Expression of ERG11, ERG3, MDR1 and CDR1 genes in Candida tropicalis

Introduction. Drug resistance to azoles is a growing problem in the Candida genus. Objective. To analyze molecularly the genes responsible for fluconazole resistance in Candida tropicalis strains. Materials and methods. Nineteen strains, with and without exposure to fluconazole, were selected for this study. The expression of MDR1, CDR1, ERG11, and ERG3 genes was analyzed in sensitive, dose-dependent sensitive, and resistant strains exposed to different concentrations of the antifungal drug. Results. MDR1, ERG11 and ERG3 genes were significantly overexpressed in the different sensitivity groups. CDR1 gene expression was not statistically significant among the studied groups. Seven of the eight fluconazole-resistant strains showed overexpression of one or more of the analyzed genes. In some dose-dependent sensitive strains, we found overexpression of CDR1, ERG11, and ERG3. Conclusion. The frequency of overexpression of ERG11 and ERG3 genes indicates that they are related to resistance. However, the finding of dose-dependent resistant/sensitive strains without overexpression of these genes suggests that they are not exclusive to this phenomenon. More basic research is needed to study other potentially involved genes in the resistance mechanism to fluconazole.

Introducción. La farmacorresistencia a los azoles es un problema creciente en el género Candida. Objetivo. Analizar molecularmente los genes responsables de la resistencia a fluconazol en cepas de Candida tropicalis. Materiales y métodos. Para este estudio, se seleccionaron 19 cepas, con exposición a fluconazol y sin ella. Se analizó la expresión de los genes MDR1, CDR1, ERG11 y ERG3 en cepas sensibles, sensibles dependiente de la dosis, y resistentes, previamente expuestas a diferentes concentraciones del fármaco antifúngico. Resultados. Se encontró que los genes MDR1, ERG11 y ERG3 estaban significativamente sobreexpresados en los diferentes grupos de sensibilidad. La expresión del gen CDR1 no fue estadísticamente significativa entre los grupos estudiados. Siete de las ocho cepas resistentes a fluconazol mostraron sobreexpresión de uno o más de los genes analizados. En algunas cepas sensibles dependientes de la dosis, se encontró sobreexpresión de CDR1, ERG11 y ERG3. Conclusión. La sobreexpresión de los genes ERG11 y ERG3 indica que están relacionados con la resistencia de las cepas de Candida. Sin embargo, el hallazgo de cepas resistentes o sensibles según la dosis, sin sobreexpresión de estos genes, sugiere que pueden existir otros genes involucrados en este fenómeno. Se necesitan más investigaciones básicas que contribuyan al estudio de otros genes potencialmente involucrados en el mecanismo de resistencia al fluconazol.

Comportamiento de Candida en niños hospitalizados en unidades de cuidados intensivos / Incidence of Candida in children hospitalized in intensive care units

Introducción: La candidemia es considerada el tipo de fungemia más frecuente asociada a pacientes en unidades de cuidados intensivos. Es una afección rápidamente progresiva con elevada mortalidad, la cual presenta dificultad en un diagnóstico precoz y capacidad de resistencia al tratamiento. Objetivos: Caracterizar aspectos epidemiológicos, clínicos y microbiológicos de las especies de Candida aisladas en niños hospitalizados en unidades de cuidados intensivos. Métodos: Se realizó un estudio descriptivo y transversal en 143 aislamientos de especies de Candida obtenidas en hemocultivos de pacientes menores de 18 años, en el laboratorio de Microbiología del Hospital Universitario Ginecobstétrico «Mariana Grajales», en el período de enero de 2009 a diciembre de 2018. Se utilizaron tablas de contingencia (prueba de Ji Cuadrado y V de Cramer); se utilizaron las variables: edad, sexo, factores de riesgo, especies y servicios hospitalarios. Resultados: Los mayores aislamientos de candidemia correspondieron a pacientes del sexo femenino de edades comprendidas entre 29 días y 18 años. Todos fueron sometidos al uso de antimicrobianos y con ventilación mecánica. El mayor número de aislamientos en pacientes de 9 años correspondió a Candida tropicalis, que fue la más aislada en la Unidad de Neonatología y Cuidados Intensivos Pediátricos, no así en Cuidados Intermedios donde predominó Candida sin precisar especie. Por estas razones se puede señalar que existe una relación entre las especies y los servicios hospitalarios. Conclusiones: Predominaron pacientes del sexo femenino mayores de 29 días y hasta 18 años hospitalizados en la Unidad de Cuidados Intensivos Pediátricos; la especie más aislada fue Candida tropicalis.

Introduction: candidemia is considered the most frequent type of fungemia associated with patients hospitalized in intensive care units. It is a rapidly progressive condition with high mortality, difficult to diagnose early and resistant to treatment. Objective: to characterize the epidemiological, clinical and microbiological aspects of Candida species isolated from children hospitalized in intensive care units. Methods: a descriptive and cross-sectional study was carried out on 143 isolates of Candida species corresponding to blood cultures from patients under 18 years of age and obtained in the Microbiology laboratory at "Mariana Grajales" Gynecological and Obstetric University Hospital, from January 2009 to December 2018. Contingency tables were used (Chi- square and Cramer's V tests); age, gender, risk factors, species and hospital services were the used variables. Results: the largest candidemia isolates corresponded to female patients aged between 29 days and 18 years. All were subjected to the use of antimicrobials and mechanical ventilation. The highest number of isolates in 9-year-old patients corresponded to Candida tropicalis, which was the most isolated species in the Neonatology and Pediatric Intensive Care Unit, but not in the Intermediate Care Unit where Candida predominated without specifying the species. For these reasons, it can be pointed out that there is a relationship between species and hospital services. Conclusions: female patients older than 29 days and up to 18 years hospitalized in the Pediatric Intensive Care Unit predominated; the most isolated species was Candida tropicalis.

Candida Tropicalis Spondylodiscits in an Immunocompetent Host: A Case Report and Literature Review

Background Fungal spondylodiscitis is not common but should be suspected in some cases. Candida tropicalis infections are being more frequently diagnosed due to some factors related to the microorganism. Case Description A C. tropicalis spondylodiscitis is described in a 72-year-old man who was treated with a combination of echinocandin (micafungin) and surgery. Conclusion The presence of some risk factors should promptly raise the suspicion of fungal spondylodiscitis. Treatment should be instituted as early as possible for the best outcome for the patient.

Atividade antifúngica de isolados clínicos de Candida não-albicans aos óleos essenciais de Syzygium aromaticum (cravo-da-índia) e Eucalyptus globulus (eucalipto-comum) / Antifungal activity of clinical isolates of Candida non-albicans to the essential oils of Syzygium aromaticum (cloud) and Eucalyptus globulus (eucalyptus)

RESUMO: Introdução: O gênero Candida possui alta taxa de incidência no ser humano, sendo a espécie Candida albicans a mais isolada em infecções invasivas e superficiais. Porém, tem sido relatado um aumento considerável de espécies de Candidanão-albicans em infecções fúngicas. Os óleos essenciais, por serem voláteis, podem agir como sinais de comunicação química e arma de defesa. Objetivo: Avaliar a eficácia, in vitro, dos óleos essenciais de Syzygium aromaticum e Eu-calyptus globulus na inibição do crescimento de espécies de Candida não-albicans. Métodos: Para avaliação da atividade antifúngica de S. aromaticum e de E. globulus e do efeito dos seus óleos essenciais sobre a micromorfologia das espécies Candida krusei, Candida parapsilosis e Candida glabrata, foram empregadas, nesta ordem, a técnica de difusão em ágar e de microcultivo para leveduras. Resultados: Na técnica de difusão, o óleo essencial de S. aromaticum apresentou for-mação de halo de inibição para Candida krusei, Candida parapsilosis e Candida glabrata. O óleo de E. globulus, por sua vez, não apresentou crescimento de halos de inibição em nenhuma das concentrações testadas frente às três espécies de Candida não-albicans. Todavia, com o microcultivo, ambos os óleos essenciais se provaram, in vitro, eficazes antimi-crobianos tendo apresentado estruturas indicativas de atividade antifúngica na maior concentração dos óleos e diferentes graus de destruição celular nas demais concentrações. Conclusão: Nas condições deste estudo, concluiu-se que os pro-dutos avaliados exerceram atividade antifúngica contra cepas de Candida não-albicans, destacando-se o óleo essencial de S. aromaticum que apresentou atividade antimicrobiana em ambas as metodologias. (AU)

ABSTRACT: Introduction: The genus Candida possesses a high incidence of infection in humans, with the Candida albicans species being the most isolated in invasive and superficial infections. However, a considerable increase in non-albicans Candidaspecies has been reported in fungal infections. Essential oils, for being volatile, can act as chemical communication sig-nals and defense mechanisms. Objective: Evaluate the effectiveness, in vitro, of the essential oils of Syzygium aromati-cum and Eucalyptus globulus in inhibiting the growth of non-albicans Candida species. Methods: To evaluate the antifun-gal activity of S. aromaticum and E. globulus and the effect of their essential oils on the micromorphology of the species Candida krusei, Candida parapsilosis and Candida glabrata, in this order, the agar diffusion technique and microculture for yeasts. Results: In the diffusion technique, the essential oil of S. aromaticum presented an inhibition halo for Candida krusei, Candida parapsilosis, and Candida glabrata. E. globulus oil, on the other hand, did not show inhibition halo growth in any of the concentrations tested against the three non-albicans Candida species. However, in the microculture, both essential oils have proven to be effective antimicrobials having shown structures indicative of antifungal activity in the highest concentration and different degrees of cell destruction in the other concentrations. Conclusion: In the conditions of this study, it was concluded that the products evaluated exerted antifungal activity against non-albicans Candidastrains, highlighting the essential oil of S. aromaticum that showed antimicrobial activity in both methodologies. (AU)

Idiopathic ileal volvulus with multiple concomitant infections in a starving man

@#Background: Small bowel volvulus is defined as the torsion of the small intestine, potentially leading to bowel obstruction, gangrene and perforation. It is a rare condition, especially in adults. Case presentation: A 30-year-old man was retrieved from the jungle with severe weight loss and abdominal symptoms. He succumbed to death despite 22 days of intensive medical treatment. An autopsy revealed a ruptured gangrenous ileal volvulus with peritonitis and subdiaphragmatic abscess. Further laboratory analysis detected systemic Candida tropicalis and intestinal gramnegative bacterial sepsis, systemic Zika virus viremia, leptospirosis complicating rhabdomyolysis and disseminated intravascular coagulopathy, Type I Herpes Simplex virus infection of the tongue and upper gastrointestinal tract. The cause of death was the ruptured ileal volvulus, complicated with upper gastrointestinal bleeding due to Herpes simplex virus esophagitis in a malnourished patient with resolving leptospirosis and underlying Zika virus co-infection. Conclusion: Rare clinical scenarios of adult-onset intestinal volvulus with concomitant multiple infections precludes clinical diagnosis and early treatment, leading to devastating consequences of clinical outcome. The positive clinical and postmortem correlation is a good learning lesson in many disciplines of medicine and science.

Frecuencia y susceptibilidad antifúngica de Candida spp. (no C. albicans ) aislada de pacientes de unidades de cuidados críticos de un hospital de tercer nivel del norte del Perú / Frequency and antifungal susceptibility of Candida spp. (other than C. albicans ) isolated from patients admitted to critical care units of a third-level hospital in northern Peru

RESUMEN Objetivo Determinar la frecuencia y susceptibilidad antifúngica de Candida spp. (no C. albicans) aisladas de muestras de pacientes en cuidados críticos de un hospital de tercer nivel del norte del Perú. Materiales y métodos Se realizó un estudio transversal entre febrero del 2018 y mayo del 2019 en 232 muestras de pacientes hospitalizados en unidades de cuidados críticos del Hospital Regional Lambayeque (HRL). Candida ssp. (no C. albicans) fue identificada mediante las pruebas fenotípicas. La susceptibilidad antifúngica al fluconazol y voriconazol se determinó según la interpretación de los puntos de corte del documento M44-A2 del CLSI (por sus siglas en inglés, Clinical and Laboratory Standards Institute). Resultados Se estudiaron 232 muestras. La frecuencia de Candida spp. (no C. albicans) fue de 19,82 % (46) y C. albicans de 11,20 % (26). Las especies de Candida spp. (no C. albicans) fueron las siguientes: Treintaiuna de C. tropicalis (67,39 %), ocho de C. glabrata (17,39 %), cinco de C. parapsilosis (10,86 %) y dos de C. krusei (4,34 %). Los pacientes en quienes se aisló Candida spp. (no C. albicans) se caracterizaron por ser, en su mayoría, mujeres (56,52 %), que previamente habían usado antimicrobianos (67,39 %), y de 60 años a más (36,95 %). La muestra en la que se aisló en microorganismo con mayor frecuencia fue la orina (45,65 %). Respecto a la susceptibilidad antifúngica de las especies de Candida spp. (no C. albicans), encontramos resistencia a fluconazol (19,56 %) y a voriconazol (21,73 %). Conclusiones La alta frecuencia y predominio de especies de Candida spp. (no C. albicans) en pacientes de unidades de cuidados críticos de un hospital de tercer nivel en Chiclayo, y la resistencia que muestran a la acción de fluconazol y voriconazol son características de gran importancia en la morbilidad y mortalidad en estos pacientes.

ABSTRACT Objective To determine the frequency and antifungal susceptibility of Candida spp. (other than C. albicans) isolated from samples of patients admitted to critical care units of a third-level hospital in northern Peru. Materials and methods A cross-sectional study was conducted between February 2018 and May 2019 in 232 samples of patients admitted to critical care units of the Hospital Regional Lambayeque (HRL). Candida spp. (other than C. albicans) were identified using phenotypic tests. The antifungal susceptibility to fluconazole and voriconazole was determined according to the epidemiological cut-off values of CLSI document M44-A2. Results Two hundred thirty-two (232) samples were analyzed. The frequency of Candida spp. (other than C. albicans) was 19.82 % (46/232) and C. albicans was 11.20 % (26/232). Candida spp. (other than C. albicans) were the following: C. tropicalis (67.39 %, 31/46), C. glabrata (17.39 %, 8/46), C. parapsilosis (10.86 %, 5/46) and C. krusei (4.34 %, 2/46). Patients from whom Candida spp. (other than C. albicans) were isolated were mostly females (56.52 %), had a history of antimicrobial drugs consumption (67.39 %) and were 60 years of age and older (36.95 %). Most microorganisms were isolated from urine samples (45.65 %). Regarding the antifungal susceptibility of Candida spp. (other than C. albicans), 19.56 % and 21.73 % were resistant to fluconazole and voriconazole, respectively. Conclusions The high incidence and prevalence of Candida spp. (other than C. albicans) in patients admitted to critical care units of a third-level hospital in Chiclayo, together with resistance to fluconazole and voriconazole, are significant in the morbidity and mortality of these patients.

Nutrient compositions of distillers dried grain from rice husks with co-culture fermentation of Saccharomyces cerevisiae with Candida tropicalis

Aims@#Distillers dried grains are the nutrient rich co-product of dry-milled ethanol production. The present study aimed to prove that the nutritional composition of distillers dried grain from a crude hydrolysate of rice husk fermented by co-cultures of Saccharomyces cerevisiae with Candida tropicalis difference from unfermented crude rice husk hydrolysate and mono-cultured S. cerevisiae or C. tropicalis. @*Methodology and results@#The effects of mono- and co-cultures S. cerevisiae with C. tropicalis on the nutrient compositions of distillers dried grain were investigated. The crude rice husk hydrolysate in distilled water contained molasses, urea, sodium nitrate, ammonium nitrate, potassium phosphate and magnesium sulfate heptahydrate were fermented by mono- and co-cultures of S. cerevisiae with C. tropicalis for 7 days at 28-30 °C and stored with a relative humidity of 60-70% in the dark. A mono- and a co-culture fermentation of S. cerevisiae and C. tropicalis increased the crude protein, crude fat, crude fibres, ash, and calcium contents of the rice husk feedstock and decreased the metabolic energy reducing sugars. @*Conclusion, significance and impact of study@#Some nutrient components of the DDG crude rice husk hydrolysate performed higher than the non-fermentation of rice husks. The finding of this study will serve as a basic reference for future studies to utilize by-product of ethanol production from rice husks for animal feed formulation.

HPLC-DAD analysis and antifungal effect of Hyptis martiusii Benth (Lamiaceae) against Candida strains

Objective: To evaluate the anti Candida activity of Hyptis martiusii decoction and its major compound, caffeic acid alone or in the presence of fluconazole, as well as their cytotoxic effect. Methods: The decoction was characterized using high performance liquid chromatography coupled with diode array detector. For the antifungal activity, the minimum inhibitory concentration (MIC) and the potential effect of the decoction with the fluconazole were evaluated by microdilution method using 96-well microtiter trays. The osmotic fragility test was performed using erythrocytes under saline stress. All tests were performed in triplicate. Results: The chemical characterization of the decoction was performed by high performance liquid chromatography and revealed the presence of seven compounds, including caffeic acid as major constituent. The antifungal tests demonstrated that both decoction (DHm) and caffeic acid obtained from Hyptis martiusii presented MIC and MFC ≥ 4096 μg/mL against Candidaalbicans and Candida tropicalis strains. However, in the presence of fluconazole, DHm and caffeic acid presented IC50 of 2.60 and 2.53 μg/mL respectively, demonstrating significant synergistic effects against Candida strains. The modulator activity of DHm might be due to the presence of caffeic acid. Moreover, DHm and caffeic acid did not cause significant hemolytic effects, indicating that they present low cytotoxicity. Conclusions: These data indicate that DHm potentiates the activity of the fluconazole, without enhancement of the toxicity, encouraging further toxicological, pharmacological and phytochemical studies to provide consistent evidence of the potential of this plant to be used in drug development.

HPLC-DAD analysis and antifungal effect of Hyptis martiusii Benth (Lamiaceae) against Candida strains

Objective: To evaluate the anti Candida activity of Hyptis martiusii decoction and its major compound, caffeic acid alone or in the presence of fluconazole, as well as their cytotoxic effect. Methods: The decoction was characterized using high performance liquid chromatography coupled with diode array detector. For the antifungal activity, the minimum inhibitory concentration (MIC) and the potential effect of the decoction with the fluconazole were evaluated by microdilution method using 96-well microtiter trays. The osmotic fragility test was performed using erythrocytes under saline stress. All tests were performed in triplicate. Results: The chemical characterization of the decoction was performed by high performance liquid chromatography and revealed the presence of seven compounds, including caffeic acid as major constituent. The antifungal tests demonstrated that both decoction (DHm) and caffeic acid obtained from Hyptis martiusii presented MIC and MFC ≥ 4096 μg/mL against Candida albicans and Candida tropicalis strains. However, in the presence of fluconazole, DHm and caffeic acid presented IC

Pseudomembranous candidiasis by candida tropicalis in an immunocompromised patient: case report

Pseudomembranous candidiasis is the most frequent type of infection by candida spp., and candida albicans is the most common species to cause it. candidiasis can be due to other candida species less frequently, as is the case of candida tropicalis a pathogenic species that can cause infection in immunocompromised patients. the aim of this case report is to describe a pathological condition produce by candida tropicalis.

Interactions between Terpinen-4-ol and Nystatin on biofilm of Candida albicans and Candida tropicalis

Abstract The aim of this study was to evaluate the antifungal activity of Terpinen-4-ol associated with nystatin, on single and mixed species biofilms formed by Candida albicans and Candida tropicalis, as well as the effect of terpinen-4-ol on adhesion in oral cells and the enzymatic activity. The minimum inhibitory concentrations and minimum fungicide concentrations of terpinen-4-ol and nystatin on Candida albicans and Candida tropicalis were determined using the microdilution broth method, along with their synergistic activity ("checkerboard" method). Single and mixed species biofilms were prepared using the static microtiter plate model and quantified by colony forming units (CFU/mL). The effect of Terpinen-4-ol in adhesion of Candida albicans and Candida tropicalis in coculture with oral keratinocytes (NOK Si) was evaluated, as well as the enzymatic activity by measuring the size of the precipitation zone, after the growth agar to phospholipase, protease and hemolysin. Terpinen-4-ol (4.53 mg mL-1) and nystatin (0.008 mg mL-1) were able to inhibit biofilms growth, and a synergistic antifungal effect was showed with the drug association, reducing the inhibitory concentration of nystatin up to 8 times in single biofilm of Candida albicans, and 2 times in mixed species biofilm. A small decrease in the adhesion of Candida tropicalis in NOK Si cells was showed after treatment with terpinen-4-ol, and nystatin had a greater effect for both species. For enzymatic activity, the drugs showed no action. The effect potentiated by the combination of terpinen-4-ol and nystatin and the reduction of adhesion provide evidence of its potential as an anti-fungal agent.

Resumo O objetivo desse estudo foi avaliar a atividade antifúngica do Terpinen4-ol associado à nistatina em biofilmes simples e misto, formados por Candida albicans e Candida tropicalis, bem como o efeito do terpinen-4-ol na adesão em células orais e atividade enzimática. As concentrações inibitórias mínimas e as concentrações fungicidas mínimas do terpinen-4-ol e da nistatina em Candida albicans e Candida tropicalis foram determinadas pelo método de microdiluição em caldo, juntamente com a atividade sinérgica (método do tabuleiro de "xadrez"). Biofilmes simples e misto foram preparados usando o modelo de placa de microtitulação estática e quantificados por unidades formadoras de colônias (CFU/mL). O efeito do Terpinen-4-ol na adesão de Candida albicans e Candida tropicalis em co-cultura com queratinócitos orais (NOK Si) foi avaliado, bem como a atividade enzimática, medindo o tamanho da zona de precipitação, após o crescimento em ágar fosfolipase, protease e hemolisina. O terpinen-4-ol (4.53 mg mL-1) e a nistatina (0,008 mg mL-1) conseguiram inibir o crescimento de biofilmes e um efeito antifúngico sinérgico foi demonstrado com a associação de fármaco, reduzindo a concentração inibidora de nistatina até 8 vezes em biofilme simpes de Candida albicans e 2 vezes em biofilme misto. Uma pequena diminuição na adesão de Candida tropicalis em células NOK Si foi mostrada após o tratamento com terpinen-4-ol e a nistatina teve um efeito maior para ambas as espécies. Para a atividade enzimática, as drogas não apresentaram ação. O efeito potencializado pela combinação de terpinen-4-ol e nistatina e a redução de adesão evidenciam seu potencial como agente anti-fúngico.

Atividade anti-Candida tropicalis dos enantiômeros (R)-(+) ­ & (S)-(-) ­ citronelal em associação com cetoconazol / Anti-Candida tropicalis activity of (R)-(+) ­ & (S)-(-) ­ citronellal enantiomers in combination with ketoconazole

Introdução: aproximadamente 75% das mulheres saudáveis experimentam pelo menos um episódio sintomático de candidíase vulvovaginal (CVV) durante sua vida. Objetivo: avaliar a atividade antifúngica contra cepas de C. tropicalis dos enantiômeros (R)-(+) ­ e (S)-(-)-citronelal [(R)-(+) ­ e (S)-(-)-CT] em associação com cetoconazol. Metodologia: o efeito antifúngico de ambos os enantiômeros foram quantificados e classificados como fungicida ou fungistático a partir dos resultados obtidos da microdiluição em meio líquido RPMI1640 para a obtenção da concentração inibitória mínima (CIM) e da concentração fungicida mínima (CFM). Foram realizados ensaios de associação do antifúngico padrão, cetoconazol com os fitoconstituintes por difusão em Agar e os resultados foram classificados como sinérgicos, antagônicos e indiferentes. Resultados: a CIM50 e a CFM50 dos compostos (R)-(+) ­ e (S)-(-)-citronelal foram respectivamente 16 e 64µg/mL e 2×CIM. Houve sinergismo para todas as cepas testadas com ambos os compostos, porém com maior efeito do enantiômero (S)-(-)-CT sobre as cepas LM 665 e LM 255 em relação ao enantiômero (R)-(+)-CT. Conclusão: os compostos naturais deste estudo mostraram efeito fungicida sobre as cepas testadas, bem como efeito sinérgico significativo quando associado ao cetoconazol

Investigation of drug resistance and multilocus sequence typing for differentiation of strains of Candida tropicalis of Shanghai / 中华预防医学杂志

Objective@#To analyze the drug resistance of clinical isolates of Candida tropicalis in patients with infectious diseases, and preliminarily study their molecular characteristics.@*Methods@#95 strains of Candida tropicalis were isolated from the fungal culture specimens of 87 patients with infectious diseases in Shanghai Public Health Clinical Center from 2012 to 2015. Meanwhile, basic clinical data of patients were collected. The drug resistance of the strains to fungal drugs was analyzed by ATB FUNGUS 3 drug sensitivity test strips. All strains were classified by Multilocus sequence typing(MLST). Then, homology analysis was conducted by MEGA 5.2 software, and the evolutionary tree was mapped by using UPGMA method.@*Results@#Patients distribution of strains was rendered as following: 31 strains from TB patients, 21 strains from HIV/AIDS patients, 19 strains from patients with liver disease, and 24 strains from rare cause infection or fever patients. The drug resistance rate to five antifungal drugs commonly used in clinical (amphotericin B, 5-fluorine cytosine, fluconazole, itraconazole, voriconazole) were 2.11% (2 strains), 0, 26.32% (25 strains), 26.32% (25 strains), and 26.32% (25 strains) respectively. Among the 25 azole-resistant strains: 14 strains were from rare cause infection or fever patients, 8 strains were from HIV/AIDS patients, and 3 strains were from tuberculosis patients. In MLST, 72 sequence types (ST types) were produced, 70 of which were new types. Evolutionary tree analysis showed that 95 strains of clinical strains distribute as three large clusters. 24 azole resistant strains (96.0%) were located in CLUSER Ⅰ.@*Conclusion@#The isolated Candida tropicalis were mainly resistant to azole drugs. MLST typing indicates that they was closely related to their genetic background.

Fungoma vesical por Candida tropicalis: un caso clínico pediátrico / Bladder fungus ball by Candida tropicalis: a pediatric case report

Vesical fungus ball is a mobile, oval and echogenic mass as a result of accumulation of long and wide numerous hyphae. Fungal urinary tract infection incidence has increased notoriously and there are isolated yeast in 7 to 8% of urine cultures. Different species of Candida are cause of urinary tract infection. Epidemiologically, the first isolated pathogen is Candida albicans, followed by Candida tropicalis. Bladder poll has been documented as the most important risk factor for candiduria in critical patients into intensive care.

Un fungoma vesical es una masa móvil, oval y ecogénica en la vejiga resultante del acúmulo de hifas largas y anchas. La incidencia de la infección urinaria de etiología fúngica se ha incrementado notablemente. Se aíslan levaduras en 7 a 8% de los urocultivos. Diferentes especies de Candida son causantes de infección urinaria, siendo Candida albicans la más frecuente, seguida de Candida tropicalis. Presentamos el caso de un niño varón de cuatro años, con un síndrome de Guillain Barré, catéter urinario permanente, estadía prolongada en UCI y expuesto a tratamiento antibacteriano de amplio espectro que desarrolló un fungoma vesical, diagnosticado por ecotomografía, con aislamiento de C. tropicalis en orina. Se trató con anfotericina B deoxicolato y extracción del fungoma por cistoscopia, con buena respuesta clínica. El cateterismo vesical se ha documentado como el factor de riesgo más importante para candiduria en pacientes de terapia intensiva.

Comparative analysis on candida infection detection methods of vaginal secretion / 重庆医学

Objective To analyze the correlation among 3 kinds of detection methods of routine microscopic examination,fluorescence PCR nucleic acid amplification and fungal color culture in the fungal detection of vaginal secretion.Methods The patients with suspected vaginitis treated in this hospital during 2014-2016 were selected.Each 500 cases of negative and positive vaginal secretion samples by microscopic examination were collected.The candida types were identified by using the fluorescence PCR nucleic acid amplification,then 100 samples with the positive results of fluorescence PCR nucleic acid amplification for detecting fungal were performed the fungal microbial culture to verify the accuracy rate of typing results.Results The Kappa value of consistency test between fluorescence PCR nucleic acid amplification and routine microscopic examination was 0.632,the consistency between them was poor.Among 500 positive samples of vaginal secretion detected by routine microscopic examination,382 cases (76.4 ％) of Candida albicans infection were detected by fluorescence PCR nucleic acid amplification,73 cases (14.6％) were Candida glabrata infection,10 cases (2.0 ％) were Candida tropicalis infection,3 cases (0.6 ％) were Candida albicans combined Candida glabrata infection and 32 cases (6.4 ％) were other fungal infection.Among 500 negative samples by conventional microscopic examination,152 positive cases were identified by fluorescence PCR nucleic acid amplification,including 130 cases of Candida albicans,16 cases of Candida glabrata and 6 cases of Candida tropicalis.There was no statistical significant difference in positive rate between the fluorescence PCR nucleic acid amplification and CHROMAgar rapid color method (x2 =0.131,P =0.936).Conclusion For the patients with clinical manifestations and negative microscopic examination results,it is recommended to conduct fluorescence PCR nucleic acid amplification rapid type identification or fungal culture identification.

Effect of ctpxa1 gene deletion in Candida tropicalis on long chain dicarboxylic acid accumulation / 生物工程学报

Candida tropicalis uses alkanes and fatty acids to produce long chain dicarboxylic acids. However, the yield can be affected by β-oxidation in peroxisomes. Pxa1p was a membrane protein of Saccharomyces cerevisiae peroxisomes. Pxa1p and Pxa2p form a dimer that is involved in transporting of long chain fatty acids into peroxisomes, but the similar transporting system of Candida tropicalis has not yet been reported. In this study, a ctpxa1 gene deletion strain named C. tropicalis 1798-pxa1 was constructed by homologous single exchange method using PCR fragment. The expression of ctpxa1 gene in C. tropicalis 1798, C. tropicalis 1798-pxa1 was detected by semi-quantitative RT-PCR, and the ratio of gray value was 2.03, implying that the expression of ctpxa1 in C. tropicalis 1798-pxa1 was weakened. After 144 h fermentation, the dodecanedioic acid production of C. tropicalis 1798-pxa1 was increased 94.3% than the former strain, the maximum yield was 10.3 g/L.

Evaluación de la portación de cándida spp en pinzas ortodóncicas post utilización / Evaluation of candida spp carriage on orthodontic pliers, after using them

Objetivo: evaluar el nivel de contaminación por Candida spp, post uso del instrumental de ortodoncia que se utiliza intrabucalmente en pacientesColombianos y Argentinos. Materiales y métodos: Se incluyeron pacientesentre 16 y 65 años, de ambos sexos, con armado de brackets superior e inferior. Criterios de Exclusión: pacientes con brackets de autoligado, con enfermedades autoinmunes, con enfermedades sistémicas con manifestaciones en el periodonto, pacientes fumadores, menores de 16 años y con armado de brackets en un solo maxilar. Se analizaron los alicates de corte distal Hu Friedy luego de ser utilizados para seccionar el arco por distal del último tubo presente enboca. Se estudiaron 80 pacientes agrupados en: Grupo A 40 pacientesColombianos, Grupo B 40 pacientes argentinos. Se realizó una primer tomaintrabucal con un hisopo estéril y se llevó a un tubo con medio de Stuart. Se cortaron las 4 secciones distales de los arcos en boca tardando al menos 1 minuto. Con un segundo hisopo estéril se frota toda la superficie del alicate post uso y se llevaron a otro tubo. Se sembraron en Chromagar Candida y se realizaron pruebas microbiológicas convencionales. Resultados: Las pinzas estériles aparecen contaminadas post corte distal delos alambres en el 95% de los casos en ambos países. Hubo diferencia entrelos pacientes colombianos y los argentinos respecto a las especies, ya que enlos primeros la especie con mayor prevalencia fue Candida tropicalis tanto en lapinza como en la cavidad bucal, mientras que en los argentinos fue Candidaalbicans. Conclusión: las pinzas de ortodoncias se contaminan con Candidaspp post utilización en boca y son un fómite a partir del cual puede generarsediseminación sistémica así como infección cruzada.

Practice of Clinical Pharmacists Participating in Anti-infective Treatment for A Patient with Tropical Candi-demia / 中国药房

OBJECTIVE:To explore the role of clinical pharmacists participating in drug therapy for patients with severe infec-tions. METHODS:Clinical pharmacists participated in drug treatment for a patient with tropical candidemia and assisted physicians to adjust anti-infective treatment plan. According to the results of blood culture,clinical pharmacists suggested Piperacillin sodium and tazobactam sodium for injection 3.75 g,ivgtt,q8 h+Caspofungin acetate for injection 50 mg (initial dose of 70 mg),ivgtt, qd,for symptomatic treatment;increased the daily dose of Caspofungin acetate for injection to 50 mg,ivgtt,bid due to plasma ex-change;Caspofungin acetate for injection 50 mg,ivgtt,qd+Amphotericin B for injection 0.1 mg/kg,ivgtt,qd for anti-infective plan due to the possible“contradiction”of echinocandins;closely monitored ADR,such as allergy,erythra,renal function injury. RE-SULTS:Physicians adopted the suggestions of clinical pharmacists,vital sign of patient kept stable,and tropical candidemia was not detected in the blood culture;the patient was transferred to general ward for further treatment. CONCLUSIONS:Based on the results of blood calture,clinical symptoms and the characteristics of drug effects,clinical pharmacists participated in the treatment for the patient with severe infection,retrieved related treatment guideline,assisted physicians to adjust anti-infective plan and close-ly monitored possible ADR so as to guarantee the effectiveness and safety of anti-infective treatment.

Mixed biofilms formed by C. albicans and non-albicans species: a study of microbial interactions

Abstract Most Candida infections are related to microbial biofilms often formed by the association of different species. The objective of this study was to evaluate the interactions between Candida albicans and non-albicans species in biofilms formed in vitro. The non-albicans species studied were:Candida tropicalis, Candida glabrata andCandida krusei. Single and mixed biofilms (formed by clinical isolates of C. albicans and non-albicans species) were developed from standardized suspensions of each strain (107 cells/mL), on flat-bottom 96-well microtiter plates for 48 hour. These biofilms were analyzed by counting colony-forming units (CFU/mL) in Candida HiChrome agar and by determining cell viability, using the XTT 2,3-bis (2-methoxy-4-nitro-5-sulphophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide colorimetric assay. The results for both the CFU/mL count and the XTT colorimetric assay showed that all the species studied were capable of forming high levels of in vitro biofilm. The number of CFU/mL and the metabolic activity of C. albicans were reduced in mixed biofilms with non-albicans species, as compared with a singleC. albicans biofilm. Among the species tested, C. krusei exerted the highest inhibitory action against C. albicans. In conclusion, C. albicans established antagonistic interactions with non-albicans Candida species in mixed biofilms.

Identificación de Candida glabrata y otras especies comunes del género Candida mediante el uso secuencial del medio de cultivo cromógeno y la prueba del tubo germinal / Identification of Candida glabrata and other common Candida species using chromogenic agar and germ tube test / Identificação de Candida glabrata e outras espécies comuns do gênero Candida mediante o uso sequencial do meio de cultivo cromogênico e a prova do tubo germinal

Introdución: se han usado agar cromógeno (AC) y la prueba del tubo germinal (PTG) para identificar C. albicans. Sin embargo, ninguna de estas dos pruebas por separado permite la identificación de C. glabrata. Objetivo: analizar la eficacia diagnóstica del uso secuencial del AC y la PTG para identificar las especies más comunes de Candida. Métodos: se identificaron 436 aislamientos usando el AC y luego la PTG; se utilizaron las pruebas bioquímicas como estándar de oro, y se determinaron la sensibilidad y especificidad del esquema secuencial con sus intervalos de confianza. Resultados: el uso en serie del AC y la PTG tuvo sensibilidad del 97,9 % (IC95 %: 96,0-99,9) para identificar C. albicans/dubliniensis y del 90,9 % (IC95 %: 84,0-97,8) para identificar C. tropicalis, con especificidad del 100 % para ambas especies. El mismo esquema permitió identificar C. glabrata con sensibilidad del 92,5 % (IC95 %: 86,2- 98,8) y especificidad del 96,6 % (IC95 %: 95,0-98,6), y el complejo C. parapsilosis con especificidad del 96,3 % (IC95 %: 94,5-98,1). Conclusiones: el esquema propuesto permite la identificación de C. albicans/dubliniensis, C. tropicalis y C. glabrata con sensibilidad y especificidad adecuadas, y podría ser útil en entornos clínicos de bajos recursos.

Introduction: Chromogenic agar and germ tube test have been used for decades to identify C. albicans. However, neither of these tests separately allows identification of C. glabrata. Objective: To test the efficacy of chromogenic agar and germ tube test used serially to identify the most common Candida species found in clinical practice, with emphasis on C. glabrata. Methods: 436 isolates were identified using the chromogenic medium followed by the germ tube test. Biochemical tests were used as gold standard. Sensitivity, specificity and confidence intervals (95 %) of the serial identification system were determined. Results: Sensitivity was 97.9 % (IC95 %: 96.0-99.9) to identify C. albicans/dubliniensis, and 90.9 % (IC95 %: 84.0-97.8) for C. tropicalis; specificity was 100 % for both species. Sensitivity was 92.5 % (IC95 %: 86.2-98.8) for identification of C. glabrata with 96.6 % (IC95 %: 95.0-98.6) specificity. Concerning identification of the C. parapsilosis complex, specificity was 96.3 % (IC95 %: 94.5-98.1). Conclusion: The proposed serial scheme has adequate sensitivity and specificity for identification of C. albicans/dubliniensis, C. tropicalis and C. glabrata. It could be useful in low-resources clinical settings.

Se usaram agar cromogênico (AC) e a prova do tubo germinal (PTG) para identificar C. albicans. No entanto, nenhuma destas duas provas por separado permite a identificação de C. glabrata. Objetivo: analisar a eficácia diagnóstica do uso sequencial do AC e a PTG para identificar as espécies mais comuns de Candida. Métodos: identificaram-se 436 isolamentos usando o AC e depois a PTG; utilizaram-se as provas bioquímicas como padrão de ouro, e se determinaram a sensibilidade e especificidade do esquema sequencial com seus intervalos de confiança. Resultados: o uso em série do AC e a PTG teve sensibilidade de 97,9 % (IC95 %: 96,0-99,9) para identificar C. albicans/dubliniensis e de 90,9 % (IC95 %: 84,0-97,8) para identificar C. tropicalis, com especificidade de 100 % para ambas espécies. O mesmo esquema permitiu identificar C. glabrata com sensibilidade de 92,5 % (IC95 %: 86,2- 98,8) e especificidade de 96,6 % (IC95 %: 95,0-98,6), e o complexo C. parapsilose com especificidade de 96,3 % (IC95 %: 94,5-98,1). Conclusões: o esquema proposto permite a identificação de C. albicans/dubliniensis, C. tropicalis e C. glabrata com sensibilidade e especificidade adequadas, e poderia ser útil em meios clínicos de baixos recursos.