Research progress on drug resistance mechanism of Candida glabrata / 药学实践杂志

With the increasing proportion of Candida glabrata in patients with candidiasis, C. glabrata has become one of the most common pathogenic Candida in clinical practice. There are limited types of antifungal drugs, and the consequent problem of drug resistance is severely increasing, which brings difficulties to clinical treatment. The resistance mechanisms of C. glabrata to azoles, echinocandins and polyenes were reviewed in this paper.

Analysis of pathogens of mucosal candidiasis and their resistance to drugs in a third-grade class-A hospital in Taiyuan, Shanxi / 中华皮肤科杂志

Objective:To analyze types of mucosal candidiasis and drug resistance of relevant pathogens in a dermatology outpatient clinic in Taiyuan.Methods:Clinical data were collected from 172 patients with mucosal candidiasis, who had positive fungal culture results, in the dermatology outpatient clinic of Shanxi Bethune Hospital from 2019 to 2020. Pathogens were identified by a molecular biological approach, and in vitro drug sensitivity test was performed. Results:Among the 172 patients with mucosal candidiasis, 142 (82.6%) had vulvovaginal candidiasis, 24 (14.0%) had candidal balanoposthitis, and 6 (3.5%) had oral candidiasis; 3 patients were aged ≤ 18 years, 155 were aged 19 - 59 years, and 14 were aged ≥ 60 years, and the proportion of patients with vulvovaginal candidiasis significantly differed among the above 3 age groups (2/3, 134/155[86.45%], 6/14, respectively; χ2 = 14.29, P < 0.05) . Molecular biological identification showed that all the 172 isolated strains belonged to the genus Candida, including 165 strains of Candida albicans (95.9%) , 5 strains of Candida glabrata (2.9%) , and 2 strains of Candida parapsilosis (1.2%) ; the sensitivity to common antifungal agents including flucytosine, amphotericin B, fluconazole, itraconazole and voriconazole was 95.9%, 100.0%, 62.2%, 47.1% and 56.4%, respectively. Conclusion:In the dermatology outpatient clinic of Shanxi Bethune Hospital, vulvovaginal candidiasis was the most common type of mucosal candidiasis, and the main pathogen was Candida albicans; the Candida isolates showed high sensitivity to flucytosine and amphotericin B.

Emphysematous pyelonephritis caused by C. glabrata / Pielonefrite enfisematosa por C. glabrata

Abstract Emphysematous pyelonephritis (EPN) is a rare acute necrotizing infection of the kidney and surrounding tissues, with gas in the renal parenchyma, collecting system or perirenal tissue. The bacterial etiology predominates; mainly Gram-negative bacilli; Candida spp. and C. albicans are rarely described. We describe a case of EPN caused by C. glabrata, sensitive to fluconazole in a young, hypertensive woman with undiagnosed diabetes mellitus (DM), with renal dysfunction upon admission; her abdominal CT scan found a volumetric increase in the left kidney, signs of gas collections and perirenal blurring. Despite the antimicrobial therapy instituted, due to clinical refractoriness, a double J catheter and subsequent total nephrectomy were indicated, with good postoperative evolution. Her uroculture showed C. glabrata sensitive to fluconazole, and the pathology study showed tubular atrophy and intense interstitial inflammatory infiltrate. Despite the serious, potentially fatal condition, we could control the infection and the patient recovered fully. Poor DM management is an important triggering factor, and it is of great relevance to identify the EPN through imaging exams due to the peculiarities of its clinical and potentially surgical management

Resumo A pielonefrite enfisematosa (PNE) é uma infecção aguda rara necrotizante do rim e dos tecidos adjacentes, com presença de gás no parênquima renal, sistema coletor ou tecido perirrenal. Predomina a etiologia bacteriana, principalmente bacilos Gram-negativos; Candida spp. e na maioria das vezes C. albicans são raramente descritas. Descreve-se um caso de PNE causada por C. glabrata sensível a fluconazol em mulher jovem, hipertensa e com diabetes mellitus (DM) não diagnosticada, com disfunção renal à admissão; tomografia computadorizada de abdome constatou aumento volumétrico do rim esquerdo, sinais de coleções gasosas e borramento perirrenal. Apesar da terapia antimicrobiana instituída, devido à refratariedade clínica, foi indicado cateter duplo J e posterior nefrectomia total, com boa evolução pós-operatória. A urocultura evidenciou C. glabrata sensível a fluconazol, e o anatomopatológico demonstrou atrofia tubular e intenso infiltrado inflamatório intersticial. Apesar da condição grave, potencialmente fatal, houve controle do foco infeccioso e plena recuperação da paciente. O mau manejo do DM é um importante fator desencadeante, e é de grande relevância identificar a PNE por meio de exames de imagem devido às peculiaridades de seu manejo clínico e potencialmente cirúrgico.

Perfil de resistencia antifúngica en el tratamiento de candidiasis vaginal: Un diagnóstico de agentes etiológicos / Antifungal resistance profile in the treatment of vaginal candidiasis: A diagnosis of etiologic agents

Introducción: En las micosis, la resistencia a los antifúngicos aumenta debido a los diagnósticos y tratamientos incorrectos. Por tanto, se ha sugerido la vigilancia y el seguimiento de las cepas resistentes para garantizar una terapia adecuada. Objetivo: Determinar las especies de Candida y el perfil de resistencia a fluconazol y voriconazol, que presentan los aislados obtenidos de muestras almacenadas durante los meses diciembre 2017 y marzo 2018 de pacientes provenientes del Laboratorio del Hospital Regional "Miguel Ángel Mariscal Llerena" del departamento de Ayacucho, Perú. Material y Métodos: Estudio descriptivo transversal, en el cual se aislaron 110 cepas de Candida sp almacenadas por un período de cuatro meses, y procesadas por métodos estandarizados y aprobados por el Ministerio de Salud (MINSA) y el Instituto Nacional de Salud (INS) del Perú relacionados con el diagnóstico de agentes etiológicos de micosis humanas y la sensibilidad antifúngica con los métodos estandarizados de Clinical Laboratory Standard Institute (CLSI). Resultados: Se observó C. albicans en 86,4 % de los aislados seguida por C. glabrata con 9,1 %, C. parapsilosis 2,7 % y 0,9 % de C. tropicalis y C. krusei. El 10,5 % de C. albicans fue resistente al fluconazol y voriconazol con CMI ≥ 128 μg/mL y ≥ 16 μg/mL respectivamente, mientras que 20 % de C. glabrata mostró sensibilidad dosis dependiente y 10 % de resistencia al fluconazol. Conclusiones: Existe una gran variedad de especies de Candida, siendo la C. albicans la más común, seguida por C. glabrata, con un mayor porcentaje de aislamiento en comparación con otras especies. Se puede observar que estas especies poseen grados de vulnerabilidad considerables a la aplicación de fármacos como el fluconazol y voriconazo(AU)

Introduction: In mycoses, resistance to antifungals increases due to incorrect diagnosis and treatment. Therefore, surveillance and monitoring of resistant strains has been suggested to ensure adequate therapy. Objective: To determine the Candida species and the resistance profile to fluconazole and voriconazole presented by the isolates obtained from samples stored during the months of December 2017 and March 2018 from patients coming from the Laboratory of the Regional Hospital "Miguel Ángel Mariscal Llerena" in the department of Ayacucho, Peru. Material and Methods: Cross-sectional descriptive study in which 110 strains of Candida sp were isolated and stored for a period of four (04) months, and processed by standardized methods approved by the Ministry of Health (MINSA) and the National Institute of Health (INS) of Peru related to the diagnosis of etiological agents of human mycosis and antifungal sensitivity with the standardized methods of the Clinical Laboratory Standard Institute (CLSI). Results: C. albicans was observed in 86,4 % of isolates, followed by C. glabrata (9,1 %), C. parapsilosis (2,7 %), and 0,9 % of C. tropicalis and C. krusei; 10,5 % of C. albicans was resistant to fluconazole and voriconazole with MIC ≥ 128 μg/mL and ≥ 16 μg/mL respectively, while 20 % of C. glabrata showed dose dependent sensitivity and 10 % resistance to fluconazole. Conclusions: There is a wide variety of Candida species, with C. albicans being the most common, followed by C. glabrata, with a higher percentage of isolation compared to other species. It can be seen that these species have considerable degrees of vulnerability to the application of drugs such as fluconazole and voriconazole(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

Evaluation of in vitro antifungal effects of synthetic and herbal mouth rinses on oral Candida albicans and Candida glabrata

@#Mouth rinses which function as breath fresheners, medicaments, and antiseptics can also deliver oral therapeutic agents. This study evaluated and compared the antifungal effects of synthetic and herbal mouth rinses on oral C. albicans and C. glabrata via disk diffusion, minimal inhibition concentration (MIC), minimum fungicidal concentration (MFC), time-kill assay, and growth profile tests. The four chemical mouth rinses, namely Brand O (A), Brand M (B), Brand H (C), and Brand B (D) used in the study showed positive antifungal activity in these two species. The average diameter of the inhibition zones obtained from the disk diffusion test was higher in mouth rinse B (C. albicans = 12.0 ± 0.9 mm, C. glabrata = 13.5 ± 0.8 mm) compared to those in C, A and D. Both Candida species exhibited similar MIC and MFC values, ranging from 1.63 ± 0.5 to 18.75 ± 0.0 µg/mL and 6.51 ± 2.01 to 50.00 ± 9.36 µg/mL, respectively. These synthetic mouth rinses had efficient killing activity eliminating 50% of the growing population of both Candida spp. following 15 seconds exposure time. Analyses of the growth profile curves showed that mouth rinses B and A resulted in rapid growth depletion of both Candida spp. Meanwhile, three herbal mouth rinses, namely Brand S (E), Brand C (F), and Brand P (G), were less effective against C. albicans and C. glabrata. Mouth rinses B and A contained cetylpyridinium chloride and chlorhexidine, respectively, and could be an effective alternative for controlling and preventing oral candidiasis.

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)

Antifungal susceptibility profile of Candida clinical isolates from 22 hospitals of São Paulo State, Brazil

This study aimed to evaluate the frequency of cryptic Candida species from candidemia cases in 22 public hospitals in São Paulo State, Brazil, and their antifungal susceptibility profiles. During 2017 and 2018, 144 isolates were molecularly identified as 14 species; C. parapsilosis (32.6%), C. albicans (27.7%), C. tropicalis (14.6%), C. glabrata (9.7%), C. krusei (2.8%), C. orthopsilosis (2.8%), C. haemulonii var. vulnera (2.1%), C. haemulonii (1.4%), C. metapsilosis (1.4%), C. dubliniensis (1.4%), C. guilliermondii (1.4%), C. duobushaemulonii (0.7%), C. kefyr (0.7%), and C. pelliculosa (0.7%). Poor susceptibility to fluconazole was identified in 6.4% of C. parapsilosis isolates (0.12 to >64 µg/mL), 50% of C. guilliermondii (64 µg/mL), 66.6% of C. haemulonii var. vulnera (16-32 µg/mL), and C. duobushaemulonii strain (MIC 64 µg/mL). Our results corroborated the emergence of C. glabrata in Brazilian cases of candidemia as previously reported. Importantly, we observed a large proportion of non-wild type C. glabrata isolates to voriconazole (28.6%; <0.015 to 4 µg/mL) all of which were also resistant to fluconazole (28.6%). Of note, C. haemulonii, a multidrug resistant species, has emerged in the Southeast region of Brazil. Our findings suggested a possible epidemiologic change in the region with an increase in fluconazole-resistant species causing candidemia. We stress the relevance of routine accurate identification to properly manage therapy and monitor epidemiologic trends.

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.

Actinic cystitis associated with urinary tract infection by Candida glabrata as a differential diagnosis of pudendal neuralgia. Case report / Cistite actínica associada à infecção de trato urinário por Candida glabrata como diagnóstico diferencial de neuralgia do pudendo. Relato de caso

ABSTRACT BACKGROUND AND OBJECTIVES: Chronic pelvic pain is still a little-known syndrome with different etiological agents, high morbidity rate, with little information about its etiopathogenesis, which makes its treatment difficult, with symptoms that significantly impact the patient's quality of life. Among the conditions that lead to chronic pelvic pain, actinic cystitis stands out, a complication of pelvic radiotherapy. This pathology presents signs, symptoms, and complications similar to pudendal neuralgia. The objective of this report is to present one of the etiologies of chronic pelvic pain and its treatment. CASE REPORT: Sixty-three-year-old male patient who sought medical care with a history of chronic pelvic pain attributed to pudendal neuralgia. The diagnosis was actinic cystitis, resulting from pelvic radiotherapy for prostate adenocarcinoma and urinary tract infection by Candida glabrata, an opportunistic fungal agent. The patient was treated with fluconazole with total control of the painful symptoms and significant improvement in the quality of life. CONCLUSION: A case report with total pain control in a patient with an initial diagnosis of pudendal neuralgia, who after the clinical evaluation, imaging tests, cystoscopy, and lab tests was diagnosed with actinic cystitis associated to the urinary tract infection by Candida glabrata.

RESUMO JUSTIFICATIVA E OBJETIVOS: A dor pélvica crônica é uma síndrome ainda pouco conhecida, com diferentes agentes etiológicos, com alta morbidade, com poucas informações sobre sua etiopatogênese, dificultando seu tratamento, com sintomas que impactam de modo significativo a qualidade de vida do paciente. Entre as inúmeras causas está a cistite actínica, complicação da radioterapia pélvica, que apresenta sinais, sintomas e complicações semelhantes à neuralgia do pudendo. O objetivo deste relato foi apresentar uma das etiologias da dor pélvica crônica e seu tratamento. RELATO DO CASO: Paciente do sexo masculino, 63 anos, que procurou o serviço médico com história de dor pélvica crônica atribuída à neuralgia do pudendo. Foi feito o diagnóstico de cistite actínica causada por radioterapia pélvica para tratamento de adenocarcinoma de próstata e infecção de trato urinário pelo agente fúngico oportunista Candida glabrata, e instituído tratamento com fluconazol com controle total da sintomatologia dolorosa e melhora acentuada da qualidade de vida. CONCLUSÃO: Relato de caso com controle total da dor em paciente que apresentava diagnóstico inicial de neuralgia do pudendo, que após avaliação clínica, imaginológica, cistoscópica e laboratorial foi diagnosticado com cistite actínica associada à infecção de trato urinário por Candida glabrata.

Candida Species Causing Neonatal Septicaemia - Experience in a Tertiary Care Centre.

BACKGROUND Candida species is one of the most common causes of bloodstream infections among neonates and accounts for 9-13% of such infections. Non-albicans Candida have emerged as important opportunistic pathogens, notably C. tropicalis, C. glabrata, C. parapsilosis and C. krusei. With the introduction of antifungal agents, the causes of Candida infections shifted from an almost complete dominance of Candida albicans to the common involvement of Candida glabrata and other non-albicans Candida species.METHODSThis prospective, microbiological observational study was conducted in a tertiary care hospital for one and a half years from July 2016 to December 2017. Blood samples of neonates collected into Bactec Peds Plus/F Culture vials of an automated blood culture system (Bactec 9120, Becton Dikinson, USA) from clinically suspected cases of neonatal septicaemia were subjected to culture. Detailed clinical history such as presence of respiratory distress, abdominal distension, lethargy, feed intolerance, failure to thrive, poor perfusion, history of convulsions, duration of NICU stay and antibiotic use was taken from the medical records. The Candida species isolated were identified using standard mycological techniques. RESULTSOut of the total 250 clinically suspected cases of neonatal septicaemia, fungal growth was positive in 30 (12%) cases. Candida albicans accounted for 10% and non albicans Candida accounted for 90% of the fungal isolates. Candida glabrata (73.33%) was the commonest species followed by Candida tropicalis (16.67%) and Candida albicans (10%). Failure to thrive (60%), lethargy (50%) and respiratory distress (30%) were the most common clinical presentations seen, followed by feed intolerance (26.67%), abdominal distension (13.33%) and fever (10%). Among the risk factors observed for neonatal candidemia, low birth weight (76.67%) and prematurity (73.33%) were commonest followed by broad spectrum antibiotic use (66.67%), total parenteral nutrition (53.3%), ventilator support (36.67%) and indwelling catheters (26.67%).CONCLUSIONSNon-albicans Candida has emerged as an important pathogen causing neonatal septicaemia. Fungal sepsis is more common among preterm, low birth weight infants, those with prolonged antibiotic use and those on total parenteral nutrition. Candida glabrata caused the highest number of cases of fungal septicaemia

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.

Multidrug-resistant Candida glabrata strains obtained by induction of anidulafungin resistance in planktonic and biofilm cells

Candida glabrata has emerged as a common cause of serious life-threatening fungal infections, largely owing to their low susceptibility to azole antifungals. Recent guidance indicates the use of echinocandins as the first-choice drug for the treatment of systemic infections of C. glabrata; however, C. glabrata resistance to echinocandins is reportedly increasing. Herein, we present the induction of anidulafungin resistance in planktonic and sessile cells of C. glabrata and the development of fluconazole cross-resistance. MICs of 21 clinical C. glabrata strains were determined by a broth microdilution method using anidulafungin and fluconazole. Biofilm formation on a tracheal catheter was determined using 1- × 1-cm2 polyvinyl polychloride catheter fragments. Induction of anidulafungin resistance in planktonic and sessile cells and evaluation of its stability were performed by exposing the strains to successively higher concentrations of the antifungal. The induction resulted in strains strongly resistant to anidulafungin (MICs: 1-2 µg/mL) and fluconazole (≥64 µg/mL). Most of the sessile cells of C. glabrata presented slightly reduced susceptibility compared with the planktonic cells. Clinically, this cross-resistance could lead to therapeutic failure while using fluconazole in patients previously exposed to subinhibitory concentrations of anidulafungin for extended periods.

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.

Changing virulence factors among vaginal non-albicans Candida species

Background: Vulvovaginal candidiasis (VVC) is caused by overgrowth of Candida species in the female lower genital tract and most commonly caused by Candida albicans. The production of various virulence factors may attribute to their pathogenicity. Hence, this study was aimed to determine the production of various virulence factors of Candida spp. causing VVC. Materials and Methods: A total of 51 Candida spp. were isolated prospectively from 50 patients among 211 clinically suspected cases of VVC. The haemolytic activity, biofilm production, proteinase activity, phospholipase activity and esterase activity were detected by standard methods. Statistical analysis was performed using OpenEpi version 3.01. Results: Haemolytic activity was observed in 42 Candida isolates (82.4%), biofilm activity in 21 Candida isolates (41.2%), proteinase and esterase activity in 19 Candida isolates (37.3%) each and phospholipase activity in 15 Candida isolates (29.4%). Phospholipase activity was observed in all of the C. albicans strains, whereas all strains of Candida krusei were able to produce biofilm. All strains of Candida parapsilosis and 87% strains of Candida glabrata were haemolytic. Five of the eight C. glabrata strains were found to produce strong proteinase (Prz score ?0.63). About 30.4% strains of C. glabrata and 20% strains of C. krusei were found to be positive for esterase activity. This is one of the few studies which revealed esterase activity among C. glabrata and C. krusei strains. Conclusions: This study highlighted that there is a change in the virulence factors among the non-albicans Candida species, especially C. glabrata strains which were haemolytic and produce strong proteinase activity and esterase activity. It may be one of the explanation of the most common causative agent of VVC in our study. Multicentric studies from this area might be required to get a more generalised conclusion.

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.

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.

Gene polymorphism and drug sensitivity of Candida glabrata associated with vulvovaginal candidiasis in women of childbearing age / 中华检验医学杂志

Objective@#To investigate genotypes and drug susceptibility of the 100 strains of Candida glabrata isolated from 100 women (including 50 pregnant women) in order to study the drug-resistance and gene polymorphism, and to investigate the correlation of drug-resistance, gestation and gene polymorphism.@*Methods@#Multilocus sequence typing (MLST) technique was introduced to identify sequences of 6 housekeeping genes from 100 isolates of Candida glabrata. The results were compared with sequence information in MLST databases by Clustalx software to determine a strain allelic profile and sequence type (ST). Drawing the phylogenetic tree by weighted paired group average method and the minimal spanning tree method of MEGA6.0 software, the microevolution and relationship between different strains were analyzed. ATB FUNGUS semi-automatic system was used to test the drug susceptibility. Fisher analysis method was used to analyze the correlation between the genotypes and pregnancy.Ridit analysis method was used to analyze the correlation between the genotypes and drug susceptibility.@*Results@#The 100 isolates belonged to 34 clone sequences. There were 53 isolates belonged to ST-7 ( 26 isolates of pregnancy), 7 isolates belonged to ST-3 (3 isolates of pregnancy), 6 isolates belonged to ST-19 (5 isolates of pregnancy ), 3 isolates belonged to ST-15 (1 isolate of pregnancy), 2 isolates belonged to ST-10 (2 isolates of pregnancy), 1 isolate belonged to the other types of ST. Total of 100 isolates of Candida glabrata were 100% sensitive to fluorocytosine and amphotericin. The effect of itraconazole was poor with the sensitive rate of 20%. The resistance rates of fluconazole and voriconazole were 4% and 1% respectively. All genotypes were sensitive to voriconazole except ST-X1. In the correlation between genotype and itraconazole resistance, ST-7 as the standard group, the Ridit values in the group of ST-15, ST-19 and other types of ST were not included the mean Ridit value of the standard group in itraconazole (0.5). The system evolution tree was built using the neighbor-joining method (NJ) . All genotypes could be divided into 3 groups, as Ⅰ, Ⅱ, Ⅲ. Group Ⅰ had 44 cases, group Ⅱ had 53 cases , group Ⅲ had 3 cases . All the collected clinical strains had small genetic distances during molecular evolution.@*Conclusions@#ST-7 was the dominant genotype in Guiyang. No correlation between different STs and patients′ pregnancy was found. The different drug susceptibility in itraconazole between ST-7, ST-15, ST-19 and other STs were found. The Candida glabrata associated with VVC showed highly discrimative diversity. However, the phylogenic analysis exhibited genetic similarity among the strains studied.(Chin J Lab Med, 2018, 41: 596-600)

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.

Study of Microbial Interaction Formed by "Candida krusei" and "Candida glabrata": "In Vitro" and "In Vivo" Studies

Abstract Recently, the non-albicans Candida species have become recognized as an important source of infection and oral colonization by association of different species in a large number of immunosuppressed patients. The objective of this study was to evaluate the interactions between C. krusei and C. glabrata in biofilms formed in vitro and their ability to colonize the oral cavity of mouse model. Monospecies and mixed biofilms were developed of each strain, on 96-well microtiter plates for 48 h. These biofilms were analyzed by counting colony-forming units (CFU/mL) and by determining cell viability, using the XTT hydroxide colorimetric assay. For the in vivo study, twenty-four mice received topical applications of monospecie or mixed suspensions of each strain. After 48 h, yeasts were recovered from the mice and quantified by CFU/mL count. In the biofilm assays, the results for the CFU/mL count and the XTT assay showed that the two species studied were capable of forming high levels of in vitro monospecie biofilm. In mixed biofilm, the CFU of C. krusei increased (p=0.0001) and C. glabrata decreased (p=0.0001). The metabolic activity observed in XTT assay of mixed biofilm was significantly reduced compared with a single C. glabrata biofilm (p=0.0001). Agreeing with CFU in vitro count, C. glabrata CFU/mL values recovered from oral cavity of mice were statistically higher in the group with single infection (p=0.0001) than the group with mixed infection. We concluded that C. krusei inhibits C. glabrata and takes advantage to colonize the oral cavity and to form biofilms.

Resumo Recentemente, as espécies não albicans tem se tornado uma importante fonte de infecção e de colonização oral pela associação de espécies em um grande número de pacientes imunossuprimidos. O objetivo desse estudo foi avaliar a interação entre C. krusei e C. glabrata em biofilmes formados in vitro e sua capacidade em colonizar a cavidade oral em modelo de camundongo. Biofilmes monoespécies e mistos foram formados em placas de 96 poços por 48 h. Esses biofilmes foram analisados pela contagem de UFC/mL e pela determinação da viabilidade celular, usando ensaio de XTT. Para o estudo in vivo, vinte e quatro camundongos receberam aplicações tópicas de suspensões monoespécies e mistas de cada espécie. Após 48 h, as leveduras foram recuperadas dos camundongos e quantificadas por UFC/mL. Nos ensaios de biofilme, os resultados da contagem de UFC/mL e do ensaio de XTT mostraram que as duas espécies estudadas foram capazes de formar grande quantidade de biofilme monoespécie in vitro. Nos biofilmes mistos, a UFC/mL de C. krusei aumentou (p=0,0001) e de C. glabrata diminuiu (p=0,0001). A atividade metabólica observada no ensaio de XTT nos biofilmes mistos foi significantemente reduzida comparada com o biofilme formado apenas de C. glabrata (p=0,0001). Concordado com as contagens in vitro, os valores de UFC/mL de C. glabrata recuperados da cavidade oral dos camundongos foram estatisticamente maior no grupo com infecção simples (p=0,0001) do que do grupo com infecção mista. Nós concluímos que C. krusei inibe C. glabrata e possui vantagem em colonizar a cavidade oral e formar biofilmes.