Prevalence of Candida spp. in cervical-vaginal samples and the in vitro susceptibility of isolates

Abstract Vulvovaginal candidiasis (VVC) is an infection of the genital mucosa caused by different species of the genus Candida. Considering the lack of data on this topic in the south of Brazil, this study aimed to assess the prevalence of Candida spp. in the cervical-vaginal mucosa of patients treated at a university hospital in southern Rio Grande do Sul, as well as the etiology and the susceptibility of the isolates against fluconazole, itraconazole, miconazole and nystatin. Samples were collected at the gynecology clinic of the Federal Hospital of the University of Rio Grande, and the isolates were identified using phenotypic and biochemical tests. The susceptibility analysis was performed according to the CLSI M27-A2 protocol. Of the 263 patients included, Candida spp. was isolated in 27%, corresponding to a prevalence of approximately 15% for both VVC and colonization. More than 60% of the isolates were identified as Candida albicans; C. non-albicans was isolated at a rate of 8.6% in symptomatic patients and 14.3% in asymptomatic patients. The prevalence of resistance against fluconazole and itraconazole was 42% and 48%, respectively; the minimal inhibitory concentration of miconazole ranged from 0.031 to 8 µg/mL, and that of nystatin ranged from 2 to >16 µg/mL. The high rate of resistance to triazoles observed in our study suggests the necessity of the association of laboratory exams to clinical diagnosis to minimize the practice of empirical treatments that can contribute to the development of resistance in the isolates.

Impacto de la clindamicina in vitro en la combinación clindamicina - ketoconazol sobre el exopolímero de biopelículas de candida spp de origen urogenital / Impact of in vitro clIndamycIn on the combInatIon of clIndamycIn and ketoconazole on exopolymer of candida spp bIofIlms of urogenItal orIgIn

La mucosa vaginal ha sido utilizada largamente para la administración de antimicrobianos destinados al tratamiento de infecciones endógenas del tracto genital inferior (IETGI) en mujeres embarazadas y no embarazadas. Candida spp elabora biopelículas (BP) y su formación es un proceso complejo que requiere que las células fúngicas establezcan múltiples interacciones con el medio. Las BP están rodeadas por un exopolímero (EPM) que puede restringir la actividad de anticuerpos, la difusión de sustancias y unirse a los antimicrobianos (AM), limitando su acción. Los antimicrobianos (AM) en general y los antimicóticos en particular (AMC) pueden tener dificultades para llegar a las células dentro del EPS. Muchas de las fórmulas que se emplean para el tratamiento empírico usan combinaciones inapropiadas con limitada o nula actividad sobre las biopelículas (BP). La presencia de moléculas que provoquen su inhibición anulando los inductores del EPM o por otro mecanismo, permitirá la actividad del AM específico. Objetivo: demostrar que la actividad de la clindamicina (CLI) en fórmula dual con ketoconazol (KET) actúa sobre BP Candida albicans (CA) y especies no albicans de Candida . (NAC). Métodos: estudiamos la actividad de clindamicina-ketoconazol (CK) sobre la adherencia y dispersión de BP de 8 aislamientos vaginales de CA y 7 de CNA. Se inocularon en 3 tubos con caldo Sabouraud y un dispositivo de vidrio para la formación de la BP según técnica ya descrita. Adherencia: Se incubaron durante 6 horas y se agregó una combinación de CK proveniente del material de óvulos, diluido convenientemente (62,5/260,4 ug/ml), a uno de los tubos de cada aislamiento tomándose como hora 0. Dispersión: esa misma dilución se agregó a otro tubo a las 16 horas. El tercer tubo quedó como testigo sin antimicrobianos. La lectura se efectuó con microscopio óptico a las 24 horas de agregada la combinación CK previa tinción con cristal violeta y se evaluaron con programas fotográficos. Por separado analizamos la actividad de CLI (62,5 ug/ml) y KET (260,4 ug/ml) con técnica similar. Seleccionamos las muestras de 7 pacientes que demostraron candidiasis vulvovaginal (CVV) y las estudiamos con la técnica de capas celulares. Se empleó la combinacion CK para el estudio de la adherencia y dispersión. Resultados: Adherencia se demostró poca influencia de CK en la adherencia con respecto a cada testigo. Dispersión: la influencia de CK se demostró en la mayoría de los aislamientos particularmente en los de CNA que mostraron una mayor presencia de EPM. Las hifas solo se observaron en 1/15 de los aislamientos de Candida spp cuando se agregó CK a las 16 horas. En las BP de las muestras clínicas no aparecieron hifas ni otro elemento micótico en 5/7 con respecto a los testigos. Conclusión: Según estos resultados el uso de una combinación de CK en BP de Candida spp, resulta en una adecuada penetración del AMC demostrada por la dispersión de la BP al cabo de 24 horas. Clindamicina no interfiere con la acción del ketoconazol sino que promovería su actividad anti-candida modificando posiblemente estructuras de superficie y la del EP por inhibición de las moléculas que facilitan la expresión del mismo. In vivo promueve la actividad inmunomoduladora que no se puede demostrar con este modelo in vitro. Su uso combinado en fórmulas duales facilitaría la actividad del AMC sobre Candida spp actuando como inhibidora o modificadora de las BP mediante la dispersión del EPM

The vaginal mucosa has been widely used for administering antimicrobial agents to treat endogenous infections of the lower genital tract in pregnant and non-pregnant women. Candida spp. elaborates biofilms, and its formation is a complex process requiring that fungal cells establish multiple interactions with the medium. Biofilms are surrounded by an exopolymer matrix that can restrict the activity of antibodies, the diffusion of substances, and be associated with antimicrobials, therefore limiting its actions. General antimicrobials and particular anti-mycotic agents can face difficulties to access the cells within the exopolymer matrix. Many formulas used for empirical treatment have improper combinations with limited or null activity on the biofilms. The presence of molecules that cause its inhibition, thus eliminating the exopolymer matrix inducers, or by other mechanism, will allow the specific antimicrobial activity. Objective: To show that the activity of clindamycin used in dual formula with ketoconazole works on Candida albicans biofilm and on non- albicans species of Candida . Methods: We studied the activity of clindamycin and ketoconazole regarding the adherence and dispersion of biofilms from eight vaginal isolates of C. albicans and 7 of non- albicans Candida . The isolates were inoculated in three tubes with Sabouraud agar and a glass device to form the biofilm according to a known technique. Adherence : Each isolate was incubated for a six-hour period and a combination of clindamycin and ketoconazole from the material of ovules was added and conveniently diluted to one of the tubes of each isolate (62.5/260.4 ug/mL), considering 0 hour. Dispersion: The same dilution was added to another tube after 16 hours. The third tube was used as a control without antimicrobials. The reading was carried out with an optical microscope after 24 hours that the clindamycin and ketoconazole combination had been added and colored with crystal violet. They were then evaluated using photographic programs. The activity of clindamycin (62.5 ug/mL) and ketoconazole (260.4 ug/mL) was analyzed alone with a similar technique. We chose vaginal samples from seven patients with vulvovaginal candidiasis and studied them through the cell layer technique. The clindamycin and ketoconazole combination was used for studying the adherence and dispersion. Results: Adherence: Little influence of clindamycin and ketoconazole was seen in adherence regarding each control. Dispersion: The clindamycin and ketoconazole influence was seen in most of the isolates, especially in those of non- albicans Candida that showed higher presence of exopolymer matrix . The hyphae were only seen in 1 of 15 isolates of Candida spp after the clindamycin and ketoconazole were added at the 16th hour. In biofilms of clinical samples, neither hyphae nor mycotic elements were seen in 5 of 7 compared with the controls. Conclusion: According to these results, the use of a clindamycin and ketoconazole combination in biofilms of Candida spp results in proper penetration of the antimicrobial agent, which is seen by the biofilm dispersion during 24 hours. Clindamycin does not interfere with the action of ketoconazole, but it would promote its anti- Candida activity and would possibly modify surface and EP structures through inhibition of the molecules that facilitate its expression. The in vivo model promotes the immunomodulatory activity that in vitro models do not. Its combined use in dual formulas would facilitate the antimicrobial activity on Candida spp, therefore working as an inhibitor or modifier of the biofilms after dispersion of the exopolymer matrix

Susceptibility and molecular characterization of Candida species from patients with vulvovaginitis

Abstract Vulvovaginal candidiasis affects women of reproductive age, which represents approximately 15–25% of vaginitis cases. The present study aimed to isolate and characterize yeast from the patients irrespective of the presentation of clinical symptoms. The isolates were subjected to in vitro susceptibility profile and characterization by molecular markers, which intended to assess the distribution of species. A total of 40 isolates were obtained and identified through the CHROMagar, API20aux and by ITS and D1/D2 regions sequencing of DNAr gene. Candida albicans strains were genotyped by the ABC system and the isolates were divided into two genotypic groups. The identity of the C. albicans, C. glabrata, C. guilliermondii, C. kefyr and Saccharomyces cerevisiae isolates was confirmed by the multilocus analysis. The strains of Candida, isolated from patients with complications, were found to be resistant to nystatin but sensitive to fluconazole, amphotericin B and ketoconazole, as observed by in vitro sensitivity profile. The isolates from asymptomatic patients, i.e., the colonized group, showed a dose-dependent sensitivity to the anti-fungal agents, fluconazole and amphotericin B. However, the isolates of C. albicans that belong to distinct genotypic groups showed the same in vitro susceptibility profile.

O papel dos parceiros sexuais sintomáticos e assintomáticos nas vulvovaginites recorrentes / The role of the symptomatic and asymptomatic sexual partners in the recurrent vulvovaginitis

OBJETIVO: Identificar as espécies predominantes e o papel dos parceiros sexuais na manutenção das vulvovaginites recorrentes por Candida spp. MÉTODOS: Entre agosto de 2007 e março de 2012, foi efetuado estudo prospectivo de 830 pacientes com idades variáveis entre 18 e 65 anos e vaginites fúngicas. Foram excluídos pacientes com diabetes mellitus, síndrome da imunodeficiência adquirida (AIDS), em uso de corticoterapia, antibioticoterapia, hormonoterapia, imunossupressão, duchas vaginais, dispositivos intrauterinos (DIUs) ou espermicidas. A identificação das espécies de Candida foi feita por métodos fenotípicos e genotípicos. O teste do χ2 foi usado para correlacionar a Candida spp. nos parceiros masculinos e a recorrência nas vaginites. RESULTADOS: O agente fúngico foi isolado em um total de 40 mulheres, sendo 24 com vaginites recorrentes, e em 15 dos seus parceiros sexuais, dos quais 10 eram assintomáticos, e 5, sintomáticos. Houve concordância das espécies encontradas no casal em 100% das recorrências. C. albicans (62,4 e 60%), C. glabrata (29,1 e 33,3%) e C. guilliermondii foram as espécies identificadas. Candida tropicalis (4,1%) foi isolada de apenas uma paciente. Nas 16 mulheres restantes que tinham vaginites não complicadas, C. albicans foi isolada em todas. C. glabrata foi isolada em apenas dois de seus parceiros assintomáticos. CONCLUSÃO: Houve predominância de C. albicans; parceiros sintomáticos ou assintomáticos podem ter papel importante como reservatório e fonte de transmissão de leveduras, principalmente nos quadros de vulvovaginites recorrentes. .

PURPOSE: To identify the predominant species and the role of sexual partners in the maintenance of recurrent vulvovaginitis by Candida spp. METHODS: A prospective study of 830 patients aged 18 to 65 years with yeast vaginitis was performed between August 2007 and March 2012. Patients with diabetes mellitus, AIDS or taking corticosteroids, antibiotics or hormone therapy and immunosuppressed patients, patients using vaginal douches, spermicides or intrauterine devices were excluded from the study. Candida species were identified by phenotypic and genotypic methods. The chi-square test was used to correlate the presence of Candida spp. in male partners with the recurrence of vaginitis. RESULTS: The fungal agent was isolated from a total of 40 women, 24 with recurrent vaginitis and from 15 of their sexual partners, 10 of whom were asymptomatic while 5 were symptomatic. There was agreement of the species found in the couple in 100% of recurrences. C. albicans (62.4 and 60%), C. glabrata (29.1 and 33.3%) and C. guilliermondii species were identified. Candida tropicalis (4.1%) was isolated from only one patient. Candida albicans was isolated from the remaining 16 women who had uncomplicated vaginitis. C. glabrata was isolated from only two of the asymptomatic partners. CONCLUSION: There was a predominance of C. albicans and symptomatic or asymptomatic partners can play an important role as a reservoir and source of transmission of yeast, especially in cases of recurrent vulvovaginitis. .

Infecciones vaginales en un Centro de Salud Familiar de la Región Metropolitana, Chile / Vaginal infections in a Family Health Clinic in the Metropolitan Region, Chile

Background: Vaginal infections are a frequent cause for consultation, but their prevalence and etiology vary in different populations. Objectives: To determine the prevalence and etiologies of vaginal infection in women attending a family health center in the Metropolitan Region of Chile. Methods: The microbiological diagnosis was made by wet mount and Gram stain. Diagnosis of trichomoniasis was performed by wet mount, culture and polymerase chain reaction. Results: 101 women aged 15-54, not selected by signs or symptoms of vaginal infection, 46 of them pregnant were included. In 47 women (46.5%), vaginal infections were diagnosed. An association was observed between age and frequency of vaginal infection. The proportion of infections among pregnant and non-pregnant women was similar. The most frequent infections were bacterial vaginosis (16.8%), vulvovaginal candidiasis (11.9%) and co-infections (6.9%). We found 5.9% of intermediate microbiota cases, 3% of trichomoniasis and 2% of aerobic vaginitis. Symptoms of vaginal infection had poor agreement with microbiological findings. Otherwise physical signs had good agreement with the presence of infection, but low to moderate concordance with a specific etiology. Conclusions: We found a high prevalence of vaginal infections in the study population. It is necessary to improve the definitions and criteria of microbiological diagnosis of co-infections and intermediate microbiota, for them to be diagnosed in the clinical practice. More descriptive questionnaires are recommended to enhance the usefulness of clinical examination.

Introducción: Las infección vaginales constituyen un motivo frecuente de consulta, pero su prevalencia y etiología varían en distintas poblaciones. Objetivos: Determinar la prevalencia y tipos de infección vaginal en mujeres atendidas en un centro de salud familiar de la Región Metropolitana. Métodos: El diagnóstico microbiológico fue efectuado mediante examen microscópico al fresco y tinción de Gram y para tricomoniasis examen al fresco, cultivo y reacción de la polimerasa en cadena. Resultados: Se incluyeron 101 mujeres de 15-54 años, no seleccionadas por signos ó síntomas, 46 de ellas embarazadas. En 47 mujeres (46,5%) se diagnosticaron infecciones vaginales. Se observó asociación entre edad y frecuencia de infección vaginal. La proporción de infecciones entre gestantes y no gestantes fue similar. Las infecciones más frecuentes fueron vaginosis bacteriana (16,8%), candidiasis vulvo-vaginal (11,9%) y co-infecciones (6,9%). Se observó 5,9% casos de microbiota intermedia, 3% de tricomoniasis y 2% de vaginitis aeróbica. Los síntomas de infección vaginal tuvieron mala concordancia con los hallazgos microbiológicos. A su vez, los signos físicos tuvieron buena concordancia con la existencia de infección, pero escasa a moderada concordancia con una etiología específica. Conclusiones: Se encontró alta frecuencia de infecciones vaginales. Es necesario mejorar las definiciones y criterios de diagnóstico microbiológico de las co-infecciones y microbiota intermedia, para ser diagnosticadas en la práctica clínica. Se recomienda emplear cuestionarios más descriptivos para mejorar la utilidad del examen clínico.

Evaluación de la disfunción vaginal en mujeres embarazadas sintomáticas y asintomáticas mediante la utilización de los estados vaginales básicos (EVB) y su comparación con el estudio microbiológico convencional / Evaluation of vaginal dysfunction in symptomatic and asymptomatic pregnant women by using the analysis of basic vaginal states (BVS) and its comparison with the conventional microbiological study

Durante el embarazo se producen frecuentemente infecciones del tracto genital inferior asociadas a complicaciones maternas y perinatológicas. El objetivo del trabajo fue evaluar la disfunción vaginal mediante el análisis de los estados vaginales básicos (EVB) por la metodología del balance del contenido vaginal (BACOVA) y compararlo con el estudio microbiológico convencional en el diagnóstico de candidiasis, tricomonosis y vaginosis bacteriana (VB). Entre 2010 y 2012 se estudiaron 1238 pacientes embarazadas; 1046 eran asintomáticas (A) y 192 sintomáticas (S). La prevalencia del EVB I fue del 59,5 % y 26 %, respectivamente. El EVB II se observó en 19,7 % de las mujeres A y en 17,2 % de las S. El EVB III se detectó solamente en las A, en 0,4 %. El EVB IV se observó en 14,4 % de las A y en 38 % de las S. El EVB V se detectó en 6 % de las A y en 18,8 % de las S. En las mujeres A, las levaduras se asociaron a los EVB I y II en el 55,5 % y 23,2 % de los casos, respectivamente; entre las S, alcanzaron el 32,4 % y 31 % de los casos, en igual orden. Las tricomonas se asociaron al EVB I en el 50 % de las A, al EVB IV en el 44,4 % de las S y al EVB V en el 33,3 % de las S. La sensibilidad del BACOVA para detectar levaduras fue 80,4 % en las A y 85,5 % en las S; para detectar tricomonas, del 40 % y 75 %, y para detectar VB, del 100 % en los dos grupos. La especificidad del BACOVA fue 100 % para todos los patógenos en las A y en las S. El estudio de los EVB resultó útil para orientar el diagnóstico a la disfunción vaginal, independientemente de la sintomatología, por lo que se sugiere este estudio como parte del control prenatal. Durante el embarazo se producen frecuentemente infecciones del tracto genital inferior asociadas a complicaciones maternas y perinatológicas. El objetivo del trabajo fue evaluar la disfunción vaginal mediante el análisis de los estados vaginales básicos (EVB) por la metodología del balance del contenido vaginal (BACOVA) y compararlo con el estudio microbiológico convencional en el diagnóstico de candidiasis, tricomonosis y vaginosis bacteriana (VB). Entre 2010 y 2012 se estudiaron 1238 pacientes embarazadas; 1046 eran asintomáticas (A) y 192 sintomáticas (S). La prevalencia del EVB I fue del 59,5 % y 26 %, respectivamente. El EVB II se observó en 19,7 % de las mujeres A y en 17,2 % de las S. El EVB III se detectó solamente en las A, en 0,4 %. El EVB IV se observó en 14,4 % de las A y en 38 % de las S. El EVB V se detectó en 6 % de las A y en 18,8 % de las S. En las mujeres A, las levaduras se asociaron a los EVB I y II en el 55,5 % y 23,2 % de los casos, respectivamente; entre las S, alcanzaron el 32,4 % y 31 % de los casos, en igual orden. Las tricomonas se asociaron al EVB I en el 50 % de las A, al EVB IV en el 44,4 % de las S y al EVB V en el 33,3 % de las S. La sensibilidad del BACOVA para detectar levaduras fue 80,4 % en las A y 85,5 % en las S; para detectar tricomonas, del 40 % y 75 %, y para detectar VB, del 100 % en los dos grupos. La especificidad del BACOVA fue 100 % para todos los patógenos en las A y en las S. El estudio de los EVB resultó útil para orientar el diagnóstico a la disfunción vaginal, independientemente de la sintomatología, por lo que se sugiere este estudio como parte del control prenatal.

Infections of the lower genital tract associated to maternal and perinatal complications frequently occur during pregnancy. The aim of this study was to evaluate vaginal dysfunction through the analysis of basic vaginal states (BVS) using the methodology of balance of the vaginal content (BAVACO) and to compare it with the microbiological study of candidiasis, trichomoniasis and bacterial vaginosis (BV). Pregnant patients (1238) were examined from 2010 to 2012. In asymptomatic (A) (n: 1046) and symptomatic pregnant women (S) (n: 192) BVS I was 59.5% and 26% of the patients, respectively. BVS II was observed in 19.7% of A and in 17.2% of S. BVS III was only detected in A in 0.4%. BVS IV was observed in 14.4% of A and in 38% of S. BVS V was detected in 6% of A and in 18.8% of S. Yeasts were associated to BVS I and II in 55.5% and 23.2% of A, respectively; and in 32.4% and 31% of S, respectively. Trichomonas were associated to BVS I in 50% of A, to IV in 44.4% of S and to V in 33.3% of S. BAVACO susceptibility to detect yeasts was 80.4% and 85.5% in A and S, respectively; 40% and 75% in A and S, respectively, to detect trichomonas and 100% in A and S to detect BV. BAVACO specificity was 100% for all pathogens in A and S. The study of BVS proved useful as a guide to evaluate vaginal dysfunction, regardless of symptomatology. Therefore, this study is recommended as prenatal control. Infections of the lower genital tract associated to maternal and perinatal complications frequently occur during pregnancy. The aim of this study was to evaluate vaginal dysfunction through the analysis of basic vaginal states (BVS) using the methodology of balance of the vaginal content (BAVACO) and to compare it with the microbiological study of candidiasis, trichomoniasis and bacterial vaginosis (BV). Pregnant patients (1238) were examined from 2010 to 2012. In asymptomatic (A) (n: 1046) and symptomatic pregnant women (S) (n: 192) BVS I was 59.5% and 26% of the patients, respectively. BVS II was observed in 19.7% of A and in 17.2% of S. BVS III was only detected in A in 0.4%. BVS IV was observed in 14.4% of A and in 38% of S. BVS V was detected in 6% of A and in 18.8% of S. Yeasts were associated to BVS I and II in 55.5% and 23.2% of A, respectively; and in 32.4% and 31% of S, respectively. Trichomonas were associated to BVS I in 50% of A, to IV in 44.4% of S and to V in 33.3% of S. BAVACO susceptibility to detect yeasts was 80.4% and 85.5% in A and S, respectively; 40% and 75% in A and S, respectively, to detect trichomonas and 100% in A and S to detect BV. BAVACO specificity was 100% for all pathogens in A and S. The study of BVS proved useful as a guide to evaluate vaginal dysfunction, regardless of symptomatology. Therefore, this study is recommended as prenatal control.

Prevalence of Candida albicans and non-albicans isolates from vaginal secretions: comparative evaluation of colonization, vaginal candidiasis and recurrent vaginal candidiasis in diabetic and non-diabetic women / Prevalência de Candida albicans e não albicans isoladas de secreção vaginal: avaliação comparativa entre colonização, candidíase vaginal e candidíase vaginal recorrente em mulheres diabéticas e não diabéticas

CONTEXT AND OBJECTIVE: Vulvovaginal candidiasis (VVC) is caused by abnormal growth of yeast-like fungi on the female genital tract mucosa. Patients with diabetes mellitus (DM) are more susceptible to fungal infections, including those caused by species of Candida. The present study investigated the frequency of total isolation of vaginal Candida spp., and its different clinical profiles - colonization, VVC and recurrent VVC (RVVC) - in women with DM type 2, compared with non-diabetic women. The cure rate using fluconazole treatment was also evaluated. DESIGN AND SETTING: Cross-sectional study conducted in the public healthcare system of Maringá, Paraná, Brazil. METHODS: The study involved 717 women aged 17-74 years, of whom 48 (6.7%) had DM type 2 (mean age: 53.7 years), regardless of signs and symptoms of VVC. The yeasts were isolated and identified using classical phenotypic methods. RESULTS: In the non-diabetic group (controls), total vaginal yeast isolation occurred in 79 (11.8%) women, and in the diabetic group in 9 (18.8%) (P = 0.000). The diabetic group showed more symptomatic (VVC + RVVC = 66.66%) than colonized (33.33%) women, and showed significantly more colonization, VVC and RVVC than seen among the controls. The mean cure rate using fluconazole was 75.0% in the diabetic group and 86.7% in the control group (P = 0.51). CONCLUSION: We found that DM type 2 in Brazilian women was associated with yeast colonization, VVC and RVVC, and similar isolation rates for C. albicans and non-albicans species. Good cure rates were obtained using fluconazole in both groups. .

CONTEXTO E OBJETIVO: Candidíase vulvovaginal (CVV) é causada pelo crescimento anormal de fungos do tipo leveduras na mucosa do trato genital feminino. Pacientes com diabetes mellitus (DM) são mais susceptíveis a infecções fúngicas, incluindo por espécies de Candida. O presente estudo investigou a frequência de isolamento total de Candida spp. vaginal, e diferentes quadros clínicos (CVV e CVV recorrente- CVVR) em mulheres com DM tipo 2 comparadas às não diabéticas. A razão de cura do tratamento com fluconazol também foi avaliada. TIPO DE ESTUDO E LOCAL: Estudo transversal realizado no sistema público de saúde de Maringá, Paraná, Brazil. MÉTODO: O estudo envolveu 717 mulheres de 17-74 anos de idade e, destas, 48 (6,7%) tinham DM 2 (média de 53,7 anos), independentemente de sinais e sintomas de CVV. As leveduras foram isoladas e identificadas por métodos fenotípicos clássicos. RESULTADOS: No grupo de não diabéticas (controle), leveduras vaginais totais foram isoladas em 79 (11,8%) mulheres, e no grupo de diabéticas, em 9 (18,8%) (P = 0,000). O grupo de diabéticas mostrou mais mulheres sintomáticas (CVV + CVVR = 66,66%) do que colonizadas (33.33%), e significativamente mais colonização, CVV e CVVR, que as controle. A razão média de cura com fluconazol foi de 75.0% no grupo diabéticas e 86.7% no controle (P = 0.51). CONCLUSÃO: Nós encontramos que DM 2 em mulheres brasileiras associou-se com colonização vaginal por leveduras, CVV e CVVR, razão similar de isolamento de C. albicans e espécies não albicans. Boa taxa de cura foi obtida com fluconazol em ambos os grupos.

Associação entre cultura de secreção vaginal, características sociodemográficas e manifestações clínicas de pacientes com diagnóstico de candidíase vulvovaginal / Association between vaginal secretion culture, socio-demographic characteristics and clinical manifestations of patients with vulvovaginal candidiasis

OBJETIVO: Investigar a etiologia, o perfil epidemiológico de pacientes com candidíase vulvovaginal (CVV) e possíveis fatores predisponentes. MÉTODOS: Secreção vaginal das pacientes foi semeada em ágar Sabouraud e amostras de leveduras foram isoladas e identificadas por Polymerase Chain Reaction (PCR). Dados demográficos, clínicos e fatores predisponentes foram obtidos por meio de questionário. Para análise estatística, foram utilizados os testes t de Student, Fischer e do χ², com auxílio do software Statistical Package for the Social Sciences (SPSS), com nível de significância de 5%. RESULTADOS: Foram avaliadas 69 pacientes, com idade entre 15 e 52 anos, predominando mulheres brancas (79,7%), com escolaridade de nível superior completo (58%), casadas (56,5%) e com vida sexual ativa (97,1%). Dentre elas, 34,8% eram gestantes, 7,2% diabéticas, 1,4% soropositivas para AIDS e 36,2% usavam anticoncepcional oral. Antibioticoterapia recente foi citada por 13% das pacientes, uso de antifúngico por 5,8% e de antitricomonas por 1,4%. Uso de corticosteroides foi relatado por 2,9% das participantes e de antineoplásicos, por 1,4%. Fluxo vaginal e prurido foram as principais queixas apresentadas, respectivamente, por 97,1 e 73,9% das pacientes, seguido de ardência (63,8%) e hiperemia (63,8%). Quando presente, o fluxo foi majoritariamente branco (88,1%) ou grumoso (86,6%). O diagnóstico foi confirmado pela cultura em 55 (79,7%) pacientes, sendo 4 casos de infecção mista. A espécie prevalente foi C. albicans, seguida por um caso de C. glabrata, que foi encontrada em mais duas pacientes em associação com C. albicans. Nas outras duas infecções polimicrobianas, C. lusitaniae foi isolada com C. albicans. CONCLUSÕES: Embora a positividade da cultura tenha sido alta e os dados clínicos de CVV sejam característicos, a sintomatologia não é patognomônica. C. albicans é a espécie prevalente, mas deve-se atentar para a ocorrência de outras espécies na etiologia de CVV, como a emergência de C. lusitaniae.

PURPOSE: To investigate the etiology and the epidemiological profile of patients with vulvovaginal candidiasis (VVC) and predisposing factors. METHODS: Vaginal secretions were streaked in Sabouraud agar and yeast samples were isolated and identified by Polymerase Chain Reaction (PCR). Demographic and clinical data were obtained with a questionnaire. For statistical analysis, the Student's t-test, the χ² and Fischer tests were applied as needed using the Statistical Package for Social Sciences (SPSS) software, with the level of significance set at 5%. RESULTS: Sixty-nine patients aged from 15 to 52 years were evaluated. They were predominantly white (79.7%), with higher education (58%), married (56.5%) and sexually active (97.1%). Among them, 34.8% were pregnant, 7.2% diabetic, 1.4% seropositive for AIDS, and 36.2% were using oral contraceptives. Recent antibiotic therapy was mentioned by 13% of the patients, and antifungal or anti-trichomonas therapy was mentioned by 5.8 and 1.4% of the patients, respectively. Corticosteroid use was reported by 2.9% and antineoplastic by 1.4%. Vaginal discharge and itching were the main complaints (97.1 and 73.9%), followed by burning (63.8%) and erythema (63.8%). When present, the vaginal flow was predominantly white (88.1%) or lumpy (86.6%). The diagnosis was confirmed by culture in 55 (79.7%) patients, with mixed infections in 4 patients. The most prevalent species was C. albicans, followed by C. glabrata (one monoinfection and two mixed infections with C. albicans). C. lusitaniae and C. albicans were also identified in mixed infections (two patients). CONCLUSION: Despite the high culture positivity and clinical data characteristic of VVC, the symptoms were not pathognomonic. C. albicans is the most prevalent species, but other species are also involved in VVC etiology, such as the emergence of C. lusitaniae.

Teste de sensibilidade ao quefir de cepas de Candida sp. isoladas de vulvovaginites / Test of sensitivity to kefir of Candida spp. strainsisolated from vulvovaginitis

A candidíase vulvovaginal é uma inflamação da mucosa genital que se desenvolve em decorrência de infecção por leveduras do gênero Candida, as quais são habitantes normais do canal vaginal. O quefir é um probiótico que se apresenta em grãos ou grumos, formado por diversos microrganismos, como por exemplo, várias espécies de Lactobacillus e leveduras, que vivem em um complexo sistema de simbiose adsorvidos em uma matriz de polissacarídeos secretados pelo mesmo. O presente estudo foi realizado com o objetivo de isolar e identificar leveduras de vulvovaginites recorrentes e testar in vitro a ação do quefir sobre estes microrganismos. Para o isolamento e identificação utilizou-se CHROMagar-Candida e confirmação bioquímica. O antibiograma foi realizado pela técnica de poços. Das 44 amostras coletadas, 40,91% foram positivas para Candida sp., as quais mostraram prevalência das espécies não-albicans (52,63%) sobre as espécies de C. albicans. No antibiograma, houve uma sensibilidade de 68,42% de ambas as populações à suspensão de quefir, enquanto para o mesmo, veiculado em base não-iônica, houve redução do espectro de ação. Concluiu-se que a suspensão foi mais eficaz em relação à sua associação com base não iônica, o que torna inviável a veiculação desta formulação.

Vulvovaginal candidiasis is an inflammation of the genital mucosa that develops as a result of infection by Candida yeast species, which are normal inhabitants of the vagina. Kefir is a fermented-milk probiotic produced in the form of grains or flakes, by a symbiotic complex of microorganisms, among them several species of lactic-acid bacteria and yeasts, which live encapsulated in a matrix of polysaccharides secreted by the former. This study was conducted to isolate and identify yeasts from recurrent vulvovaginitis and test in vitro the action of kefir on these microorganisms. For the isolation and identification of Candida organisms, CHROMagar-Candida and biochemical confirmation were employed. Overall antimicrobial susceptibility was determined by the agar well diffusion technique. Of the 44 samples, 40.91% were positive for Candida spp., which showed a prevalence of non-albicans species (52.63%) over the species C. albicans. In the antibiogram (in vitro susceptibility test), there was a sensitivity of 68.42% of both populations to the suspension of kefir alone, while for kefir carried in a non-ionic cream base, the spectrum of action was reduced. It was concluded that the suspension was more effective alone than when combined with a non-ionic base, making it impractical to use this formulation.

Posible papel de Bacteroides fragilis enterotoxigénico en la etiología de la vaginitis infecciosa / Possible role of enterotoxigenic Bacteroides fragilis in the etiology of infectious vaginitis

La vaginitis es un trastorno ginecológico frecuente producido por distintas causas, algunas de las cuales permanecen desconocidas. Bacteroides fragilis es el anaerobio más importante en bacteriología clínica. Algunas cepas son enterotoxigénicas y se asocian con síndromes intestinales y extraintestinales. Recientemente han sido aisladas de pacientes con vaginitis. En este trabajo se planteó investigar la posible asociación de B. fragilis enterotoxigénico con la vaginitis infecciosa. Fueron procesadas 265 muestras de exudado vaginal. 202 de mujeres sintomáticas y 63 mujeres sanas. La identificación de los microorganismos se realizó por métodos convencionales. En 31,2% de las pacientes sintomáticas se identificaron: Gardnerella vaginalis, Candida albicans, Mobiluncus, Mycoplasma hominis, Ureaplasma urealyticum y Streptococcus agalactiae. En 27 pacientes sintomáticas y en 5 mujeres sanas se identificó B. fragilis. Estas cepas fueron cultivadas en medio líquido e incubadas durante 48 h a 36° C en anaerobiosis. La toxicidad en los sobrenadantes se ensayó en células HT-29. 18 cepas de B. fragilis aisladas de pacientes sintomáticas fueron enterotoxigénicas, ya que indujeron alteraciones en la monocapa celular y en las células. No se identificó en mujeres sanas (P<0,05). 77,7% de las cepas de B. fragilis enterotoxigénicas no se encontraron asociadas con otros patógenos específicos. Este hecho sugiere que pudiera ser un agente causante de vaginitis, ya que el efecto de la enterotoxina sobre la E-cadherina del epitelio vaginal podría facilitar la invasión y su posible papel patógeno en la vagina. Esta es la primera investigación que asocia a Bacteroides fragilis enterotoxigénico como posible causa de vaginitis infecciosa.

Vaginitis is a common gynecologic disorder. It is due to several causes, some even unknown. Bacteroides fragilis is the most important anaerobe in clinical bacteriology, some strains of this group are notable for being enterotoxigenic and they have been associated with intestinal and extraintestinal syndromes. They have recently been isolated from patients with vaginitis. The purpose of this study was to investigate a possible association of enterotoxigenic B. fragilis with infectious vaginitis. 265 samples of vaginal exudate were processed, 202 from symptomatic patients and 63 healthy women. The identification of the microorganisms was carried out by conventional methods. In 31.2% of symptomatic patients were identified: Gardnerella vaginalis, Mobiluncus, Candida albicans, Mycoplasma hominis, Ureaplasma urealyticum and Streptococcus agalactiae. B. fragilis was identified in 27 symptomatic patients and 5 healthy women. These strains were cultivated in liquid medium and incubated during 48 h at 36°C in anaerobe chambers. Supernatant activity was assayed in HT-29 cells. Eighteen B. fragilis strains isolated from symptomatic patients were enterotoxigenic, because induced alterations in target cell morphology. It was not identified in healthy women (P<0.05). 77.7% of enterotoxigenic B. fragilis strains were not associated with other specific pathogens. This fact suggests that enterotoxigenic B. fragilis could be a cause for vaginitis. The effect of enterotoxin on E-cadherin of vaginal epithelium could facilitate invasion and its possible pathogenic role in the vagina. This is the first report that associates enterotoxigenic Bacteroides fragilis as a possible cause of infectious vaginitis.

Importancia del estudio del balance del contenido vaginal (BACOVA) en el control preventivo de las trabajadoras sexuales / Importance of studying the balance of vaginal content (BAVACO) in the preventive control of sex workers

El objetivo de este trabajo fue estudiar a un grupo de 229 trabajadoras sexuales de Comodoro Rivadavia (Chubut), atendidas en centros públicos de salud de dicha ciudad, mediante la aplicación del método conocido como balance del contenido vaginal (BACOVA). Este método comprende el estudio morfológico de la microbiota vaginal, como así también de la reacción infamatoria. Incluye el análisis del contenido vaginal en fresco y por tinciones de Gram y de Giemsa, de modo de integrar la exploración de todo el panorama biológico. El 35,37 % de estas mujeres presentó microbiota normal (MN); el 15,72 %, microbiota intermedia (MI); el 23,14 %, vaginosis bacteriana (VB) y el 10,48 %, vaginitis microbiana inespecífca (VMI). Los casos de vaginitis por levaduras y por Trichomonas vaginalis comprendieron el 8,30 % y 6,99 % de las mujeres, respectivamente. Se observó el desplazamiento de la MN hacia una MI, que se correspondió con el predominio de bacterias corineformes. Por otra parte, no se reconoció un marcado desequilibrio del contenido vaginal ante la colonización e infección por levaduras o por T. vaginalis: el 48 % de los casos de estas vaginitis convencionales no presentaron reacción infamatoria vaginal (RIV). El 24,89 % de los casos de MN presentaron una signifcativa RIV, y en más del 50 % de las mujeres se diagnosticaron disfunciones vaginales en ausencia de sintomatología. Estos resultados se podrían asociar a un incremento del riesgo gineco-obstétrico, lo que afecta la salud sexual y reproductiva de la población estudiada.

The aim of this work was to study the vaginal microenvironment in sex workers from Comodoro Rivadavia, Chubut. For that purpose, BAVACO procedures were applied. A total of 229 female sex workers attended public health centers. Vaginal secretions were analyzed by Gram and Giemsa stains. The following results were obtained: normal microbiota 35.37 %, intermediate microbiota 15.72 %, bacterial vaginosis 23.14 %, microbial nonspecifc vaginitis, Donders'"aerobic vaginitis" 10.48 %, yeast vulvovaginitis 8.30 %, and trichomoniasis 6.99 %. The intermediate microbiota was characterized by a decrease in the number of lactobacilli and the presence of diphtheroid bacilli cell types. The population studied shared increased values of vaginal dysfunctions. These results are considered risk factors for obstetric and gynecologic diseases.

Candida species isolated from the vaginal mucosa of HIV-infected women in Salvador, Bahia, Brazil

BACKGROUND: Vulvovaginal candidiasis (VVC) is the second most common vaginal infection. HIV-infection is a risk factor for this infection. OBJECTIVE: To determine the frequency of VVC and to describe the main Candida species isolated and their susceptibility to antifungal drugs in HIV-infected patients, compared to HIV-uninfected women in Salvador, Brazil. METHODS: Cross-sectional study including a group of 64 HIV-infected women and 76 uninfected women, followed up at the AIDS reference center and at the Gynecological Clinic of Escola Bahiana de Medicina e Saúde Pública (Salvador, Bahia, Brazil). RESULTS: Frequency of Candida spp. was higher in HIV-infected women (29.7 percent) than in HIV-uninfected controls (14.5 percent) (p = 0.02). The odds ratio value for vulvovaginal candidiasis in HIV-infected patients was 2.6 (95 percent CI: 1.07 - 6.32 p = 0.03). Candida albicans was the most commonly isolated species in both HIV-infected (52.3 percent) and uninfected women (85.7 percent), followed by C. parapsolis in 17.6 percent and 14.3 percent, respectively. In HIV-infected women, C. glabrata, C. parapsilosis, and a coinfection of C. albicans and C. glabrata were also identified. There was no significant difference between Candida species isolated from the vaginal mucosa of women with VVC and colonization of the vaginal mucosa of HIV-infected and HIV-uninfected women. One C. glabrata isolate from an HIV-infected patient was resistant to fluconazole and other two isolates exhibited a dose-dependent susceptibility. CONCLUSION: Our results confirm a higher frequency of Candida spp. isolated from the vaginal mucosa of HIV-infected women and a broader spectrum of species involved. Only Candida glabrata isolates showed decreased susceptibility to fluconazole.

Identificação por PCR e sensibilidade a antifúngicos de isolados clínicos vaginais de Candida sp / Identification by PCR and antifungal susceptibility of vaginal clinical Candida sp isolates

INTRODUÇÃO: A candidíase vaginal é uma condição que afeta um grande número de mulheres em idade fértil. Candida albicans é a espécie mais frequentemente isolada de secreção vaginal, entretanto, outras espécies mais resistentes às drogas antifúngicas podem ser isoladas de amostras clínicas vaginais. MÉTODOS: Foram identificadas as espécies de 30 isolados vaginais de Candida sp por PCR utilizando o primer universal ITS4 e primers espécie-específicos para C. albicans, C. glabrata, C. tropicalis e C. krusei. A sensibilidade destes isolados frente à anfotericina B, fluconazol e voriconazol foi determinada pelo método de macrodiluição M27-A2 do CLSI. RESULTADOS: Através dos ensaios de PCR, 28 isolados foram caracterizados como C. albicans e 2 isolados apresentaram amplificação para os primers específicos de C. albicans e C. glabrata. A concentração inibitória mínima para anfotericina B variou de 0,03µg/mL a 0,25µg/mL, para o fluconazol de 0,125µg/mL a 16µg/mL e para o voriconazol de 0,03µg/mL a 0,25µg/mL. CONCLUSÕES: A identificação de Candida ao nível de espécie através de ensaios de PCR deve ser relevante para o gerenciamento clínico destas infecções.

INTRODUCTION: Vaginal candidiasis is a condition that affects innumerous fertile women. Candida albicans is the most frequently isolated species from vaginal discharge; however, other different species that are more resistant to antifungal drugs can be identified in vaginal clinical samples. METHODS: The species of 30 vaginal Candida isolates was identified by PCR using the universal ITS4 primer and species-specific primers for C. albicans, C. glabrata, C. tropicalis and C. krusei. The sensitivity pattern to amphotericin B, fluconazole and voriconazole was assessed using the CLSI M27-A2 macrodilution method. RESULTS: The PCR assay revealed 28 C. albicans and 2 samples showed amplification for C. albicans and C. glabrata primers. The minimum inhibitory concentration for amphotericin B ranged from 0.03µg/mL to 0.25µg/mL, for fluconazole from 0.125µg/ml to 16µg/mL and for voriconazole from 0.03µg/mL to 0.25µg/m. CONCLUSIONS: Identification at Candida species level by PCR assay could be relevant for clinical management of these infections.

Candidíase vaginal recorrente: manejo clínico / Recurrent vaginal candidiasis: clinical management

A candidíase vulvovaginal (CVV) é patologia extremamente frequente, atingindo 75% das mulheres em alguma fase da vida. Cerca de 5% delas terão candidíase vulvovaginal recorrente (CVVR), definida como quatro ou mais episódios a cada 12 meses. Apesar de existirem fatores de risco conhecidos, a maioria dos episódios ocorre na sua ausência. Com base na apresentação clínica, microbiologia e fatores do hospedeiro, ela é classificada em complicada e não-complicada. Um espectro de sintomas comuns a outras patologias, e a ausência de métodos rápidos, simples e baratos de diagnóstico, tornam seu manejo um desafio, mesmo para os mais experientes. O exame padrão-ouro para confirmar a infecção é cultura em meio específico. O presente estudo tem por objetivo revisar os métodos diagnósticos da CVV e descrever o manejo da CVVR.

Vulvovaginal candidiasis (VVC) is an extremely frequent pathology, affecting 75% of women in some phase of life. About 5% of them will have recurrent vulvovaginal candidiasis (RVVC), defined as four or more episodes every 12 months. In spite of known risk factors, most of the episodes happen in its absence. Based on the clinical presentation microbiology and factors of the host, it is classified in complicated and non-complicated. A spectrum of symptoms common to other pathologies, and the absence of fast, simple and inexpensive methods of diagnosis, turns it into a challenge even for experts. The gold standard exam to confirm the infection is a culture in specific medium. The present study has the objective of revising the diagnosis methods of VVC and to describe the management of RVVC.

Vaginosis citolítica: un diagnóstico diferencial poco frecuente de vulvovaginitis micótica a repetición / Cytolytic vaginosis: a rare differential diagnosis of fungal recurrent vulvovaginitis

Los síntomas compatibles con vaginitis y vaginosis, son la causa más frecuente de consulta al ginecólogo. La descarga vaginal abundante y el prurito, constituyen la primera sospecha de estas patologías. Sin embargo, debemos tener presente que el aumento significativo de lactobacilos, conocido como lactobacilosis, puede expresarse también con los mismos síntomas y signos de una vulvovaginitis micótica. Si esta condición se mantiene en el tiempo, puede provocar un grado de acidez anormal en la vagina, y puede llevar a una lisis de las células epiteliales que se conoce como vaginosis citolítica. Este diagnóstico, aunque es poco frecuente, debemos sospecharlo especialmente, en aquellas mujeres que relatan tener múltiples consultas por candidiasis vaginal, con escasa o nula respuesta frente a la terapia antimicótica.

The symptoms consistent with vaginitis and vaginosis are the most common cause of the visit to the gynecologist. Heavy vaginal discharge and pruritus are the first suspicion of this pathology. However, the significant increase in lactobacilli, known as lactobacilosis can be expressed also with the same symptoms and signs of a fungal vulvovaginitis. If this condition is maintained overtime, can cause abnormal acidity in the vagina, and may lead to epithelial cell lysis, which is known as cytolytic vaginosis. This diagnosis, although rare, should be suspected especially in women who reported having multiple consultations for vaginal candidiasis, with little or no response to antifungal therapy.

Candida glabrata: un oportunista emergente en vulvovaginitis / Candida glabrata: an emergent opportunist in vulvovaginitis

Introducción: El género Candida comprende varias especies; en años recientes algunas como Candida glabrata ha incrementado su frecuencia con trascendencia clínica. Material y métodos: Se realizó un estudio para determinar la frecuencia de Candida glabrata en 468 pacientes con sintomatología clínica para candidosis vulvovaginal, así como la sensibilidad de la misma a fluconazol por métodos de difusión en agar con sensidiscos y microdilución en caldo. Resultados: La frecuencia para esta especie fue de 12.6 %. La resistencia de Candida glabrata al tratamiento con fluconazol se corroboró en este estudio: 68.2 % de las cepas fue resistente en pruebas de placas (sensidiscos) y 51.2 % en prueba de microdilución en caldo (método NCLSI), con una concentración mínima inhibitoria de 16 μg/ml. Conclusiones: La frecuencia de Candida glabrata se ha incrementado y presenta resistencia a los tratamientos habituales, lo que influye en la persistencia y recurrencia de infecciones genitales y sistémicas.

BACKGROUND: Candida genus has various species. The incidence of C. glabrata has presented itself with more frequency over the past years with clinical importance. METHODS: A case study was made to determine the frequency of C. glabrata in 468 patients who presented clinical symptomatology for vulvovaginal candidiasis and the in vitro response for fluconazole using two methods: diffusion in agar plates and microdilution in liquid medium [NCLSI (NCCLS) method]. RESULTS: The frequency for this specie was 12.6%, almost double the frequency observed 10 years ago. The resistance of C. glabrata to fluconazole treatment was confirmed in this study, representing 68.2% resistance in all strains on test plates and 51.2% on NCLSI method with a MIC of 16 microg/ml. Conclusions: The frequency of Candida glabrata has increased over the past years. It presents resistance to usual treatments, which promotes the persistence and recurrence of genital and systemic infections.

Frequência de leveduras em fluido vaginal de mulheres com e sem suspeita clínica de candidíase vulvovaginal / Frequency of yeasts in vaginal fluid of women with and without clinical suspicion of vulvovaginal candidiasis

OBJETIVO: estudar a candidíase vulvovaginal em mulheres com e sem suspeita clínica a partir de fluido vaginal, identificando frequência de Candida spp. e associando a fatores de risco intrínsecos e extrínsecos. MÉTODOS: foram coletadas 286 amostras de pacientes atendidas em clínicas e postos de saúde entre Agosto de 2005 e Agosto de 2007. Foram 121 mulheres com suspeita e 165 sem suspeita clínica. Com zaragatoas estéreis, as amostras foram coletadas, transportadas ao laboratório em solução fisiológica 0,85 por cento, semeadas em CHROMagar Candida e em meio ágar Sabouraud 4 por cento com cloranfenicol. Foram realizados os procedimentos clássicos para identificação: macro e micromorfologia, zimograma e auxanograma. Os dados obtidos foram analisados através de testes de frequência e tabelas de contingência (χ2). RESULTADOS: Um total de 47,9 por cento das mulheres com suspeita clínica obteve confirmação de candidíase pelos exames laboratoriais. Das pacientes sem suspeita clínica (Grupo Controle), 78,2 por cento foram negativas para candidíase vulvovaginal pelos testes laboratoriais. Candida albicans foi a espécie prevalente com 74,5 por cento dos casos. Foram encontradas diferenças significativas para os casos positivos, de acordo com as pacientes das duas cidades avaliadas (p<0,05). O vestuário foi um aspecto diferencial encontrado entre as duas populações estudadas. CONCLUSÕES: a presença de fatores predisponentes não define, seguramente, a candidíase vulvovaginal. A localização geográfica tem mostrado ser um fator relevante na distribuição dos eventos. O tipo de vestuário pode ser uma das razões. O cultivo de amostras do conteúdo vaginal, seguida de identificação do micro-organismo, é importante.

PURPOSE: to study vulvovaginal candidiasis from the vaginal fluid of women with and without clinical suspicion, identifying the frequency of Candida spp., and associating it with intrinsic and extrinsic risk factors. METHODS: a total of 286 samples from patients attended in private practices and public health units from August 2005 to August 2007 were collected, being 121 women under clinical suspicion and 165, without. The samples were collected with sterile swabs, taken to the laboratory in 0.85 percent physiological solution, and then seeded in CHROMagar Candida and in 4 percent agar Sabourad with chloramphenicol. Classical identification procedures were carried out: macro and micromorphology, zymogram and auxanogram. Data obtained were analyzed by frequency tests and contingency tables (χ2). RESULTS: a total of 47.9 percent of the women under clinical suspicion got confirmation of candidiasis by the laboratorial tests. Among the patients without clinical suspicion (Control Group), 78.2 percent were vulvovaginal candidiasis negative according to the laboratorial tests. Candida albicans was the prevalent strain in 74.5 percent of the cases. There were significant differences among the positive cases, according to the patients from the two cities evaluated (p<0.05). Clothing was one differential aspect found among the two populations studied. CONCLUSIONS: the presence of predisposing factors does not necessarily define vulvovaginal candidiasis. Geographical localization has shown to be a relevant factor in the distribution of events. The type of clothing may be one of the reasons for it. Culture of samples from the vaginal contents, followed by microorganisms' identification, can be important.

Identificación rápida y sensibilidad a toxinas killer de levaduras aisladas de micosis no sistémicas / Rapid identification and susceptibility to killer toxins of yeasts isolated from non-systemic mycoses

La identificación rápida y segura de los agentes etiológicos y el desarrollo de nuevos antifúngicos con blancos de acción más específicos resultarán en tratamientos de las micosis más efectivos y menos lesivos. Mediante un método molecular rápido (ITS1-5.8S ADNr-ITS2 PCR-RFLP) se identificaron 53 aislamientos de levaduras provenientes de infecciones no sistémicas registradas en hospitales públicos de la ciudad de Neuquén y en un centro oftalmológico de Buenos Aires durante el año 2005. Adicionalmente y utilizando el método de inhibición del crecimiento en placa, se evaluó la sensibilidad de estas levaduras a toxinas killer producidas por levaduras indígenas de la Patagonia y por cepas de referencia. Ocho especies de levaduras fueron identificadas entre los aislamientos clínicos: Candida albicans (52%) , Candida parapsilosis (17%) , Candida tropicalis (10%) , Candida krusei (5%) , Candida glabrata (4%) , Candida guilliermondii (4%) , Kluyveromyces lactis (4%) y Saccharomyces cerevisiae (4%) . El 69% de los aislamientos de la especie mayoritaria, C. albicans, se relacionó con infecciones vaginales. Por otra parte, el 61% de las levaduras provenientes de infecciones oculares correspondió a la especie C. parapsilosis. En las condiciones de ensayo, las toxinas producidas por las levaduras killer indígenas DVMais5 y HCMeiss5 pertenecientes a las especies Pichia anomala y P. kluyveri, respectivamente, exhibieron el mayor espectro de acción sobre las levaduras aisladas de materiales clínicos.

The use of quick and reliable yeast identification methods, as well as the development of new antifungal agents with more specific targets, will enable a more efficient treatment of mycoses. In the present work, a total of 53 clinical isolates obtained from non-systemic infections in Neuquén Hospitals and an ophthalmologic clinic in Buenos Aires during 2005, were identified by means of a rapid molecular method (ITS1-5.8S ADNr-ITS2 PCR-RFLP). Additionally, the killer susceptibility of the isolates was tested against reference and indigenous killer yeasts on plate tests. Eight yeast species were identified among the clinical isolates: Candida albicans (52%), Candida parapsilosis (17%), Candida tropicalis (10%), Candida krusei (5%), Candida glabrata (4%) , Candida guilliermondii (4%) , Kluyveromyces lactis (4%) and Saccharomyces cerevisiae (4%) . Sixty-nine percent of the isolates corresponding to the predominant species ( C. albicans) were related to vaginal infections. On the other hand, 61% of the yeasts associated with ocular infections were identified as C. parapsilosis. Two indigenous killer isolates DVMais5 and HCMeiss5, belonging to Pichia anomala and P. kluyveri respectively, exhibited the broadest killer spectrum against clinical isolates.

Antifungal activity of the extracts and saponins from Sapindus saponaria L

Extracts from the dried pericarp of Sapindus saponaria L. (Sapindaceae) fruits were investigated for their antifungal activity against clinical isolates of yeasts Candida albicans and C. non-albicans from vaginal secretions of women with Vulvovaginal Candidiasis. Four clinical isolates of C. albicans, a single clinical isolated of each of the species C. parapsilosis, C. glabrata, C. tropicalis, and the strain of C. albicans ATCC 90028 were used. The hydroalcoholic extract was bioactivity-directed against a clinical isolate of C. parapsilosis, and showed strong activity. The n-BuOH extract and one fraction showed strong activity against all isolates tested. Further column-chromatography on silica gel separation of this fraction afforded two pure triterpene acetylated saponins: 3-O-(4-acetyl-beta-D-xylopyranosyl)-(1->3)-alpha-Lrhamnopyranosyl-(1->2)-alpha-L-arabinopyranosyl-hederagenin (1) and 3-O-(3,4-di-acetyl-beta-D-xylopyranosyl)-(1->3)-alpha-L-rhamnopyranosyl-(1->2)-alpha-L-arabynopyranosyl-hederagenin (2). The structures of the compounds were based on spectral data (¹H and 13C NMR, HSQC, HMBC and MS), and on with literature. The saponins isolated showed strong activity against C. parapsilosis.

Extratos do pericarpo de frutos de Sapindus saponaria L. (Sapindaceae) foram testados para a atividade antifúngica sobre isolados clínicos de leveduras de Candida albicans e C. não-albicans obtidos de secreção vaginal de mulheres com Candidíase Vulvovaginal. Foram avaliados quatro isolados clínicos de C. albicans, um de cada uma das espécies C. glabrata, C. parapsilosis, C. tropicalis e uma cepa referência de C. albicans ATCC 90028. O extrato hidroalcoólico foi biomonitorado contra um isolado clínico de C. parapsilosis, apresentando forte atividade. O extrato butanólico e uma fração apresentaram forte atividade contra todos os isolados testados. Posterior análise desta fração via cromatografia em sílica gel (CHCl3:CH3OH, 1:1, v/v) resultou no isolamento de duas saponinas triterpênicas puras mono e diacetiladas, 3-O-(4-O-acetil-O-beta-D-xilopiranosil)-(1 -> 3)-alfa-L-ramnopiranosil-(1 -> 2)-alfa-L-arabinopiranosil-hederagenina (1) e 3-O-(3,4-di-O-acetil-beta-D-xilopiranosil)-(1 -> 3)-alfa-L-ramnopiranosil-(1 -> 2)-alfa-L-rabinopiranosil-hederagenina (2) respectivamente. A elucidação estrutural das substâncias foi baseada em dados espectrais (RMN de ¹H e de 13C, HSQC, HMBC, ESI/MS) e comparados com dados da literatura. As saponinas triterpênicas isoladas (1) e (2) apresentaram forte atividade contra C. parapsilosis.