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Cryptococcal meningitis is a serious infection of the central nervous system that is predominant in developing countries, caused by fungi of the genus Cryptococcus, and which affects immunosuppressed patients, especially those with HIV. Here, we aim to diagnose and characterize the clinical–epidemiological profile of cryptococcosis in patients admitted to two tertiary public hospitals in northeastern Brazil. The study is divided into three moments: (1) the isolation of fungus and diagnosis from biological samples collected between 2017 and 2019, (2) a description of the clinical and epidemiological characteristics of the patients, and (3) the experimental tests related to an in vitro susceptibility antifungal profile. The species were identified by MALDI-TOF/MS. Among the 100 patients evaluated, 24 (24.5%) were diagnosed with cryptococcosis based on positive culture. Clinical–epidemiological analysis showed a slightly higher prevalence in men between 30 and 39 years. When comparing the date of HIV diagnosis and the development of cryptococcosis, it was observed that 50% received the diagnosis of infection by cryptococcosis after or equal to a period of 12 months from being diagnosed with HIV; the other 50% received it within the first 30 days of the HIV diagnosis. Neurocryptococcosis was the most prevalent clinical form, and, at the time of hospital admission, the most common clinical signs were high fever (75%), intense headache (62.50%), and neck stiffness (33.33%). The cerebrospinal fluid showed 100% sensitivity and positivity for direct examination by India ink, and fungal culture. The mortality rate in this study was 46% (11/24), a lower rate than in the other literature. An antifungigram showed that 20 (83.33%) isolates were susceptible to amphotericin B and 15 (62.5%) to fluconazole. Mass spectrometry identified 100% of the isolates as Cryptococcus neoformans. In Brazil, this infection is not mandatory notifiable. Therefore, although there is little information on the subject, it is obsolete and does not express the reality of the facts, mainly in the northeast region, where this information is insufficient. The data obtained in this research contribute to the epidemiological knowledge of this mycosis in Brazil and will serve as a basis for future globally comparative epidemiological studies.

ObjectiveTo describe the clinical–epidemiological features of patients colonized by Candida auris in the largest outbreak in Brazil and to show the biofilm formation capacity of yeast strains.MethodsClinical yeasts suspected of C. auris isolated from urine and surveillance samples were seeded on chromogenic media at 30°C and Sabouraud agar at 42°C. matrix-assisted laser desorption/ionization–time of flight mass spectometry was used for reliable identification. After proteomic confirmation, the genomic approach and culture on Chromagar Candida Plus media were carried out. Biofilm formation was investigated based on metabolic activity, and the clinical–epidemiological profile of patients was described.ResultsA total of 11 C. auris clinical yeasts from nine patients were identified between the end of December 2021 and March 2022. Two clinical yeasts were isolates from urine and nine clinical yeasts were isolates from axillary and inguinal surveillance swabs. No case is related to previous Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. However, all the yeasts showed a high ability of biofilm formation.ConclusionC. auris requires great vigilance as its high capacity to colonize and form biofilms contributes to its dissemination. The rapid and precise identification of this species is essential for the management, control, and prevention of infections.

In the present study we report a case of candidemia in an infant with cystic fibrosis admitted to a Pediatric Intensive Care Unit (PICU) with a fatal course. Six-month-old male patient with a history of productive cough and persistent fever. He was admitted to the PICU of a public hospital in the city of Recife, PE, Brazil, with a diagnosis of dyspnea and cystic fibrosis, with a respiratory infection. Antibiotics were administered, however without success, peripheral blood and transcatheter blood cultures were requested, the isolated agents were Staphylococcus epidermidis and Pseudomonas aeruginosa, respectively. After seven days, the patient presented lesions on the genitalia and persistent fever. Blood and genital lesions were collected, the samples were identified as Candida albicans through the classical taxonomy and the VITEK 120 automated system. The antifungal sensitivity test followed the protocol by the broth microdilution method (CLSI- Clinical and Laboratory Standard Institute, 2008b). The isolates from blood samples were sensitive to amphotericin B with a Minimal Inhibitory Concentration (MIC) of 0.03 µg/mL, 0.12 µg/mL for anidulafungin and 0.25 µg/mL for voriconazole, exhibiting resistance to fluconazole with MIC of 64 µg/mL. C. albicans isolated from the genitalia lesion was sensitive to all the drugs used. The patient was administered nystatin 4 times a day and amphotericin B. The patient improved from the genital lesion and fever. However, after three days, the patient presented cardiac and respiratory deficit that led to cardiac arrest, leading to death. No presente estudo relatamos um caso de candidemia em lactente portador de fibrose cística internado em Unidade de Terapia Intensiva Pediátrica (UTIP) com curso fatal. Paciente com seis meses de idade, do sexo masculino com histórico de tosse produtiva e febre persistente. Foi admitido na UTIP de um hospital público da cidade do Recife, PE, Brasil, com diagnóstico de dispneia e fibrose cística, apresentando um quadro infeccioso respiratório. Foi administrado antibióticos, entretanto sem sucesso, foi solicitada hemocultura de sangue periférico e de transcateter, os agentes isolados foram Staphylococcus epidermidis e Pseudomonas aeruginosa, respectivamente. Após sete dias o paciente apresentou lesão na genitália e febre persistente. Foi realizada coleta de sangue e da lesão da genitália, as amostras foram identificas como Candida albicans através da taxonomia clássica e do sistema automatizado VITEK 120. O teste de sensibilidade a antifúngicos seguiu protocolo pelo método de microdiluição em caldo (CLSI- Clinical and Laboratory Standard Institute, 2008b). Os isolados de amostras de sangue foram sensíveis a anfotericina B com Concentração Inibitória Mínima (CIM) de 0,03 µg/mL, 0,12 µg/mL para anidulafungina e ao 0,25 µg/mL para voriconazol, exibindo resistência ao fluconazol com CIM de 64 µg/mL. C. albicans isolada da lesão da genitália foi sensível a todas as drogas utilizadas. Foi administrado ao paciente nistatina 4 vezes ao dia e anfotericina B. O paciente obteve melhora da lesão da genitália e da febre. Contudo, após três dias o paciente apresentou déficit cardíaco e respiratório que conduziu a parada cardíaca, indo ao óbito. En el presente estudio reportamos un caso de candidemia en un lactante con fibrosis quística ingresado en una Unidad de Cuidados Intensivos Pediátricos (UCIP) con evolución fatal. Paciente masculino de seis meses de edad con antecedentes de tos productiva y fiebre persistente. Ingresó en la UCIP de un hospital público de la ciudad de Recife, PE, Brasil, con diagnóstico de disnea y fibrosis quística, con infección respiratoria. Se administraron antibióticos, sin embargo, sin éxito, se solicitaron hemocultivos de sangre periférica y transcatéter, los agentes aislados fueron Staphylococcus epidermidis y Pseudomonas aeruginosa, respectivamente. Después de siete días, el paciente presentó lesiones en los genitales y fiebre persistente. Se recolectó sangre y lesiones genitales, las muestras se identificaron como Candida albicans mediante la taxonomía clásica y el sistema automatizado VITEK 120. La prueba de sensibilidad antifúngica siguió el protocolo por el método de microdilución en caldo (CLSI- Clinical and Laboratory Standard Institute, 2008b). Los aislados de muestras de sangre fueron sensibles a la anfotericina B con una Concentración Mínima Inhibitoria (MIC) de 0,03 µg/mL, 0,12 µg/mL para anidulafungina y 0,25 µg/mL para voriconazol, presentando resistencia a fluconazol con MIC de 64 µg/mL. C. albicans aislado de la lesión de los genitales fue sensible a todos los fármacos utilizados. Se le administró nistatina 4 veces al día y anfotericina B. La paciente mejoró de la lesión genital y fiebre. Sin embargo, después de tres días, el paciente presentó un déficit cardíaco y respiratorio que lo llevó a un paro cardíaco que lo llevó a la muerte.

Background Opportunistic infections are frequent in people living with the human immunodeficiency virus who either do not have access to antiretroviral therapy (ART) or use it irregularly. Tuberculosis is the most frequent infectious disease in PLHIV and can predispose patients to severe fungal infections with dire consequences. Case presentation We describe the case of a 35-year-old Brazilian man living with human immunodeficiency virus (HIV) for 10 years. He reported no adherence to ART and a history of histoplasmosis with hospitalization for 1 month in a public hospital in Natal, Brazil. The diagnosis was disseminated Mycobacterium tuberculosis infection. He was transferred to the health service in Recife, Brazil, with a worsening condition characterized by daily fevers, dyspnea, pain in the upper and lower limbs, cough, dysphagia, and painful oral lesions suggestive of candidiasis. Lymphocytopenia and high viral loads were found. After screening for infections, the patient was diagnosed with tuberculous pericarditis and esophageal candidiasis caused by Candida tropicalis. The isolated yeasts were identified using the VITEK 2 automated system and matrix-assisted laser desorption/ionization time-of-flight–mass spectrometry. Antifungal microdilution broth tests showed sensitivity to fluconazole, voriconazole, anidulafungin, caspofungin, micafungin, and amphotericin B, with resistance to fluconazole and voriconazole. The patient was treated with COXCIP-4 and amphotericin deoxycholate. At 12 days after admission, the patient developed sepsis of a pulmonary focus with worsening of his respiratory status. Combined therapy with meropenem, vancomycin, and itraconazole was started, with fever recurrence, and he changed to ART and tuberculostatic therapy. The patient remained clinically stable and was discharged with clinical improvement after 30 days of hospitalization. Conclusion Fungal infections should be considered in patients with acquired immunodeficiency syndrome as they contribute to worsening health status. When mycoses are diagnosed early and treated with the appropriate drugs, favorable therapeutic outcomes can be achieved.

Studies on the microbiota of freshwater cetaceans are scarce and may provide important data on animal and environmental health. This study aimed to evaluate the antimicrobial susceptibility of Gram-negative bacteria recovered from two populations of free-ranging Amazon river dolphins (Inia geoffrensis). Twenty-one animals were captured and released, 13 from Negro River and 8 from Tapajos River, Brazil. Swab samples were obtained from the oral cavity, blowhole, genital opening and rectum and were cultured on MacConkey agar. Isolates were biochemically identified, and antimicrobial susceptibility was assessed by disk diffusion method. Overall, 132 isolates were recovered, of which 71 were recovered from animals from Negro River and 61 from Tapajos River. The most commonly recovered bacterial species were Enterobacter cloacae, Morganella morganii, Klebsiella pneumoniae and Pseudomonas aeruginosa. Overall, 51.6% (63/122) of the isolates were not-susceptible (intermediate resistance and resistance), of which 28/122 (22.9%) were resistant to at least one antimicrobial. Cephalothin, cefuroxime and cefepime were the drugs to which more resistant and intermediate results were observed (P

The Brazilian northeastern semi-arid is a region composed of countless microorganisms adapted to extreme conditions, and to ensure their survival, they can produce several biomolecules and develop peculiar mechanisms. The use of microorganisms has been reported as a promising alternative for promoting plant growth through the production of several bioactive molecules, which can replace chemical agents that cause environmental problems. In this study, Bacillus sp. Ar 16 (UFPEDA), isolated from the rhizosphere of Aroeira (Schinus terebinthifolia) from the Caatinga biome, northeastern semi-arid region, was taxonomically characterized through biochemical tests, Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF) and 16S region rRNA such as Bacillus cereus UFPEDA 1060 B (99.86%). In vitro conditions, the B. cereus UFPEDA 1060B strain produced exopolysaccharide (EPS), cellulases, and phosphatases and presented maximum phosphate solubilization of 880 µg.mL -1 . Furthermore, the production of 3.96 to 4.06 µg.mL -1 of indole acetic acid (IAA) in a culture medium supplemented with 5 mM of L-tryptophan was evidenced. IAA and other organic acids were extracted and analyzed by High-Performance Liquid Chromatography (HPLC) at a retention time of 6.2. The studied strain has different mechanisms that can promote plant growth and is also capable of controlling the growth of Fusarium spp. and Colletotrichum spp. The effectiveness of the results presented by this strain demonstrate its biotechnological potential, which can bring benefits to sustainable agriculture with artifices that favor, in addition to plant growth, the cycling of nutrients. La región semiárida nororiental está compuesta por numerosos microorganismos adaptados a condiciones extremas y, para asegurar su supervivencia, pueden producir varias biomoléculas y desarrollar mecanismos peculiares. El uso de microorganismos ha sido reportado como una alternativa promisoria para promover el crecimiento de las plantas através de la producción de varias moléculas bioactivas, que pueden reemplazar a los agentes químicos que causan problemas ambientales. En este estudio, Bacillus sp. Ar 16 (UFPEDA), aislado de la rizósfera de Aroeira (Schinus terebinthifolia) en el bioma Caatinga, región semiárida nororiental, se caracterizó taxonómicamente mediante pruebas bioquímicas, espectrometría de masas de tiempo de vuelo de ionización por desorción láser asistida por matriz (MALDI-TOF) y la región 16S del rRNA como Bacillus cereus UFPEDA 1060 B (99,86%). En condiciones in vitro, la cepa de B. cereus UFPEDA 1060 B produjo exopolisacáridos (EPS), celulasas, fosfatasas y presentó una solubilización máxima de fosfato de 880 µg.mL-1. Además, se evidenció la producción de 3.96 a 4.06 µg.mL-1 de ácido indol acético (IAA) en medio de cultivo suplementado con 5 mM de L-triptófano. El IAA y otros ácidos orgánicos se extrajeron y analizaron mediante cromatografía líquida de alta resolución (HPLC) con un tiempo de retención de 6,2. La cepa estudiada tiene diferentes mecanismos que pueden promover el crecimiento de las plantas y también es capaz de controlar el crecimiento de Fusarium spp. y Colletotrichum spp. La efectividad de los resultados presentados por esta cepa demuestra su potencial biotecnológico, que puede traer beneficios a la agricultura sustentable con artificios que favorecen, además del crecimiento de las plantas, el ciclo de nutrientes. O semiárido nordestino é uma região composta por inúmeros microrganismos adaptados a condições extremas e, para garantir sua sobrevivência, podem produzir diversas biomoléculas e desenvolver mecanismos peculiares. A utilização de microrganismos tem sido relatada como uma alternativa promissora para promover o crescimento de plantas através da produção de diversas moléculas bioativas, que podem substituir agentes químicos que causam problemas ambientais. Neste estudo, Bacillus sp. Ar 16 (UFPEDA), isolado da rizosfera da Aroeira (Schinus terebinthifolia) do bioma Caatinga, semiárido nordestino, foi caracterizado taxonomicamente através de testes bioquímicos, Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF) e região 16S do rRNA como Bacillus cereus UFPEDA 1060 B (99,86%). Em condições in vitro, a cepa de B. cereus UFPEDA 1060B produziu exopolissacarídeos (EPS), celulases, fosfatases e apresentou solubilização máxima de fosfato de 880 µg.mL-1. Além disso, foi evidenciada a produção de 3,96 a 4,06 µg.mL-1 de ácido indol acético (AIA) em meio de cultura suplementado com 5 mM de L-triptofano. O AIA e outros ácidos orgânicos foram extraídos e analisados ​​por Cromatografia Líquida de Alta Eficiência (CLAE) em um tempo de retenção de 6,2. A linhagem estudada possui diferentes mecanismos que podem promover o crescimento das plantas e também é capaz de controlar o crescimento de Fusarium spp. e Colletotrichum spp. A eficácia dos resultados apresentados por esta linhagem demonstra seu potencial biotecnológico, que pode trazer benefícios à agricultura sustentável com artifícios que favorecem, além do crescimento das plantas, a ciclagem de nutrientes.

Background: Sporotrichosis occurs through contact with contaminated soil and plant. However, the incidence of sporotrichosis as a zoonotic epidemic has increased, particularly in Rio de Janeiro. Aim: In this work, we decided to evaluate some T-cell phenotypes involved in the immune response. Materials & methods: We used flow cytometry to quantify TCD4+ and TCD8+ and Treg from immunocompetent and immunosuppressed mice infected with Sporothrix species with different levels of virulence and pathogenicity. Results: It was demonstrated the predominance of TCD4+ over the TCD8+ cells in both groups, inoculated with all the species, and percentages of Treg observed in infected immunocompetent mice. Conclusion: This regulatory phenotype can be associated with a protective immunity in the initial periods of infection.

Sporotrichosis is a subcutaneous mycosis caused by thermodimorphic fungi of Sporothrix schenckii complex and is usually transmitted as a result of traumatic inoculation of these microorganisms; infection can produce a number of manifestations. 1,2 In Brazil, sporotrichosis has been a major public health issue, with multiple outbreaks occurring because of zoonotic transmission, including contact with cats, and S brasiliensis is the most prevalent agent in Rio de Janeiro State.3,4 Immunocompromised individuals are thought to be at increased risk of developing a more severe clinical presentation.5 Disseminated sporotrichosis in immunocompetent individuals as result of contact with cats has becoming increasingly reported. 6,7 Historically, cases of sporotrichosis in the Brazilian northeast have been infrequently reported, and this was not thought to be an endemic area for this mycosis. Herein, we report 4 cases of disseminated sporotrichosis in patients in a public hospital in Pernambuco. Publication of this case series report was granted institutional review board approval by the Ethics Committee in Research at the Federal University of Pernambuco, Brazil.

The objective of this study was to evaluate the effects of nanoparticles (nanospheres and nanocapsules) of the promising antifungal 2-amino-thiophene (6CN10) and 6CN10 complexed with 2-hydroxypropyl-β-cyclodextrin (6CN10:HP-β-CD) in vitro and compared with free drug against Candida and Cryptococcus, using a microdilution method to measure susceptibility. The Candida and Cryptococcus clinical strains were identified using phenotypic methods and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). To measure in vitro antifungal susceptibility, we used microdilution trials. Serial drug or nanoparticle dilutions were prepared according to the CLSI M27-A3 guidelines. Anti-biofilm activity was verified for Cryptococcus neoformans. All Candida isolates were sensitive to the free drug (MIC = 41.66-333.33 μg/mL) and were able to grow even at the higher concentration tested for all 6CN10 nanoparticles. However, the Cryptococcus neoformans strains presented MIC values of 0.32-83.33 μg/mL for 6CN10 nanoparticles, and MIC values of 0.1-0.2 μg/mL for 6CN10:HP-β-CD nanoparticles, i.e., 3333 times more active than the free drug (MIC values 166.66-333.33 μg/mL), and presenting activity greater than that of the reference drug amphotericin B (MIC = 0.5-0.125 μg/mL). 6CN10:HP-β-CD nanosphere also showed high anti-biofilm potential. The in vitro study showed that the nanoparticles allowed better drug efficiency against Cryptococcus than did the free drug. These results suggest that 6CN10-loaded nanoparticles may become a future alternative for cryptococcosis and candidiasis therapy. In vivo experiments are essential prior to clinical use.

The leaves of P. edulis were subjected to physicochemical analysis, such as ion content, extractives, and structural molecules. The hexanic, ethanolic and ethyl acetate extracts were submitted to phytochemical analyzes by GC-MS, HPLC-MS, and spectrophotometry. In addition, antioxidant (DPPH, ABTS and TAA methods) potential, antimicrobial (MIC method) action, cytotoxicity and immunostimulant activity (flow cytometry analysis) were performed. The extracts showed a moderate antioxidant capacity and revealed the presence of several metabolites, mainly phenols, such as caffeic acid, p-coumaric acid and luteolin. The ethyl acetate and ethanolic extracts showed antifungal activity. In addition, the extracts did not affect splenocytes viability at 12.5 μg/mL and promoted the production of IL-6, IL-10, IL-17 and TNF-α cytokines. P. edulis extracts showed antifungal and antioxidant activity and were able to induce immunostimulatory action in splenocyte cultures in vitro.

Aim: Cases of sporotrichosis are emerging in several states of Brazil, especially in the southeast. Recently, sporotrichosis has been reported in the state of Pernambuco in the northeastern region. The goal of this study was to shed new light on sporotrichosis in terms of the geographic distribution of human cases and provide an overview of sporotrichosis associated with zoonotic transmission. Patients & methods: From March 2017 to November 2019, 179 patients were diagnosed with sporotrichosis. Georeferencing analysis, spatial distribution and epidemiological features of all cases are described. Results: The data show the dynamics of accelerated transmission of sporotrichosis across urban and coastal areas of the state of Pernambuco. Conclusion: There is a need to decentralize health services and implement a One Health approach to this emerging disease.

Psoriasis is a chronic inflammatory disease that affects over 125 million people worldwide. Many studies have shown the importance of the microbiome for psoriasis exacerbation.Explore the fungal load and species composition of cultivable yeasts on the skin of psoriatic patients (PP) and healthy volunteers living in a tropical area and evaluate the susceptibility to antifungals.A cross-sectional study with 61 participants (35 patients and 26 healthy controls) was performed during August 2018 and May 2019. Clinical data were collected from patient interviewing and/or medical records review. Samples were collected by swabbing in up to five anatomic sites. Suggestive yeast colonies were counted and further identified by phenotypical tests, PCR-REA, and/or MALDI-TOF. Susceptibility of Malassezia spp. and Candida spp. to azoles, terbinafine, and amphotericin B was evaluated by broth microdilution.Nearly 50% of the patients had moderate to severe psoriasis, and plaque-type psoriasis was the most common clinical form. Yeast colonies count was significantly more abundant among PP than healthy controls. Malassezia and Candida were the most abundant genus detected in all participants. Higher MIC values for ketoconazole and terbinafine were observed in Malassezia strains obtained from PP. Approximately 42% of Candida isolates from PP showed resistance to itraconazole in contrast to 12.5% of isolates from healthy controls. MIC values for fluconazole and amphotericin B were significantly different among Candida isolates from PP and healthy individuals.This study showed that Malassezia and Candida strains from PP presented higher MIC values to widespread antifungal drugs than healthy individuals.

Cryptococcosis is an infectious disease of worldwide distribution, caused by encapsulated yeasts belonging to the phylum Basidiomycota. The genus Cryptococcus includes several species distributed around the world. The C. gattii/neoformans species complex is largely responsible for most cases of cryptococcosis. However, clinical series have been published of infections caused by Papiliotrema (Cryptococcus) laurentii and Naganishia albida (Cryptococcus albidus), among other related genera. Here, we examined the pathogenic potential and antifungal susceptibility of C. gattii/neoformans species complex (clades I and II) and related genera (Papiliotrema and Naganishia) isolated from environmental and clinical samples. P. laurentii (clade III), N. liquefasciens/N. albidosimilis (clade IV); and N. adeliensis/N. albida (clade V) strains produced higher levels of phospholipase and hemolysins, whereas the C. gattii/neoformans species complex strains (clades I and II) had markedly thicker capsules, produced more biofilm biomass and melanin, which are known virulence attributes. Interestingly, 40% of C. neoformans strains (clade II) had MICs above the ECV established for this species to amphotericin B. Several non-C. gattii/neoformans species complex (clades III to V) had MICs equal to or above the ECVs established for C. deuterogattii and C. neoformans for all the three antifungal drugs tested. Finally, all the non-C. gattii/neoformans clinical isolates (clades III to V) produced more melanin than the environmental isolates might reflect their particularly enhanced need for melanin during in vivo protection. It is very clear that C. gattii/neoformans species complex (clades I and II) strains, in general, show more similar virulence phenotypes between each other when compared to non-C. gattii/neoformans species complex (clades III to V) isolates. These observations together with the fact that P. laurentii and Naganishia spp. (clades III to V) strains were collected from the outside of a University Hospital, identify features of these yeasts important for environmental and patient colonization and furthermore, define mechanisms for infections with these uncommon pathogens.

Persister cells are metabolically inactive dormant cells that lie within microbial biofilms. They are phenotypic variants highly tolerant to antimicrobials and, therefore, associated with recalcitrant infections. In the present study, we investigated if Trichosporon asahii and T. inkin are able to produce persister cells. Trichosporon spp. are ubiquitous fungi, commonly found as commensals of the human skin and gut microbiota, and have been increasingly reported as agents of fungemia in immunocompromised patients. Biofilms derived from clinical strains of T asahii (n=5) and T. inkin (n=7) were formed in flat-bottomed microtiter plates and incubated at 35°C for 48 h, treated with 100 μg/ml amphotericin B (AMB) and incubated at 35°C for additional 24 h. Biofilms were scraped from the wells and persister cells were assayed for susceptibility to AMB. Additionally, we investigated if these persister cells were able to generate new biofilms and studied their ultrastructure and AMB susceptibility. Persister cells were detected in both T asahii and T. inkin biofilms and showed tolerance to high doses of AMB (up to 256 times higher than the minimum inhibitory concentration). Persister cells were able to generate biofilms, however they presented reduced biomass and metabolic activity, and reduced tolerance to AMB, in comparison to biofilm growth control. The present study describes the occurrence of persister cells in Trichosporon spp. and suggests their role in the reduced AMB susceptibility of T. asahii and T. inkin biofilms.

Chlamydoconidium-producing Trichophyton tonsurans strains isolated in Northeastern Brazil have morphological features different from the classic description of this dermatophyte species. This study investigated the phylogenetic relationship of chlamydoconidium-producing T. tonsurans strains isolated in Northeastern Brazil. Also, the effect of terbinafine and farnesol on mature biofilms of T. tonsurans strains was evaluated. The mass spectra of T. tonsurans strains were investigated by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The ITS and LSU loci regions of rDNA and the partial β-tubulin gene were sequenced and the phylogenetic tree was analysed. The effects of terbinafine and farnesol on mature T. tonsurans biofilms were evaluated through the analysis of metabolic activity, quantification of biomass and observation by scanning electron microscopy. MALDI-TOF MS spectra of the chlamydoconidium-producing T. tonsurans strains differed from the spectrum of the control strain (ATCC 28942), presenting an intense ion peak at m/z 4155 Da. Phylogenetic tree analysis showed that the chlamydoconidium-producing strains isolated in Northeastern Brazil are allocated to a single cluster, differing from strains isolated from other countries. As for mature T. tonsurans biofilms, farnesol reduced biomass and metabolic activity by 64.4 and 65.9 %, respectively, while terbinafine reduced the biomass by 66.5 % and the metabolic activity by 69 %. Atypical morphological characteristics presented by chlamydoconidium-producing T. tonsurans strains result from phenotypic plasticity, possibly for adaptation to environmental stressors. Also, farnesol had inhibitory activity against T. tonsurans biofilms, demonstrating this substance can be explored for development of promising anti-biofilm drugs against dermatophytes.

Cryptococcosis is an infectious disease of worldwide distribution, caused by encapsulated yeasts belonging to the phylum Basidiomycota. The genus Cryptococcus includes several species distributed around the world. The C. gattii/neoformans species complex is largely responsible for most cases of cryptococcosis. However, clinical series have been published of infections caused by Papiliotrema (Cryptococcus) laurentii and Naganishia albida (Cryptococcus albidus), among other related genera. Here, we examined the pathogenic potential and antifungal susceptibility of C. gattii/neoformans species complex (clades I and II) and related genera (Papiliotrema and Naganishia) isolated from environmental and clinical samples. P. laurentii (clade III), N. liquefasciens/N. albidosimilis (clade IV); and N. adeliensis/N. albida (clade V) strains produced higher levels of phospholipase and hemolysins, whereas the C. gattii/neoformans species complex strains (clades I and II) had markedly thicker capsules, produced more biofilm biomass and melanin, which are known virulence attributes. Interestingly, 40% of C. neoformans strains (clade II) had MICs above the ECV established for this species to amphotericin B. Several non-C. gattii/neoformans species complex (clades III to V) had MICs equal to or above the ECVs established for C. deuterogattii and C. neoformans for all the three antifungal drugs tested. Finally, all the non-C. gattii/neoformans clinical isolates (clades III to V) produced more melanin than the environmental isolates might reflect their particularly enhanced need for melanin during in vivo protection. It is very clear that C. gattii/neoformans species complex (clades I and II) strains, in general, show more similar virulence phenotypes between each other when compared to non-C. gattii/neoformans species complex (clades III to V) isolates. These observations together with the fact that P. laurentii and Naganishia spp. (clades III to V) strains were collected from the outside of a University Hospital, identify features of these yeasts important for environmental and patient colonization and furthermore, define mechanisms for infections with these uncommon pathogens.

Biosensors are pre-prepared diagnostic devices composed of at least one biological probe. These devices are envisaged for the practical identification of specific targets of microbiological interest. In recent years, the use of narrow-specific probes such as lectins has been proven to distinguish bacteria and glycoproteins based on their superficial glycomic pattern. For instance, Concanavalin A is a carbohydrate-binding lectin indicated as a narrow-specific biological probe for Gram-negative bacteria. As a drawback, Gram-positive bacteria are frequently overlooked from lectin-based biosensing studies because their identification results in low resolution and overlapped signals. In this work, the authors explore the effect that platform nanostructuration has over the electrochemical response of ConA-based platforms constructed for bacterial detection; one is formed of chitosan-capped magnetic nanoparticles, and another is composed of gold nanoparticle-decorated magnetic nanoparticles. The biosensing platforms were characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) as a function of bacterial concentration. Our results show that probe-target interaction causes variations in the electrical responses of nanostructured transducers. Moreover, the association of gold nanoparticles to magnetic nanoparticles resulted in an electrical enhancement capable of overcoming low resolution and overlapping Gram-positive identification. Both platforms attained a limit of detection of 10 ° CFU mL−1, which is useful for water analyses and sanitation concerns, where low CFU mL−1 are always expected. Although both platforms were able to detect Gram-negative bacteria, Gram-positives were only correctly differentiated by the gold nanoparticle-decorated magnetic nanoparticles, thus demonstrating the positive influence of hierarchically nanostructured platforms.

Chlamydoconidium-producing

Coccidioidomycosis is a deep-seated fungal infection that occurs exclusively in semiarid areas in the Americas. In Brazil, coccidioidomycosis occurs exclusively in rural areas in the northeast region and affects counties that are hit by recurrent droughts, poverty and economic stagnation. Since 1978, approximately 136 cases of the disease have been reported in Brazil, according to scientific publications. However, a lack of governmental epidemiological data as well as a similarity to tuberculosis have led scientists and experts to assume that a greater number of cases occur in the country, which are not diagnosed and/or reported. In this review, general characteristics of coccidioidomycosis are presented, followed by a description of the main clinical and epidemiological data of cases in Brazil. The purpose of this article is to discuss the inclusion of coccidioidomycosis in the list of neglected tropical diseases. We believe that the adoption of coccidioidomycosis as a neglected tropical disease will enable the creation of an effective epidemiological surveillance system and the development of feasible public health solutions for its control in vulnerable populations.

Seventeen Schiff bases bearing 2-aminothiophene derivatives were designed and synthesized using molecular simplification. The resulting compounds (4a-4q) were evaluated for their in vitro antifungal activity against dermatophytes. Prediction of their druglikeness and pharmacokinetic properties, establishment of their structure-activity relationships (SAR), and cytotoxic evaluation of the most active compounds were performed. Using an eco-friendly procedure, microwave assisted synthesis resulted in compounds in good yields (35-85%). Compounds 4a-4q presented good druglikeness and pharmacokinetic profiles and no cytotoxicity for any cell line tested up to 100 μM. The compounds presenting the best antifungal profiles were 4e, 4f, 4g, 4k, 4l, 4m, 4o and 4p with more than one minimum inhibitory concentration (MIC) value occurring between 16‑64 μg mL-1, thus, in some cases better than the reference drug (fluconazole). SAR testing indicated that the presence of halogens and nitro substituents increases antifungal activity. Taken together, the results demonstrate that 2-aminothiophene derivatives are promising lead compounds for the development of antifungal drugs.

Giardia lamblia e um dos principais protozoarios que causa diarreia em criancas de paises em desenvolvimento, assim esse trabalho teve como objetivo avaliar a prevalencia de infeccao por Giardia sp nas criancas de uma creche publica do Recife, correlacionando os aspectos antropometricos e socioeconomicos, tais como: tabagismo na familia, aleitamento materno, renda e escolaridade materna. A frequencia foi determinada atraves de amostras fecais processadas de acordo com os metodos de Hoffman e Faust. Criancas com infeccao por outros parasitos foram eliminadas do estudo. A prevalencia foi correlacionada com aspectos antropometricos e sociais. A analise desses dados mostrou haver associacao estatisticamente significante da infeccao por Giardia sp com os baixos indices de IMC (p=0,0293) e com o baixo grau de aleitamento materno (p=0,0447), alem de mostrar associacao parcialmente significante com a baixa renda familiar (p=0,09). Os resultados sugerem haver associacao entre a infeccao e desnutricao e aleitamento materno inadequado (por um periodo menor que 6 meses). A renda familiar sugere uma forte tendencia a infeccao nas classes sociais mais baixas.

Human sporotrichosis is an emerging disease caused by fungi of the genus Sporothrix, distributed worldwide, but mostly in tropical and subtropical regions. This disease is caused by traumatic inoculation of contaminated material (either animal or vegetal in origin) into the skin. Sporotrichosis cases caused by zoonotic transmission through felines have significantly increased over the last 20 years in Brazil. There is a spectrum of clinical outcomes, from classical lymphocutaneous and fixed forms to disseminated manifestations and extracutaneous lesions; however, hypersensitivity reactions related to sporotrichosis, including Sweet syndrome (acute febrile neutrophilic dermatoses), are uncommon. In Brazil, Sporothrix brasiliensis is repeatedly associated with feline infection and has consistently shown higher virulence, tendency to escalate to outbreaks or epidemics, and development of atypical forms. Therefore, the objective of the present study was to report the cases of 10 patients with sporotrichosis infected by S. brasiliensis species who developed Sweet syndrome to alert this association, especially in endemic areas.

This study proposes a microcosm biofilm (MiB) model for the study of vulvovaginal candidiasis (VVC). Different conditions that mimic the vaginal environment were tested for MiB formation. The best growth conditions were obtained with samples incubated in vaginal fluid simulator medium pH 4.5 at 35 °C under a microaerophilic atmosphere. MiBs were evaluated for growth kinetics, fluconazole susceptibility and morphology. Samples containing high numbers of bacteria were analyzed for metagenomics. At 48 h, MiBs presented a higher cell density (CFU ml

The aim of this preliminary study was to identify microorganisms with antimicrobial resistance profile and biofilm producers in oropharynx of Rupornis magnirostris and Caracara plancus. Six R. magnirostris and six C. plancus maintained in Triage Center for Wild Animals (CETAS) facilities were studied. Coagulase-positive staphylococci (CoPS), enterobacteria, and yeasts were identified by the biochemical analysis or MALDI-TOF mass spectrometry. The resistance profile of the microorganisms was analyzed according to CLSI. The biofilm production was evaluated by Congo red and violet crystal staining methods. Among the 12 birds, 10 presented strains of CoPS and/or enterobacteria with resistance profile, such as methicillin-resistant CoPS (MR-CoPS), vancomycin-resistant CoPS (VR-CoPS), extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL), and Klebsiella pneumoniae carbapenemase- (KPC-) producing bacteria. Regards the fungal analysis, Candida spp., Cryptococcus spp., Rhodotorula mucilaginosa, R. glutinis, and Trichosporon coremiiforme were identified. All the Trichosporon coremiiforme strains were resistant to amphotericin B, as well as all the Rhodotorula mucilaginosa exhibited resistance to fluconazole. Related to the biofilm production, among the 8 CoPS, 27 enterobacteria, and 10 yeasts isolates, 3, 16, and 7 strains were biofilm producers, respectively. Thus, the presence of these microorganisms in birds of prey is worrisome, highlighting its possible influence in the spread of infections in urban centers.

Caesalpinia pulcherrima is a shrub with worldwide distribution used as an ornamental plant. In this study, we extracted a lignin from the C. pulcherrima leaves and investigated its biological functions. The lignin was characterized by FT-IR, UV–Vis, GPC, TGA and nuclear magnetic resonance (1H and 13C). The antioxidant activity was evaluated using phosphomolybdenum complexation methods (TAA), sequestration of DPPH and ABTS radicals, reducing power, formation of nitrite radical and iron chelating activity (Fe2 +). Antifungal activity was made using Candida spp., Aspergillus spp. and Cryptococcus neoformans strains. Cytotoxicity, oxidative stress, and cytokine production were performed using mouse splenocytes. The lignin showed maximal UV–Vis at ~280 nm, 22.27 L/g·cm of absorptivity and, 2,503 kDa of molecular weight. Phenolic compounds (41.33 ± 0.65 mg GAE/g) and indications of a guaiacyl-syringyl-hydroxyphenyl (GSH)-type composition were found. Antioxidant activities of lignin to TAA (40±1.2%) and to DPPH (16.9±0.2%) was high and showed antifungal potential, especially against Candida spp. (IC50 = 31.3 μg/mL) and C. neoformans (15.6 μg/mL). In mouse splenocytes, the lignin was not cytotoxic and stimulated the cell proliferation and cytokine release. These results indicate that C. pulcherrima lignin has the potential to be used as antifungal and immunostimulant compound.

Fungi stand out as primary pathogens present in healthcare-acquired infections, presenting an increased number of cases even using appropriate antifungal therapy. Candida spp. is a predominant microorganism among several fungal pathogens present in the healthcare setting. Candidemia and candidiasis are fungal infections responsible for high morbidity and mortality among ill patients in hospitals. It is noticeable that prolonged hospital stays lead to a higher economic impact and increased risk for developing secondary fungal or even bacterial infections. New fast and sensitive approaches for the detection of Candida species is highly required. Electrochemical biosensors are an excellent alternative to conventional techniques by combining fast analyte detection, low cost, and the possibility of miniaturization. Lectins are carbohydrate-binding proteins with the capability to reach out to the microorganism cell wall. In this work, we proposed the development of an impedimetric biosensor for Candida spp. based on Concanavalin A (ConA) and wheat germ agglutinin (WGA) as recognition agents of the yeast cells. Atomic force microscopy images indicate changes in the biosensor surface after assembly of the molecules and exposure to fungal samples. Electrochemical impedance spectroscopy results revealed a proportional increase of charge transfer resistance (RCT) as fungal CFU increased, where four Candida species were evaluated (Candida krusei, Candida tropicalis, Candida parapsilosis and Candida albicans). The biosensor is useful to differentiate Candida spp. with a detection limit between 102 to 106 CFU mL−1. The obtained biosensor appears as an innovative candidate for the detection and differentiation of pathogenic Candida spp.

Sporothrix chilensis is a mild-pathogenical specie of Sporothrix pallida complex, until now, known as restrict to Chile. Herein, we describe the first clinical isolates identified as S. chilensis in Brazil, preserved in the URM Culture Collection, by polyphasic taxonomy, and their respective antifungal profile of this emergent fungus., Author summary Fungi of the Sporothrix schenckii Complex are the causative agents of the sporotrichosis, subcutaneous mycosis that presents a massive global distribution. In this work, we perform a taxonomic review, by a poliphasic approach, of Sporothrix sp. isolates stored in the University of Recife Culture Collection (Micoteca URM), previously identified as Sporothrix schenckii by classic methods of taxonomic identification. In addition, we perform the method of broth microdilution, aiming to acquire the minimal inhibitory concentration of the standard antifungals (itraconazole and amphotericin B) against these isolates. By sequencing of the partial β-tub-encoding gene, we confirm the occurrence of S. schenckii, and identified four isolates as the S. pallida Complex component, S. chilensis. All the isolates were susceptible to the antifungals, except one, that shows resistance to amphotericin B. Our study demonstrated the first occurrence of S. chilensis in Brazil. Also, we provide the first data concerned the resistance profile of this specie for the choice antifungals in the sporotrichosis treatment, what will have important implications with this regard in the future.

Mass spectrometry by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was used to identify and differentiate the pattern of susceptibility of clinical isolates of Candida parapsilosis complex. 17 C. parapsilosis sensu stricto, 2 C. orthopsilosis, and 1 C. metapsilosis strains were obtained from blood cultures, and three different inocula (103, 105, and 107 CFU/mL) were evaluated against three echinocandins at concentrations ranging from 0.03 to 16 &micro, g/mL after incubation of 1 h, 2 h, and 3 h. Drug-free control was used. The spectra obtained at these concentrations were applied to generate composite correlation index (CCI) matrices for each yeast individually. After cross correlations and autocorrelations of each spectra with null (zero) and maximal (16) concentrations, the CCI was used as separation parameter among spectra. Incubation time and inoculum were critical factors to reach higher precision and reliability of this trial. With an incubation time of 3 h and inoculum of 107 CFU/mL, it was possible to determine the breakpoint of the clinical yeasts by MALDI-TOF that presented high agreement with the clinical laboratory standard institute (CLSI) reference method. Herein, we show that mass spectrometry using the MALDI-TOF technique is powerful when it exploits antifungal susceptibility testing assays.

Mycobacterium tuberculosis, which is responsible for tuberculosis (TB) and Cryptococcus sp. responsible for cryptococcosis, are pathogenic microorganisms that especially affect patients infected with the human immunodeficiency virus (HIV). Both diseases present similar classic symptoms, which makes diagnosis and treatment consequently difficult. To our knowledge, a few reported cases of M. tuberculosis and Cryptococcus sp. co-infection in non-HIV patients exist. This study reports a TB and neurocryptococcosis (NC) comorbidity case in a patient who had no clinical or serological evidence of HIV-compromised immunity. A 49-year-old male patient, a farmer with a low education level, previously diagnosed with TB and was undergoing treatment for a month when he presented progressive headaches, fever, drowsiness and photosensitivity, a stiff neck and a positive Lasegue test. During hospitalization, the patient was also diagnosed with NC through cerebrospinal fluid (CSF) analysis, which revealed the presence of capsulated yeasts by contrast with india ink. Following the yeast isolation, proteomic and molecular analyzes were performed. The patient received antifungal therapy in parallel with TB treatment, which caused complications and had to be modified twice. However, after three months of hospitalization the patient was discharged. Tuberculosis and cryptococcosis co-infection is a clinical and laboratory challenge, often leading to a delay in diagnosis. In this paper we emphasize the need to understand these infectious comorbidities in non-HIV patients from South America, since the few cases reported in the literature are from studies conducted in the United States and China.

A Fusariose Cutânea é uma micose oportunista emergente causada por fungos do gênero Fusarium spp. É transmitida, em regra, pela inalação de aerossóis de conídios fusariais, podendo ainda ser adquirida por implantação traumática em pele. Acomete, principalmente, pacientes com neoplasias hematológicas e receptores de transplante de medula óssea, em quem causa doença disseminada grave. Em imunocompetentes, é responsável por quadros localizados, especialmente cutâneos. O objetivo deste trabalho é descrever um relato de caso de Fusariose Cutânea localizada em indivíduo imunocompetente, adquirida através de implantação traumática. O caso foi observado em um paciente do sexo masculino, 49 anos, agricultor em plantação de banana, sem comorbidades prévias, que procurou o ambulatório de Dermatologia do Hospital Universitário Oswaldo Cruz, Recife/Pernambuco - Brasil, devido ao surgimento de lesões ulceradas de bordas eritematosas infiltradas com centro necrótico em região de lábio inferior, região supramentoniana e terço distal de sobrancelha esquerda com início em novembro/2018. Foram realizadas biópsias das lesões, cujo histopatológico evidenciou dermatite granulomatosa tuberculoide com pequenos focos abscedados. Como o paciente era proveniente de area endêmica para Leishmaniose Cutânea, foi optado por tratamento da patologia com Antimoniato de Meglumina intravenosa por 50 dias, iniciado em junho/2019. O paciente retornou em dezembro, com piora nas lesões supramentoniana e em sobrancelha, além de surgimento de hepatoesplenomegalia. Devido à severidade do caso, foi optado por internamento hospitalar com realização de novas biópsias e início de Anfotericina B Desoxicolato. Realizada testagem para HIV com resultado negativo, Histopatológico da biópsia da lesão de mento evidenciou filamentos miceliais septados e hialinos por microscopia e fungo filamentoso foi isolado em Sabouraud agar após 5 dias de cultivo. Realizado MALDI-TOF/MS na amostra com identificação de Fusarium solani sensível a anfotericina B e itraconazol e com sensibilidade dose dependente a cetoconazol e voriconazol. Voriconazol 200 mg duas vezes ao dia foi iniciado com melhora importante nas lesões de pele. Paciente recebeu alta com regressão quase total das lesões e assintomático. O esquema foi trocado para Itraconazol por maior disponibilidade da medicação, completando 3 semanas de tratamento com triazólicos. Paciente retornou à consulta ambulatorial em janeiro/2020 com regressão completa das lesões.