Reginaldo dos Santos Pedroso

Prevalence of intestinal parasites in preschool children in the region of Uberlândia, State of Minas Gerais, Brazil

INTRODUCTION Children are an important high-risk group for helminth and protozoa infections. Daycare centers are environments where children have proven to be more susceptible to acquiring intestinal parasites. Thus, the purpose of this study was to verify the prevalence of intestinal parasites in children who attended the two daycare centers maintained by the local government of Uberlândia, State of Minas Gerais, Brazil. METHODS Fecal samples were collected from 133 children (73 children at the Public Preschool for Early Childhood Education, PPECE A, and 60 at the PPECE B) following identification according to sex and age and agreement to participate by parents or guardians who signed the free, informed consent form. The samples were examined by the Lutz method. RESULTS Coproparasitological tests performed on 133 children showed that 29.3% of them were parasitized for enteroparasites or commensals, 6.7% of the children presented polyparasitism. Among the protozoa, Giardia lamblia were the most prevalent and Hymenolepis nana were the most frequent among the helminths. CONCLUSIONS Thus, analysis of the results showed that intestinal parasites still represent a public health problem, especially among children and in areas where the socioeconomic and educational conditions are less favorable.

In vitro susceptibility to antifungal agents of environmental Cryptococcus spp. isolated in the city of Ribeirão Preto, São Paulo, Brazil

Infections by Cryptococcus strains other than C. neoformans have been detected in immunocompromised patients. Of these strains, three are considered human pathogens: C. albidus, C. laurenttii, and C. uniguttulatus. This study deals with the in vitro susceptibility of Cryptococcus to drugs such as amphotericin B, itraconazole, fluconazole, and 5-fluorocytosine. Environmental Cryptococcus isolates (50) distributed as follows: C. neoformans var. neoformans (16), C. albidus (17), C. laurentii (14), and C. uniguttulatus (3) were evaluated by the micro and macrodilution techniques, according to EUCAST and NCCLS recommendations, respectively. Considering both methodologies the respective minimal inhibitory concentrations (MIC) were 0.125 and 2 microg/ml for amphotericin B, 0.06 and 8 microg/ml for itraconazole, and 0.5 and more than 64 microg/ml for fluconazole and 5-fluorocytosine. Agreement percentages for the two methodologies were 100% for amphotericin B and fluconazole for all the strains tested. For itraconazole, the agreement percentage was 81.3% in the C. neoformans strain and 100% for all the others. All species had a agreement percentage of 94.1 to 100% when susceptibility to 5-fluorocytosine was tested. It is concluded that environmental isolates of C. neoformans var. neoformans, C. albidus, C. laurentii, and C. uniguttulatus may show high MICs against certain drugs, suggesting in vitro primary resistance to the antifungals tested.

Avaliação da produção de melanina por espécies de Cryptococcus em quatro diferentes meios de cultura

The capacity of Cryptococcus spp to produce melanin in media containing phenol compounds is widely used for identifying these species in the laboratory. The aim of the present study was to compare the production of this pigment by Cryptococcus spp. in four culture media. Sixteen strains of Cryptococcus neoformans, 17 of Cryptococcus albidus, 13 of Cryptococcus laurentii and two of Cryptococcus uniguttulatus were tested in the following media: potato-carrot agar, Niger seed agar, sunflower seed agar and L-dopa agar. The melanin production was evaluated on the basis of colony pigmentation. Its production after five days of incubation was demonstrated by 93.8% of the strains of Cryptococcus neoformans in the media of potato-carrot agar, sunflower seed agar and L-dopa agar. From the isolates of Cryptococcus albidus, 29.4% produced the pigment in potato-carrot agar and L-dopa agar, 11.8% in Niger seed agar and 36% in sunflower seed agar. From Cryptococcus laurentii, 53.8% produced the pigment in potato-carrot agar and sunflower seed agar, 61.5% in L-dopa agar and 84.6% in Niger seed agar. Only one strain of Cryptococcus uniguttulatus presented slight production of the pigment, in potato-carrot agar.

Dermatophyte susceptibilities to antifungal azole agents tested in vitro by broth macro and microdilution methods

The in vitro susceptibility of dermatophytes to the azole antifungals itraconazole, fluconazole and ketoconazole was evaluated by broth macro and microdilution methods, according to recommendations of the CLSI, with some adaptations. Twenty nail and skin clinical isolates, four of Trichophyton mentagrophytes and 16 of T. rubrum were selected for the tests. Itraconazole minimal inhibitory concentrations (MIC) varied from < 0.03 to 0.25 microg/mL in the macrodilution and from < 0.03 to 0.5 microg/mL in the microdilution methods; for fluconazole, MICs were in the ranges of 0.5 to 64 microg/mL and 0.125 to 16 microg/mL by the macro and microdilution methods, respectively, and from < 0.03 to 0.5 microg/mL by both methods for ketoconazole. Levels of agreement between the two methods (+/- one dilution) were 70% for itraconazole, 45% for fluconazole and 85% for ketoconazole. It is concluded that the strains selected were inhibited by relatively low concentrations of the antifungals tested and that the two methodologies are in good agreement especially for itraconazole and ketoconazole.

Variability in UVB Tolerances of Melanized and Nonmelanized Cells of Cryptococcus neoformans and C. laurentii

Solar radiation is one of the major factors responsible for the control of fungus populations in the environment. Inactivation by UVA and UVB radiation is especially important for the control of fungi that disperse infective units through the air, including fungi such as Cryptococcus spp. that infect their vertebrate hosts by inhalation. Cryptococcus neoformans produces melanin in the presence of certain exogenous substrates such as l‐3,4 dihydroxyphenylalanine and melanization may protect the fungus against biotic and abiotic environmental factors. In the present study, we investigated the effect of exposure to an UVB irradiance of 1000 mW m−2 (biologically effective weighted irradiance) on the survival of melanized and nonmelanized cells of four strains of C. neoformans and four strains of C. laurentii. The relative survival (survival of cells exposed to radiation in relation to cells not exposed) of cells grown 2, 4, 6 or 8 days on medium with or without L‐dopa was determined after exposure to UVB doses of 1.8 and 3.6 kJ m−2. Both the irradiance spectrum and the intensities of those doses are environmentally realistic, and, in fact, occur routinely during summer months in temperate regions. Differences in tolerance to UVB radiation were observed between the C. neoformans and C. laurentii strains. The C. neoformans strains were more susceptible to UVB radiation than the C. laurentii strains. In C. neoformans, differences in tolerance to radiation were observed during development of both melanized and nonmelanized cells. For most treatments (strain, time of growth and UVB dose), there were virtually no differences in tolerances between melanized and nonmelanized cells, but when differences occurred they were smaller than those previously observed with UVC. In tests with two strains of C. laurentii, there was no difference in tolerance to UVB radiation between melanized and nonmelanized cells during 8 days of culture; and in tests with four strains for less culture time (4 days) there were no significant differences in tolerance between melanized and nonmelanized cells of any strain of this species.

Evaluation of virulence factors of Candida albicans isolated from HIV-positive individuals using HAART.

The colonization by Candida species is one of the most important factors related to the development of oral candidiasis in HIV-infected individuals. The aim of the study was to evaluate and discuss the phospholipase, proteinase, DNAse and haemolytic activities of Candida albicans isolated from the oral cavity of HIV individuals with high efficiency antiretroviral therapy. Seventy-five isolates of C. albicans obtained from saliva samples of patients with HIV and 41 isolates from HIV-negative individuals were studied. Haemolytic activity was determined in Sabouraud dextrose agar plates containing 3% glucose and 7% sheep red cells. Culture medium containing DNA base-agar, egg yolk, and bovine albumin were used to determine DNase, phospholipase and proteinase activities, respectively. All isolates from the HIV patients group had haemolytic activity, 98% showed phospholipase activity, 92% were positive for proteinase and 32% DNAse activity. Regarding the group of indivídios HIV negative, all 41 isolates presented hemolytic activity, 90.2% showed phospholipase and proteinase activity and 12.2% were positive for DNAse. The phospholipase activity was more intense for the group of HIV positive individuals. DNase production was more frequently observed in the group of HIV-positive individuals. The percentage of isolates having DNAse activity was also significantly different between the groups of patients not using any antiretroviral therapy, those using transcriptase inhibitors and those using transcriptase inhibitor and protease inhibitor in combination.

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 vitroresistance 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.

Phenotypic Characteristics of Yeasts of the Genus Candida and Cryptococcus in Differential Culture Media

In the clinical mycology laboratory, the identification of yeast species is done by screening in specific media, such as chromogenic agar for Candida species and Niger seed agar for Cryptococcus species, both of which are of clinical interest. This study aimed to evaluate the growth and morphological characteristics of yeasts of the Candida and Cryptococcus species in different culture media. Yeast species included in the study were: C. albicans, C. dubliniensis, C. glabrata, C. tropicalis, C. krusei, C. lipolytica, C. parapsilosis, C. metapsilosis, C. orthopsilosis, C. neoformans, C. gattii, C. flavescens, and C. albidus. The media used were Sabouraud dextrose agar, Sabouraud dextrose broth, hypertonic Sabouraud broth (plus 6.5% NaCl), Candida chromogenic agar, methyldopa agar, Niger seed agar and tobacco agar. Growth, color, size, presence of fringes, melanin and the appearance of the colony were evaluated. All isolates grew in the media used, except for the hypertonic Sabouraud broth; in Candida chromogenic agar, C. albicans and C. dubliniensis present a green color and C. tropicalis a blue color, while other species show colors including pink, purple, gray and white; in the Niger seed agar, C. neoformans, C. gattii and C. flavescens presented a brown color, while others had white colonies; in tobacco agar, the colors included white, cream and gray; and in methyldopa agar, all colonies were white. Some isolates presented colonies with fringes in the tobacco, methyldopa and Niger seed agar; the presence of melanin was observed by Cryptococcus isolates in the Niger seed and tobacco agar; the appearance of colonies in the media varied from opaque to shiny or mucoid, according to the isolate and the culture medium. All of the culture media used allowed the growth of the tested isolates, except for C. lipolytica, which did not grow in hypertonic Sabouraud broth. The isolates of Cryptococcus, C. krusei and C. dubliniensis presented a significant reduction of growth in hypertonic Sabouraud broth. K e y w o r d s

Alternative and complementary therapies for vulvovaginal candidiasis

When it comes to women’s health, treating vaginal infections makes up a high proportion of the gynecological services. Among the forms of vaginitis, vulvovaginal candidiasis (VVC) is considered the second most common. Demand for new treatment alternatives is increasingly relevant, especially for therapies with fewer side effects, better tolerability, and lower cost, while still offering improved quality of life in terms of disease prevention. This study intended to investigate the alternative therapies described for the adjuvant treatment of vulvovaginitis caused by Candida species, including alternative and complementary treatment methods used by women. This literature review is based on articles written in English and Portuguese in the PubMed, Google Scholar, and SciELO databases. This study was conducted for the most part using the Brazilian Government’s Capes Periodicals Portal, which directs to Google Scholar and PubMed. Since the 1980s, there has been growing interest in alternative therapies in Brazil, a trend which also began in other Western countries in the second half of the twentieth century. Some alternative treatments include substances with antifungal activity, some substances help restore the balance of the vaginal microbiota, while others have an inhibitory activity on microbial virulence factors. The proper use of therapeutic alternatives can effectively contribute to the treatment of VVC, but it should be remembered that some chemical products, such as boric acid or vinegar, and even natural products such as propolis, garlic, and tea tree may have undesirable side effects, having not been tested by well-designed clinical studies. Even so, alternative therapies in the treatment of VVC do have support in the scientific literature.

Bloodstream Infection: The Influence of Risk Factors, Etiology and Antimicrobial Therapy on Mortality Rates

Introduction: Inappropriate initial antimicrobial therapy leads to higher mortality in patients with bloodstream infection. This study aimed to evaluate the relationship between risk factors, etiology and antimicrobial therapy on mortality rates of patients with bloodstream infection. Methods: Between January 2016 to December 2016, 167 patients with bloodstream infection were prospectively evaluated according to the presence or absence of inappropriate antimicrobial therapy of infection. Hospital mortality was the main outcome variable compared between the two study groups. Results: Infected patients who received inappropriate antimicrobial therapy had statistically more diabetes mellitus, chronic obstructive pulmonary disease, chronic renal disease and death than infected patients who initially received appropriate antimicrobial therapy. Loading dose error and error in starting antimicrobial administration were the most frequently detected error in our study and both were determinant factors related to increased mortality. Initial antimicrobial therapy was maintained, escalation and de-escalation 67.6%, 22.7% and 9.6% of cases, respectively. Coagulase negative staphylococci represented the majority reaching 40.7% and multi-drug resistant microorganisms were detected in 27.3% of infections. There was no observed difference in mortality rates among infections caused by resistant or susceptible microorganisms. Conclusion: Loading dose error and error in starting antimicrobial administration, were the most frequently detected error in our study and both were determinant factors related to increased mortality. Beside the multiple logistic regression analysis revealed that the delay in starting antimicrobial therapy was the only independent factor that increased mortality.