ABSTRACT
ABSTRACT Background: This study aims to explore the epidemiology, clinical profile and strain characteristics of cryptococcosis from 2013 to 2017 in a major teaching hospital in China. Methods: Trends in antifungal drug susceptibility of 217 consecutive non-repetitive cryptococcal isolates collected from patients of an university hospital in China were analyzed between 2013 and 2017. Of those, 98 isolates were conserved for identification by internal transcribed spacer (ITS) sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) system. Multilocus sequence typing (MLST) was used to designate molecular types. Clinical characteristics of the 98 patients with cryptococcosis during the period of 2013-2017 were retrospectively evaluated. Results: There was a trend for gradual increase in the MIC range of fluconazole was from 2013 to 2017. The conserved 98 clinical cryptococcal isolates included 97 C. neoformans and one C. gattii, and 90 (91.8%) isolates belonged to ST5 genotype VNI. Out of the 98 patients with cryptococcosis, 28 (28.6%) were HIV-infected and 32 (32.7%) had no underlying diseases. HIV-infected patients had higher mortality than HIV-uninfected patients (28.6% vs 14.3%, p = 0.147). Conclusions: Most of the patients with cryptococcosis were not HIV-infected in this study, while patients with HIV had a higher mortality. Reduced susceptibility to fluconazole was observed among C. neoformans isolates, most of them belonged to ST5 genotype VNI having an impact on the effective dose of fluconazole.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Cryptococcosis/microbiology , Cryptococcosis/epidemiology , Hospitals, University/statistics & numerical data , Time Factors , Microbial Sensitivity Tests , China/epidemiology , Cross-Sectional Studies , Retrospective Studies , Statistics, Nonparametric , Cryptococcosis/drug therapy , Cryptococcus neoformans/isolation & purification , Cryptococcus neoformans/drug effects , Cryptococcus neoformans/genetics , Cryptococcus gattii/isolation & purification , Cryptococcus gattii/drug effects , Cryptococcus gattii/genetics , Multilocus Sequence Typing , Genotype , Antifungal Agents/therapeutic useABSTRACT
Abstract INTRODUCTION Cryptococcosis is the second most frequent cause of opportunistic infections in human immunodeficiency virus (HIV) patients in Colombia. We aimed to determine the prevalence of cryptococcosis in the Colombian department of Atlántico. METHODS An active search for cryptococcosis cases was conducted between 2015 and 2017 in health institutions by distributing surveys to clinicians and characterizing samples phenotypically and genotypically. RESULTS Thirty-eight cryptococcosis cases were identified (81.6% men, 76.3% HIV patients). The calculated annual prevalence was 5.08/1 million inhabitants. Cryptococcus neoformans var. grubii VNI was isolated in 34 cases. CONCLUSIONS These results provide the basis for passive surveillance of cryptococcosis.
Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Young Adult , Cryptococcosis/epidemiology , Phenotype , Prevalence , Colombia/epidemiology , Cryptococcosis/microbiology , Cryptococcus neoformans/isolation & purification , Cryptococcus gattii/isolation & purification , Molecular Typing , Genotype , Middle AgedABSTRACT
La criptococosis es una micosis grave de distribución universal, que afecta principalmente a huéspedes inmunocomprometidos. Es una de las principales causas de morbilidad y mortalidad en los pacientes infectados con el virus de la inmunodeficiencia humana (HIV). Provoca al menos 620 000 muertes al año, representando entre el 13% al 44% de la mortalidad en pacientes HIV positivos según datos de cohortes correspondientes a países en desarrollo. (1, 2) La letalidad de la criptococosis meníngea en estudios de Argentina y Brasil muestra valores que van desde el 26% hasta el 63%. El complejo Cryptococcus neoformans/ Cryptococcus gattii, es el responsable de esta enfermedad. Existen alrededor de 70 especies pero solo dos de ellas son patógenas para el hombre: C. neoformans y C. gattii. Se reconocen 8 genotipos de este complejo, C. neoformans: VNI y VNII (C. neoformans var. grubii), VNIII (C. neoformans híbrido intervariedad AD), VNIV (C. neoformans var. neoformans) y C. gattii: genotipos VGI, VGII, VGIII y VGIV. Se han descripto híbridos interespecie VNIV/VGI, VNI/VGI, VNI/VGII. Se estudiaron 207 aislamientos de Cryptococcus, elegidos aleatoriamente, de un total de 2593 pacientes con diagnóstico de criptococosis diseminada. A los mismos se les realizó la genotipificación mediante una PCR-RFLP del gen URA5, y posterior digestión enzimática con enzimas Sau96I y HhaI. De las 207 cepas estudiadas, 174 fueron VNI (84,05%), 14 VNII (6,76%), 10 VNIII (4,83%), 2 VNIV (0,97%), 3 VGI (1,45%), 3 VGII de (1,45%) y 1 VGIII (0,49%).
Cryptococcosis is a severe worldwide mycosis, which mainly affects immunocompromised hosts and is a major cause of morbidity and mortality in HIV-infected patients. It causes 620,000 annual deaths, accounting for 13-44 % of mortality in HIV-positive individuals in developing countries. Mortality rates of meningeal cryptococcosis in studies from Argentina and Brazil go from 26 to 63 %. Cryptococcus neoformans/Cryptococcus gattii is the species complex responsible for this disease. There are about 70 species, however, only two are human pathogens: C. neoformans and C. gattii. C. neoformans genotypes are VNI and VNII (C. neoformans var. grubii), VNIII (C. neoformans intervariety hybrid AD), VNIV (C. neoformans var. neoformans). C. gattii genotypes are VGI, VGII, VGIII and VGIV. Interspecies hybrids were described: VNIV/VGI, VNI/VGI, VNI/ VGII. A total of 207 Cryptococcus isolates were randomly selected from 2593 patients with diagnosis of disseminated cryptococcosis. Genotyping was performed by PCRRFLP of UR A5 gene with restriction enzyme digestion using Sau96I and HhaI enzymes. Among the 207 studied isolates, 174 resulted VNI (84.05%), 14 VNII (6.76%), 10 VNIII (4.83%), 2 VNIV (0.97%), 3 VGI (1.45%), 3 VGII (1.45%) and 1 VGIII (0.49%).
Subject(s)
Humans , Cross-Sectional Studies/statistics & numerical data , Morbidity , HIV/isolation & purification , Meningitis, Cryptococcal/epidemiology , Cryptococcus neoformans/isolation & purification , Cryptococcus gattii/isolation & purification , GenotypeABSTRACT
Abstract INTRODUCTION: This study aimed to describe cryptococcal meningitis (CM) cases and the associated demographic, clinical, and microbiological data obtained from cities in the State of Mato Grosso do Sul in the Midwestern region of Brazil. METHODS: The data from 129 patients with laboratory-confirmed CM admitted from 1997 to 2014 were retrospectively reviewed. The molecular types of Cryptococcus neoformans and Cryptococcus gattii isolated from cerebrospinal fluid were analyzed to determine their geographic distribution. RESULTS: The patients had a mean age of 37 years and consisted mostly of men (76.7%). Most of the Cryptococcus isolates were obtained from patients infected with human immunodeficiency virus (HIV) and included 105 (87.5%) and 5 (55.6%) isolates of C. neoformans and C. gattii complexes, respectively. A restriction fragment length polymorphism (RFLP) analysis of URA5 revealed that most of the isolates were C. neoformans molecular type VNI (89.1%), whereas the molecular types VGII (7%) and VNII (3.9%) were observed less frequently. Notably, 65% of the cases with a time from symptom onset to laboratory diagnosis of more than 60 days resulted in fatalities, and sequelae were observed among the patients who survived. CONCLUSIONS: The present study documents the occurrence of neurocryptococcosis, which is mainly caused by C. neoformans VNI, in Mato Grosso do Sul, Brazil, with probable autochthonous cases in the Brazilian Pantanal, the world's largest tropical wetland, and a biome where cryptococcosis has not yet been explored.
Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Young Adult , DNA, Fungal/analysis , Meningitis, Cryptococcal/epidemiology , Cryptococcus neoformans/isolation & purification , Cryptococcus gattii/isolation & purification , Rural Population , Socioeconomic Factors , Urban Population , Brazil/epidemiology , DNA, Fungal/cerebrospinal fluid , Polymerase Chain Reaction , Retrospective Studies , Meningitis, Cryptococcal/cerebrospinal fluid , Meningitis, Cryptococcal/microbiology , Cryptococcus neoformans/genetics , Cryptococcus gattii/genetics , Genotype , Middle AgedABSTRACT
ABSTRACT Cryptococcosis is a major invasive fungal disease related worldwide with the AIDS population. New reports of HIV/AIDS cases to the national public health surveillance system (SIVIGILA) in Colombia have shown that there is a growing community at risk of contracting cryptococcosis throughout the country who do not have access to ART. Even though the most prevalent species Cryptococcus neoformans is mainly associated with the HIV population, we report a fatal case of cryptococcosis in an AIDS patient in Barranquilla, associated with Cryptococcus gattii VGI, isolated from blood culture.
Subject(s)
Humans , Female , Middle Aged , AIDS-Related Opportunistic Infections/microbiology , Cryptococcosis/diagnosis , Cryptococcus gattii/genetics , AIDS-Related Opportunistic Infections/diagnosis , Colombia , Cryptococcus gattii/isolation & purification , Fatal Outcome , GenotypeABSTRACT
Abstract Cryptococcosis is a fungal infection of opportunistic behavior that is unusual in immunocompetent patients. We report a rare case of disseminated cryptococcosis with cutaneous involvement in an immunocompetent individual. During hospitalization, Cryptococcus gattii was isolated from skin lesions, lung and spinal fluid. The diagnosis of disseminated cryptococcosis was confirmed and treatment was established. The patient showed improvement. Due to the probable clinical severity of the disease and the possibility that skin lesions may be the first manifestation of this illness, prompt diagnosis must be established and treatment provided.
Subject(s)
Humans , Male , Adult , Cryptococcosis/immunology , Cryptococcosis/pathology , Dermatomycoses/immunology , Dermatomycoses/pathology , Cryptococcus gattii/isolation & purification , Immunocompetence , Skin/microbiology , Skin/pathology , Treatment Outcome , Cryptococcosis/drug therapy , Dermatomycoses/drug therapy , Lymphocytosis/complications , Lung/microbiology , Antifungal Agents/therapeutic useSubject(s)
Humans , Male , Female , Cryptococcosis/microbiology , Cryptococcus neoformans/isolation & purification , Environmental Microbiology , Cryptococcus gattii/isolation & purification , DNA, Fungal , DNA Fingerprinting , Cities , Mycological Typing Techniques , Colombia , Cryptococcus neoformans/genetics , Cryptococcus gattii/genetics , Molecular Typing , GenotypeABSTRACT
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 TechniquesABSTRACT
Introducción: la criptocococis es una micosis sistémica causada por C. neoformans y C. gattii, es frecuente y oportunista en inmunocomprometidos y patógeno primario en personas inmunocompetentes. C. neoformans tiene una distribución mundial y se ha aislado desde las excretas de palomas. C. gattii se considera restringida a regiones con clima tropical, subtropical, y templadas, se encuentra asociada frecuentemente a detritos de especies de Eucalyptus sp. La virulencia de estas levaduras le permite desarrollar patogénesis en mamíferos y supervivencia en el ambiente. Objetivo: Identificar y determinar la actividad de proteinasas y fosfolipasas, de C. neoformans y C. gattii aisladas desde las oquedades de árboles en lugares con alta afluencia de público. Materiales y Métodos: Se tomaron 200 muestras de hisopado desde distintas especies de árboles desde sectores de la región de OHiggins y el Maule. Se siembran en ASG, se aíslan y mantienen en ASD. Identificación con tinta china, Urea de Christensen, crecimiento a 37°C, asimilación y fermentación de azucares, y siembra en medio CGB. Se mide índice de actividad enzimática Prz de proteinasas y fofolipasas. Resultados y Conclusiones: Se obtuvieron 109 cepas de C. neoformans aisladas desde las oquedades de diferentes especies arbóreas y 3 cepas presuntivas de C. gattii desde Eucalyptus sp. y Prunus cerasifera artropurpurea. El 88,1 por ciento de las cepas C. neoformans y 100 por ciento de C. gattii, presentaron alta actividad proteolítica, El 49,5 por ciento de las cepas de C. neoformans y 33,3 por ciento de C. gattii mostraron alta actividad de fosfolipasas.
Introduction: criptocococis is a systemic mycosis caused by C. neoformans and C. gattii, frequent and opportunistic in immunocompromised and primary pathogen in immunocompetent persons. C. neoformans has a worldwide distribution and has been isolated from the excreta of pigeons. C. gattii is considered restricted to regions with tropical, subtropical, and temperate, is often associated with species of Eucalyptus sp. The virulence of these yeasts develop pathogenesis allows survival in mammals and the environment. Objective: To identify and determine the activity of proteinases and phospholipases of C. neoformans and C. gattii isolated from the hollows of trees in places with high turnout. Materials and Methods: 200 swab samples were taken from different species of trees from areas of the region of OHiggins and Maule. Planted in ASG, they are isolated and kept in ASD. Identification with ink, Urea Christensen, growth at 37 ° C, assimilation and fermentation of sugars, and planting medium CGB. Prz index proteinase enzyme activity is measured and phospholipases. Results and Conclusions: We manage to get 109 strains of C. neoformans isolated from the hollows of different tree species and 3 presumptive strains of C. gattii from Eucalyptus sp. and Prunus cerasifera artropurpurea. 88.1 percent of the strains C. neoformans and C. gattii 100 percent , they showed high proteolytic activity, 49.5 percent of the strains of C. neoformans and C. gattii 33.3 percent showed high activity phospholipases.
Subject(s)
Humans , Cryptococcus gattii/isolation & purification , Cryptococcus gattii/enzymology , Cryptococcus gattii/pathogenicity , Cryptococcus neoformans/isolation & purification , Cryptococcus neoformans/enzymology , Cryptococcus neoformans/pathogenicity , Peptide Hydrolases , Phospholipases , Trees/microbiology , Chile , Cryptococcosis/etiology , Eucalyptus/microbiology , Lung Diseases, Fungal , Prunus/microbiologyABSTRACT
ABSTRACT The diagnosis of cryptococcosis is usually performed based on cultures of tissue or body fluids and isolation of the fungus, but this method may require several days. Direct microscopic examination, although rapid, is relatively insensitive. Biochemical and immunodiagnostic rapid tests are also used. However, all of these methods have limitations that may hinder final diagnosis. The increasing incidence of fungal infections has focused attention on tools for rapid and accurate diagnosis using molecular biological techniques. Currently, PCR-based methods, particularly nested, multiplex and real-time PCR, provide both high sensitivity and specificity. In the present study, we evaluated a nested PCR targeting the gene encoding the ITS-1 and ITS-2 regions of rDNA in samples from a cohort of patients diagnosed with cryptococcosis. The results showed that in our hands, this Cryptococcus nested PCR assay has 100% specificity and 100% sensitivity and was able to detect until 2 femtograms of Cryptococcus DNA.
Subject(s)
Humans , Cryptococcosis/diagnosis , Cryptococcus gattii/genetics , Cryptococcus neoformans/genetics , DNA, Fungal/analysis , Cryptococcosis/microbiology , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , DNA, Ribosomal Spacer , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Sequence Analysis, DNAABSTRACT
In this study, we present antifungal susceptibility data of clinical and environmental isolates of Central Indian Cryptococcus neoformans (Serotype A, n = 8 and n = 50 respectively) and Cryptococcus gattii (Serotype B, n = 01 and n = 04 respectively). Susceptibilities to fluconazole, itraconazole and ketoconazole were determined by using NCCLS broth micro-dilution methodology. The total number of resistant strains for fluconazole in case of C. neoformans and C. gattii showed a significant difference by using chi-square test (p < 0.05*), while considering fisher's exact p value was nonsignificant (p > 0.05). However, the total number of resistant strains for itraconazole and ketoconazole was not found statistically significant. A comparison of geometric means of clinical and environmental strains of C. gattii and C. neoformans was not found statistically significant using student ‘t’ test (p value > 0.05 NS). Though less, the antifungal data obtained in this study suggests that primary resistance among environmental and clinical isolates of C. neoformans and C. gattii against tested antifungal was present and C. gattii comparatively was less susceptible than C. neoformans var. grubii isolates to fluconazole than to itraconazole and ketoconazole. A continuous surveillance of antifungal susceptibility of clinical and environmental isolates of C. neoformans and C. gattii is desirable to monitor the emergence of any resistant strains for better management of cryptococcosis patients.
Subject(s)
Humans , Antifungal Agents/pharmacology , Cryptococcosis/microbiology , Cryptococcus gattii/drug effects , Cryptococcus neoformans/drug effects , Environmental Microbiology , Cryptococcosis/epidemiology , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , Drug Resistance, Fungal , Fluconazole/pharmacology , India/epidemiology , Itraconazole/pharmacology , Ketoconazole/pharmacology , Microbial Sensitivity TestsABSTRACT
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 , PrevalenceABSTRACT
Aim: This study evaluated the use of polymerase chain reaction for cryptococcal meningitis diagnosis in clinical samples. Materials and methods: The sensitivity and specificity of the methodology were evaluated using eight Cryptococcus neoformans/C. gattii species complex reference strains and 165 cere- brospinal fluid samples from patients with neurological diseases divided into two groups: 96 patients with cryptococcal meningitis and AIDS; and 69 patients with other neurological opportunistic diseases (CRL/AIDS). Two primer sets were tested (CN4-CN5 and the multiplex CNa70S-CNa70A/CNb49S-CNb-49A that amplify a specific product for C. neoformans and another for C. gattii). Results: CN4-CN5 primer set was positive in all Cryptococcus standard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattii was observed in the clinical samples of either group. The 695 bp products of C. neoformans were observed only in 64.6% of the cryptococcal meningitis and AIDS group. This primer set was negative for two standard strains. The specificity based on the negative samples from the CTL/AIDS group was 98.5% in both primer sets. Conclusions: These data suggest that the CN4/CN5 primer set was highly sensitive for the identification of C. neoformans/C. gattii species complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis. .
Subject(s)
Humans , Cryptococcus gattii/genetics , Cryptococcus neoformans/genetics , DNA, Fungal/cerebrospinal fluid , Meningitis, Cryptococcal/diagnosis , AIDS-Related Opportunistic Infections/cerebrospinal fluid , AIDS-Related Opportunistic Infections/diagnosis , AIDS-Related Opportunistic Infections/microbiology , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , DNA Primers/genetics , Genotype , Meningitis, Cryptococcal/cerebrospinal fluid , Meningitis, Cryptococcal/microbiology , Polymerase Chain Reaction , Sensitivity and SpecificityABSTRACT
Cryptococcosis is an infection that affects humans and animals, the etiology is attributed to Cryptococcus neoformans variety neoformans, C. neoformans var. grubii and Cryptococcus gattii. The infection is common in dogs and cats, causing respiratory, neurological, cutaneous and ocular infections. Aiming to better understand the epidemiology of cryptococcosis in animals in the region, this paper describe the occurrence and characterization of the Cryptococcus species involved in this illness in pet animals at Mato Grosso State, Brazil. Clinical samples of four cases, two in cats and two dogs, were submitted for pathological, microbiological and molecular analysis. Microscopically, in three cases, tissue sections stained with hematoxylin and eosin had absence to severe granulomatous reaction composed by histiocytes, multinucleated cells and lymphocytes infiltration. In one case, citological imprint analysis showed similar inflammatory mainly mononuclear and lymphocyte cells infiltration. All cases had variable amounts of intracellular and extracellular fungal structures compatible with Cryptococcus sp. on Periodic Acid-Schiff (PAS) stain. All clinical samples were positive for culture on Sabouraud Dextrose Agar (SDA) and morphologically classified as Cryptococcus sp. The isolates were PCR positive for C. gatti, being confirmed by sequencing technique. The findings characterize the molecular species involved in animal infections in the region, and may contribute to future studies of the epidemiology of C. gattii.
A criptococose é uma infecção que afeta humanos e animais sendo a etiologia atribuída às espécies Cryptococcus neoformans variedade neoformans, C. neoformans var. grubii e C. gattii. A doença é comum em cães e gatos, causando infecções respiratórias, neurológicas, cutâneas e oculares. Com o objetivo de entender melhor a epidemiologia da criptococose em animais, este trabalho descreve a ocorrência e a caracterização de espécies de Cryptococcus em pequenos animais doentes no Estado de Mato Grosso, Brasil. Amostras clínicas de quatro casos, dois em felinos e dois em caninos, foram submetidas à análise patológica, microbiológica e molecular. Microscopicamente, em três casos, nos cortes de tecidos corados pela hematoxilina notou-se desde ausência até acentuada inflamação granulomatosa composta por histiócitos, células multinucleadas e infiltração linfocitária. Em um caso, na análise citológica de "imprint" observou-se infiltrado inflamatório similar composto principalmente por células mononucleares e linfócitos. Em todos os casos havia variável quantidade de estruturas fúngicas intra e extracelulares compatíveis com Cryptococcus spp pela coloração do ácido períodico de Schiff (PAS). Todas as amostras foram positivas para a cultura em Sabouraud Dextrose Agar (SDA), e morfologicamente classificadas como Cryptococcus sp. Os isolados foram positivos no PCR para C. gattii, sendo confirmados pelo seqüenciamento. Os resultados caracterizaram a espécie envolvida na região, e contribuem para futuros estudos sobre a epidemiologia de C. gattii.
Subject(s)
Animals , Cats , Dogs , Cats , Cryptococcosis/veterinary , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , Dogs , Epidemiologic StudiesABSTRACT
Cryptococcosis is a fungal infection caused by yeast species of Cryptococcus genus, particularly Cryptococcus neoformans/Cryptococcus gattii species complex. The knowledge of the cryptococcosis casuistic in northeastern Argentina is scarce and there is no information about the molecular types circulating in this area. The aim of this study was to genotyping C. neoformans/C. gattii complex clinical isolates obtained at Hospital "Dr. Julio C. Perrando", Resistencia city (Chaco, Argentina), in order to determine species, variety and molecular type. During two years and one month 26 clinical isolates were studied. Using conventional and molecular methods one isolate was identified as C. gattii VGI type, and 25 isolates as C. neoformans var. grubii; 23 of these belonged to VNI type and two belonged to VNII type. This data is a contribution to the knowledge of cryptococcosis epidemiology in Argentina and the first report about C. neoformans/ C. gattii complex molecular types from clinical isolates in northeastern Argentina.
Subject(s)
Cryptococcus gattii/genetics , Cryptococcus neoformans/genetics , Adult , Young Adult , Argentina , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , Female , Genotype , Hospitals , Humans , Male , Middle AgedABSTRACT
In this study, we evaluated several techniques for the detection of the yeast form of Cryptococcus in decaying wood and measured the viability of these fungi in environmental samples stored in the laboratory. Samples were collected from a tree known to be positive for Cryptococcus and were each inoculated on 10 Niger seed agar (NSA) plates. The conventional technique (CT) yielded a greater number of positive samples and indicated a higher fungal density [in colony forming units per gram of wood (CFU.g-1)] compared to the humid swab technique (ST). However, the difference in positive and false negative results between the CT-ST was not significant. The threshold of detection for the CT was 0.05.10³ CFU.g-1, while the threshold for the ST was greater than 0.1.10³ CFU-1. No colonies were recovered using the dry swab technique. We also determined the viability of Cryptococcus in wood samples stored for 45 days at 25ºC using the CT and ST and found that samples not only continued to yield a positive response, but also exhibited an increase in CFU.g-1, suggesting that Cryptococcus is able to grow in stored environmental samples. The ST.1, in which samples collected with swabs were immediately plated on NSA medium, was more efficient and less laborious than either the CT or ST and required approximately 10 min to perform; however, additional studies are needed to validate this technique.
Subject(s)
Cryptococcus gattii/growth & development , Cryptococcus neoformans/growth & development , Microbial Viability , Specimen Handling/methods , Wood/microbiology , Colony Count, Microbial , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/isolation & purification , Environmental MicrobiologyABSTRACT
Introduction: A survey on cryptococcosis is being conducted regularly in Colombia since 1997. We present hereby the results corresponding to patients diagnosed from 2006 to 2010. Objective: To analyze the data obtained during this period. Materials and methods: Retrospective analysis of the corresponding surveys. Results: A total of 526 surveys originating from 72% of the Colombian political divisions were received during the 5-year period. Most patients (76.6%) were males and 74.9% were 21-50 years old. The most prevalent risk factor was HIV infection (83.5%) with cryptococcosis defining AIDS in 23% of the cases. In the general population the estimated mean annual incidence rate for cryptococcosis was 2.4 x 106 inhabitants while in AIDS patients this rate rose to 3.3 x 103. In 474 surveys stating clinical features, most frequent complaints were headache 84.5%, fever 63.4%, nausea and vomiting 57.5%, mental alterations 46.3%, meningeal signs 33.0%, cough 26.4% and visual alterations 24.5%. Neurocryptococcosis was recorded in 81.8% of the cases. Laboratory diagnosis was based on direct examination, culture and latex in 29.3% cases. From 413 Cryptococcus isolates analyzed, 95.6% were identified as C. neoformans var. grubii, 1% C. neoformans var. neoformans, and 3.4% C. gattii. Treatment was reported for 71.6% of the cases with amphotericin B alone or in combination with fluconazole prescribed in 28%. Conclusions: Surveys done through passive surveillance continue to be sentinel markers for HIV infection and represent a systematic approach to the study of opportunistic problems regularly afflicting AIDS patients since cryptococcosis requires no compulsory notification in Colombia.
Introducción. Desde 1997 se viene realizando un programa nacional de vigilancia sobre la criptococosis en Colombia. Se presentan los resultados correspondientes a los pacientes diagnosticados entre el 2006 y el 2010. Objetivo. Analizar los datos obtenidos durante este periodo. Materiales y métodos. Análisis retrospectivo de las encuestas. Resultados. Durante los cinco años mencionados se recibieron 526 encuestas representativas del 72 % de la división política colombiana. La mayoría de pacientes (76,6 %) eran hombres y 74,9 % estaban entre los 21 y los 50 años. El factor de riesgo prevalente fue la infección por VIH (83,5 %), y la criptococosis definió el sida en 23 % de los casos. La incidencia anual promedio en la población general fue de 2,4 por un millón de habitantes mientras que, en pacientes con sida, aumentó a 3,3 por 1.000. En 474 encuestas se informaron manifestaciones clínicas; las más frecuentes fueron: cefalea (84,5 %), fiebre (63,4 %), náuseas y vómito (57,5 %), alteraciones mentales (46,3 %), signos meníngeos (33 %), tos (26,4 %) y alteraciones visuales (24,5 %). La neurocriptococosis se reportó en 81,8 % de los casos. El diagnóstico se hizo por examen directo, cultivo y antigenemia en 29,3 % de los casos. De 413 aislamientos recuperados, 95,6 % fueron C. neoformans var. grubii, 1 % C. neoformans var. neoformans, y 3,4 % C. gattii. En 71,6 % de los casos para el tratamiento se administró anfotericina B y en 28 % se combinó con fluconazol. Conclusiones. La vigilancia pasiva continúa siendo un marcador centinela para la infección por VIH, y constituye una aproximación sistemática al estudio de infecciones oportunistas en pacientes con sida, debido a que la criptococosis no es de notificación obligatoria en Colombia.
Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Cryptococcosis/epidemiology , AIDS-Related Opportunistic Infections/epidemiology , Amphotericin B/administration & dosage , Amphotericin B/therapeutic use , Antifungal Agents/therapeutic use , Antigens, Fungal/blood , Colombia/epidemiology , Cryptococcosis/diagnosis , Cryptococcosis/drug therapy , Cryptococcosis/microbiology , Cryptococcus gattii/immunology , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/immunology , Cryptococcus neoformans/isolation & purification , Drug Resistance, Multiple, Fungal , Fluconazole/administration & dosage , Fluconazole/therapeutic use , Health Surveys , Incidence , Population Surveillance , Retrospective Studies , Symptom AssessmentABSTRACT
Cryptococcus spp. are common causes of mycoses in immunocompromised patients. To determine the drug susceptibilities of clinical Cryptococcus spp. isolates, the characteristics of 61 clinical Cryptococcus spp. complex isolates and their antifungal susceptibilities were investigated, including 52 C. neoformans and 9 C. gattii isolates collected at Shanghai between 1993 and 2009. Antifungal susceptibility of clinical isolates to amphotericin B, fluconazole, itraconazole, and flucytosine were determined by the microdilution method M27-A2 and the ATB FUNGUS 3 kit. The 90% minimum inhibitory concentration (MIC90) and susceptibility ranges were as follows: 1 (0.0625-1) µg/mL for amphotericin B, 4 (0.125-16) µg/ mL for fluconazole, 0.25 (0.0313-4) µg/mL for itraconazole, and 4 (0.125-8) µg/mL for flucytosine. Fluconazole, itraconazole, and flucytosine have excellent in vitro activity against all tested clinical Cryptococcus spp., and we also found a high rate of tolerance to amphotericin B (MICs ranging from 0.55-1 µg/mL). Furthermore, C. neoformans isolates from acquired immune deficiency syndrome (AIDS) patients were less susceptible to fluconazole and flucytosine than those from non-AIDS patients. These data suggest that use of amphotericin B may lead to tolerance or resistance of the pathogen over time. There were also no significant associations between species, genotypes, and in vitro susceptibilities of these clinical isolates.
Subject(s)
Humans , AIDS-Related Opportunistic Infections/microbiology , Antifungal Agents/pharmacology , Cryptococcosis/microbiology , Cryptococcus gattii/drug effects , Cryptococcus neoformans/drug effects , China , Cryptococcus gattii/genetics , Cryptococcus gattii/isolation & purification , Cryptococcus neoformans/genetics , Cryptococcus neoformans/isolation & purification , Drug Resistance, Fungal , Genotype , Microbial Sensitivity TestsABSTRACT
Throughout Brazil, Cryptococcus neoformans is the cause of cryptococcosis, whereas Cryptococcus gattii is endemic to the northern and northeastern states. In this study, the molecular types of 63 cryptococcal isolates recovered from the cerebrospinal fluid of meningitis patients diagnosed between 2008-2010 in Teresina, Piauí, Brazil, were analysed. Out of the 63 patients, 37 (58.7 percent) were human immunodeficiency virus (HIV)-positive and 26 (41.3 percent) were HIV-negative. URA5-restriction fragment length polymorphism analysis identified 37/63 (58.7 percent) isolates as the C. neoformans VNI genotype, predominantly in HIV-positive patients (32/37, 86.5 percent), and 24/63 (38.1 percent) as the C. gattii VGII genotype, mostly in HIV-negative patients (21/26, 80.8 percent). The occurrence of C. gattii VGII in six apparently healthy children and in seven adolescents/young adults in this region reaffirms the endemic occurrence of C. gattii VGII-induced primary cryptococcosis and early cryptococcal infection. Lethality occurred in 18/37 (48.6 percent) of the HIV-positive subjects and in 13/26 (50 percent) of the HIV-negative patients. Our results provide new information on the molecular epidemiology of C. neoformans and C. gattii in Brazilian endemic areas.