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1.
Rev. Soc. Bras. Med. Trop ; 53: e20190336, 2020. tab, graf
Article in English | LILACS | ID: biblio-1057282

ABSTRACT

Abstract INTRODUCTION: Candida parapsilosis complex species differ from each other with regard to their prevalence and virulence. METHODS: The hydrolytic enzyme activity, biofilm production, and adhesion to epithelial cells were analyzed in 87 C. parapsilosis complex strains. RESULTS: Among the studied isolates, 97.7%, 63.2%, and 82.8% exhibited very strong proteinase, esterase, and hemolysin activity, respectively. All the C. parapsilosis complex isolates produced biofilms and presented an average adherence of 96.0 yeasts/100 epithelial cells. CONCLUSIONS: Our results show that Candida parapsilosis complex isolates showed different levels of enzyme activity, biofilm production, and adhesion to epithelial cells.


Subject(s)
Humans , Virulence Factors/analysis , Candida parapsilosis/pathogenicity , Cell Adhesion , Mycological Typing Techniques , Biofilms/growth & development , Candida parapsilosis/isolation & purification , Candida parapsilosis/classification , Candida parapsilosis/enzymology , Hydrolases/biosynthesis
2.
Clin. biomed. res ; 40(4): 213-217, 2020. tab
Article in Portuguese | LILACS | ID: biblio-1252521

ABSTRACT

Introdução: O método clássico para o diagnóstico de micoses é realizado pelo Exame Micológico Direto (EMD) e cultural, que possibilita a visualização de estruturas fúngicas vegetativas e estruturas reprodutivas, respectivamente. Essa combinação é fundamental para reduzir possíveis erros analíticos e aumentar a precisão do diagnóstico. Métodos: Com a finalidade de verificar a frequência do EMD e cultural, e comparar seus parâmetros de sensibilidade e especificidade, realizamos uma análise retrospectiva entre janeiro de 2018 e maio de 2020, de 1603 laudos micológicos oriundos de um laboratório de análises clínicas, localizado em Porto Alegre. Resultados: Após a análise dos laudos observamos que a maioria dos casos apresentaram o EMD negativo com cultura positiva (36,24%). Na sequência, 30,87% dos casos foram de amostras negativas e 25,57% dos laudos foram positivos para ambos os exames. A minoria dos casos (7,29%) apresentou o EMD positivo com cultura negativa. Conclusão: Esta análise revelou que o exame cultural é mais sensível e específico, demonstrando uma maior confiabilidade no diagnóstico. Entretanto, vale ressaltar que a realização dos exames em conjunto, além de reduzir possíveis erros analíticos, proporciona um diagnóstico melhor fundamentado. (AU)


Introduction: The classic method for the diagnosis of mycoses is performed by both direct mycological examination (DME) and culture, which allow the visualization of vegetative and reproductive fungal structures, respectively. This combination is essential to reduce possible analytical errors and increase the accuracy of the diagnosis. Methods: To assess the frequency of DME and culture, and compare their parameters of sensitivity and specificity, we performed a retrospective analysis of 1603 mycological reports produced between January 2018 and May 2020 in a clinical analysis laboratory in Porto Alegre, southern Brazil. Results: After analyzing the reports, we observed that most cases presented a negative DME and a positive culture (36.24%). Subsequently, 30.87% of the cases were negative for both tests, and 25.57% were positive for both tests. The minority of cases (7.29%) presented a positive DME and a negative culture. Conclusion: Our analysis revealed that cultural examination is more sensitive and specific, showing greater reliability in the diagnosis. However, it is noteworthy that performing the tests together, in addition to reducing possible analytical errors, provides a more consistent diagnosis. (AU)


Subject(s)
Comparative Study , Culture Media , Laboratory Test , Mycoses/diagnosis , Sensitivity and Specificity , Mycological Typing Techniques
3.
Rev. Soc. Bras. Med. Trop ; 53: e20190422, 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1136889

ABSTRACT

Abstract INTRODUCTION: The average annual incidence of cryptococcosis in Colombia is 0.23 cases per 100,000 inhabitants in the general population, and 1.1 cases per 1000 in inhabitants with Acquired Immune Deficiency Syndrome (AIDS). In addition, the causal fungus has been isolated from the environment, with serotypes A-B and C in different regions. This study aims to determine the genetic association between clinical and environmental isolates of C. neoformans/C. gattii in Colombia. METHODS: Multilocus sequence typing (MLST) was used to identify possible clones, providing information about the epidemiology, ecology, and etiology of this pathogen in Colombia. RESULTS: A total of 110 strains, both clinical (n=61) and environmental (n=49), with 21 MLST sequence types (ST) of C. neoformans (n=14STs) and C. gattii (n=7STs) were identified. The STs which shared clinical and environmental isolate sources were grouped in different geographical categories; for C. neoformans, ST93 was identified in six departments, ST77 in five departments; and for C. gattii, ST25 was identified in three departments and ST79 in two. CONCLUSIONS: High genetic diversity was found in isolates of C. neoformans/gattii by MLST, suggesting the presence of environmental sources harboring strains which may be sources of infection for humans, especially in immunocompromised patients; these data contribute to the information available in the country on the distribution and molecular variability of C. neoformans and C. gattii isolates recovered in Colombia.


Subject(s)
Humans , Cryptococcosis , Cryptococcus neoformans , Cryptococcus gattii , Genetic Variation , Mycological Typing Techniques , Colombia , Multilocus Sequence Typing , Genotype
4.
Article in French | AIM, AIM | ID: biblio-1264286

ABSTRACT

Introduction : Les teignes sont des affections fongiques contagieuses causées par plusieurs espèces de dermatophytes. Cette mycose touche essentiellement l'enfant et rarement l'adulte. Les teignes anthropophiles sont fréquentes dans la plupart des pays en voie de développement. Cette affection reste sous-documentée à Madagascar. Notre étude a pour objectif de rapporter les cas de teignes diagnostiqués dans le laboratoire de parasitologie-mycologie au CHU Ravoahangy Andrianavalona Antanananarivo de 2005 à 2018. Méthode : Il s'agit d'une étude rétrospective descriptive incluant tous les dossiers des patients ayant effectué un examen mycologique. Ont été inclus les dossiers comportant comme diagnostic la teigne. Chacun de ces patients a bénéficié d'un examen direct et d'une culture mycologique Résultats : Nous avons colligé en 13 ans 1014 patients confirmés porteurs de mycose. La fréquence des teignes sur l'ensemble des mycoses a été de 5,81% (59/1014). La prévalence brute des teignes a été de 37,57% (59/157). L'âge des patients variait de 2 à 67 ans dont 52,54% sont des enfants moins de 10 ans. La moyenne d'âge est de 13,5 ans. Les teignes étaient plus retrouvées chez les hommes (71,19 %) que chez les femmes (28,81 %) avec un sex ratio H/F de 2,47.Parmi ces patients, 77, 96% ont eu une notion de traitement avant l'examen mycologique. Dix souches de dermatophytes ont été isolées. Parmi les espèces retrouvées, Microsporium langeronii est l'espèce la plus isolée (33,89 %), suivie de Trichophyton mentagrophytes à 20,33 %. Conclusion : La fréquence des teignes n'est pas négligeable à Madagascar atteignant préférentiellement les enfants. Le diagnostic biologique des teignes est indispensable avant de débuter le traitement. L'identification de l'agent causal est importante pour prévenir et contrôler l'infection dermatophytique


Subject(s)
Madagascar , Mycological Typing Techniques , Scalp , Tinea/diagnosis , Tinea/epidemiology , Tinea/etiology
5.
Braz. j. infect. dis ; 22(6): 495-498, Nov.-Dec. 2018. tab
Article in English | LILACS | ID: biblio-1039219

ABSTRACT

ABSTRACT There are limited data on the molecular epidemiology of cryptococcosis in Brazil. Here, we report on the identification of the molecular pattern of the Cryptococcus species that caused meningitis in patients admitted in a Brazilian reference tertiary care hospital, and review the published studies addressing the molecular epidemiology of Cryptococcus in Brazil. Our study has shown the predominance of molecular type VNII in HIV-infected patients with cryptococcal meningoencephalitis. Molecular types VNII and VGII were occasionally detected in HIV-infected and non-infected patients with meningoencephalitis. In contrast, previous studies have shown that several regions exhibited a high prevalence of the VNI molecular type and sporadic cases of the VNII and VGII molecular types in patients with cryptococcosis in Brazil. Additional studies including VNII isolates will contribute to understanding the epidemiology and phylogenetic relationship of these genotype compared to the other ones. So far, no clear correlation has been established between genotypes, antifungal susceptibility for Cryptococcus and clinical outcome in cryptococcosis.


Subject(s)
Humans , Male , Female , Infant , Adult , Middle Aged , Aged , Meningitis, Cryptococcal/microbiology , Cryptococcus neoformans/genetics , Polymorphism, Restriction Fragment Length , Brazil/epidemiology , HIV Infections/microbiology , Mycological Typing Techniques , Meningitis, Cryptococcal/epidemiology , Molecular Epidemiology , Cryptococcus neoformans/classification , Tertiary Care Centers , Genotype
6.
Rev. Soc. Bras. Med. Trop ; 51(3): 352-356, Apr.-June 2018. tab
Article in English | LILACS | ID: biblio-1041467

ABSTRACT

Abstract INTRODUCTION We describe the clinical and laboratorial features of oral candidiasis in 66 HIV-positive patients. METHODS: Polymerase chain reaction-based techniques were performed for differentiation of Candida spp. isolated from patients at a public teaching hospital in Midwest Brazil. RESULTS: Oral lesions, mainly pseudomembranous, were significantly related to higher levels of immunosuppression. Of 45 Candida isolates, 66.7% were C. albicans. Most of the isolates were susceptible to the antifungal drugs tested. CONCLUSIONS: Oral lesions were associated with higher immunosuppression levels. Lower susceptibility to antifungals by non-albicans isolates supports the importance of surveillance studies using susceptibility tests to aid in the treatment.


Subject(s)
Humans , Male , Female , Adult , Young Adult , Candida/drug effects , Candidiasis, Oral/diagnosis , AIDS-Related Opportunistic Infections/diagnosis , Antifungal Agents/pharmacology , Brazil , Candida/isolation & purification , Candida/classification , Candidiasis, Oral/microbiology , Drug Resistance, Microbial , Microbial Sensitivity Tests , Fluconazole/pharmacology , Amphotericin B/pharmacology , Mycological Typing Techniques , AIDS-Related Opportunistic Infections/microbiology , Itraconazole/pharmacology , Middle Aged
7.
Braz. j. microbiol ; 49(supl.1): 193-198, 2018. tab, graf
Article in English | LILACS | ID: biblio-974340

ABSTRACT

Abstract In this study, phenotypic methods presented >80% agreement with the molecular identification of 59 Candida parapsilosis complex. Growth at 15% NaCl or pH 7.0 significantly reduced cfu-counts of Candida orthopsilosis, suggesting these conditions may support the development of phenotypic methods for the differentiation of the cryptic species of C. parapsilosis complex.


Subject(s)
Humans , Candidiasis/microbiology , Mycological Typing Techniques/methods , Candida parapsilosis/isolation & purification , Phenotype , Polymerase Chain Reaction , Culture Media/metabolism , Candida parapsilosis/classification , Candida parapsilosis/growth & development , Candida parapsilosis/genetics
8.
Mem. Inst. Oswaldo Cruz ; 112(3): 214-219, Mar. 2017. tab, graf
Article in English | LILACS | ID: biblio-1040568

ABSTRACT

Since the description of Candida orthopsilosis and C. metapsilosis in 2005, several methods have been proposed to identify and differentiate these species from C. parapsilosis sensu stricto. Species-specific uniplex polymerase chain reaction (PCR) was performed and compared with sequencing of the D1/D2 region of the LSU 28S rDNA gene, microsatellite typing of C. parapsilosis sensu stricto, and PCR-restriction fragment length polymorphism patterns in the ITS1-5.8S-ITS2 region of the rDNA gene. There was agreement between results of testing of 98 clinical isolates with the four PCR-based methods, with 59 isolates identified as C. parapsilosis sensu stricto, 37 as C. orthopsilosis, and two as C. metapsilosis.


Subject(s)
Humans , Candida/isolation & purification , Mycological Typing Techniques/methods , Polymorphism, Restriction Fragment Length , Candida/classification , Candida/genetics , DNA, Fungal/analysis , Polymerase Chain Reaction , DNA Fingerprinting , Sequence Analysis, DNA , DNA, Ribosomal Spacer/genetics , Genotype
9.
Article in English | LILACS | ID: biblio-842772

ABSTRACT

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


Subject(s)
Humans , Child , AIDS-Related Opportunistic Infections/microbiology , Candida/genetics , Candidiasis, Oral/microbiology , DNA, Fungal/genetics , Candida albicans/genetics , Candida albicans/isolation & purification , Candida/classification , Candida/isolation & purification , Genotype , Mouth Mucosa/microbiology , Mycological Typing Techniques , Phenotype , Polymerase Chain Reaction
11.
Mem. Inst. Oswaldo Cruz ; 111(10): 642-648, Oct. 2016. tab, graf
Article in English | LILACS | ID: lil-796905

ABSTRACT

The propagules of the fungal species Cryptococcus neoformans and C. gattii, whose varieties are distributed world wide, are the primary cause of cryptococcosis, a life threatening disease. The study of environmental and clinical isolates of Cryptococcosis is an important contribution to the epidemiology and ecology of the fungus. The aim of this work was to determine the presence of C. neoformans and C. gattii in the environment in Bogotá, Colombia’s capital city and to establish the relation between clinical and environmental isolates in the period 2012-2015. From a total of 4.116 environmental samples collected between October 2012 - March 2014, 35 were positive for C. neoformans var. grubii. From 55 cryptococcosis cases reported in Bogotá during 2012-2015, 49 isolates were recovered. From those, 94% were identified as C. neoformans var. grubii molecular type VNI; 4% as VNII and 1,2% as C. neoformans var neoformans VNIV. The 84 detected clinical and environmental isolates studied had a similarity between 49-100% according with molecular typing. The correlation between environmental and clinical samples confirms the hypothesis that patients acquire the disease from environmental exposure to the fungal propagules.


Subject(s)
Humans , Male , Female , Cryptococcosis/microbiology , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , Environmental Microbiology , Cities , Colombia , Cryptococcus gattii/genetics , Cryptococcus neoformans/genetics , DNA Fingerprinting , DNA, Fungal , Genotype , Molecular Typing , Mycological Typing Techniques
12.
Braz. j. microbiol ; 47(1): 181-190, Jan.-Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-775120

ABSTRACT

Abstract In the present work we isolated and identified various indigenous Saccharomyces cerevisiae strains and screened them for the selected oenological properties. These S. cerevisiae strains were isolated from berries and spontaneously fermented musts. The grape berries (Sauvignon blanc and Pinot noir) were grown under the integrated and organic mode of farming in the South Moravia (Czech Republic) wine region. Modern genotyping techniques such as PCR-fingerprinting and interdelta PCR typing were employed to differentiate among indigenous S. cerevisiae strains. This combination of the methods provides a rapid and relatively simple approach for identification of yeast of S. cerevisiae at strain level. In total, 120 isolates were identified and grouped by molecular approaches and 45 of the representative strains were tested for selected important oenological properties including ethanol, sulfur dioxide and osmotic stress tolerance, intensity of flocculation and desirable enzymatic activities. Their ability to produce and utilize acetic/malic acid was examined as well; in addition, H2S production as an undesirable property was screened. The oenological characteristics of indigenous isolates were compared to a commercially available S. cerevisiae BS6 strain, which is commonly used as the starter culture. Finally, some indigenous strains coming from organically treated grape berries were chosen for their promising oenological properties and these strains will be used as the starter culture, because application of a selected indigenous S. cerevisiae strain can enhance the regional character of the wines.


Subject(s)
Saccharomyces cerevisiae/classification , Saccharomyces cerevisiae/isolation & purification , Vitis/microbiology , Acetic Acid/metabolism , Bacterial Adhesion , Czech Republic , DNA Fingerprinting , Drug Tolerance , Ethanol/toxicity , Hydrogen Sulfide/metabolism , Molecular Typing , Mycological Typing Techniques , Malates/metabolism , Osmotic Pressure , Polymerase Chain Reaction , Stress, Physiological , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/physiology , Sulfur Dioxide/toxicity
13.
Braz. j. microbiol ; 47(1): 225-230, Jan.-Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-775131

ABSTRACT

Abstract The Van cat is a domestic landrace found in the Van province of eastern Turkey. In this study, we aimed to determine the seasonal carriage of dermatophytes in Van cats without clinical lesions. A total of 264 hair specimens were collected from clinically healthy cats in and around the Van Province. Of these samples, 30.3% were obtained in spring, 30.6% in summer, 16.6% in autumn, and 22.3% in winter; 45.1% of samples were from male cats and the rest from female ones. Of the studied cats, 118 were younger than 1 year, 78 were 1–3 years old, and 68 were older than 3 years. The specimens were subjected to direct microscopic examination with 15% potassium hydroxide and cultured on Sabouraud dextrose agar and dermatophyte test medium supplemented with cycloheximide and chloramphenicol. Dermatophyte identification was carried out based on macroscopic and microscopic colony morphology, urease activities, in vitro hair perforation test, growth at 37 °C, and pigmentation on corn meal agar. Dermatophytes were isolated from 19 (7.1%) of the 264 specimens examined. The most frequently isolated fungi were Trichophyton terrestre (4.1%), followed by Microsporum gypseum (1.1%), M. nanum (1.1%), and T. mentagrophytes (0.7%), and these fungi may represent a health risk for humans in contact with clinically healthy Van cats. M. canis was not isolated from any of the specimens. Our results show no significant (p > 0.05) association between carriage of dermatophytes and the gender of cats. The carriage rate of dermatophytes was high in spring and winter, and the only possible risk factor for infection was age of the animal.


Subject(s)
Animals , Cats , Female , Male , Arthrodermataceae/classification , Arthrodermataceae/isolation & purification , Carrier State/veterinary , Hair/microbiology , Tinea/veterinary , Arthrodermataceae/growth & development , Carrier State/microbiology , Culture Media/chemistry , Microbiological Techniques , Microscopy , Mycological Typing Techniques , Pigments, Biological , Turkey , Tinea/microbiology
14.
Med Mycol ; 54(1): 97-102, 2016. ilus, tab
Article in English | SES-SP, LILACS, SES-SP, SESSP-IALPROD, SES-SP, SESSP-IALACERVO | ID: biblio-1022473

ABSTRACT

Cryptococcal infection is transmitted by the inhalation of Cryptococcus spp. propagules. Information about the Cryptococcus species inhabiting plants might be clinically relevant due to the epidemiological role of these habitats as possible sources of human infection. The aim of this study was to increase the knowledge about the environmental occurrence of cryptococcosis agents. Hollow tree vegetal debris of nine plant species was sampled quarterly over a 12-month period. Melanized colonies were screened for Cryptococcus neoformans and C. gattii by biochemical tests, followed by URA5-RFLP molecular analysis, M13 fingerprinting assays, and mating-typing with the specific a and α primers. The susceptibility to fluconazole of all of the confirmed species colonies was determined using the AFST-EUCAST broth dilution method. We found that the typical Brazilian flora tree Hymenaea courbaril yielded a high cryptococcal burden (median, 10(2) CFU/g) during the summer, autumn and winter seasons. C. neoformans VNI molecular type MAT alpha was identified in all of the samples. The fingerprinting analyses showed great molecular variability with no correlation with the susceptibility profile to fluconazole (MIC range 4 to ≥64 mg/l). To our knowledge, this study is the first describing the association between C. neoformans and Hymenaea courbaril. These observations extend the known geographic distribution of and substantiate a new urban environmental niche for C. neoformans and also emphasize the genetic diversity of the environmental C. neoformans VNI molecular type isolates.


Subject(s)
Seasons , Genetic Variation , Wood/microbiology , Polymorphism, Restriction Fragment Length , Brazil , Colony Count, Microbial , Microbial Sensitivity Tests , Fluconazole/pharmacology , Mycological Typing Techniques , Cryptococcus neoformans , Cryptococcus neoformans/isolation & purification , Cryptococcus neoformans/classification , Cryptococcus neoformans/physiology , Genes, Mating Type, Fungal , Hymenaea/microbiology , Molecular Typing , Genotype , Antifungal Agents/pharmacology
15.
Article in English | WPRIM | ID: wpr-37148

ABSTRACT

Mucormycosis, a fatal opportunistic infection in immunocompromised hosts, is caused by fungi belonging to the order Mucorales. Early diagnosis based on exact identification and multidisciplinary treatments is critical. However, identification of Mucorales fungi is difficult and often delayed, resulting in poor prognosis. This study aimed to compare the results of phenotypic and molecular identification of 12 Mucorales isolates collected from 4-yr-accumulated data. All isolates were identified on the basis of phenotypic characteristics such as growth rate, colony morphology, and reproductive structures. PCR and direct sequencing were performed to target internal transcribed spacer (ITS) and/or D1/D2 regions. Target DNA sequencing identified five Lichtheimia isolates, two Rhizopus microsporus isolates, two Rhizomucor pusillus isolates, one Cunninghamella bertholletiae isolate, one Mucor fragilis isolate, and one Syncephalastrum racemosum isolate. Five of the 12 (41.7%) isolates were incorrectly identified on the basis of phenotypic identification. DNA sequencing showed that of these five isolates, two were Lichtheimia isolates, one was Mucor isolate, one was Rhizomucor isolate, and one was Rhizopus microspores. All the isolates were identified at the species level by ITS and/or D1/D2 analyses. Phenotypic differentiation and identification of Mucorales is difficult because different Mucorales share similar morphology. Our results indicate that the molecular methods employed in this study are valuable for identifying Mucorales.


Subject(s)
Genotype , Humans , Mucorales/classification , Mucormycosis/microbiology , Mycological Typing Techniques , Phenotype
16.
Rev. cuba. med. trop ; 67(3): 0-0, dic. 2015. ilus, tab
Article in Spanish | LILACS, CUMED | ID: lil-777073

ABSTRACT

Introducción: la tiña negra es una micosis superficial causada por el hongo Hortaea werneckii. Se considera una micosis benigna que por lo general es observada en países tropicales. Objetivo: reportar siete casos de tiña negra en niños de dos hospitales de La Habana, Cuba. Métodos: se realizó estudio micológico (examen directo y cultivo) a partir de escamas tomadas mediante raspado de las lesiones a siete niños con diagnóstico clínico presuntivo de tiña negra palmar. Se registraron las características de las lesiones, edad, sexo y factores predisponentes de los pacientes, así como la evolución del cuadro con el tratamiento antifúngico. Resultados: se confirmó la sospecha clínica de tiña negra a través del aislamiento e identificación de Hortae werneckii. Las edades de los pacientes oscilaron entre 3 y 6 años y el 57 por ciento era del sexo femenino. La hiperhidrosis se encontró en el 43 por ciento de los casos. El tratamiento específico con antifúngicos azólicos y terbinafina tópicos fue satisfactorio en 21 días como promedio. Conclusiones: todos los casos con sospecha de tiña negra fueron confirmados de manera oportuna en el laboratorio, lo que permitió descartar enfermedades malignas y aplicar tratamiento específico(AU)


Subject(s)
Humans , Child, Preschool , Child , Tinea/diagnosis , Tinea/drug therapy , Mycological Typing Techniques , Mycoses/diagnosis
17.
Rev. Soc. Bras. Med. Trop ; 48(5): 580-586, Sept.-Oct. 2015. tab, graf
Article in English | LILACS | ID: lil-763328

ABSTRACT

ABSTRACTINTRODUCTION:Cryptococcosis is an invasive disease acquired by inhalation of infectious propagules from the environment. Currently, compulsory notification of the spread of this disease is not required in Colombia. However, reporting of human immunodeficiency virus (HIV)/acquired immune deficiency syndrome cases to the National Surveillance System has suggested that there is a growing population at risk of contracting cryptococcosis. Few studies have described the occurrence of cryptococcosis in Colombia. Therefore, in this study, we examined the pathology of this disease in Atlántico, Colombia and determined the distributions of Cryptococcus neoformans and Cryptococcus gattii in the environment.METHODS:Clinical samples/isolates were gathered from cases of cryptococcosis previously diagnosed at health institutions in Atlántico, and surveys were completed by clinicians. The environmental study considered 32 sampling points and three tree species, i.e., Quickstick ( Gliricidia sepium ), Almond ( Terminalia catappa ), and Pink trumpet ( Tabebuia rosea ). Environmental and clinical samples/isolates were analyzed for phenotypic and genotypic confirmation.RESULTS:From 1997-2014, 41 cases of cryptococcosis were reported. The mean patient age was 40.5 years (range: 18-63 years); 76% were men, and 78% were HIV positive. Isolation was possible in 38 cases ( C. neoformans , molecular type VNI in 37 cases and C. gattii , molecular type VGI in one case). In 2012-2014, 2,068 environmental samples were analyzed with a positivity of 0.4% ( C. neoformans , molecular type VNI) in Almond and Pink trumpet trees.CONCLUSIONS:Cryptococcus neoformans , molecular type VNI had a higher prevalence than C. gattii and was associated with human exposure and the pathogenesis of cryptococcosis in this geographical region.


Subject(s)
Adult , Humans , Male , Middle Aged , Young Adult , Cryptococcosis/epidemiology , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , Environmental Microbiology , Colombia/epidemiology , Cryptococcosis/transmission , Genotype , Molecular Typing , Mycological Typing Techniques , Prevalence
18.
Braz. j. microbiol ; 46(3): 911-920, July-Sept. 2015. tab, ilus
Article in English | LILACS | ID: lil-755798

ABSTRACT

A new inulinase-producing strain was isolated from rhizosphere soils of Jerusalem artichoke collected from Shihezi (Xinjiang, China) using Jerusalem artichoke power (JAP) as sole carbon source. It was identified as an Aspergillus niger strain by analysis of 16S rRNA. To improve inulinase production, this fungus was subjected to mutagenesis induced by 60Co γ-irradiation. A genetically stable mutant (designated E12) was obtained and it showed 2.7-fold higher inulinase activity (128 U/mL) than the parental strain in the supernatant of a submerged culture. Sequential methodology was used to optimize the inulinase production of stain E12. A screening trial was first performed using Plackett-Burman design and variables with statistically significant effects on inulinase bio-production were identified. These significant factors were further optimized by central composite design experiments and response surface methodology. Finally, it was found that the maximum inulinase production (185 U/mL) could be achieved under the optimized conditions namely pH 7.0, yeast extract concentration of 5.0 g/L, JAP concentration of 66.5 g/L, peptone concentration of 29.1 g/L, solution volume of 49.4 mL in 250-mL shake flasks, agitation speed of 180 rpm, and fermentation time of 60 h. The yield of inulinase under optimized culture conditions was approximately 1.4-fold of that obtained by using basal culture medium. These findings are of significance for the potential industrial application of the mutant E12.

.


Subject(s)
Aspergillus niger/enzymology , Aspergillus niger/genetics , Bioreactors/microbiology , Glycoside Hydrolases/metabolism , Helianthus/microbiology , Aspergillus niger/metabolism , China , Culture Media , Ethanol/metabolism , Fermentation/physiology , Inulin/metabolism , Molecular Typing , Mutation , Mycological Typing Techniques , Rhizosphere , /genetics , Soil Microbiology
19.
Braz. j. microbiol ; 46(3): 815-823, July-Sept. 2015. tab, ilus
Article in English | LILACS | ID: lil-755801

ABSTRACT

Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO2 concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking.

.


Subject(s)
Ascomycota/enzymology , Cryptococcus/enzymology , Polygalacturonase/metabolism , Rhodotorula/enzymology , Vitis/microbiology , Wine/microbiology , Argentina , Ascomycota/isolation & purification , Cryptococcus/isolation & purification , Fermentation/physiology , Molecular Sequence Data , Molecular Typing , Mycological Typing Techniques , Polymerase Chain Reaction , Pectins/metabolism , RNA, Ribosomal/genetics , Rhodotorula/isolation & purification
20.
Braz. j. microbiol ; 46(3): 903-910, July-Sept. 2015. tab, ilus
Article in English | LILACS | ID: lil-755814

ABSTRACT

Nineteen fungi and seven yeast strains were isolated from sugarcane bagasse piles from an alcohol plant located at Brazilian Cerrado and identified up to species level on the basis of the gene sequencing of 5.8S-ITS and 26S ribosomal DNA regions. Four species were identified: Kluyveromyces marxianus, Aspergillus niger, Aspergillus sydowii and Aspergillus fumigatus, and the isolates were screened for the production of key enzymes in the saccharification of lignocellulosic material. Among them, three strains were selected as good producers of hemicellulolitic enzymes: A. niger (SBCM3), A. sydowii (SBCM7) and A. fumigatus (SBC4). The best β-xylosidase producer was A. niger SBCM3 strain. This crude enzyme presented optimal activity at pH 3.5 and 55 °C (141 U/g). For β-glucosidase and xylanase the best producer was A. fumigatus SBC4 strain, whose enzymes presented maximum activity at 60 °C and pH 3.5 (54 U/g) and 4.0 (573 U/g), respectively. All these crude enzymes presented stability around pH 3.0–8.0 and up to 60 °C, which can be very useful in industrial processes that work at high temperatures and low pHs. These enzymes also exhibited moderate tolerance to ethanol and the sugars glucose and xylose. These similar characteristics among these fungal crude enzymes suggest that they can be used synergistically in cocktails in future studies of biomass conversion with potential application in several biotechnological sectors.

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Subject(s)
Aspergillus fumigatus/enzymology , Aspergillus niger/enzymology , Cellulose/metabolism , /metabolism , Kluyveromyces/enzymology , Saccharum/microbiology , Xylosidases/metabolism , beta-Glucosidase/metabolism , Aspergillus fumigatus/isolation & purification , Aspergillus fumigatus/metabolism , Aspergillus niger/isolation & purification , Aspergillus niger/metabolism , Base Sequence , Biomass , Brazil , DNA, Fungal/genetics , DNA, Intergenic/genetics , Fermentation , Kluyveromyces/isolation & purification , Kluyveromyces/metabolism , Lignin/metabolism , Molecular Typing , Mycological Typing Techniques , RNA, Ribosomal/genetics , Sequence Analysis, DNA
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