Antifungal activity of azoles, allylamines, and 8-hidroxiquinolines, alone and in combination, against Malassezia pachydermatis in vitro and in vivo.

Malassezia pachydermatis is often reported as the causative agent of dermatitis in dogs. This study aims to evaluate the in vitro and in vivo antifungal activity of azoles and terbinafine (TRB), alone and in combination with the 8-hydroxyquinoline derivatives (8-HQs) clioquinol (CQL), 8-hydroxyquinoline-5-(n-4-chlorophenyl)sulfonamide (PH151), and 8-hydroxyquinoline-5-(n-4-methoxyphenyl)sulfonamide (PH153), against 16 M. pachydermatis isolates. Susceptibility to the drugs was evaluated by in vitro broth microdilution and time-kill assays. The Toll-deficient Drosophila melanogaster fly model was used to assess the efficacy of drugs in vivo. In vitro tests showed that ketoconazole (KTZ) was the most active drug, followed by TRB and CQL. The combinations itraconazole (ITZ)+CQL and ITZ+PH151 resulted in the highest percentages of synergism and none of the combinations resulted in antagonism. TRB showed the highest survival rates after seven days of treatment of the flies, followed by CQL and ITZ, whereas the evaluation of fungal burden of dead flies showed a greater fungicidal effect of azoles when compared to the other drugs. Here we showed for the first time that CQL is effective against M. pachydermatis and potentially interesting for the treatment of malasseziosis.

Infrared spectroscopy combined with chemometrics in transflectance mode: An alternative approach in the photodiagnosis of COVID-19 using saliva.

The Coronavirus Disease 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has required the search for sensitive, rapid, specific, and lower-cost diagnostic methods to meet the high demand. The gold standard method of laboratory diagnosis is real-time reverse transcription polymerase chain reaction (RT-PCR). However, this method is costly and results can take time. In the literature, several studies have already described the potential of Fourier transform infrared spectroscopy (FTIR) as a tool in the biomedical field, including the diagnosis of viral infections, while being fast and inexpensive. In view of this, the objective of this study was to develop an FTIR model for the diagnosis of COVID-19. For this analysis, all private clients who had performed a face-to-face collection at the Univates Clinical Analysis Laboratory (LAC Univates) within a period of six months were invited to participate. Data from clients who agreed to participate in the study were collected, as well as nasopharyngeal secretions and a saliva sample. For the development of models, the RT-PCR result of nasopharyngeal secretions was used as a reference method. Absorptions with high discrimination (p < 0.001) between GI (28 patients, RT-PCR test positive to SARS-CoV-2 virus) and GII (173 patients who did not have the virus detected in the test) were most relevant at 3512 cm-1, 3385 cm-1 and 1321 cm-1 after 2nd derivative data transformation. To carry out the diagnostic modeling, chemometrics via FTIR and Discriminant Analysis of Orthogonal Partial Least Squares (OPLS-DA) by salivary transflectance mode with one latent variable and one orthogonal signal correction component were used. The model generated predictions with 100 % sensitivity, specificity and accuracy. With the proposed model, in a single application of an individual's saliva in the FTIR equipment, results related to the detection of SARS-CoV-2 can be obtained in a few minutes of spectral evaluation.

Prediction of melanin content of Fonsecaea pedrosoi using Fourier transform infrared spectroscopy (FTIR) and chemometrics.

Fungal melanin contributes to the survival and virulence of pathogenic fungi, such as Fonsecaea pedrosoi, which is responsible for causing chromoblastomycosis. The objective of this study was to employ Fourier transform infrared spectroscopy (FTIR) to predict the melanin content of F. pedrosoi. The melanin content, in percentage, was previously determined using gravimetry for twenty-six clinical isolates. Quintuplicate spectra of each isolate were obtained using attenuated total reflection (ATR) within the range of 4000 to 650 cm-1. To predict the melanin content, modeling was performed using partial least squares regression (PLS) in the region 1800 - 750 cm-1. Two models were tested: PLS and successive projections algorithms for interval selection in partial least squares (iSPA-PLS). The best modeling results were achieved using iSPA-PLS with one factor. The calibration set exhibited a determination coefficient (R2) of 0.9745 and a root mean square error of cross-validation (RMSECV) of 0.0977. In the prediction set, the R2 value was 0.9711, and the root mean square error of prediction (RMSEP) was 0.0999. Modeling with FTIR and multivariate calibration provides a valuable means of predicting fungal melanin content, which is simpler and more robust, thereby contributing to the advancement of this field of study.

Chemical characterization and antimicrobial activity of Baccharis vulneraria Baker essential oil against strains of microorganisms that cause cutaneous infections.

Baccharis vulneraria Baker is used popularly for the treatment of skin infections. So, this study aimed investigate the antimicrobial activity and chemical characterization of the essential oil (EO) against microorganisms that cause cutaneous infections. The EO was analyzed by GC-MS. The antimicrobial test was performed using the serial microdilution method, and the antimicrobial activity was determined by the minimum inhibitory concentration against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Trichophyton interdigitale, Trichophyton rubrum, Fusarium solani and Fusarium oxysporum within the 32-0.0625 mg/mL concentration range. Were identified 31 EO compounds. Its major compounds are bicyclogermacrene, trans-cadin-1,4-diene, ß-caryophyllene, and germacrene A. EO showed antifungal action against T. rubrum and T. interdigitale (2 and 4 mg/mL MIC, respectively). The growth of C. albicans, at 4 mg/mL, decreased by 50% compared to control. The oil had no significant potential for other microorganisms at the selected concentrations.

High MICs for antifungal agents in yeasts from an anthropized lagoon in South America.

The lagoons are fragile ecosystems used by several species as a refuge and breeding area, and it is also a place where certain communities practice fishing activity. With increasing urbanization around this ecosystem, pesticides used in agriculture and untreated urban wastewater are drained into the river basin, resulting in the dispersion of organic matter and antifungals used by the population and farmers. These may favor the selection of resistant pathogens directly into the environment, a concern since several fungi have emerged as pathogens in the last decades. In this study, we investigated the presence in an impacted lagoon by potentially resistant yeasts to antifungal agents. We evaluated their capacity for producing extracellular enzymes that could act as virulence factors. Water samples from the Tramandaí lagoon were analyzed for the presence of pesticides using the SPE-LC-ESI-MS/MS. Tricyclazole, carbendazim, azoxystrobin, thiabendazole, and tebuconazole were found. Twenty-eight yeast species were isolated, including the multidrug-resistant Candida haemulonii, and species with high minimal inhibitory concentrations (MICs) for clinical antifungal agents. Around 93% of the isolates had MIC values above the resistance breakpoints established for Candida species for at least two antifungal agents. And 27% had high MICs values for fluconazole, terbinafine, amphotericin B, and caspofungin. Tebuconazole MICs values were highly associated with MICs for fluconazole, terbinafine, and amphotericin B, and significant correlations between high MICs for antifungal agents and enzyme production were found. The results indicated that the lagoon is a reservoir of resistance genes and a potential source for fungal infection, highlighting the importance of the One Health approach and the integrated vision of the ecosystem when managing these environments.

Molecular identification and antifungal susceptibility of 75 clinical isolates of Trichophyton spp. from southern Brazil.

BACKGROUND: Dermatophytoses affect more than 20% of the world's population and is caused by filamentous fungi, mainly of the genus Trichophyton. The species identification through microscopic direct examination and culture methods is challenging, with molecular presenting high sensitivity and specificity. Although there are several therapy options for dermatophyte infections, treatment failures and antifungal resistance are growing concerns. OBJECTIVE: This study aimed to identify clinical isolates of Trichophyton spp. from southern Brazil using molecular methods and determine their in-vitro antifungal susceptibility. MATERIAL AND METHODS: Seventy-five isolates were identified through sequencing of the ITS region. The exposure to seven antifungals drugs was performed according to protocol M28-A2 of the Clinical and Laboratory Standards Institute (CLSI). RESULTS: Sixty-one isolates (81%) were identified as T. interdigitale, which differs from the epidemiological data present in the literature. Thirteen isolates were identified as T. rubrum and one as T. tonsurans. Terbinafine was the most effective antifungal, followed by itraconazole and voriconazole, which is in accordance with the results reported in previous studies. CONCLUSIONS: The use of molecular methods to identify Trichophyton spp. clinical isolates and the performance of susceptibility tests are relevant to epidemiological data, identification of the emergence of antifungal resistance, and to help to translate the in-vitro antifungal susceptibility results into clinical practice.

Evaluation of the activity of filamentous fungi isolated from soils of the Pampa biome applied in the biological control of Tetranychus urticae (Acari: Tetranychidae) and Polyphagotarsonemus latus (Acari: Tarsonemidae).

Tetranychus urticae Koch and Polyphagotarsonemus latus Banks are mite species considered capable of attaining pest levels, damaging a range of agricultural crops. The Pampa biome is characterized by the high biodiversity it houses, particularly microbial diversity, which highlights its potential for developing microorganisms that can provide biological control of arthropods. The aim of this study was to evaluate the activity of four fungal isolates from the soil of the Pampa biome in the biological control of T. urticae (females and eggs) and P. latus (females). Experiments consisted of isolating and identifying fungal isolates for spore quantification and aspersion at 108, 106, and 104 spores/mL concentrations in arenas containing T. urticae females and eggs, and P. latus females, separately. Results indicated that only three isolates (Aspergillus brunneoviolaceus, Clonostachys chloroleuca, and Penicillium adametzii) showed high control of T. urticae females, yet they did not exhibit any control of T. urticae eggs and P. latus females. Therefore, the present study confirms the viability of some of these fungi as biological control agents of mites, which implies the importance of new prospects with other fungal species, considering the richness of resources in the Pampa biome, or even the need to test higher concentrations and other variables using the microorganisms of the present study.

Degradation of micropollutant cephalexin by ultraviolet (UV) and assessment of residual antimicrobial activity of transformation products.

Cephalexin (CEX) is an antibiotic commonly used to treat bacterial infections in humans and animals. However, it is also a micropollutant. Thus, this study evaluated the degradation of CEX using ultraviolet irradiation (UV-C) and analyzed the by-products as well as their residual antimicrobial activity. A reactor with a mercury vapor lamp was used for the degradation. Irradiated CEX solutions were collected over a period of 4 hours and analyzed using high-performance liquid chromatography coupled with mass spectrometry. For the residual antimicrobial activity the susceptibility test was performed using Staphylococcus aureus and Escherichia coli microorganisms by broth microdilution. It was found that CEX, after treatment, generated a metabolite with a mass of 150 m/z in 15 min. A four- and eightfold increase in the minimum inhibitory concentration of the drug against S. aureus and E. coli could be observed, respectively, after 20 min. Therefore, this treatment proved to be effective in the degradation of CEX, being able to degrade 81% of the initial molecule of the drug in 20 min. Furthermore, the antimicrobial activity of the CEX solution decreased as the irradiation time increased, indicating loss of antimicrobial function of the initial CEX molecule and the resulting by-products.

Effect of Melanin Biosynthesis Inhibition on the Antifungal Susceptibility of Chromoblastomycosis Agents.

Chromoblastomycosis (CBM) is a chronic subcutaneous infection caused by genera of melanized fungi: Fonsecaea, Cladophialophora, Phialophora, Exophiala, and Rhinocladiella. Melanin is a virulence factor known to influence antifungal susceptibility. A specific inhibitor of melanin biosynthesis is tricyclazole. The aim of this study was to evaluate the effect of melanin inhibition on antifungal susceptibility of chromoblastomycosis agents and describe the susceptibility profiles of some unusual CBM agents. Seventy-six clinical isolates, representing 13 species of the five main genera of CBM agents, were studied. The antifungal susceptibility testing was performed according to the M38-A2 protocol of CLSI (Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi, 3rd ed., CLSI Standard M38, 2017). In the melanin inhibition test, 16 mg/liter of tricyclazole was added to the medium used in the inoculum preparation and the susceptibility assay. CBM agents were less susceptible to amphotericin B than azoles and terbinafine. The unusual species showed similar susceptibility profiles to those of other species of the same genera. With tricyclazole exposure, MICs of terbinafine, posaconazole, and itraconazole for Fonsecaea spp. significantly decreased (P < 0.05). For Phialophora spp., this reduction was significant for posaconazole and itraconazole. For the other genera, there was a reduction in MICs of terbinafine and itraconazole; however, the statistical tests were not significant. Melanin inhibition can increase the antifungal susceptibility of most CBM agents to itraconazole and terbinafine, the main drugs used in the disease treatment. This increased susceptibility may open up new possibilities for therapy in refractory cases of CBM and/or cases caused by resistant fungal strains. Further studies are needed to confirm the same results in vivo.

Rapid classification of chromoblastomycosis agents genera by infrared spectroscopy and chemometrics supervised by sequencing of rDNA regions.

Chromoblastomycosis (CBM) is a skin and subcutaneous infection caused by species of seven fungal genera. Identification of CBM species is performed by DNA sequencing of one or more genes, which becomes a time-consuming work. Fourier Transform Infrared Spectroscopy (FTIR) has been used for the identification of other microorganisms, however, only one CBM genus was evaluated by FTIR analysis to date. Therefore, the study is aimed to differentiate the CBM agents for identification at genera level using FTIR supervised by Internal Transcribed Spacer (ITS) rDNA region. Seventy-seven isolates of the main five CBM genera were prepared for Attenuated Total Reflection FTIR (ATR-FTIR) with a new methodology using slices of dry fungus in glass fixing-modeling proposed in this study. The algorithm Hierarchical Cluster Analysis (HCA) was used to analyze the differences and similarities between species through the spectra. Orthogonal Partial Least Square Discriminant Analysis (OPLS-DA) allowed to correctly classify all samples of five CBM genera. The ATR-FTIR/OPLS-DA models highlighted important contributions of regions attributed to NH and OH stretching, amide I of proteins, polysaccharides bands and fingerprint region for the complete differentiation of the genera investigated. Thus, FTIR can be a fast and inexpensive alternative for identification of CBM agents.

Melanin and chromoblastomycosis agents: Characterization, functions, and relation with antifungals.

Melanins are a diverse group of dark pigments with similar properties. In fungi, the most studied is the dihydroxynaphtalene (DHN)-melanin, present in several species including all the chromoblastomycosis agents, a chronic, disabling, and recalcitrant subcutaneous mycosis. It is synthesized in a pathway known as the pentaketide pathway, which has the agrochemical tricyclazole as an inhibitor, widely used in in vitro studies because it does not prevent the growth of fungi. There are different methodologies for qualitative and quantitative analyses of DHN-melanin, which made it possible to discover its important structural and antioxidant functions, with melanin acting as a protective factor against the host's immune system. Also, it can interact with some of the main antifungals of medical interest, reducing its activity and the susceptibility of fungi to these agents. This review aims to discuss the aspects of DHN-melanin, focusing on chromoblastomycosis, bringing the main findings of the published scientific studies, and highlighting the need for further research to understand this important fungal pathogenicity and a virulence factor.

Prediction of itraconazole minimum inhibitory concentration for Fonsecaea pedrosoi using Fourier Transform Infrared Spectroscopy (FTIR) and chemometrics.

Fonsecaea pedrosoi is one of the main agents of chromoblastomycosis, a chronic subcutaneous mycosis. Itraconazole (ITC) is the most used antifungal in its treatment, however, in vitro antifungal susceptibility tests are important to define the best therapy. These tests are standardized by the Clinical and Laboratory Standards Institute (CLSI), but these protocols have limitations such as the high complexity, cost and time to conduct. An alternative to in vitro susceptibility test, which overcomes these limitations, is FTIR. This study determined the minimum inhibitory concentration (MIC) of itraconazole for F. pedrosoi, using FTIR and chemometrics. The susceptibility to ITC of 36 strains of F. pedrosoi was determined according to CLSI and with the addition of tricyclazole (TCZ), to inhibit 1,8-dihydroxynaphthalene (DHN)-melanin biosynthesis. Strains were grown in Sabouraud agar and prepared for Attenuated Total Reflection (ATR)/FTIR. Partial least squares (PLS) regression was performed using leave-one-out cross-validation (by steps of quintuplicates), then tested on an external validation set. A coefficient of determination (R²) higher than 0.99 was obtained for both the MIC-ITC and MIC-ITC+TCZ ATR/PLS models, confirming a high correlation of the reference values with the ones predicted using the FTIR spectra. This is the first study to propose the use of FTIR and chemometric analyses according to the M38-A2 CLSI protocol to predict ITC MICs of F. pedrosoi. Considering the limitations of the conventional methods to test in vitro susceptibility, this is a promising methodology to be used for other microorganisms and drugs.

Onychomycosis caused by Arthrinium arundinis in leprosy patient: Case report.

We report a case of a 56-year-old Brazilian woman, with relapsing lepromatous leprosy, and onychomycosis caused by a non-dermatophyte filamentous fungi. The pathogenic fungi was identified as Arthrinium arundinis and treated with chemical abrasion of the nail with 40% urea and application of terbinafine cream. Onychomycosis caused by Arthrinium species is rare, and this is the second reported case.

Population growth of the stored product pest Tyrophagus putrescentiae (Acari: Acaridae) on environmentally and medically important fungi.

The stored food mite Tyrophagus putrescentiae (Schrank) (Acari: Acaridae) has been associated with the presence of several fungal species. The aims of this work were to evaluate T. putrescentiae population growth associated to environmental and medically important fungal species to determine on which fungal species populations of T. putrescentiae performs best, and to evaluate their ability to disperse each fungal species. First, 24 fungal species were inoculated separately in Petri dishes containing Sabouraud agar medium. One week after inoculation, 50 mites were added to each plate. On the 28th evaluation day, mites and eggs were counted in each plate, and 50 mites randomly collected from each replicate were transferred to new plates containing only Sabouraud agar medium. Then, mites, eggs, and fungal population were evaluated in each plate on day 28 again. The highest population increases were on Trichophyton mentagrophytes, Alternaria sp., Microsporum gypseum, and Aspergillus chevalieri. With Fusarium guttiforme and the medically important fungi Microsporum canis, M. gypseum, T. mentagrophytes, and Sporothrix sp., mites were observed to feed on whole mycelium. Only eight fungal species were dispersed by T. putrescentiae to the new Petri dishes: Aspergillus clavatus, Candida tropicalis, Candida albicans, Fusarium guttiforme, Hyphopichia burtonii, Penicillium citrinum, Rhizophus azygosporus, and Trichophyton mentagrophytes. The best performance of T. putrescentiae was found feeding on F. guttiforme, P. citrinum, and T. mentagrophytes. In conclusion, T. putrescentiae successfully used fungi as a food source, and it proved to be an important tool for disseminating both environmental and medically important fungi.

In vitro susceptibility of chromoblastomycosis agents to antifungal drugs: A systematic review.

Chromoblastomycosis (CBM) is a chronic granulomatous mycosis caused by dematiaceous fungi that affects cutaneous and subcutaneous tissues. The standard antifungal drug for treatment is itraconazole, followed by terbinafine. However, cure rates vary from 15% to 80% when these drugs are used as monotherapy. A systematic review of the in vitro susceptibility of CBM agents to antifungal drugs, alone and in combination, was conducted using the Cochrane methodology. Forty-seven search terms were included in the PICOS method of searching electronic databases. The search resulted in 35 studies, of which 8 evaluated antifungal drugs in combination. Based on minimum inhibitory concentrations (MICs), posaconazole, terbinafine, itraconazole and voriconazole were, in descending order, the most effective antifungal drugs against CBM agents in vitro. In drug combination studies, only terbinafine-voriconazole and itraconazole-caspofungin showed 100% synergy for Fonsecaea pedrosoi, Exophiala jeanselmei and Phialophora verrucosa. However, none of the combinations studied showed antagonism.

Rosemary, Castor Oils, and Propolis Extract: Activity Against Candida Albicans and Alterations on Properties of Dental Acrylic Resins.

PURPOSE: To evaluate the in vitro activity of 8% rosemary, 2% castor oils, and 12% propolis glycolic extract against Candida albicans, as well as the physical changes of properties in colorless and pink acrylic resins after immersion in these liquids. MATERIALS AND METHODS: Colorimetric, roughness, and Knoop microhardness assays were evaluated in 25 specimens distributed into five groups (3 test groups and 2 control groups - distilled water and hypochlorite 1%), totaling five specimens per group for each acrylic resin (colorless and pink). The specimens were individually immersed for 30 minutes in 10 mL of these liquids, washed, and dried once a week. They were maintained in distilled water at 37°C between processes during all experiments. The analyses were performed before immersion and in the 4th and/or 12th month. In vitro, 18 acrylic resins were exposed to C. albicans and, after a process of 30 minutes in immersion in the five groups cited and oil vehicle control of vesicle (liquid Vaseline), the specimens were washed and incubated for 24 hours in 37°C. The growth was determined by colony counting. For comparisons between the groups in each trial and the disinfection test, paired Student's t-tests and ANOVA with post hoc Tukey were performed by the SPSS program, considering α = 0.05. RESULTS: None of the liquids altered the microhardness, but all the natural compounds and 1% sodium hypochlorite (control) altered color and roughness after the 12th month of immersion in these agents. In the colorless specimens, 8% rosemary oil caused a color change similar to water, and less color and roughness alterations when compared to 2% castor oil and 1% sodium hypochlorite, respectively. There was no growth of yeast colonies after immersion in rosemary oil, propolis glycolic extract, and 1% sodium hypochlorite. CONCLUSION: Eight percent rosemary oil has the potential to be used as an acrylic resin disinfectant.

Melanin: Quantification and protection against oxidative stress in chromoblastomycosis agents.

Chromoblastomycosis (CBM) is a chronic cutaneous and subcutaneous infection caused by melanized fungal species. We quantified the extractable melanin of 77 strains of CBM agents distributed within five genera. Moreover, resistance to oxidative stress was evaluated in strains exposed or not to the melanin inhibitor tricyclazole. The median percentage of melanin mass extracted from dry fungal mass varied from 0.69 (Rhinocladiella similis) to 3.81 (Phialophora americana). Inhibition of melanin synthesis decreased survival rates to hydrogen peroxide. Together, these data highlight the importance of melanin in CBM agents.

Drosophila melanogaster as a model for the study of Malassezia pachydermatis infections.

Malassezia pachydermatis is a yeast that is commonly found in the skin of most animals. Changes in the physical, chemical or immunological processes of the skin may render the host more susceptible to the yeast, which then may cause otitis, dermatitis or, less often, systemic infection. We tested the pathogenicity of M. pachydermatis in wild-type (WT) and Toll-deficient Drosophila melanogaster. Flies were inoculated in the thorax with a needle previously dipped in inoculum concentrations ranging from 103 and 107 yeast cells/mL. After infection, flies were housed at 29 °C and mortality was evaluated daily until day seven. WT flies were resistant to the infection, whereas Toll-deficient flies showed inoculum-dependent mortality rates. Fungal burden, assessed by histopathological analysis and by counting the number of colony-forming units of dead flies, corroborated the results. The D. melanogaster model is a promising minihost model for future large-scale studies of virulence mechanisms and antifungal drug activity in malasseziosis.

Sporotrichosis by Sporothrix schenckii senso stricto with itraconazole resistance and terbinafine sensitivity observed in vitro and in vivo: Case report.

We report a case of a patient with lymphocutaneous sporotrichosis in the right upper limb. The fungus was identified as Sporothrix schenckii senso stricto by calmodulin gene sequencing. The initial treatment was itraconazole (200 mg/day), but in vitro antifungal susceptibility demonstrated high resistant to this and another six antifungals, with exception to terbinafine. The lesions did not regress with itraconazole treatment. Thus, 500 mg/day of terbinafine was prescribed and clinical cure was obtained after four months.

Genetic diversity and antifungal susceptibility of Fusarium isolates in onychomycosis.

Fusarium species have emerged as an important human pathogen in skin disease, onychomycosis, keratitis and invasive disease. Onychomycosis caused by Fusarium spp. The infection has been increasingly described in the immunocompetent and immunosuppressed hosts. Considering onychomycosis is a difficult to treat infection, and little is known about the genetic variability and susceptibility pattern of Fusarium spp., further studies are necessary to understand the pathogenesis and better to define the appropriate antifungal treatment for this infection. Accordingly, the objective of this study was to describe the in vitro susceptibility to different antifungal agents and the genetic diversity of 35 Fusarium isolated from patients with onychomycosis. Fusarium spp. were isolated predominantly from female Caucasians, and the most frequent anatomical location was the nail of the hallux. Results revealed that 25 (71.4%) of isolates belonged to the Fusarium solani species complex, followed by 10 (28.5%) isolates from the Fusarium oxysporum species complex. Noteworthy, the authors report the first case of Neocosmospora rubicola isolated from a patient with onychomycosis. Amphotericin B was the most effective antifungal agent against the majority of isolates (60%, MIC ≤4 µg/mL), followed by voriconazole (34.2%, MIC ≤4 µg/mL). In general, Fusarium species presented MIC values >64 µg/mL for fluconazole, itraconazole and terbinafine. Accurate pathogen identification, characterisation and susceptibility testing provide a better understanding of pathogenesis of Fusarium in onychomycosis.