Combining Essential Oils with Each Other and with Clotrimazole Prevents the Formation of Candida Biofilms and Eradicates Mature Biofilms.

Fungal infections by Candida spp. are opportunistic and most often occur in individuals with some predisposing factor. Essential oils (EO) have anti-Candida potential, being a therapeutic alternative to be explored, especially for superficial and mucosal candidiasis. The objective was to analyze the synergistic potential between the EO of Citrus limon, Cupressus sempervirens, Litsea cubeba and Melaleuca alternifolia, and each of them with clotrimazole, to inhibit in vitro the formation and eradication of Candida spp. biofilms. Added to this, the survival of Caenorhabditis elegans was evaluated after exposure to EO, clotrimazole and their synergistic combinations. Anti-Candida activity was determined by microdilution for the substances alone and in EO−EO and EO−clotrimazole combinations. The combinations were performed by the checkerboard method, and the reduction in the metabolic activity of biofilms was determined by the viability of MTT/menadione. C. elegans larvae survival was evaluated after 24 h of exposure to EO, clotrimazole and synergistic combinations. The minimum inhibitory concentration (MIC) of EO ranged from 500 to >4000 µg/mL. The lowest MIC (500 µg/mL) was for C. sempervirens and L. cubeba on a C. krusei isolate; for clotrimazole, the MIC ranged from 0.015 to 0.5 µg/mL. Biofilm inhibition and eradication both ranged from 1000 to >4000 µg/mL. The lethal concentration (LC50) of C. limon, L. cubeba and M. alternifolia was 2000 µg/mL for C. elegans, while for C. sempervirens and clotrimazole, it was not determined within the concentration limits tested. In combination, more than 85% of the larvae survived M. alternifolia−clotrimazole, M. alternifolia−L. cubeba, C. sempervirens−clotrimazole and C. sempervirens−C. limon combinations. This study is the first, to our knowledge, to present a synergistic relationship of EO−EO and EO−clotrimazole combinations on Candida spp. biofilms.

Inanimate Surfaces and Air Contamination with Multidrug Resistant Species of Staphylococcus in the Neonatal Intensive Care Unit Environment.

BACKGROUND: Contamination of the hospital environment with multi-resistant (MDR) Staphylococcus increases the risk of infection. The aim of this study is to identify the MDR species of Staphylococcus on inanimate surfaces, in air, and in clinical samples, and analyze the risk factors that correlate with the occurrence of infections in a Neonatal Intensive Care Unit. METHODS: Samples of inanimate surfaces and air were taken using a premoistened swab (0.9% sodium chloride) and spontaneous air sedimentation, respectively. The clinical isolates were recovered from infected neonates. The isolates (environmental and clinical) were identified by matrix-assisted laser desorption ionization-time of flight and the resistance profile was calculated using the disk diffusion agar technique. RESULTS: In total, 181 isolates were obtained, 93 from (surfaces), 18 from the air, and 70 clinical samples. S. epidermidis was the most frequent species (66.8%), and the failure rate in air cleaning was 100%. More than 60% of the isolates were MDR, and the majority of clinical isolates (60.4%) had a resistance profile identical to that of the environmental isolates. CONCLUSION: Staphylococcus spp. were found in most of the analyzed samples, with a high frequency of MDR isolates, demonstrating the importance of the hospital environment as a reservoir, and the need for infection control measures, and rational use of antimicrobials.

Essential Oils of Melaleuca, Citrus, Cupressus, and Litsea for the Management of Infections Caused by Candida Species: A Systematic Review.

Candida is a common agent of infection in humans, which has a wide distribution and is a colonizer fungus of the body, occasionally assuming the role of a pathogen. The type of treatment depends on the site of infection and the clinical condition of the patient. Superficial infections, such as mucosal infections, can be treated with topical medications. So-called alternative therapies have rarely been studied, although the literature records the effectiveness of some treatments, especially as complementary therapy. The aims of this review were to analyze evidence of the anti-Candida inhibitory activity of essential oils of the Citrus, Cupressus, Litsea, and Melaleuca species; in addition to addressing the chemical composition, probable mechanisms of antifungal action and studies of toxicity, cytotoxicity, and genotoxicity were included. The literature from Medline/PubMed, Science Direct, Scopus, Web of Science, and the Brazilian database Periodic Capes was reviewed. Thirty-eight articles were selected, which included two articles on Litsea spp., seven on Cupressus spp., thirteen articles on Citrus spp., and twenty-one articles on Melaleuca spp. In conclusion, this study showed in vitro evidence for the use of essential oils of the plant species evaluated for the treatment of infections caused by different Candida species.

Quantification of biofilm produced by clinical, environment and hands' isolates Klebsiella species using colorimetric and classical methods.

Some species of Klebsiella, such as Klebsiella pneumoniae and Klebsiella oxytoca, are important nosocomial pathogens frequently involved in outbreaks in Neonatal Intensive Care Units (NICU) and have the ability to form a biofilm. This study aims to evaluate the biofilm production of K. pneumoniae and K. oxytoca isolates collected from the hands of health professionals, neonates' blood and the environment of a Brazilian NICU, using three colorimetric methods and a classical method of counting the colony-forming units and compare the analysis among these techniques. The biofilm formation was carried out by the microplate technique, using three colorimetric assays: crystal violet, safranin and 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl) -5 [(phenylamino) arbonyl] - 2H-tetrazolium hydroxide (XTT). Also, colony-forming units were determined. Twenty-eight isolates of K. pneumoniae were collected from the blood, hands and environment and five of K. oxytoca from the hands and environment. All of them were strong biofilm producers, but K. pneumoniae isolates produced more biofilm than K. oxytoca when compared to the American Type Culture Collection (ATCC) strains used as positive controls. The number of viable cells in the biofilm produced by K. pneumoniae isolated from blood was significantly higher than in the control sample. Regarding the three colorimetric tests used in the study, the violet crystal obtained a higher absorbance average. The use of crystal-violet and XTT in the evaluation of biofilm in vitro make possible a complete analysis, since that it can quantify the total biomass (including the extracellular matrix) and evaluate the metabolic activity. In conclusion, this study identified isolates of K. pneumoniae and K. oxytoca that produce biofilms in the NICU and the bloodstream of neonates. This fact deserves attention since these patients are immunocompromised. The best methods will be chosen to answer research questions by always adopting more than one method so that more than one parameter or component of the biofilm is analyzed.

Plantas medicinais: uma abordagem sobre o uso seguro e racional / Medicinal plants: an approach to rational and safe use

Resumo Muitas plantas são utilizadas com finalidades medicinais, constituindo alternativas terapêuticas complementares ao tratamento de doenças, trazendo inúmeros benefícios à saúde, quando utilizadas racionalmente e de maneira adequada. No entanto, as plantas constituem um arsenal grande de constituintes químicos, que podem ser benéficos, mas também podem representar um risco potencial à saúde. Desse modo, é importante que o usuário, os profissionais de saúde, e os prescritores, tenham conhecimentos sobre a planta, a correta identificação, conservação, modo de preparo e uso, além dos possíveis efeitos colaterais. As plantas medicinais, seus riscos e benefícios, são discutidos à luz das publicações científicas contemporâneas, atentando para a contribuição dos profissionais de saúde em relação ao seu papel de educadores e promotores de saúde atuantes em comunidades, especialmente aquelas usuárias do Sistema Único de Saúde.

Abstract Many plants are used for medicinal purposes, constituting therapeutic alternatives complementary to the medical treatment, bringing numerous health benefits, when used rationally and in an appropriate way. However, plants constitute a large arsenal of chemicals constituents, which may be beneficial, but may also pose a potential health risk. So, it is important that the user, the health professionals, and the prescribers, have knowledge about the plant, ensuring correct identification, conservation, preparation and use, as well as possible side effects. The study of the medicinal plants, their risks and benefits, and efficacy confirmation by carefully designed studies will be discussed in the light of contemporary scientific publications, paying attention to the contribution of health professionals in relation to their role as active educators and health promoters in communities, especially those users of the Unified Health System.

Risk factors associated with mortality among patients who had candidemia in a university hospital.

INTRODUCTION: Bloodstream infection due to Candida spp. is a primary cause of morbidity and mortality in tertiary hospitals. METHODS: In this retrospective study, we included patients with a positive blood culture for Candida spp. after 48 h of hospitalization. RESULTS: A total of 335 patients who had candidemia were included in this study. Risk factors associated with mortality were hospitalization in internal medicine units and surgical clinics, age >60 years, mechanical ventilation, orotracheal intubation, hemodialysis, corticosteroids use, and C. parapsilosis infection. CONCLUSIONS: This study highlights the importance of health care related to invasive procedures and actions to improve patient immunity.

Onychomycosis in Psoriatic Patients with Nail Disorders: Aetiological Agents and Immunosuppressive Therapy.

Psoriasis and psoriatic arthritis are chronic, relapsing, immune-based diseases. Psoriatic patients may have nail involvement in 50 to 80% of cases, and this may reach 85% in patients with joint disease, in spite of the fact that the relationship between psoriasis and onychomycosis is not well established. The aim of this study was to investigate the occurrence of onychomycosis in patients with nail disorders and diagnosis of psoriasis and psoriatic arthritis. This was a cross-sectional study in which 38 patients diagnosed with psoriasis and/or psoriatic arthritis were interviewed and had altered nail samples analysed by mycological and histopathological exams. Twenty-two (57.89%) patients had a confirmed diagnosis for onychomycosis. Seventeen (44.8%) had a positive direct mycological examination, 16 (42.1%) had positive cultures, and 12 (31.6%) were positive for fungi by histopathological examination. Dermatophytes were identified in nine (56.3%) cultures, and of these, eight were Trichophyton rubrum and one T. tonsurans. Yeasts were isolated in seven patients (43.75%), which included four Candida parapsilosis and three C. albicans. Six patients (15.78%) were not using immunosuppressive therapy, and the others were using methotrexate, etanercept, adalimumab, infliximab, secukinumab, or golimumab, in monotherapy or in combination with other drugs. The confirmed onychomycosis rate in patients using methotrexate alone was 92.8% (n = 13). We concluded that it is possible that there is a positive relationship between psoriatic disease and onychomycosis. And we highlight that it is also worth investigating in the future the possible role of immunosuppressive therapy (mainly methotrexate) as a predisposing factor for the development of fungal infections in psoriatic patients.

Evaluation of Vulvovaginitis and Hygiene Habits of Women Attended in Primary Health Care Units of the Family.

BACKGROUND: This study aims to investigate the occurrence of vulvovaginal infections, to describe and to relate the hygiene habits of women who were treated at the Units of Primary Attention to Family Health in a municipality in the interior of Brazil. METHODS: This was a cross-sectional study that was carried out in two Primary Health Care Units of the Family of Uberlândia, Minas Gerais, Brazil. The population sample consisted of 100 women who underwent gynecological consultation in the health units. Data were collected through interviews and by the analysis of medical records. Results were reported according to laboratory reports. RESULTS: The participants' ages ranged from 18 to 45 years, with a median of 36 years. Thirty women (30%) presented alterations in the microbiota from a Pap smear, where bacterial vaginosis (diagnosed with Amsel criteria) was the predominant microorganism (83.3%). More than half of the women in the study who had vulvovaginitis wore cotton panties (70%, p = 0.651) and tight jeans/pants (83.3%, p = 0.010). The habit of depilating the genital area was reported by all the women, with the razor blade being the most commonly used tool (68%; p = 0.196). CONCLUSION: Bacterial vaginosis was the most commonly reported infection in the cytological exams. Care with genitalia and genital hygiene habits may be associated with the occurrence of vulvovaginitis.

Risk factors associated with mortality among patients who had candidemia in a university hospital

Abstract INTRODUCTION: Bloodstream infection due to Candida spp. is a primary cause of morbidity and mortality in tertiary hospitals. METHODS: In this retrospective study, we included patients with a positive blood culture for Candida spp. after 48 h of hospitalization. RESULTS A total of 335 patients who had candidemia were included in this study. Risk factors associated with mortality were hospitalization in internal medicine units and surgical clinics, age >60 years, mechanical ventilation, orotracheal intubation, hemodialysis, corticosteroids use, and C. parapsilosis infection. CONCLUSIONS: This study highlights the importance of health care related to invasive procedures and actions to improve patient immunity.

In Vitro and In Vivo Anti-Candida spp. Activity of Plant-Derived Products.

Candidiasis therapy, especially for candidiasis caused by Candida non-albicans species, is limited by the relatively reduced number of antifungal drugs and the emergence of antifungal tolerance. This study evaluates the anticandidal activity of 41 plant-derived products against Candida species, in both planktonic and biofilm cells. This study also evaluates the toxicity and the therapeutic action of the most active compounds by using the Caenorhabditis elegans-Candida model. The planktonic cells were cultured with various concentrations of the tested agents. The Cupressus sempervirens, Citrus limon, and Litsea cubeba essential oils as well as gallic acid were the most active anticandidal compounds. Candida cell re-growth after treatment with these agents for 48 h demonstrated that the L. cubeba essential oil and gallic acid displayed fungistatic activity, whereas the C. limon and C. sempervirens essential oils exhibited fungicidal activity. The C. sempervirens essential oil was not toxic and increased the survival of C. elegans worms infected with C. glabrata or C. orthopsilosis. All the plant-derived products assayed at 250 µg/mL affected C. krusei biofilms. The tested plant-derived products proved to be potential therapeutic agents against Candida, especially Candida non-albicans species, and should be considered when developing new anticandidal agents.

Isolation of Cryptococcus species from the external environments of hospital and academic areas.

INTRODUCTION: Fungi of the genus Cryptococcus are cosmopolitan and may be agents of opportunistic mycoses in immunocompromised and sometimes immunocompetent individuals. Cryptococcus species are frequently isolated from trees and bird excreta in the environment and infection occurs by inhalation of propagules dispersed in the air. The aim was to investigate Cryptococcus species in bird excreta and tree hollows located in a university hospital area and in an academic area of a university campus. METHODOLOGY: A total of 40 samples of bird excreta and 41 samples of tree hollows were collected. The identification of the isolates was done by classical methodology and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Twenty (62.5%) isolates of Cryptococcus were found in bird excreta and 12 (37.5%) in tree hollows. C. laurentii (currently Papiliotrema laurentii) was the most frequent species in both samples, being found in 5 samples of excreta and in 8 tree hollows. The diversity of species found in excreta (C. laurentii, C. albidus [currently Naganishia albida], C. liquefaciens [currently N. liquefaciens], C. friedmanii [currently N. friedmannii] and others) was higher than in tree hollows (C. laurentii, C. flavescens [currently Papiliotrema flavescens], and other yeasts). CONCLUSION: Many Cryptococcus species were isolated from excreta and tree hollows, and this fact is important for understanding the environmental epidemiology of those emerging pathogens for public health, as a way to implement surveillance actions and control of cryptococcosis.

Identificação, Prevenção e Tratamento da Síndrome Mão-Pé Induzida por Quimioterapia: Revisão Sistemática / Identification, Prevention and Treatment of Hand-Foot Syndrome Induced by Chemotherapy: Systematic Review / Identificación, Prevención y Tratamiento de la Enfermedad Mental por Quimioterapia: Revisión Sistemática

Introdução: A síndrome mão-pé é uma reação adversa experimentada por vários pacientes em tratamento para o câncer e fator preditor de morbidade e mortalidade. Objetivo: Avaliar as evidências científicas relacionadas à identificação, prevenção e tratamento da síndrome mão-pé induzida por agentes quimioterápicos, identificar os principais sinais e sintomas que possibilitam o reconhecimento da síndrome e, ainda, discutir a ocorrência de onicomicoses no contexto da síndrome mão-pé. Método: Trata-se de uma revisão sistemática na MEDLINE/PubMed, Biblioteca Virtual da Saúde e Scopus, incluindo literatura cinzenta e busca manual. Os 29 estudos incluídos na revisão foram analisados e classificados segundo a hierarquia dos níveis de evidência Grading of Recommendations Assessment, Development and Evaluations (GRADE) e a confiabilidade entre os examinadores (coeficiente Kappa) foi calculada. Resultados: Foram identificados estudos que demonstraram eficácia na prevenção da síndrome mão-pé com o uso da crioterapia e hidroterapia. Evidenciaram-se resultados satisfatórios com o uso do creme de ureia na prevenção e tratamento, e o uso de piridoxina não apresentou resultados conclusivos. Foram encontrados mecanismos para identificação da síndrome e para classificação dos agentes indutores. O grupo dos taxanos predominou entre os medicamentos indutores da síndrome mão-pé. Conclusão: Existem evidências consistentes, porém não contemplam todos os fármacos indutores da síndrome e não exploram outras manifestações relacionadas às onicólises e onicomicoses. O estudo apresentou resultados que poderão auxiliar os prescritores na identificação da síndrome mão-pé, além de alternativas para prevenção e tratamento. Contudo, vale destacar a necessidade de pesquisas futuras para elucidar a etiologia e protocolos de tratamento.

Introduction: Hand-foot syndrome is an adverse reaction experienced by many cancer patients and a predictor of morbidity and mortality. Objective:To evaluate the scientific evidence related to the identification, prevention and treatment of chemotherapeutic-induced hand-foot syndrome, to identify the main signs and symptoms that enable the recognition of the syndrome, and to discuss the occurrence of onychomycosis in the context of the hand-foot syndrome. Method: This is a systematic review at MEDLINE/PubMed, Virtual Health Library and Scopus, including gray literature and manual search. The 29 studies included in the review were analyzed and graded according to the hierarchy of evidence levels Grading of Recommendations Assessment, Development and Evaluations (GRADE) and reliability among examiners (Kappa coefficient) was calculated. Results:It were identified studies that demonstrated efficacy in preventing hand-foot syndrome using cryotherapy and hydrotherapy. Satisfactory results were evidenced with the use of urea cream for prevention and treatment, and the use of pyridoxine showed inconclusive results. Mechanisms for identification of the syndrome and classification of inducing agents were found. The taxane group predominated among hand-foot syndrome inducing drugs. Conclusion: There are consistent evidences but do not include all drugs inducing the syndrome and do not explore other manifestations related to onycholysis and onychomycosis. The study presented results that may help prescribers to identify hand-foot syndrome, as well as alternatives for prevention and treatment. However, it is worth highlighting the need for future studies to elucidate the etiology and treatment protocols.

Introducción: El síndrome de pies y manos es una reacción adversa experimentada por muchos pacientes con cáncer y un predictor de morbilidad y mortalidad. Objetivo: Evaluar la evidencia científica relacionada con la identificación, prevención y tratamiento del síndrome de pies y manos inducido por quimioterapia, identificar los principales signos y síntomas que permiten el reconocimiento del síndrome y analizar la aparición de onicomicosis en el contexto del síndrome mano-pie. Método:Esta es una revisión sistemática en MEDLINE/PubMed, Virtual Health Library y Scopus, que incluye literatura gris y búsqueda manual. Los 29 estudios incluidos en la revisión se analizaron y clasificaron de acuerdo con la jerarquía de los niveles de evidencia Grading of Recommendations Assessment, Development and Evaluations (GRADE). Resultados: Identificamos estudios que demostraron eficacia en la prevención del síndrome mano-pie usando crioterapia e hidroterapia. También mostraron resultados satisfactorios con el uso de crema de urea en la prevención y el tratamiento, y el uso de piridoxina no mostró resultados concluyentes. Se encontraron mecanismos para la identificación del síndrome y la clasificación de los agentes inductores. El grupo de taxanos predominó entre los fármacos inductores del síndrome mano-pie. Conclusión: Existe evidencia consistente pero no incluye todas las drogas que inducen el síndrome y no explora otras manifestaciones relacionadas con la onicólisis y la onicomicosis. El estudio presentó resultados que pueden ayudar a los prescriptores a identificar el síndrome de manos y pies, así como alternativas para la prevención y el tratamiento. Sin embargo, vale la pena destacar la necesidad de futuras investigaciones para dilucidar la etiología y los protocolos de tratamiento.

Impact of a medication therapy management service offered to patients in treatment of breast cancer

ABSTRACT Cancer has high morbidity and mortality rates related to medication use and produce a costly impact in health care. Thus, patients require constant monitoring and proper coordination of care between different professionals. This study aimed to evaluate the impact generated by a Medication Therapy Management service (MTM) offered to patients with breast cancer in use of polypharmacy. Observational, exploratory, descriptive and retrospective study of a MTM service that included 93 patients. Sociodemographic and clinical data related to pharmacotherapy and the processes associated with the systematization of the service were collected and analyzed. Patients were followed-up by the MTM service on average for 18 months (±4.31) and 185 drug-related problems (DRP) were identified, an average of two DRP per patient. Of these DRP, 48.11% were resolved and 49.73% were in the resolution process. The most common DRP were in the categories of Indication (37.84%), followed by Safety (23.78%). The safety category showed the highest resolution rate (59.09%). The study revealed an increased risk of DRP for patients with three or more comorbidities and using 5 or more medications. The process of systematization of a MTM service in oncology was associated with positive outcomes.

The epidemiology of cryptococcosis and the characterization of Cryptococcus neoformans isolated in a Brazilian University Hospital.

Cryptococcosis, a systemic disease caused by the fungus Cryptococcus neoformans/ Cryptococcus gattii is more severe in immunocompromised individuals. This study aimed to analyze the epidemiology of the disease, the molecular characteristics and the antifungal susceptibility of C. neoformans isolated from patients treated in a Brazilian university hospital. This retrospective study was conducted in the Clinical Hospital, Federal University of Uberlândia, and evaluated cases of cryptococcosis and strains of C. neoformans isolated from 2004 to 2013. We evaluated 41 patients, 85% of whom were diagnosed with AIDS. The fungus was isolated from the cerebrospinal fluid (CSF) of 21 patients (51%); 19.5% had fungemia and in 24% the agent was isolated from the CSF and blood, concurrently. Meningoencephalitis was the most frequent (75%) manifestation of infection. Despite adequate treatment, the mortality of the disease was 58.5%. Most isolates (97.5%) presented the VNI genotype (serotype A, var. grubii) and one isolate was genotyped as C. gattii (VGI); all the isolates were determined as mating type MATa and showed susceptibility to the tested antifungals (fluconazole, voriconazole, amphotericin B and 5-flucytosine). Although AIDS detection rates remain stable, opportunistic infections such as cryptococcosis remain as major causes of morbidity and mortality in these patients.

The epidemiology of cryptococcosis and the characterization of Cryptococcus neoformans isolated in a Brazilian University Hospital

ABSTRACT Cryptococcosis, a systemic disease caused by the fungus Cryptococcus neoformans/ Cryptococcus gattii is more severe in immunocompromised individuals. This study aimed to analyze the epidemiology of the disease, the molecular characteristics and the antifungal susceptibility of C. neoformans isolated from patients treated in a Brazilian university hospital. This retrospective study was conducted in the Clinical Hospital, Federal University of Uberlândia, and evaluated cases of cryptococcosis and strains of C. neoformans isolated from 2004 to 2013. We evaluated 41 patients, 85% of whom were diagnosed with AIDS. The fungus was isolated from the cerebrospinal fluid (CSF) of 21 patients (51%); 19.5% had fungemia and in 24% the agent was isolated from the CSF and blood, concurrently. Meningoencephalitis was the most frequent (75%) manifestation of infection. Despite adequate treatment, the mortality of the disease was 58.5%. Most isolates (97.5%) presented the VNI genotype (serotype A, var. grubii) and one isolate was genotyped as C. gattii (VGI); all the isolates were determined as mating type MATa and showed susceptibility to the tested antifungals (fluconazole, voriconazole, amphotericin B and 5-flucytosine). Although AIDS detection rates remain stable, opportunistic infections such as cryptococcosis remain as major causes of morbidity and mortality in these patients.

RELATED FACTORS FOR COLONIZATION BY Candida SPECIES IN THE ORAL CAVITY OF HIV-INFECTED INDIVIDUALS.

UNLABELLED: The colonization of the oral cavity is a prerequisite to the development of oropharyngeal candidiasis. AIMS: The aims of this study were: to evaluate colonization and quantify Candida spp. in the oral cavity; to determine the predisposing factors for colonization; and to correlate the levels of CD4+ cells and viral load with the yeast count of colony forming units per milliliter (CFU/mL) in HIV-positive individuals treated at a University Hospital. Saliva samples were collected from 147 HIV patients and were plated on Sabouraud Dextrose Agar (SDA) and chromogenic agar, and incubated at 30 ºC for 72 h. Colonies with similar morphology in both media were counted and the result expressed in CFU/mL. RESULTS: Of the 147 HIV patients, 89 had positive cultures for Candida spp., with a total of 111 isolates, of which C. albicans was the most frequent species (67.6%), and the mean of colonies counted was 8.8 × 10³ CFU/mL. The main predisposing factors for oral colonization by Candida spp. were the use of antibiotics and oral prostheses. The use of reverse transcriptase inhibitors appears to have a greater protective effect for colonization. A low CD4+ T lymphocyte count is associated with a higher density of yeast in the saliva of HIV patients.

RELATED FACTORS FOR COLONIZATION BY Candida SPECIES IN THE ORAL CAVITY OF HIV-INFECTED INDIVIDUALS / Fatores relacionados a colonização da cavidade bucal de indivíduos portadores do HIV por espécies de Candida

The colonization of the oral cavity is a prerequisite to the development of oropharyngeal candidiasis. Aims: The aims of this study were: to evaluate colonization and quantify Candida spp. in the oral cavity; to determine the predisposing factors for colonization; and to correlate the levels of CD4+ cells and viral load with the yeast count of colony forming units per milliliter (CFU/mL) in HIV-positive individuals treated at a University Hospital. Saliva samples were collected from 147 HIV patients and were plated on Sabouraud Dextrose Agar (SDA) and chromogenic agar, and incubated at 30 ºC for 72 h. Colonies with similar morphology in both media were counted and the result expressed in CFU/mL. Results: Of the 147 HIV patients, 89 had positive cultures for Candida spp., with a total of 111 isolates, of which C. albicans was the most frequent species (67.6%), and the mean of colonies counted was 8.8 × 10³ CFU/mL. The main predisposing factors for oral colonization by Candida spp. were the use of antibiotics and oral prostheses. The use of reverse transcriptase inhibitors appears to have a greater protective effect for colonization. A low CD4+ T lymphocyte count is associated with a higher density of yeast in the saliva of HIV patients.

RESUMO A colonização da cavidade oral pode ser considerada um pré-requisito para o desenvolvimento de candidíase orofaríngea. Os objetivos deste estudo foram: avaliar e quantificar espécies de Candidaisoladas da cavidade oral, para determinar os fatores predisponentes para a colonização, e correlacionar os níveis de células CD4+ e carga viral em indivíduos HIV-positivos atendidos em um hospital universitário. Foram coletadas amostras de saliva de 147 pacientes portadores do HIV, as quais foram semeadas em Ágar Sabouraud Dextrose (ASD) e ágar cromogênico e incubadas a 30 °C por 72 horas. As colônias com morfologia semelhante em ambos os meios foram contadas e o resultado expresso em unidade formadora de colônias por mililitro (UFC/mL). Dos 147 pacientes HIV positivos, 89 apresentaram culturas positivas para Candidaspp., totalizando 111 isolados, e C. albicansfoi a espécie mais frequente (67,6%). A contagem média de colônias foi de 8.8 × 10³ UFC/mL. Os principais fatores predisponentes para colonização oral por Candidaspp. foram a utilização de antibióticos e de próteses orais. O uso de antirretroviral da classe de inibidores da transcriptase reversa pareceu ter maior efeito protetor para a colonização. Baixa contagem de linfócitos T CD4+ está relacionada com maior densidade de leveduras na saliva de indivíduos HIV positivos.

FREQUENCY OF Candida SPECIES IN A TERTIARY CARE HOSPITAL IN TRIANGULO MINEIRO, MINAS GERAIS STATE, BRAZIL.

Infections by Candida species are a high-impact problem in public health due to their wide incidence in hospitalized patients. The goal of this study was to evaluate frequency, susceptibility to antifungals, and genetic polymorphism of Candida species isolated from clinical specimens of hospitalized patients. The Candida isolates included in this study were obtained from blood cultures, abdominal fluids, and central venous catheters (CVC) of hospitalized patients at the Clinical Hospital of the Federal University of Uberlândia during the period of July 2010 - June 2011. Susceptibility tests were conducted by the broth microdilution method. The RAPD-PCR tests used employed initiator oligonucleotides OPA09, OPB11, and OPE06. Of the 63 Candida isolates, 18 (28.5%) were C. albicans, 20 (31.7%) were C. parapsilosis complex species, 14 (22.2%) C. tropicalis, four (6.4%) C. glabrata, four (6.4%) C. krusei, two (3.3%) C. kefyr, and one (1.6%) C. lusitaniae. In vitro resistance to amphotericin B was observed in 12.7% of isolates. In vitro resistance to azoles was not detected, except for C. krusei. The two primers, OPA09 and OPB11, were able to distinguish different species. Isolates of C. albicans and C. parapsilosis complex species presented six and five clusters, respectively, with the OPA09 marker by RAPD-PCR, showing the genetic variability of the isolates of those species. It was concluded that members of the C. parapsilosis complex were the most frequent species found, and most isolates were susceptible to the antifungals amphotericin B, flucozanole, and itraconazole. High genetic polymorphisms were observed for isolates of C. albicans and C. parapsilosis complex species, mainly with the OPA09 marker.

FREQUENCY OF Candida SPECIES IN A TERTIARY CARE HOSPITAL IN TRIANGULO MINEIRO, MINAS GERAIS STATE, BRAZIL / Frequência de espécies de Candida em hospital terciário do Triângulo Mineiro, Minas Gerais, Brasil

Infections by Candida species are a high-impact problem in public health due to their wide incidence in hospitalized patients. The goal of this study was to evaluate frequency, susceptibility to antifungals, and genetic polymorphism of Candida species isolated from clinical specimens of hospitalized patients. The Candida isolates included in this study were obtained from blood cultures, abdominal fluids, and central venous catheters (CVC) of hospitalized patients at the Clinical Hospital of the Federal University of Uberlândia during the period of July 2010 - June 2011. Susceptibility tests were conducted by the broth microdilution method. The RAPD-PCR tests used employed initiator oligonucleotides OPA09, OPB11, and OPE06. Of the 63 Candida isolates, 18 (28.5%) were C. albicans, 20 (31.7%) were C. parapsilosis complex species, 14 (22.2%) C. tropicalis, four (6.4%) C. glabrata, four (6.4%) C. krusei, two (3.3%) C. kefyr, and one (1.6%) C. lusitaniae. In vitro resistance to amphotericin B was observed in 12.7% of isolates. In vitro resistance to azoles was not detected, except for C. krusei. The two primers, OPA09 and OPB11, were able to distinguish different species. Isolates of C. albicans and C. parapsilosis complex species presented six and five clusters, respectively, with the OPA09 marker by RAPD-PCR, showing the genetic variability of the isolates of those species. It was concluded that members of the C. parapsilosis complex were the most frequent species found, and most isolates were susceptible to the antifungals amphotericin B, flucozanole, and itraconazole. High genetic polymorphisms were observed for isolates of C. albicans and C. parapsilosis complex species, mainly with the OPA09 marker.

As infecções causadas por espécies de Candida são problema de grande impacto para a saúde pública, devido à alta incidência em pacientes hospitalizados e como causa de mortalidade. O presente estudo teve como objetivo avaliar a frequência de Candida spp. isoladas de pacientes hospitalizados, assim como a sensibilidade aos antifúngicos e o polimorfismo genético por RAPD-PCR. Os microrganismos incluíram isolados de hemocultura, líquido abdominal e ponta de cateter venoso central de pacientes internados no Hospital de Clínicas da Universidade Federal de Uberlândia, região do Triângulo Mineiro, Minas Gerais, Brasil, no período de julho de 2010-junho de 2011. Os testes de sensibilidade aos antifúngicos foram realizados por microdiluição em caldo e na análise por RAPD-PCR foram utilizados os oligonucleotídeos OPA09, OPB11, e OPE06. Dos 63 isolados, 18 (28,5%) foram C. albicans, 20 (31,7%) C. parapsilosis, 14 (22,2%) C. tropicalis, quatro (6,4%) C. glabrata, quatro (6,4%) C. krusei, dois (3,3%) C. kefyr, e um (1,6%) C. lusitaniae. Resistência in-vitro à anfotericina B foi observada em 12,7% dos isolados. Não foi observada resistência in-vitro aos azólicos, exceto para os isolados de C. krusei. Os oligonucleotídeos OPA09 e OPB11 possibilitaram distinguir diferentes espécies. Isolados de C. albicans apresentaram seis clusters e o complexo C. parapsilosis, cinco clusters, com o iniciador OPA09, por RAPD-PCR, mostrando a variabilidade genética daquelas espécies. Conclui-se que o complexo C. parapsilosis foi a espécie mais frequente, e a maioria dos isolados foi sensível in vitro aos antifúngicos testados. Alto polimorfismo genético foi observado para os isolados de C. albicans e complexo C. parapsilosis, principalmente com o oligonucleotídeo OPA09.

Enzymatic and hemolytic activity in different Candida species / La actividad enzimática y hemolítica en diferentes especies de Candida

Background: Candida species, in conditions of microbiota imbalance or decreased immune defenses, may be one of the main human fungal pathogens. Virulence factors constitute the mechanisms used by the fungus to avoid host defenses. Aims: This study aimed to investigate the in vitro production of virulence factors, such as hemolytic activity, and deoxyribonuclease (DNase), proteinase, and phospholipase activities in Candida spp. Methods: Fifty clinical isolates were analyzed for virulence factors: Candida albicans (15), Candida tropicalis (15), Candida parapsilosis (10), Candida glabrata (5), and Candida krusei (5). Hemolytic activity was determined in Sabouraud dextrose agar plates containing 3% glucose and 7% sheep red cells. Culture media containing, respectively, agar-base DNA, egg yolk, and bovine albumin were used to determine DNase, phospholipase and proteinase activities, respectively. Results: Forty-eight (96%) of 50 isolates showed hemolytic activity, with 10 (20%) positive for DNase, 19 (38%) for proteinase, and 16 (32%) for phospholipase. Statistically significant differences were observed between species for phospholipase (p < 0.0001) and proteinase (p < 0.05) production. Conclusions: It is concluded that all species had hemolytic activity. DNase activity was detected in all species except in C. glabrata; proteinase activity was detected in C. albicans, C. tropicalis, and C. parapsilosis; and phospholipase activity was observed in C. albicans and C. tropicalis (AU)

Antecedentes: Las levaduras del género Candida, en condiciones de desequilibrio de la microbiota o de disminución de las defensas inmunológicas, pueden ser uno de los principales patógenos fúngicos del hombre. Los factores de virulencia constituyen los mecanismos utilizados por el hongo para evadir las defensas del huésped. Objetivos: Este estudio tiene como objetivo investigar la producción in vitro de algunos factores de virulencia, como la actividad hemolítica, y las actividades desoxirribonucleasa (DNasa), proteinasa y fosfolipasa en Candidaspp. Métodos: Se analizaron 50 aislamientos clínicos: Candida albicans (15), Candida tropicalis (15), Candida parapsilosis (10), Candida glabrata (5), y Candida krusei (5). La actividad hemolítica fue determinada en placas de agar glucosado de Sabouraud, con glucosa al 3% y un 7% de hematíes de oveja. Los medios de cultivo de agar-ADN, yema de huevo y albúmina bovina fueron utilizados para determinar las actividades DNasa, fosfolipasa y proteinasa, respectivamente. Resultados: De los 50 aislamientos, 48 (96%) presentaron actividad hemolítica, 10 (20%) fueron positivos para DNasa, 19 (38%) para proteinasa y 16 (32%) para fosfolipasa. Se observaron diferencias estadísticamente significativas entre las especies para las actividades fosfolipasa (p < 0,0001) y proteasa (p < 0,05). Conclusiones: Se concluye que todas las especies estudiadas poseen actividad hemolítica. La actividad DNasa fue detectada en todas las especies, excepto en Candida glabrata; la actividad proteinasa fue detectada en C. albicans, C. tropicalis y C. parapsilosis, y la actividad fosfolipasa se observó en C. albicans y C. tropicalis (AU)