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Sporotrichosis is a disease not requiring jurisdictional notification and consequently is underreported in Brazil. Therefore, the epidemiological picture even in hyperendemic states is unknown. Thus, we evaluated the occurrence of sporotrichosis throughout the territory of the southern state of Brazil, Rio Grande do Sul (RS).We update the epidemiological situation of sporotrichosis in the southern region of this state and describe the emergence of this disease in the Metropolitan region. We engaged professionals from RS enrolled in animal health care in answering a questionnaire regarding sporotrichosis.The occurrence of local cases of feline sporotrichosis was reported by 83% of the participants from 40 cities, distributed through the seven health districts of RS. Human sporotrichosis cases, transmitted by cats, were also reported by professionals from four regions of the state. The frequency of the disease in both the South and Metropolitan regions showed a marked increase in recent years.Feline and cat-transmitted human sporotrichosis is an underreported mycosis in RS, widely distributed in the territory of this state and increasing. Aggressive public health policies are urgently necessary to control the geographical expansion of this spreading mycosis.

Despite thousands of sporotrichosis cases related to the zoonotic transmission of Sporothrix brasiliensis have been described, the diagnostic gold standard is still the classical culturing methods. The mycological culture results are available afterseven to 30 days of incubation. Since an early diagnosis contributes to improving thetreatment and to the spread control of this mycosis, studies evaluating faster diagnosticmethods are needed. Therefore, we aimed to evaluate the species-specific PCR for thesporotrichosis diagnosis caused by S. brasiliensis, using samples collected with a noninvasive technique. Seventy-four swabs from feline (n=64), canine (n=5), and human(n=5) suspect sporotrichosis cases were included. All samples were submitted to classicalmethods for diagnosis and to S. brasiliensis-specific PCR. Using mycological culture asthe gold standard, the diagnosis of S. brasiliensis-caused infection was confirmed in 69%(51/74) of the cases. PCR was positive in 30 out of these 51 cases, showing 59%sensitivity, 100% specificity, 72% accuracy, 100% positive predictive value, and 52%negative predictive value. Consequently, more studies are needed to elucidate theinterference factors that culminated with the high rate of false-negative results and thenoptimize this molecular test for the accurate diagnosis of infections caused by S.brasiliensis.

We aimed to evaluate the chemical, antioxidant, cytotoxic, and antifungal activities of hydroalcoholic extracts of native plants from Southern Brazil: Schinus terebinthifolia (SCH), Persicaria hydropiperoides (PER), Eugenia uniflora (EUG) and Equisetum hyemale (EQU). Ethyl gallate, quercetin, and quinic acid were prevalent compounds identified by LC-MS. For total phenolic/flavonoid contents and the antioxidant potential against ABTS/DPPH radicals, the ascending order was EQU 75%) at concentrations up to 12.5 mg/mL (MTT assay). By M38-A2/M27-A3 (CLSI) against 68 clinical isolates of animals and strains of Malassezia pachydermatis, Sporothrix brasiliensis, Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes, all extracts (MIC/MFC ≤3.13–100 mg/mL) were active, except EUG. SCH inhibited and killed S. brasiliensis (MIC/MFC50/90 3.12–12.5 mg/mL) and dermatophytes (MIC/MFC 6.25–25 mg/mL) resistant to ketoconazole and itraconazole. These findings support the promising use of the selected plant extracts as antifungal agents.

Feline sporotrichosis is a relevant mycose in veterinary medicine due to its severity and zoonotic potential and the fact that it can be difficult to treat. The immune status of the animal exerts influence on the prognosis of the disease and determines its clinical outcome. This study evaluated the efficacy of the immunomodulatory thymomodulin as an adjunct to antifungal therapy in cats with disseminated sporotrichosis; thymomodulin was used in association with itraconazole (ITL) and potassium iodide (KI) to treat this fungal disease in the feline patient. Thirty-one cats (n=31) diagnosed with disseminated cutaneous sporotrichosis were divided into two groups as follows: Group 1 (G1) (n=16), which included those animals that were treated with thymomodulin in association with ITL and KI, and Group 2 (G2) (n=15) which had pacientsthat received ITL and KI only. The response to different treatment modalities was assessed, considering the survival rate, time frame for the lesions to respond to therapy, and clinical improvement or deterioration according to a body condition score system. Animals from G1 had a survival rate of nearly 100% (93.6%) that was approximately twice higher than the survival rate of those animals from G2 (53%). Moreover, patients from G1 had a significantly better prognosis, improved body condition, and shorter time for remission of the extra cutaneous clinical signs (p

Itraconazole is the first drug of choice for the treatment of sporotrichosis and it is available at different concentrations for veterinary patients. However, therapeutic failure has been reported, limiting clinical treatment. This study evaluated the in vitro efficacy of brand-name and compounded itraconazole formulations against Sporothrix brasiliensis and estimated the itraconazole content in each tested formulation. Oral capsules were acquired from two brand-name products for human (H-IND) and veterinary (V-IND) uses, and three from compounding pharmacies in Pelotas, RS, for human (H-COMP1/H-COMP2) and veterinary (V-COMP) uses. Capsule purity was analyzed by liquid chromatography–electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS). Antifungal activity was determined against 29 Sporothrix brasiliensis by the M38-A2 guideline of CLSI. H-IND/H-COMP1/H-COMP2 had high efficacy against S. brasiliensis (approximately 70% of total isolated susceptible), V-COMP showed moderate efficacy (51.7%), and V-IND was the least effective formulation (37.9%). Thirty-four percent of the total isolates were resistant to all formulations. Furthermore, itraconazole content did not match the concentration indicated by the manufacturers, ranging from 387.70 to 7.81 μg/mg (H-COMP2 > V-COMP > H-IND > H-COMP1 > V-IND). Therefore, it is possible that the formulations showed different in vitro efficacy due to the difference in their itraconazole contents. Given the emergence of antifungal resistance for all formulations, the choice product to be used must follow susceptibility testing. Stringent quality control measures are recommended for product manufactures to assure drug content uniformity.

Oregano (Origanum vulgare) has anti-Sporothrix spp. activity, including against strains that are resistant to antifungal drugs. As the studies are limited to the essential oil, the aim of this study was to evaluate the chemical, antioxidant and cytotoxic properties of polar oregano extracts and their anti-Sporothrix brasiliensis activity. Aerial plant parts were used in the preparation of 10 min (INF10) and 60 min (INF60) infusions, a decoction (DEC) and a hydroalcoholic extract (HAE). Six phenolic acids and four flavonoids were identified and quantified through liquid-chromatography (LC-MS). Extracts in increasing order of total phenolic and flavonoid contents were HAEINF60INF10DEC and HAEDECINF60INF10, respectively. All extracts showed DPPH and ABTS radical scavenging potential (HAEINF60DECINF10). HAE showed the least toxicity toward MDCK and CRFK cell lines in the MTT colorimetric assay. The antifungal potential against 29 S. brasiliensis isolates obtained from cats and dogs was evaluated following the CLSI M38-A2 protocol adapted to natural products. Minimum inhibitory concentration for 50% of the isolates (MIC50) was 5 mg/ml for all extracts, and minimum fungicidal concentration for 50% of the isolates (MFC50) was 20 mg/ml for INF10 and 40 mg/ml for the remaining extracts. MIC90 was 10 mg/ml for all extracts, except for DEC (20 mg/ml). MFC90 was 40 mg/ml for INF10 and 40 mg/ml for the other extracts. This is the first report of oregano extracts showing antifungal activity against S. brasiliensis. Its use in the treatment of sporotrichosis may be considered upon toxicity and safe-use conditions are tested.

Resumo Este estudo avaliou a ação in vitro e in vivo de extratos de Eugenia uniflora em nematódeos gastrintestinais de ovinos. No teste de inibição da eclodibilidade dos ovos, extratos aquosos e hidroalcoólicos foram testados nas concentrações de 40 a 1,25 mg/mL. O efeito citotóxico foi mensurado através do ensaio de MTT em células VERO nas concentrações de 4000 a 1,95 µg/mL. A composição química foi analisada através da marcha fitoquímica qualitativa. No teste in vivo, foram administrados 100 mg/kg, por via oral, do extrato hidroalcoólico por três dias em ovinos naturalmente infectados e realizada coleta de fezes para estimar a redução de ovos por grama de fezes (OPG). Diferentes extratos de E. uniflora inibiram a eclodibilidade dos ovos, com percentual de inibição variando de 14,56 a 99,75%, sendo os hidroalcoólicos mais promissores que os aquosos. Na marcha fitoquímica, compostos com ação anti-helmíntica comprovada foram observados, como flavonoides, saponinas, taninos e triterpenos. Nas condições testadas, os extratos apresentaram citotoxicidade nula a moderada. In vivo, a redução do OPG foi observada apenas no grupo controle (anti-helmíntico). Os extratos de E. uniflora mostraram-se promissores com ação in vitro, necessitando estudos que avaliem outras concentrações e formas de administração in vivo. Palavras-chave: planta medicinal; Haemonchus contortus; ruminantes; anti-helmíntico; pitangueira.

Zoonotic sporotrichosis has undergone a geographical expansion in Brazil in the last decade. Rio Grande do Sul (RS) is the second state of the country in which a number of feline sporotrichosis cases have been described. Since cats are the main zoonotic source of infection, this study aimed to describe 100 cases of human sporotrichosis occurring in the last 5 years in the southern region of RS, Brazil. In addition, we aimed to illustrate the zoonotic importance of the disease, describing four cases in the same family due to transmission by their cat. This great number of human cases in a short period of evaluation highlights the severity of sporotrichosis as a public health problem in the region, suggesting that a possible outbreak is occurring that requires immediate public intervention actions to weaken its impact.

A esporotricose e uma micose, subcutânea e zoonotica, causada pelo fungo dimorfico Sporothrix schenckii que afeta animais e humanos. A micose em caes era considerada rara devido aos poucos relatos descritos na literatura, no entanto, nos ultimos anos observou-se um crescente registro de casos desta doenca no Brasil. Neste estudo foram analisadas as caracteristicas clinicas da esporotricose em tres caes diagnosticados no Laboratorio de Micologia (FV-UFPel), os quais apresentavam dispneia, secrecao nasal e espirros. Dois destes demonstravam tambem, lesoes cutâneas ulceradas e crostosas no plano nasal. Embora um dos animais nao apresentasse lesoes cutâneas, os sinais respiratorios eram evidentes assim como a presenca de linfadenopatia. Este estudo alerta para a ocorrencia da esporotricose em caes na regiao de Pelotas e Rio Grande, e salienta para a similaridade das lesoes observadas, com outras doencas infecciosas e neoplasicas, o que torna importante a realizacao de diagnostico diferencial.

This study aimed evaluate the antidermatophytic activity of three commercial disinfectants commonly used for environmental control of microorganisms in veterinary medicine. Sodium hypochlorite at 40 μL/mL, chloro-phenol derived at 30 μL/mL and chlorhexidine digluconate at 66.7 μL/mL were tested against 14 strains of dermatophytes, identified as Microsporum canis (n: 3) and Microsporum gypseum (n: 11). The tests was performed in accordance with guidelines of the Clinical and Laboratory Standards Institute (CLSI), documents M38-A2 and M51-A, adapted to disinfectants. In the microdilution broth test, chlorhexidine digluconate had MIC values (Minimum Inhibitory Concentration) of 4.16 μL/mL and MCF (Minimum Fungicidal Concentration) from 4.16 to 8.33 μL/mL, while chloro-phenol derived obtained MIC and MCF of 1.87 μL/mL, and both disinfectants had fungicidal activity at concentrations below the recommended. Sodium hypochlorite obtained MIC from 10 to 80 μL/mL and MFC of 40 to 80 μL/mL, requiring at most isolates twice the recommended concentration to achieve same activity. In the disc diffusion test, the mean inhibition zones for chlorhexidine digluconate was 10.53 mm, for chloro-phenol of 9.9 mm and for sodium hypochlorite was 6.2 mm. Chlorhexidine digluconate and chloro-phenol presented a significant reduction in the growth of dermatophytes, while sodium hypochlorite in concentration recommended showed a low antifungal activity against tested isolates.

Rio Grande do Sul (RS) is the second highest state with respect to sporotrichosis incidence in Brazil, with most cases occurring in the southern region. Given the importance of epidemiologic monitoring in hyperendemic areas, this study evaluated the spatial, geographical and annual sporotrichosis incidence over a period of 7 years in the southern region of RS, as well as the disease evolution over the last two decades. Data were collected from the Mycology Laboratory of the Federal University of Rio Grande (FAMED-FURG) and from the Centre for Zoonosis Control (CZC) of the Prefeitura Municipal of Pelotas city. All feline cases of sporotrichosis diagnosed between 2010 and 2016 were included and analysed. In addition, cases of human and canine sporotrichosis were accounted for. Over the 7-year period (2010-2016), 372 feline, 34 canine and 83 human cases of sporotrichosis were diagnosed, being the mean number of cases/year 18.33 in the first 3 years of the study and 116.33 in the last 3 years. Cases were distributed among 34 and 28 neighbourhoods in Rio Grande and Pelotas city, respectively. Socioeconomic features reinforced the result that the disease is already spreading across a major part of the southern region. In addition, the study demonstrated the intensification and expansion of the high endemicity areas. Therefore, given that the number of feline sporotrichosis cases in southern RS reached alarming proportions, we suggested that this region will promptly face an epidemic of sporotrichosis if no preventive or control measures are undertaken.

Experiments were conducted to evaluate the in vitro antifungal activity and determine chemical composition of Origanum vulgare essential oils. Nine Candida albicans (five clinical animal isolates and four ATCC strains), and three non-albicans species were studied; broth microdilution technique (CLSI) was used. Eight samples of commercial O. vulgare were submitted to hydrodistillation (Clevenger), and essential oils obtained were analyzed by gas chromatograph. Values of minimum inhibitory concentration (MIC) ranged from 1.0 to 0.065%, the average of MIC50 was 0.62 µL/mL while the average of MIC90 was 5.52 µL/mL and essential oils EO1 and EO8 were the ones with the best results. Results of thymol/carvacrol ratio were, for essential oils (EO) (EO1, 10.20/12.67; EO2, 7.88/0.50; EO3, 6.18/0.71; EO4, 0.58/21.58; EO5, 20.62/1.34; EO6, 21.95/4.71; EO7, 16.68/0.77; EO8, 8.42/9.44), demonstrated that differences between concentrations of thymol and carvacrol influenced MIC. These results indicate that the O. vulgare essential oils evaluated in this study presented growth inhibition activity over Candida spp. Key words: Origanum vulgare, Candida spp., essential oil, thymol, carvacrol.

The oral microbiota of humans and animals is made up of a wide variety of yeasts and bacteria, but microbiota of dogs is not totally described. Although such identification is an important step to establish the etiopathogenesis and adequate therapy for the periodontal disease The aim of this study was to evaluate and correlate oral alterations with the presence of yeasts in oral cavity of female dogs. After clinical evaluation samples from healthy and from dogs with oral diseases were obtained from three different oral sites by swabs, curettes, millimeter periodontal probes and HA membrane tip in cellulose ester. Yeast identification was performed through macroscopic and microscopic colony features and biochemical tests. Dental calculus was the most prevalent occurrence in the oral cavity of 59 females. However, the isolation of yeasts was significantly higher (p < 0.05) in animals suffering from halitosis. Eleven yeast species were identified, namely: Malassezia pachydermatis, Rhodotorula spp., Candida albicans, C. catenulata, C. famata, C. guilliermondii, C. parapsilosis, C. intermedia, Trichosporon asahii, T. mucoides and Cryptococcus albidus. It could be concluded that the yeasts are part of the microbiota from the different sites of the oral cavity of the female canines studied without causing any significant alterations except halitosis.

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.

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.

Sporotrichosis is a zoonotic mycosis, which Sporothrix brasiliensis is the prevalent agent in cats and dogs with this disease in Brazil. Itraconazole is used for the treatment, but the emergence of resistance has encouraged the search for therapeutic alternatives in medicinal plants. The benefits properties of green tea (Camellia sinensis L. Kuntze) include antifungal activity, but there are no data in sporotrichosis. This study aimed to evaluate the in vitro anti-Sporothrix spp. activity of green tea. Infusion and decoction of dried leaves of green tea were prepared at 10% and tested through broth microdilution test (CLSI M38-A2), as well as itraconazole for veterinary use. Nine clinical isolates of S. brasiliensis from cats and dogs with sporotrichosis in South region of Rio Grande do Sul, 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-1 for both extracts), without statistic difference, but the fungicidal activity was weak or no occured (MFC90 > 50 mg ml-1 for both). In itraconazole, S. brasiliensis were sensitive at the MIC50 of 2 μg ml-1 and 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 satisfactory inhibitory activity of the infusion and decoction of green tea make its promising in the treatment of sporotrichosis and more studies are needed for its use.

Neste trabalho foram avaliadas a atividade antifúngica e composição química do óleo essencial de Rosmarinus officinalis (alecrim). O óleo essencial foi obtido, através da hidrodestilação, das partes aéreas da planta, o qual foi analisado por cromatografia gasosa com detectores de ionização em chama (GC/FID) e espectrometria de massas (GC/MS). Esse óleo foi testado em isolados de C. albicans da mucosa vaginal de fêmeas caninas, casos clínicos de candidíase em animais cepas padrões e espécies não-albicans, usando a técnica de microdiluição em caldo (CLSI-M27A2). A análise cromatográfica do óleo permitiu identificar como principais compostos cânfora, verbenona e 1,8-cineol. Os valores de CIM do óleo para os isolados padrões de C. albicans variaram entre 1,25 e 2,5 µL mL-1 e de 2,5 a 5,0 µL mL-1 para CFM, enquanto que para os isolados não-albicans observou-se CIM entre 1,25 e 5,0 µL mL-1 e CFM de 2,5 a 10,0 µL mL-1. C. albicans isolada de animais apresentou valores de 2,5 a >10,0 µL mL-1 para CIM e de 5,0 a >10,0 µL mL-1 para CFM. Os resultados demonstraram atividade fungicida e fungistática do óleo essencial de alecrim em isolados de Candida spp. provenientes de animais.

Sporotrichosis is a subcutaneous mycosis, which affects mainly small animals, and is considered an important public health disease. This paper describes the epidemiological and laboratory characteristics of 103 clinical cases of sporotrichosis diagnosed over a 10-year period in southern Brazil. The 92 cats and 11 dogs from eight municipalities in Rio Grande do Sul State developed especially the disseminated cutaneous and fixed cutaneous forms of the disease. Respiratory signs such as sneezing, serous nasal discharge and dyspnea were found in about 57% of the animals. The detection of Sporothrix schenckii in different clinical samples showed highest isolation in testicles (46.6%), oral cavity (45.2%) and conjunctival mucosa (38.1%). A differentiated histological pattern was found between the fixed cutaneous and disseminated cutaneous (DC) manifestations of the disease; well-organized granulomas of nodular distribution and various fungal structures prevailed in the DC form in cats. Melanin detection in S. schenckii cells by the Fontana-Masson technique was positive in 45.4% of the samples. The study revealed that the State of Rio Grande do Sul is an endemic sporotrichosis area and demonstrated the possibility of involvement of other pathways in the infection and spread of the disease. In addition, it emphasized the importance of laboratory tests for mycosis confirmation, especially in dogs that develop clinical manifestations without the presence of cutaneous lesions.

Using transmission electron microscopy, we studied the presence of melanin and cell wall thickness of clinical isolates of Sporothrix schenckii obtained from cats, dogs and humans as compared to reference strains. We detected differences regarding presence of the melanin among the clinical isolates of S. schenckii and a correlation between presence of melanin and cell wall thickness.

Malassezia pachydermatis e uma levedura integrante da microbiota cutânea de diversas especies animais, podendo ser o agente causador de otite externa e dermatite. Recentes trabalhos descreveram o isolamento de especies lipodependentes em caes e gatos, porem estudos sobre a importância das demais especies do genero em pequenos animais, especialmente nos felinos, sao escassos. Com o objetivo de analisar a frequencia do genero no tegumento cutâneo (TC) e meato acustico externo (MAE) de felinos, foram colhidas 228 amostras de material, sendo 152 provenientes do meato acustico externo (higido ou otopata) e 76 de tegumento cutâneo (higido ou dermatopata). Todas as amostras colhidas do MAE foram submetidas a exame direto e assim como as amostras de TC foram semeadas em placas de Petri contendo os meios de cultura agar Dixon modificado e agar Sabouraud dextrose com cicloheximida e cloranfenicol. Malassezia pachydermatis foi a unica especie fungica encontrada no meato acustico externo, sendo mais comum em felinos machos que em femeas (machos 56,25%, femeas 28,41%; p M. pachydermatis apresentou baixa frequencia de isolamento em MAE e TC de felinos higidos ou com lesoes.

Clinical cases of feline sporotrichosis, originating in the Pelotas region and diagnosed at the Laboratory of Infectious Diseases (UFPel), were studied in the period from 2002 to 2006. The animals were evaluated according to the clinical forms of the mycosis, time of lesion appearance, severity of the clinical diagnosis and evolution of cutaneous lesions throughout the treatment period. Mycological analyses, carried out through direct examination, cultivation of tissue samples and exudates of feline lesions all confirmed the diagnosis of sporotrichosis in the 15 animals under study. The cutaneous dissemination form was observed in 10 animals, of which three showed prostration, anorexia and dehydration. The zoonosis occurred in 20% of case studies, and the pet owners and one attendant at a veterinary clinic were infected, developing the fixed and disseminated cutaneous forms. The treatment of mycosis was carried out with itraconazole, 10 mg kg(-1), once a day, on 12 animals. The cure of the clinical symptoms was observed on 50% of the felines. This study demonstrates a good clinical response of felines with sporotrichosis, when they were treated itraconazole and calls the attention for the incidence of human sporotrichosis on people related to the veterinary activity as well as for pet owners.

Itraconazole is currently considered the drug of choice to treat the diverse clinical presentation of sporotrichosis. On the other hand terbinafine by virtue of its excellent in vitro activity is under comparative evaluation for its therapeutic potential for a wide range of fungal infections. In this study, our aim was to determine the in vivo efficacy of terbinafine and itraconazole on a experimental model of systemic sporotrichosis. 120 rats Wistar received an injection of 2x10³ S. schenckii cells by via the lateral tail vein. After 3 days the animals were treated with terbinafine (250mg/kg) and itraconazole (100 mg/kg) and their respective diluents. In our model, terbinafine and itraconazole were effective in reducing the number of clinical lesions and positive organ cultures. There was statistical difference between the groups treated with the antifungals in relation to the control groups (pItraconazol é atualmente considerado a droga de escolha para o tratamento das diferentes formas clínicas da esporotricose. Por outro lado a terbinafina devido a sua excelente atividade in vitro está sendo avaliada quanto ao seu potencial terapêutico frente a diversas infecções fúngicas. O objetivo deste estudo foi determinar a eficácia in vivo da terbinafina e itraconazol em um modelo de esporotricose experimental sistêmica. 120 ratos Wistar receberam uma injeção de 2x10³ células de S. schenckii pela veia lateral da cauda. Após 3 dias os animais foram tratados com terbinafina (250mg/kg) e itraconazol (100mg/kg) e os seus respectivos diluentes. No modelo experimental estudado, a terbinafina e itraconazol se mostraram efetivos reduzindo o número de sintomas clínicos e retroisolamento positivo para o agente. Houve diferenças estatísticas entre os grupos tratados com os antifúngicos em relação aos grupos controle (p

Objective This work compares the virulence between pigmented (P) and albino (A) filamentous forms of S. schenckii through experimental skin sporotrichosis. Material and methods Two original wild pigmented isolates (1 and 2) cultivated in yeast extract peptone dextrose (YEPD), were exposed to 300 erg/mm2 of UV radiation in order to produce mutant albinos. Swiss albino mice were divided in four groups (P1, A1, P2 and A2) and inoculated in the right paw pad with 0.1 ml of solution (1 × 106cells/ml) and observed during nine weeks. Results The albino groups A1 (86%) and A2 (100%) presented intense inflammatory reactions in the inoculation point (p Conclusion The results obtained in this experimental model permit the conclusion that there are virulence differences between pigmented and mutant albino isolates of S. schenckii in the development of experimental skin sporotrichosis.

O controle dos helmintos gastrintestinais em ovinos, realizado exclusivamente atraves de anti-helminticos, tem como consequencia a selecao de populacoes de helmintos resistentes a praticamente todos os farmacos disponiveis. Isso tem motivado a busca de alternativas para controle, entre as quais destacamos a utilizacao de fitoterapicos. Neste contexto, o objetivo deste estudo foi avaliar a atividade ovicida in vitro do oleo essencial e do extrato hidroalcoolico de Ocimum basilicum em nematodeos de ovinos e determinar os constituintes quimicos majoritarios do oleo essencial. Para os estudos in vitro foram utilizadas fezes de ovinos naturalmente infectados, testadas frente ao oleo essencial de O. basilicum nas concentracoes de 32 a 0,25 mg/mL e ao extrato hidroalcoolico nas concentracoes 40 a 1,25 mg/mL. Como controle positivo foi utilizado o principio ativo tiabendazol (0,025 mg/mL) e negativo a agua destilada. A analise quimica do oleo essencial determinou como constituintes majoritarios chavicol (34,68%) e linalool (27,68%). O oleo essencial apresentou atividade ovicida acima de 99% em todas as concentracoes testadas. O extrato hidroalcoolico nas concentracoes entre 40 e 20 mg/mL obteve 100% de acao, decrescendo a partir de 10 mg/mL, atingindo 9,76% de atividade na concentracao 1,25 mg/mL. Os controles, positivo e negativo, obtiveram 98,57% e 8,5%, respectivamente. O valor da concentracao letal media (CL 50 ) do extrato hidroalcoolico e oleo essencial foram de 4,797 mg/mL e 0,2123 mg/mL, respectivamente. O. basilicum apresentou acao anti-helmintica in vitro sobre ovos de nematodeos gastrintestinais de ovinos, com maior potencial para o oleo essencial, constituindo um promissor fitoterapico para estudos visando a busca de moleculas bioativas para serem utilizadas no controle dos nematodeos gastrintestinais.

As dermatopatias correspondem a maioria dos atendimentos realizados em clínicas veterinárias de pequenos animais. Dentre essas, as micoses cutâneas comumente diagnosticadas são dermatofitose, malasseziose e candidose. Além disso, os cuidados com animais de estimação cresceram auxiliados pelo surgimento de pet shops. O objetivo foi isolar e identificar leveduras e dermatófitos em instrumentos de tosa de ambientes veterinários em Pelotas, RS, Brasil. Em pet shops, consultórios e clínicas veterinárias foram coletadas amostras de rascadeiras e lâminas da máquina através de swabs estéreis e impressão em placas com ágar Sabouraud acrescido de cloranfenicol, azeite de oliva e ciclohexemida, incubadas a 25º e 37ºC, por até 15 dias. As colônias filamentosas foram identificadas conforme as características do grupo dermatófitos e as leveduras, através de método automatizado. A análise de frequência de isolamento foi realizada pelo método qui-quadrado. Foram coletados 1 hospital veterinário, 4 clínicas, 4 consultórios veterinários e 6 pet shops resultando em 150 amostras, sendo 80 da lâmina da máquina de tosa e 70 da rascadeira. O isolamento fúngico foi obtido em 58 amostras, das quais 58,6% eram da lâmina da máquina de tosa e 41,4% da rascadeira, resultando em 78 isolados leveduriformes; uma vez que não foram isolados fungos filamentosos do grupo dermatófitos. Malassezia pachydermatis foi a mais frequentemente isolada, tanto nas amostras da lâmina da máquina de tosa quanto nas da rascadeira, enquanto que, Trichosporon sp. foi isolado somente nas da rascadeira. Assim, a lâmina da máquina de tosa foi a mais contaminada, com predominância de M. pachydermatis, entretanto, nenhum dermatófito foi isolado.

Objetivou-se com este estudo avaliar a atividade antifúngica in vitro do óleo essencial de Origanum vulgare frente a isolados clínicos de Malassezia pachydermatis. As folhas secas de O. vulgare foram adquiridas de distribuidor comercial com certificado de qualidade e origem e encaminhadas para extração do óleo essencial e cromatografia. Para realização do teste in vitro, foi utilizada a técnica de microdiluição em caldo (CLSI M27A3) com modificações para fitofármacos e M. pachydermatis. O óleo essencial de orégano foi testado nas concentrações de 28 a 0,87mg/mL diluído em caldo Sabouraud com 1% de tween 80. Todos os isolados foram testados em duplicata. Na análise cromatográfica do óleo essencial, foram identificados 12 compostos, sendo timol, -terpineno e 4-terpineol os compostos majoritários. A CIM e a CFM dos 42 isolados de M. pachydermatis variaram de ≤0,87 a 7mg/mL, com valores de CIM50 e CIM90 de 1,18 e 3,28mg/mL, respectivamente. Com este estudo foi possível concluir que M. pachydermatis é sensível ao óleo essencial de orégano mesmo em concentrações baixas. Dessa maneira, o óleo essencial de orégano apresenta-se como promissor na bioprospecção de novos fármacos para o tratamento das otites e dermatites na clínica de pequenos animais. The aim of this study was to evaluate the in vitro antifungal activity of essential oil of Origanum vulgare against clinical isolates of Malassezia pachydermatis. The dried leaves of O. vulgare were purchased from a commercial distributor with certified quality and origin and referred for essential oil extraction and chromatography. The technique for in vitro testing was microdilution (CLSI M27A3) with modifications to phytochemicals and M. pachydermatis. The essential oil of O. vulgare was tested at concentrations from 28 to 0.87mg/mL in Sabouraud broth diluted with 1% of tween 80. All isolates were tested in duplicate. In the chromatographic analysis of the essential oil 12 compounds were identified, and thymol, α-terpinene, 4-terpineol were the major compounds. The MIC and the MFC of the 42 isolates of M. pachydermatis ranged from ≤0.87 to 7mg/mL with MIC50 and MIC90 values of 1.18 and 3.28 mg/mL, respectively. With this study it was concluded that M. pachydermatis is sensible to O. vulgare essential oil even at low concentrations. Thus, the essential oil of O. vulgare is presented as bioprospecting in the promising new drugs for the treatment of otitis and dermatitis in small animal clinic.

The purpose of this article was to briefly review about dermatophytosis in small animals. In the last years, the pet population has increased as well the interest in having animals as pets. Because of this, these small animals are more and more inserted in our daily life. The understanding of ringworm epidemiology, diagnosis, treatment and control in pets is very important for the reason that pets are often asymptomatic carriers of dermatophytes being important sources of infection and/or carriers of infection. So, it is very important to know about this disease to reduce the spread of zoophilic fungal infections in humans, considering the close contact between pets and their owners, especially between children and their pets.

Due to anthelmintic resistance in nematodes, several research studies have been developed seeking control alternatives to these parasites. This study evaluated the in vitro action of Origanum vulgare on gastrointestinal nematode eggs of cattle. In order to evaluate the ability to inhibit egg hatch, different dried leaves extracts of this plant were tested, such as dye, hydroalcoholic and aqueous extracts at concentrations varying from 0.62 to 80 mg/mL. Each assay was accompanied by control containing levamisole hydrochloride (0.2 mg/mL), distilled water and 70o GL grain alcohol at the same concentration of the extracts. Test results showed that the different O. vulgare extracts inhibited egg hatch of cattle gastrointestinal nematodes at a percentage that varied from 8.8 to 100%; dye and hydroalcoholic extract were the most promising inhibitors. In view of this ovicidal property, O. vulgare may be an important source of viable antiparasitic compounds for nematodiosis control in ruminants. KEYWORDS: Haemonchus spp.; in vitro; oregano; phytotherapic.

Programas de higiene e desinfecção são essenciais no controle ambiental de agentes fúngicos potencialmente patogênicos aos homens e animais, dessa forma objetivou-se avaliar a ação antifúngica do hipoclorito de sódio 4% e digluconato de clorexidina 6,6% em doze isolados fúngicos. Foram testados fungos isolados do ambiente e de casos clínicos como Aspergillus spp, Candida spp, Microsporum spp, Malassezia pachydermatis, Cryptococcus neoformans e Sporothrix schenckii. Foram utilizadas as técnicas de microdiluição em caldo e difusão em ágar conforme protocolos descritos pelo Clinical Laboratory Standard Institute (CLSI) com adaptações para agentes químicos. Todos os isolados foram sensíveis a concentrações inferiores a recomendada pelo fabricante do digluconato de clorexidina (6,6%) com CIM e CFM entre ≤0,42 a 1,68%. Para o hipoclorito de sódio, 58,3% dos isolados foram resistentes a concentração recomendada pelo fabricante (4%) apresentando CIM e CFM entre 2 e >8%. O digluconato de clorexidina apresentou maiores zonas de inibição do que o hipoclorito de sódio, com halos de até 31mm de diâmetro. A clorexidina demonstrou ação fungicida em baixas concentrações sendo eficaz na eliminação de fungos filamentosos e leveduriformes de importância médica e veterinária, enquanto o hipoclorito de sódio somente obteve esta ação em altas concentrações.

Fungi of the Aspergillus genus are widespread and contaminate the environment. Thousands of conidia are released from each phialide and dispersed in the air every day. These fungi are considered important mycose-causing agents in hospitals. Due to this, research to determine prevalent fungi from the Aspergillus genus in hospital environments, and an adequate disinfection program in these areas is are needed. This study evaluated the susceptibility of Aspergillus spp. isolated from a veterinary environment against four disinfectants. Successive dilutions of disinfectants (log2) were used according to CLSI M38-A2 microdilution technique adapted to chemical agents against 18 isolates of this genus. After 72 hours of incubation, the Minimum Inhibiting Concentration and Minimum Fungicidal Concentration capable of inhibiting 50% and 90% of the isolates were determined. Chlorexidine-cetrimine, benzalconium chloride and a chlorophenol derivative proved to be effective against all isolates with a lower MIC than that suggested by the manufacturer, except for the A. flavus strain. Sodium hypochlorite was ineffective against three A. fumigatus, three A. flavus and one A. niger isolate. These results demonstrated that all studied disinfectants were effective against environmental isolates, with the exception of sodium hypochlorite, which showed lower effectiveness.

The literature shows that is a resistance by the microorganisms against commercial products used for the disinfection of contaminated environments. Due to this, the aim of this study was to evaluate the activity of the tincture, aqueous extract (decoction and infusion) and hydroalcoholic extract of Origanum vulgare and Rosmarinus officinalis, as an alternative to disinfectants, using the technique of direct exposure, against Sporothrix schenckii complex. For this, the direct exposure tests, with fungal isolates from clinical cases pertaining to dogs and cats with sporotrichosis, were used to evaluate the sensitivity of Sporothrix spp. against plants extracts, with the action evaluated in the times of 5, 10 and 15 minutes of exposure . As a positive control, it was used a sterile water solution containing innocuous and disinfectants used to control was sodium hypochlorite alcohol and 70°GL. As a result, it can be shown the sensitivity of the fungal isolates front to tincture of Origanum vulgare and Rosmarinus officinalis and hydroalcoholic extract of Origanum vulgare at all times of exposure, in 10% concentration of the extract.

Relatou-se um caso clínico de dermatite multifatorial em cão, envolvendo Demodex canis, Sporothrix schenckii e bactérias dos gêneros Staphylococcus e Streptococcus. O animal, macho, da raça Pit Bull, com um ano e três meses de idade, apresentou lesões úmidas, hiperêmicas, descamativas e ulcerativas nas regiões do pescoço e do tórax. Exames laboratoriais confirmaram o diagnóstico de infecção concomitante por fungo, bactéria e parasito. A terapia antimicrobiana com moxidectina, itraconazol e amoxicilina foi efetiva, com remissão dos sinais clínicos após 30 dias de tratamento. Este relato salienta a importância do diagnóstico de certeza, que foi determinante para estabelecimento da terapêutica adequada e recuperação do animal. A clinical case of dermatitis multifactorial in a dog was reported, involving Demodex canis, Sporothrix schenckii, and bacteria of the genera Staphylococcus and Streptococcus. The animal, male, Pit Bull, with a year and three months of age had soaked lesions, was hyperemic, scaly and had an ulcer in the neck and chest. Laboratory tests confirmed the diagnosis of concomitant infection by fungi, bacteria and parasites. Antimicrobial therapy with moxidectin, itraconazol and amoxicillin was effective, with remission of clinical signs after 30 days of treatment. This report emphasizes the importance of an accurate diagnosis, which was crucial to establish the correct treatment and recovery of the animal.

This study describes the isolation of S. schenckii in hospital and home environments in Brazil. Samples were collected from surfaces of a veterinary service place and at home. S. schenckii was detected in 1.5% of the samples from the hospital environment. However, this fungus was isolated from all sampled areas in home environments. The isolation of S. schenckii deonstrates that these surfaces could act as infection sources to anials and huans. Therefore, employees and pet owners could be exposed to this agent, and the contamination, through surfaces, could occur through the traumatic inoculation of the fungus or by direct contact with pre-existing lesions.Esse estudo descreve o isolamento de S. schenckii em ambiente hospitalar e domiciliar, no Brasil. Foram colhidas amostras de superfície de local de atendimento veterinário e ambiente domiciliar. S. schenckii foi isolado em 1,5% das amostras do ambiente hospitalar. Entretanto, esse fungo foi isolado em todas as amostras do ambiente domiciliar. O isolamento do S. schenckii demonstra a importância dessas superfícies atuarem como fontes de infecção para animais e humanos. Portanto, funcionários e proprietários de animais de estimação estariam expostos a esse agente e a contaminação, através das superfícies, poderia ocorrer pela inoculação traumática do fungo ou pelo contato direto com lesões pré-existentes.

The susceptibility of Sporothrix schenckii isolates from clinical cases of canine, feline and human sporotrichosis, and from the environment, was evaluated with 4% sodium hypochlorite and 6.6% chlorhexidine digluconate using the broth microdilution, agar diffusion and direct exposure techniques. The minimal inhibitory concentration was smaller than 0.8% for chlorhexidine digluconate and between 8% and 4% for sodium hypochlorite. Inhibition zones were not found in agar diffusion for sodium hypochlorite, and zones averaging 1.9 mm were found for chlorhexidine digluconate. In the direct exposure test, sodium hypochlorite demonstrated best performance at 20 min of contact, as chlorhexidine digluconate presented little antimicrobial activity.

Background Dermatophytic pseudomycetoma is a subcutaneous fungal infection by Microsporum canis . Aims This work describes a case of dermatophytic pseudomycetoma in a Persian cat. Patient and methods A 3-year old female Persian cat showing alopecia, scaling and ulcerated nodules throughout the body, with presence of ulcerated nodules with yellow granular discharges on the dorsum, close to the tail. Mycological and histopathological examinations were realized. Results Diagnosis of dermatophytic pseudomycetoma was established. The cat was treated with griseofulvin, and surgical excision was carried out. Response to therapy was effective during the first months, during which a reduction in nodule frequency was observed. However, despite maintaining the therapy levels, the lesions relapsed and progressed to the point of causing the animal's death. Conclusion The clinic evolution showed the progressive and recurrent character of dermatophytic pseudomycetoma.

Espécies fúngicas do gênero Aspergillus são frequentemente associadas com alta mortalidade de aves marinhas em cativeiro. Tendo em vista que a aspergilose geralmente é adquirida pela inalação dos propágulos fúngicos presentes no ar, o estudo objetivou avaliar a qualidade do ar quanto às espécies de Aspergillus, das instalações internas de um centro de reabilitação de animais marinhos que frequentemente recebe pingüins, gaivotas, albatrozes e petréis acometidos por alguma moléstia. Oitenta e um dias de colheitas foram realizados distribuídos em um período de aproximadamente dois anos, através da exposição de placas de Petri contendo Agar Sabouraud dextrose acrescido de cloranfenicol no ambiente, as quais foram posteriormente incubadas a 25ºC. As colônias identificadas como pertencentes ao gênero Aspergillus, foram classificadas quanto à espécie através da avaliação macro e micro morfológica associada a uma chave de identificação. Foram obtidos 43 isolados classificados em sete espécies distintas, sendo A. fumigatus a predominante correspondendo a 27,9%, seguida de A. niger, A. flavus e outras quatro espécies de Aspergillus sp., demonstrando que as aves marinhas estão expostas a espécies fúngicas com potencial patogênico, o que enfatiza a necessidade de um controle microbiológico no ambiente onde são mantidos os animais em cativeiro. Aspergillosis in captivity seabirds is often associated with elevated rates of mortality. The infection is usually acquired by inhalation of airborne fungal conidia. The aim of this study was to evaluate the presence of Aspergillus species in the indoor environment of a rehabilitation centre for marine animals in Southern Brazil. This centre continuously receives injured penguins, seagulls, albatrosses and petrels. Petri dishes plates with Agar Sabouraud dextrose and chloramphenicol were left open for 15 minutes in 3 distinct points in the rehabilitation centre and then incubated at 25ºC. During a period of two years the indoor air was sampled in 81 occasions. A total of 43 isolates belonging to 7 different Aspergillus species were recovered. Aspegillus fumigatus was the predominant species (27.9%), followed by A. niger (25.6%), and A. flavus (16.3%). Four other Aspergillus species were isolated. This study demonstrates that seabirds were exposed to pathogenic Aspergillus species in our rehabilitation centre, reinforcing the need for a strict microbiology control of the indoor air in the captivity environment.

Itraconazole is currently considered the drug of choice to treat the diverse clinical presentation of sporotrichosis. On the other hand terbinafine by virtue of its excellent in vitro activity is under comparative evaluation for its therapeutic potential for a wide range of fungal infections. In this study, our aim was to determine the in vivo efficacy of terbinafine and itraconazole on a experimental model of systemic sporotrichosis. 120 rats Wistar received an injection of 2x10³ S. schenckii cells by via the lateral tail vein. After 3 days the animals were treated with terbinafine (250mg/kg) and itraconazole (100 mg/kg) and their respective diluents. In our model, terbinafine and itraconazole were effective in reducing the number of clinical lesions and positive organ cultures. There was statistical difference between the groups treated with the antifungals in relation to the control groups (p

Os ambientes hospitalares podem albergar micro-organismos patogênicos e oportunistas, capazes de infectar, particularmente, os indivíduos imunossuprimidos. O processo de limpeza e desinfecção assume importância capital para o controle de tais doenças. O presente trabalho avaliou a suscetibilidade de leveduras isoladas de superfícies de ambiente hospitalar veterinário frente a quatro desinfetantes/antissépticos comumente utilizados na rotina de desinfecção. O teste de microdiluição em caldo foi realizado com 24 isolados leveduriformes frente ao hipoclorito de sódio 4%, cloreto de benzalcônico 2%, clorexidina-cetrimida 6,6% e derivado de cloro-fenol 3%. A clorexidina-cetrimida, cloreto de benzalcônio e derivado de cloro-fenol foram eficazes em todos os isolados com concentração inibitória mínima e concentração fungicida inferiores à concentração recomendada pelo fabricante. Por outro lado, a ação fungicida do hipoclorito de sódio em 79,1% dos isolados testados foi obtida na concentração recomendada pelo fabricante, com desempenho inferior aos demais desinfetantes avaliados.