RESUMEN

INTRODUCTION: Sporotrichosis is a chronic infectious mycosis caused by traumatic implantation of Sporothrix spp., which poses a significant challenge to public health because of its zoonotic nature. It affects humans and other animals, particularly cats. This study investigated epidemiological trends and spatial distribution of feline sporotrichosis in Espírito Santo between 2017 and 2022. METHODS: This study collected diagnostic data on sporotrichosis from the Veterinary Pathology Laboratory and Center for Infectious Diseases in Espírito Santo from 2017 to 2022. Statistical analyses were performed using STATA and spatial analyses were performed using ArcGIS software to create maps and identify hotspots. Kernel density estimation and directional mean statistics were applied to visualise the disease concentration and transmission trends. RESULTS: The results showed a significant increase in feline sporotrichosis cases in Espírito Santo from 2017 to 2022, with hotspots identified in urban and densely populated areas. Anchieta, Aracruz and Vila Velha reported the highest numbers of cases. Spatial analysis indicated a spread towards the central and northern coastal regions. CONCLUSION: This study highlights the growing threat of feline sporotrichosis in Espírito Santo, Brazil, particularly in densely populated urban areas. This underscores the urgent need for comprehensive public health strategies that address urbanisation, human-animal interactions and managing stray cat populations. Effective control measures and enhanced surveillance are crucial to mitigate the spread of this zoonotic disease. Importantly, this study provides valuable insights into the epidemiology of sporotrichosis in Brazil and offers a framework for other regions identified as emerging hotspots that face similar challenges.

RESUMEN

Over the past two decades, zoonotic sporotrichosis transmitted by naturally infected cats has become hyperendemic in Rio de Janeiro, Brazil. Sporothrix brasiliensis is the main agent involved. However, there are other forms of transmission of sporotrichosis. The aim of this study was to evaluate and associate the epidemiological, clinical and therapeutic data and the susceptibility of Sporothrix spp. to antifungal drugs in 43 non-zoonotic sporotrichosis cases. Forty-three clinical strains of Sporothrix were identified by partial sequencing of the calmodulin gene. An antifungal susceptibility test of amphotericin B, terbinafine, itraconazole, posaconazole and isavuconazole was performed according to the broth microdilution method. Most patients were male (55.8%). Regarding the source of infection, 21 patients (48.8%) reported trauma involving plants and/or contact with soil. Sporothrix brasiliensis was the predominant species (n = 39), followed by S. globosa (n = 3) and S. schenckii (n = 1). Sporothrix brasiliensis was associated with all the sources of infection, reinforcing previous data showing the presence of this species in environmental sources, as well as with all the clinical forms, including severe cases. One clinical strain of Sporothrix brasiliensis was classified as a non-wild-type strain for amphotericin B and another for itraconazole. S. schenckii was classified as non-WT for all the antifungals tested. In this context, it is important to emphasize that non-zoonotic sporotrichosis still occurs in the state of Rio de Janeiro, with S. brasiliensis as the main etiological agent, primarily associated with infections acquired after traumatic inoculation with plants and/or soil contact, followed by S. globosa and S. schenckii. In addition, non-WT strains were found, indicating the need to monitor the antifungal susceptibility profile of these species. It is crucial to investigate other natural sources of S. brasiliensis to better understand this fungal pathogen and its environment and host cycle.

RESUMEN

A 78-year-old woman presented to hospital with altered mental status. Hyponatremia was diagnosed and treated. She maintained mental confusion despite normal sodium. No headache, fever or focal signs were present. CSF analysis showed chronic meningitis; MRI demonstrated basilar enhancement. She was empirically treated for tuberculosis, with no improvement. PCR for Sporothrix in the CSF was positive. After treatment with Amphotericin followed by oral itraconazole, she completely recovered her cognitive abilities. Follow-up CSF was normal. This report illustrates the need to consider sporotrichosis in the differential diagnosis of chronic meningitis in immunocompetent, and the importance of PCR as a diagnostic tool.

RESUMEN

Sporotrichosis, the cutaneous mycosis most commonly reported in Latin America, is caused by the Sporothrix clinical clade species, including Sporothrix brasiliensis and Sporothrix schenckii sensu stricto. Due to its zoonotic transmission in Brazil, S. brasiliensis represents a significant health threat to humans and domestic animals. Itraconazole, terbinafine, and amphotericin B are the most used antifungals for treating sporotrichosis. However, many strains of S. brasiliensis and S. schenckii have shown resistance to these agents, highlighting the importance of finding new therapeutic options. Here, we demonstrate that milteforan, a commercial veterinary product against dog leishmaniasis, whose active principle is miltefosine, is a possible therapeutic alternative for the treatment of sporotrichosis, as observed by its fungicidal activity in vitro against different strains of S. brasiliensis and S. schenckii. Fluorescent miltefosine localizes to the Sporothrix cell membrane and mitochondria and causes cell death through increased permeabilization. Milteforan decreases S. brasiliensis fungal burden in A549 pulmonary cells and bone marrow-derived macrophages and also has an immunomodulatory effect by decreasing TNF-α, IL-6, and IL-10 production. Our results suggest milteforan as a possible alternative to treat feline sporotrichosis. IMPORTANCE: Sporotrichosis is an endemic disease in Latin America caused by different species of Sporothrix. This fungus can infect domestic animals, mainly cats and eventually dogs, as well as humans. Few drugs are available to treat this disease, such as itraconazole, terbinafine, and amphotericin B, but resistance to these agents has risen in the last few years. Alternative new therapeutic options to treat sporotrichosis are essential. Here, we propose milteforan, a commercial veterinary product against dog leishmaniasis, whose active principle is miltefosine, as a possible therapeutic alternative for treating sporotrichosis. Milteforan decreases S. brasiliensis fungal burden in human and mouse cells and has an immunomodulatory effect by decreasing several cytokine production.

Asunto(s)

Antifúngicos , Enfermedades de los Gatos , Sporothrix , Esporotricosis , Animales , Esporotricosis/tratamiento farmacológico , Esporotricosis/microbiología , Esporotricosis/veterinaria , Gatos , Sporothrix/efectos de los fármacos , Antifúngicos/farmacología , Enfermedades de los Gatos/tratamiento farmacológico , Enfermedades de los Gatos/microbiología , Humanos , Fosforilcolina/análogos & derivados , Fosforilcolina/farmacología , Fosforilcolina/uso terapéutico , Brasil , Pruebas de Sensibilidad Microbiana , Perros , Macrófagos/efectos de los fármacos , Macrófagos/microbiología , Ratones

RESUMEN

Zoonotic sporotrichosis caused by Sporothrix brasiliensis is an emerging mycosis in Latin America. One of the problems to quickly treat infected animals and break the transmission chain is associated with the time-consuming gold-standard diagnosis method (culture). We aimed to evaluate a species-specific polymerase chain reaction (PCR) for the diagnosis of sporotrichosis caused by S. brasiliensis using non-invasive samples. We performed a retrospective cross-sectional study using samples collected with swabs from humans and cats with clinical suspicion of sporotrichosis. Deoxyribonucleic acid (DNA) was extracted using a commercial kit, and a species-specific PCR for S. brasiliensis detection was performed. One hundred ten samples were included. PCR showed a good concordance with culture (86% of agreement) for human and cat samples (Kappa coefficient = 0.722, and 0.727, respectively). In conclusion, our data shows that this adapted PCR using non-invasive samples can be applied to sporotrichosis diagnosis, being a good alternative mainly in regions with a lack of mycologists to identify the fungus in culture, contributing to the control of this emergent zoonosis.

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We aimed to evaluate a molecular method for diagnosing sporotrichosis caused by Sporothrix brasiliensis in humans and cats. We observed that the technique is in good agreement with the classic method and is a good alternative for assisting in the diagnosis and consequent control of this zoonosis.

Asunto(s)

Enfermedades de los Gatos , Reacción en Cadena de la Polimerasa , Sporothrix , Esporotricosis , Esporotricosis/diagnóstico , Esporotricosis/microbiología , Esporotricosis/veterinaria , Gatos , Sporothrix/genética , Sporothrix/aislamiento & purificación , Sporothrix/clasificación , Humanos , Animales , Reacción en Cadena de la Polimerasa/métodos , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/microbiología , Estudios Retrospectivos , Estudios Transversales , Zoonosis/diagnóstico , Zoonosis/microbiología , ADN de Hongos/genética , Técnicas de Diagnóstico Molecular/métodos , Sensibilidad y Especificidad

RESUMEN

Cat-transmitted sporotrichosis is caused by the emerging fungal pathogen Sporothrix brasiliensis and constitutes a significant public health issue that affects people living in resource-poor urban centers in Brazil. The lack of knowledge about transmission dynamics makes it difficult to propose public health policies to contain the advance of sporotrichosis. We describe the recent emergence of 1,176 cases of sporotrichosis in cats (2016 to 2021) in the metropolitan region of Recife, Brazil, leading to significant zoonotic transmission and an overwhelming occurrence of S. brasiliensis as the etiological agent. Most cases were from cats in the cities of Olinda (408/1,176; 34.70%), Jaboatão dos Guararapes (332/1,176; 28.23%), and Recife (237/1,176; 20.15%). Molecular typing using amplified fragment length polymorphism (EcoRI-GA/MseI-AG) revealed low polymorphic information content (PIC = 0.2499) and heterozygosity (H = 0.2928), typical of an outbreak scenario. Dendrogram and multivariate cluster analysis revealed that isolates from Pernambuco are closely related to Rio de Janeiro isolates. We report a substantial occurrence of MAT1-2 idiomorphs in the metropolitan region of Recife (0:60 ratio; χ2 = 60.000, P < 0.0001). The limited population differentiation and genetic diversity of the isolates from Pernambuco suggest a recent introduction, possibly via a founder effect, from the parental population in Rio de Janeiro. Our findings emphasize the critical importance of molecular surveillance of S. brasiliensis for outbreak response. A comprehensive one-health strategy is mandatory to control the spread of cat-transmitted sporotrichosis driven by S. brasiliensis, encompassing sanitary barriers, quick diagnosis, and treatment.

Asunto(s)

Enfermedades de los Gatos , Sporothrix , Esporotricosis , Esporotricosis/transmisión , Esporotricosis/microbiología , Esporotricosis/veterinaria , Esporotricosis/epidemiología , Gatos , Brasil/epidemiología , Sporothrix/genética , Sporothrix/aislamiento & purificación , Sporothrix/clasificación , Animales , Enfermedades de los Gatos/microbiología , Enfermedades de los Gatos/transmisión , Enfermedades de los Gatos/epidemiología , Tipificación Molecular , Zoonosis/transmisión , Zoonosis/microbiología , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Enfermedades Transmisibles Emergentes/transmisión , Enfermedades Transmisibles Emergentes/microbiología , Enfermedades Transmisibles Emergentes/epidemiología , Genotipo , Filogenia

RESUMEN

Sporotrichosis is recognized as the predominant subcutaneous mycosis in South America, attributed to pathogenic species within the Sporothrix genus. Notably, in Brazil, Sporothrix brasiliensis emerges as the principal species, exhibiting significant sapronotic, zoonotic and enzootic epidemic potential. Consequently, the discovery of novel therapeutic agents for the treatment of sporotrichosis is imperative. The present study is dedicated to the repositioning of pharmaceuticals for sporotrichosis therapy. To achieve this goal, we designed a pipeline with the following steps: (a) compilation and preparation of Sporothrix genome data; (b) identification of orthologous proteins among the species; (c) identification of homologous proteins in publicly available drug-target databases; (d) selection of Sporothrix essential targets using validated genes from Saccharomyces cerevisiae; (e) molecular modeling studies; and (f) experimental validation of selected candidates. Based on this approach, we were able to prioritize eight drugs for in vitro experimental validation. Among the evaluated compounds, everolimus and bifonazole demonstrated minimum inhibitory concentration (MIC) values of 0.5 µg/mL and 4.0 µg/mL, respectively. Subsequently, molecular docking studies suggest that bifonazole and everolimus may target specific proteins within S. brasiliensis- namely, sterol 14-α-demethylase and serine/threonine-protein kinase TOR, respectively. These findings shed light on the potential binding affinities and binding modes of bifonazole and everolimus with their probable targets, providing a preliminary understanding of the antifungal mechanism of action of these compounds. In conclusion, our research advances the understanding of the therapeutic potential of bifonazole and everolimus, supporting their further investigation as antifungal agents for sporotrichosis in prospective hit-to-lead and preclinical investigations.

Asunto(s)

Antifúngicos , Reposicionamiento de Medicamentos , Genoma Fúngico , Pruebas de Sensibilidad Microbiana , Sporothrix , Esporotricosis , Sporothrix/efectos de los fármacos , Sporothrix/genética , Antifúngicos/farmacología , Esporotricosis/microbiología , Esporotricosis/tratamiento farmacológico , Brasil , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/química , Simulación del Acoplamiento Molecular , Genómica , Humanos , Evaluación Preclínica de Medicamentos , Descubrimiento de Drogas , Biología Computacional

RESUMEN

A series of new metal complexes, [Cu(ITZ)2Cl2] ⋅ 5H2O (1), [Cu(NO3)2(ITZ)2] ⋅ 3H2O ⋅ C4H10O (2) and [Cu(ITZ)2)(PPh3)2]NO3 ⋅ 5H2O (3) were synthesized by a reaction of itraconazole (ITZ) with the respective copper salts under reflux. The metal complexes were characterized by elemental analyses, molar conductivity, 1H and 13C{1H} nuclear magnetic resonance, UV-Vis, infrared and EPR spectroscopies. The antifungal activity of these metal complexes was evaluated against the main sporotrichosis agents: Sporothrix brasiliensis, Sporothrix schenkii, and Sporothrix globosa. All three new compounds inhibited the growth of S. brasiliensis and S. schenckii at lower concentrations than the free azole, with complex 2 able to kill all species at 4 µM and induce more pronounced alterations in fungal cells. Complexes 2 and 3 exhibited higher selectivity and no mutagenic effect at the concentration that inhibited fungal growth and affected fungal cells. The strategy of coordinating itraconazole (ITZ) to copper was successful, since the corresponding metal complexes were more effective than the parent drug. Particularly, the promising antifungal activity of the Cu-ITZ complexes makes them potential candidates for the development of an alternative drug to treat mycoses.

Asunto(s)

Antifúngicos , Complejos de Coordinación , Cobre , Itraconazol , Pruebas de Sensibilidad Microbiana , Sporothrix , Cobre/química , Cobre/farmacología , Itraconazol/farmacología , Itraconazol/química , Sporothrix/efectos de los fármacos , Antifúngicos/farmacología , Antifúngicos/química , Antifúngicos/síntesis química , Complejos de Coordinación/farmacología , Complejos de Coordinación/química , Complejos de Coordinación/síntesis química , Relación Estructura-Actividad , Estructura Molecular , Relación Dosis-Respuesta a Droga

RESUMEN

We report the first case of Sporothrix brasiliensis in a dog at Santiago, Chile. From an initial biopsy obtained from the patient, a histopathological study was performed that oriented a fungal infection. The molecular diagnosis from the isolated colony confirmed the identity of the S. brasiliensis agent. Although the literature indicates itraconazole as the first treatment option, the patient was treated with terbinafine followed by itraconazole, without achieving a clinical cure. S. brasiliensis that spread rapidly in Latin America, hence highlight the importance of timely diagnosis that contribute to maintaining human and animal health. 2012 Elsevier Ltd. All rights reserved.

RESUMEN

Male cat, 2 years old, with a refractory infection by Sporothrix brasiliensis, presents a single nodular lesion in the left auricular pavilion. To confirm the diagnosis, cytology, fungal culture, antifungal susceptibility test, molecular analysis, and, to aid in the differential diagnosis, bacterial culture, antibiogram, and histopathology of the lesion were performed. In the absence of therapeutic success with conventional antifungals, photodynamic therapy (PDT) was introduced, demonstrating a satisfactory response in the sixth treatment session.

RESUMEN

The drugs available to treat sporotrichosis, an important yet neglected fungal infection, are limited. Some Sporothrix spp. strains present reduced susceptibility to these antifungals. Furthermore, some patients may not be indicated to use these drugs, while others may not respond to the therapy. The anthelmintic drug niclosamide is fungicidal against the Sporothrix brasiliensis type strain. This study aimed to evaluate whether niclosamide also has antifungal activity against Sporothrix globosa, Sporothrix schenckii and other S. brasiliensis strains with distinct genotypes and antifungal susceptibility status. Minimal inhibitory and fungicidal concentrations (MIC and MFC, respectively) were determined using the microdilution method according to the CLSI protocol. The checkerboard method was employed to evaluate niclosamide synergism with drugs used in sporotrichosis treatment. Metabolic activity of the strains under niclosamide treatment was evaluated using the resazurin dye. Niclosamide was active against all S. brasiliensis strains (n = 17), but it was ineffective (MIC > 20 µM) for some strains (n = 4) of other pathogenic Sporothrix species. Niclosamide MIC values for Sporothrix spp. were similar for mycelial and yeast-like forms of the strains (P = 0.6604). Niclosamide was fungicidal (MFC/MIC ratio ≤ 2) for most strains studied (89%). Niclosamide activity against S. brasiliensis is independent of the fungal genotype or non-wild-type phenotypes for amphotericin B, itraconazole, or terbinafine. These antifungal drugs presented indifferent interactions with niclosamide. Niclosamide has demonstrated potential for repurposing as a treatment for sporotrichosis, particularly in S. brasiliensis cases, instigating in vivo studies to validate the in vitro findings.

Asunto(s)

Antihelmínticos , Antifúngicos , Pruebas de Sensibilidad Microbiana , Niclosamida , Sporothrix , Sporothrix/efectos de los fármacos , Sporothrix/genética , Sporothrix/clasificación , Niclosamida/farmacología , Antifúngicos/farmacología , Antihelmínticos/farmacología , Esporotricosis/microbiología , Esporotricosis/tratamiento farmacológico , Genotipo , Humanos , Farmacorresistencia Fúngica , Sinergismo Farmacológico

RESUMEN

Sporotrichosis, the cutaneous mycosis most commonly reported in Latin America, is caused by the Sporothrix clinical clade species, including Sporothrix brasiliensis and Sporothrix schenckii sensu stricto. In Brazil, S. brasiliensis represents a vital health threat to humans and domestic animals due to its zoonotic transmission. Itraconazole, terbinafine, and amphotericin B are the most used antifungals for treating sporotrichosis. However, many strains of S. brasiliensis and S. schenckii have shown resistance to these agents, highlighting the importance of finding new therapeutic options. Here, we demonstrate that milteforan, a commercial veterinary product against dog leishmaniasis whose active principle is miltefosine, is a possible therapeutic alternative for the treatment of sporotrichosis, as observed by its fungicidal activity in vitro against different strains of S. brasiliensis and S. schenckii, and by its antifungal activity when used to treat infected epithelial cells and macrophages. Our results suggest milteforan as a possible alternative to treat feline sporotrichosis.

RESUMEN

Sporothrix brasiliensis is considered a highly virulent emerging pathogen that causes sporotrichosis in humans, mainly after zoonotic transmission from infected cats. The epidemic of this zoonosis that originated from Brazil has spread in the last decades, generating hyperendemic regions in Latin America. We present two cases of human sporotrichosis causes by S. brasiliensis in Buenos Aires, Argentina, with good clinical response to differing treatments after contact with sick cats. Using Short tandem repeat (STR) genotyping, the two S. brasiliensis cases appear to be introduced from Brazil and likely originate from the same source.

RESUMEN

BACKGROUND: Systemic sporotrichosis occurs when organs, other than subcutaneous tissues and lymph nodes, are infected. Interestingly, systemic sporotrichosis in apparently immunocompetent individuals is increasing in Brazil, but data on clinical manifestations and risk factors are scarce. Most of the existing data on such condition relate to people living with HIV. We aimed to study the risk factors associated with systemic sporotrichosis among HIV-negative and HIV-positive patients. METHODS: We performed a retrospective cross-sectional study with 80 patients from Brazil, diagnosed between 2014 and 2021. The association between disease classification, clinical presentation and risk factors were analysed by logistic regression. RESULTS: Of the 80 patients, 29 (36.3%) presented with systemic sporotrichosis. All HIV-positive patients developed the systemic form, with increased mortality (p = .002). Alcohol ingestion (p = .009) and diabetes (p = .010) were associated with systemic disease. Alcohol ingestion seemed to favour pulmonary infection (p = .013) and, diabetes favoured osteoarticular (p = .009) and ocular involvement (p = .033). The occurrence of fever (p = .001) and weight loss (p = .006) were significantly associated with systemic sporotrichosis, while meningeal involvement (p = .001) increased mortality risk. Nine (11.3%) patients died from sporotrichosis. The presence of fungal structures in the mycological examination of the patients' lesions were associated with the systemic form (p = .017) and death (p = .002). CONCLUSION: Our study points to the factors that drive systemic sporotrichosis other than HIV, such as alcohol ingestion and diabetes. Considering the high number of patients presenting severe sporotrichosis, the recognising these factors may contribute to timely diagnosis and proper treatment.

Asunto(s)

Diabetes Mellitus , Infecciones por VIH , Sporothrix , Esporotricosis , Humanos , Esporotricosis/microbiología , Brasil/epidemiología , Estudios Transversales , Estudios Retrospectivos

RESUMEN

Twenty-five years have passed since the initial observation of endemic zoonotic sporotrichosis in Rio de Janeiro, Brazil. Since then, this disease has spread throughout South America. Accompanying the emergence of this mycosis, some progress has been made, including the expansion of a research network in this field and higher visibility of sporotrichosis within government authorities and funding agencies. However, there are still some challenges to curbing the expansion of this disease in the coming years. These include the development of rapid and accurate diagnostic tests, new antifungal drugs, particularly for the treatment of extracutaneous manifestations of sporotrichosis, and more comprehensive care for cats with sporotrichosis. Including these actions in the sporotrichosis research agenda is required so as to change the development of this disease in the years to come.

RESUMEN

A esporotricose é uma zoonose micótica emergente e subcutânea, que afeta a pele, o sistema linfático e outros órgãos de humanos e animais. Assim como outras doenças infecciosas fúngicas, se torna ainda mais grave quando acomete pacientes imunossuprimidos. Essa infecção possui distribuição global e é endêmica em algumas regiões do Brasil e de outros países tropicais e subtropicais, sendo um problema de saúde pública importante em nosso país. A doença é causada por um complexo de pelo menos quatro espécies patogênicas, incluindo o Sporothrix brasiliensis (S. brasiliensis). A resposta imunológica contra estas espécies ainda não é completamente elucidada, mas estruturas como as vesículas extracelulares (VEs) poderiam transportar componentes importantes que podem contribuir na modulação e no controle desta importante infecção. Assim, o objetivo deste trabalho, é analisar a participação das VEs de células dendríticas (DCs) naive e VEs de DCs previamente primadas com leveduras de S. brasiliensis e primadas com VEs do fungo, na resposta imune contra a esporotricose experimental em modelos murinos. Para isso, as DCs obtidas da medula óssea de camundongos, foram cultivadas com leveduras de S. brasiliensis ou com VEs do fungo e posteriormente, VEs totais das DCs foram purificadas a partir de ultracentrifugação e analisadas quanto a sua participação na modulação da resposta imunológica. Essas VEs foram utilizadas em protocolo profilático em modelos murinos, previamente a infecção subcutânea experimental. Foi observado o diâmetro médio das lesões no decorrer de 35 dias de infecção e a carga fúngica da lesão na pele. Os resultados obtidos mostram que as VEs de DCs naive, e VEs de DCs previamente cultivadas com leveduras do fungo ou com VEs fúngicas, são capazes de modular a carga fúngica. Os grupos que receberam VEs de DCs de forma profilática, de modo geral apresentaram diminuição significativa da carga fúngica em relação ao grupo controle. Na análise comparativa apenas dos grupos que receberam a profilaxia, observa-se que o uso de VEs de DCs naive, resultam em uma carga fúngica maior que o uso de VEs de DCs previamente ativadas, e quando as DCs são ativadas com levedura, essa carga fúngica é a menor. Quando analisamos o perfil de citocinas na pele de camundongos tratados com as VEs previamente a infecção, observamos aumento de IFN-γ, TNF-α, IL-17 e IL-10 principalmente nos animais previamente tratados com VEs de DCs que foram ativadas com leveduras. Em relação às citocinas produzidas, podemos sugerir até o momento, uma resposta imunológica mista, mas que de alguma maneira, ainda não esclarecida, devem contribuir para melhor controle do processo infeccioso in vivo. Em relação a linfoproliferação, observa-se principalmente um aumento de linfócitos T CD4+ quando acrescentamos VEs de DCs que não foram previamente ativadas, mostrando uma ação de uma resposta mais inespecífica. Vale ressaltar que todos os protocolos profiláticos foram capazes de modular e minimizar o crescimento fúngico, quando comparados ao controle, ou seja, as VEs contribuíram com o controle da infecção e agiram a favor do hospedeiro, demonstrando um caráter protetivo

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Sporotrichosis is an emerging subcutaneous mycotic zoonosis that affects the skin, lymphatic system, and other organs of humans and animals, and like other infectious fungal diseases, it becomes even more serious when it affects immunosuppressed patients. This infection has a global distribution and is endemic in some regions of Brazil and other tropicals and subtropicals countries, being an important public health problem in our country. The disease is caused by a complex of at least four pathogenic species, including Sporothrix brasiliensis (S. brasiliensis). The immunological response against these species has not yet been completely elucidated, but structures such as extracellular vesicles (EVs) could carry important components that can contribute to the modulation and control of this important infection. Thus, the objective of this work is to analyze the participation of EVs from naïve dendritic cells (DCs) and EVs from DCs previously primed with S. brasiliensis yeast and primed with EVs from the fungus, in the immune response against experimental sporotrichosis in murine models. For this, DCs obtained from the bone marrow of mice were cultivated with S. brasiliensis yeast or EVs from the fungus, and subsequently, total EVs from the DCs were purified through ultracentrifugation and analyzed for their participation in modulating the immune response. These EVs were used in a prophylactic protocol in murine models, before experimental subcutaneous infection. The average diameter of the lesions over 35 days of infection and the fungal load of the lesion on the skin were observed. The results obtained show that EVs from naïve DCs, and EVs from DCs previously cultured with yeast or fungal EVs, are capable of modulating the fungal load. The groups that received EVs from DCs prophylactically generally showed a significant decrease in fungal load compared to the control group. In the comparative analysis of only the groups that received prophylaxis, it was observed that the use of EVs from naïve DCs results in a higher fungal load than the use of EVs from previously activated DCs, and when the DCs are activated with yeast, this load fungal is smaller. When we analyzed the cytokine profile in the skin of mice treated with EVs before infection, we observed an increase in IFN-γ, TNF-α, IL-17, and IL-10, mainly in animals previously treated with EVs from DCs that were activated with yeast. About the cytokines produced, we can so far suggest a mixed immunological response, but in some way, not yet clear, they should contribute to better control of the infectious process in vivo. About lymphoproliferation, an increase in CD4+ T lymphocytes is mainly observed when we add EVs from DCs that were not previously activated, showing a more nonspecific response. It is worth highlighting that all prophylactic protocols were able to modulate and minimize fungal growth, when compared to the control, that is, EVs contributed to the control of the infection and acted in favor of the host, demonstrating a protective character

Asunto(s)

Animales , Masculino , Ratones , Esporotricosis/patología , Células Dendríticas/clasificación , Vesículas Extracelulares/clasificación , Inmunidad , Enfermedades Transmisibles/tratamiento farmacológico , Hongos/aislamiento & purificación

RESUMEN

A esporotricose é uma zoonose micótica emergente e subcutânea, que afeta a pele, o sistema linfático e outros órgãos de humanos e animais. Assim como outras doenças infecciosas fúngicas, se torna ainda mais grave quando acomete pacientes imunossuprimidos. Essa infecção possui distribuição global e é endêmica em algumas regiões do Brasil e de outros países tropicais e subtropicais, sendo um problema de saúde pública importante em nosso país. A doença é causada por um complexo de pelo menos quatro espécies patogênicas, incluindo o Sporothrix brasiliensis (S. brasiliensis). A resposta imunológica contra estas espécies ainda não é completamente elucidada, mas estruturas como as vesículas extracelulares (VEs) poderiam transportar componentes importantes que podem contribuir na modulação e no controle desta importante infecção. Assim, o objetivo deste trabalho, é analisar a participação das VEs de células dendríticas (DCs) naive e VEs de DCs previamente primadas com leveduras de S. brasiliensis e primadas com VEs do fungo, na resposta imune contra a esporotricose experimental em modelos murinos. Para isso, as DCs obtidas da medula óssea de camundongos, foram cultivadas com leveduras de S. brasiliensis ou com VEs do fungo e posteriormente, VEs totais das DCs foram purificadas a partir de ultracentrifugação e analisadas quanto a sua participação na modulação da resposta imunológica. Essas VEs foram utilizadas em protocolo profilático em modelos murinos, previamente a infecção subcutânea experimental. Foi observado o diâmetro médio das lesões no decorrer de 35 dias de infecção e a carga fúngica da lesão na pele. Os resultados obtidos mostram que as VEs de DCs naive, e VEs de DCs previamente cultivadas com leveduras do fungo ou com VEs fúngicas, são capazes de modular a carga fúngica. Os grupos que receberam VEs de DCs de forma profilática, de modo geral apresentaram diminuição significativa da carga fúngica em relação ao grupo controle. Na análise comparativa apenas dos grupos que receberam a profilaxia, observa-se que o uso de VEs de DCs naive, resultam em uma carga fúngica maior que o uso de VEs de DCs previamente ativadas, e quando as DCs são ativadas com levedura, essa carga fúngica é a menor. Quando analisamos o perfil de citocinas na pele de camundongos tratados com as VEs previamente a infecção, observamos aumento de IFN-γ, TNF-α, IL-17 e IL-10 principalmente nos animais previamente tratados com VEs de DCs que foram ativadas com leveduras. Em relação às citocinas produzidas, podemos sugerir até o momento, uma resposta imunológica mista, mas que de alguma maneira, ainda não esclarecida, devem contribuir para melhor controle do processo infeccioso in vivo. Em relação a linfoproliferação, observa-se principalmente um aumento de linfócitos T CD4+ quando acrescentamos VEs de DCs que não foram previamente ativadas, mostrando uma ação de uma resposta mais inespecífica. Vale ressaltar que todos os protocolos profiláticos foram capazes de modular e minimizar o crescimento fúngico, quando comparados ao controle, ou seja, as VEs contribuíram com o controle da infecção e agiram a favor do hospedeiro, demonstrando um caráter protetivo

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Sporotrichosis is an emerging subcutaneous mycotic zoonosis that affects the skin, lymphatic system, and other organs of humans and animals, and like other infectious fungal diseases, it becomes even more serious when it affects immunosuppressed patients. This infection has a global distribution and is endemic in some regions of Brazil and other tropicals and subtropicals countries, being an important public health problem in our country. The disease is caused by a complex of at least four pathogenic species, including Sporothrix brasiliensis (S. brasiliensis). The immunological response against these species has not yet been completely elucidated, but structures such as extracellular vesicles (EVs) could carry important components that can contribute to the modulation and control of this important infection. Thus, the objective of this work is to analyze the participation of EVs from naïve dendritic cells (DCs) and EVs from DCs previously primed with S. brasiliensis yeast and primed with EVs from the fungus, in the immune response against experimental sporotrichosis in murine models. For this, DCs obtained from the bone marrow of mice were cultivated with S. brasiliensis yeast or EVs from the fungus, and subsequently, total EVs from the DCs were purified through ultracentrifugation and analyzed for their participation in modulating the immune response. These EVs were used in a prophylactic protocol in murine models, before experimental subcutaneous infection. The average diameter of the lesions over 35 days of infection and the fungal load of the lesion on the skin were observed. The results obtained show that EVs from naïve DCs, and EVs from DCs previously cultured with yeast or fungal EVs, are capable of modulating the fungal load. The groups that received EVs from DCs prophylactically generally showed a significant decrease in fungal load compared to the control group. In the comparative analysis of only the groups that received prophylaxis, it was observed that the use of EVs from naïve DCs results in a higher fungal load than the use of EVs from previously activated DCs, and when the DCs are activated with yeast, this load fungal is smaller. When we analyzed the cytokine profile in the skin of mice treated with EVs before infection, we observed an increase in IFN-γ, TNF-α, IL-17, and IL-10, mainly in animals previously treated with EVs from DCs that were activated with yeast. About the cytokines produced, we can so far suggest a mixed immunological response, but in some way, not yet clear, they should contribute to better control of the infectious process in vivo. About lymphoproliferation, an increase in CD4+ T lymphocytes is mainly observed when we add EVs from DCs that were not previously activated, showing a more nonspecific response. It is worth highlighting that all prophylactic protocols were able to modulate and minimize fungal growth, when compared to the control, that is, EVs contributed to the control of the infection and acted in favor of the host, demonstrating a protective character

Asunto(s)

Esporotricosis/patología , Células Dendríticas/clasificación , Heridas y Lesiones/clasificación , Enfermedades Transmisibles/complicaciones , Vesículas Extracelulares/clasificación

RESUMEN

Sporotrichosis is the most frequent subcutaneous or implantation mycosis in Latin America, and its transmission occurs as a result of traumatic inoculation into the skin by organic matter containing the thermodimorphic fungi of the genus Sporothrix. Although cutaneous forms are more common, another important site is the osteoarticular system, whose hematogenous involvement is commonly associated with disseminated forms, especially in people who have an immunosuppressive condition, such as HIV/AIDS, chronic steroid use, and alcohol abuse. We present two cases of osteoarticular sporotrichosis of the knee caused by Sporothrix brasiliensis and followed up at our institution, with different outcomes. In the cases presented here, aging, anatomical sites, comorbidities, subtherapeutic serum levels, low adherence to treatment, and late diagnosis for different reasons may explain the observed outcomes. Early diagnosis of Sporothrix infection is critical in preventing complications, including death. We also highlight the importance of multidisciplinary follow-up and adherence to treatment for a favorable outcome.

RESUMEN

Sporothrix brasiliensis is an emerging fungal pathogen causing cat-transmitted sporotrichosis, an epi-zoonosis affecting humans, cats and dogs in Brazil and now spreading to neighboring South American countries. Here, we report the first two autochthonous cases of cat-transmitted sporotrichosis in Paraguay. The first case was a four-year-old male cat showing several ulcerative lesions, nasal deformity and respiratory symptoms. The second case was a one-year-old male cat showing a single ulcerated lesion, respiratory symptoms and nasal deformity. Both cases were admitted to a veterinary clinic in Ciudad del Este, Paraguay. Isolates were recovered from swabs of the two cases. Using molecular methods, the isolates were identified as S. brasiliensis.

RESUMEN

Se describe el aislamiento de Sporothrix brasiliensis desde una biopsia de piel de un caso humano de esporotricosis linfocutánea, en la región de Valparaíso, Chile. Esta especie es la más virulenta del género y es de transmisión zoonótica, desde los gatos a los humanos. Hasta ahora, solo se había publicado un brote por esta especie en gatos domésticos y asilvestrados en el extremo sur de Chile, por lo que este aislamiento, en una mujer residente de un sector densamente poblado de la Región de Valparaíso, constituye una preocupación por su eventual diseminación hacia otros gatos y la población general.

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The isolation of Sporothrix brasiliensis from a skin biopsy of a human case of lymphocutaneous sporotrichosis in the region of Valparaíso, Chile is described. This species is the most virulent of the genus and is zoonotic in transmission from cats to humans. Until now, only one outbreak of this species has been published in domestic and feral cats in the extreme south of Chile, so this isolation in a woman residing in a densely populated sector of the fifth region is a concern for its eventual spread to other cats and the general population.

Asunto(s)

Humanos , Animales , Gatos , Esporotricosis/microbiología , Sporothrix/aislamiento & purificación , Esporotricosis/transmisión , Sporothrix/genética , Zoonosis , Chile