scholarly journals Avaliação do uso da resazurina em teste de suscetibilidade in vitro frente a Sporothrix brasiliensis

2019 ◽  
Vol 31 (2) ◽  
pp. 32-37
Author(s):  
Beatriz Mendes Roca ◽  
Vanice Rodrigues Poester ◽  
Antonella Souza Mattei ◽  
Gabriel Baracy Klafke ◽  
Ivy Bastos Ramis ◽  
...  

A técnica de microdiluição em caldo, que permite determinar a concentração inibitória mínima (CIM) dos fármacos frente aos microrganismos, é considerada padrão ouro para avaliação do perfil de suscetibilidade fúngica in vitro. Considerando que a leitura dos resultados é realizada pela visualização do crescimento fúngico, e que a utilização de indicadores de crescimento microbiano poderia otimizar essa leitura, o estudo teve como objetivo avaliar o uso da resazurina como indicador do crescimento de Sporotrhix brasiliensis em testes de microdiluição em caldo. Cinco isolados clínicos (nas fases leveduriforme e filamentosa) foram testados frente ao itraconazol (ITC – concentrações de 0,0313 a 16 µg/mL) a partir da técnica de microdiluição em caldo, utilizando a resazurina nas concentrações de 0,01% e 0,002%. A leitura dos resultados foi realizada após 24, 48 e 72 horas de incubação, e a CIM dos poços com resazurina foi considerada como sendo a menor concentração sem modificação na coloração, enquanto que a CIM do controle, foi considerada a menor concentração de fármaco capaz de inibir o crescimento fúngico. Os resultados da mudança da cor da resazurina foram comparados com os resultados sem acréscimo de resazurina. A CIM do ITC na microdiluição clássica frente aos isolados testados variou de 0,0625 a 1 µg/mL, porém, com o uso da resazurina como indicador de crescimento fúngico, independente da concentração testada, a CIM foi ≤0,0625 µg/mL. O estudo evidencia que a resazurina não é eficaz como indicador de crescimento de S. brasiliensis visto que não modificou a coloração do meio de cultivo, mesmo na presença visível de crescimento fúngico, sendo portanto contra indicada para uso em testes de suscetibilidade in vitro frente a este microrganismo.

2018 ◽  
Vol 18 (2) ◽  
pp. 164-171 ◽  
Author(s):  
Luana da S.M. Forezi ◽  
Luana Pereira Borba-Santos ◽  
Mariana F.C. Cardoso ◽  
Vitor F. Ferreira ◽  
Sonia Rozental ◽  
...  

Sporotrichosis is a serious public health problem in Brazil that affects human patients and domestic animals, mainly cats. Thus, the search for new antifungal agents is required also due to the emergence and to the lack of effective drugs available in the therapeutic arsenal. The aim of this study was to evaluate the in vitro antifungal profile of two synthetic series of coumarin derivatives against Sporothrix schenckii and Sporothrix brasiliensis. The three-components synthetic routes used for the preparation of coumarin derivatives have proved to be quite efficient and compounds 16 and 17 have been prepared in good yields. The inhibitory activity of nineteen synthetic coumarins derivatives 16a-i and 17a-j were evaluated against Sporothrix spp. yeasts and the most potent compounds were 16b and 17i. However, according to concentrations able to inhibit (minimum inhibitory concentrations) and kill (minimum fungicidal concentrations) the cells, 17i was more effective than 16b against Sporothrix spp. Thus, 17i exhibited good antifungal activity against S. brasiliensis and S. schenckii, suggesting that it is an important scaffold for the development of novel antifungal agents.


2022 ◽  
Vol 15 (1) ◽  
pp. 55
Author(s):  
Lais Cavalcanti dos Santos Velasco de Souza ◽  
Lucas Martins Alcântara ◽  
Pãmella Antunes de Macêdo-Sales ◽  
Nathália Faria Reis ◽  
Débora Sena de Oliveira ◽  
...  

Recently, the well-known geographically wide distribution of sporotrichosis in Brazil, combined with the difficulties of effective domestic feline treatment, has emphasized the pressing need for new therapeutic alternatives. This work considers a range of synthetic derivatives as potential antifungals against Sporothrix brasiliensis isolated from cats from the hyperendemic Brazilian region. Six S. brasiliensis isolates from the sporotrichotic lesions of itraconazole responsive or non-responsive domestic cats were studied. The minimum inhibitory concentrations (MICs) of three novel hydrazone derivatives and eleven novel quinone derivatives were determined using the broth microdilution method (M38-A2). In silico tests were also used to predict the pharmacological profile and toxicity parameters of these synthetic derivatives. MICs and MFCs ranged from 1 to >128 µg/mL. The ADMET computational analysis failed to detect toxicity while a good pharmacological predictive profile, with parameters similar to itraconazole, was obtained. Three hydrazone derivatives were particularly promising candidates as antifungal agents against itraconazole-resistant S. brasiliensis from the Brazilian hyperendemic region. Since sporotrichosis is a neglected zoonosis currently spreading in Latin America, particularly in Brazil, the present data can contribute to its future control by alternative antifungal drug design against S. brasiliensis, the most virulent and prevalent species of the hyperendemic context.


2018 ◽  
Vol 67 (3) ◽  
pp. 463-463
Author(s):  
Thalita Gagini ◽  
Luana Pereira Borba-Santos ◽  
Anderson Messias Rodrigues ◽  
Zoilo Pires de Camargo ◽  
Sonia Rozental

2016 ◽  
Vol 181 (11-12) ◽  
pp. 857-863 ◽  
Author(s):  
Stefanie Bressan Waller ◽  
Isabel Martins Madrid ◽  
Anna Luiza Silva ◽  
Luciana Laitano Dias de Castro ◽  
Marlete Brum Cleff ◽  
...  

Author(s):  
Luana Pereira Borba-Santos ◽  
Thayná Lopes Barreto ◽  
Taissa Vila ◽  
Kung Darh Chi ◽  
Fabiana dos Santos Monti ◽  
...  

Sporotrichosis has become an important zoonosis in Brazil and Sporothrix brasiliensis is the primary species transmitted by cats. Improvement of animal treatment will help control and limit the spread and geographic expansion of sporotrichosis. Accordingly, buparvaquone, an antiprotozoal hydroxynaphthoquinone agent marketed as Butalex®, was evaluated in vitro and in vivo against feline-borne isolates of S. brasiliensis . Buparvaquone inhibited in vitro fungal growth at concentrations 4-fold lower than itraconazole (the first-choice antifungal used for sporotrichosis) and was 408 times more selective for S. brasiliensis than mammalian cells. Yeasts treated with a subinhibitory concentration of buparvaquone exhibited mitochondrial dysfunction, ROS and neutral lipid accumulation, and impaired plasma membranes. Also, scanning electron microscopy images revealed buparvaquone altered cell wall integrity and induced cell disruption. I n vivo experiments in a Galleria mellonella model revealed that buparvaquone (single dose of 5 mg/kg) is more effective than itraconazole against infections with S. brasiliensis yeasts. Combined, our results indicate that buparvaquone has a great in vitro and in vivo antifungal activity against S. brasiliensis , revealing the potential application of this drug as an alternative treatment for feline sporotrichosis.


2017 ◽  
Vol 55 (8) ◽  
pp. 869-876 ◽  
Author(s):  
Karine Ortiz Sanchotene ◽  
Tchana Martinez Brandolt ◽  
Gabriel Baracy Klafke ◽  
Vanice Rodrigues Poester ◽  
Melissa Orzechowski Xavier

2017 ◽  
Vol 105 ◽  
pp. 117-121 ◽  
Author(s):  
Stefanie B. Waller ◽  
Cristina M. Peter ◽  
Jéssica F. Hoffmann ◽  
Tony Picoli ◽  
Luiza da G. Osório ◽  
...  

2016 ◽  
Vol 68 (4) ◽  
pp. 991-999 ◽  
Author(s):  
S.B. Waller ◽  
I.M. Madrid ◽  
M.B. Cleff ◽  
R. Santin ◽  
R.A. Freitag ◽  
...  

ABSTRACT 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.


2019 ◽  
Vol 71 (3) ◽  
pp. 819-827
Author(s):  
C.M. Peter ◽  
S.B. Waller ◽  
T. Picoli ◽  
L.G. Osório ◽  
J.L. Zani ◽  
...  

ABSTRACT In this study, we described the antifungal activity of three Brazilian propolis extracts: brown, green and from jataí bees against Sporothrix brasiliensis. The extracts were obtained from ethanolic extraction and their chemical composition was determined by high-performance liquid chromatography coupled to mass spectrometry. The cellular toxicity was measured in MDBK (Madin-Darby Bovine Kidney) cells and quantified by the MTT assay (3- (4,5 dimethylthiazol-2yl -2,5-diphenyl-2H bromine tetrazolato). For antifungal activity, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by broth microdilution. The results showed that cell toxicity was not observed at lower concentrations (0.097 to 0.39μg/ml) for all extracts in comparison to cell control. Among the chemical compounds identified, caffeic acid, p-coumaric acid, chlorogenic acid, ferulic acid and rutin were quantified. In antifungal activity, green and jataí did not exhibit activity against the isolates (MIC and MFC greater than 0.78mg/ml). However, all isolates of S. brasiliensis were sensitive to brown propolis (MIC of 0.09 to 0.78mg/ml), including the standard strain (P<0.001). Among the Brazilian propolis studied, the brown propolis showed activity against the S. brasiliensis isolates and more studies should be undertaken in order to evaluate its promising use in the treatment of sporotrichosis.


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