Atividade in vitro do óleo essencial de Origanum vulgare L. em isolados clínicos de Aspergillus spp / In vitro activity of essential oil of Origanum vulgare L. in clinical isolates from Aspergillus spp

Aspergillus fumigatus, A. flavus e A. niger são os mais importantes agentes etiológicos da aspergilose, relevante micose aviária, com tratamento ineficaz e altas taxas de mortalidade. Em vista da importância da aspergilose, da necessidade de prospectar novos fármacos e do potencial terapêutico do óleo essencial de Origanum vulgare L. (OEO), o orégano, objetivou-se avaliar a sensibilidade in vitro de isolados clínicos de Aspergillus spp. em relação ao OEO. O óleo foi obtido por hidrodestilação em Clevenger, e a análise química realizada por cromatografia de massa (GC/MS). Observaram-se 15 diferentes compostos ativos, sendo 4-terpineol, hidrato de sabinene e timol os majoritários. Nos testes de microdiluição em caldo (Reference..., 2008), todos os isolados (n= 23) foram sensíveis ao OEO: A. fumigatus teve CIM entre 28,125mg/mL (0,1875%) e 450mg/mL (3,0%), A. flavus entre 112,5mg/mL (0,75%) e 450mg/mL, e A. niger 112,5mg/mL. CFM variou de 112,5mg/mL a 450mg/mL nos isolados de A. fumigatus, de 225mg/mL (1,5%) a 450mg/mL em A. flavus, e foi de 450mg/mL em A. niger. CIM e CFM foram idênticos em 6/14 isolados, o que demonstra que o óleo com a mesma concentração pode ter capacidade fungistática e fungicida. CIM 90 correspondeu à CIM máxima. Os resultados demonstram a atividade anti-Aspergillus do OEO, com CIM 90 de 450mg/mL (3%).(AU)

Aspergillus fumigatus, A. flavus and A. niger are the most important etiological agents of aspergillosis, a relevant avian mycosis, with innefective treatment and high mortality rates. Due the importance of aspergillosis, the necessity of prospection of new drugs and the therapeutic potential of the essential oil of Origanum vulgare L. (OEO), popularly known as oregano, aims to evaluate the in vitro sensitivity of Aspergillus spp. opposing to OEO. The oil was obtained by hydrodistillation in Clevenger, and the chemical analysis performed by mass chromatography (GC/MS). 15 different active compounds were observed, being 4-terpineol (18.4%), sabinene hydrate (15.6%) and thymol (13.6%), the majority components. In the in vitro susceptibility test, all strains showed sensitivity to OEO, MIC of Aspergillus fumigatus ranged from 28,125mg/mL (0,1875%) to 450mg/mL (3,0%), A. flavus 112,5mg/mL (0,75%) to 450mg/mL, and A. niger 112,5mg/mL. MFC ranged from 112,5mg/mL to 450mg/mL in the A. fumigatus isolates, 225mg/mL (1,5%) to 450mg/mL in A. flavus, and 450mg/mL for A. niger. The MIC and FMC values were identical in 6/14 of the isolated subjects, demonstrating that the oil with the same concentration can have fungistatic and fungicidal capacity. The results demonstrates anti-Aspergillus activities of OEO with CIM90 de 450mg/mL (3%).(AU)

Eliminação de Dioctophyme renale pela urina em canino com dioctofimatose em rim esquerdo e cavidade abdominal - Primeiro relato no Rio Grande do Sul / Elimination of Dioctophyme renale to urine in canines with dioctophymosis in the left kidney and abdominal cavity - First report in Rio Grande do Sul

Dioctophyme renale é um parasito que afeta tanto animais como humanos e tem como órgão de eleição o rim direito. Relata-se o caso clínico-cirúrgico de um paciente canino com histórico de eliminação de três exemplares de D. renale pela urina e presença de parasitos no rim esquerdo e na cavidade abdominal. No Hospital de Clínicas Veterinárias da Universidade Federal de Pelotas, foram realizados exames pré-cirúrgicos, entre eles a ultrassonografia abdominal, que identificou a localização dos nematódeos, e o exame de Doppler pulsado dos vasos intrarrenais, que demonstrou aumento nos índices resistivos nas regiões avaliadas. O paciente foi conduzido para a realização de laparotomia exploratória, sendo removidos 23 parasitos da cavidade abdominal. A presença de D. renale causa graves lesões nos rins e nos demais órgãos da cavidade abdominal, sendo essencial o diagnóstico precoce e a remoção dos nematódeos para recuperação do paciente.(AU)

Dioctophyme renale is a parasite that attacks animals as well as humans and has the right kidney as a preferred target organ. This document reports the case of a clinical-chirurgical canine patient with a history of elimination of three D. renale samples through its urine and presence of parasites at its left kidney and abdominal cavity. Pre-cirurgical exams were performed at the Veterinary Hospital of the Federal University of Pelotas. Among these exams, abdominal ultrasonography identified the nematode's location and the Pulse-Doppler of the intrarenal vessels showed an increase in the resistive indices of the evaluated areas. An Exploratory Laparotomy was conducted with the patient who had 23 parasites removed from its abdominal cavity. The presence of D. renale causes severe lesions at kidneys and other organs of the abdominal cavity, for this reason early diagnosis and nematode´s removal are essential for the patient's recovery.(AU)

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

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

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

Levantamento de extratos vegetais com ação anti-Candida no período de 2005-2013 / Survey of plant extracts with anti-Candida activity in the 2005-2013 period

Entre as micoses relevantes em saúde pública destaca-se a candidíase, infecção oportunista que acomete o homem e animais. A enfermidade era considerada pouco frequente na medicina veterinária, porém relatos demonstram um aumento considerável, assim como a resistência aos antifúngicos. Com isso, pesquisas têm sido desenvolvidas visando encontrar substâncias bioativas frente ao gênero Candida. Desta forma, objetivou-se reunir dados das bases Scielo e ScienceDirect com informações entre os anos de 2005-2013 referentes à ação anti-Candida de diferentes extratos vegetais. Foi encontrado um total de 78 famílias e 208 espécies de plantas com atividade frente à Candida spp., destacando-se as famílias Asteraceae, Geraniaceae, Myrtaceae, Fabaceae, Lamiaceae, Rubiaceae, Verbenaceae e Anacardiaceae, com extratos diclorometânicos, aquosos, etanólicos, metanólico, frações e subfrações, sendo as folhas a parte vegetal mais utilizada. As plantas descritas apresentaram ação anti-Candida, porém algumas necessitam concentrações muito altas dos extratos com pequena inibição de crescimento/eliminação destas leveduras, ocorrendo variações, principalmente, quanto ao método de avaliação, tipo de extrato, parte vegetal, e procedência dos isolados fúngicos. Chama a atenção a raridade dos estudos com isolados de animais, principalmente de casos clínicos. Por fim, destacam-se as famílias Asteraceae e Geraniaceae que apresentaram maior número de espécies vegetais com atividade, podendo ser uma fonte de investigação frente à Candida spp.

Among the relevant mycoses in public health, one that stands out is candidiasis, an opportunistic infection that affects humans and animals. The disease was considered uncommon in veterinary medicine, but reports show a significant increase, as well as resistance, to conventional antifungal agents. Therefore, research has been undertaken aimed at finding bioactive substances from plants that fight against Candida. Thus, the objective of this work was to gather the databases SciELO and ScienceDirect with information between the years 2005 and 2013 concerning the anti-Candida activity of different plant extracts. A total of 78 families and 208 species of plants with activity against Candida spp. was found highlighting the Asteraceae, Geraniaceae, Myrtaceae, Fabaceae, Lamiaceae, Rubiaceae, Verbenaceae and Anacardiaceae families, with dichloromethane, aqueous, ethanol and methanol extracts and fractions and subfractions, being the leaf the most used plant part. The plants described showed anti-Candida activity, but some require very high concentrations of the extracts with little growth inhibition / elimination of these yeasts, with variations related mainly to the method of assessment, type of extract, plant parts and origin of the fungal isolates. The rarity of studies with isolates from animals, mainly clinical cases, draws attention. Finally, we highlight the Asteraceae and Geraniaceae families, which had a greater number of plant species with activity and which may be a source of research against Candida spp.

Atividade antifúngica do óleo essencial de Origanum vulgare frente a Malassezia pachydermatis / Antifungal activity of Origanum vulgare essential oil against Malassezia pachydermatis

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, a-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...

Dermatite multifatorial em um canino / Multifactorial dermatitis in a dog

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.

Perfil de suscetibilidade de leveduras do gênero Candida isoladas de animais ao óleo essencial de Rosmarinus officinalis L / Susceptibility profile of Candida spp. isolated from animals to the essential oil of Rosmarinus officinalis L

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.

In this work the antifungal activity and chemical composition of the Rosmarinus officinalis (rosemary) essential oil against Candida spp. were evaluated. The essential oil was obtained by hydrodistillation of aerial parts, and analyzed by gas chromatography with a flame ionization detector (GC/FID) and gas chromatography coupled with mass spectrometry (GC/MS). This oil was tested against fourteen isolates of Candida sp. (ten C. albicans and four no-albicans species) by the microdillution broth assay (CLSI-M27A2). Chromatography analyses of the essential oil showed compounds, as camphor, verbenone and 1,8-cineole as major constituents. MIC and MFC values for C. albicans standard strains were 1.25 - 2.5 µL mL-1 and 2.5 - 5.0 µL mL-1, respectively. MIC and MFC for the non-albicans species were 1.25 - 5.0 µL mL-1 and 2.5 - 10.0 µL mL-1, respectively. C. albicans isolates obtained from animals exhibited MIC and MFC values of 2.5 - > 10.0 µL mL-1 and 5.0 - > 10.0 µL mL-1, respectively. According to the results, the essential oil of rosemary presented fungicidal and fungistatic activities against Candida spp.

Atividade inibitória do óleo essencial de orégano em fungos de importância médica e veterinária / Inhibitory activity of origanum essential oil against important fungus in veterinary

The in vitro activity of Origanum vulgare essential oil against fungal isolates was evaluated. A total of 27 clinical isolates were used, including: C. albicans, S. schenckii, M. pachydermatis, Aspergillus flavus, and A. fumigatus. Microdilution in broth technique (NCCLS M27-A2 and M-38) was used and susceptibility was expressed as Minimum Inhibitory Concentration (MIC). Essential oil was obtained by hydrodistillation in Clevenger and analyzed by gas chromatography, showing the presence of 4-terpineol, alpha-terpineol, 4-terpinene, thymol and carvacrol, as the main compounds. Origanum oil MIC for C. albicans varied from 125 to 500mL/mL; for S. schenckii, from 250 to 500mL/mL; for M. pachydermatis, from 15 to 30mL/mL; and for Aspergillus, from 30 to 60mL/mL. Isolates sensitivity showed to the origanum oil stimulates the accomplishment of new studies, including in vivo tests, contributing to the search of alternative treatments to mycosis.

Eficácia da clorexidina-cetrimida na desinfecção ambiental contra Aspergillus spp. / Chlorexidine-cetrimide efficacy in ambient disinfection against Aspergillus spp

Avaliou-se a ação in loco da clorexidina-cetrimida no controle de Aspergillus spp., considerando-se a influência de fatores climáticos e populacionais, em um centro de recuperação de animais marinhos. Durante dois anos, realizaram-se colheitas de amostras de ar por meio da técnica de sedimentação, com implantação no segundo ano de um programa de desinfecção com clorexidina-cetrimida. Os resultados do isolamento fúngico nos dois anos foram comparados estatisticamente pelo programa Epinfo 8.0. Demonstrou-se que o isolamento de Aspergillus spp. não apresentou relação significativa com fatores climáticos e populacionais e que a diminuição da concentração de conídios de Aspergillus spp. do ambiente ocorreu devido ao programa eficaz de desinfecção com a clorexidina-cetrimida.

The study evaluated the in loco activity of chlorexidine-cetrimide for environmental control of Aspergillus spp. considering the climatic influence and population variations, in a recuperation center of marine animals. Samples of air were collected throughout two years by the sediment method, and the disinfection program was established in the second year. Data collected during the two years were statistically compared by Epinfo 8.0. program. It was demonstrated that both population and climate had no influence on the fungal isolation, and the disinfection program with chlorexidine-cetrimide was considered effective in the reduction of conidia Aspergillus concentration indoor.

Esporotricose experimental sistêmica em ratos Wistar: avaliação hematológica e perfil hepático / Systemic experimental sporotrichosis in Wistar rats: hematological evaluation and hepatic profile

Alanine aminotransferase (ALT) and alkaline phosphatase (AP) hepatic enzymes, hematological values, and total plasmatic proteins (TPP) of 12 Wistar rats with systemic experimental sporotrichosis were evaluated. Inoculation was performed at 2x10³ cells of S. schenckii/ml. The values ranged from 35 to 48UI/l for ALT and 67 to 251.3UI/l for AP. Fifty percent of the AP values were above the physiological limits. TPP was elevated in 66.7 percent (8/12) of the rats with values varying from 7.9 to 8.5g percent. In 75 percent (9/12) of the animals, it was observed that hemoglobin and red blood cells count were below the physiologic limits, and 8.3 percent (1/12) of the animals showed anemia masked by dehydration. At the necropsy, it was observed local and/or disseminated nodular lesions. At the histological analysis, cells compatible to S. schenckii were found. The observed alterations are aggravating factors for the recurrence and treatment of the mycosis.

Atividade in vitro do óleo essencial de Origanum vulgare frente à Sporothrix Schenckii / In vitro activity of the essential oil of Origanum vulgare against Sporothrix schenckii

In vitro activity of the essential oil Origanum vulgare against Sporothrix schenckii was determined by the MIC (Minimum Inhibitory Concentration). For this, seven samples of S. schenckii were studied, two isolated from two cases of human sporotrichosis and five isolated from cats. Analysis of the essential oil was carried out in a gas chromatograph (GC/FID) for the identification and quantification of thymol and carvacrol (antifungal agents). MIC was obtained based on the microdilution method according to the adapted document NCCLS-M 27A2 for fitopharmacy. All the isolates presented sensibility to the essential oil. S. schenckii was inhibited in a concentration of 0.25 percent (250m l/ml). Chromatographic analysis showed that thymol concentration was bigger than carvacrol. The antifungal activity demonstrated by the essential oil of O. vulgare against S. Schenckii stimulates the accomplishment of more studies, including in vivo studies.