Secretion of five extracellular enzymes by strains of chromoblastomycosis agents

The gelatinase, urease, lipase, phospholipase and DNase activities of 11 chromoblastomycosis agents constituted by strains of Fonsecaea pedrosoi, F. compacta, Phialophora verrucosa, Cladosporium carrionii, Cladophialophora bantiana and Exophiala jeanselmei were analyzed and compared. All strains presented urease, gelatinase and lipase activity. Phospholipase activity was detected only on five of six strains of F. pedrosoi. DNase activity was not detected on the strains studied. Our results indicate that only phospholipase production, induced by egg yolk substrate, was useful for the differentiation of the taxonomically related species studied, based on their enzymatic profile.

Environmental contamination by Toxocara spp. Eggs in a rural settlement in Brazil

In order to study the environmental contamination by Toxocara spp. eggs in a rural community from the Pontal do Paranapanema region, São Paulo State, Brazil, soil samples from 31 out of 121 plots were collected in eight different places on each house. The samples were submitted to flotation technique in sodium nitrate (d = 1.20g/cm³). Eggs of Toxocara spp. were recovered in nine (29.03%) out of the 31 plots. At least one dog was registered in 27 of the 31 plots examined (87.1%) and at least one cat in 17 (54.84%). The number of pets per plot ranged from one to six (mean of 2.3) for dogs and one to 14 (mean of 1.29) for cats. In 16 plots (51.61%), the presence of both dogs and cats was observed. There was no relation between the presence of pets in the plots and soil contamination (p > 0.05). However, the environmental contamination by Toxocara spp. eggs associated to the poor conditions of the inhabitants may be an important risk factor for the human population to ocular or visceral larva migrans.

Molecular characterization of environmental Cryptococcus neoformans isolated in Vitoria, ES, Brazil

Cryptococcus neoformans is the major cause of fungal meningitis, a potentially lethal mycosis. Bird excreta can be considered a significant environmental reservoir of this species in urban areas, thirty-three samples of pigeon excreta were collected within the city of Vitoria, Brazil. Cryptococcus neoformans was isolated and identified using standard biochemical assays in ten samples. PCR amplification with primer M13 and orotidine monophosphate pyrophosphorylase (URA5) gene-restriction fragment length polymorphism (RFLP) analysis discerned serotypes and genotypes within this species. All isolates were serotype A (C. neoformans var. grubii) and genotype VNI. The two alternative alleles a and α at the mating type locus were determined by PCR amplification and mating assays performed on V8 medium. All isolates were MAT α mating type but only 50% were able to mate in vitro with the opposite mating type MAT a tester strains (JEC20, KN99a and Bt63). This study adds information on the ecology and molecular characterization of C. neoformans in the Southeast region of Brazil.

In vitro susceptibility testing of dermatophytes isolated in Goiania, Brazil, against five antifungal agents by broth microdilution method

The antifungal activities of fluconazole, itraconazole, ketoconazole, terbinafine and griseofulvin were tested by broth microdilution technique, against 60 dermatophytes isolated from nail or skin specimens from Goiania city patients, Brazil. In this study, the microtiter plates were incubated at 28 ºC allowing a reading of the minimal inhibitory concentration (MIC) after four days of incubation for Trichophyton mentagrophytes and five days for T. rubrum and Microsporum canis. Most of the dermatophytes had uniform patterns of susceptibility to the antifungal agents tested. Low MIC values as 0.03 µg/mL were found for 33.3%, 31.6% and 15% of isolates for itraconazole, ketoconazole and terbinafine, respectively.

Etiological agents of diarrhea in patients infected by the human immunodeficiency virus-1: a review

Despite the importance of understanding the epidemiology of agents responsible for infectious diarrhea in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) population, the number of articles about this subject is relatively few. The current article summarizes published data on bacterial, fungal, viral and parasitic enteropathogens in the HIV/AIDS seropositive subjects in different countries, regions and localities. In general, there is a great difference in the frequencies of etiological agents due to factors which include immune status, geographical location, climate and socioeconomic conditions. It is important to stress that a great prevalence of infection by emergent agents has been reported in the more advanced stages of AIDS. Therefore, to establish specific treatment depends directly on knowledge of these agents and risk factors associated to their distribution. Moreover, the colonization by potential pathogenic agents verified in these individuals is high thus implicating that they act as carriers. Finally, public health measures of control and prevention must take into consideration the regional previously identified enteropathogens, especially in areas where HIV prevalence is high.

Environmental analyses of the parasitic profile found in the sandy soil from the Santos municipality beaches, SP, Brazil

The environmental contamination by geohelminths represents a world public health problem and has been well documented by several authors. However, few papers describe the presence of such contamination in saline soils of coastal beaches. A study was performed on the beaches of the municipality of Santos in the period between May 2004 to April 2005 with the aim of determining the degree of contamination, and the correlation between contamination level and seasonal conditions and characteristics of the environment. Of the 2,520 samples analyzed, 18.2% (458) were contaminated, 32.3% (148) of which were localized in children's recreational areas (playgrounds). The parasite profile found in the analyzed samples indicated the presence of several zoonotic parasites: Ancylostoma larvae (82.5%), Toxocara sp. eggs (59.4%), Ancylostomidae-like eggs (37.1%), coccid oocysts (13.5%), Trichostrongylus sp. eggs and larvae, Ascaris lumbricoides eggs, (11.6%), Entamoeba sp. cysts (10.0%), Strongyloides sp. (4.8%), several free nematoids and some non-identified parasitic structures (3.3%). It was established that the highest frequency of parasitic structures occurred in the months between May and October 2004, and from February to March 2005. An increase in the diversity of parasitic forms was documented in the months between February to December 2004 and from January to April 2005, these periods having the highest rainfall.

Fungal agents in different anatomical sites in Public Health Services in Cuiabá, state of Mato Grosso, Brazil

INTRODUCTION: A contribution to the regional epidemiological profile of the most common fungal agents in Public Health Services in Cuiabá, state of Mato Grosso, including university hospitals and polyclinics. METHODS: Clinical specimens (n = 1,496) from 1,078 patients were collected, submitted to direct mycological exam (potash or stick tape method) and cultured in specific mediums. Dermatophytic and non-dermatophytic agents were identified according to micromorphology (Ridell technique). RESULTS: The majority of the 1,496 specimens were skin (n = 985) and nail exams (n = 472). Of the 800 positive cultures, 246 (30.8%) corresponded to dermatophytes and 336 (42%) to yeasts of the genus Candida, 190 (23.7%) to other yeasts, 27 (3.4%) to non-dermatophytic filamentous fungi and one (0.1%) the agent of subcutaneous mycosis. Lesions considered primary occurred in greater numbers (59.5%) than recurrent lesions (37.4%), with a greater concentration of positivity occurring on the arms and legs. CONCLUSIONS: Comorbidities, allergies and diabetes mellitus were conditions associated with greater positivity in direct mycological exams and cultures. Positive culture was considered a definitive diagnosis of fungal infection and confirmed 47.8% of diagnostic hypotheses.

Intestinal parasitism and socio-environmental factors among Mbyá-Guarani indians, Porto Alegre, Rio Grande do Sul, Brazil

Disturbing data reveal the prevalence of intestinal parasites and their relationship with socio-environmental factors among Mbyá-Guarani Indians. The prevalence was determined by spontaneous sedimentation in water, centrifugation-floatation, and Kato-Katz. A socioeconomic questionnaire was submitted to each family. The overall prevalence of intestinal parasites was 88.7%, and 45.5% were polyparasitized. There was 90.5% prevalence of enteric parasites in children (1-12- year-old), and 85% among 13-65-year-old individuals, indicating that both age groups are extensively parasitized. The parasite load was low to moderate for geohelminths and 75% of the families did not have latrine, thus the practice of defecation occurred outdoors. These findings suggest that the multiple intestinal parasitism in the Mbyá-Guarani community is high to the point of being the rule, and that it relates essentially to the traditional lifestyle and health habits. It is urgently necessary to implement the association of anti-parasitic treatment with sanitation improvement. This should be done simultaneously with health education activities for this population.

Contamination of public parks and squares from Guarulhos (São Paulo State, Brazil ) by Toxocara spp. and Ancylostoma spp.

The contaminated soil with mammal feces is an important factor of risk to infection with zoonotic diseases. Amongst these zoonoses are visceral larva migrans and cutaneous larva migrans caused by Toxocara spp. and Ancylostoma spp., respectively. The aim of this study was to assess the environmental contamination by Toxocara spp. eggs and hookworms (Ancylostoma spp.) in public parks and squares in the city of Guarulhos, a metropolitan area of São Paulo, São Paulo State, Brazil. Soil samples were collected, between September and December 2010, and examined using the centrifugal flotation technique with sodium dichromate and zinc sulphate as well as the modified Baermann method. Notably, 35 (74.5%) of the 47 districts surveyed in Guarulhos possessed samples contaminated with Toxocara spp. and/or eggs or larvae of Ancylostoma spp. The frequency of Toxocara spp. and Ancylostoma spp. in the samples from public areas was 68.1% and 46.8%, respectively. Overall, the eastern side of Guarulhos is the region with the highest occurrence of causative agents of larva migrans. In all collection sites, the presence of feces from dogs and cats accompanied by their owners and stray animals were observed. Notably, it is important to adopt measures to control dog and cat breeding, to treat infected animals, and provide health education to the population.

Genetic Control of Mosquitoes: population suppression strategies

Over the last two decades, morbidity and mortality from malaria and dengue fever among other pathogens are an increasing Public Health problem. The increase in the geographic distribution of vectors is accompanied by the emergence of viruses and diseases in new areas. There are insufficient specific therapeutic drugs available and there are no reliable vaccines for malaria or dengue, although some progress has been achieved, there is still a long way between its development and actual field use. Most mosquito control measures have failed to achieve their goals, mostly because of the mosquito's great reproductive capacity and genomic flexibility. Chemical control is increasingly restricted due to potential human toxicity, mortality in no target organisms, insecticide resistance, and other environmental impacts. Other strategies for mosquito control are desperately needed. The Sterile Insect Technique (SIT) is a species-specific and environmentally benign method for insect population suppression, it is based on mass rearing, radiation mediated sterilization, and release of a large number of male insects. Releasing of Insects carrying a dominant lethal gene (RIDL) offers a solution to many of the drawbacks of traditional SIT that have limited its application in mosquitoes while maintaining its environmentally friendly and species-specific utility. The self-limiting nature of sterile mosquitoes tends to make the issues related to field use of these somewhat less challenging than for self-spreading systems characteristic of population replacement strategies. They also are closer to field use, so might be appropriate to consider first. The prospect of genetic control methods against mosquito vectored human diseases is rapidly becoming a reality, many decisions will need to be made on a national, regional and international level regarding the biosafety, social, cultural and ethical aspects of the use and deployment of these vector control methods.

DETECTION OF VIRULENCE GENES IN ENVIRONMENTAL STRAINS OF Vibrio cholerae FROM ESTUARIES IN NORTHEASTERN BRAZIL

The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

Sporothrix schenckii COMPLEX: SUSCEPTIBILITIES TO COMBINED ANTIFUNGAL AGENTS AND CHARACTERIZATION OF ENZYMATIC PROFILES

SUMMARY Sporothrix schenckiiwas reclassified as a complex encompassing six cryptic species, which calls for the reassessment of clinical and epidemiological data of these new species. We evaluated the susceptibility of Sporothrix albicans (n = 1) , S. brasiliensis (n = 6) , S. globosa (n = 1), S. mexicana(n = 1) and S. schenckii(n = 36) to terbinafine (TRB) alone and in combination with itraconazole (ITZ), ketoconazole (KTZ), and voriconazole (VRZ) by a checkerboard microdilution method and determined the enzymatic profile of these species with the API-ZYM kit. Most interactions were additive (27.5%, 32.5% and 5%) or indifferent (70%, 50% and 52.5%) for TRB+KTZ, TRB+ITZ and TRB+VRZ, respectively. Antagonisms were observed in 42.5% of isolates for the TRB+VRZ combination. Based on enzymatic profiling, the Sporothrix schenckii strains were categorized into 14 biotypes. Leucine arylamidase (LA) activity was observed only for S. albicans and S. mexicana. The species S. globosaand S. mexicanawere the only species without β-glucosidase (GS) activity. Our results may contribute to a better understanding of virulence and resistance among species of the genus Sporothrixin further studies.

ISOLATION OF Cryptococcus neoformans FROM ENVIRONMENTAL SAMPLES COLLECTED IN SOUTHEASTERN NIGERIA

SUMMARY Cryptococcosis caused by Cryptococcus neoformans is the second most common fungal opportunistic pathogen and a lifethreatening infection with serious clinical manifestations especially in HIV/AIDS and other immunocompromised patients. In Nigeria, HIV/AIDS infection has reached an alarming level. Despite this, information on the presence of this fungus in clinical and environmental samples is very scanty in Nigeria and many other parts of Africa. We set out to evaluate the presence of Cryptococcus neoformans or C. gattii in pigeon droppings obtained from Southeastern Nigeria. One hundred and seventy-seven samples of pigeon droppings from six sample types were collected. The area covered comprised of ten cities and other locations spanning across five States in Nigeria. Using established techniques, Cryptococcus neoformans was isolated from 39 of the 177 (22.0%) samples overall. No C. gattiiwas isolated. Most of the isolates (32.4%) were recovered from dovecotes (11 of 34) followed closely by samples taken from markets (31.8%; seven of 22) and least from the church (4.0%; one of 25). The highest isolation rate (38.9%) was found in samples from Enugu-Ezike(seven of 23) while the least came from Afikpoand the other locations each with 9.1% isolation rate. This is the first large-scale screening of Cryptococcus neoformans from pigeon droppings in Nigeria. The ecological and epidemiological significance of these findings are discussed.

QUANTITATIVE VS. CONVENTIONAL PCR FOR DETECTION OF HUMAN ADENOVIRUSES IN WATER AND SEDIMENT SAMPLES

SUMMARY Human Adenoviruses (HAdV) are notably resistant in the environment. These agents may serve as effective indicators of fecal contamination, and may act as causative agents of a number of different diseases in human beings. Conventional polymerase chain reaction (PCR) and, more recently, quantitative PCR (qPCR) are widely used for detection of viral agents in environmental matrices. In the present study PCR and SYBR(r)Green qPCR assays were compared for detection of HAdV in water (55) and sediments (20) samples of spring and artesian wells, ponds and streams, collected from dairy farms. By the quantitative methodology HAdV were detected in 87.3% of the water samples and 80% of the sediments, while by the conventional PCR 47.3% and 35% were detected in water samples and sediments, respectively.