNA fingerprint assay can be further transferred between laboratories to give insights into the ecology and evolution of pathogenic Sporothrix species and to inform management and mitigation strategies to tackle the advance of sporotrichosis.

Infection caused by Sporothrix schenckii: an autochthonous case in Bari, Southern Italy.

An autochthonous case of lymphocutaneous sporotrichosis caused by Sporothrix schenckii is reported. The patient developed skin lesions localized along the lymphatics that appeared after he suffered an injury while collecting wicker canes in marshy water. The fungus was identified as Sporothrix schenckii by MALDI-TOF and sequencing. Phylogenetic analysis was also performed. Low MIC values were detected for all tested echinocandins and azoles except for fluconazole. The patient was treated with itraconazole without significant improvement. A regression of lesions was observed after 3 months of therapy with voriconazole. Few cases of sporotrichosis have been reported in Europe. However, several cases of sporotrichosis have been described in Italy. The incidence of sporotrichosis in Italy may be underestimated and microbiologists, and clinicians must be aware of this fungal infection.