Clinical and microbiological characteristics of cryptococcosis at an university hospital in China from 2013 to 2017

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.

Synergistic effect of ibuprofen with itraconazole and fluconazole against Cryptococcus neoformans

The present study investigated the association of the non-steroidal anti-inflammatory drug ibuprofen with itraconazole, fluconazole and amphotericin B against Cryptococcus neoformans isolates. The minimal inhibitory concentration (MIC) was found according to M27-A3 protocol and in vitro interactions were evaluated using checkerboard microdilution method. Synergism was demonstrated between azoles and ibuprofen for most isolates. However, no synergistic effects were seen when amphotericin B was combined with ibuprofen. Therefore, our results suggest that ibuprofen presents clinical potential when combined with azole drugs in the treatment of cryptococcosis.

Deep cutaneous ulcers and sinus formation in an immunocompetent adult

Abstract This report describes a case of unusual deep skin ulcers with tortuous sinus tract formation in an immunocompetent woman. She was initially diagnosed with a Staphylococcus aureus skin infection and histopathologically diagnosed with pyoderma gangrenosum. However, culture from the deep end of ribbon gauze inserted into the subcutaneous sinus tract revealed shiny, light-yellow mucoid colonies, which were identified as Cryptococcus neoformans var. grubii. She was treated with fluconazole for nine months and completely healed. Cryptococcosis is an opportunistic infection caused by variants of C. neoformans species. Cutaneous manifestations of cryptococcosis are quite divergent, rarely occurring as deep skin ulcers with sinus formation.

Epidemiological and TNFα polymorphism evaluation in patients with cryptococcal meningitis treated at a referral hospital in North Brazil

Abstract INTRODUCTION: The present study evaluated the epidemiology of cryptococcal meningitis and TNFα gene polymorphisms in patients at a reference hospital in northern Brazil. METHODS: Samples from 25 patients infected with Cryptococcus spp. were collected to confirm the infection and to analyze the TNFα gene polymorphisms. RESULTS: Cryptococcus neoformans was detected as the predominant etiological agent (100%) in HIV-positive patients. No genetic polymorphic changes were found. CONCLUSIONS: No correlation was observed between the analyzed TNFα polymorphisms and cryptococcal meningitis.

Prevalence of cryptococcosis in Atlántico, department of Colombia assessed with an active epidemiological search

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.

Epidemiología molecular de la criptococosis en un Hospital de enfermedades infecciosas de la Ciudad de Buenos Aires: análisis de 25 años / Molecular epidemiology of cryptococcosis in an infectious diseases hospital from the city of Buenos Aires: A 25 years experience

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%).

Cryptococcoma mimicking a brain tumor in an immunocompetent patient: case report of an extremely rare presentation

ABSTRACT CONTEXT: Central nervous system (CNS) infectious diseases have high prevalence in developing countries and their proper diagnosis and treatment are very important for public health planning. Cryptococcus neoformans is a fungus that may cause several CNS manifestations, especially in immunocompromised patients. Cryptococcal meningitis is the most common type of involvement. Mass-effect lesions are uncommon: they are described as cryptococcomas and their prevalence is even lower among immunocompetent patients. The aim here was to report an extremely rare case of cryptococcoma causing a mass effect and mimicking a brain tumor in an immunocompetent patient. The literature on CNS cryptococcal infections was reviewed with emphasis on cryptococcomas. Clinical, surgical and radiological data on a female patient with this rare presentation of cryptococcoma mimicking a brain tumor are described. Case Report: A 54-year-old female patient presented to the emergency department with a rapid-onset progressive history of confusion and completely dependency for basic activities. Neuroimaging showed a left occipital lesion and neurosurgical treatment was proposed. From histopathological evaluation, a diagnosis of cryptococcoma was established. She received clinical support with antifungals, but despite optimal clinical treatment, her condition evolved to death. CONCLUSIONS: Cryptococcal infections have several forms of presentation and, in immunocompetent patients, their manifestation may be even more different. Cryptococcoma is an extremely rare presentation in which proper surgical and clinical treatment should be instituted as quickly as possible, but even so, there is a high mortality rate.

Cryptococcal meningitis epidemiology: 17 years of experience in a State of the Brazilian Pantanal

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.

Criptococcosis en el Hospital Carlos Van Buren de Valparaíso: una serie de casos / Cryptococcosis in Hospital Carlos Van Buren, Valparaiso: a clinical serie

Resumen Introducción: La criptococcosis es una infección micótica oportunista grave, Cryptococcus neoformans es la principal especie de importancia médica, pudiendo manifestarse como meningitis, neumonía o criptococcemia. Objetivo: Caracterizar a los pacientes con infección por Cryptococcus sp. entre el 01/01/13 y 30/06/16, en el HCVB. Materiales y Métodos: Se identificaron los cultivos con desarrollo de Cryptococcus sp., y a partir de éstos se obtuvo los registros de los pacientes, los que fueron analizados por dos revisores independientes. Resultados: Se recopiló la información de 13 pacientes, que presentaron 15 casos de infección por C. neoformans. De los 13 pacientes, 11 (84,6%) eran de sexo masculino, con una mediana de edad de 35 años. 11 pacientes (84,6%) padecían infección por VIH, uno (7,7%) tenía el antecedente de leucemia linfática crónica, y uno (7,7%) de etilismo crónico. De los 15 casos, nueve (60%) presentaron infección meníngea; cinco (33,3%) presentaron criptococcemia sin compromiso del LCR; y uno (6,6%) presentó infección pulmonar. De los 13 pacientes, ocho (53,3%) se encontraban fallecidos al año de seguimiento. Conclusiones: La infección por Cryptococcus sp. es una patología que debe ser sospechada en pacientes con inmunodeficiencia de predominio celular. La infección meníngea fue la forma más frecuente de presentación. Persiste presentando una elevada mortalidad.

Background: Cryptococcosis is a severe opportunistic mycotic infection, caused mainly by Cryptococcus neoformans. It can present as meningitis, pneumonia or cryptococcemia. Aim: To characterize patients with Cryptococcus infection between January 1°, 2013 and June 30, 2016, in Hospital Carlos van Buren, Valparaíso, Chile. Methods: We identified retrospectively those cultures with Cryptococcus sp. growth, and then obtained their clinical files which were analyzed by two independent reviewers. Results: We were able to obtain data from 13 of 15 patients who presented with Cryptococcus neoformans infection. Out of all, 11 (84.6%) were males, with a median age of 35 years old. 11 (84,6%) were HIV positive, 1 (7,7%) had chronic lymphocytic leukemia, and 1 (7,7%) refered alcohol abuse. Out of the 15 episodes, 9 (60%) had meningeal infection; 5 (33.3%) were cryptococcemia without meningeal involvement and 1 (6.6%) presented as a pulmonary infection. Eight patients were deceased at one year follow up. Conclusions: Cryptococcus sp. infection must be suspected in patients with cellular immunodeficiencies. Meningeal involvement is the most frequent form of clinical presentation. It still has a high mortality rate.

Caracterización molecular de los aislamientos de Cryptococcus neoformans de pacientes con HIV, Guayaquil, Ecuador / Molecular characterization of Cryptococcus neoformans isolates from HIV patients, Guayaquil, Ecuador

Resumen Introducción. La neurocriptococosis es una infección fúngica oportunista que representa un alto costo en vidas humanas y para la economía de los países. Sus agentes causales, las especies del complejo Cryptococcus neoformans/Cryptococcus gattii, tienen una fase sexuada y otra asexuada, cuatro serotipos principales y siete variedades moleculares con diferencias clínico-epidemiológicas, fenotípicas y de sensibilidad a los antifúngicos. Objetivo. Caracterizar molecularmente los aislamientos clínicos de C. neoformans de Guayaquil, Ecuador. Materiales y métodos. Se determinó el tipo de apareamiento, el serotipo y la variedad molecular mediante reacción en cadena de la polimerasa y análisis del polimorfismo de los fragmentos de restricción de 27 aislamientos levaduriformes previamente identificados como C. neoformans mediante métodos convencionales. Los aislamientos fueron recuperados del líquido cefalorraquídeo de pacientes con síndrome neurológico seropositivos para HIV, internados en el Hospital de Infectología "Dr. José Daniel Rodríguez Maridueña", entre diciembre de 2013 y enero de 2015. Resultados. Se demostró el amplio predominio de C. neoformans del serotipo A, MATα y el genotipo VNI entre los aislamientos estudiados. Conclusiones. Estos datos son similares a los obtenidos en otros países y son los primeros de su tipo en Guayaquil, Ecuador, por lo cual constituyen un aporte importante al conocimiento de la criptococosisen esta ciudad.

Abstract Introduction: Neurocryptococcosis is an opportunistic fungal infection that represents a high cost in human lives and for the economy of countries. Its causative agent, the Cryptococcus neoformans/Cryptococcus gattiispecies complex, has a sexual and an asexual phase, four major serotypes and seven molecular varieties with phenotypic, clinical-epidemiological and antifungal susceptibility differences. Objective: To characterize by molecular methods clinicalisolates of C. neoformans from Guayaquil, Ecuador. Materials and methods: We determined mating types, serotypes and molecular varieties by PCR and RFLP in 27 yeast isolates previously identified as C. neoformans by conventional methods. The isolates were recovered from cerebrospinal fluid of HIV seropositive patients with neurological syndrome admitted at "Dr. José Daniel Rodríguez Maridueña" Hospital from December, 2013, to January, 2015. Results: We established a wide prevalence of C. neoformans serotype A, MATαand genotype VNI among the studied isolates. Conclusions: These data are similar to those obtained in other countries and the first identified by molecular characterization in Guayaquil, Ecuador. Therefore, they constitute an important contribution to the knowledge on cryptococcosis in this country.

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

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

Disseminated cryptococcosis manifested as a single tumor in an immunocompetent patient, similar to the cutaneous primary forms

Abstract Cryptococcosis is a fungal infection caused by Cryptococcus neoformans that tends to affect immunocompromised individuals. The fungi are mostly acquired by inhalation, which leads to an initial pulmonary infection. Later, other organs - such as the central nervous system and the skin - can be affected by hematogenous spread. In addition, cutaneous contamination can occur by primary inoculation after injuries (primary cutaneous cryptococcosis), whose diagnosis is defined based on the absence of systemic involvement. The clinical presentation of cutaneous forms typically vary according to the infection mode. We report an unusual case of disseminated cryptococcosis in an immunocompetent patient with cutaneous lesions similar to those caused by primary inoculation. This clinical picture leads us to question the definition of primary cutaneous cryptococcosis established in the literature.

Distribution and association between environmental and clinical isolates of Cryptococcus neoformans in Bogotá-Colombia, 2012-2015

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.

First isolation of cryptococcus neoformans genotype VNI MAT-alpha from wood hollow trunks of hymenaea courbaril

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.

Aislamiento presuntivo y caracterización de cryptococcus neoformans y cryptococcus gattii desde árboles en la región de O’Higgins y Maule, Chile / Presumptive isolation and characterization of cryptococcus neoformans and cryptococcus gattii from trees in the region of O’Higgins and Maule, Chile

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 O’Higgins 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 O’Higgins 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.

Validation and clinical application of a molecular method for the identification of Cryptococcus neoformans/Cryptococcus gattii complex DNA in human clinical specimens

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.

Antifungal susceptibility of clinical and environmental Cryptococcus neoformans and Cryptococcus gattii isolates in Jabalpur, a city of Madhya Pradesh in Central India

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.

Cryptococcosis in Atlántico, Colombia: an approximation of the prevalence of this mycosis and the distribution of the etiological agent in the environment

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.

Isolation of cryptococcus neoformans from environmental samples collected in Southeastern Nigeria / Isolamento de Cryptococcus neoformans de amostras ambientais coletadas no sudeste da Nigéria

SUMMARYCryptococcosis caused by Cryptococcus neoformansis 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 neoformansor C. gattiiin 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 neoformanswas 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 neoformansfrom pigeon droppings in Nigeria. The ecological and epidemiological significance of these findings are discussed.

RESUMOA criptococose, causada por Cryptococcus neoformans, é o segundo patógeno fúngico oportunista mais comum em infecções com risco de vida e manifestações clínicas graves, especialmente em HIV/AIDS e outros pacientes imunocomprometidos. Na Nigéria, a infecção pelo HIV/AIDS atingiu um nível alarmante. Apesar disso, informações sobre a presença desses fungos em amostras clínicas e ambientais é muito escassa na Nigéria e em muitas outras partes da África. Propusemo-nos a avaliar a presença de Cryptococcus neoformansou C. gattiiem fezes de pombos obtidos do sudeste da Nigéria. Foram coletadas 177 amostras de fezes de pombos de seis localidades. A área coberta foi composta por dez cidades e outras localidades abrangendo cinco Estados na Nigéria. Usando técnicas estabelecidas, Cryptococcus neoformansfoi isolado de 39 do total de 177 (22,0%) amostras. Nenhuma amostra de C. gattiifoi isolada. A maioria dos isolados (32,4%) foi recuperada de pombais (11 de 34), seguido de perto por amostras colhidas em mercados (31,8%; 7 de 22) e por último na igreja (4,0%; 1 de 25). A maior taxa de isolamento (38,9%) foi encontrada em amostras de Enugu-Ezike (7 de 23), enquanto a menor taxa foi de Afikpo e os outros locais, cada um deles com taxa de isolamento de 9,1%. Esta é a primeira triagem em larga escala de Cryptococcus neoformansem fezes de pombos na Nigéria. A importância ecológica e epidemiológica destes achados é discutida.