Impact on the clinical evolution of patients with COVID-19 from an Intensive Care Unit with isolation of Candida spp. in respiratory samples / Impacto na evolução clínica de pacientes com COVID-19 de uma Unidade de Terapia Intensiva com isolamento de Candida spp. em amostras respiratórias / Impacto en la evolución clínica de pacientes con COVID-19 de una Unidad de Cuidados Intensivos con aislamiento de Candida spp. en muestras respiratorias

Background and Objectives: several patients with COVID-19 require hospital admission due to severe respiratory complications and undergo intensive care with mechanical ventilation (MV) support. Associated with this situation, there is an increase in fungal co-infections, which has a negative impact on the outcome of COVID-19. In this regard, this study intended to compare Candida spp. incidence in the respiratory tract of patients admitted in the COVID and General Intensive Care Units (ICU) at a teaching hospital in 2021. Methods: the results of protected tracheal aspirate samples from 556 patients admitted to the COVID ICU and 260 to General ICU as well as the respective records. Results: of the patients analyzed, 38 revealed a positive sample for Candida in the COVID ICU and 10 in the General ICU, with an incidence of 68.3/1000 and 38.5/1000, respectively. Males were predominant in both wards. The most affected age group was the population over 60 years old, and the average hospital admission for the COVID ICU was 22.1 years, and for the General ICU, 24.2. Conclusion: Candida albicans was the most frequently isolated species, and the mortality rate in patients positive for Candida was higher in patients with COVID-19 compared to patients in the General ICU, suggesting that patients infected with SARS-CoV-2, admitted to the ICU under MV, are more predisposed to colonization by Candida spp., which can have a fatal outcome in these patients.(AU)

Justificativa e objetivos: muitos pacientes com COVID-19 necessitam de hospitalização devido às complicações respiratórias graves, e são submetidos a cuidados intensivos com suporte de ventilação mecânica (VM). Associado a esse quadro, verifica-se o aumento de coinfecções fúngicas, que tem impacto negativo no desfecho da COVID-19. Nesse sentido, este estudo pretendeu comparar a incidência de Candida spp. no trato respiratório de pacientes internados nas Unidades de Terapia Intensiva (UTI) COVID e Geral em um hospital escola em 2021. Métodos: foram avaliados os resultados de amostras de aspirado traqueal protegido provenientes de 556 pacientes internados na UTI COVID e 260 na UTI Geral, bem como os respectivos prontuários. Resultados: dos pacientes analisados, 38 revelaram amostra positiva para Candida na UTI COVID e 10 na UTI Geral, com incidência de 68,3/1000 e 38,5/1000, respectivamente. O sexo masculino foi predominante em ambas as alas. A faixa etária mais acometida foi a população acima de 60 anos, e a média de internação para a UTI COVID foi de 22,1 anos, e para a UTI Geral, 24,2. Conclusão: Candida albicans foi a espécie isolada com maior frequência, e a taxa de mortalidade em pacientes com positivos para Candida foi maior em pacientes com COVID-19 em relação aos pacientes da UTI Geral, sugerindo que pacientes infectados com SARS-CoV-2, internados em UTI sob VM, são mais predispostos à colonização por Candida spp., que pode ter um desfecho fatal nesses pacientes.(AU)

Justificación y objetivos: muchos pacientes con COVID-19 requieren hospitalización debido a complicaciones respiratorias graves y se someten a cuidados intensivos con soporte de ventilación mecánica (VM). Asociado a esta situación, hay un aumento de las coinfecciones fúngicas, lo que repercute negativamente en el desenlace de la COVID-19. En este sentido, este estudio pretendió comparar la incidencia de Candida spp. en el tracto respiratorio de pacientes ingresados en las Unidades de Cuidados Intensivos (UCI) COVID y General de un hospital escuela en 2021. Métodos: los resultados de muestras de aspirado traqueal protegidas de 556 pacientes ingresados en la UCI COVID y 260 en el UCI General, así como los respectivos registros. Resultados: de los pacientes analizados, 38 presentaron muestra positiva a Candida en UCI COVID y 10 en UCI General, con una incidencia de 68,3/1000 y 38,5/1000, respectivamente. Los machos predominaban en ambas alas. El grupo de edad más afectado fue la población mayor de 60 años, y la hospitalización promedio en la UCI COVID fue de 22,1 años, y en la UCI General, de 24,2. Conclusiones: Candida albicans fue la especie aislada con mayor frecuencia, y la tasa de mortalidad en pacientes positivos para Candida fue mayor en pacientes con COVID-19 en comparación con los pacientes en la UCI General, lo que sugiere que los pacientes infectados con SARS-CoV-2, ingresados en la UCI bajo VM, están más predispuestos a la colonización por Candida spp., lo que puede tener un desenlace fatal en estos pacientes.(AU)

Enzyme production by Candida albicans and Candida dubliniensis in periodontal HIV-positive patients receiving and not receiving antiretroviral therapy / Producción enzimática de Candida albicans y Candida dubliniensis de pacientes con periodontitis VIH+ con y sin tratamiento antirretroviral

ABSTRACT Candida dubliniensis (Cd) and Candida albicans (Ca) are the most frequently isolated yeasts in HIV+ patients. Some of the enzymes produced by these yeasts are considered virulence factors since they contribute to pathogenicity of Candida spp. The aim of the present study was to compare production of enzymes such as phospholipase (Ph), proteinase (P), and hemolysin (H) by Cd and Ca strains isolated from periodontal HIV-positive patients receiving and not receiving highly active antiretroviral therapy (HAART). Subgingival biofilm samples were obtained using paper points, and a sample of oral mucosa was taken using a swab. Phenotypic and molecular methods were used to isolate 39 strains of Candida, including 25 strains of Cd and 14 strains of Ca, obtained from 33 periodontal pocket samples and 6 oral mucosa samples collected from 15 HIV+ patients (8 receiving and 7 not receiving HAART). Malt egg-yolk agar, albumin agar and blood agar were used to evaluate pH, P and H production respectively. The strains were inoculated in duplicate and incubated at 37 ºC. Colony and halo diameters were measured. A greater proportion of Ca was observed in patients not receiving HAART, and a higher proportion of Cd was observed in those under HAART, Chi2 p< 0.001. Phospholipase production was observed in 92.9% percent of isolated Ca strains but in none of the isolated Cd strains. Proteinase production was high in Ca and Cd strains isolated from patients not receiving HAART. Hemolysin production was observed in all the studied strains, though it was significantly higher (p=0.04) in Ca and Cd strains isolated from patients not receiving HAART. To sum up, the proportion of Candida dubliniensis strains was highest in the subgingival biofilm of patients receiving HAART, and Cd strains were found to express fewer virulence factors than Ca strains.

RESUMEN Las levaduras más aisladas en pacientes VIH+ son Candida dubliniensis (Cd) y Candida albicans (Ca). Algunas de sus enzimas constituyen factores de virulencia ya que favorecen la diseminación tisular. El objetivo fue comparar la producción de enzimas como fosfolipasa (F), proteinasa (P) y hemolisina (H) en cepas de Cd y Ca aisladas de pacientes VIH+ tratados y no tratados con antirretrovirales (TARGA). Se realizó la toma del biofilm de placa subgingival con conos de papel y la muestra de la mucosa bucal con hisopo. Se aislaron y tipificaron por métodos fenotípicos y moleculares 39 cepas: 25 de Cd y 14 Ca, obtenidas 33 de bolsas periodontales y 6 de mucosa bucal de 15 pacientes VIH+ (8 con y 7 sin tratamiento). Se utilizó agar malta con yema de huevo, agar albúmina y agar sangre para demostrar la producción de F, P y H, respectivamente. Se inocularon por duplicado e incubaron a 37°C. Se midieron los diámetros de las colonias y los de hidrólisis alrededor de las mismas. Se observó mayor proporción de Ca en los pacientes sin tratamiento y mayor proporción de Cd en los con tratamiento; Chi2 p< 0.001. El 92,9% de las Ca estudiadas, fueron productoras de fosfolipasa. En tanto que ninguna Cd produjo la enzima. En cuanto a la producción de proteinasa se observa una alta producción tanto en las cepas de Ca, como en las Cd aisladas en los pacientes no tratados. Todas las cepas estudiadas produjeron hemolisina, observándose una diferencia estadísticamente significativa (p=0,04) en ambas especies a favor de la alta producción de la enzima en las cepas obtenidas de pacientes no tratados. Podemos concluir que en el biofilm subgingival, en los pacientes bajo TARGA, se aíslan mayor proporción de Candida dubliniensis las cuales expresan menos factores de virulencia.

Rastreio de leveduras patogênicas isoladas de fezes de pombos no Distrito Federal-Pombos: propagadores de fungos patogênicos? / Screening for pathogenic yeasts isolated from pigeon droppings in the Federal District-Pigeons: propagators of pathogenic fungi? / Detección de levaduras patógenas aisladas de excrementos de paloma en el Distrito Federal-Palomas: ¿propagadores de hongos patógenos?

Objetivo: Investigar a presença de fungos patogênicos isolados a partir de amostras de fezes de pombos, em locais de atenção a pacientes imunocomprometidos no DF. Método: As amostras foram coletadas em hospitais onde se oferta atendimento a pacientes HIV/AIDS e que tenha uma presença massiva de pombos. Colônias de leveduras foram previamente selecionados em meio Ágar Sabouraud Dextrose acrescido de cloranfenicol, seguindo-se com análise microscópica das estruturas leveduriformes. Colônias de leveduras com suspeita de pertencerem ao gênero Candida spp. ou Cryptococcus sp., foram inoculadas no meio Ágar Cromogênico para identificação das espécies de Candida e em meio Ágar quimicamente definido para indução dos fenótipos de virulência característicos de Cryptococcus sp. Resultados: 100% das amostras analisadas apresentaram crescimento de leveduras do gênero Candida spp. e Rhodotorula sp. No meio Ágar Cromogênico foram identificadas nas amostras C. krusei em 75%; C. tropicalis em 50% e C. glabrata em 15%. Em 15% das amostras foi identificado leveduras do gênero Cryptococcus sp. Conclusão: Dados deste estudo sugerem que fezes de pombo podem estar dispersando leveduras patogênicas e contribuindo com a incidência de infecções fúngicas no DF.

Objective: To investigate the presence of pathogenic fungi isolated from pigeon stool samples, in places of care for immunocompromised patients in the Federal District. Method: The samples were collected in hospitals where care is offered to HIV / AIDS patients and which has a massive presence of pigeons. Yeast colonies were previously selected on Sabouraud Dextrose Agar plus chloramphenicol, followed by microscopic analysis of the yeast structures. Yeast colonies suspected of belonging to the genus Candida spp. or Cryptococcus sp., were inoculated in the Chromogenic Agar medium to identify Candida species and in chemically defined Agar medium to induce the virulence phenotypes characteristic of Cryptococcus sp. Results: 100% of the analyzed samples showed growth of yeasts of the genus Candida spp. and Rhodotorula sp. In the chromogenic agar medium, 75% were identified in C. krusei samples; C. tropicalis in 50% and C. glabrata in 15%. In 15% of the samples, yeasts of the genus Cryptococcus sp. Conclusion: Data from this study suggest that pigeon feces may be dispersing pathogenic yeasts and contributing to the incidence of fungal infections in DF

Objetivo: Investigar la presencia de hongos patógenos aislados de muestras de heces de palomas, en los lugares de atención a pacientes inmunodeprimidos del Distrito Federal. Método: Las muestras se recolectaron en hospitales donde se brinda atención a pacientes con VIH / SIDA y que tiene una presencia masiva de palomas. Las colonias de levadura se seleccionaron previamente en Sabouraud Dextrose Agar más cloranfenicol, seguido de un análisis microscópico de las estructuras de la levadura. Las colonias de levaduras sospechosas de pertenecer al género Candida spp. o Cryptococcus sp., se inocularon en medio de agar cromogénico para identificar especies de Candida y en medio de agar químicamente definido para inducir los fenotipos de virulencia característicos de Cryptococcus sp. Resultados: el 100% de las muestras analizadas presentó crecimiento de levaduras del género Candida spp. y Rhodotorula sp. En el medio agar cromogénico, el 75% se identificó en muestras de C. krusei; C. tropicalis en 50% y C. glabrata en 15%. En el 15% de las muestras, levaduras del género Cryptococcus sp. Conclusión: Los datos de este estudio sugieren que las heces de las palomas pueden estar dispersando levaduras patógenas y contribuyendo a la incidencia de infecciones fúngicas en el DF.

Wide diversity of fungal species found in wellwater for human consumption: an analytical cross-sectional study

ABSTRACT BACKGROUND: Fungi are ubiquitous in the environment. They are able to grow in water and many of them may be opportunistic pathogens. OBJECTIVE: The aims were to identify fungi in registered wells (RWs) and nonregistered wells (NRWs) that tap into groundwater; and to correlate the results from physicochemical assays on this water (free residual chlorine and pH) with the presence of fungi. DATA AND SETTING: Analytical cross-sectional quantitative study on groundwater wells in São José do Rio Preto, São Paulo, Brazil. METHODS: 52 samples of 500 ml of water were collected from RWs and 107 from NRWs. These were sent to a microbiology laboratory to identify any fungi that were present. In addition, free residual chlorine and pH were measured immediately after sample collection. Several statistical analysis tests were used. RESULTS: Fungal contamination was present in 78.8% of the samples from RWs and 81.3% from NRWs. Filamentous fungi were more prevalent than yeast in both types of wells. There was no significant difference in presence of fungi according to whether chloride and pH were within recommended levels in RWs; or according to whether pH was within recommended levels in NRWs. Furthermore, there was no statistical difference in the levels of fungal contamination between RWs and NRWs. CONCLUSION: Both RWs and NRWs are potential reservoirs for many types of fungi. Many of these may become opportunistic pathogens if they infect immunosuppressed individuals. Furthermore, this study confirms that fungi are able to grow even when chlorine and pH parameters are within the standards recommended.

Candida auris infection in the central catheter of a patient without sepsis symptoms / Infección por Candida auris en el catéter central de un paciente sin síntomas de sepsis

Abstract Background: Candida auris is an emerging yeast frequently reported as resistant to multiple antifungal drugs commonly used to treat Candida infections. This specie can colonize the patient's skin and has great ability for producing outbreaks in hospitals. C. auris is phylogenetically related to other Candida species, can be misidentified using conventional biochemical or commercial methods and requires specific technology for its identification. Case report: We report the first isolate of C. auris in Cali, Colombia, from a central venous catheter in a 37-year-old patient with rheumatoid arthritis and endocarditis who did not have symptoms of sepsis. The yeast was initially misidentified as C. haemulonii using the Phoenix system and subsequently identified as C. auris by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The broth microdilution method was used to determine the minimum inhibitory concentration; the isolate was susceptible to fluconazole, itraconazole, voriconazole and amphotericin B. Conclusions: This report contributes to knowledge of the epidemiology of C. auris infections in individuals with underlying disease and describes an isolate with a behavior different from what is usually reported.

Resumen Antecedentes: Candida auris es una levadura emergente, informada con frecuencia como resistente a diversos antifúngicos usados comúnmente para tratar infecciones por Candida. Esta especie puede colonizar la piel y tiene gran capacidad de producir brotes en ambientes hospitalarios. Está filogenéticamente relacionada con otras especies de Candida, es mal identificada por los métodos bioquímicos o comerciales, y requiere tecnología específica para su identificación. Reporte de caso: Se informa el primer aislamiento de C. auris en Cali, Colombia en un paciente de 37 años con artritis reumatoide y endocarditis, sin síntomas de sepsis, a partir de la punta de catéter venoso central. La levadura inicialmente se identificó como C. haemulonii por el sistema Phoenix® y posteriormente como C. auris por espectrometría de masas desorción/ionización láser asistida por una matriz con detección de masas por tiempo de vuelo (MALDI-TOF MS). Se determinó la concentración inhibitoria mínima por el método de microdilución en caldo que mostró un aislamiento sensible a fluconazol, itraconazol, voriconazol y anfotericina B. Conclusión: Este informe contribuye al conocimiento de la epidemiología de las infecciones por C. auris en individuos con enfermedad subyacente y describe un aislamiento con un comportamiento diferente a lo indicado en otros estudios.

Osteomielitis fúngica en niños quemados / Fungal osteomyelitis in burned children

Introducción: En pacientes pediátricos quemados la osteomielitis fúngica es una complicación infrecuente que conduce a una significativa morbilidad. La información en la literatura está limitada a unos escasos reportes de casos. Objetivo: Describir las características clínicas, epidemiológicas y de evolución de niños quemados con osteomielitis fúngica. Métodos: Se llevo a cabo un estudio retrospectivo y descriptivo de pacientes mayores de 1 mes y menores de 18 años quemados con osteomielitis fúngica internados en el hospital Juan P. Garrahan, un hospital terciario en Buenos Aires, Argentina. Resultados: entre enero del 2007 y enero del 2017, de 600 niños quemados, 9 pacientes presentaron diagnóstico confirmado de osteomielitis fúngica. La mediana de edad fue de 42.5 meses (RIC, 27-118 meses) y la mediana de superficie quemada fue de 33.5% (RIC, 18.5-58%). La osteomielitis fue diagnosticada con una mediana de 30 días luego de la quemadura. Las localizaciones más frecuentes de osteomielitis fueron los miembros superiores y a nivel de calota. Los microorganismos aislados a partir del cultivo de hueso fueron: Fusarium spp. en tres pacientes, Mucor spp. en un paciente; Trichosporon asahii en un paciente; Cándida albicans en dos pacientes y Candida parapsilosis en dos pacientes. En dos casos la infección fúngica fue asociada con aislamientos bacteriano concomitante. Todos los pacientes presentaron hallazgos histopatológicos compatibles con osteomielitis. La mediana de tiempo de tratamiento fue de 44.5 días (RIC, 34.5- 65.5 días). Seis pacientes (67%) presentaron secuela motora. Conclusión: La osteomielitis fúngica fue infrecuente Candida spp. y Fusarium spp. fueron los hongos más comúnmente identificados. La secuela funcional fue frecuente (AU)

Introduction: In pediatric burn patients fungal osteomyelitis is a rare complication that leads to significant morbidity. Data in the literature are limited to sporadic case reports. Objective: To describe the clinical and epidemiological features and outcome in burned children with fungal osteomyelitis. Methods: A retrospective descriptive study was conducted in burn patients older than 1 month and younger than 18 years admitted to Hospital Juan P. Garrahan, a tertiary hospital in Buenos Aires, Argentina. Results: Between January 2007 and January 2017, of 600 burned children, nine had a confirmed diagnosis of fungal osteomyelitis. Median age was 42.5 months (IQR, 27-118 months) and median burn surface was 33.5% (IQR, 18.5-58%). Osteomyelitis was diagnosed at a median of 30 days after the burn. The most common location of osteomyelitis were the upper limbs and skull. The microorganisms isolated form bone cultures were Fusarium spp. in three patients, Mucor spp. in one patient; Trichosporon asahii in one patient; Candida albicans in two patients; and Candida parapsilosis in two patients. In two cases the funal infection was associated with concomitant bacterial isolation. In all patients, the histopathological findings were compatible with osteomyelitis. Median duration of treatment was 44.5 days (IQR, 34.5-65.5 days). Six patients (67%) had motor sequelae. Conclusion: Fungal osteomyelitis is a rare disease. Candida spp. and Fusarium spp. were most frequently identified fungi. Functional sequelae were common (AU)

MALDI-TOF mass spectrometry identification of yeast-form fungi: a comparison between methods

Introduction: Identification of yeast species has clinical and epidemiological value. Different methods can be used, such as chromogenic media, microculture on corn meal agar with Tween 80, as well as conventional biochemical and automated methods. Recently, proteomic studies employing matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry have been a major advance in diagnosis due to speed of execution and accuracy of results. Methods: For this study, 79 yeast samples were submitted to identification using chromogenic medium, microculture on corn meal-Tween 80 agar, VITEK 2 Compact identification, and MALDI-TOF mass spectrometry. Results: Most of the 79 samples were identified, with differences in the performance of the methods used. Colonial morphology and microscopy were compatible with the genus Candida. MALDI-TOF mass spectrometry had the best performance, with 78 strains identified (98.7%), compared to VITEK 2 Compact (92.4%) and microculture on corn meal agar (70.9%). Conclusions: MALDI-TOF mass spectrometry using the VITEK MS instrument performed best and has proven to be a revolutionary method in clinical microbiology laboratories. Regarding the identification of C. albicans and C. tropicalis, the chromogenic medium had excellent performance, thus being a good option to optimize the process. (AU)

Relationship between acetaldehyde concentration in mouth air and characteristics of microbiota of tongue dorsum in Japanese healthy adults: a cross-sectional study

Abstract Acetaldehyde, associated with consumption of alcoholic beverages, is known to be a carcinogen and to be related to the tongue dorsum. Objective The aim of this study was to investigate the relationship between acetaldehyde concentration in mouth air and bacterial characteristics on the tongue dorsum. Methodology Thirty-nine healthy volunteers participated in the study. Acetaldehyde concentrations in mouth air were evaluated by a high-sensitivity semiconductor gas sensor. A 16S rRNA gene sequencing technique was used to compare microbiomes between two groups, focusing on the six samples with the highest acetaldehyde concentrations (HG) and the six samples with lowest acetaldehyde concentrations (LG). Results Acetaldehyde concentration increased in correlation with the increase in bacterial count (p=0.048). The number of species observed in the oral microbiome of the HG was higher than that in the oral microbiome of the LG (p=0.011). The relative abundances of Gemella sanguinis, Veillonella parvula and Neisseria flavescens in the oral microbiome of the HG were higher than those in the oral microbiome of the LG (p<0.05). Conclusion Acetaldehyde concentration in mouth air was associated with bacterial count, diversity of microbiome, and relative abundance of G. sanguinis, V. parvula, and N. flavescens.

ERG11 gene polymorphisms and susceptibility to fluconazole in candida isolates from diabetic and kidney transplant patients

Abstract INTRODUCTION: Candidiasis is the most frequent opportunistic mycosis in humans and can cause mortality, particularly in immunodeficient patients. One major concern is the increasing number of infections caused by drug-resistant Candidas trains, as these cannot be efficiently treated with standard therapeutics. The most common mechanism of fluconazole resistance in Candida is mutation of ERG11, a gene involved in the biosynthesis of ergosterol, a compound essential for cell integrity and membrane function. METHODS: Based on this knowledge, we investigated polymorphisms in the ERG11 gene of 3 Candida species isolated from immunocompromised and immunocompetent patients. In addition, we correlated the genetic data with the fluconazole susceptibility profile of the Candida isolates. RESULTS: A total of 80 Candida albicans, 8 Candida tropicalis and 6 Candida glabrata isolates were obtained from the saliva of diabetic, kidney transplant and immunocompetent patients. Isolates were considered susceptible to fluconazole if the minimum inhibitory concentration was lower than 8 μg/mL. The amino acid mutations F105L, D116E, K119N, S137L, and K128T were observed in C. albicans isolates, and T224C and G263A were found in C. tropicalis isolates. CONCLUSIONS: Despite the high number of polymorphisms observed, the mutations occurred in regions that are not predicted to interfere with ergosterol synthesis, and therefore are not related to fluconazole resistance.

Presença de Candida spp. em usuários de próteses dentárias removíveis / Presencia de Candida spp. en usuarios de prótesis dentales removible / Presence of Candida spp. in patients who wear dentures

Introdução: O uso contínuo das próteses removíveis e a higiene inadequada das mesmas podem levar ao desenvolvimento da estomatite protética. Doença frequente em usuários de próteses e muito associada à Candida spp. Objetivo: determinar características sociais e os cuidados dos indivíduos com a higienização das próteses dentárias removíveis, assim como a presença de espécies de Candida em usuários de próteses numa comunidade do município de Cabedelo, Paraíba, Brasil. Métodos: Tratou-se de um estudo piloto, transversal, observacional e laboratorial com a avaliação de 14 voluntários. Foram utilizados formulários de investigação e realizada a coleta, o isolamento e identificação do material biológico. Resultados: Os participantes da pesquisa apresentaram renda e escolaridade baixas. A higiene diária das próteses, a escovação com dentifrício, a não remoção das próteses para dormir e a utilização das mesmas por mais de seis anos foram verificadas na maior parte da amostra. Conclusão: Demonstrou-se uma alta presença de Candida spp. nos usuários de próteses dentárias removíveis e Candida albicans foi a espécie mais prevalente(AU)

Introducción: El uso continuo de las prótesis removibles y la higiene inadecuada de estas pueden conducir al desarrollo de la estomatitis protésica, enfermedad frecuente en usuarios de prótesis y muy asociada a la Candida spp. Objetivo: determinar las características sociales y los cuidados de los individuos con la higienización de las prótesis dentales removibles, así como la presencia de especies de Candida en usuarios de prótesis en una comunidad del municipio de Cabedelo, Paraíba, Brasil. Métodos: Se trata de un estudio piloto, transversal, observacional y de laboratorio con la evaluación de 14 voluntarios. Se utilizaron formularios de investigación y se realizó la recolección, el aislamiento e identificación del material biológico. Resultados: Los participantes de la encuesta presentaron ingresos y escolaridad bajos. La higiene diaria de las prótesis, el cepillado con dentífrico, la no remoción de las prótesis para dormir y la utilización de estas durante más de seis años se verificaron en la mayor parte de la muestra. Conclusión: Se demostró una alta presencia de Candida spp. en los usuarios de prótesis dentales removibles y Candida albicans fue la especie que más predominó(AU)

Introduction : The continued use of dentures and inadequate hygiene may lead to the development of denture stomatitis, a frequent disease in patients who wear dentures and very much associated with Candida spp. Objective : To determine the social characteristics and the care of the individuals with the hygiene of their dentures, as well as the presence of Candida species in patients who wear dentures from a community of Cabedelo Municipality, Paraíba, Brazil. Methods : This is a pilot, cross-sectional, observational and laboratory study with the evaluation of 14 volunteers. Research forms were used and the collection, isolation and identification of the biological material were carried out. Results : The participants had low income and schooling. The daily hygiene of the prostheses, brushing with dentifrice, the non-removal of the prosthesis for sleeping and their use for more than six years were verified in most of the sample. Conclusions : A high presence of Candida spp. in the patients who wear dentures was shown, while C. albicans was the most prevalent species(AU)

Identification of Fungal Pathogens in Otomycosis and Their Drug Sensitivity: Our Experience

Abstract Introduction Otomycosis is a common problem in otolaryngology practice. However, we usually encounter some difficulties in its treatment because many patients show resistance to antifungal agents, and present high recurrence rate. Objectives To determine the fungal pathogens that cause otomycosis as well as their susceptibility to the commonly used antifungal agents. Additionally, to discover the main reasons for antifungal resistance. Methods We conducted an experimental descriptive study on 122 patients clinically diagnosed with otomycosis from April 2016 to April 2017. Aural discharge specimens were collected for direct microscopic examination and fungal culture. In vitro antifungal susceptibility testing was performed against the commonly used antifungal drugs. We tested the isolated fungi for their enzymatic activity. Results Positive fungal infection was found in 102 samples. The most common fungal pathogens were Aspergillus and Candida species, with Aspergillus niger being the predominant isolate (51%). The antifungal susceptibility testing showed that mold isolates had the highest sensitivity to voriconazole (93.48%), while the highest resistance was to fluconazole (100%). For yeast, the highest sensitivity was to nystatin (88.24%), followed by amphotericin B (82.35%), and the highest resistance was to terbinafine (100%), followed by Itraconazole (94.12%). Filamentous fungi expressed a high enzymatic ability, making them more virulent. Conclusion The Aspergillus and Candida species are the most common fungal isolates in otomycosis. Voriconazole and Nystatin are the medications of choice for the treatment of otomycosis in our community. The high virulence of fungal pathogens is owed to their high enzymatic activity. Empirical use of antifungals should be discouraged.

Comparison of the identification results of Candida species obtained by BD PhoenixT and Maldi-TOF (Bruker Microflex LT Biotyper 3.1) / Comparación de los resultados de identificación de especies del género Candida obtenidos por BD PhoenixT y Maldi-TOF (Bruker Microflex LT Biotyper 3.1)

In patients with invasive fungal infections, the accurate and rapid identification of the genus Candida is of utmost importance since antimycotic sensitivity is closely related to the species. The aim of the present study was to compare the identification results of species of the genus Candida obtained by BD PhoenixT (Becton Dickinson -#91;BD-#93;) and Maldi-TOF MS (Bruker Microflex LT Biotyper 3.1). A total of 192 isolates from the strain collection belonging to the Mycology Network of the Autonomous City of Buenos Aires, Argentina, were analyzed. The observed concordance was 95%. Only 10 strains (5%) were not correctly identified by the BD PhoenixT system. The average identification time with the Yeast ID panels was 8h 22 min. The BD PhoenixT system proved to be a simple, reliable and effective method for identifying the main species of the genus Candida.

En pacientes con infecciones fúngicas invasoras, la identificación certera y rápida de las especies del género Candida es de suma importancia, ya que la sensibilidad a los antifúngicos está íntimamente relacionada con la especie. El objetivo del presente estudio fue comparar los resultados de identificación de especies del género Candida obtenidos con el equipo comercial BD PhoenixT (Becton Dickinson -#91;BD-#93;) y con la técnica de Maldi-TOF MS (Bruker Microflex LT Biotyper 3.1.) Se analizaron 192 aislamientos provenientes del cepario perteneciente a la Red deMicología de la Ciudad Autónoma de Buenos Aires, Argentina. La concordancia observada fue del 95%. Solo 10 cepas (5%) no fueron identificadas correctamente por el sistema BD PhoenixT. El tiempo promedio de identificación con los paneles Yeast ID fue de 8 h 22 min. El sistema BD PhoenixT demostró ser un método simple, confiable y efectivo para la identificación de las principales especies del género Candida.

Evaluation of the frequency of Candida spp. in hospitalized and non-hospitalized subjects / Avaliação da frequência de candida spp. em indivíduos hospitalizados e não hospitalizados

Abstract The aim of this study was to evaluate the frequency of Candida species between a non-hospitalized and a hospitalized population. For this purpose, samples of saliva were sampled through sterile swabs, moistened in peptone water and rubbed in the oral cavity of 140 individuals, from which, 70 were hospitalized patients from the Medical Clinic of a Teaching Hospital and the other 70 were non-hospitalized subjects. All saliva samples were plated in Sabouraud Dextrose agar added with Chloramphenicol and incubated at 36 °C for 48 hours. The morphology identification was performed through macroscopic and microscopic characterization, the CHROMagar Candida medium and the VITEK® system Yeast Biochemical Card (bio Mérieux SA, France). The results showed a colonization of Candida spp. in 85.7% the hospitalized individuals, where the species found were C. albicans (60%), C. tropicalis (23.4%), C. krusei (3.3%) and Candida spp. (13.3%). In the non-hospitalized individuals the colonization by Candida spp was 47.1%, and the species found were: C. albicans (45.5%), C.krusei (9.1%), C. guilliermondii (9.1% %), C. tropicalis (3.0%), C. famata (3.0%) and Candida spp. (30.3%). In spite of their presence in oral cavity in both groups, Candida spp. was more frequently isolated in hospitalized individuals, who were 6.73 times more likely to have this fungus in the oral cavity and were 3.88 times more likely to have Candida albicans.

Resumo O objetivo deste estudo foi avaliar a frequência de espécies de Candida entre uma população de indivíduos não-hospitalizados e hospitalizados. Para isto, amostras de saliva foram coletadas através de swabs estéreis, umedecidas em água de peptona e friccionadas na cavidade bucal de 140 indivíduos, dos quais 70 eram pacientes internados em uma Clínica Médica de um Hospital Escola e os outros 70 eram indivíduos não hospitalizados sem contato com ambiente hospitalar. Todas as amostras de saliva foram plaqueadas em ágar Sabouraud dextrose adicionadas de cloranfenicol e incubadas a 36 °C durante 48 horas. A identificação morfológica foi realizada através da caracterização macroscópica e microscópica, com o meio CHROMagar Candida e do sistema VITEK® Biochemical Card (bio Mérieux SA, França). Os resultados mostraram uma colonização de Candida spp. em 85,7% dos indivíduos hospitalizados, onde as espécies encontradas foram: C.albicans (60%), C. tropicalis (23,4%), C. krusei (3,3%) e Candida spp. (13,3%). Nos indivíduos não-hospitalizados a colonização por Candida spp foi de 47,1%, e as espécies encontradas foram: C. albicans (45,5%), C. krusei (9,1%), C. guilliermondii (9,1%), C. tropicalis (3,0%), C. famata (3,0%) e Candida spp. (30,3%). Apesar de sua presença na cavidade oral em ambos os grupos, Candida spp. foi mais freqüentemente isolada em indivíduos hospitalizados, que foram 6,73 vezes mais propensos a ter este fungo na cavidade oral e foram 3,88 vezes mais propensos a ter Candida albicans.

Tratamiento satisfactorio con fluconazol de la candidemia relacionada con el catéter porCandida haemulonii: informe sobre dos pacientes neonatales / Successful fluconazole treatment of catheter-related candidemia caused by Candida haemulonii: report of 2 neonatal cases

La Candida haemulonii forma parte de la especie Candida no albicans. La candidemia por C. haemulonii es sumamente infrecuente, pero mortal, en los recién nacidos. Se informa sobre los dos primeros recién nacidos con candidemia por C. haemulonii en China tratados con fluconazol y se revisan dos artículos informados con anterioridad. Nuestro informe incrementa la sensibilización sobre la candidemia por C. haemulonii en recién nacidos críticos y resalta la importancia de un diagnóstico y un tratamiento tempranos de esta infección mortal.

Candida haemulonii forms part of the non-albicans Candida species. The candidemia caused by C. haemulonii is extremely rare but fatal in neonates. We reported the first two neonates with C. haemulonii candidemia in China which were treated with fluconazole and reviewed two papers previously reported. Our report adds further awareness on C. haemulonii candidemia in critical neonates and points out the importance of an early diagnosis and treatment of this fatal infection.

Concordance analysis between different methodologies used for identification of oral isolates of Candida species / Análisis de concordancia de diferentes metodologías para la identificación de aislamientos orales de especies de Candida

Abstract Background: The yeasts species determination is fundamental not only for an accurate diagnosis but also for establishing a suitable patient treatment. We performed a concordance study of five methodologies for the species identification of oral isolates of Candida in Colombia. Methods: Sixty-seven Candida isolates were tested by; API® 20C-AUX, Vitek®2 Compact, Vitek®MS, Microflex® and a molecular test (panfungal PCR and sequencing). The commercial cost and processing time of the samples was done by graphical analysis. Results: Panfungal PCR differentiated 12 species of Candida, Vitek®MS and Microflex® methods identified 9 species, and API® 20C-AUX and Vitek®2 Compact methods identified 8 species each. Weighted Kappa (wK) showed a high agreement between Panfungal PCR, Vitek®MS, Microflex® and API® 20C-AUX (wK 0.62-0.93). The wK that involved the Vitek®2 Compact method presented moderate or good concordances compared with the other methods (wK 0.56-0.73). Methodologies based on MALDI TOF MS required 4 minutes to generate results and the Microflex® method had the lowest selling price. Conclusion: The methods evaluated showed high concordance in their results, being higher for the molecular methods and the methodologies based on MALDI TOF. The latter are faster and cheaper, presenting as promising alternatives for the routine identification of yeast species of the genus Candida.

Resumen Introducción: La clasificación a nivel de especies de las levaduras del género Candida de origen clínico es fundamental para el diagnóstico y la instauración de un adecuado tratamiento para el paciente. Se realizó un estudio de concordancia de cinco metodologías usadas para la identificación de aislamientos orales de Candida spp en Colombia. Métodos: Sesenta y siete aislamientos de Candida spp fueron identificados a nivel de especie utilizando; API® 20 C AUX‚ Vitek® 2 Compact, MALDI TOF (Vitek® MS y Microflex®) y una prueba molecular, PCR Panfungal y secuenciación. Un análisis del costo comercial y tiempo de procesamiento de las muestras por cada método fue realizado mediante el análisis gráfico de ambas variables. Resultados: La PCR Panfungal y secuenciación diferenció 12 especies de Candida‚ los métodos Vitek® MS y Microflex® identificaron 9 especies y los métodos API® 20 C AUX y Vitek® 2 Compact identificaron 8 especies. El análisis de Kappa ponderado (wK) demostró una concordancia alta entre los métodos PCR Panfungal y secuenciación‚ Vitek® MS‚ Microflex® y API® 20 C AUX‚ concordancias agrupadas en las categorías buena y muy buena (wK 0.62 - 0.93); los Kp que involucraron el método Vitek® 2 Compact presentaron concordancias moderadas o buenas frente a los otros métodos (wK 0.56 - 0.73). Las metodologías basadas en MALDI TOF MS requirieron 4 minutos para generar un resultado y el método Microflex® fue el método que en nuestro medio presentó el menor precio de venta del servicio. Conclusión: Los métodos evaluados presentaron una alta concordancia en sus resultados‚ siendo más alta para los métodos moleculares y las metodologías basadas en MALDI TOF MS; estas últimas son metodologías más rápidas, económicas y precisas, las cuales se presentan como alternativas prometedoras para la identificación rutinaria de especies de levaduras del género Candida.

Secular trends of candidemia at a Brazilian tertiary care teaching hospital

ABSTRACT Background Candidemia is the most frequent invasive fungal disease in hospitalized patients, and is associated with high mortality rates. The main objective of this study was to evaluate changes in the epidemiology of candidemia at a tertiary care hospital in a 21-year period. Methods We evaluated all episodes of candidemia diagnosed between 1996 and 2016 at a University-affiliated tertiary care hospital in Brazil. We arbitrarily divided the study period in 3: 1996-2002 (period 1), 2003-2009 (period 2) and 2010-2016 (period 3). Incidence rates were calculated using hospital admissions as denominator. Results We observed 331 episodes of candidemia. The incidence was 1.30 episodes per 1000 admissions, with no significant change over time. Candida albicans (37.5%), C. tropicalis (28.1%), C. parapsilosis (18.4%) and C. glabrata (6.9%) were the most frequent species. The proportion of patients receiving treatment increased (65.5%, 79.4% and 74.7% in periods 1, 2 and 3, respectively, p= 0.04), and the median time from candidemia to treatment initiation decreased from 4 days in period 1 (range 0-32 days) to 2 days in period 2 (range 0-33 days) and 2 days in period 3 (range 0-14 days, p< 0.001). We observed a significant decrease in the use of deoxycholate amphotericin B (47.4%, 14.8% and 11.9%), and an increase in the use of echinocandins (0%, 2.8% and 49.1%; p< 0.001). The APACHE II score increased over time (median 16, 17.5, and 22, p< 0.001). The overall 30-day mortality was 58.9%, and did not change significantly over the study period. Conclusions There was an improvement in patient care, with an increase in the proportion of patients receiving treatment and a decrease in the time to treatment initiation, but no improvement in the outcome, possibly because the proportion of sicker patients increased over time.

In vitro antifungal susceptibility of Candida speciesisolated from diabetic patients

Abstract INTRODUCTION This study aims to evaluate the antifungal susceptibility of different species of Candida isolated from diabetic patients against eight antifungal agents. METHODS Susceptibility testing of 111 clinical isolates of Candida species was performed against 8 antifungals using the M27-A3 protocol of the Clinical and Laboratory Standards Institute (CLSI). RESULTS Voriconazole, lanoconazole, and caspofungin showed the highest in vitro activity against all the isolates of C. albicans. Resistance against the tested antifungals was only observed in the C. albicans isolates. CONCLUSIONS Our finding revealed that resistance against amphotericin B, itraconazole, ketoconazole, posaconazole, and fluconazole can be observed in C. albicans.

The potential of the newly isolated thermotolerant yeast Pichia kudriavzevii RZ8-1 for high-temperature ethanol production

Abstract High potential, thermotolerant, ethanol-producing yeasts were successfully isolated in this study. Based on molecular identification and phylogenetic analysis, the isolated thermotolerant yeasts were clustered in the genera of Pichia kudriavzevii, Candida tropicalis, Candida orthopsilosis, Candida glabrata and Kodamea ohmeri. A comparative study of ethanol production using 160 g/L glucose as a substrate revealed several yeast strains that could produce high ethanol concentrations at high temperatures. When sugarcane bagasse (SCB) hydrolysate containing 85 g/L glucose was used as a substrate, the yeast strain designated P. kudriavzevii RZ8-1 exhibited the highest ethanol concentrations of 35.51 g/L and 33.84 g/L at 37 °C and 40 °C, respectively. It also exhibited multi-stress tolerance, such as heat, ethanol and acetic acid tolerance. During ethanol fermentation at high temperature (42 °C), genes encoding heat shock proteins (ssq1 and hsp90), alcohol dehydrogenases (adh1, adh2, adh3 and adh4) and glyceraldehyde-3-phosphate dehydrogenase (tdh2) were up-regulated, suggesting that these genes might play a crucial role in the thermotolerance ability of P. kudriavzevii RZ8-1 under heat stress. These findings suggest that the growth and ethanol fermentation activities of this organism under heat stress were restricted to the expression of genes involved not only in heat shock response but also in the ethanol production pathway.

Clinical and laboratorial features of oral candidiasis in HIV-positive patients

Abstract INTRODUCTION We describe the clinical and laboratorial features of oral candidiasis in 66 HIV-positive patients. METHODS: Polymerase chain reaction-based techniques were performed for differentiation of Candida spp. isolated from patients at a public teaching hospital in Midwest Brazil. RESULTS: Oral lesions, mainly pseudomembranous, were significantly related to higher levels of immunosuppression. Of 45 Candida isolates, 66.7% were C. albicans. Most of the isolates were susceptible to the antifungal drugs tested. CONCLUSIONS: Oral lesions were associated with higher immunosuppression levels. Lower susceptibility to antifungals by non-albicans isolates supports the importance of surveillance studies using susceptibility tests to aid in the treatment.