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Sporotrichosis is a mycotic disease caused by Sporothrix spp., whose zoonotic transmission by sick cats is the main infection route in Brazil. The aim of the current study is to report a human sporotrichosis outbreak caused by zoonotic transmission from a feline, with emphasis on the importance of making differential diagnosis and of using personal protective equipment. A hospital team member presented injury in the arm after having handled a cat that had been hospitalized for being hit by a car. The animal presented skin lacerations, myiasis, and full tibial fracture - there were no other signs of skin lesions. Clinical samples were collected from both the human and the suspected cat, for mycological culture; results have shown Sporothrix sp. growth. A search was conducted to identify other hospital team members who also had contact with the animal. Other six individuals also had suspected lesions in their arms, hands and ocular area; they were all subjected to sample collection. Mycological results have also confirmed Sporothrix spp.; sequencing analysis has shown that all seven humans were infected with Sporothrix brasiliensis. Since Southern Brazil is endemic of this disease, it is worth emphasizing the importance of taking into consideration zoonotic risks at the time to provide emergency care to stray animals, mainly felines, as well as of using Personal Protective Equipment while handling them - regardless of whether they present, or not, typical clinical symptoms or history of the disease, given the potential zoonotic risk posed by Sporothrix brasiliensis.

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From drug repurposing studies, this work aimed to evaluate the activity of different pyrazinoic acid (POA) derivatives against Sporothrix brasiliensis. The POA esters were prepared and characterized as previously reported by classical esterification reactions, with good to excellent yields. Sporothrix brasiliensis isolates from cats (n=6) and standard strains of S. brasiliensis and S. schenckii were used to assess the antifungal activity of the POA derivatives through broth microdilution assay (CLSI M38-A2). Among the tested compounds, molecules 3 and 4 showed fungistatic and fungicidal activities against all Sporothrix spp. strains, and the obtained MIC and MFC values ranged from 2.12 to 4.24 mg/mL and from 1.29 to 5.15 mg/mL, respectively. Compound 2 and 5 were active as in vitro inhibitors of fungal growth, but showed weak fungicidal activity, while molecules 1 and POA itself were inactive. The results suggest the activity of POA derivatives against Sporothrix spp. may be dependent on the lipophilicity. In addition, the antifungal susceptibility of the isolates to itraconazole was performed, showing that two Sporothrix isolates from cats were itraconazole-resistant. Compounds 3 and 4 were also active against these itraconazole-resistant isolates, indicating a possible alternative route to the standard mode of action of itraconazole.

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PURPOSE: Motivated by increasing reports of antifungal resistance in human and animal sporotrichosis, this study evaluated the chemical composition, cytotoxicity and anti-Sporothrix brasiliensis activity of extracts of marjoram (Origanum majorana) and rosemary (Rosmarinus officinalis). METHODOLOGY: Ten (INF10) and 60 min (INF60) infusions, a decoction and a hydroalcoholic extract (HAE, 70 %) were prepared from both plants (10 % w/v). The extract composition was analysed by liquid chromatography/mass spectrometry and the cytotoxicity was evaluated using a colorimetric assay in canine and feline kidney cells. Using a broth microdilution assay (CLSI M38-A2) adapted to the extracts, 30 Sporothrix brasiliensis isolates from dogs, cats and humans, and one Sporothrix schenckii were tested.Results/Key findings. The predominant phenolic compounds found in all extracts were 4-hydroxybenzoic acid, caffeic acid and chlorogenic acid. Luteolin was also one of the predominant compounds, but only in the HAE of marjoram. Extracts of marjoram maintained cell viability in concentrations up to 2.5 mg ml-1 for the feline cell line and up to 10 mg ml-1 for the canine cell line, whereas in rosemary, the cell viability for both kidney lines was maintained with concentrations up to 5 mg ml-1. The activity of rosemary extracts was low or absent. Among the marjoram extracts, HAE was highlighted and had fungistatic activity against Sporothrix brasiliensis (MIC5040 mg ml-1), including in all itraconazole-resistant isolates. S. schenckiisensu stricto was sensitive to marjoram extracts (MIC/MFC ≤5 mg ml-1), with the exception of INF10. CONCLUSION: These findings support the potential usefulness of the HAE of marjoram in the treatment of sporotrichosis.

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The study aimed to evaluate the anti-Sporothrix sp. activity of the essential oil of Origanum majorana Linn. (marjoram), its chemical analysis, and its cytotoxic activity. A total of 18 fungal isolates of Sporothrix brasiliensis (n: 17) from humans, dogs and cats, and a standard strain of Sporothrix schenckii (n: 1) were tested using the broth microdilution technique (Clinical and Laboratory Standard Institute - CLSI M27-A3) and the results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC). The MIC50 and MIC90 of itraconazole against S. brasiliensis were 2 µg/mL and 8 µg/mL, respectively, and the MFC50 and MFC90 were 2 µg/mL and >16 µg/mL, respectively, with three S. brasiliensis isolates resistant to antifungal. S. schenckii was sensitive at MIC of 1 µg/mL and MFC of 8 µg/mL. For the oil of O. majorana L., all isolates were susceptible to MIC of 2.25-9 mg/mL and MFC of 2.25-18 mg/mL. The MIC50 and MIC90 were 2.25 mg/mL and 4.5 mg/mL, respectively, and the MFC50/90 values were twice more than the MIC. Twenty-two compounds were identified by gas chromatography with a flame ionization detector (CG-FID) and 1,8-cineole and 4-terpineol were the majority. Through the colorimetric (MTT) assay, the toxicity was observed in 70-80% of VERO cells between 0.078 and 5 mg/mL. For the first time, the study demonstrated the satisfactory in vitro anti-Sporothrix sp. activity of marjoram oil and further studies are needed to ensure its safe and effective use.(AU)

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ABSTRACT Cases of sporotrichosis in humans and animals without satisfactory clinical response have increased, a warning sign of strains resistant to conventional antifungal agents. The urgent search for alternative therapies was an incentive for research on medicinal plants with anti-Sporothrix spp. properties. A bibliographic survey was performed based on scientific papers about in vitro and in vivo antifungal activity of essential oils and extracts of plants in differents solvents against the fungal of the Sporothrix schenckii complex. The study methodology consisted of a literature review in Google Scholar, Science Direct, Pubmed, Bireme and Springer link with papers from 1986 to 2015. We found 141 species of plants that were investigated, of which 100 species were concentrated in 39 botanical families that had confirmed anti-Sporothrix activity. Combretaceae, Asteraceae and Lamiaceae represented the botanical families with the greatest number of plants species with antifungal potential, using different methodologies. However, there are few studies with medicinal plants in experimental infection in animals that prove their activity in the treatment of sporotrichosis. It reinforces the need for further research related to standardization of in vitro methodologies and in vivo studies related to safety and to toxicity potential of these plants with anti-Sporothrix spp. activity.

RESUMO Casos de esporotricose em humanos e animais sem resposta clínica satisfatória têm aumentado, sinal de alarme para o surgimento de cepas resistentes aos antifúngicos convencionais. A urgente busca por alternativas terapêuticas tem incentivado as pesquisas em plantas medicinais com atividade anti-Sporothrix spp. Um levantamento bibliográfico foi realizado com base em artigos científicos sobre a atividade antifúngica in vitro e in vivo de óleos essenciais e extratos de plantas preparados em diferentes solventes contra o complexo Sporothrix schenckii. A metodologia do estudo consistiu em uma revisão bibliográfica em Google Scholar, Science Direct, Pubmed, Bireme e Springer link com artigos desde 1986 até 2015. Foram encontradas 141 espécies de plantas já investigadas, das quais 100 espécies concentradas em 39 famílias botânicas apresentaram atividade anti-Sporothrix spp. confirmada. Combretaceae, Asteraceae e Lamiaceae representaram as famílias botânicas com maior número de espécies vegetais com potencial antifúngico, empregando diferentes metodologias. Entretanto, há poucos estudos com plantas medicinais em infecção experimental animal comprovando sua atividade no tratamento da esporotricose. Reforça-se a necessidade de mais pesquisas relacionadas à padronização de metodologias in vitro e a estudos in vivo relacionados à segurança e potencial tóxico dessas plantas com atividade anti-Sporothrix spp.

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Sporotrichosis is a zoonosis caused by fungi of the Sporothrix schenckii complex, whichSporothrix brasiliensis is prevalent in cats and dogs in Brazil. Itraconazole is the antifungal used for treatment,but the emergence of resistance has encouraged the search for therapeutic alternatives in medicinal plants.Among the benefits properties of green tea (Camellia sinensis L. Kuntze), it is included the antifungal activity,but there are no data that prove its potential in sporotrichosis. Aimed to evaluate the in vitro antifungal activityof green tea against Sporothrix brasiliensis. Infusion and decoction of dried leaves were prepared at 10% andtested through broth microdilution test (CLSI M38-A2), that was also performed with itraconazole. Nine clinicalisolates of S. brasiliensis from cats and dogs in South Brazil and one standard strain of S. schenckii were tested.The results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration(MFC). All isolates were inhibited by infusion and decoction (MIC90 of 3.125 mg mL-1for both extracts),without statistic difference, but the fungicidal activity was weak or no occurred (MFC90 > 50 mg mL-1for both).In itraconazole, S. brasiliensis were sensitive at MIC50 of 2 μg mL-1and S. schenckii to MIC of 0.0625 μg mL-1,but the antifungal resistance was observed (MIC90 and MFC50/90 > 16 μg mL-1). For the first time, the satisfactoryinhibitory activity of the infusion and decoction of green tea make its promising in the treatment ofsporotrichosis. However, more studies are needed for its use

Esporotricose é uma micose zoonótica causada por fungos do complexo Sporothrix schenckii, oqual Sporothrix brasiliensis é prevalente em gatos e cães no Brasil. Itraconazol é o antifúngico utilizado para otratamento, entretanto, o surgimento de resistência tem encorajado a busca por alternativas terapêuticas emplantas medicinais. Dentre as propriedades benéficas do chá-verde (Camellia sinensis L. Kuntze), inclue-seatividade antifúngica, embora não haja publicações que relacionem esse potencial na esporotricose. Esse estudoobjetivou avaliar a atividade antifúngica in vitro do chá-verde contra Sporothrix brasiliensis. Infusão e decocçãode folhas secas de chá-verde foram preparadas a 10% e testadas através do teste de microdiluição em caldo(CLSI M38-A2), também realizado com itraconazol. Nove isolados clínicos de S. brasiliensis oriundos de gatose cães com esporotricose na região sul do Rio Grande do Sul, Brasil, e uma cepa padrão de S. schenckii foramtestadas. Os resultados foram expressos em concentração inibitória mínima (CIM) e concentração fungicidamínima (CFM). Todos os isolados foram inibidos pela infusão e decocção (CIM90 de 3.125 mg mL-1para ambosextratos), sem diferença estatística, porém, a atividade fungicida foi fraca ou não ocorreu (CIM90 > 50 mg mL-1para ambos). Em itraconazol, S. brasiliensis foram sensíveis na CIM50 de 2 μg mL-1e S. schenckii à CIM de0.0625 μg mL-1, porém, a resistência antifúngica foi observada (CIM90 > 16 μg mL-1). Pela primeira vez, aatividade inibitória satisfatória da infusão e decocção de chá-verde os tornam promissores no tratamento daesporotricose. No entanto, mais estudos são necessários para seu uso

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Sporotrichosis is a zoonosis caused by fungi of the Sporothrix schenckii complex, whichSporothrix brasiliensis is prevalent in cats and dogs in Brazil. Itraconazole is the antifungal used for treatment,but the emergence of resistance has encouraged the search for therapeutic alternatives in medicinal plants.Among the benefits properties of green tea (Camellia sinensis L. Kuntze), it is included the antifungal activity,but there are no data that prove its potential in sporotrichosis. Aimed to evaluate the in vitro antifungal activityof green tea against Sporothrix brasiliensis. Infusion and decoction of dried leaves were prepared at 10% andtested through broth microdilution test (CLSI M38-A2), that was also performed with itraconazole. Nine clinicalisolates of S. brasiliensis from cats and dogs in South Brazil and one standard strain of S. schenckii were tested.The results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration(MFC). All isolates were inhibited by infusion and decoction (MIC90 of 3.125 mg mL-1for both extracts),without statistic difference, but the fungicidal activity was weak or no occurred (MFC90 > 50 mg mL-1for both).In itraconazole, S. brasiliensis were sensitive at MIC50 of 2 μg mL-1and S. schenckii to MIC of 0.0625 μg mL-1,but the antifungal resistance was observed (MIC90 and MFC50/90 > 16 μg mL-1). For the first time, the satisfactoryinhibitory activity of the infusion and decoction of green tea make its promising in the treatment ofsporotrichosis. However, more studies are needed for its use(AU)

Esporotricose é uma micose zoonótica causada por fungos do complexo Sporothrix schenckii, oqual Sporothrix brasiliensis é prevalente em gatos e cães no Brasil. Itraconazol é o antifúngico utilizado para otratamento, entretanto, o surgimento de resistência tem encorajado a busca por alternativas terapêuticas emplantas medicinais. Dentre as propriedades benéficas do chá-verde (Camellia sinensis L. Kuntze), inclue-seatividade antifúngica, embora não haja publicações que relacionem esse potencial na esporotricose. Esse estudoobjetivou avaliar a atividade antifúngica in vitro do chá-verde contra Sporothrix brasiliensis. Infusão e decocçãode folhas secas de chá-verde foram preparadas a 10% e testadas através do teste de microdiluição em caldo(CLSI M38-A2), também realizado com itraconazol. Nove isolados clínicos de S. brasiliensis oriundos de gatose cães com esporotricose na região sul do Rio Grande do Sul, Brasil, e uma cepa padrão de S. schenckii foramtestadas. Os resultados foram expressos em concentração inibitória mínima (CIM) e concentração fungicidamínima (CFM). Todos os isolados foram inibidos pela infusão e decocção (CIM90 de 3.125 mg mL-1para ambosextratos), sem diferença estatística, porém, a atividade fungicida foi fraca ou não ocorreu (CIM90 > 50 mg mL-1para ambos). Em itraconazol, S. brasiliensis foram sensíveis na CIM50 de 2 μg mL-1e S. schenckii à CIM de0.0625 μg mL-1, porém, a resistência antifúngica foi observada (CIM90 > 16 μg mL-1). Pela primeira vez, aatividade inibitória satisfatória da infusão e decocção de chá-verde os tornam promissores no tratamento daesporotricose. No entanto, mais estudos são necessários para seu uso(AU)

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Descrevem-se os aspectos clinicopatológicos de casos de aflatoxicose em cães no Sul do Rio Grande do Sul. Foi realizado um estudo retrospectivo dos casos diagnosticados como aflatoxicose em cães necropsiados no Laboratório Regional de Diagnóstico (LRD) da Faculdade de Veterinária da Universidade Federal de Pelotas (UFPel) no período de 1978 a 2012. Em quatro casos o diagnóstico foi confirmado pela detecção de níveis de 89 a 191 ppb de aflatoxinas B1 e G1 no alimento dos cães. De um total de 27 cães com cirrose hepática, em seis havia suspeita de aflatoxicose pelas lesões macro e microscópicas e pelo tipo de alimentação que os cães recebiam. Os sinais clínicos nos casos confirmados e nos suspeitos caracterizaram-se por apatia, diarreia, icterícia e ascite, com evolução para morte em 8 a 30 dias nos casos confirmados e em 15 a 60 dias nos casos suspeitos. A dieta era à base de derivados de milho ou arroz, farelo de amendoim e, em um caso suspeito, a dieta era ração comercial. As alterações macroscópicas caracterizaram-se por ascite, icterícia, fígado aumentado de tamanho, com ou sem nódulos, hemorragia nas serosas, conteúdo intestinal hemorrágico. Os casos foram classificados de acordo com o padrão histológico principal, caracterizado por vacuolização difusa no citoplasma de hepatócitos nos casos agudos, por proliferação de ductos biliares e discreta fibroplasia nos casos subagudos e por fibrose acentuada nos casos crônicos. Aparentemente, a enfermidade não é importante como causa de morte em cães na região, no entanto, alerta-se para a possibilidade de casos com diagnóstico de cirrose hepática sem causa determinada serem causados por aflatoxicose.

Clinical pathological aflatoxicosis in dogs is described in southern Rio Grande do Sul. It was conducted a retrospective study of cases diagnosed as aflatoxicosis in dogs necropsied at the Regional Diagnostic Laboratory (LRD) of the Veterinary School of the Federal University of Pelotas (UFPel) in the period 1978-2012. In four cases the diagnosis was confirmed by detection of levels of aflatoxins B1 and G1, with the finding of 89-191ppb in the feed. The macroscopic and histologic lesions and the diet observed in six of 27 dogs with liver cirrhosis led to suspicion of aflatoxicosis. Clinical signs evidenced in confirmed or suspected cases were lethargy, diarrhea, jaundice and ascites, progressing to death within 8 to 30 days in confirmed cases, and within 15 to 60 days in suspected cases. The diet was corn and rice byproducts and peanut meal, and one of the dogs received commercial ration. Gross changes were characterized by ascites, jaundice, enlarged liver, with or without regenerative nodules, hemorrhages in serous membranes and bloody intestinal content. The cases were classified according to the main histological pattern, characterized by diffuse vacuolation of the cytoplasm of hepatocytes in acute cases, by proliferation of bile ducts, and mild fibrosis in subacute cases, and by severe fibrosis in chronic cases. Apparently the disease is not important as a cause of death in dogs in the region, nevertheless the possibility of cases of cirrhosis of unknown etiology would be caused by aflatoxicosis.

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Descrevem-se os aspectos clinicopatológicos de casos de aflatoxicose em cães no Sul do Rio Grande do Sul. Foi realizado um estudo retrospectivo dos casos diagnosticados como aflatoxicose em cães necropsiados no Laboratório Regional de Diagnóstico (LRD) da Faculdade de Veterinária da Universidade Federal de Pelotas (UFPel) no período de 1978 a 2012. Em quatro casos o diagnóstico foi confirmado pela detecção de níveis de 89 a 191 ppb de aflatoxinas B1 e G1 no alimento dos cães. De um total de 27 cães com cirrose hepática, em seis havia suspeita de aflatoxicose pelas lesões macro e microscópicas e pelo tipo de alimentação que os cães recebiam. Os sinais clínicos nos casos confirmados e nos suspeitos caracterizaram-se por apatia, diarreia, icterícia e ascite, com evolução para morte em 8 a 30 dias nos casos confirmados e em 15 a 60 dias nos casos suspeitos. A dieta era à base de derivados de milho ou arroz, farelo de amendoim e, em um caso suspeito, a dieta era ração comercial. As alterações macroscópicas caracterizaram-se por ascite, icterícia, fígado aumentado de tamanho, com ou sem nódulos, hemorragia nas serosas, conteúdo intestinal hemorrágico. Os casos foram classificados de acordo com o padrão histológico principal, caracterizado por vacuolização difusa no citoplasma de hepatócitos nos casos agudos, por proliferação de ductos biliares e discreta fibroplasia nos casos subagudos e por fibrose acentuada nos casos crônicos. Aparentemente, a enfermidade não é importante como causa de morte em cães na região, no entanto, alerta-se para a possibilidade de casos com diagnóstico de cirrose hepática sem causa determinada serem causados por aflatoxicose.(AU)

Clinical pathological aflatoxicosis in dogs is described in southern Rio Grande do Sul. It was conducted a retrospective study of cases diagnosed as aflatoxicosis in dogs necropsied at the Regional Diagnostic Laboratory (LRD) of the Veterinary School of the Federal University of Pelotas (UFPel) in the period 1978-2012. In four cases the diagnosis was confirmed by detection of levels of aflatoxins B1 and G1, with the finding of 89-191ppb in the feed. The macroscopic and histologic lesions and the diet observed in six of 27 dogs with liver cirrhosis led to suspicion of aflatoxicosis. Clinical signs evidenced in confirmed or suspected cases were lethargy, diarrhea, jaundice and ascites, progressing to death within 8 to 30 days in confirmed cases, and within 15 to 60 days in suspected cases. The diet was corn and rice byproducts and peanut meal, and one of the dogs received commercial ration. Gross changes were characterized by ascites, jaundice, enlarged liver, with or without regenerative nodules, hemorrhages in serous membranes and bloody intestinal content. The cases were classified according to the main histological pattern, characterized by diffuse vacuolation of the cytoplasm of hepatocytes in acute cases, by proliferation of bile ducts, and mild fibrosis in subacute cases, and by severe fibrosis in chronic cases. Apparently the disease is not important as a cause of death in dogs in the region, nevertheless the possibility of cases of cirrhosis of unknown etiology would be caused by aflatoxicosis.(AU)

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Background: The dimorphic fungus Sporothrix schenckii and the yeasts of the Cryptococcus complex are important fungal pathogens of humans and animals that cause sporotrichosis and cryptococcosis, respectively. Both fungi are saprophytes of soils rich in organic matter and infect the host through traumatic injuries and/or inhalation. Sporotrichosis has been recognized as an important disease in the small animal clinic and in public health. Cases of this mycosis in cats and dogs have been reported in various Brazilian States. Our research group diagnosed eleven cases of sporotrichosis in dogs in Rio Grande do Sul in a five-year period. On the other hand, clinical cases of cryptococcosis in dogs and cats have not often been described in Brazil. In addition, infections caused by non- neoformans species have rarely been reported throughout the world. The aim of this study is to describe the first case of concomitant infection by Sporothrix schenckii and Cryptococcus albidus in a dog. Case: In 2008, a veterinary clinic of Rio Grande municipality received a 5-year-old female Dalmatian which had presented an increase of the nasal plan, sneezing, dyspnea and a serous nasal discharge for 30 days. Upon clinical examination, abnormalities in retal temperature, cardiopulmonary auscultation and frequency of the animal were not observed. The presence of an eritematous non-ulcerated mass partially occluding the airways was verified in the nasal cavity. Differential diagnosis included sporotrichosis, cryptococcosis and transmissible venereal tumor (TVT). Diagnosis confirmation was performed by laboratory tests which included radiographic, cytological and mycological analyses. The nasal radiograph revealed a mass of 20 mm in diameter with adjacent bone compromising. Ovoid and round cells suggestive of yeasts were found in cytology. Colonies with macroscopic and microscopic morphology compatible with Sporothrix schenckii and Cryptococcus albidus were obtained in mycological analysis. Confirmation of the species C. albidus was obtained by the API32 commercial system. Antifungal therapy with itraconazole at a 10 mg kg -1 oral dosage for 90 days was successfully performed, resulting in the regression of the lesions. Discussion: Sporotrichosis has been frequently diagnosed in cats and dogs in the south and southeast of Brazil. However, mixed infection cases with Sporothrix schenckii are rare. In Brazil, one case was described in a cat with sporotrichosis, demodicosis and pediculosis which was co-infected with the feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV). Mixed fungal infection by Cryptococcus neoformans and was described only in a dog in Canada that presented lesions similar to those reported in this study. Cryptococcosis cases in animals and humans caused by non-neoformans species with C. albidus and C. magnus have been described worldwide. Systemic, renal and cutaneous infections by C. albidus have been reported in dogs, cats and horses. The present study reports the first case of a concomitant infection of Sporothrix schenckii and Cryptococcus albidus in a dog, and warns small animal doctors of the importance of laboratory exams so that a final infectious disease diagnosis, especially of fungal infections, can be reached.

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