Identificación de cepas del complejo Candida albicans aisladas de muestras clínicas en la región de Valparaíso, Chile / Identification of strains of the Candida albicans complex isolated clinical samples in the Valparaíso region, Chile

Introducción: C. albicans es reconocida como la especie más virulenta del género y representa la causa más frecuente de candidiasis en humanos. A nivel taxonómico, C.albicans se clasifica como un complejo de especies estrechamente relacionadas que incluye a C. albicans sensu stricto (s.s), C. dubliniensis y C. africana. Objetivo: identificar las especies del complejo C. albicans aisladas desde distintas muestras de pacientes de la quinta región de Valparaíso. Materiales y método: Se identificaron 103 cepas del complejo C. albicans, aisladas desde muestras superficiales y profundas durante el año 2020. La identificación se realizó en base a morfofisiología y la amplificación del gen HWP1. Resultados: Se identificaron 100 cepas como C. albicans s.s, 2 como C. dubliniensis y 1 como C. africana. Dentro de las cepas identificadas como C. albicans s.s se observaron cuatro patrones de tamaños de fragmentos genéticos. Conclusiones: C. albicans s.s fue la especie más frecuente y en base al genotipo de HPW1 se describen cuatro patrones ( H1 a H4). (AU)

Introduction: C. albicans is recognized as the most virulent species of the genus and represents the major cause of candidiasis in humans. At the taxonomic level, C. albicansis classified as a complex of closely related species that includes C. albicans sensu stricto (s.s), C. dubliniensis, and C. africana. Objective: to identify the species of the C. albicans complex isolated from different samples of patients from the fifth region of Valparaíso. Materials and method: 103 strains of the C. albicans complex were identified, isolated from superficial and deep samples during the year 2020. The identification was carried out based on morphophysiology and the amplification of the HWP1 gene. Results: 100 strains were identified as C. albicans s.s, 2 as C. dubliniensis and 1 as C. africana. Within the strains identified as C. albicans s.s, 4 patterns of fragment sizes were observed. Conclusions: C. albicans s.s was the most frequent species and based on the HPW1 genotype, four patterns are described (H1 to H4).(AU)

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

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

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

Características de las fungemias en un centro de referencia del Perú: retrospectiva de cinco años / Characteristics of fungemia in a peruvian referral center: 5-year retrospective analysis

RESUMEN Con el objetivo de conocer las características de las fungemias en 285 pacientes oncológicos hospitalizados del 2012 al 2016 en el Instituto Nacional de Enfermedades Neoplásicas se realizó un estudio descriptivo retrospectivo. Se evaluó información demográfica, clínica y microbiológica. Las fungemias por C. albicans predominaron en pacientes con tumores sólidos y sin neutropenia, mientras las causadas por C. tropicalis predominaron en pacientes con neoplasias hematológicas y neutropenia. C. tropicalis fue el agente más aislado (47,0%). Las fungemias aumentaron con el tiempo en los pacientes sin neutropenia. Las fungemias causadas por C. albicans aumentan con la edad en pacientes con tumores sólidos sin neutropenia. Se concluye que las fungemias son mayormente causadas por C. tropicalis en pacientes con neoplasias hematológicas con neutropenia y por C. albicans en pacientes con tumores sólidos sin neutropenia. Además, las fungemias en pacientes sin neutropenia aumentan en el tiempo y las causadas por C. albicans, en tumores sólidos sin neutropenia, aumentan con la edad.

ABSTRACT Retrospective descriptive study carried out to determine the characteristics of fungemia in 285 cancer patients hospitalized from 2012 to 2016 at the Instituto Nacional de Enfermedades Neoplásicas (INEN). Demographic, clinical and microbiological information was evaluated. Fungemia by C. albicans predominated in patients with solid tumors and without neutropenia, while those caused by C. tropicalis predominated in patients with hematological neoplasia and neutropenia. C. tropicalis was the agent isolated in most cases (47.0%). Fungemia increased over time in patients without neutropenia. Fungemia caused by C. albicans increases with age in patients with solid tumors without neutropenia. It is concluded that fungemia are mainly caused by C. tropicalis in patients with hematological neoplasia with neutropenia and by C. albicans in patients with solid tumors without neutropenia. In addition, fungemia in patients without neutropenia increases over time; and those caused by C. albicans increase with age in patients with solid tumors without neutropenia.

The role of Candida albicans in root caries biofilms: an RNA-seq analysis

Abstract Objective This study sought to analyze the gene expression of Candida albicans in sound root surface and root caries lesions, exploring its role in root caries pathogenesis. Methodology The differential gene expression of C. albicans and the specific genes related to cariogenic traits were studied in association with samples of biofilm collected from exposed sound root surface (SRS, n=10) and from biofilm and carious dentin of active root carious lesions (RC, n=9). The total microbial RNA was extracted, and the cDNA libraries were prepared and sequenced on the Illumina Hi-Seq2500. Unique reads were mapped to 163 oral microbial reference genomes including two chromosomes of C. albicans SC5314 (14,217 genes). The putative presence of C. albicans was estimated (sum of reads/total number of genes≥1) in each sample. Count data were normalized (using the DESeq method package) to analyze differential gene expression (using the DESeq2R package) applying the Benjamini-Hochberg correction (FDR<0.05). Results Two genes (CaO19.610, FDR=0.009; CaO19.2506, FDR=0.018) were up-regulated on SRS, and their functions are related to biofilm formation. Seven genes ( UTP20 , FDR=0.018; ITR1 , FDR=0.036; DHN6 , FDR=0.046; CaO19.7197 , FDR=0.046; CaO19.7838 , FDR=0.046; STT4 , FDR=0.046; GUT1 , FDR=0.046) were up-regulated on RC and their functions are related to metabolic activity, sugar transport, stress tolerance, invasion and pH regulation. The use of alternative carbon sources, including lactate, and the ability to form hypha may be a unique trait of C. albicans influencing biofilm virulence. Conclusions C. albicans is metabolically active in SRS and RC biofilm, with different roles in health and disease.

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

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

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

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

Factores relacionados a candidiasis oral en niños y adolescentes con VIH, caracterización de especies y susceptibilidad antifúngica / Factors related to oral candidiasis in HIV children and adolescents, species characterization and antifungal susceptibility

Resumen Introducción: Se desconocen los factores asociados a la candidiasis oral en población pediátrica con infección por VIH de los países en desarrollo. Objetivo: Identificar los factores asociados a la colonización por Candida, candidiasis oral y la susceptibilidad in vitro a antifúngicos, en niños y adolescentes con infección por VIH institucionalizados en la ciudad de Tijuana, México. Materiales y Métodos: Se examinó la cavidad oral de 30 niños y adolescentes con infección por VIH, se obtuvo una muestra de la mucosa oral para identificar las especies de Candida mediante cultivo y auxonograma. La susceptibilidad a los antifúngicos se determinó de acuerdo al CLSI. Los indicadores del estado inmunológico y falla virológica se clasificaron conforme a la OMS. Resultados: Se identificaron seis especies de Candida, 53% colonizantes y 47% causantes de candidiasis. Los factores asociados a candidiasis fueron alta carga viral (p = 0,001), menor recuento de LTCD4+ (p = 0,002) y esquema TARAA (p ≤ 0,014). La especie prevalente fue C. glabrata (33%); sin embargo, C. albicans (27%) fue más resistente a fluconazol (p = 0,001). Las especies resistentes a itraconazol se identificaron en esquemas que incluyen un INNTR (p = 0,041). Conclusiones: Los niños y adolescentes con infección por VIH institucionalizados mostraron una prevalencia elevada de Candida spp. colonizante y resistencia a los antifúngicos relacionada con los INNTR .

Background: Factors associated with candidiasis and colonization in HIV-positive children and adolescents in developing countries are not well understood. Aim: To identify the factors associated with oral Candida colonization and candidiasis in institutionalized HIV-positive children and adolescents in Tijuana, México, as well as the response of the isolates to antifungals. Materials and Methods: Sample of the oral mucosa of 30 HIV positive children and adolescents were obtained to isolate and identify Candida species by culture and metabolic profile. Antifungal drugs susceptibility was determined according to CLSI. Indicators of immunological and virologic failure were classified in accordance to WHO criteria. Results: Six Candida species were identified from oral mucosa, 53% colonizers and 47% in candidiasis. Factors associated with candidiasis and oral colonization were viral load (p = 0,001), CD4+ counts (p = 0,002) and HAART regimen (p ≤ 0,014). The most prevalent species was C. glabrata (33%), but C. albicans (27%) was more resistant to fluconazole (p = 0,001). Itraconazol resistant species were identified in regimens that include an NNRTI (p = 0,041). Conclusion: HIV-positive children and adolescents living in an orphanage showed high prevalence of colonizing Candida spp. and resistance to antifungals, related to NNRTI.

Comparison of the recovery rate of otomycosis using betadine and clotrimazole topical treatment / Comparação da taxa de recuperação de otomicose com o uso de tratamento tópico com betadina e clotrimazol

Abstract Introduction Otomycosis is a common diseases that can be associated with many complications including involvement of the inner ear and mortality in rare cases. Management of otomycosis can be challenging, and requires a close follow-up. Treatment options for otomycosis include local debridement, local and systemic antifungal agents and utilization of topical antiseptics. Objective This study was designed to compare the recovery rate of otomycosis using two therapeutic methods; topical betadine (Povidone-iodine) and clotrimazole. Methods In this single-blind clinical trial, 204 patients with otomycosis were selected using a non-probability convenient sampling method and were randomly assigned to two treatment groups of topical betadine and clotrimazole (102 patients in each group). Response to treatment was assessed at 4, 10 and 20 days after treatment. Data were analyzed using the independent t-test, Chi-Square and Fisher exact test in SPSS v.18 software, at a significance level of p < 0.05. Results The results showed that out of 204 patients with otomycosis, fungi type isolated included Aspergillus in 151 cases (74%), and Candida albicans in 53 patients (26%). On the fourth day after treatment, 13 patients (13.1%) in the group treated with betadine and 10 patients (9.8%) in the group treated with clotrimazole showed a good clinical response to treatment (p = 0.75). A good response to treatment was reported for 44 (43.1%) and 47 patients (46.1%) on the tenth day after the treatment (p = 0.85); and 70 (68.6%) and 68 patients (67.6%) on the twentieth day after treatment (p = 0.46) in the groups treated with betadine and clotrimazole, respectively. The response to treatment was thus not significantly different in the two groups. Conclusion In the present study the efficacy of betadine and clotrimazole was the same for the treatment of otomycosis. The result of this study supports the use of betadine as an effective antifungal in otomycosis treatment, helping to avoid the emergence of resistant organisms.

Resumo Introdução A otomicose é uma das doenças comuns associadas a muitas complicações, como envolvimento da orelha interna e mortalidade em casos raros. O tratamento da otomicose pode ser realmente desafiador e requer um acompanhamento rigoroso. As opções de tratamento para otomicose podem incluir desbridamento local, agentes antifúngicos locais e sistêmicos e uso de antissépticos tópicos, os medicamentos tópicos recomendados para o tratamento da otomicose. Objetivo Comparar a taxa de recuperação de otomicose utilizando dois métodos terapêuticos de betadina tópica (povidona-iodo) e clotrimazol. Método Neste ensaio clínico simples cego, 204 pacientes com otomicose foram selecionados utilizando-se método de amostragem de não probabilidade conveniente e randomizados para dois grupos de tratamento, com betadina tópica e com clotrimazol (102 pacientes em cada grupo). A resposta ao tratamento foi avaliada aos 4, 10 e 20 dias após o tratamento. Os dados foram analisados utilizando o teste t independente, qui-quadrado e teste de Fisher no software SPSS v.18, com nível de significância de p < 0,05. Resultados Os resultados mostraram que dos 204 pacientes com otomicose, os tipos de fungos isolados incluíram Aspergillus em 151 casos (74%) e Candida albicans em 53 pacientes (26%). No quarto dia após o tratamento, 13 pacientes (13,1%) no grupo tratado com betadina e 10 pacientes (9,8%) no grupo tratado com clotrimazol apresentaram boa resposta ao tratamento (p = 0,75). Uma boa resposta ao tratamento foi relatada para 44 (43,1%) e 47 pacientes (46,1%) no décimo dia após o tratamento (p = 0,85); e 70 (68,6%) e 68 pacientes (67,6%) no vigésimo dia após o tratamento (p = 0,46) no grupo tratado com betadina e clotrimazol, respectivamente. Assim, a resposta ao tratamento não foi significativamente diferente nos dois grupos. Conclusão No presente estudo, a eficácia da betadina e do clotrimazol foi a mesma no tratamento da otomicose. O resultado deste estudo apoia o uso de betadina como um antifúngico eficaz no tratamento da otomicose que pode ajudar a evitar o surgimento de organismos resistentes.

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.

Determination of the effective anticandidal concentration of denture cleanser tablets on some denture base resins

Abstract Objective Although the effectiveness of chemical cleansing against Candida albicans biofilm has been shown, the effective concentration of denture cleanser tablets has not been studied. The aim of this study was to assess the effect of three denture materials against Candida albicans biofilm and to determine effective concentrations of denture cleanser tablets. Material and methods The surface-roughness of Acron-hi™, QC-20™ and Deflex™ (n=45 per resin) resins was standardized by using a profilometer and their contact angle or surface free energy was calculated. C. albicans biofilm was formed on all three resins and were treated with Polident 3 min™, Corega™ and Fittydent™ cleanser solutions at various concentrations and both resin-biofilm and cleanser-biofilm interest were determined by using a MTT protocol according to the European Committee on Antimicrobial Susceptibility Testing's antifungal susceptibility testing (AFST-EUCAST). Scanning electron microscopy was used to compare the efficacy of different resin materials against C. albicans biofilm. Anticandidal activity and surface free energy statistical parameters were calculated by using 3-way and 1-way ANOVA, respectively (p<0.05). Results Polident 3 min™ and Corega™ tablets significantly inhibited (p<0.05) the proliferation of C. albicans against all denture resins at 27-37 mg/mL. Scanning electron microscopy results indicated that there was no significant difference among resin specimens regarding biofilm formation on dentures. We failed to find a significant relationship between surface free energy and the anticandidal effect of resin types. However, the polarity value of the resins was statistically associated with their anticandidal activity. Conclusions The polarity of the resins, the concentrations of tablets and the chemical content of the cleanser may directly affect C. albicans biofilm formations. Polident 3 min™ and Corega™ tablets should be suggested for patients who use any denture resin types, whereas the Fittydent™ tablet should only be proposed for those who use Deflex™, when two tablets are dropped into 150 mL water.

Phagocytosis and nitric oxide production by peritoneal adherent cells in response to Candida albicans in aging: a collaboration to elucidate the pathogenesis of denture stomatitis

Abstract Elderly denture wearers are commonly affected by Candida-associated denture stomatitis (DS), an inflammatory process of the oral mucosa strongly associated with Candida spp and other microorganisms, as well as local and systemic factors. The impaired immune response against pathogens is among the inherent host factors that have been also associated with the pathogenesis of DS. Mononuclear phagocytes respond to the pathogens through phagocytosis followed by the production of several substances inside the phagosomes, among them are the reactive nitrogen species (RNS). A failure in these mechanisms may contribute to the DS development. Objective The aim of this study was to investigate the influence of aging on the internalization and the production of nitric oxide (NO) by peritoneal adherent cells (PAC), in response to Candida albicans (C. albicans). Material and methods PAC obtained from young and aged mice were challenged with dead or viable C. albicans by using predetermined proportions (cells:yeast) for 30 and 120 minutes. Phagocytosis was analyzed by acridine orange dye, and NO production by the Griess reaction. Results C. albicans phagocytosis by PAC from aged mice was similar to that of young mice, although the cells from older mice cells present more internalized fungi compared with matched control. In addition, a tendency towards impaired NO production by peritoneal mononuclear phagocytes from aged mice was observed. Conclusions PAC from aged mice may capture and store many fungi, which in turn may mean that these cells are effectively unable to eliminate fungi, probably due to impaired NO production. Therefore, considering the important role of C. albicans overgrowth in the pathogenesis of DS and the aspects observed in this study, aging may favor the onset and severity of local candidosis such as DS and its systemic forms.

Investigations of the prevalence and virulence of Candida albicans in periodontal and endodontic lesions in diabetic and normoglycemic patients

Abstract Pulpal and periodontal tissues have similar microbiota that allows cross-contamination between the pulp and periodontal tissues. Objective The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. Material and Methods A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. Results C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). Conclusions Candida albicans was isolated more frequently and had higher virulence in diabetic patients.

Frecuencia de candidiasis oral asociada al uso de prótesis dentales en pacientes de la clínica odontológica de la Universidad Anáhuac Norte / The frequency of oral candidiasis associated with the use of dental prostheses in patients of the dental clinic at the Anahuac University

La candidiasis es la infección micótica más común de la cavidad bucaly es causada por el hongo Candida. Dentro de la población geriátricala candidiasis oral es uno de los tres principales motivos de consulta. El crecimiento en superfi cies es parte natural del modo de vivir del hongo Candida y es común que colonice las prótesis dentales, dando como resultado estomatitis por uso de dentadura o estomatitis subplaca.El diagnóstico de estomatitis por dentadura es relevante, ya quealrededor de 50 por ciento de las personas de edad promedio de 65 a 74 años, y 70 por ciento de 75 a 84 años utilizan prótesis dentales removibles. El objetivo del presente estudio fue determinar la frecuencia de candidiasis oral asociada a prótesis dentales removibles en los pacientes de la ClínicaOdontológica de la Universidad Anáhuac México Norte durante el periodo enero-mayo de 2016. Material y métodos: Se realizó un estudiodescriptivo, transversal y observacional. La muestra estuvo conformada por pacientes portadores de prótesis dentales removibles que acudieron a la Clínica Odontológica de la Universidad Anáhuac México Norte.Se interrogó al paciente respecto al uso de la prótesis y su condición de salud, posterior a la exploración clínica se tomó la citología exfoliativa de la mucosa debajo de la prótesis removible. Resultados: La muestraestuvo constituida por 22 pacientes de la clínica de prótesis removible,seis eran portadores de prótesis, de éstos 83 por ciento fueron positivos a candidiasis. Conclusiones: La falta de indicaciones sobre el cuidado, uso ehigiene de las prótesis dentales o la falta de apego a estas indicacionespor parte del paciente ocasionan el desarrollo de estomatitis subplaca.

Candidiasis is the most common mycotic infection in oral cavity andcaused by fungi candid. Oral candidiasis is one of three principalreasons for consultation in a geriatric population. A natural part of theway of living of the fungi Candida is the growth on surfaces. It growthmakes common that the fungi Candida colonize the dental prostheses,so the denture stomatitis or dental-related stomatitis is development.Denture stomatitis diagnosis is quite important because from the totalpopulation using dental removable prostheses around of 50% is between65-74 years old and 70% is between 75-84 years old. The ofthis study was identifying oral candidiasis frequency related to dentalremovable prostheses in patients from Universidad Anahuac, DentalSchool (México Norte), from January 2016 to May 2016. Material andmethods: Was made a descriptive, cross and observational study. Thesample was integrated by patients using dental removable prostheses,who went to Universidad Anahuac’s dental school (México Norte). Thepatients were interrogated about dental removable prostheses use andtheir health condition, after clinic exploration and exfoliative cytologywas taken from mucosa below dental removable prostheses. Results: 22patients from dental service using removable prosthesis constituted thesample, from this 22 patients only 6 were using a removable prosthesisand from this 6 patients only 83% were positive to oral candidiasis.Conclusions: Instructions scare about care, use, and hygiene of dentalprosthesis as well as not follow these instructions for patient’s partmake possible denture stomatitis development.

Association between pathogens from tracheal aspirate and oral biofilm of patients on mechanical ventilation

Abstract The aim of this study was to detect possible associations between respiratory pathogens from tracheal aspirate and oral biofilm samples in intubated patients in an intensive care unit (ICU), and to identify the most common respiratory pathogens in oral biofilm, particularly in patients that developed ventilator-associated pneumonia (VAP). Two oral biofilm samples were collected from the tongue of intubated patients (at admission and after 48 hours) and analyzed by culture with the Antibiotic Sensitivity Test. The results from the tongue biofilm samples were compared with the tracheal secretions samples. A total of 59.37% of patients exhibited the same species of pathogens in their tracheal aspirate and oral biofilm, of which 8 (42.1%) developed VAP, 10 (52.63%) did not develop pneumonia and one (5.26%) had aspiration pneumonia. There was a statistically significant association between presence of microorganisms in the tracheal and mouth samples for the following pathogens: Klebsiella pneumoniae, Candida albicans, Pseudomonas aeruginosa, Enterobacter gergoviae, Streptococcus spp and Serratia marcescens (p < 0.05). Pathogens that are present in tracheal aspirates of intubated patients can be detected in their oral cavity, especially in those who developed VAP or aspiration pneumonia. Thus, the results indicate that an improved oral care in these patients could decrease ICU pneumonia rates.

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

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

Phenotypic and genotypic detection of Candida albicans and Candida dubliniensis strains isolated from oral mucosa of AIDS pediatric patients

ABSTRACT The aim of this study was to assess a collection of yeasts to verify the presence of Candida dubliniensis among strains isolated from the oral mucosa of AIDS pediatric patients which were initially characterized as Candida albicans by the traditional phenotypic method, as well as to evaluate the main phenotypic methods used in the discrimination between the two species and confirm the identification through genotypic techniques, i.e., DNA sequencing. Twenty-nine samples of C. albicans isolated from this population and kept in a fungi collection were evaluated and re-characterized. In order to differentiate the two species, phenotypic tests (Thermotolerance tests, Chromogenic medium, Staib agar, Tobacco agar, Hypertonic medium) were performed and genotypic techniques using DNA sequencing were employed for confirmation of isolated species. Susceptibility and specificity were calculated for each test. No phenotypic test alone was sufficient to provide definitive identification of C. dubliniensis or C. albicans, as opposed to results of molecular tests. After amplification and sequencing of specific regions of the 29 studied strains, 93.1% of the isolates were identified as C. albicans and 6.9% as C. dubliniensis. The Staib agar assay showed a higher susceptibility (96.3%) in comparison with other phenotypic techniques. Therefore, genotypic methods are indispensable for the conclusive identification and differentiation between these species.

1, 3, 4-Thiadiazole and 1, 2, 4-triazole-3(4 H )-thione bearing salicylate moiety: synthesis and evaluation as anti- Candida albicans

Abstract Dramatically increased occurrence of both superficial and invasive fungal infections has been observed. Candida albicans appear to be the main etiological agent of invasive fungal infections. The anti-C. albicans activity of thiosemicarbazide, 1,3,4-Thiadiazole, and 1,2,4-triazole-3(4H)-thione compounds (compounds 3-23) were investigated. The MIC values of thiadiazole and triazole derivatives 10-23 were in the range of 0.08-0.17 µmol mL-1, while that of fluconazole was 0.052 µmol mL-1. Compound 11 (5-(2-(4-chlorobenzyloxy)phenyl)-N-allyl-1,3,4-thiadiazol-2-amine) and compound 18 (5-(2-(4-chlorobenzyloxy)phenyl)-4-allyl-2H-1,2,4-triazole-3(4H)-thione) were found to be the most active compounds, with MIC values of 0.08 µmol mL-1. The newly synthesized thiadiazole and triazole compounds (compounds 10-23) showed promising anti-Candida activity. The allyl substituent-bearing compounds 11 and 18 exhibited significant anti-Candida albicans activity and showed a binding mode as well as the fluconazole x-ray structure.

Síntese de heteroaril oxazolinas e derivados triazólicos com potencial atividade biológica / Synthesis of heteroaryl oxazolines and triazolic derivatives with potential biological activity

Fungos no ambiente podem ser patogênicos ou oportunistas, dependendo da imunidade do hospedeiro. Existem várias espécies de fungos, por exemplo, Cândida albicans, Cryptococcus e Aspergillus. A primeira espécie fúngica pode ser tratada com o antifúngico fluconazol, que é um composto que contém anéis heterocíclicos 1,2,4-triazólicos. Além disso, existem cepas de fungos que são resistentes à terapia com fluconazol, que é o caso das Cândida krusei, Cândida tropicalis; entre outras. A busca por novos tratamentos envolve o desenvolvimento de novas moléculas sintéticas. Neste trabalho, sintetizamos uma biblioteca de compostos oxazolínicos e seus derivados 1,2,3-triazólicos. A atividade microbiológica foi avaliada contra 10 tipos de Cândida, 2 tipos de Cryptococcus e 2 tipos de Aspergillus. Além disso, foram feitos os testes de hemólise, citotoxicidade, combinações de drogas e permeabilidade de membrana. Os resultados sugerem um alto potencial terapêutico dos compostos e os propomos como potenciais novos antifúngicos

ungi in the environment may be pathogenic or opportunistic depending on the immune status of the host. There are several species of fungi, for example, Candida albicans, Cryptococcus and Aspergillus. The first fungal species can be treated with the antifungal fluconazole, which is a compound containing 1,2,4-triazole heterocyclic rings. In addition, there are strains of fungi that are resistant to fluconazole therapy, which is the case of Candida krusei, Candida tropicalis; among others. The search for new treatments involves the development of new synthetic molecules. In this work, we synthesized a library of oxazoline compounds and their 1,2,3-triazole derivatives. Microbiological activity was evaluated against 10 types of Candida, 2 types of Cryptococcus and 2 types of Aspergillus. In addition, hemolysis, cytotoxicity, drug combinations and membrane permeability were performed. The results suggest the high therapeutic potential of the compounds and we propose them as potential new antifungals

Extracellular phospholipase production of oral Candida albicans isolates from smokers, diabetics, asthmatics, denture wearers and healthy individuals following brief exposure to polyene, echinocandin and azole antimycotics

Abstract Objective Candida albicans is the primary causative agent of oral candidosis, and one of its key virulent attributes is considered to be its ability to produce extracellular phospholipases that facilitate cellular invasion. Oral candidosis can be treated with polyenes, and azoles, and the more recently introduced echinocandins. However, once administered, the intraoral concentration of these drugs tend to be sub-therapeutic and rather transient due to factors such as the diluent effect of saliva and cleansing effect of the oral musculature. Hence, intra-orally, the pathogenic yeasts may undergo a brief exposure to antifungal drugs. We, therefore, evaluated the phospholipase production of oral C. albicans isolates following brief exposure to sub-therapeutic concentrations of the foregoing antifungals. Materials and methods Fifty C. albicans oral isolates obtained from smokers, diabetics, asthmatics using steroid inhalers, partial denture wearers and healthy individuals were exposed to sub-therapeutic concentrations of nystatin, amphotericin B, caspofungin, ketoconazole and fluconazole for one hour. Thereafter the drugs were removed and the phospholipase production was determined by a plate assay using an egg yolk-agar medium. Results The phospholipase production of these isolates was significantly suppressed with a percentage reduction of 10.65, 12.14, 11.45 and 6.40% following exposure to nystatin, amphotericin B, caspofungin and ketoconazole, respectively. This suppression was not significant following exposure to fluconazole. Conclusions Despite the sub-therapeutic, intra oral, bioavailability of polyenes, echinocandins and ketoconazole, they are likely to produce a persistent antifungal effect by suppressing phospholipase production, which is a key virulent attribute of this common pathogenic yeast.

Frecuencia de dermatomicosis y factores asociados en población vulnerable. Manizales, Colombia / Dermatomicosis frequency and associated factors in vulnerable population. Manizales, Colombia

RESUMEN Objetivo Determinar la frecuencia de las dermatomicosis en personas de diferentes instituciones de atención social en la ciudad de Manizales durante el año 2011. Método Mediante la toma de muestras de los sitios que presentaban algún tipo de lesión sospechosa de ser una micosis cutánea, se hizo un análisis directo con KOH y cultivo en medios de Saboureaud y Mycosel. Se aplicó un instrumento de recolección de información para establecer factores asociados con la presencia de estos microorganismos. Resultados Los hongos levaduriformes encontrados con mayor frecuencia fueron: Candida albicans, Trichosporon sp, y los mohos saprofitos Penicillium sp, fusarium sp; seguido de hongos dermatofitos como: Trichophyton mentagrophytes, Trichophyton rubrum, Epidermophyton floccosum y Microsporum gypseum. Las lesiones secas y descamativas se encontraron con mayor frecuencia. El compartir baños y vivir en hacinamiento y el uso de elementos comunes fueron los factores asociados más importantes en este estudio. Conclusiones Las dermatomicosis son frecuentes en poblaciones vulnerables y se asocian a diferentes factores muy similares a los encontrados en otros estudios de igual naturaleza.(AU)

ABSTRACT Objective To determine the frequency of ringworm in people of different social care institutions in the city of Manizales in 2011. Method Using the sampling sites that had some kind of suspicious lesion from a cutaneous mycosis, direct analysis with KOH and culture media was Saboureaud and Mycosel. An instrument of data collection was used to establish factors associated with the presence of these microorganisms. Results The yeast found most frequently were: Candida albicans, Trichosporon sp and Penicillium molds saprophytes sp, Fusarium sp, followed by dermatophyte fungi such as Trichophyton mentagrophytes, Trichophyton rubrum, Epidermophyton floccosum and Microsporum gypseum. Dry scaly lesions were found more frequently. The shared bathrooms and living in overcrowded and the use of common elements were the most important in this study associated factors. Conclusions Dermatomycoses are common in vulnerable populations and are associated with different very similar to those found in other studies of the same nature factors.(AU)

Candida arthritis in a patient diagnosed with spondyloarthritis

Abstract Candida arthritis is an unusual manifestation that usually affects the knees. A 35-year-old man presented with a 2-month history of pain and swelling in the right knee. Swelling persisted after anti-inflammatory treatment. Peripheric spondyloarthritis was considered, but methotrexate, sulfasalazine, and methylprednisolone did not reduce the swelling. Direct examination of synovial fluid and a culture were positive for Candida albicans. Intravenous and intra-articular amphotericin-B were administered. The arthritis regressed and a culture and direct staining showed negative results. Candida arthritis should be considered in patients with arthritis that is resistant to treatment and prolonged, even if risk factors are absent.