Granulomatous rhinitis by Neoconidiobolus lamprauges in a mule / Rinite granulomatosa por Neoconidiobolus lamprauges em uma mula

ABSTRACT: Conidiobolomycosis has a wide distribution, predominantly in humid tropical regions, affecting several species with significant mortality rates. The genus Conidiobolus is now divided into four genera: Capillidium, Conidiobolus, Microconidiobolus, and Neoconidiobolus. There are no confirmed reports of infection by these fungi in Equidae in Brazil. We present a rhinofacial rhinitis caused by Neoconidiobolus lamprauges in a mule from Rio de Janeiro, Brazil. The mule presented bilateral semi-occlusion of the nostrils, difficulty breathing, and weight loss. The histological examination of the nostril biopsied mass revealed multifocal necrotizing areas with nonstained images of fungal hyphae in the Splendore-Hoeppli reaction and surrounded by macrophages, eosinophils, neutrophils, and multinucleated giant cells. The Grocott methenamine silver staining revealed thin-walled, rarely septated, irregular branching hyphae, with a varying diameter of 12 μm (± 3.63 μm), and terminal ballooning dilations. The determining etiology of this rhinitis was based on the hyphae staining by immunohistochemistry and by amplifying the DNA fragment of N. lamprauges by polymerase chain reaction. Conidiobolomycosis should be included in the differential diagnosis of the causes of rhinitis in equids, mainly in tropical regions.

RESUMO: A conidiobolomicose apresenta ampla distribuição geográfica, com predominância em regiões tropicais úmidas, afeta várias espécies e apresenta taxa de mortalidade significativa. O gênero Conidiobolus está agora dividido em quatro gêneros: Capillidium, Conidiobolus, Microconidiobolus e Neoconidiobolus. Não há relatos confirmados de infecção por esses fungos em equídeos no Brasil. Relata-se o caso de uma mula com rinite rinofacial causada por Neoconidiobolus lamprauges no Rio de Janeiro, Brasil. A mula apresentava uma massa que semiocluía as narinas bilateralmente, dificuldade respiratória e emagrecimento. O exame histológico de biópsia da massa da narina revelou áreas de necrose multifocais com imagens negativas de hifas fúngicas em meio à reação de Splendore-Hoeppli, circundadas por macrófagos, eosinófilos, neutrófilos e células gigantes multinucleadas. O exame histoquímico metenamina prata de Grocott revelou hifas de parede fina, raramente septadas, com ramificação irregular, grau variável de paralelismo, e diâmetro médio de 12 μm (± 3,63) e dilatações balonosas terminais. O diagnóstico etiológico foi realizado pela associação da imuno-histoquímica e da amplificação do fragmento de DNA de N. lamprauges pela reação em cadeia da polimerase. A conidiobolomicose deve ser incluída no diagnóstico diferencial das causas de rinite em equídeos, principalmente em regiões tropicais.

Granulomatous rhinitis by Neoconidiobolus lamprauges in a mule / Rinite granulomatosa por Neoconidiobolus lamprauges em uma mula

Conidiobolomycosis has a wide distribution, predominantly in humid tropical regions, affecting several species with significant mortality rates. The genus Conidiobolus is now divided into four genera: Capillidium, Conidiobolus, Microconidiobolus, and Neoconidiobolus. There are no confirmed reports of infection by these fungi in Equidae in Brazil. We present a rhinofacial rhinitis caused by Neoconidiobolus lamprauges in a mule from Rio de Janeiro, Brazil. The mule presented bilateral semi-occlusion of the nostrils, difficulty breathing, and weight loss. The histological examination of the nostril biopsied mass revealed multifocal necrotizing areas with nonstained images of fungal hyphae in the Splendore-Hoeppli reaction and surrounded by macrophages, eosinophils, neutrophils, and multinucleated giant cells. The Grocott methenamine silver staining revealed thin-walled, rarely septated, irregular branching hyphae, with a varying diameter of 12 µm (± 3.63 µm), and terminal ballooning dilations. The determining etiology of this rhinitis was based on the hyphae staining by immunohistochemistry and by amplifying the DNA fragment of N. lamprauges by polymerase chain reaction. Conidiobolomycosis should be included in the differential diagnosis of the causes of rhinitis in equids, mainly in tropical regions.

A conidiobolomicose apresenta ampla distribuição geográfica, com predominância em regiões tropicais úmidas, afeta várias espécies e apresenta taxa de mortalidade significativa. O gênero Conidiobolus está agora dividido em quatro gêneros: Capillidium, Conidiobolus, Microconidiobolus e Neoconidiobolus. Não há relatos confirmados de infecção por esses fungos em equídeos no Brasil. Relata-se o caso de uma mula com rinite rinofacial causada por Neoconidiobolus lamprauges no Rio de Janeiro, Brasil. A mula apresentava uma massa que semiocluía as narinas bilateralmente, dificuldade respiratória e emagrecimento. O exame histológico de biópsia da massa da narina revelou áreas de necrose multifocais com imagens negativas de hifas fúngicas em meio à reação de Splendore-Hoeppli, circundadas por macrófagos, eosinófilos, neutrófilos e células gigantes multinucleadas. O exame histoquímico metenamina prata de Grocott revelou hifas de parede fina, raramente septadas, com ramificação irregular, grau variável de paralelismo, e diâmetro médio de 12 µm (± 3,63) e dilatações balonosas terminais. O diagnóstico etiológico foi realizado pela associação da imuno-histoquímica e da amplificação do fragmento de DNA de N. lamprauges pela reação em cadeia da polimerase. A conidiobolomicose deve ser incluída no diagnóstico diferencial das causas de rinite em equídeos, principalmente em regiões tropicais.

Aspergilose sistêmica em um cavalo - relato de caso / Systemic aspergilosis in a horse - case report

O presente trabalho relata o caso de aspergilose sistêmica em um cavalo da raça Quarto de Milha. O animal apresentava quadro de emagrecimento progressivo e alopecia. Os parâmetros avaliados durante o exame clínico encontravam-se de acordo com os valores normais para a espécie. No hemograma verificou-se leucocitose por neutrofilia, sem desvio à esquerda, proteínas plasmáticas totais elevadas e aumento do fibrinogênio, indicando um processo inflamatório acompanhado de desidratação. Os exames bioquímicos séricos demonstraram aumento no valor da ureia. Apesar dos exames realizados e da terapia instituída com antibióticos de amplo espectro e suporte nutricional, após 51 dias de internamento o quadro clínico evoluiu para caquexia e decúbito permanente do paciente, o qual foi submetido à eutanásia seguida de necropsia. Com base nos achados nos exames necroscópico e histopatológico, diagnosticou-se infecção sistêmica por Aspergillus sp., acometendo pulmões, fígado, rins, peritônio parietal e encéfalo. Dentre os fatores predisponentes à relatada infecção fúngica sistêmica, destaca-se o status imunológico debilitado do paciente, possivelmente proporcionado por tratamentos prolongados com antibióticos e anti-inflamatórios esteroidais. O presente trabalho relata o primeiro caso descrito de arpergilose sistêmica no Brasil. O diagnóstico definitivo só foi possível após necropsia e histopatologia, visto que os sinais clínicos inespecíficos não foram determinantes para o diagnóstico em vida.(AU)

This paper reports the case of systemic aspergillosis on a Quarter Horse. The animal had progressive weight loss and alopecia. The parameters evaluated during the clinical examination were in agreement with the normal values for the species. The hemogram showed neutrophilic leukocytosis, without left-sided, elevated total plasma proteins and an increase in fibrinogen, indicating an inflammatory process accompanied by dehydration. Serum biochemical tests showed an increase in the value of urea. Despite the performed tests and established therapy with broad spectrum antibiotics and nutritional support, after 51 days of hospitalization the clinical condition evolved into cachexia and permanent decubitus of the patient which underwent to euthanasia and necropsy. From the findings of necropsy and histopathology, systemic infection was diagnosed by Aspergillus sp., affecting the lungs, liver, kidneys, parietal peritoneum and encephalon. Among the factors predisposing to the reported systemic fungal infection, the patient's weakened immune status is highlighted, possibly due to prolonged antibiotic and steroidal anti-inflammatory treatments. This paper reports the first case described of systemic aspergillosis in Brazil. Definitive diagnosis was only possible after necropsy and histopathology, since nonspecific clinical signs were not determinant for the diagnosis in life.(AU)

Aspergilose sistêmica em um cavalo - relato de caso / Systemic aspergilosis in a horse - case report

O presente trabalho relata o caso de aspergilose sistêmica em um cavalo da raça Quarto de Milha. O animal apresentava quadro de emagrecimento progressivo e alopecia. Os parâmetros avaliados durante o exame clínico encontravam-se de acordo com os valores normais para a espécie. No hemograma verificou-se leucocitose por neutrofilia, sem desvio à esquerda, proteínas plasmáticas totais elevadas e aumento do fibrinogênio, indicando um processo inflamatório acompanhado de desidratação. Os exames bioquímicos séricos demonstraram aumento no valor da ureia. Apesar dos exames realizados e da terapia instituída com antibióticos de amplo espectro e suporte nutricional, após 51 dias de internamento o quadro clínico evoluiu para caquexia e decúbito permanente do paciente, o qual foi submetido à eutanásia seguida de necropsia. Com base nos achados nos exames necroscópico e histopatológico, diagnosticou-se infecção sistêmica por Aspergillus sp., acometendo pulmões, fígado, rins, peritônio parietal e encéfalo. Dentre os fatores predisponentes à relatada infecção fúngica sistêmica, destaca-se o status imunológico debilitado do paciente, possivelmente proporcionado por tratamentos prolongados com antibióticos e anti-inflamatórios esteroidais. O presente trabalho relata o primeiro caso descrito de arpergilose sistêmica no Brasil. O diagnóstico definitivo só foi possível após necropsia e histopatologia, visto que os sinais clínicos inespecíficos não foram determinantes para o diagnóstico em vida.(AU)

This paper reports the case of systemic aspergillosis on a Quarter Horse. The animal had progressive weight loss and alopecia. The parameters evaluated during the clinical examination were in agreement with the normal values for the species. The hemogram showed neutrophilic leukocytosis, without left-sided, elevated total plasma proteins and an increase in fibrinogen, indicating an inflammatory process accompanied by dehydration. Serum biochemical tests showed an increase in the value of urea. Despite the performed tests and established therapy with broad spectrum antibiotics and nutritional support, after 51 days of hospitalization the clinical condition evolved into cachexia and permanent decubitus of the patient which underwent to euthanasia and necropsy. From the findings of necropsy and histopathology, systemic infection was diagnosed by Aspergillus sp., affecting the lungs, liver, kidneys, parietal peritoneum and encephalon. Among the factors predisposing to the reported systemic fungal infection, the patient's weakened immune status is highlighted, possibly due to prolonged antibiotic and steroidal anti-inflammatory treatments. This paper reports the first case described of systemic aspergillosis in Brazil. Definitive diagnosis was only possible after necropsy and histopathology, since nonspecific clinical signs were not determinant for the diagnosis in life.(AU)

Infection of silkworm larvae by the entomopathogenic fungus Metarhizium anisopliae

ABSTRACT: The isolate E9 of Metarhizium anisopliae was used in commercial hybrids of Bombyx mori larvae to evaluate its biological effect. Symptomatological analyses showed typical signs of fungal infection. Histopathology revealed the presence of large numbers of hemocytes in the hemocoel, and on the sixth dpi the bodies of the insects appeared to be colonised by the fungus. The isolate E9 is pathogenic to larvae B. mori and; therefore, death of the insects was caused by the colonization of fungus in the epidermal and mesodermal tissues.

RESUMO: O isolado E9 de Metarhizium anisopliae foi usado em larvas híbridas de Bombyx mori para avaliar seu efeito biológico. A sintomatologia revelou sinais típicos de infecção por fungos. Na histopatologia foi verificado um aumento no número de hemócitos, sendo que, no 6 dpi, todo o corpo do inseto se apresentou colonizado pelo fungo. O isolado E9 é patogênico para lagartas de B. mori, causando sua morte pela colonização dos tecidos de origem epidérmica e mesodérmica.

Infection of silkworm larvae by the entomopathogenic fungus Metarhizium anisopliae / Infecção de lagartas do bicho-da-seda pelo fungo entomopatogênico Metarhizium anisopliae

The isolate E9 of Metarhizium anisopliae was used in commercial hybrids of Bombyx mori larvae to evaluate its biological effect. Symptomatological analyses showed typical signs of fungal infection. Histopathology revealed the presence of large numbers of hemocytes in the hemocoel, and on the sixth dpi the bodies of the insects appeared to be colonised by the fungus. The isolate E9 is pathogenic to larvae B. mori and; therefore, death of the insects was caused by the colonization of fungus in the epidermal and mesodermal tissues.

O isolado E9 de Metarhizium anisopliae foi usado em larvas híbridas de Bombyx mori para avaliar seu efeito biológico. A sintomatologia revelou sinais típicos de infecção por fungos. Na histopatologia foi verificado um aumento no número de hemócitos, sendo que, no 6 dpi, todo o corpo do inseto se apresentou colonizado pelo fungo. O isolado E9 é patogênico para lagartas de B. mori, causando sua morte pela colonização dos tecidos de origem epidérmica e mesodérmica.

Infection of silkworm larvae by the entomopathogenic fungus Metarhizium anisopliae / Infecção de lagartas do bicho-da-seda pelo fungo entomopatogênico Metarhizium anisopliae

The isolate E9 of Metarhizium anisopliae was used in commercial hybrids of Bombyx mori larvae to evaluate its biological effect. Symptomatological analyses showed typical signs of fungal infection. Histopathology revealed the presence of large numbers of hemocytes in the hemocoel, and on the sixth dpi the bodies of the insects appeared to be colonised by the fungus. The isolate E9 is pathogenic to larvae B. mori and; therefore, death of the insects was caused by the colonization of fungus in the epidermal and mesodermal tissues.(AU)

O isolado E9 de Metarhizium anisopliae foi usado em larvas híbridas de Bombyx mori para avaliar seu efeito biológico. A sintomatologia revelou sinais típicos de infecção por fungos. Na histopatologia foi verificado um aumento no número de hemócitos, sendo que, no 6 dpi, todo o corpo do inseto se apresentou colonizado pelo fungo. O isolado E9 é patogênico para lagartas de B. mori, causando sua morte pela colonização dos tecidos de origem epidérmica e mesodérmica.(AU)

Equine Sporotrichosis and iatrogenic hypothyroidism / Esporotricose equina e hipotireoidismo iatrogênico

Background: Sporotrichosis is a subacute or chronic granulomatous fungal infection that affects the dermis and subcutaneous tissue. Its etiology is related to a complex of Sporothrix spp. species of worldwide distribution. Usually, sporotrichosis results from traumatic inoculation of Sporothrix spp. into the dermis, affecting humans and other mammals, but a greater number of cases have been reported in felines, a fact increasingly important in the zoonotic transmission of diseased animals to man. In horses, there may be multiple ulcerated wounds covered with scabs and slightly elevated plaques or nodules that occur along lymphatic chains in limbs and can be unique and located. Iodides are considered drugs of choice for the treatment of sporotrichosis in horses. The main side effect of prolonged use of these substances is iodism and thyroid dysfunction. The aim of this study was to report a clinical case of sporotrichosis in an equine treated in the state of Paraná, southern Brazil, which showed secondary iatrogenic hypothyroidism with prolonged use of sodium and potassium iodide. Case: A female, 3-year-old, Quarter Horse, with 400 kg of liveweight presented at clinical examination of the cutaneous tissue elevated lesions of nodular aspect, with 2 to 4 cm in diameter, absence of itching and some nodules presenting alopecia areas in the chest and abdominal region, and in the [...](AU)

Equine Sporotrichosis and iatrogenic hypothyroidism / Esporotricose equina e hipotireoidismo iatrogênico

Background: Sporotrichosis is a subacute or chronic granulomatous fungal infection that affects the dermis and subcutaneous tissue. Its etiology is related to a complex of Sporothrix spp. species of worldwide distribution. Usually, sporotrichosis results from traumatic inoculation of Sporothrix spp. into the dermis, affecting humans and other mammals, but a greater number of cases have been reported in felines, a fact increasingly important in the zoonotic transmission of diseased animals to man. In horses, there may be multiple ulcerated wounds covered with scabs and slightly elevated plaques or nodules that occur along lymphatic chains in limbs and can be unique and located. Iodides are considered drugs of choice for the treatment of sporotrichosis in horses. The main side effect of prolonged use of these substances is iodism and thyroid dysfunction. The aim of this study was to report a clinical case of sporotrichosis in an equine treated in the state of Paraná, southern Brazil, which showed secondary iatrogenic hypothyroidism with prolonged use of sodium and potassium iodide. Case: A female, 3-year-old, Quarter Horse, with 400 kg of liveweight presented at clinical examination of the cutaneous tissue elevated lesions of nodular aspect, with 2 to 4 cm in diameter, absence of itching and some nodules presenting alopecia areas in the chest and abdominal region, and in the [...]

Epidemiological, clinical and diagnostic aspects of sheep conidiobolomycosis in Brazil / Aspectos epidemiológicos, clínicos e de diagnóstico da conidiobolomicose ovina no Brasil

Conidiobolomycosis is an emerging disease caused by fungi of the cosmopolitan genus Conidiobolus . Particular strains of Conidiobolus coronatus, Conidiobolus incongruus and Conidiobolus lamprauges , mainly from tropical or sub-tropical origin, cause the mycosis in humans and animals, domestic or wild. Lesions are usually granulomatous and necrotic in character, presenting two clinical forms: rhinofacial and nasopharyngeal. This review includes the main features of the disease in sheep, with an emphasis on the epidemiology, clinical aspects, and diagnosis of infections caused by Conidiobolus spp. in Brazil. In this country, the disease is endemic in the Northeast and Midwest, affecting predominantly woolless sheep breeds and occasioning death in the majority of the studied cases. The species responsible for infections of sheep are C. coronatus and C. lamprauges and the predominant clinical presentation is nasopharyngeal. These fungal infections are very important, since they compromise the health status of the sheep flock and cause serious economic losses to the sheep industry. Thus, research is needed to investigate faster tools for diagnosis and effective methods for the control and treatment of conidiobolomycosis.(AU)

Conidiobolomicose é uma enfermidade emergente causada por fungos cosmopolitas do gênero Conidiobolus . Isolados de Conidiobolus coronatus, Conidiobolus incongruus e Conidiobolus lamprauges, principalmente de origem tropical e subtropical, afetam humanos, animais domésticos e silvestres. As lesões da conidiobolomicose normalmente são de caráter granulomatoso e necrótico, apresentando-se sob duas formas clínicas: rinofacial e nasofaríngea. O presente artigo tem como objetivo revisar as principais características da doença em ovinos, particularizando a epidemiologia, assim como os aspectos clínicos e o diagnóstico das infecções causadas por Conidiobolus spp. no Brasil. Neste País, a enfermidade é endêmica nas regiões nordeste e centro-oeste, afetando ovinos predominantemente de raças deslanadas, ocasionando a morte na grande maioria dos casos estudados. As espécies do fungo responsáveis pelas infecções em ovinos são C. coronatus e C. lamprauges e a forma clínica predominante é a nasofaríngea. Tais infecções fúngicas são muito importantes, uma vez que comprometem o status sanitário do rebanho ovino e acarretam sérios prejuízos econômicos à ovinocultura. Dessa forma, pesquisas são necessárias para investigar ferramentas rápidas de diagnóstico e métodos eficazes de controle e tratamento da conidiobolomicose.(AU)

Caracterização de Malassezia spp isoladas de conduto auditivo de cães e gatos e avaliação dos fatores virulência

O gênero Malassezia compreende 17 espécies de leveduras lipofílicas e lipodependentes, componentes naturais da microbiota da pele dos animais homeotérmicos, incluindo humanos, e conhecido agente oportunista em dermatites e otites. A maior parte da casuística de otite fúngica em cães é atribuída a M. pachydermatis, mas espécies lipodependentes também podem ser agentes desta afecção, sendo possível que haja subdiagnósticos. Na rotina clínica veterinária, é comum o uso do exame direto de amostras otológicas como ferramenta auxiliar para direcionar uma melhor conduta terapêutica. Tendo esses pontos em vista, o trabalho objetiva realizar isolamento de Malassezia spp. a partir de amostras de otite de cães e gatos, identificando-as à nível de espécie, observar a presença da Malassezia em infeções mistas ou puras, verificar in vitro a produção dos principais fatores de virulência inerentes à patogenicidade e correlacionar a produção dos fatores de virulência com a natureza da infecção. Foram utilizadas 170 amostras oriundas dos condutos auditivos externos de 94 cães e 11 gatos com otite clínica, encaminhadas ao Laboratório de Leveduras Patogênicas e Ambientais, onde foram semeadas nos meios Agar Dixon Modificado e Agar Sabouraud Dextrose 2% com Cloranfenicol. Foram realizados testes clássicos para identificação fenotípica, como preconizado por Kurtzman et al. (2011) e análise proteômica por Maldi-TOF e bioquímico por VITEK2 ® para confirmação da identificação. Para análise da virulência foram testadas as produções de proteases, fosfolipase, lipases, esterases e hemolisina. Dos 105 animais, 63% (66) animais possuiam Malassezia sp. nas suas amostras, sendo uma lipodependente, e 37% (39) não tinham a levedura. O valor preditivo positivo e negativo do exame direto foram 75,58 e 67,19%, respectivamente. A sensibilidade do exame em relação ao isolamento foi de 75,58% e a especificidade de 67,19%. Não houve diferença estatística entre a recuperação das leveduras no diagnóstico em Agar Sabouraud Dextrose 2% e Agar Dixon Modificado. A identificação por espetrofotometria de massa teve 82% de acurácia na identificação dos isolados e as restantes não tiveram scores de identificação, enquanto o VITEK ® , identificou erroneamente todas as amostras. Para os testes enzimáticos, 100% (96) das amostras foram positivas para a produção de proteases e lipases. Apenas uma amostra não foi produtora de fosfolipase e esterase. 89% (86) amostras foram positivas para a produção de hemolisina. Conclui-se que a utilização do Agar Dixon Modificado é importante na rotina laboratorial de diagnóstico microbiologico de otites canina e felina, para ampliar a gama de espécies de Malassezia diagnosticadas. O exame direto mostrou ser um bom indicador para a presença da levedura, mas não necessáriamente para indicar seu envolvimento na infecção. Para além, percebe-se que essas leveduras in vitro possuem alta atividade enzimática, podendo fazer parte da patogênese da otite microbiana, mesmo quando não é o principal agente envolvido, como nas otites mistas, mesmo as que possuem contagens baixas no isolamento.

The genus Malassezia currently comprises 17 species of lipophilic and lipid- dependent yeasts. Considered a normal part of the skin microbiota of warm-blooded vertebrates, including humans, these may also become opportunistic agents in dermatitis and otitis. Fungal otitis in dogs have been associated with M. pachydermatis, but lipid- dependent species are also commonly found, indicating a possibility of frequent misdiagnosis. In the veterinary clinical routine, microscopy of otological samples is used as an auxiliary tool towards the implementation of an adequate drug therapy. The aim of this work was the isolation of Malassezia spp. from otitis samples from dogs and cats, their respective identification at the species level, observe the presence of Malassezia in pure or co-infections, in vitro detection the main virulence factors inherent to the genus pathogenicity being produced, and the establishment of a correlation between the detected virulence factors being produced and the nature of the infection. A total of 170 samples from the external ear canal of 94 dogs and 11 cats with clinical otitis were sent to the Laboratory of Pathogenic and Environmental Yeasts, where they were inoculated in Modified Dixon Agar and Sabouraud Dextrose 2% Agar supplemented with chloramphenicol. Classical methods allowed for the phenotypic identification, as recommended by Kurtzman et al. (2011) further corroborated through proteomic analysis by MALDI-TOF MS and biochemical analysis by VITEK2 ® . For virulence analysis, protease, phospholipase, lipase, esterase and hemolysin production were tested. Of the 105 animals, 63% (66) individuals were positive for Malassezia sp., as one lipid dependent, and 37% (39) were negative for the genus. The positive and negative predictive values of the direct exam were 75.58% and 67.19%, respectively. The sensitivity of the isolation test was 75.58% and the specificity 67.19%. There was no significant difference between yeast recovery in Sabouraud Dextrose 2% Agar and Modified Dixon Agar. The identification by mass spectrophotometry had an 82% accuracy in the identification of the isolates and remaining isolates did not reach the identification score, whereas VITEK® misidentify all the samples. For the enzymatic tests, 100% (96) of the samples were positive for the production of proteases and lipases. Only one sample did not produce phospholipase nor esterase. 89% (86) samples were hemolysin producers. It was possible to conclude that the use of Modified Dixon Agar is important in the laboratory routine for microbiological diagnosis of canine and feline otitis, extending the range of Malassezia species diagnosed. Direct examination proved to be a good indicator of the presence of yeast, but not enlightening regarding its involvement in the infection. In addition, these yeasts have high enzymatic activity in vitro and may be part of the pathogenesis of microbial otitis, even when it is not the main pathological agent, as in co-infections otitis even with low isolation counts. Key words: malasseziosis, microbiological diagnosis, fungal infection

Criptococose cutânea associada à severa infestação por Sarcoptes sp. e Demodex sp. em um cão / Cutaneous cryptococcosis associated with severe infestation by Sarcoptes sp. and Demodex sp. in a dog

Este trabalho descreve um caso de criptococose canina localizada na pele da região interdigital de um cão. Um cão de rua, sem raça definida, adulto jovem apresentava lesões pruriginosas, alopécicas e crostosas em toda extensão corpórea, e uma lesão ulcerada interdigital no membro pélvico esquerdo. A citologia aspirativa por agulha fina (CAAF) da lesão interdigital demonstrou um processo inflamatório acrescido de organismos leveduriformes compatíveis com Cryptococcus sp. Efetuou-se, a seguir, a cultura fúngica, que ratificou o resultado da CAAF. O paciente também apresentou diagnóstico positivo para Sarcoptes sp. e Demodex sp. Cryptococcus sp. é o agente etiológico da criptococose, uma doença infecciosa oportunista que acomete o homem, animais domésticos e silvestres. A provável imunidade baixa do cão, neste estudo, pode ter sido um fator desencadeante para o desenvolvimento da doença. A criptococose em caninos, apresentada na forma cutânea, é incomum, o que justifica a relevância deste relato.

The aim of this study was to report a case of cutaneous cryptococcosis located in the interdigital skin of a dog. A young adult mongrel dog presented pruritic alopecic crusty lesions throughout corporeal extension and an ulcerated lesion in the interdigital region of the left pelvic limb. A fine needle aspiration cytology (FNAC) of the interdigital lesion showed an inflammatory process plus yeast organisms similar to Cryptococcus sp. A mycological culture was carried out confirming the results of FNAC. The patient also had positive diagnosis to Sarcoptes sp. and Demodex sp.. Cryptococcus sp. is the etiologic agent of cryptococcosis, an infectious opportunistic disease that affects humans, domestic and wild animals. The probable low immunity of the dog, in this study, may have been a triggering factor for the development of the disease. Canine cutaneous cryptococcosis is unusual, which explains the relevance of this report case.

Criptococose cutânea associada à severa infestação por Sarcoptes sp. e Demodex sp. em um cão / Cutaneous cryptococcosis associated with severe infestation by Sarcoptes sp. and Demodex sp. in a dog

Este trabalho descreve um caso de criptococose canina localizada na pele da região interdigital de um cão. Um cão de rua, sem raça definida, adulto jovem apresentava lesões pruriginosas, alopécicas e crostosas em toda extensão corpórea, e uma lesão ulcerada interdigital no membro pélvico esquerdo. A citologia aspirativa por agulha fina (CAAF) da lesão interdigital demonstrou um processo inflamatório acrescido de organismos leveduriformes compatíveis com Cryptococcus sp. Efetuou-se, a seguir, a cultura fúngica, que ratificou o resultado da CAAF. O paciente também apresentou diagnóstico positivo para Sarcoptes sp. e Demodex sp. Cryptococcus sp. é o agente etiológico da criptococose, uma doença infecciosa oportunista que acomete o homem, animais domésticos e silvestres. A provável imunidade baixa do cão, neste estudo, pode ter sido um fator desencadeante para o desenvolvimento da doença. A criptococose em caninos, apresentada na forma cutânea, é incomum, o que justifica a relevância deste relato.(AU)

The aim of this study was to report a case of cutaneous cryptococcosis located in the interdigital skin of a dog. A young adult mongrel dog presented pruritic alopecic crusty lesions throughout corporeal extension and an ulcerated lesion in the interdigital region of the left pelvic limb. A fine needle aspiration cytology (FNAC) of the interdigital lesion showed an inflammatory process plus yeast organisms similar to Cryptococcus sp. A mycological culture was carried out confirming the results of FNAC. The patient also had positive diagnosis to Sarcoptes sp. and Demodex sp.. Cryptococcus sp. is the etiologic agent of cryptococcosis, an infectious opportunistic disease that affects humans, domestic and wild animals. The probable low immunity of the dog, in this study, may have been a triggering factor for the development of the disease. Canine cutaneous cryptococcosis is unusual, which explains the relevance of this report case.(AU)

Feo-hifomicose sistêmica em cão

The objective of this study was to describe clinical, laboratorial and histopathological findings of systemic phaeohyphomycosis in a dog. The animal was presented with a history of fatigue, ascites, lethargy, weight and appetite loss and polydipsia. The therapy for erliquiose and babesiosis was started. After 11 days, the dog returned with severe liver disease and died after forty-eight hours. At necropsy, there were black nodules in various abdominal organs. Histologically, necrotizing granulomatous reaction with hemorrhage associated with myriads of intralesional pigmented fungal hyphae was observed in multiple organs, characterizing systemic phaeohyphomycosis. Thus, it is important to include this illness in the differential diagnosis of severe liver diseases in dogs with ascites.

Descrevem-se os achados clínicos, laboratoriais e histopatológicos de feo-hifomicose sistêmica em um cão. Clinicamente observou-se cansaço, ascite, apatia, hiporexia, perda de peso e polidipsia. O animal estava sendo tratado para erliquiose e babesiose. Após 11 dias, o cão retornou com hepatopatia grave e veio a óbito em 48 horas. Na necropsia, observaram-se nódulos enegrecidos em vários órgãos da cavidade abdominal. Histologicamente, havia reação granulomatosa necrosante com hemorragia, associada a miríades de hifas fúngicas pigmentadas intralesionais em múltiplos órgãos, caracterizando feo-hifomicose sistêmica. Ressalta-se a importância de incluir esta doença no diagnóstico diferencial de hepatopatias graves em cães com ascite.

Feo-hifomicose sistêmica em cão

The objective of this study was to describe clinical, laboratorial and histopathological findings of systemic phaeohyphomycosis in a dog. The animal was presented with a history of fatigue, ascites, lethargy, weight and appetite loss and polydipsia. The therapy for erliquiose and babesiosis was started. After 11 days, the dog returned with severe liver disease and died after forty-eight hours. At necropsy, there were black nodules in various abdominal organs. Histologically, necrotizing granulomatous reaction with hemorrhage associated with myriads of intralesional pigmented fungal hyphae was observed in multiple organs, characterizing systemic phaeohyphomycosis. Thus, it is important to include this illness in the differential diagnosis of severe liver diseases in dogs with ascites.

Descrevem-se os achados clínicos, laboratoriais e histopatológicos de feo-hifomicose sistêmica em um cão. Clinicamente observou-se cansaço, ascite, apatia, hiporexia, perda de peso e polidipsia. O animal estava sendo tratado para erliquiose e babesiose. Após 11 dias, o cão retornou com hepatopatia grave e veio a óbito em 48 horas. Na necropsia, observaram-se nódulos enegrecidos em vários órgãos da cavidade abdominal. Histologicamente, havia reação granulomatosa necrosante com hemorragia, associada a miríades de hifas fúngicas pigmentadas intralesionais em múltiplos órgãos, caracterizando feo-hifomicose sistêmica. Ressalta-se a importância de incluir esta doença no diagnóstico diferencial de hepatopatias graves em cães com ascite.

Infecção fúngica mista por Sporothrix schenckii e Cryptococcus albidus em um canino / Mixed fungal infection by Sporothrix schenckii and Cryptococcus albidus in a dog

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.(AU)

Infecção fúngica mista por Sporothrix schenckii e Cryptococcus albidus em um canino / Mixed fungal infection by Sporothrix schenckii and Cryptococcus albidus in a dog

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.

Alopecia por Malassezia pachydermatis em um lobo-marinhosubantártico(Arctocephalus tropicalis) - relato de caso / Alopecia due to Malassezia pachydermatis in a sub-antartic fur seal(Arctocephalus tropicalis) - case report / Alopecia por Malassezia pachydermatis en un lobo marino subantártico(Arctocephalus tropicalis) - relato de caso

Mamíferos marinhos em cativeiro são particularmente suscetíveis a infecções virais, bacterianas, fúngicas ou parasitárias que causam dermatite focal ou difusa, e que podem causar alopecias em pinípedes. Diversos tipos de lesões dermatológicas ocasionadas por fungos, incluindo lesão por Malassezia, já foram descritas nesses animais. Um lobo-marinho-subantártico (Arctocephalus tropicalis) mantido em cativeiro apresentou alopecia bilateral na região da nadadeira caudal e em região abaixo da nadadeira peitoral. Para diagnosticar a possível causa da alopecia foram realizados etograma (para avaliação do comportamento do animal), culturas bacteriana e micológica. Após confirmação da presença de Malassezia pachydermatis, foi estabelecido protocolo de tratamento, o qual se mostrou bastante eficaz para a resolução do quadro. O objetivo do presente trabalho é relatar um caso de alopecia em um lobo-marinho, abordando o diagnóstico e a terapêutica utilizados.

Marine mammals in captivity are particularly susceptible to viral, bacterial, fungal or parasitic infections that can cause focal or diffuse dermatitis, and may cause alopecia in pinnipeds. Various types of skin lesions caused by fungi have been described in these animals, including lesions due to Malassezia. A subantartic fur seal (Arctocephalus tropicalis) kept in captivity presented bilateral alopecia on the caudal flipper and on the region below the pectoral flipper. To diagnose the possible cause of alopecia, bacterial and mycological cultures were performed, as well as an ethogram to assess the behavior of the animal. After confirmation of the presence of Malassezia pachydermatis in the affected regions, a treatment protocol was established, which proved to be effective for the resolution of the clinical case. The objective of this study is to report a case of alopecia in a fur seal, addressing the diagnosis and treatment that were performed.

Los mamíferos marinos en cautiverio son particularmente susceptibles a infecciones virales, bacterianas, fúngicas o parasitarias que provocan una dermatitis focal o difusa, y pueden causar alopecia en los pinnípedos. Varios tipos de lesiones de piel causadas por hongos, incluyendo lesiones por Malassezia, han sido descriptas en estos animales. Un lobo marino subantártico (Arctocephalus tropicalis) mantenido en cautiverio presentó una alopecia bilateral en la región de la aleta caudal y de la región por debajo de la aleta pectoral. Para diagnosticar la posible causa de la alopecia se realizó un etograma (para evaluar el comportamiento del animal), y cultivos bacterianos y micológicos. Después de confirmada la presencia de Malassezia pachydermatis, se estableció un protocolo de tratamiento que resultó muy eficaz para la resolución del cuadro. El objetivo de este trabajo es presentar un caso de alopecia en un lobo marino, abordando el diagnóstico y la conducta terapéutica utilizada.

Alopecia por Malassezia pachydermatis em um lobo-marinhosubantártico(Arctocephalus tropicalis) - relato de caso / Alopecia due to Malassezia pachydermatis in a sub-antartic fur seal(Arctocephalus tropicalis) - case report / Alopecia por Malassezia pachydermatis en un lobo marino subantártico(Arctocephalus tropicalis) - relato de caso

Mamíferos marinhos em cativeiro são particularmente suscetíveis a infecções virais, bacterianas, fúngicas ou parasitárias que causam dermatite focal ou difusa, e que podem causar alopecias em pinípedes. Diversos tipos de lesões dermatológicas ocasionadas por fungos, incluindo lesão por Malassezia, já foram descritas nesses animais. Um lobo-marinho-subantártico (Arctocephalus tropicalis) mantido em cativeiro apresentou alopecia bilateral na região da nadadeira caudal e em região abaixo da nadadeira peitoral. Para diagnosticar a possível causa da alopecia foram realizados etograma (para avaliação do comportamento do animal), culturas bacteriana e micológica. Após confirmação da presença de Malassezia pachydermatis, foi estabelecido protocolo de tratamento, o qual se mostrou bastante eficaz para a resolução do quadro. O objetivo do presente trabalho é relatar um caso de alopecia em um lobo-marinho, abordando o diagnóstico e a terapêutica utilizados.(AU)

Marine mammals in captivity are particularly susceptible to viral, bacterial, fungal or parasitic infections that can cause focal or diffuse dermatitis, and may cause alopecia in pinnipeds. Various types of skin lesions caused by fungi have been described in these animals, including lesions due to Malassezia. A subantartic fur seal (Arctocephalus tropicalis) kept in captivity presented bilateral alopecia on the caudal flipper and on the region below the pectoral flipper. To diagnose the possible cause of alopecia, bacterial and mycological cultures were performed, as well as an ethogram to assess the behavior of the animal. After confirmation of the presence of Malassezia pachydermatis in the affected regions, a treatment protocol was established, which proved to be effective for the resolution of the clinical case. The objective of this study is to report a case of alopecia in a fur seal, addressing the diagnosis and treatment that were performed.(AU)

Los mamíferos marinos en cautiverio son particularmente susceptibles a infecciones virales, bacterianas, fúngicas o parasitarias que provocan una dermatitis focal o difusa, y pueden causar alopecia en los pinnípedos. Varios tipos de lesiones de piel causadas por hongos, incluyendo lesiones por Malassezia, han sido descriptas en estos animales. Un lobo marino subantártico (Arctocephalus tropicalis) mantenido en cautiverio presentó una alopecia bilateral en la región de la aleta caudal y de la región por debajo de la aleta pectoral. Para diagnosticar la posible causa de la alopecia se realizó un etograma (para evaluar el comportamiento del animal), y cultivos bacterianos y micológicos. Después de confirmada la presencia de Malassezia pachydermatis, se estableció un protocolo de tratamiento que resultó muy eficaz para la resolución del cuadro. El objetivo de este trabajo es presentar un caso de alopecia en un lobo marino, abordando el diagnóstico y la conducta terapéutica utilizada.(AU)

Similarity between the in vitro activity and toxicity of two different fungizone™ / lipofundin™ admixtures

PURPOSE: Amphotericin B (AmB), an antifungal agent that presents a broad spectrum of activity, remains the gold standard in the antifungal therapy. However, sometimes the high level of toxicity forbids its clinical use. The aim of this work was to evaluate and compare the efficacy and toxicity in vitro of Fungizon (AmB-D) and two new different AmB formulations. METHODS: three products were studied: Fungizon, and two Fungizon /Lipofundin admixtures, which were diluted through two methods: in the first one, Fungizon was previously diluted with water for injection and then, in Lipofundin (AmB-DAL); the second method consisted of a primary dilution of AmB-D as a powder in the referred emulsion (AmB-DL). For the in vitro assay, two cell models were used: Red Blood Cells (RBC) from human donors and Candida tropicallis (Ct). The in vitro evaluation (K+ leakage, hemoglobin leakage and cell survival rate-CSR) was performed at four AmB concentrations (from 50 to 0.05mg.L-1). RESULTS: The results showed that the action of AmB was not only concentration dependent, but also cellular type and vehicle kind dependent. At AmB concentrations of 50 mg.L-1, although the hemoglobin leakage for AmB-D was almost complete (99.51), for AmB-DAL and AmB-DL this value tended to zero. The p = 0.000 showed that AmB-D was significantly more hemolytic. CONCLUSION: The Fungizon-Lipofundin admixtures seem to be the more valuable AmB carrier systems due to their best therapeutic index presented.

OBJETIVO: A anfotericina B é um agente antifúngico de largo espectro bastante empregado na terapia antifúngica. Entretanto, esta molécula apresenta um alto nível de toxicidade que, na maioria das vezes, impede o seu uso contínuo na terapêutica médica. O objetivo deste artigo foi comparar a eficácia e a toxicidade in vitro do Fungizon (AmB-D) e de dois sistemas carreadores de AmB. MÉTODOS: Três produtos foram avaliados: o Fungizon , e dois sistemas oriundos da mistura entre o Fungizon e o Lipofundin , uma emulsão de uso parenteral. Tais sistemas foram obtidos por duas técnicas: Na primeira diluiu-se previamanete o Fungizon com água para injetáveis e em seguida inseriu-se o Lipofundin (AmB-DAL); o segundo método consistiu na diluíção extemporânea do Fungizon com a referida emulsão (AmB-DL). Dois modelos celulares foram empregados no estudo: os eritrócitos (RBC) oriundos de doadores humanos e a Candida tropicalis (Ct). A avaliação in vitro (liberação de K+ e hemoglobina, e o índice de sobrevivência celular-CSR) foi realizado com quatro concentrações de AmB (entre 50 e 0.05mg.L-1). RESULTADOS: Os resultados demonstram que a ação da AmB não só foi dependente da concentração como também variou de acordo com o modelo celular e o veículo que diluiu o Fungizon . Nas concentrações de 50 mg.L-1, apesar da liberação de hemoglobina ser quase que total para AmB-D (99.51), para a AmB-DAL e AmB-DL este valor tendeu a zero. Um p = 0.000 demonstrou que AmB-D foi significativamente mais hemolítico. CONCLUSÃO: A mistura Fungizon -Lipofundin aparenta ser um bom sistema para carrear a AmB tendo em vista seu elevado índice terapêutico demonstrado.