Abstract Cryptococcosis is an infectious fungal disease widely studied for its epidemiological importance in the context of public health, given the high morbidity and mortality associated with this invasive fungal infection. Many cases of the disease present clinical resistance and progress to death, even in the presence of antifungal therapy. The prolonged use of triazole drugs to maintain the treatment of cryptococcosis in AIDS patients, can lead to selective pressure from mutant strains, among other resistance mechanisms, justifying the poor clinical evolution of some cases. In this study, a narrative review of the literature on the occurrence of antifungal resistance in cryptococcosis agents was performed. Publications from 2010 to 2022 that address this topic were selected using Google Scholars and Scopus website. Data from the studies were analyzed for the values of minimum inhibitory concentration (MIC) of drugs used in the management of cryptococcosis. The review showed that the highest MIC values occurred for voriconazole, especially against C. neoformans. It is concluded that there is a lack of studies with statistical analysis of the data obtained, in order to provide a better dimensioning of the resistance rates of cryptococcosis agents to different antifungal agents, both in geographical and temporal context.

Abstract Background: Cryptoccocal meningitis continues to present high incidence among AIDS patients. The treatment of choice is the synergistic combination of flucytosine (5-FC) with amphotericin B deoxycholate (AmBd) or its lipid formulations. However, 5-FC is unavailable in many countries and AmB demands hospitalization. The combination of AmB with the fungistatic fluconazole (FLC) or the use of high FLC daily doses alone became the choice. Nonetheless, sterilization of cerebrospinal fluid is delayed with FLC monotherapy, mainly with high fungal burden. These findings suggest the search for new antifungal compounds, such as liriodenine. Methods: Liriodenine antifungal activity was evaluated by three procedures: determining the minimum inhibitory concentration (MIC) on 30 strains of the Cryptococcus neoformans (C. neoformans) complex and 30 of the Cryptococcus gattii (C. gattii) complex, using EUCAST methodology and amphotericin B deoxycholate as control; performing the time-kill methodology in two strains of the C. neoformans complex and one of the C. gattii complex; and injury to cryptococcal cells, evaluated by transmission electron microscopy (TEM). Liriodenine absorption and safety at 0.75 and 1.50 mg.kg-1 doses were evaluated in BALB/c mice. Results: Liriodenine MICs ranged from 3.9 to 62.5 μg.mL-1 for both species complexes, with no differences between them. Time-kill methodology confirmed its concentration-dependent fungicidal effect, killing all the strains below the limit of detection (33 CFU.mL-1) at the highest liriodenine concentration (32-fold MIC), with predominant activity during the first 48 hours. Liriodenine induced severe Cryptococcus alterations - cytoplasm with intense rarefaction and/or degradation, injury of organelles, and presence of vacuoles. Liriodenine was better absorbed at lower doses, with no histopathological alterations on the digestive tract. Conclusion: The fungicidal activity confirmed by time-kill methodology, the intense Cryptococcus injury observed by TEM, the absorption after gavage administration, and the safety at the tested doses indicate that the liriodenine molecule is a promising drug lead for development of anticryptococcal agents.

The isolation of Cryptococcosis agents from environmental samples may be difficult due to the presence of groups of fast-growing fungi. We propose a new culture medium based on a modification of Dichloran Rose-Bengal Chloramphenicol Medium (DRBCm) to detect colonies of Cryptococcus neoformans. Our results indicate that DRBCm is superior to the classical Bird Seed Agar in its ability to detect colonies of C. neoformans.

One hundred and forty-one Candida species isolated from clinical specimens of hospitalized patients in Rio de Janeiro, Brazil, during 2002 to 2007, were analized in order to evaluate the distribution and susceptibility of these species to fluconazole. Candida albicans was the most frequent species (45.4%), followed by C. parapsilosis sensu lato (28.4%), C. tropicalis (14.2%), C. guilliermondii (6.4%), C. famata (2.8%), C. glabrata (1.4%), C. krusei (0.7%) and C. lambica (0.7%). The sources of fungal isolates were blood (47.5%), respiratory tract (17.7%), urinary tract (16.3%), skin and mucous membrane (7.1%), catheter (5.6%), feces (2.1%) and mitral valve tissue (0.7%). The susceptibility test was performed using the methodology of disk-diffusion in agar as recommended in the M44-A2 Document of the Clinical and Laboratory Standards Institute (CLSI). The majority of the clinical isolates (97.2%) was susceptible (S) to fluconazole, although three isolates (2.1%) were susceptible-dose dependent (S-DD) and one of them (0.7%) was resistant (R). The S-DD isolates were C. albicans, C. parapsilosis sensu lato and C. tropicalis. One isolate of C. krusei was resistant to fluconazole. This work documents the high susceptibility to fluconazole by Candida species isolated in Rio de Janeiro, Brazil.

Causative agent in majority of VVC is Candida albicans, but infection due to non-C. albicans is common. Use of empiric antifungal therapy in Brazil due to syndromic management of vulvovaginitis could act as risk factor for increase resistance among VVC causative agents. From Mato Grosso patients, 160 with culture-proved among 404 women who had clinical symptoms of VVC, were enrolled in this study. 70 non-pregnant women and 90 pregnant women were included. Candida albicans was the most prevalent, representing 72.9% in the non-pregnant group and 92.3% in the pregnant group. Differences in species distribution were noted between the two groups, being C. parapsilosis the second more prevalent species among non-pregnant women. Susceptibility testing revealed high susceptibility to fluconazole (except for C. krusei), itraconazole, ketoconazole, and amphotericin B regardless the species (C. albicans, C. parapsilosis, C. tropicalis, C. glabrata, C. krusei) analyzed.

FUNDAMENTOS: As micoses superficiais e subcutâneas têm alta prevalência e, muitas vezes, caráter crônico, necessitando tratamentos tópicos e/ou sistêmicos com antifúngicos. As drogas de escolha são azóis e alilaminas (terbinafina). É necessário avaliar a eficácia das drogas para tratamento em humanos e em animais. Estudos para avaliar in vitro a ação dos antimicóticos são raros, especialmente, contra fungos filamentosos. OBJETIVO: Avaliar a eficácia in vitro da terbinafina pelo método de disco-difusão contra fungos filamentosos e leveduras agentes de micoses. MÉTODOS: Avaliou-se a ação da terbinafina (0,125µg-100µg) contra dez espécies fúngicas pelos métodos discodifusão e microdiluição/referência, para determinar a concentração inibitória mínima (MIC). RESULTADOS: Observou-se alta sensibilidade à terbinafina em: T. rubrum, M. gypseum, T. mentagrophytes, T. tonsurans, M. canis, C. carrionii e E. floccosum (halo ≥ 40mm com disco de 0,125µg). S. hyalinum e C. parapsilosis foram considerados sensíveis, mas com halos menores. Fusarium spp. apresentou menor sensibilidade (halo=12mm com disco de 2µg; MIC 8µg/mL). CONCLUSÕES: Os resultados reiteram estudos anteriores quanto à alta eficácia da terbinafina em relação a dermatófitos. A técnica de disco-difusão foi de fácil aplicação e adequada na rotina de laboratórios clínicos.

BACKGROUND: Superficial and subcutaneous mycoses have a high prevalence and, often, chronic evolution. Therefore, they need extensive treatment with topic and/or systemic antifungal agents. Azoles and alilamines (terbinafine) are first-choice drugs to treat human and animal infections. Thus, evaluation of the efficacy of these drugs is important for a successful treatment. However, there are few studies that evaluate the in vitro activity of antifungal agents. OBJECTIVE: To evaluate the in vitro efficacy of terbinafine activity against filamentous fungi and yeasts that cause mycoses. METHOD: The in vitro activity of terbinafine (0.125-100µg) against 10 fungi species was evaluated by the diskdiffusion and microdilution/reference methods to determine the Minimum Inhibitory Concentration (MIC). RESULTS: We found a high susceptibility to terbinafine in: T. rubrum, M. gypseum, T. mentagrophytes, T. tonsurans, M. canis, C. carrionii and E. floccosum (halo ≥ 40mm with 0.125µg disk). S. hyalinum and C. parapsilosis were considered susceptible, but less than the others. Fusarium spp. showed the lowest susceptibility (halo=12mm with 2µg disk; MIC 8µg/mL). CONCLUSIONS: The results of this research confirm previous findings about the efficacy of terbinafine. The drug was shown to be highly effective to treat dermatophyte infections. The disk-diffusion method was easy to use and is a suitable technique for routine use in clinical laboratories.

Natural products have long been providing important drug leads for infectious diseases. Leishmaniasis is a protozoan parasitic disease found mainly in developing countries, and it has toxic therapies with few alternatives. Fungal infections have been the main cause of death in immunocompromised patients and new drugs are urgently needed. In this work, a total of 16 plant species belonging to 11 families, selected on an ethnopharmacological basis, were analyzed in vitro against Leishmania (L.) chagasi, Leishmania (L.) amazonensis, Candida krusei, and C. parapsilosis. Of these plant species, seven showed antifungal activity against C. krusei, five showed antileishmanial activity against L. chagasi and four against L. amazonensis, among them species of genus Plectranthus. Our findings confirm the traditional therapeutic use of these plants in the treatment of infectious and inflammatory disorders and also offer insights into the isolation of active and novel drug prototypes, especially those used against neglected diseases as Leishmaniasis.

This article describes the general objectives of the Regional Laboratory Network for Surveillance of Invasive Fungal Infections and Antifungal Susceptibility in Latin America. Formation of the Network was coordinated by the Essential Medicines, Vaccines, and Health Technologies Unit of the Pan American Health Organization, with the technical and financial support of the National Center for Microbiology of the Carlos III Health Institute (Spain), and the technical support of the Microbiology Department of the Dr. C. Malbrán National Institute on Infectious Diseases (Argentina) and the Microbiology Unit of the Parasitology Service of the Adolfo Lutz Institute (Brazil). The Network's principle objectives are epidemiological surveillance of invasive fungal infections through detection of antifungal resistance and identification of emergent, invasive fungal infections; establishment of norms and common protocols for early diagnosis of mycoses; and strengthening coordination, communications, and transference mechanisms among countries. The Network must be gradually implemented and must include staff training, a systematic process for sharing technology, evaluation of diagnostic techniques, identification of fungal species, and standardized tests for antifungal susceptibility.

Despite highly active anti-retroviral therapy, cryptococcal meningoencephalitis is the second most prevalent neurological disease in Brazilian AIDS patients, being frequently a defining condition with several episodes. As knowledge of Cryptococcus neoformans isolates in the same episode is critical for understanding why some patients develop several episodes, we investigated the genotype characteristics of C. neoformans isolates in two different situations. By pulsed field gel electrophoresis and random amplifield polymorphic DNA analysis, 54 isolates from 12 patients with AIDS and cryptococcosis were analyzed. Group 1 comprised 39 isolates from nine patients with a single episode and hospitalization. Group 2 comprised 15 isolates from three patients with two episodes and hospitalizations. Except for three patients from group 1 probably infected with a single C. neoformans isolate, the other nine patients probably were infected with multiple isolates selected in different collection periods, or the infecting isolate might have underwent mutation to adapt and survive the host immune system and/or the antifungal therapy. However, the three patients from group 2 presented genetic diversity among isolates collected in both hospitalizations, possibly having hosted the initial isolate in both periods. These data, emphasize that Cryptococcus diversity in infection can contribute to strategies of treatment and prevention of cryptococcosis.

The investigation of extracts from six species of marine invertebrates yielded one new and several known natural products. Isoptilocaulin from the sponge Monanchora aff. arbuscula displayed antimicrobial activity at 1.3 mg/mL against an oxacillin-resistant strain of Staphylococcus aureus. Five inactive known dibromotyrosine derivatives, 2 6, were isolated from a new species of marine sponge, Aplysina sp. The sponges Petromica ciocalyptoides and Topsentia ophiraphidites yielded the known halistanol sulfate A (7) as an inhibitor of the antileishmanial target adenosine phosphoribosyl transferase. The ascidian Didemnum ligulum yielded asterubin (10) and the new N,N-dimethyl-O-methylethanolamine (11). The octocoral Carijoa riisei yielded the known 18-acetoxypregna-1,4,20-trien-3-one (12), which displayed cytotoxic activity against the cancer cell lines SF295, MDA-MB435, HCT8 and HL60.

Cryptococcus neoformans, pertencente à classe dos basidiomicetos, é um importante patógeno, principalmente em pacientes imunocomprometidos. Neste estudo, 47 isolados clínicos de C. neoformans de várias regiões do Estado de São Paulo foram avaliados quanto aos sorotipos e ao mating-type por PCR. A diversidade genética foi analisada por PCR-fingerprinting com a seqüência iniciadora específica para regiões microssatélite (GACA)4, RAPD com o iniciador 6 (Amersham Pharmacia Biotech) e por RFLP do gene PLB1 digerido com AvaI. Todos os isolados foram obtidos de pacientes HIV positivos e identificados como sorotipo A e MATalfa. A maioria dos isolados pertencia ao tipo molecular VNI (45/47) e apenas dois foram VNII quando analisados por PCR-fingerprinting e PCR-RFLP. Homogeneidade alta foi obtida com o iniciador (GACA)4, com a maioria dos isolados apresentando correlação alta (> 0.9). Os resultados do RAPD, por sua vez, revelaram maior heterogeneidade com número maior de perfis moleculares. Por PCR-RFLP, nenhum tipo molecular novo foi encontrado, realçando a idéia de que em genes conservados como PLB1, as diferenças podem ser resultantes de divergências evolutivas dentro do complexo C. neoformans, separando os isolados nos oito subtipos moleculares já estabelecidos. Nossos resultados fornecem novas informações sobre a epidemiologia molecular de C. neoformans na região sudeste do Brasil.

The basidiomycetous yeast Cryptococcus neoformans is an important fungal pathogen mainly in immunocompromised patients. In this study, 47 clinical isolates of C. neoformans from regions of São Paulo State were studied serologically by using the Crypto Check Iatron RM 304-K kit, their genetic diversity was estimated by PCR-fingerprinting with a microsatellite-specific sequence (GACA)4, RAPD with primer 6 (Amersham Pharmacia Biotech), PCR-restriction fragment length polymorphism (RFLP) analysis of the phospholipase B gene (PLB1) digested with AvaI and mating type analysis by PCR. All 47 strains isolated from HIV positive patients included in this study were serotype A and MATalpha. The majority of the isolates (45/47) were VNI and only two were VNII by PCR-fingerprinting and PCR-RFLP analysis. High degree of homogeneity was observed when (GACA)4 was used, being highly correlated (> 0.9). In contrast, the RAPD analysis was more heterogeneous with higher number of molecular profiles. By PCR-RFLP, no new molecular type was found, enhancing the suggestion that the differences based on conserved gene as PLB1, can be resultant of ongoing divergent evolution within the C. neoformans complex, into the current eight subtypes. Our results furnish new information on the molecular epidemiology of C. neoformans in the southeast region of Brazil.

Cerebral aspergillosis is a rare cause of brain expansive lesion in AIDS patients. We report the first culture-proven case of brain abscess due to Aspergillus fumigatus in a Brazilian AIDS patient. The patient, a 26 year-old male with human immunodeficiency virus (HIV) infection and history of pulmonary tuberculosis and cerebral toxoplasmosis, had fever, cough, dyspnea, and two episodes of seizures. The brain computerized tomography (CT) showed a bi-parietal and parasagittal hypodense lesion with peripheral enhancement, and significant mass effect. There was started anti-Toxoplasma treatment. Three weeks later, the patient presented mental confusion, and a new brain CT evidenced increase in the lesion. He underwent brain biopsy, draining 10 mL of purulent material. The direct mycological examination revealed septated and hyaline hyphae. There was started amphotericin B deoxycholate. The culture of the material demonstrated presence of the Aspergillus fumigatus. The following two months, the patient was submitted to three surgeries, with insertion of drainage catheter and administration of amphotericin B intralesional. Three months after hospital admission, his neurological condition suffered discrete changes. However, he died due to intrahospital pneumonia. Brain abscess caused by Aspergillus fumigatus must be considered in the differential diagnosis of the brain expansive lesions in AIDS patients in Brazil.

La aspergilosis cerebral es una causa rara de lesión expansiva cerebral en pacientes con SIDA. Presentamos el primer reporte de un absceso cerebral causado por Aspergillus fumigatus en un paciente brasileño con SIDA. El paciente, de 26 años de edad, presentaba antecedentes de infección por el virus de la inmunodeficiencia humana (VIH), tuberculosis pulmonar y toxoplasmosis cerebral. Manifestó fiebre, tos, disnea y dos episódios de convulsiones. La tomografía computadorizada (TC) demostró una lesión hipodensa parasagital y bi-parietal con realce periférico e importante efecto de masa. Se inició tratamiento anti-Toxoplasma. Tres semanas después, el paciente evidenció confusión mental y una nueva TC de cráneo mostró aumento de la lesión. Se realizó biopsia cerebral con drenaje de 10 mL de material purulento. El examen micológico directo reveló hifas hialinas septadas. Se inició anfotericina B deoxicolato. La cultura del material demostró presencia de Aspergillus fumigatus. En los siguientes dos meses el paciente fue sometido a otras tres cirugías, insertándose un catéter de drenaje y administrándose anfotericina B intralesional. Tres meses después de la admisión hospitalaria, la condición neurológica del paciente sufrió discretos cambios. Sin embargo, falleció debido a neumonia intrahospitalaria. Aunque muy raros, los abscesos cerebrales causados por Aspergillus fumigatus deben ser considerados en el diagnóstico diferencial de las lesiones expansivas cerebrales en pacientes con SIDA.

Relata-se o primeiro caso de isolamento de Paracoccidioides brasiliensis (P. brasiliensis) em sangue de paciente HIV positivo, 28 anos, sexo masculino, natural de Nova Londrina que, ao exame físico e ultrassonográfico, apresentava esplenomegalia febril a esclarecer. Para estabelecer o diagnóstico etiológico, hemoculturas em triplicata foram realizadas para pesquisa de bactérias aeróbias, micobactérias e fungos. As hemoculturas para bactérias aeróbias e micobactérias foram negativas e P. brasiliensis foi isolado de duas hemoculturas, na fase leveduriforme em ágar BHI, 20 dias após a semeadura, a partir do meio de Negroni. O paciente classificado, segundo o "Centers for Disease Control (CDC)", no grupo IV devido a uma pneumocistose pulmonar, interrompeu o tratamento por problemas particulares na segunda dose de anfotericina B, sendo tratado alternativamente com 800 mg/dia de cetoconazol. O óbito ocorreu um ano após o isolamento do P.brasiliensis em hemocultura.

A case of an AIDS-patient with positive blood culture for Paracoccidioides brasiliensis is reported. The patient was a 29 years old male born in Nova Londrina (Paraná State, Brazil) who presented splenomegaly and fever of unknown origin. Three blood cultures were performed, each one of them for aerobic bacteria, mycobacteria and fungi. Cultures for aerobic bacteria and mycobacteria were negative. However, the yeast phase P. brasiliensis was isolated from two cultures in BHI agar, 20 days after inoculation in Negroni medium. The patient was classified in group V according to the Centers for Disease Control (CDC) criteria for AIDS, due to a Pneumocystis carinii pneumonia. Treatment was discontinued due to an individual decision of the patient on the second dose of amphotericin B. This antibiotic was replaced by ketoconazole in the daily dose of 800 mg. The patient died one year after the isolation of P. brasiliensis on blood culture.