Comparative in vitro analysis of the antifungal activity of different calcium silicate-based endodontic sealers

Aim: This study aimed to perform an in vitro comparative analysis of the antifungal activity of different calcium silicate-based endodontic sealers against three fungal species. Methods: The antifungal properties of three calcium silicate-based sealers were tested: Bio-C Sealer, Cambiar a Sealer Plus BC, and MTA-Fillapex. Two commonly used sealers were used as controls: AH Plus and Endomethasone. An agar diffusion test was performed to analyze the antifungal activity of the sealers against Candida albicans, Candida glabrata, Candida tropicalis, and a mixed microbial culture medium. The results were analyzed using ANOVA (p <0.05). Results: Endomethasone exhibited the highest inhibition against all strains examined, maintaining a consistent level of inhibition throughout 7 days. MTA-Fillapex demonstrated the best performance among the calcium silicate-based sealers for the three fungal species (p < 0.05), maintaining stable values over the 7 days, surpassing that of Endomethasone. Nevertheless, MTA-Fillapex only exhibited antimicrobial effect against the mixed culture for the first 24 hours, and no antimicrobial activity was observed at 48 hours, being surpassed by all tested sealers (p < 0.05). Conclusion: Of all silicate-based sealers tested, only MTA-Fillapex exhibited promising antifungal activity. Nevertheless, care must be taken when extrapolating these results, as MTA-Fillapex exhibited poor antimicrobial activity when tested in mixed microbial cultures

Mortalidad de la sepsis en la unidad de cuidados intensivos / Mortality from sepsis in intensive care unit

INTRODUCCIÓN. La sepsis es un estado de disfunción multisistémica, que se produce por una respuesta desregulada del huésped a la infección. Diversos factores influyen en la gravedad, manifestaciones clínicas y progresión de la sepsis, tales como, heterogeneidad inmunológica y regulación dinámica de las vías de señalización celular. La evolución de los pacientes depende del tratamiento oportuno, las escalas de puntuación clínica permiten saber la mortalidad estimada. OBJETIVO. Evaluar la mortalidad en la unidad de cuidados intensivos; establecer el manejo y la utilidad de aplicar paquetes de medidas o "bundlers" para evitar la progresión a disfunción, fallo multiorgánico y muerte. METODOLOGÍA. Modalidad de investigación tipo revisión sistemática. Se realizó una búsqueda bibliográfica en bases de datos como Google académico, Mendeley, ScienceDirect, Pubmed, revistas como New England Journal Medicine, Critical Care, Journal of the American Medical Association, British Medical Journal. Se obtuvo las guías "Sobreviviendo a la sepsis" actualización 2021, 3 guías internacionales, 10 estudios observacionales, 2 estudios multicéntricos, 5 ensayos aleatorizados, 6 revisiones sistémicas, 5 metaanálisis, 1 reporte de caso clínico, 4 artículos con opiniones de expertos y actualizaciones con el tema mortalidad de la sepsis en UCI con un total de 36 artículos científicos. RESULTADOS. La mortalidad de la sepsis en la unidad de cuidados intensivos, fue menor en el hospital oncológico de Guayaquil, seguido de Australia, Alemania, Quito, Francia, Estados Unidos de Norteamérica y Vietnan, La mortalidad más alta se observa en pacientes con enfermedades del tejido conectivo. DISCUSIÓN. La aplicación de los paquetes de medidas o "bundlers" en la sepsis, se asocia con una mejor supervivencia y menores días de estancia hospitalaria. CONCLUSIÓN. Las escalas SOFA, APACHE II y SAPS II ayudan a predecir la mortalidad de forma eficiente, en la detección y el tratamiento temprano en pacientes con enfermedades agudas y de alto riesgo.

INTRODUCTION. Sepsis is a state of multisystem dysfunction, which is caused by a dysregulated host response to infection. Several factors influence the severity, clinical manifestations and progression of sepsis, such as immunological heterogeneity and dynamic regulation of cell signaling pathways. The evolution of patients depends on timely treatment, clinical scoring scales allow to know the estimated mortality. OBJECTIVE. To evaluate mortality in the intensive care unit; to establish the management and usefulness of applying bundlers to prevent progression to dysfunction, multiorgan failure and death. METHODOLOGY. Systematic review type research modality. A bibliographic search was carried out in databases such as Google Scholar, Mendeley, ScienceDirect, Pubmed, journals such as New England Journal Medicine, Critical Care, Journal of the American Medical Association, British Medical Journal. We obtained the guidelines "Surviving Sepsis" update 2021, 3 international guidelines, 10 observational studies, 2 multicenter studies, 5 randomized trials, 6 systemic reviews, 5 meta-analyses, 1 clinical case report, 4 articles with expert opinions and updates on the subject of sepsis mortality in ICU with a total of 36 scientific articles. RESULTS. The mortality of sepsis in the intensive care unit, was lower in the oncological hospital of Guayaquil, followed by Australia, Germany, Quito, France, United States of America and Vietnam, The highest mortality is observed in patients with connective tissue diseases. DISCUSSION. The application of bundlers in sepsis is associated with better survival and shorter days of hospital stay. CONCLUSIONS. The SOFA, APACHE II and SAPS II scales help to predict mortality efficiently in the early detection and treatment of patients with acute and high-risk disease.

Correlation of extracellular enzymes activity of Candida glabrata clinical isolates with in vivo pathogenicity in Galleria mellonella larvae / 中华预防医学杂志

Objective: To explore the relationship between extracellular enzymes activity and virulence of Candida glabrata clinical isolates based on the infection model of Galleria mellonella larvae. Methods: Using experimental research methods, 71 strains of non-repetitive Candida glabrata were collected from Qinghai Provincial People's Hospital from June 2021 to January 2022. Bovine serum protein agar medium, egg yolk agar medium, sheep blood agar medium, Tween-80 agar medium and triglyceride agar medium were used to detect the aspartyl protease activity, phospholipase activity, hemolysis activity, esterase activity and lipase activity of Candida glabrata. Median lethal concentration (LC50) was calculated by using 1.25×108 CFU/ml,2.50×108 CFU/ml,3.75×108 CFU/ml,5.00×108 CFU/ml suspension of Candida glabrata ATCC2001 to infect Galleria mellonella larvae. Histopathological and etiological analysis was performed to determine whether the infection model was successfully established. The clinical isolates of Candida glabrata were configured to infect Galleria mellonella larvae with LC50 concentration to detect the pathogenicity of Galleria mellonella larvae.Spearman test or Pearson test were used to analyze the correlation between the extracellular enzyme activity of Candida glabrata clinical isolates and the pathogenicity of Galleria mellonella larvae. Results: 71 strains of Candida glabrata isolated clinically were detected to have low hemolytic activity after 2 days of culture. Aspartyl protease was detected after 4 days of culture, among which 7 strains (9.86%), 19 strains (26.76%) and 45 strains (63.38%) showed low, medium and high aspartyl protease activity. After 7 days of culture, 71 strains did not detect phospholipase, esterase and lipase activities. Candida glabrata on Galleria mellonella larvae of LC50=2.5×108 CFU/ml Fungal spore were found in the intestinal tissue pathological section of Galleria mellonella larvae in the experimental group, and Candida glabrata was identified by the microbial Mass Spectrometry after culture, while no fungi were found in the pathological section and culture of the control group. Spearman test shows that, there was a linear positive correlation between aspartyl protease activity and the survival rate of Galleria mellonella larvae (r = 0.73, P<0.01), the difference was statistically significant.Pearson test shows that, there was no significant linear relationship between hemolytic activity and survival rate of Galleria mellonella larvae (r = 0.16, P = 0.34), the difference was not statistically significant. Conclusion: The clinical isolates of Candida glabrata in this study had aspartyl protease activity and low hemolytic activity, but no phospholipase, esterase and lipase activity. The activity of aspartyl aspartyl protease of Candida glabrata was positively correlated with the pathogenicity of Galleria mellonella larvae.

Avaliação in vitro de formulações do óleo essencial de citronela (Cymbopogon nardus) sobre propriedades biológicas, físicas e mecânicas de resina acrílica para prótese dentária / In vitro evaluation of citronella (Cymbopogon nardus) essential oil formulations on biological, physical and mechanical properties of acrylic resin for dental prosthesis

O controle de formação de biofilme deve ser promovido por meio de técnicas diárias de higienização, sendo uma delas o uso de soluções de desinfecção. Dentre as soluções químicas comercialmente disponíveis, algumas podem ocasionar efeitos adversos quando utilizadas em longo prazo. Dessa forma, a busca por métodos alternativos de desinfecção, com soluções que não alterem as propriedades do material e que sejam inertes para o seu usuário torna-se essencial. Este estudo teve como objetivos: avaliar in vitro, a ação antifúngica de formulações de 5 mg/mL à base do óleo de citronela em biofilmes mistos de espécie de Candida em resina acrílica ativada termicamente (RAAT) para base protética por meio de contagem do número de unidades formadoras de colônias (UFC/mL), em dois tempos de exposição (15 e 30 minutos) e avaliar a citotoxicidade das formulações em células da linhagem epitelial HaCat, e verificar a ação destas formulações de citronela a 5 mg/mL na alteração de propriedades físicas e mecânicas de amostras de RAAT. Inicialmente, foram feitas as formulações à base de citronela em forma de emulsão e nanoemulsão. Em seguida, por meio de ensaios de microdiluição em caldo, foi obtida a concentração inibitória mínima (CIM) e concentração fungicida mínima (CFM) das cepas de Candida testadas. Foram confeccionadas amostras de RAAT para a realização dos ensaios, sendo 96 amostras para o ensaio de biofilme, divididas de acordo com os grupos: GI - controle negativo (Biofilme misto C. albicans ATCC + C. glabrata ATCC + C. albicans oral 6892 + C. albicans oral 7037); GII- Nanoemulsão de Citronela 5mg/mL; GIIIControle da Nanoemulsão (Tween 20); GIV- Emulsão de Citronela 5mg/mL; GV- Controle da Emulsão (Goma Xantana); GVI- Clorexidina 0,12% (Periogard), e 20 para o ensaio de viabilidade celular, divididas nos grupos: GI- Nanoemulsão de Citronela 5mg/mL; GII- Controle da Nanoemulsão (Tween 20); GIII- Emulsão de Citronela 5mg/mL; GIV- Controle da Emulsão (Goma Xantana); GV- Clorexidina 0,12% (Periogard). Biofilmes mistos de Candida foram formados sobre as superfícies das amostras, e foram quantificados por meio de contagem de unidades formadoras de colônias (UFCs). Para o teste de viabilidade celular utilizou-se células epiteliais da linhagem HaCaT, avaliadas através da coloração com resazurina. Para avaliação das propriedades físicas e mecânicas foram confeccionadas 50 amostras retangulares e 50 circulares de RAAT, separadas aleatoriamente em 5 grupos, de acordo com a formulação utilizada: GI ­ controle (água destilada); GII ­ saliva artificial; GIII ­ nanoemulsão de citronela a 5mg/mL; GIV - emulsão de citronela a 5mg/mL e GV - Clorexidina 0,12% (Periogard). As amostras foram imersas nas respectivas formulações, simulando a desinfecção de próteses dentarias pelo período de 30 minutos diários, durante 6 meses. Após este período, foi feita a análise da alteração de rugosidade de superfície (Ra - µm), estabilidade de cor (CIEDE 2000), avaliação da microdureza Knoop e resistência a flexão (MPa) das amostras. Os dados obtidos foram submetidos às análises estatísticas (p< 0,05). As formulações fitoterápicas à base de citronela apresentaram valores de CIM e CFM que variaram entre 0,156-1 mg/mL sobre as espécies de Candida e inibiram o crescimento do biofilme misto, apresentando uma redução estatisticamente significativa na contagem de UFC/mL, de até 2,7 Log destes fungos, principalmente quando submetidas ao tempo de exposição de 30 minutos. As formulações fitoterápicas testadas não apresentaram diferenças estatísticas entre si, apresentando viabilidade da célula HaCaT maior que 70%, diferentemente da clorexidina a 0,12%. Não houve diferença nos valores médios de rugosidade de superfície em nenhum dos grupos após serem submetidos à imersão nas formulações testes, com exceção do grupo controle, que apresentou diferença estatística significativa na rugosidade de superfície após o período de imersão. Nos valores de alteração de cor (ΔE00) foi possível observar que não houve diferença estatística significativa entre os grupos experimentais antes e após serem submetidos a imersão nas formulações testes. Nos valores de microdureza Knoop houve diferença estatística significativa apenas entre o grupo controle e o grupo emulsão de citronela. Para os dados de resistência flexural, módulo de elasticidade, força máxima e alongamento não houve diferença estatística significativa entre os grupos após imersão. Em suma, concluiu-se que as formulações fitoterápicas à base de citronela apresentaram efeito antifúngico sobre as diferentes espécies de Candida em superfície de resina acrílica para próteses dentárias, não apresentaram efeito citotóxico sobre células da linhagem epitelial, e se mostraram seguras como formulações antissépticas no que se refere às propriedades físicas e mecânicas estudadas, sendo potencialmente promissoras para serem indicadas como soluções desinfetantes para próteses dentárias(AU)

The control of biofilm formation should be promoted through daily hygiene techniques such as the use of disinfection solutions. Among the commercially available chemical solutions, some can lead to adverse effects in long-term use. Thus, the search for alternative methods of disinfection with solutions that do not interfere with properties of the material and those inert to the user becomes essential. This study aimed to: evaluate, in vitro, the antifungal effect of 5 mg/mL formulations based on citronella oil on mixed biofilms of Candida species in thermally activated acrylic resin (TAAR) for prosthetic base by counting the number of colony forming units (CFU/mL), in two exposure times (15 and 30 minutes) and to evaluate the cytotoxicity of the formulations in cells of the HaCat epithelial lineage, and to verify the action of these formulations of citronella at 5 mg/mL in the alteration of physical and mechanical properties of TAAR samples. Initially, citronella-based formulations were prepared as emulsion and nanoemulsion. Then, by means of broth microdilution assays, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the tested Candida strains were obtained. For the biofilm test, 96 TAAR samples were prepared and assigned to the groups, as follows: GI - negative control (Mixed biofilm C. albicans ATCC + C. glabrata ATCC + C. albicans oral 6892 + C. albicans oral 7037); GII- Citronella Nanoemulsion 5mg/mL; GIII- Nanoemulsion Control (Tween 20); GIV- Citronella Emulsion 5mg/mL; GVEmulsion Control (Xanthan Gum); GVI- Chlorhexidine 0,12% (Periogard). For the cell viability assay, 20 samples were prepared and assigned to the following groups: GI- Citronella Nanoemulsion 5mg/mL; GII- Nanoemulsion Control (Tween 20); GIII- Citronella Emulsion 5mg/mL; GIV- Emulsion Control (Xanthan Gum); GV- Chlorhexidine 0,12% (Periogard). Mixed Candida biofilms were formed on the surfaces of the samples, and were quantified by counting colony forming units (CFUs). For the cell viability test, epithelial cells of the HaCaT lineage were used, evaluated by staining with resazurin. To evaluate the physical and mechanical properties, 50 rectangular and 50 circular samples of TAAR were prepared and randomly assigned into 5 groups, according to the formulation used: GI ­ control (distilled water); GII ­ artificial saliva; GIII ­ citronella nanoemulsion at 5mg/mL; GIV ­ citronella emulsion at 5mg/mL and GV ­ chlorhexidine 0,12% (Periogard). The samples were immersed in the respective formulations, simulating the disinfection of dental prostheses for a period of 30 minutes daily, for 6 months. After this period, the analysis of surface roughness change (Ra - µm), color stability (CIEDE 2000), evaluation of Knoop microhardness and flexural strength (MPa) of the samples was performed. The data obtained were submitted to statistical analysis (p< 0,05). The phytotherapic formulations based on citronella presented MIC and MFC values that varied between 0,156-1 mg/mL on Candida species and inhibited the growth of mixed biofilm, showing a statistically significant reduction in the CFU/mL counts, up to 2,7 Log of these fungi, mainly when subjected to an exposure time of 30 minutes. The phytotherapic formulations tested did not show statistical differences between them, showing HaCaT cell viability greater than 70%, unlike 0,12% chlorhexidine. There was no difference in the mean values of surface roughness in any of the groups after being subjected to immersion in the test formulations, except for the control group, that presented a statistically significant difference in surface roughness after the immersion period. Regarding color change (ΔE00) it was possible to observe that there was no statistically significant difference between the experimental groups before and after being subjected to immersion in the test formulations. In the Knoop microhardness values, there was a statistically significant difference only between the control group and the citronella emulsion group. For data on flexural strength, modulus of elasticity, maximum force and stretching, there was no statistically significant difference between the groups after immersion. In summary, it can be concluded that phytotherapic formulations based on citronella had an antifungal effect on different Candida species on acrylic resin surfaces for dental prostheses, besides not presenting a cytotoxic effect on cells of the epithelial lineage, and proved to be safe as antiseptic formulations in the which refers to the physical and mechanical properties studied, being potentially promising to be indicated as disinfectant solutions for dental prostheses(AU)

Synergistic interaction of fluconazole/sodium bicarbonate on the inhibition of Candida glabrata phospholipase gene

Abstract Candida glabrata infections are responsible for deaths of people globally. Fluconazole is known to be less effective against C. glabrata, which developed many strategies to evade being destroyed by fluconazole. To achieve enhanced efficacy of fluconazole against C. glabrata, the interaction of fluconazole with sodium bicarbonate was investigated using the CLSI guidelines. The efficacy of fluconazole alone and in combination with sodium bicarbonate was evaluated using the time-kill and phospholipase production assays. Eventually, the expression of PLB was assessed using semi-quantitative RT-PCR to investigate the inhibitory properties of fluconazole alone and in combination with sodium bicarbonate against C. glabrata. The fluconazole/sodium bicarbonate combination displayed synergistic and antagonistic effects (FICI= 0.375-4.25). In C. glabrata ATCC, SN 152, and SN 164, the fluconazole/sodium bicarbonate combination exhibited a significant fungicidal activity (p< 0.05) but antagonistic effect in the case of SN 283. With exception of SN 283, a significant reduction was noted in phospholipase production in clinical isolates of C. glabrata treated with fluconazole/sodium bicarbonate combination. The PLB was down-regulated significantly by 0.168-0.515 fold in C. glabrata treated with fluconazole/sodium bicarbonate. The results suggested fluconazole/sodium bicarbonate to have a potential synergistic interaction in C. glabrata, and the underlying mechanism may be associated with phospholipase gene

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)

Avaliação in vitro da formação de biofilme por cepas de Candida spp. provenientes de indivíduos diabéticos e não diabéticos em ambiente sem e com suplementação de glicose / In vitro evaluation of biofilm formation by Candida spp. strains from diabetic and non-diabetic individuals in environment without and with glucose supplementation

É notório o papel do diabetes mellitus como fator de risco para o desenvolvimento de doenças bucais, como a candidíase bucal, cárie e a periodontite. Assim, este estudo tem como objetivo avaliar a formação de biofilme por cepas de Candida spp. provenientes de diabéticos e não diabéticos em ambiente sem e com suplementação de glicose. Trata-se de um estudo experimental laboratorial in vitro, em três etapas. Etapas um e dois, de obtenção e identificação de 48 cepas de Candida spp., sendo que 32 de C. albicans e 16 de C. glabrata, com auxílio da técnica de PCR. Ainda, a etapa três, de processamento microbiológico, com a avaliação da capacidade de formação de biofilme por três ensaios distintos: I) determinação do número de unidades formadoras de colônia (UFC/mL); II) matéria seca dos biofilmes; III) taxa de crescimento de biofilme em fundo de placa de poliestireno. Inicialmente, objetivando simular as características observadas in vivo, o fundo das placas de cultivo recebeu 400 µL de saliva humana para formação da película adquirida. Decorrida a incubação a 37 °C por 24 h, a saliva foi descartada e cada poço de cultura recebeu suspensão padronizada das leveduras (106 UFC/mL) em Saubouraud Dextrose Broth sem suplementação e com suplementação de glicose a 2 e 10 mg/mL, e as placas foram incubadas a 37 °C por 48 h. Para avaliação do número de UFC/mL, o biofilme aderido foi coletado, diluído seriamente e cultivado em placas de Petri com Sabouraud Dextrose Agar. Após incubação os resultados foram expressos em log UFC/mL. Para a avaliação da matéria seca, a solução remanescente foi liofilizada e mensurada em balança de precisão. A taxa de crescimento de biofilme foi avaliada por microscopia Operetta CLS High Content e o FilmTracer(TM) LIVE/DEAD Biofilm Viability kit, conforme o protocolo do fabricante. Posteriormente, 10 imagens por poço foram obtidas e digitalizadas com ampliação de 40 ×. A área recoberta por biofilme (µm2) das imagens foi avaliada com auxílio do software Harmony High Content Imaging. Os dados apresentaram distribuição não normal, e a comparação entre as cepas de diabéticos e não diabéticos foi realizada pelo teste U Mann-Whitney. O teste de Kruskal-Wallis one way foi utilizado para verificar diferenças entre as condições de suplementação de glicose. O nível de significância estatística adotado foi de α = 5%. Os valores de UFC/mL mostraram um maior crescimento das cepas de C. albicans dos pacientes diabéticos em relação aos não diabéticos nas três suplementações (p < 0,001). Por outro lado, acerca da matéria seca em 10 mg/mL e da taxa de crescimento de biofilme sem suplementação de glicose e a 2 mg/mL, os resultados indicaram uma formação de biofilme maior para cepas de C. albicans dos não diabéticos (p < 0,001). Em conclusão, cepas de C. albicans e C. glabrata provenientes de diabéticos e não diabéticos em ambiente sem e com suplementação de glicose apresentaram resultados distintos quanto à formação de biofilme, por diferentes técnicas

The role of diabetes mellitus is notorious as a risk factor for development of oral diseases, such as oral candidiasis, dental caries, and periodontitis. Thus, this study aimed to evaluate biofilm formation by Candida spp. strains from diabetic and non-diabetic individuals in environment without and with glucose supplementation. This is an in vitro experimental laboratory study, in three stages. Stages one and two of obtainment and identification of 48 Candida spp. strains, with 32 of C. albicans and 16 of C. glabrata, with the help of PCR technique. Also, stage three, of microbiological processing, with evaluation of biofilm formation capacity by three different assays: I) determination of the number of colony forming units (CFU/mL); II) biofilm dry matter; III) biofilm growth rate on the bottom of polystyrene plates. Initially, aiming to simulate the characteristics observed in vivo, the bottom of the cultivation plates received 400 µL of human saliva for formation of acquired pellicle. After the incubation at 37 °C for 24 h, the saliva was discarded and each culture well received standardized suspension of yeast (106 CFU/mL) in Saubouraud Dextrose Broth without supplementation and with glucose supplementation at 2 and 10 mg/mL, and the plates were incubated at 37 °C for 48 h. To assess the number of CFU/mL, the adhered biofilm was collected, seriously diluted, and cultivated in Petri dishes with Sabouraud Dextrose Agar. After the incubation, the results were expressed in log CFU/mL. To assess the dry matter, the remaining solution was lyophilized and measured on a precision scale. The biofilm growth rate was evaluated by Operetta CLS High Content microscopy and FilmTracer(TM) LIVE/DEAD Biofilm Viability kit, according to manufacturer's protocol. Later, 10 images per well were obtained and digitalized with 40 × magnification. The area covered by biofilm (µm2) of the images was assessed with the help of Harmony High Content Imaging software. Data showed non-normal distribution, and the comparison among the diabetic and non-diabetic strains was performed by Mann-Whitney U test. Kruskal-Wallis one-way test was used to verify differences between conditions of glucose supplementation. The level of statistical significance adopted was α = 5%. The values of CFU/mL showed greater growth of the diabetic patient's strains in relation to the non-diabetic ones (p < 0.001). On the other hand, regarding dry matter at 10 mg/mL and the growth rate of biofilm without glucose supplementation and at 2 mg/mL, the results indicated a higher biofilm formation for strains of C. albicans from non-diabetic individuals (p <0.001). In conclusion, C. albicans and C. glabrata strains from diabetic and non-diabetic individuals in environment without and with glucose supplementation showed different results concerning the biofilm formation, using different techniques

Evaluación de susceptibilidad en cándidas spp por colorimetría obtenida en gestantes de un hospital obstétrico / Susceptibility evaluation in candida spp by colorimetry obtained in pregnant women from an obstetric hospital / Avaliação da suscetibilidade em cândida spp por colorimetria obtida em gestantes de um hospital obstétrico

INTRODUCCIÓN: las infecciones fúngicas ocasionadas por levaduras del género Cándida son extremadamente comunes en mujeres de edad reproductiva, y constituyen un motivo de atención medica de salud. OBJETIVO: evaluar la susceptibilidad de Cándidas spp, mediante el método colorimétrico (Integral Yeast System Plus). MÉTODO: fue de tipo descriptivo, transversal; se recopiló información mediante observación directa en campo y el análisis documental para obtener información bibliográfica de tipo secundaria. RESULTADOS: de los 72 casos encontrados de Cándida Albicans revela que son susceptibles a la anfotericina B (2ug/ml); de los 5 casos encontrados de Cándida Krusei revela que son sensibles a la Anfotericina B (2ug/ml); De 1 caso encontrado de Cándida Parapsilosis revela sensibilidad en la Nistatina (1.25ug/ml). En este estudio la prevalencia de la infección por Cándida fue del (44.98%). CONCLUSIONES: Cándida Albicans fue la especie más común aislada en las mujeres embarazadas representando un 72%, En la evaluación de la susceptibilidad a través del kit Integral System Yeast Plus se obtuvo que Cándida Albicans es susceptible a Anfotericina B, Flucitosina entre otros, en Cándida Glabrata se obtuvo que es sensible a la Nistatina, Anfotericina B, susceptible entre otros, en Cándida Krusei se obtuvo que es sensible a la Anfotericina B, Clotrimazol, Miconazol, susceptibles a la Nistatina, Voriconazol y resistente a la Flucitosina, Ketoconazol, Itraconazol y Fluconazol.

INTRODUCTION: fungal infections caused by yeast of the genus Candida are extremely common in women of reproductive age, and constitute a reason for medical health care. OBJECTIVE: to evaluate the susceptibility of Candida spp, using the colorimetric method (Integral Yeast System Plus). METHOD: it was descriptive, transversal; Information was collected through direct observation in the field and documentary analysis to obtain secondary bibliographic information. RESULTS: of the 72 cases found, Candida Albicans reveals that they are susceptible to amphotericin B (2ug / ml); of the 5 cases found, Candida Krusei reveals that they are sensitive to Amphotericin B (2ug / ml); Of 1 case found of Candida Parapsilosis reveals sensitivity in Nystatin (1.25ug / ml). In this study, the prevalence of Candida infection was (44.98%). CONCLUSIONS: Candida Albicans was the most common species isolated in pregnant women, representing 72%. In the evaluation of susceptibility through the Integral System Yeast Plus kit it was obtained that Candida Albicans is susceptible to Amphotericin B, Flucytosine among others, in Candida Glabrata was obtained that it is sensitive to Nystatin, Amphotericin B, susceptible among others, in Candida Krusei it was obtained that it is sensitive to Amphotericin B, Clotrimazole, Miconazole, susceptible to Nystatin, Voriconazole and resistant to Flucytosin, Ketoconazole, Itraconazole and Fluconazole.

INTRODUÇÃO: as infecções fúngicas causadas por leveduras do gênero Candida são extremamente comuns em mulheres em idade reprodutiva e constituem motivo de cuidados médicos. OBJETIVO: avaliar a suscetibilidade de Candida spp, por meio do método colorimétrico (Integral Yeast System Plus). MÉTODO: foi descritivo, transversal; as informações foram coletadas por meio de observação direta em campo e análise documental para obtenção de informações bibliográficas secundárias. RESULTADOS: Dos 72 casos encontrados, Cândida Albicans revelou ser suscetíveis à anfotericina B (2ug /ml); dos 5 casos encontrados, Candida Krusei revela que são sensíveis à Anfotericina B (2ug / ml); de 1 caso encontrado de Candida Parapsilosis revela sensibilidade na Nistatina (1,25ug / ml). Neste estudo, a prevalência de infecção por Candida foi (44,98%). CONCLUSÕES: Cândida Albicans foi a espécie mais comum isolada em gestantes, representando 72%. Na avaliação da susceptibilidade através do kit Integral System Yeast Plus foi obtido que Candida Albicans é suscetível à Anfotericina B, Flucitosina entre outras, em Cândida Glabrata foi obtido que é sensível a Nistatina, Anfotericina B, suscetível entre outras, em Candida Krusei foi obtido que é sensível a Anfotericina B, Clotrimazol, Miconazol, suscetível a Nistatina, Voriconazol e resistente a Flucitosina, Cetoconazol, Itraconazol e Fluconazol.

The secreted acid trehalase encoded by the CgATH1 gene is involved in Candida glabrata virulence

BACKGROUND Candida glabrata yeast is the second cause of candidiasis worldwide. Differs from other yeasts since assimilates only glucose and trehalose (a characteristic used in rapid identification tests for this pathogen) by secreting into the medium a highly active acid trehalase encoded by the CgATH1 gene. OBJECTIVE This study aimed to characterise the function of the acid trehalase in the physiopathology of C. glabrata. METHODS Gene deletion was performed to obtain a mutant ath1Δ strain, and the ability of the ath1Δ strain to grow in trehalase, or the presence of trehalase activity in the ath1Δ yeast cells, was verified. We also tested the virulence of the ath1Δ strain in a murine model of infection. FINDINGS The ath1Δ mutant strain grows normally in the presence of glucose, but loses its ability to grow in trehalose. Due to the high acid trehalase activity present in wild-type cells, the cytoplasmic neutral trehalase activity is only detected in the ath1Δ strain. We also observed a significantly lower virulence of the ath1Δ strain in a murine model of infection with either normal or immunocompromised mice. MAIN CONCLUSIONS The acid trehalase is involved in the hydrolysis of external trehalose by C. glabrata, and the enzyme also plays a major virulence role during infectivity.

Mecanismos de resistencia a fluconazol expresados por Candida glabrata: una situación para considerar en la terapéutica / Resistance mechanisms to fluconazole expressed by Candida glabrata: a situation to consider in therapy / Mecanismos de resistência a fluconazol expressos por Candida glabrata: uma situação a considerar na terapêutica

Introducción: Los esfuerzos terapéuticos orientados a atender las micosis por Candidaspp. se han enfocado en el empleo de azoles; sin embargo, en la literatura científica se discute su beneficio, por los amplios y descritos mecanismos de resistencia. Objetivo: Describir los mecanismos de resistencia al fluconazol expresados por la especie Candida glabrata, con la intención de que sean considerados dentro de las variables de elegibilidad para la intervención. Método: Se realizó una revisión integrativa utilizando la pregunta orientadora: ¿cuáles son los mecanismos de resistencia al fluconazol expresados por la especie Candida glabrata? Veintinueve estudios obtenidos de la base de datos PubMed cumplieron los criterios del análisis crítico propuesto por el instrumento PRISMA, utilizado para la selección de los artículos incluidos para su revisión en este manuscrito. Las categorías bajo las cuales se organizaron los elementos de análisis fueron: sobrexpresión de bombas de eflujo y modificaciones en la enzima lanosterol 14-alfa-desmetilasa. Resultados: Los mecanismos de resistencia al fluconazol expresados por Candida glabrata están determinados principalmente por la regulación a la alza de bombas de adenosina-trifosfato Binding Cassette (ABC) y por la modificación del punto de unión con su blanco farmacológico: la enzima lanosterol 14-alfa-desmetilasa. Conclusión: Los mecanismos de resistencia expresados por Candida glabrata se asocian con la modificación estructural de la diana farmacológica y la sobreexpresión de bombas de eflujo de manera diferencial a otras especies. Se sugiere que Candida glabrata es intrínsecamente menos susceptible al fluconazol.

Introduction: Therapeutic efforts aimed at treating mycosis caused by Candida spp. have focused on the use of azoles; however, their benefits have been subject to discussion in scientific literature, due to the extensive and well-described resistance mechanisms. Objective: To describe the resistance mechanisms to fluconazole expressed by the Candida glabrata species, so they are considered within the variables of eligibility for intervention. Method: An integrative review was carried out using the guiding question: what are the fluconazole resistance mechanisms expressed by the Candida glabrata species? Twenty-nine studies obtained from the PubMed database met the criteria for the critical analysis proposed by the PRISMA instrument, which was used for the selection of articles for review included in this paper. The analysis elements were organized in the following categories: overexpression of efflux pumps and modifications in the enzyme lanosterol 14-alpha-demethylase. Results: The resistance mechanisms to fluconazole expressed by Candida glabrata are mainly determined by the upregulation of Adenosine triphosphate Binding Cassette (ABC) pumps and by the modification of the point of attachment with its pharmacological target: the enzyme lanosterol 14-alpha-demethylase. Conclusion: The resistance mechanisms expressed by Candida glabrata are associated with the structural modification of the pharmacological target and the overexpression of efflux pumps, in a way different to other species. It is suggested that Candida glabrata is intrinsically less susceptible to fluconazole.

Introdução: Os esforços terapêuticos voltados ao tratamento de micose por Candida spp. se focaram no uso de azóis; no entanto, na literatura científica discute-se seu benefício devido aos extensos e descritos mecanismos de resistência. Objetivo: Descrever os mecanismos de resistência ao fluconazol expressos pela espécie Candida glabrata, com a intenção de serem considerados dentro das variáveis de elegibilidade para a intervenção. Método: Uma revisão integrativa foi realizada utilizando a questão norteadora: quais os mecanismos de resistência ao fluconazol expressos pela espécie Candida glabrata? Vinte e nove estudos obtidos da base de dados PubMed atenderam os critérios de análise crítica proposta pelo instrumento PRISMA, utilizado para a seleção dos artigos incluídos para revisão neste manuscrito. As categorias sob as quais se organizaram os elementos de análise foram: superexpressão de bombas de efluxo e modificações na enzima lanosterol 14-alfa-desmetilase. Resultados: Os mecanismos de resistência ao fluconazol expressos por Candida glabrata são determinados principalmente pela regulação positiva das bombas de adenosina-trifosfato Binding Cassette (ABC) e pela modificação do ponto de fixação com seu alvo farmacológico: a enzima lanosterol 14-alfa-desmetilasa. Conclusão: Os mecanismos de resistência expressos por Candida glabrata são associados à modificação estrutural da Diana farmacológica e a superexpressão de bombas de efluxo de maneira diferencial a outras espécies. Sugere-se que Candida glabrata é intrinsecamente menos susceptível ao fluconazol.

Detection of intracellular Helicobacter pylori in Candida. SPP from neonate oral swabs

SUMMARY BACKGROUND: There is evidence of detection of Helicobacter pylori (H. pylori) in the stool of newborns and in the yeast that colonizes the oral cavity of this age group. However, there is a lack of research to confirm it. This study proposes to determine the existence of the bacteria at an early age, specifically in newborns. OBJECTIVE: To identify intracellular H. pylori in oral yeasts and to detect antigens of the bacteria in newborn stools. METHODOLOGY: Cross-sectional and descriptive study. Samples were obtained from infants (oral swab and meconium). Identification of yeast species was performed using the following techniques: CHROMagar Candida, Germinal Tube Test and API Candida Identification System, then the yeasts were observed by light microscopy and fluorescence. Detection of H. pylori antigen in meconium and PCR were performed to amplify specific genes of the bacterium (rRNA16S, cagA, vacA s1a, vacA s1b, vacA s2, vacA m1, vacA m2 and dupA). RESULTS: Intracellular H. pylori was detected in yeast of the species Candida glabrata (C. glabrata) isolated from an oral swab of a newborn. CONCLUSION: The results of this study evidenced the existence of intracellular H. pylori in newborns.

RESUMO ANTECEDENTES: Há evidências de detecçâo de Helicobacter pylori (H. pylori) em fezes de recém-nascidos, como também dentro de leveduras que colonizam a cavidade oral dessa faixa etária. No entanto, faltam investigações que confirmem esses achados. OBJETIVO: Identificar H. pylori intracelular em leveduras de origem oral e detectar antígenos dessa bactéria em fezes neonatais. METODOLOGIA: Estudo transversal e descritivo. As amostras foram obtidas de bebês (zaragatoa oral e mecônio). As identificações das espécies de leveduras foram realizadas utilizando as seguintes técnicas: CHROMagar Candida, teste de tubo germinativo e sistema de identificação API Cândida. As leveduras foram observadas por microscopía óptica e fluorescência. Realizou-se a detecçâo de antígeno de H. pylori em mecônio e PCR para a amplificação de genes específicos desta bactéria (rRNA16S, cagA, vacA s1a, vacA s1b, vacA s2, vacA m1, vacA m2 e dupA). RESULTADOS: Foi detectado H. pylori intracelular em leveduras da espécie Candida glabrata (C. glabrata) isoladas a partir de zaragatoas oral de um recém-nascido. CONCLUSÃO: Os resultados deste estudo evidenciaram a existência interna de levedura de H. pylori em recém-nascidos.

Candidiasis diseminada en paciente inmunocompetente. Revisión bibliográfica a propósito de un caso / Disseminated candidiasis in an immunocompetent patient. Case report and literature review

La candidiasis es la micosis más común entre los pacientes inmunocompetentes de los países desarrollados. Comprende la candidemia, mas frecuente y la candidiasis de los tejidos profundos. La candidiasis de los tejidos profundos tiene una mortalidad de hasta el 40%, incluso en los pacientes que reciben tratamiento oportuno. Además, el desplazamiento global a favor de las especies de cándida no albicans es preocupante, al igual que los nuevos perfiles de resistencia a los antimicóticos actuales.

Candidiasis is the most common mycosis among immunocompetent patients in developed countries. It includes candidemia, more frequent, and candidiasis of deep tissues. Candidiasis of deep tissues has a mortality of up to 40%, even in patients receiving timely treatment. In additioin, the global shift in favor of non-albicans candida species is worrisome, as are the new profiles of resistance to current antifungals.

Estudio clínico y micológico de otomicosis en diabéticos: una serie de 17 casos / Clinical and mycological study of otomycosis in diabetics: series of 17 cases

RESUMEN Introducción: La otomicosis en pacientes inmunosuprimidos esta caracterizada por ser bilateral y ser causada por candida. Pocos estudios comparan las características micológicas encontradas en la microscopía directa y el cultivo. Objetivo: Identificar las características clínicas y micológicas de la otomicosis en pacientes diabéticos. Material y método: Estudio transversal en centro hospitalario de segundo nivel. Criterios de inclusión: pacientes diabéticos con diagnóstico clínico de otomicosis. Intervención: la muestra se examinó directamente bajo el microscopio y se cultivó. Resultados: Se incluyeron 17 pacientes, 10 mujeres y 7 hombres con una edad media de 47,5 años. Los síntomas predominantes fueron hipoacusia en 91,4% (n =16), prurito en 82,4% (n =14), otorrea en 76,5% (n=13)y otalgia en 70,6% (n =12). Afección bilateral se encontró en 47,1% (n =8). Estudio directo al microscopio mostró levaduras en 94,1% (n =16) y 5,9% mostró aspergillus (n =1). Cándida fue el género más comúnmente encontrado en los cultivos y en el examen directo microscópico con 94,1% (n =16) y Candida albicans la especie más común con 88,2% (n =15). Conclusión: Candida albicans es el agente etiológico más común en pacientes diabéticos con otomicosis. Su presentación clínica más frecuente es hipoacusia, prurito y otorrea. El examen directo identificó adecuadamente a los géneros fúngicos.

ABSTRACT Introduction: Otomycosis in immunocompromised patients is characterize by its bilateral course and the predominant etiologic agent is Candida. Few studies compare the mycological features between microscopic direct exam and culture. Aim: To identify the clinical and mycological characteristics of otomycosis in diabetic patients. Material and method: Transversal study. Secondary care center. Inclusion criteria: diabetic patients with clinical diagnosis of otomycosis. Intervention: Direct examination under a microscope of the ear sample and culture. Results: We included 17 patients, 10 women, 7 men with a mean age of 47.5 years. Symptoms were hearing loss 94.1% (n = 16), pruritus 82.4% (n =14) otorrhoea 76.5% (n =13) and otalgia 70.6% (n =12). Bilateral involvement was found in 47.1% (n =8). Direct microscopic study found 94.1% of yeast (n =16) and 5.9% of Aspergillus (n =1). Candida was the most common fungal genus in culture and microscopic exam with 94.1% (n =16) of cases and Candida albicans was the most common species in 88.2% (n =15) cases. Conclusion: Candida albicans is the most common etiologic agent in diabetic patients with otomycosis. Main symptoms were hearing loss, itching and otorrhea. Direct exam correctly identified the fungal genus.

Usefulness of Two-Step Algorithm with Earlier Growth Detection in Anaerobic Bottle and Time to Positivity to Predict Candida glabrata Fungemia / 대한임상미생물학회지

BACKGROUND: Fast identification of Candida glabrata is important, because empirical antifungal therapy for fungemia with C. glabrata and non-C. glabrata varies. We proposed an algorithm for rapid presumptive diagnosis to identify fungemia with C. glabrata using earlier or only growth from anaerobic bottles and longer time to positivity (TTP) in blood cultures. METHODS: Positivity and TTP using the BacT/Alert 3D system (bioMerieux Inc, USA) with resin bottles (FA Plus and FN Plus) were analyzed in 215 candidemia patients from June 2014 to June 2016 in a university-affiliated hospital in Korea. RESULTS: A higher proportion of earlier or only growth from anaerobic bottles was observed in C. glabrata (38.8%, 7/18) than in C. albicans (7.6%, 8/105), C. parapsilosis (10.5%, 4/138), and C. tropicalis (9.2%, 5/54) (P=0.006). The mean (±standard deviation) TTP for C. glabrata was 41.7 h (±16.3 h) compared with 26.7 h (±15.9 h) for C. albicans, 33.4 h (±8.4 h) for C. parapsilosis, and 23.1 h (±17.3 h) for C. tropicalis (P 31.4 h. CONCLUSION: This two-step algorithm in the BacT/Alert 3D system could be the basis for an initial empirical antifungal therapy for fungemia with C. glabrata prior to final identification.

Characterisation of an ABC transporter of a resistant Candida glabrata clinical isolate

BACKGROUND Candida glabrata ranks second in epidemiological surveillance studies, and is considered one of the main human yeast pathogens. Treatment of Candida infections represents a contemporary public health problem due to the limited availability of an antifungal arsenal, toxicity effects and increasing cases of resistance. C. glabrata presents intrinsic fluconazole resistance and is a significant concern in clinical practice and in hospital environments. OBJECTIVE The aim of this study was to characterise the azole resistance mechanism presented by a C. glabrata clinical isolate from a Brazilian university hospital. METHODS Azole susceptibility assays, chemosensitisation, flow cytometry and mass spectrometry were performed. FINDINGS Our study demonstrated extremely high resistance to all azoles tested: fluconazole, voriconazole, posaconazole and itraconazole. This isolate was chemosensitised by FK506, a classical inhibitor of ABC transporters related to azole resistance, and Rhodamine 6G extrusion was observed. A mass spectrometry assay confirmed the ABC protein identification suggesting the probable role of efflux pumps in this resistance phenotype. MAIN CONCLUSIONS This study emphasizes the importance of ABC proteins and their relation to the resistance mechanism in hospital environments and they may be an important target for the development of compounds able to unsettle drug extrusion.

Capacidad de formación de biopelículas en especies del género Candida de procedencia clínica / Capability of biofilm formation in Candida genus species of clinical origin

El objetivo de la investigación fue evaluar la capacidad de formación de biopelículas en especies del género Candida provenientes de muestras clínicas con procesos infecciosos. La muestra estuvo constituida por 70 aislados obtenidos de laboratorios clínicos privados. Para la identificación de las especies se utilizó la Prueba de tubo germinativo y la técnica de microcultivo en agar harina de maíz con tween 80, mientras que la cuantificación de la capacidad de formación de biopelículas se realizó por la técnica de microplaca de poliestireno. Entre los resultados obtenidos en la investigación las especies aisladas fueron: Candida glabrata (34%), Complejo Candida albicans (30%), Complejo Candida parapsilosis (27%) y Candida tropicalis (9%), de las cuales 41 aislados (59%) formaron biopelículas, resultando el Complejo Candida albicans y Candida glabrata las especies con mayor grado de formación de biopelículas con 29% cada una, seguida de Complejo Candida parapsilosis con 27% y Candida tropicalis con 15%. Las cepas que tuvieron mayor formación de biopelículas provenían de orina, seguidas de secreción ótica, secreción vaginal y sangre, sin embargo al aplicar la prueba de Chi-cuadrado (x2) de Pearson se determinó que no existe asociación estadísticamente significativa entre la capacidad de formación de biopelículas y el grado de formación con la procedencia clínica así como tampoco hay asociación entre la capacidad de formación de biopelículas y grado de formación con la especie involucrada (p > 0,05)

The aim of this research was to evaluate the capability of biofilm formation in Candida genus species from clinical samples with infectious conditions. The sample consisted of 70 isolates obtained from private clinical laboratories. Germ tube test and agar cornmeal with tween 80 microculture technique were used as species identification, meanwhile, quantification of biofilm formation capacity was performed by the polystyrene microplate technique. Among the results obtained in the research, isolated species were: Candida glabrata (34%), Candida albicans Complex (30%), Candidaparapsilosis Complex (27%) and Candida tropicalis (9%), of which 41 isolates (59%) formed biofilms, resulting Candida albicans and Candida glabrata Complex species with the highest degree of biofilms with 29% each, followed by Candida parapsilosis Complex with 27% and Candida tropicalis with 15%. Strains with more biofilm formation came from urine, followed by ear discharge, vaginal secretion and blood. However, by applying the Chi squaretest the Pearson, it was determined that there was no statistically significant association between thecapability of biofilm formation and the formation degree with clinical origin, neither there is association between the ability of biofilm formation and the formation degree with the species involved (p> 0.05).

Azole Resistance Caused by Increased Drug Efflux in Candida glabrata Isolated from the Urinary Tract of a Dog with Diabetes Mellitus

A yeast-like organism was isolated from a urine sample of a 6-year-old neutered male miniature poodle dog with urinary tract infection, diabetes ketoacidosis, and acute pancreatitis. We identified the yeast-like organism to be Candida glabrata and found that this fungus was highly resistant to azole antifungal drugs. To understand the mechanism of azole resistance in this isolate, the sequences and expression levels of the genes involved in drug resistance were analyzed. The results of our analysis showed that increased drug efflux, mediated by overexpression of ATP transporter genes CDR1 and PDH1, is the main cause of azole resistance of the C. glabrata isolated here.

Trend of Prevalence and Antifungal Drug Resistance of Candida Species Isolated from Candidemia Patients at a Tertiary Care Hospital During Recent Two Decades / 대한임상미생물학회지

BACKGROUND: Candidemia has increased with an increasing number of people in the high risk group and so has become more important. This study was conducted to investigate the isolation rate of Candida species from candidemia patients and the change in rate of antifungal resistance. METHODS: At a single tertiary care hospital, 1,120 blood cultures positive for Candida species from 1997 to 2016 were investigated according to date of culture, gender, age, and hospital department. RESULTS: During the investigation period, the number of candidemia patients increased from 14 in 1997 to 84 in 2016. The most common organism identified during the two decades was Candida albicans (40.8%), followed by Candida parapsilosis (24.1%), Candida tropicalis (13.2%), and Candida glabrata (12.8%). C. glabrata was relatively common in females (45.5%) compared to males. The age group 40-89 years was more frequently infected than other age groups, and the most frequent isolates according to age group were C. albicans in neonate (66.7%), C. parapsilosis in 1-9-year-olds (41.7%), and C. glabrata in those aged ≥60 years (range; 13.3%–20.0%). According to the visited departments, C. albicans, C. glabrata, and Candida haemulonii were more common in medical departments, while C. parapsilosis was more common in surgical departments. In the antifungal susceptibility test, a rising trend of azole resistance among C. albicans and C. glabrata was observed in recent years. CONCLUSION: In this study, it was confirmed that the isolation rate of Candida species in blood is different by age, gender, and hospital department, and the distribution of isolated Candida species changed over time. The resistance patterns of antifungal agents are also changing, and continuous monitoring and proper selection of antifungal agents are necessary.

Candida glabrata infection of urinary bladder in a Chinchilla Persian cat

A 5-year-old castrated male Chinchilla Persian cat weighing 4.84 kg was referred for hematuria. The cat had a history of urethrostomy and bacterial cystitis. In urine culture, Candida glabrata was cultured on Sabouraud dextrose agar. Based on these results, the cat was diagnosed with Candida cystitis. Subsequently, oral administration of fluconazole was initiated. Urine culture was negative at 31 days after administration. This case describes the diagnosis and treatment of Candida glabrata infection of urinary bladder in a cat with a history of urethrostomy.

Candida glabrata among Candida spp. from environmental health practitioners of a Brazilian Hospital

Abstract The incidence of the species Candida albicans and non-albicans Candida was evaluated in a Brazilian Tertiary Hospital from the environment and health practitioners. In a 12-month period we had a total positivity of 19.65% of Candida spp. The most recurring non-albicans Candida species was C. glabrata (37.62%), generally considered a species of low virulence, but with a higher mortality rate than C. albicans. Subsequently, C. parapsilosis (25.74%) and C. tropicalis (16.86%) were the second and third most commonly isolated species. Considering the total samples collected from the emergency room and from the inpatient and the pediatric sector, 19.10% were positive for Candida spp., with the predominance of non-albicans Candida species (89.42%). The high percentage of positivity occurred in the hands (24.32%) and the lab coats (21.88%) of the health care assistants. No sample of C. albicans presented a profile of resistance to the drugs. All the non-albicans Candida species presented a decreased susceptibility to miconazole and itraconazole, but they were susceptible to nystatin. Most of the isolates were susceptible to fluconazole and amphotericin B. As expected, a high resistance rate was observed in C. glabrata and C. krusei, which are intrinsically less susceptible to this antifungal agent. The contamination of environmental surfaces by Candida spp. through hand touching may facilitate the occurrence of Candida infections predominantly in immunocompromised patients. In addition to that, the antifungal agents used should be carefully evaluated considering local epidemiologic trends in Candida spp. infections, so that therapeutic choices may be better guided.