RESUMO
Microorganisms adapt to environmental conditions as a survival strategy for different interactions with the environment. The adaptive capacity of fungi allows them to cause disease at various sites of infection in humans. In this study, we propose digital images as responses of a complete factorial 23. Furthermore, we compared two experimental approaches: the experimental design (3D) and the checkerboard assay (2D) to know the influence of pH, glucose, and fluconazole concentration on different strains of the genus Candida. The digital images obtained from the factorial 23 were used as input in the PCA-ANOVA to analyze the results of this experimental design. pH modification in the culture medium modifies the susceptibility in some species less adapted to this type of modification. For the first time, to the best of our knowledge, digital images were used as input to PCA-ANOVA to obtain information on Candida spp.. Therefore, a higher concentration of antifungals is needed to inhibit the same strain at a lower pH. In short, we present an alternative with less use of reagents and time. In addition, the use of digital images allows obtaining information about fungal susceptibility with three or more factors.
Assuntos
Candida , Glucose , Antifúngicos/farmacologia , Quimiometria , Computadores , Farmacorresistência Fúngica , Fluconazol/farmacologia , Humanos , Concentração de Íons de Hidrogênio , Testes de Sensibilidade MicrobianaRESUMO
AIMS: To evaluate the antimicrobial activity and to determine the pharmacodynamic characteristics of three 8-hydroxyquinoline derivatives (8-HQs) against Pythium insidiosum, the causative agent of pythiosis. METHODS AND RESULTS: Antimicrobial activity was tested by broth microdilution and MTT assays. The antimicrobial mode of action was investigated using sorbitol protection assay, ergosterol binding assay, and scanning electron microscopy. Clioquinol, PH151, and PH153 were active against all isolates, with MIC values ranging from 0.25 to 2 µg ml-1. They also showed a time- and dose-dependent antimicrobial effect, damaging the P. insidiosum cell wall. CONCLUSIONS: Together, these results reinforce the potential of 8-HQs for developing new drugs to treat pythiosis.
RESUMO
Often associated to the colonization by Candida spp. biofilm, the catheter-related infections are a serious health problem since the absence of a specific therapy. Hence, the main objective of this work was to evaluate the activity of 8-hydroxyquinoline and quinazoline derivatives on Candida spp. biofilms. A quinazoline derivative (PH100) and an 8-hydroxyquinoline derivative (PH157) were tested against nine strains of C. albicans, C. tropicalis and C. parapsilosis, and their biofilms in polystyrene microtitre plates and on polyurethane central venous catheter. The PH157 compound was incorporated into a film-forming system-type formulation and its capacity to inhibit biofilm formation on catheters was evaluated. The compounds were active against planktonic and sessile cells, as well as against the tested biofilms. PH157 compound performed better than the PH100 compound. The formulation containing PH157 presented results very similar to those of the compound in solution, which indicates that its activity was preserved. Both compounds showed activity against Candida spp. strains and their biofilm, with better PH157 activity. The formulation preserved the action of the PH157 compound, in addition, it facilitates its application on the catheter. The structural modifications that these compounds allow can generate compounds that are even more active, both against planktonic cells and biofilms.
Assuntos
Candida , Oxiquinolina , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Biofilmes , Testes de Sensibilidade Microbiana , QuinazolinasRESUMO
AIM: The purpose of this study was to evaluate the in vitro and in vivo efficiency of derivatives of 8-Hydroxyquinoline (8HQ) in controlling the fungus Ilyonectria liriodendri. METHODS AND RESULTS: The in vitro tests consisted of assessing its susceptibility to the minimal inhibitory concentration (MIC) and the inhibition of mycelial growth. While the in vivo tests consisted of applying and assessing the most effective products for the protection of wounds, in both preventive + curative and curative forms. The MIC values for PH 151 (6·25 µg ml-1 ) showed better results when compared to the fungicides tebuconazole (>50 µg ml-1 ) and mancozeb (12·5 µg ml-1 for strain 176 and 25 µg ml-1 for strain 1117). PH 151 significantly inhibited mycelial growth, while mancozeb did not differ from the control. In in vivo tests, PH 151 again demonstrated excellent results in vitro, especially when applied preventively. CONCLUSIONS: The derivative of 8HQ PH 151 was effective in controlling the fungus I. liriodendri in vitro and proved to be a promising option for protecting wounds. SIGNIFICANCE AND IMPACT OF THE STUDY: This study points to the prospect of an effective and safe preventive antifungal product, which would enable the use of pesticides in vine culture to be reduced.
Assuntos
Fungicidas Industriais/farmacologia , Hypocreales , Oxiquinolina , Doenças das Plantas , Vitis/microbiologia , Hypocreales/patogenicidade , Testes de Sensibilidade Microbiana , Oxiquinolina/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controleRESUMO
AIM: The purpose of this study was to evaluate the antifungal activity and toxicological parameters of 8-hydroxyquinoline derivatives PH151 and PH153 using alternative animal models, to understand their behaviour when subjected to in vivo experiments. METHODS AND RESULTS: We used Toll-deficient Drosophila melanogaster to test the protective effect of compounds against Candida albicans infection. Toxicological parameters were investigated in chicken and zebrafish embryos. PH151 and PH153 showed low toxicity and the treated flies with these compounds had a significantly higher survival rate than untreated flies after 7 days of infection. The compounds did not cause interruption of chicken embryogenesis. Zebrafish embryos exposed to compounds showed dose-dependent toxicity. CONCLUSIONS: The data supported the potential of PH151 and PH153 for the treatment of systemic candidiasis and demonstrated to be appropriate drug candidates for further studies using mammalian models. SIGNIFICANCE AND IMPACT OF THE STUDY: The increased incidence of Candida infections resistant to antifungals currently available requires acceleration of the discovery of new agents with properties of inhibiting this fungal pathogen. In this study, we have described the antifungal potential and toxicity of two 8-hydroxyquinoline derivatives using in vivo alternative models, and the results confirm their potential to be developed as new drug candidates.
Assuntos
Antifúngicos/uso terapêutico , Candidíase/tratamento farmacológico , Modelos Animais de Doenças , Oxiquinolina/uso terapêutico , Sulfonamidas/uso terapêutico , Animais , Antifúngicos/química , Candida albicans/efeitos dos fármacos , Candidíase/microbiologia , Embrião de Galinha , Drosophila melanogaster , Oxiquinolina/química , Sulfonamidas/química , Peixe-ZebraRESUMO
AIM: The purpose of this study was to uncover insights into the mechanism of action of the 8-hydroxyquinoline derivatives PH151 and PH153. In addition, with the future perspective of developing a topical drug for the treatment of candidiasis and dermatophytosis, the antifungal activity of a nanoemulsion formulation containing the most active compound (PH151) is also presented here. METHODS AND RESULTS: Sorbitol protection assay and scanning electron microscopy indicate that the 8-hydroxyquinoline derivatives act on the cell wall of Candida sp. and dermatophytes and they inhibit the pseudohyphae formation of C. albicans. These findings demonstrate a strong effect of these compounds on C. albicans morphogenesis, which can be considered a potential mode of action for this molecule. Besides, the nanoemulsion formulation MIC values ranged from 0·5 to 4 µg ml-1 demonstrating the significant antifungal activity when incorporated into a pharmaceutical formulation. CONCLUSIONS: Taken together, the results support the potential of these molecules as promising antifungal candidates for the treatment of candidiasis and dermatophytosis. SIGNIFICANCE AND IMPACT OF THE STUDY: There is an emerging need to fill the pipeline with new antifungal drugs due to the limitations presented by the currently used drugs. In this study, we have described a novel formulation with a 8-hydroxyquinoline-5-sulfonamide derivative which has presented a great potency in providing a finished product. Furthermore, the derivative has shown a selective mechanism of action confirming its potential to be developed into a new drug candidate.
Assuntos
Antifúngicos/farmacologia , Arthrodermataceae/efeitos dos fármacos , Dermatomicoses/microbiologia , Oxiquinolina/farmacologia , Sulfonamidas/farmacologia , Antifúngicos/química , Arthrodermataceae/crescimento & desenvolvimento , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Candidíase/tratamento farmacológico , Candidíase/microbiologia , Parede Celular/efeitos dos fármacos , Dermatomicoses/tratamento farmacológico , Hifas/efeitos dos fármacos , Hifas/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Oxiquinolina/química , Sulfonamidas/químicaRESUMO
Undeniably, new antifungal treatments are necessary against pathogenic fungi. Fungal infections have significantly increased in recent decades, being highlighted as important causes of morbidity and mortality, particularly in immunocompromised patients. Five main antifungal classes are used: (i) azoles, (ii) echinocandins, (iii) polyenes, (iv) allylamines and (v) pyrimidine analogues. Moreover, the treatment of mycoses has several limitations, such as undesirable side effects, narrow activity spectrum, a small number of targets and fungal resistance, which are still of major concern in clinical practice. The discovery of new antifungals is mostly achieved by the screening of natural or synthetic/semisynthetic chemical compounds. The most recent discoveries in drug resistance mechanism and their avoidance were explored in a review, focusing on different antifungal targets, as well as new agents or strategies, such as combination therapy, that could improve antifungal therapy. SIGNIFICANCE AND IMPACT OF THE STUDY: The failure to respond to antifungal therapy is complex and is associated with microbiological resistance and increased expression of virulence in fungal pathogens. Thus, this review offers an overview of current challenges in the treatment of fungal infections associated with increased antifungal drug resistance and the formation of biofilms in these opportunistic pathogens. Furthermore, the most recent and potential strategies to combat fungal pathogens are explored here, focusing on new agents as well as innovative approaches, such as combination therapy between antifungal drugs or with natural compounds.
Assuntos
Antifúngicos/farmacologia , Farmacorresistência Fúngica , Fungos/efeitos dos fármacos , Micoses/microbiologia , Animais , Descoberta de Drogas , Fungos/genética , Fungos/fisiologia , Humanos , Micoses/tratamento farmacológicoRESUMO
AIMS: Evaluate the in vitro effect of imidazolium salts (IMS) on the conidia germination and mycelial growth of Fusarium graminearum and their in vivo efficacy for suppressing the symptoms of the disease and infection of kernels in wheat plants. METHODS AND RESULTS: The minimum inhibitory concentrations (MIC) of three IMS (C16 MImCl, C16 MImMeS and C16 MImNTf2 ) were determined for four F. graminearum isolates using serial broth dilution method. The MICs found for all IMS were either 3·12 or 6·25 µg ml(-1) across the isolates, with the former as the most frequent. In the mycelial growth assay on potato dextrose agar media, only the C16 MImCl among the IMS reduced 50% of mycelial growth of one isolate at an estimated concentration of 0·32 mg ml(-1) . The time-kill curves showed a strong fungicidal effect starting 1 h after incubation at a concentration of 12·5 µg ml(-1) , representing a fourfold increase in the most frequent MIC. The C16 MImCl sprayed onto the spikes of potted wheat plants during the flowering stage reduced disease intensity at levels comparable to the commercial fungicide when applied preventatively (1 h prior to fungal inoculation), rather than curatively, and at the higher dosage (2 mg ml(-1) ) rather than lower dosage (0·5 mg ml(-1) ). CONCLUSIONS: C16 MImCl proved to be a potent inhibitor of F. graminearum growth and provided good levels of control of the disease at levels comparable to a commercial fungicide, in wheat plants treated prior to fungal infection during flowering stages. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests the potential of using IMS as alternative to the hazardous standard fungicides in the management of Fusarium head blight of wheat.
Assuntos
Fungicidas Industriais/farmacologia , Fusarium/efeitos dos fármacos , Imidazóis/farmacologia , Doenças das Plantas/microbiologia , Triticum/microbiologia , Fusarium/crescimento & desenvolvimento , Fusarium/fisiologia , Testes de Sensibilidade Microbiana , Doenças das Plantas/prevenção & controle , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/fisiologiaRESUMO
AIMS: This study demonstrates the development of a mouthwash formulation containing the imidazolium salt (IMS) 1-n-hexadecyl-3-methylimidazolium chloride (C16 MImCl), considering its stability and efficacy against Candida sp. Biofilm formation. METHODS AND RESULTS: A variety of in vitro test methods were applied, assessing contaminated acrylic resin strip specimens before and after applying the mouthwash formulations. The formulation using C16 MImCl presented a similar antibiofilm activity to cetylpyridinium chloride one and a commercial mouthwash, but at a 10 times lower concentration. Scanning electron microscopy imaging demonstrated that the selected mouthwash preparation fully destroys the biofilm cells, while with the hypoallergenicity test no irritant effect was observed in ex vivo model. CONCLUSIONS: The results presented herein indicate a high potential for imidazolium salts application as mouthwash agents that can eliminate Candida biofilm growth at very low concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates a new and effective antibiofilm formulation containing the IMS C16 MImCl. These findings suggest the IMS' use as mouthwash formulations active ingredient against Candida biofilms on oral surfaces, as it outperforms the often used cetylpyridinium chloride at a 10 times lower concentration.
Assuntos
Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Imidazóis/farmacologia , Antissépticos Bucais/farmacologia , Biofilmes/efeitos dos fármacos , Candida/fisiologiaRESUMO
AIMS: To evaluate the ability of Candida parapsilosis and Candida glabrata to develop phenotypic resistance to a benzophenone enriched fraction obtained from Brazilian red propolis (BZP-BRP) as compared to fluconazole (FLC). To investigate possible synergy between BZP-BRP and FLC and anidulafungin (AND). METHODS AND RESULTS: To analyse the development of resistance, isolates susceptible to these antifungals were cultured in increasing concentrations of FLC and BZP-BRP. The increase in FLC minimum inhibitory concentration for all isolates was evident and the majority developed resistance, whereas none isolated became less susceptible to BZP-BRP. Synergism was investigated by checkerboard method. BZP-BRP demonstrated synergy with FLC and indifference with AND for most isolates. CONCLUSIONS: In conclusion, the synergism observed with FLC suggests that BZP-BRP could be a possible therapeutic strategy for the treatment of infections related to FLC-resistant Candida sp. SIGNIFICANCE AND IMPACT OF THE STUDY: The indiscriminate use of antifungals results in the emergence of drug-resistant strains among previously susceptible populations. BZP-BRP can become an alternative for the treatment of persistent infections caused by Candida sp.
Assuntos
Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Extratos Vegetais/farmacologia , Própole/química , Brasil , Candida/classificação , Candida/genética , Candida/metabolismo , Farmacorresistência Fúngica , Sinergismo Farmacológico , Fluconazol/farmacologia , Testes de Sensibilidade MicrobianaRESUMO
AIMS: To investigate the antidermatophytic action of a complementary set imidazolium salts (IMS), determining structure-activity relationships and characterizing the IMS toxicological profiles. METHODS AND RESULTS: The susceptibility evaluation of 45 dermatophytic clinical isolates, treated in vitro with eleven different IMS (ionic compounds) and commercial antifungals (nonionic compounds), was performed by broth microdilution, following the standard norm of CLSI M38-A2. All dermatophytes were inhibited by IMS, where the lowest minimum inhibitory concentration (MIC) values were observed for salts with n-hexadecyl segment in the cation side chain, containing either the chloride or methanesulfonate anion. 1-n-Hexadecyl-3-methylimidazolium chloride (C16 MImCl) and 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16 MImMeS) acted as fungicides, even in extremely low concentrations, wherein C16 MImMeS exerted this effect on 100% of the tested dermatophytes. Some of these IMS provoked evident alterations on the fungi cell morphology, causing a total cell damage of ≥ 70%. Importantly, none of the screened IMS were cytotoxic, mutagenic or genotoxic to human leucocyte cells. CONCLUSIONS: This report demonstrates for the first time the strong antifungal potential of IMS against multidrug-resistant dermatophytes, without presenting toxicity to human leucocyte cells at MIC. SIGNIFICANCE AND IMPACT OF THE STUDY: The expressive antifungal activity of IMS, combined with the in vitro nontoxicity, makes them promising compounds for the safe and effective treatment of dermatophytoses, mainly when this skin mycosis is unresponsive to conventional drugs.
Assuntos
Antifúngicos/farmacologia , Arthrodermataceae/efeitos dos fármacos , Dermatomicoses/microbiologia , Farmacorresistência Fúngica Múltipla , Imidazóis/farmacologia , Arthrodermataceae/crescimento & desenvolvimento , Dermatomicoses/tratamento farmacológico , Humanos , Testes de Sensibilidade MicrobianaRESUMO
UNLABELLED: Although the use of catheters in critically ill patients is mostly inevitable, this invasive procedure comes together with several health risks. Within this context, the contamination with Candida tropicalis is a primary concern as this highly prevalent pathogenic yeast can develop an extensive polymeric matrix that hinders the drugs' penetration and its effective treatment. This study addresses the potential for the 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16 MImMeS) and chloride (C16 MImCl) salts for eliminating the viable cells of biofilms of Candida tropicalis, compared to the performance of chlorhexidine (CHX) and fluconazole (FLZ). The minimum concentration required of C16 MImMeS, C16 MImCl, CHX and FLZ for elimination of the biofilm's viable cells (MBEC) was evaluated through microtitre plate biofilm exposure with different concentrations of these substances. These concentrations were determined at 80% of effective activity against the biofilm's viable cells by using the MTT reduction assay. C16 MImMeS and C16 MImCl were able to eliminate the viable cells at much lower concentrations (15·6 and 0·45 µg ml(-1) respectively) than CHX (1250 µg ml(-1) ) and FLZ (resistance of the viable cells). This demonstrates the high potential of these substances for nosocomial infections control. SIGNIFICANCE AND IMPACT OF THE STUDY: The 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16 MImMeS) and chloride (C16 MImCl) salts are extremely effective in eliminating the viable cells of Candida tropicalis biofilms, which allows the use of much lower concentrations than with the antimicrobial of choice (chlorhexidine) in hospital practices. These findings indicate these imidazolium salts as high-potential candidates for asepsis of medical environments and materials, including implants.
Assuntos
Biofilmes/efeitos dos fármacos , Candida tropicalis/efeitos dos fármacos , Infecções Relacionadas a Cateter/prevenção & controle , Imidazóis/farmacologia , Anti-Infecciosos/farmacologia , Antifúngicos/farmacologia , Infecções Relacionadas a Cateter/microbiologia , Cateteres Venosos Centrais/microbiologia , Clorexidina/farmacologia , Infecção Hospitalar/prevenção & controle , Fluconazol/farmacologia , Humanos , Mesilatos/farmacologia , Testes de Sensibilidade Microbiana , Sais/farmacologiaRESUMO
UNLABELLED: The in vitro activity of the imidazolium salt C16 MImCl against planktonic and biofilm cells of multidrug-resistant isolates of Candida tropicalis was evaluated, both in solution and applied on a commercial catheter surface. This was determined by inhibition and susceptibility assays of biofilm and planktonic cells. In both cases, C16 MImCl prevented in vitro biofilm formation of C. tropicalis strains, including multidrug-resistant ones. Outstanding performances were observed, even at extremely low concentrations. Furthermore, this is the first report of the antifungal lock property of C16 MImCl, using a tracheal catheter as the test specimen to mimic a clinical in vivo condition. As such, C16 MImCl has been identified as a promising antimicotic pharmaceutical candidate for the treatment of candidiasis infections. SIGNIFICANCE AND IMPACT OF THE STUDY: The imidazolium salt 1-n-hexadecyl-3-methylimidazolium chloride (C16 MImCl) strongly prevents, in concentrations as low as 0·028 µg ml(-1) , the biofilm formation of multidrug-resistant Candida tropicalis isolates, either in solution or applied on the surface of commercial catheters. This presents an effective antimicotic candidate and alternative for invasive clinical procedure toolset asepsis.
Assuntos
Antifúngicos/farmacologia , Candida tropicalis/efeitos dos fármacos , Imidazóis/farmacologia , Biofilmes/efeitos dos fármacos , Candida tropicalis/isolamento & purificação , Candida tropicalis/fisiologia , Candidíase/microbiologia , Farmacorresistência Fúngica Múltipla , Humanos , Testes de Sensibilidade Microbiana , Plâncton/efeitos dos fármacosRESUMO
The aim of this work was to develop solid lipid nanoparticles (SLN) containing copaiba oil with and without allantoin (NCOA, NCO, respectively) and to evaluate their antifungal activity. Nanoparticle suspensions were prepared using a high homogenisation technique and characterised by dynamic light scattering, laser diffraction, nanoparticle tracking analysis, multiple light scattering analysis, high-pressure liquid chromatography, pH and rheology. The antifungal activities of the formulations were tested in vitro against the emergent yeasts Candida krusei and Candida parapsilosis, and the fungal pathogens of human skin Trichophyton rubrum and Microsporum canis. The dynamic light scattering analysis showed z-average diameters (intensity) between 118.63 ± 8.89 nm for the nanoparticles with both copaiba oil and allantoin and 126.06 ± 9.84nm for the nanoparticles with just copaiba oil. The D[4,3] determined by laser diffraction showed similar results of 123 ± 1.73 nm for the nanoparticles with copaiba oil and allantoin and 130 ± 3.6 nm for the nanoparticles with copaiba oil alone. Nanoparticle tracking analysis demonstrated that both suspensions had monomodal profiles and consequently, the nanoparticle populations were homogeneous. This analysis also corroborated the results of dynamic light scattering and laser diffraction, exhibiting a smaller mean diameter for the nanoparticles with copaiba oil and allantoin (143 nm) than for the nanoparticles with copaiba oil (204 nm). The physicochemical properties indicated that the dispersions were stable overtime. Rheology evidenced Newtonian behaviour for both suspensions. Antifungal susceptibility showed a MIC90 of 125 µg/mL (nanoparticles with copaiba oil) and 7.8 µg/mL (nanoparticles with copaiba oil and allantoin) against C. parapsilosis. The nanoparticles with copaiba oil and the nanoparticles with copaiba oil and allantoin presented a MIC90 of 500 µg/mL and 250 µg/mL, respectively, against C. krusei. The MIC90 values were 500 µg/mL (nanoparticles with copaiba oil) and 1.95 µg/mL (nanoparticles with copaiba oil and allantoin) against T. rubrum. Against M. canis, the nanoparticles with copaiba oil and allantoin had a MIC9 of 1.95 µg/mL. In conclusion, nanoencapsulation improved the antifungal activity of copaiba oil, which was enhanced by the presence of allantoin. The MICs obtained are comparable to those of commercial products and can represent promising therapeutics for cutaneous infections caused by yeasts and dermatophytes.
Assuntos
Alantoína/química , Alantoína/farmacologia , Antifúngicos/química , Antifúngicos/farmacologia , Composição de Medicamentos/métodos , Fabaceae/química , Lipídeos/química , Nanopartículas/química , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Alantoína/administração & dosagem , Antifúngicos/administração & dosagem , Química Farmacêutica , Fungos/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Tamanho da Partícula , Óleos de Plantas/administração & dosagem , ReologiaRESUMO
In this study, we investigate the ability of Pythium insidiosum to form biofilms across various substrates and the antibiofilm efficacy of 8-hydroxyquinoline derivatives (8-HQs). Biofilms of P. insidiosum were cultured on polystyrene plates, contact lenses, and horsehair. We provide the first evidence of P. insidiosum's biofilm-forming capability, thus considerably expanding our understanding of its transmission and pathogenesis. Our results demonstrate that 8-HQs effectively inhibit biofilm formation and eradicate pre-existing biofilms, underscoring their potential as a novel treatment strategy for pythiosis, a disease currently lacking a gold-standard treatment. This finding has particular relevance for ocular pythiosis associated with contact lens usage and potential infection sources in animals. Our results contribute to the scientific knowledge base and directly impact innovative therapeutic interventions' development.
Assuntos
Pitiose , Pythium , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Pitiose/tratamento farmacológico , Pitiose/microbiologiaRESUMO
INTRODUCTION: Itraconazole is the first-choice option to treat human and animal sporotrichosis. However, the emergence of itraconazole-resistant strains has encouraged research on new active antifungals. Among them, the essential oil of rosemary (Rosmarinus officinalis Linn., Lamiaceae) has shown antifungal activity in vitro. OBJECTIVE: Assessing, for the first time, the effectiveness of rosemary essential oil in vivo in experimental cutaneous sporotrichosis, as well as its chemical composition and action mode. METHODS: Itraconazole-resistant Sporothrix brasiliensis was inoculated in the left foot pad of 30 Wistar rats, which were randomized (n=10) for treatment with saline solution (control, CONT), itraconazole (ITRA, 10 mg/kg) and rosemary oil (ROSM, 250 mg/kg) for 30 days at an oral dose of 1 mL, daily. Clinical evolution, histopathology and fungal burden were investigated. GC-MS was used for chemical analysis; sorbitol protection and ergosterol effect were used to evaluate the action mechanism of rosemary oil. RESULTS: ROSM was the only group evolving to skin lesion remission, lack of edema and exudate, and mild-to-absent yeast cells. Rosemary oil delayed fungal spreading and protected systemic organs, mainly liver and spleen. The ROSM group presented lower fungal load than that observed for the CONT and ITRA groups (p<0.05). Antifungal action took place at complexation level after ergosterol application. Most compounds were 1,8-cineole/eucalyptol (47.91%), camphor (17.92%), and α-pinene (11.52%). CONCLUSIONS: These findings have evidenced that rosemary oil is a promising antifungal to treat sporotrichosis, since it protects systemic organs from fungal spread.
Assuntos
Óleos Voláteis , Rosmarinus , Animais , Ratos , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Itraconazol/farmacologia , Óleos Voláteis/farmacologia , Ratos Wistar , SporothrixRESUMO
Onychomycosis are fungal nail infections comprising of about 50% of onychopathies and are commonly caused by dermatophytes. The treatment of this dermatomycosis requires a long period of time and is associated with high rates of recurrence. In view of the need to evaluate the antifungal performance of promising preclinical compounds, we developed, in this study, a practical and accessibleex vivo model for establishing a Trichophyton rubrum onychomycosis framework using porcine hooves. This model has as its main advantage the similar structural and three-dimensional characteristics that the porcine hooves have with the human nail. The proposed model allowed to evaluate the antifungal activity of a new antifungal compound and a reference drug (terbinafine), both already incorporated into a nail lacquer for topical use. Treatments with compound 3-selenocyanate-indole (Se4a) and with terbinafine incorporated into this nail lacquer completely inhibited fungal growth, corresponding to the profile of in vitro activity observed against T. rubrum. This study concludes that the ex vivo porcine hoof model is an effective alternative method for preclinical screening of drugs or new topical compounds developed to combat onychomycosis. Further studies are needed to compare the permeability of porcine hooves with human nails permeability.
Assuntos
Antifúngicos/administração & dosagem , Avaliação Pré-Clínica de Medicamentos/métodos , Casco e Garras/patologia , Onicomicose/tratamento farmacológico , Técnicas de Cultura de Órgãos , Suínos , Administração Tópica , Animais , Antifúngicos/farmacologia , Cianatos/química , Casco e Garras/efeitos dos fármacos , Humanos , Laca , Testes de Sensibilidade Microbiana/métodos , Modelos Biológicos , Onicomicose/patologia , Permeabilidade/efeitos dos fármacos , Compostos de Selênio/química , Terbinafina/administração & dosagem , Terbinafina/farmacologia , Trichophyton/efeitos dos fármacos , Trichophyton/crescimento & desenvolvimentoRESUMO
BACKGROUND: Intensive prophylactic use of antifungals leads to the increase of drug resistance and the need for new and more effective treatments are real. Plants from Leguminosae family are rich in flavonoids, for which numerous biological activities have been described, including antifungal effects. PURPOSE: To screen methanolic extracts from Leguminosae species looking for alternative sources for antifungal agents (anti-dermatophyte and anti-Candida) and their innocuity. METHODS: Antifungal activity was evaluated using the strains Candida albicans, C. krusei, C. glabrata, C. tropicalis, C. parapsilosis, Epidermophyton floccosum, Trichophyton mentagrophytes, T. rubrum and, Microsporum gypseum in the broth microdilution method. Later, the minimum inhibitory concentration (MIC) for Mimosa pigra, Eriosema heterophyllum, and Chamaecrista nictitans was determined. The most promising extract was fractionated and cytotoxicity and genotoxicity of the most active fraction were also assayed. RESULTS: Fungicide and/or fungistatic activity against dermatophyte strains were presented by 60% of the methanolic extracts assayed. M. pigra, E. heterophyllum, and C. nictitans methanolic extracts could inhibit dermatophyte strains at concentrations ranging from 1.9 to 1000µg/mL. M. pigra showed the lowest MIC values for a dichloromethane fraction (1.9µg/mL) without DNA damage at 10 and 50µg/mL and 100% of cell viability of human leukocytes. CONCLUSION: Our results indicate that methanolic extracts from Leguminosae plants are potential sources of antifungal compounds, mainly the extract and fractions from M. pigra. The dichloromethane fraction from M. pigra did not showed in vitro toxicity according to the applied assays.
Assuntos
Antifúngicos/farmacologia , Arthrodermataceae/efeitos dos fármacos , Fabaceae/química , Mimosa/química , Extratos Vegetais/farmacologia , Brasil , Candida/efeitos dos fármacos , Epidermophyton/efeitos dos fármacos , Humanos , Masculino , Testes de Sensibilidade Microbiana , Microsporum/efeitos dos fármacos , Testes de Toxicidade , Trichophyton/efeitos dos fármacosRESUMO
Este trabalho teve como objetivo, avaliar a prevalência no diagnóstico de dermatófitos durante o período de janeiro de 2007 à junho de 2008 no setor de micologia em hospital particular de médio porte, na cidade de Chapecó, oeste do estado de Santa Catarina. Foram coletadas 111 amostras, das quais 66 (59%) apresentaram positividade pelo exame direto e cultivo da amostra biológica. Trichophyton mentagrophytes foi o fungo isolado com maior freqüência (52%), seguido pelo dermatófito T. rubrum (17%), em contrapartida dos dados literários no sul do Brasil, que preconizam T. rubrum, seguido de Microsporum canis e do T. mentagrophytes como agentes mais comumente isolados. Considerando os sítios anatômicos analisados neste trabalho, a ocorrência foi observada em 47% em amostras de unha, 43% de pele, 7% outros e 3% mistos (pele/unha). Esse estudo evidencia a importância da recorrente análise do perfil epidemiológico dos dermatófitos nas diferentes regiões do Brasil, possibilitando uma correta conduta epidemiológica de prevenção, baseada na freqüência regional das espécies causadoras das dermatomicoses.
The aim of this study was to assess the prevalence of dermatophytes diagnosed, from January 2007 to June 2008, at the clinical mycology section of a private medium-sized hospital, in Chapecó city (Santa Catarina state, Brazil). Out of the 111 samples collected, 66 (59%) gave positive results in the direct examination and culture of the biological sample. Trichophyton mentagrophytes was the most frequently isolated species (52%), followed by T. rubrum (17%), contradicting the published data on southern Brazil, which identify T. rubrum as the commonest agent in clinical specimens, followed by Microsporum canis and T. mentagrophytes. Regarding the anatomical sites from which the isolates were taken, 47% were found in nail samples, 43% in skin, 7% at other sites and 3% mixed (skin/nail). This study highlights the importance of regularly analyzing the epidemiological profile of dermatophytes in each region of Brazil, to enable epidemiological prevention to be carried out effectively, based on the frequencies of the species causing dermatomycoses in the regional population.