Responsible ownership and health education can reduce the time of sporotrichosis treatment in domestic cats.

Sporotrichosis is a widespread fungal infection that affects skin and subcutaneous tissues in humans and animals. In cats, it is displayed as nodules, ulcers and lesions on the nasal and respiratory mucosa. Antifungal treatment of cats is crucial but many cases are difficult, thus resulting in discontinue of the treatment, with disastrous consequences for the animal, encouraging contamination of the environment, other animals and people. The effects of responsible ownership education and health education for owners of cats with feline sporotrichosis as well as the interval between veterinary consultations on treatment outcomes for three groups of owners and their pet cats were evaluated in this study. The responsible ownership education and health education strategies consisted in videos in easy and accessible language for people with any level of education and were presented during consultations for two of the three groups included. The time between appointments was two weeks for two of the groups, and four weeks for one of the groups. The median of treatment time for the group without educational activities was 138 days, while for the other two groups it was 77.5 days and 86 days. It was found a significative reduction in the treatment time in the groups exposed to Responsible ownership education videos. There was no contamination of those responsible for home treatment, and the interval between monthly appointments did not impact on cure or death rates compared to the interval between fortnightly appointments. All these results can be applied to feline sporotrichoses treatment protocols increasing the owners treatment adherence and reducing either, the treatment discontinuation and the treatment costs and helps to control zoonotic sporotrichosis. The importance of attractive and comprehensible educational strategies as part of the feline sporotrichosis treatment protocol for the promotion of one health was highlighted.

[Public health risk and prevention and control of sporotrichosis].

Sporotrichosis, a fungal infection caused by Sporothrix species, can greatly lead to chronic inflammation of the skin, mucosa, and lymphatic vessels and disseminate systemically sometimes, even threatening life. It is known that Sporothrix is distributed worldwide, while in China, most of the cases were reported in northeast China and parts of south China. Sporothrix globosa is the main source of infection, and other regions may lack relevant awareness and attention to the disease, making it a public health challenge in China. Thus, it is important to understand its epidemiology and public health risks to prevent and control the disease properly.

Mathematical model of the dynamics of transmission and control of sporotrichosis in domestic cats.

Sporotrichosis is a subcutaneous mycosis with a global distribution, also known as "rose gardener's disease". Brazil is experiencing a rapid spread of the zoonotic transmission of of Sporothrix brasiliensis, the main etiological agent of this disease in this country, affecting domestic felines. Cost-effective interventions need to be developed to control this emergent public health problem. To allow for the comparison of alternative control strategies, we propose in this paper, a mathematical model representing the transmission of S. brasiliensis among cats, stratified by age and sex. Analytical properties of the model are derived and simulations show possible strategies for reducing the endemic levels of the disease in the cat population, with a positive impact on human health. The scenarios included mass treatment of infected cats and mass implementation of contact reduction practices, such as neutering. The results indicate that mass treatment can reduce substantially the disease prevalence, and this effect is potentialized when combined with neutering or other contact-reduction interventions. On the other hand, contact-reduction methods alone are not sufficient to reduce prevalence.

A Sporothrix spp. enolase derived multi-epitope vaccine confers protective response in BALB/c mice challenged with Sporothrix brasiliensis.

Sporotrichosis is a cosmopolitan mycosis caused by pathogenic species of Sporothrix genus, that in Brazil is often acquired by zoonotic transmission involved infected cats with S. brasiliensis. Previous studies showed that the Sporothrix spp. recombinant enolase (rSsEno), a multifunctional protein with immunogenic properties, could be a promising target for vaccination against sporotrichosis in cats. Nevertheless, the considerable sequence identity (62%) of SsEno with its feline counterpart is a great concern. Here, we report the identification in silico, chemical synthesis and biological validation of six peptides of SsEno with low sequence identity to its cat orthologue. All synthesized peptides exhibit B-cell epitopes on the molecular surface of SsEno and proved to be highly reactive with the serum of infected mice with S. brasiliensis and sera of cats with sporotrichosis. Interestingly, our study revealed that anti-peptide sera did not react with the recombinant enolase from Felis catus (cats, rFcEno), thus, may not trigger autoimmune response in these felines if used as a vaccine antigen. The immunization with peptide mixture (PeptMix) formulated with Freund adjuvant (FA), induced high levels of antigen-specific IgG, IgG1 and IgG2b antibodies that conferred protection upon passive transference in infected BALB/c mice with S. brasiliensis. We also observed, that the FA+PeptMix formulation induced a Th1/Th2/Th17 cytokine profile ex vivo, associated with protecting effect against the experimental sporotrichosis. Our results suggest that the six SsEno-derived peptides here evaluated, could be used as safe antigens for the development of vaccine strategies against feline sporotrichosis, whether prophylactic or therapeutic.

Comorbidity profile of patients with concurrent diagnoses of asthma and COPD in Germany.

The aim of this study was to estimate the prevalence of concurrent diagnoses of asthma/COPD and examine its full spectrum of comorbid disorders in Germany. We used nationwide outpatient claims data comprising diagnoses of all statutory health insurees (40+ years) in 2017 (N = 40,477,745). The ICD-10 codes J44 (COPD) and J45 (asthma) were used to identify patients. The odds of 1,060 comorbid disorders were examined in a case-control study design. Of all insurees, 4,632,295 (11%) were diagnosed with either asthma or COPD. Of them, 43% had asthma only, 44% COPD only and 13% both diseases. The prevalence of concurrent asthma/COPD was 1.5% with a slightly higher estimate among females than males (1.6% vs. 1.4%) and constant increase by age in both sexes. Comorbid disorders were very common among these patients. 31 disorders were associated with a strong effect size (odds ratio > 10), including other respiratory diseases, but also bacterial (e.g., mycobacteria, including tuberculosis) and fungal infections (e.g., sporotrichosis and aspergillosis). Patients with concurrent asthma/COPD suffer from comorbid disorders involving various body systems, which points to the need of a multidisciplinary care approach. Regular screening for common comorbid disorders may result in better clinical course and prognosis as well as improvement of patients' quality of life.

Nota técnica 09 DVE/DVZ/COVISA/2020: vigilância e manejo clínico da esporotricose humana no município de São Paulo / Technical report 09 DVE / DVZ / COVISA / 2020: surveillance and clinical management of human sporotrichosis in the city of São Paulo

A Esporotricose é uma infecção fúngica de implantação, subaguda ou crônica, causada por fungo do complexo Sporothrix schenkii. No Brasil, a espécie mais frequente é S.brasiliensis. No município de São Paulo, em 2011, foram identificados os primeiros casos de esporotricose em felinos e em humanos, na região de Itaquera, na Coordenadoria Regional de Saúde Leste. A partir de 2018, houve um aumento significativo do número de casos em felinos, acompanhado do aumento de casos humanos. Esta Nota técnica, elaborada pela Coordenadoria de Vigilância em Saúde de São Paulo, tem como objetivo orientar a vigilância e o manejo clínico da esporotricose humana no município de São Paulo. O documento aborda a situação epidemiológica da esporotricose no município de São Paulo, o agente etiológico, as formas de transmissão, as manifestações clínicas, os diagnósticos diferenciais, o diagnóstico laboratorial, o tratamento, o prognóstico e as definições de casos suspeito e confirmado, além dos fluxos de atendimento e vigilância epidemiológica de esporotricose humana na cidade e da orientação para coleta e armazenamento de amostras para envio ao laboratório.

The threat of emerging and re-emerging pathogenic Sporothrix species.

Sporotrichosis is a neglected subcutaneous mycosis of humans and animals acquired by traumatic inoculation of soil and plant material (classical route) contaminated with infectious propagules of the pathogen or being bitten/scratched by infected cats (alternative route). Within a genus composed of 53 species displaying an essentially environmental core, there are only a few members which have considerable impacts on human or animal health. Infections are typically caused by S. brasiliensis, S. schenckii or S. globosa. Rare mammal pathogens include members of the S. pallida and S. stenocereus complexes. To illustrate the tremendous impact of emerging zoonotic sporotrichosis on public health, we discuss the main features of the expanding epidemics driven by S. brasiliensis in cats and humans. The cat entry in the transmission chain of sporotrichosis, causing epizooties (cat-cat) or zoonosis (cat-human), has contributed to the definition of new paradigms in Sporothrix transmission, reaching epidemic levels, making the disease a serious public health problem. Indeed, S. brasiliensis infection in humans and animals is likely to become even more important in the future, with projections of its expansion in biogeographic domains and host range, as well as greater virulence in mammals. Therefore, lessons from a long-standing outbreak in the state of Rio de Janeiro about the source and distribution of the etiological agents among outbreak areas can be used to create better control and prevention plans and increase awareness of sporotrichosis as a serious emerging zoonotic disease.

Nota técnica 09 - vigilância e manejo clínico da esporotricose humana no município de São Paulo / Technical note 09 - surveillance and clinical management of human sporotrichosis in the city of São Paulo

Nota técnica elaborada para nortear a vigilância e manejo clínico da esporotricose humana no município de São Paulo. O documento aborda a situação da esporotricose no município de São Paulo, agente etiológico, transmissão, manifestações clínicas da esporotricose humana, diagnóstico diferencial, diagnóstico laboratorial, tratamento, prognóstico, unidades de referencia, vigilância epidemiológica e fluxos de atendimento de casos humanos. Apresenta ainda orientações para coleta, armazenamento e ficha de encaminhamento de amostras para diagnóstico laboratorial, ficha para notificação, ficha de encaminhamento de caso suspeito e fluxograma de atendimento de casos humanos.

Immunization with recombinant enolase of Sporothrix spp. (rSsEno) confers effective protection against sporotrichosis in mice.

In recent years, research has focused on the immunoreactive components of the Sporothrix schenckii cell wall that can be relevant targets for preventive and therapeutic vaccines against sporotrichosis, an emergent worldwide mycosis. In a previous study, we identified a 47-kDa enolase as an immunodominant antigen in mice vaccinated with an adjuvanted mixture of S. schenckii cell wall proteins. Here, we sought to assess the protective potential of a Sporothrix spp. recombinant enolase (rSsEno) formulated with or without the adjuvant Montanide Pet-GelA (PGA) against the S. brasiliensis infection in mice. Mice that were immunized with rSsEno plus PGA showed increased antibody titters against rSsEno and increased median survival time when challenged with S. brasiliensis as compared with mice that had not been immunized or that were immunized with rSsEno alone. Immunization with rSsEno plus PGA induced a predominantly T-helper 1 cytokine pattern after in vitro stimulation of splenic cells with rSsEno: elevated levels of IFN-γ and IL-2, as well as of other cytokines involved in host defense against sporotrichosis, such as TNF-alpha, IL-6, and IL-4. Furthermore, we show for the first time the presence of enolase in the cell wall of both S. schenckii and S. brasiliensis. As a whole, our results suggest that enolase could be used as a potential antigenic target for vaccinal purposes against sporotrichosis.

Prophylactic and therapeutic vaccines against sporotrichosis. Feasibility and prospects.

Sporotrichosis is an emergent subcutaneous mycosis of humans and some animals caused by dimorphic fungi of the genus Sporothrix. The disease occurs worldwide but is endemic or hyperendemic in tropical and subtropical areas. The epidemiology of the disease is changing dramatically, and it is now considered an important zoonosis with high morbidity rates, principally in Brazil, and an opportunistic infection in immunocompromised patients. Due to the limited options currently available to treat invasive fungal infections, including sporotrichosis, and the emergence of drug resistance and toxicity, the development of anti-Sporothrix vaccines has become an area of great interest. This work provides a brief analysis of the feasibility of the development of prophylactic and therapeutic vaccines against sporotrichosis, the main advances achieved to date, future challenges and prospects.

Situação atual da Vigilância da Esporotricose

Situação atual da Vigilância da Esporotricose

An immunoproteomic approach revealing peptides from Sporothrix brasiliensis that induce a cellular immune response in subcutaneous sporotrichosis.

Sporothrix brasiliensis is the most virulent fungus of the Sporothrix complex and is the main species recovered in the sporotrichosis zoonotic hyperendemic area in Rio de Janeiro. A vaccine against S. brasiliensis could improve the current sporotrichosis situation. Here, we show 3 peptides from S. brasiliensis immunogenic proteins that have a higher likelihood for engaging MHC-class II molecules. We investigated the efficiency of the peptides as vaccines for preventing subcutaneous sporotrichosis. In this study, we observed a decrease in lesion diameters in peptide-immunized mice, showing that the peptides could induce a protective immune response against subcutaneous sporotrichosis. ZR8 peptide is from the GP70 protein, the main antigen of the Sporothrix complex, and was the best potential vaccine candidate by increasing CD4+ T cells and higher levels of IFN-γ, IL-17A and IL-1ß characterizing a strong cellular immune response. This immune environment induced a higher number of neutrophils in lesions that are associated with fungus clearance. These results indicated that the ZR8 peptide induces a protective immune response against subcutaneous sporotrichosis and is a vaccine candidate against S. brasiliensis infection.

Caracterização e avaliação da capacidade protetora dos peptí­deos imunogênicos de Sporothrix brasiliensis / Characterization and evaluation of the protective capacity of the immunogenic peptides of Sporothrix brasiliensis

A esporotricose é uma doença crônica que envolve o tecido subcutâneo afetando seres humanos e animais, causada pelo fungo termodimórfico Sporothrix spp.. A esporotricose é endêmica na América latina, principalmente no Brasil que teve o maior surto zoonótico já registrado, ocorrendo na cidade do Rio de Janeiro. A espécie Sporothrix brasiliensis é a mais diagnosticada no surto e a mais virulenta entre as especies de Sporothrix spp., causando formas mais graves da doença. A esporotricose em gatos é endêmica, fatal e um dos principais fatores pelo alto número de casos no Rio de Janeiro. O tratamento é longo e não vem sendo o suficiente para conter o número de casos da doença. Uma vacina contra a esporotricose poderia mudar esse paradigma no Brasil. O presente trabalho obteve o proteoma da cepa S. brasiliensis 5110 por meio de uma eletroforese 2D, e caracterizou e identificou as possíveis proteínas imunogênicas do fungo por espectrometria de massa. Por meio de programas de predição, foi avaliado e sintetizado 7 sequências de aminoácidos,das proteínas identificadas com maiores chances de se acoplar a molécula MHC de classe II. Apenas 3 foram capazes de induzir proliferação in vitro, os peptídeos ZR3, ZR4 e ZR8, que foram utilizados como vacina na esporotricose subcutânea e avaliados sua eficácia por meio da carga fúngica, diâmetro das lesões, perfil celular e níveis de citocinas. Neste trabalho concluímos que o peptídeo ZR8 foi o melhor candidato à vacina na esporotricose, pois foi capaz de diminuir o diâmetro das lesões, aumentar os níveis de citocinas protetoras (IFN-γ, IL-17A e IL-1ß) e aumentar o número de células TCD4+ e CD3-/CD19+, sendo assim induzindo uma resposta imunológica protetora na esporotricose subcutânea

Sporotrichosis is a chronic disease, which involves the subcutaneous tissue affecting humans and animals caused by the thermodymorphic fungus Sporothrix spp. Sporotrichosis is endemic in Latin America, mainly in Brazil that had the largest zoonotic outbreak ever recorded, occurring in the city of Rio de Janeiro. The Sporothrix brasiliensis is the species more diagnosed in the outbreak and most virulent, causing severe forms of the disease. Sporotrichosis in cats is endemic, fatal and the main factors due to the high number of cases of the disease in Rio de Janeiro. The treatment is long, and has not been enough to contain the number of cases of sporotrichosis. A vaccine against sporotrichosis could change this paradigm in Brazil. The present work obtained the proteome of S. brasiliensis 5110 strain by 2D electrophoresis, and characterized and identified possible immunogenic proteins by mass spectrometry. By prediction programs were evaluated and synthesized 7 peptide sequence from antigenic proteins that have the highest chances of coupling to the MHC class II molecule. From these 7 peptides only 3 were able to induce proliferation in vitro, called ZR3, ZR4 and ZR8 peptides, that were used as a vaccine in subcutaneous sporotrichosis and evaluated their efficacy through fungal load, lesion diameter, cell profile and cytokine levels. We conclude that ZR8 peptide was the best candidate for sporotrichosis vaccine, since it was able to decrease the lesion diameter, increase the levels of protective cytokines (IFN-γ, IL-17A and IL-1ß) and increase the number of CD4+ T cells and CD3-/CD19+ inducing a protective immune response in subcutaneous sporotrichosis

Comparative efficacy and toxicity of two vaccine candidates against Sporothrix schenckii using either Montanide™ Pet Gel A or aluminum hydroxide adjuvants in mice.

Sporotrichosis is an important zoonosis in Brazil and the most frequent subcutaneous mycosis in Latin America, caused by different Sporothrix species. Currently, there is no effective vaccine available to prevent this disease. In this study, the efficacy and toxicity of the adjuvant Montanide™ Pet Gel A (PGA) formulated with S. schenckii cell wall proteins (ssCWP) was evaluated and compared with that of aluminum hydroxide (AH). Balb/c mice received two subcutaneous doses (1st and 14th days) of either the unadjuvanted or adjuvanted vaccine candidates. On the 21st day, anti-ssCWP antibody levels (ELISA), the phagocytic index, as well as the ex vivo release of IFN-γ, IL-4, and IL-17 by splenocytes and IL-12 by peritoneal macrophages were assessed. Cytotoxicity of the vaccine formulations was evaluated in vitro and by histopathological analysis of the inoculation site. Both adjuvanted vaccine formulations increased anti-ssCWP IgG, IgG1, IgG2a, and IgG3 levels, although IgG2a levels were higher in response to PGA+CWP100, probably contributing to the increase in S. schenckii yeast phagocytosis by macrophages in the opsonophagocytosis assay when using serum from PGA+CWP100-immunized mice. Immunization with AH+CWP100 led to a mixed Th1/Th2/Th17 ex vivo cytokine release profile, while PGA+CWP100 stimulated a preferential Th1/Th2 profile. Moreover, PGA+CWP100 was less cytotoxic in vitro, caused less local toxicity and led to a similar reduction in fungal load in the liver and spleen of S. schenckii- or S. brasiliensis-challenged mice as compared with AH+CWP100. These results suggest that PGA may be an effective and safe adjuvant for a future sporotrichosis vaccine.

Esporotricose: perguntas e respostas

Notícias, perguntas e respostas a respeito da esporotricose - doença fúngica de felinos com transmissão para humanos.

Anti-inflammatory activity of Vismia guianensis (Aubl.) Pers. extracts and antifungal activity against Sporothrix schenckii.

ETHNOPHARMACOLOGICAL RELEVANCE: Vismia guianensis (Aubl.) Pers. is traditionally used in North and Northeast of Brazil for the treatment of dermatomycoses. Since the strategy associating immunomodulators with antifungal drugs seems to be promissory to improve the treatment efficacy in fungal infections, we aimed to investigate the antifungal activity of V. guianensis ethanolic extract of leaves (VGL) and bark (VGB) against Sporothrix schenckii ATCC 16345 and their antinflammatory activities. MATERIAL AND METHODS: The extracts were analyzed by HPLC-DAD-IT MS/MS for in situ identification of major compounds. Antifungal activity was evaluated in vitro (microdilution test) and in vivo using a murine model of S. schenckii infection. The production of TNF-α, IFN-γ, IL-4, IL-10 and IL-12 by measured by ELISA, as well as measured the production and inhibition of the NO after treatment with the plant extracts or itraconazole (ITR). RESULTS: Two O-glucosyl-flavonoids and 16 prenylated benzophenone derivatives already described for Vismia were detected. Both VGL and VGB showed significant antifungal activity either in in vitro assay of microdilution (MIC=3.9µg/mL) and in vivo model of infection with reduction of S. schenckii load in spleen. It was also observed a predominance of reduction in the production of NO and the proinflammatory cytokines evaluated except TNFα, but with stimulation of IL-10, as evidence of a potential anti-inflammatory effect associated. CONCLUSION: The results showed that both VGL and VGB have a significant antifungal against S. schenckii and an anti-inflammatory activity. These results can support the use of these extracts for alternative treatment of sporotrichosis.

Is epidemic of sporotrichosis in Abancay, Peru, caused by zoonotic transmission of Sporothrix? / ¿Es la epidemia de esporotricosis en Abancay, Perú, debida a la transmisión zoonótica de Sporothrix?

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Effects of essential oils of Rosmarinus officinalis Linn. and Origanum vulgare Linn. from different origins on Sporothrix brasiliensis and Sporothrix schenckii complex / Efeitos dos óleos essenciais de Rosmarinus officinalis Linn. e Origanum vulgare Linn. de diferentes origens em Sporothrix brasiliensis e complexo Sporothrix schenckii

Rosmarinus officinalis L. (rosemary) and Origanum vulgare L. (oregano) are known to have antimicrobial properties, but studies on sporotrichosis are scarce. This study aimed to evaluate the anti-Sporothrix spp. activity of essential oils from commercial products and oils extracted from aerial parts of these plants and analyze their chemical constituents. S. schenckii complex and S. brasiliensis (n: 25) isolated from humans, cats, dogs, and environmental soil were tested through M27-A3 guidelines of CLSI with modification for phytotherapics. The essential oils of R. officinalis L. were similar for MIC50 and MFC50 ≤2.25mg/mL for extracted oil; and 4.5mg/mL and 9mg/mL, respectively, for commercial oil. Both products showed MIC90 of 18mg/mL and MFC90 of 36mg/mL. In O. vulgare L., the extracted oil had better activity with MIC50 and MFC50 ≤2.25mg/mL, and MIC90 and MFC90 of 4.5mg/mL, whereas the commercial oil showed MIC50 and MFC50 of 9mg/mL and MIC90 18mg/mL, respectively, and MFC90 of 36mg/mL. Through gas chromatography (CG/FID), thymol and α-terpinene were majority for extracted oil of O. vulgare L., and carvacrol and γ-terpinene made up the majority of the commercial oil. Both essential oils of R. officinalis L. showed 1,8-cineole and α-pinene as major. The fungal isolates were susceptible to all tested essential oils, including in itraconazole-resistant S. brasiliensis isolates. The extracted and commercial oils of the plants presented in vitro anti-Sporothrix spp. activity, and they are promising for treatment of sporotrichosis, including in cases refractory to itraconazole. More studies should be performed about toxicity and in vivo efficacy for its safe use.(AU)

Rosmarinus officinalis L. (alecrim) e Origanum vulgare L. (orégano) são conhecidos pelas propriedades antimicrobianas, entretanto seus estudos na esporotricose são escassos. Este trabalho objetivou avaliar a atividade anti-Sporothrix spp. de óleos extraídos e comerciais dessas plantas e analisar seus constituintes químicos. Isolados do complexo S. schenckii e S. brasiliensis (n: 25) de humanos, gatos, cães e solo, foram testados pela diretriz M27-A3 do CLSI com modificações para fitoterápicos. Os óleos de R. officinalis L. foram similares com CIM50 e CFM50 ≤2.25mg/mL para extraído; e 4.5mg/mL e 9mg/mL, respectivamente, para comercial. Ambos os produtos demonstraram CIM90 de 18mg/mL e CFM90 de 36mg/mL. Em O. vulgare L., o óleo extraído apresentou melhor atividade com CIM50 e CFM50≤2.25mg/mL e CIM90 e CFM90 de 4.5mg/mL, ao passo que o óleo comercial mostrou CIM50 e CFM50 de 9mg/mL; e CIM90 de 18mg/mL e CFM90 de 36mg/mL. Por meio da cromatografia gasosa (CG/FID), timol e α-terpineno foram majoritários para o óleo extraído de O. vulgare L., e carvacrol e γ-terpineno para o comercial. Ambos os óleos de R. officinalis L. apresentaram 1,8-cineol e α-pineno como prevalentes. Os isolados foram sensíveis a todos os óleos essenciais testados, inclusive S. brasiliensis, resistentes ao itraconazol. Os óleos extraídos e comerciais de R. officinalis L. e O. vulgare L. apresentaram atividade anti-Sporothrix spp. in vitro e são promissores para o tratamento da esporotricose, inclusive em casos refratários ao itraconazol. Mais estudos devem ser realizados sobre toxicidade e eficácia in vivo para seu uso seguro.(AU)

A cell wall protein-based vaccine candidate induce protective immune response against Sporothrix schenckii infection.

Sporotrichosis is a subcutaneous mycosis caused by several closely related thermo-dimorphic fungi of the Sporothrix schenckii species complex, affecting humans and other mammals. In the last few years, new strategies have been proposed for controlling sporotrichosis owning to concerns about its growing incidence in humans, cats, and dogs in Brazil, as well as the toxicity and limited efficacy of conventional antifungal drugs. In this study, we assessed the immunogenicity and protective properties of two aluminum hydroxide (AH)-adsorbed S. schenckii cell wall protein (ssCWP)-based vaccine formulations in a mouse model of systemic S. schenckii infection. Fractioning by SDS-PAGE revealed nine protein bands, two of which were functionally characterized: a 44kDa peptide hydrolase and a 47kDa enolase, which was predicted to be an adhesin. Sera from immunized mice recognized the 47kDa enolase and another unidentified 71kDa protein, whereas serum from S. schenckii-infected mice recognized both these proteins plus another unidentified 9.4kDa protein. Furthermore, opsonization with the anti-ssCWP sera led to markedly increased phagocytosis and was able to strongly inhibit the fungus' adhesion to fibroblasts. Immunization with the higher-dose AH-adjuvanted formulation led to increased ex vivo release of IL-12, IFN-γ, IL-4, and IL-17, whereas only IL-12 and IFN-γ were induced by the higher-dose non-adjuvanted formulation. Lastly, passive transference of the higher-dose AH-adjuvanted formulation's anti-ssCWP serum was able to afford in vivo protection in a subsequent challenge with S. schenckii, becoming a viable vaccine candidate for further testing.