Research progress of Cryptococcus gattii meningitis with different immune status / 中华神经科杂志

Cryptococcus gattii is a kind of Cryptococcus that infects the lungs and central nervous system through the inhalation of infectious particles such as spores or Cryptococcus yeast cells. The development of clinical disease of Cryptococcus gattii may be determined by the sex, immunity and genetics of the host factors, in which immune system factors play an important role in host injury. Their defects will have serious clinical consequences. Cryptococcus gattii mainly infects the population with normal immune, and the infection of immunosuppressed population is rare. The infection mechanism, molecular types, clinical characteristics, treatment and prognosis of Cryptococcus gattii meningitis were different between the two populations. This article reviews the main differences in different immune status with Cryptococcus gattii meningitis.

Clinical characteristics of cryptococcal meningitis patients with anti-granulocyte-macrophage colony-stimulating factor autoantibodies / 中华传染病杂志

Objective:To investigate the clinical characteristics and prognosis of cryptococcal meningitis patients with anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibodies.Methods:A total of 216 non-acquired immunodeficiency syndrome (AIDS) related cryptococcal meningitis cases with positive cultures of Cryptococcus, hospitalized at Huashan Hospital, Fudan University during January 2014 and December 2021, were retrospectively included. The serum anti-GM-CSF autoantibodies were detected by enzyme linked immunosorbent assay, and the clinical characteristics and prognosis were compared between patients with and without anti-GM-CSF autoantibodies. Statistical comparisons were mainly performed using the chi-square test or Fisher′s exact test. Cox proportional-hazards model was used to analyze the risk factors associated with prognosis. Results:Among 216 enrolled patients, 23 patients were positive of anti-GM-CSF autoantibodies, with a positive rate of 10.6%. Among 23 patients, seven cases were infected with Cryptococcus gattii, and 16 cases were infected with Cryptococcus neoformans. In the group with positive anti-GM-CSF autoantibodies, 30.4%(7/23) of the patients were infected with Cryptococcus gattii, which was higher than that of 1.6%(3/193) in the group with negative anti-GM-CSF autoantibodies, and the difference was statistically significant ( χ2=38.82, P<0.001). In the group with positive anti-GM-CSF autoantibodies, 30.0% (6/20) had mass lesions with a diameter greater than three centimeters in the lungs, and the one-year all-cause mortality rate was 50.0% (10/20), which were both higher than those of 3.4%(5/145) and 16.1% (29/180) in the negative group, respectively. The differences were both statistically significant (both Fisher′s exact test, P<0.01). Age≥60 years (hazard ratio ( HR)=4.146, P=0.002), predisposing factors ( HR=3.160, P=0.021), epilepsy ( HR=6.129, P=0.002), positive anti-GM-CSF autoantibodies ( HR=2.675, P=0.034), white blood cell count of cerebrospinal fluid (CSF)<100 ×10 6/L ( HR=2.736, P=0.039), the titers of cryptococcal capsular polysaccharide antigen of CSF≥1∶1 280 ( HR=4.361, P=0.009) were independent risk factors for one-year all-cause mortality in patients with cryptococcal meningitis. Conclusions:In non-AIDS related cryptococcal meningitis patients, the positive rate of serum anti-GM-CSF autoantibodies is as high as 10.6%. Patients with anti-GM-CSF autoantibodies could be infected with both Cryptococcus neoformans and Cryptococcus gattii, and they have higher proportion of lung mass lesions than patients with negative anti-GM-CSF autoantibodies. The one-year survival rate decreases significantly in patients with anti-GM-CSF autoantibodies, which is an independent risk factor for the prognosis of cryptococcal meningitis.

Criptococosis meníngea y pulmonar en paciente inmunocompetente

RESUMEN Presentamos el caso de un varón de 23 años, sin patologías conocidas, agricultor, procedente de la zona rural. Presenta un cuadro de 1 mes de evolución cefalea, náuseas y vómitos, además de alteración de la conciencia caracterizada por confusión. La tomografía de cráneo no muestra anomalías y la punción lumbar es compatible con meningitis, con tinta china positiva visualizándose Cryptococcus sp. Se inicia tratamiento con anfotericina B y fluconazol. Posteriormente retorna informe final de cultivo: Cryptococcus gatii. Se realizan estudios para descartar infecciones virósicas que retornan negativos. En su internación presenta neumonía nosocomial y por empeoramiento progresivo del estado de conciencia y la clínica respiratoria pasa a cuidados intensivos donde se mantuvo internado durante 21 días. En su estadía se confirmó además la presencia de C. gattii en lavado bronquioalveolar. Presentó falla multiorgánica y óbito. Es el primer caso de criptococosis en paciente inmunocompetente descrito en nuestro centro.

ABSTRACT We present the case of a 23-year-old male, with no known pathologies, a farmer, from a rural area. He presented a 1-month history of headache, nausea and vomiting, in addition to altered consciousness characterized by confusion. The skull tomography does not show abnormalities and the lumbar puncture is compatible with meningitis, with positive Chinese ink showing Cryptococcus sp. Treatment with amphotericin B and fluconazole is started. Later, the final culture report returns: Cryptococcus gatii. Studies are carried out to rule out viral infections that return negative. During his hospitalization, he presents nosocomial pneumonia and due to the progressive worsening of his state of consciousness and the respiratory symptoms goes to intensive care where he is hospitalized for 21 days. During his stay, the presence of C. gattii is also confirmed in bronchioalveolar lavage. He presents multiple organ failure and death. This is the first case of cryptococcosis in an immunocompetent patient described in our center.

Cryptococcosis by Cryptococcus neoformans/Cryptococcus gattii Species Complexes in non-HIV-Infected Patients in Southeastern Brazil

Abstract INTRODUCTION: The clinical manifestations of cryptococcosis are usually associated with the infecting agents Cryptococcus neoformans (CN) and C. gattii (CG) species complexes and the host. In this study, non-HIV-infected patients, at a university hospital in southeastern Brazil, had epidemiological and clinical data associated with cryptococcal disease and isolated Cryptococcus species: CN - 24 patients and CG - 12 patients. METHODS: The comparison was comprised of demographic data, predisposing factors, clinical and laboratory manifestations, and outcomes of cryptococcosis patients treated between 2000 and 2016. Immunocompetent and immunosuppressed patients were also compared, irrespective of the infecting species. Cryptococcus spp. were genotyped by PCR-RFLP analysis of the URA5 gene. RESULTS: Infections by the CN species complex (100% VNI genotype) were associated with drug immunosuppression and fungemia, and patients infected with the CG species complex (83% VG II and 17% VGI genotypes) had more evident environmental exposure and higher humoral response. CN and CG affected patients with or without comorbidities. CONCLUSIONS: Diabetes mellitus, other chronic non-infectious diseases, and alcoholism were likely predisposing factors for infection by both CN and CG species. Immunocompetent patients, independent of the infecting Cryptococcus species complexes, showed a higher occurrence of meningitis and a trend toward less fungal dissemination and longer survival than immunosuppressed hosts.

Frequency and geographical distribution of genotypes and mating types of Cryptococcus neoformans and Cryptococcus gattii species complexes in Argentina / Frecuencia y distribución geográfica de genotipos y tipos sexuales de los complejos de especies Cryptococcus neoformans y Cryptococcus gattii en Argentina

Abstract The aim of this work was to know the frequency and geographical distribution of genotypes and mating types of Cryptococcus neoformans and Cryptococcus gattii species complexes isolated from human infections in Argentina during the period from April 2009 to April 2011. A multicenter study was conducted, in which 372 isolates were obtained from 61 laboratories throughout the country. Of those, 98.8% of the isolates belonged to the C. neoformans species complex and 1.1% to the C. gattii species complex. Genotype VNI (MATa) was the most frequently isolated (n = 326, 87.6%), followed by VNII (MATa) (n = 22, 5.9%), the recently described VNII-VNIV (aADa) hybrid (n = 14, 3.8%), VNIV (MATa) (n=4, 1.1%), VNIII (aADa) hybrid (n = 1, 0.3%), and VNIII (aADa) hybrid (n = 1, 0.3%). The Argentine Central region showed the greatest number of cases and genotype diversity. Interestingly, a relative high frequency was observed in genotype VNII (MATa) in the Cuyo, Northeast and Northwest regions and, also in VNII-VNIV (aADa) hybrids in the Northwest region. C. gattii species complex was isolated at a low rate; 3 VGI (MATa) and 1 VGII (MATa) isolates were obtained from the Northwest and Central regions. In conclusion, this study shows that genotype frequencies seem to vary among regions in Argentina and reveals a relatively high frequency of rare hybrids in the Northwest region. Further regional clinical and environmental studies may help to elucidate if those varia-tions in frequencies are associated with the existence of regional ecological niches or any other regional factors.

Resumen El objetivo de! trabajo fue conocer la frecuencia y la distribución geográfica de genotipos y tipos sexuales de aislados pertenecientes a los complejos de especies Cryptococcus neoformansy Cryptococcus gattii obtenidos de infecciones humanas en Argentina. Entre abril de 2009 y abril de 2011 se realizó un estudio multicéntrico del que se obtuvieron 372 aislados de 61 laboratorios de diferentes zonas del país. El 98,8% de los aislados pertenecieron al complejo C. neoformansy el 1,1% al complejo C. gattii. El genotipo VNI (MATa) fue el más frecuente (n = 326; 87,6%), le siguieron VNII (MATa) (n = 22; 5,9%), el híbrido VNII-VNIV (aADa) (n = 14; 3,8%), VNIV (MATa) (n =4; 1,1%) y los híbridos VNIII (aADa) (n = 1; 0,3%) y VNIII (aADa) (n = 1; 0,3%). La región Centro mostró el mayor número de casos y la mayor diversidad de genotipos. Cabe destacar que el genotipo VNII (MATa) tuvo una frecuencia relativamente alta en las regiones de Cuyo, Noreste y Noroeste. En esta última región, también fue alta la frecuencia del híbrido VNII-VNIV (aADa). La frecuencia de aislamiento de miembros del complejo C. gattii fue baja: se obtuvieron 3 aislados VGI (MATa) y 1 VGII (MATa) de las regiones Centro y Noroeste. En conclusión, este estudio muestra que las frecuencias de genotipos varían entre las distintas regiones de Argentina y señala la presencia de híbridos poco comunes en una frecuencia relativamente alta dentro de la región Noroeste. Contar con mayor número de estudios clínicos y ambientales regionales podría ayudar a elucidar si tales variaciones están asociadas a la existencia de nichos ecológicos particulares o a algún otro factor regional.

Crecimiento del complejo Cryptococcus neoformans/ Cryptococcus gattiien extractos de excretas de palomas / Growth of Cryptococcus neoformans/Cryptococcus gattii species complex in pigeon excreta / Crescimento do complexo Cryptococcus neoformans/ Cryptococcus gattii em extratos de excrementos de pombos

RESUMEN Cryptococcus neoformans/Cryptococcus gattii son agentes etiológicos de la criptococosis, el primero tiene distribución mundial y su hábitat principal son las excretas de palomas (Columba livia), mientras que el último se ha aislado de árboles y en pocas ocasiones de heces de aves. En este estudio se comparó en condiciones de laboratorio, el crecimiento de C. neoformans y C. gattii en extractos de excretas de paloma. Se utilizaron dos extractos de heces de aves cautivas: uno de una muestra en la que se aisló C. neoformans (extracto positivo) y otro de una muestra negativa para este hongo (extracto negativo), los cuales fueron inoculados con C. neoformans y C. gattii. Los extractos permitieron el desarrollo de las dos especies y, mostraron un patrón de crecimiento uniforme en el extracto negativo, mientras que, en el positivo, el crecimiento de C. neoformans fue mayor comparado con el de C. gattii (P = 0.0001). El crecimiento de C. gattii fue menor a un pH superior a 7,5, mientras que el crecimiento de C. neoformans no mostró diferencias significativas en los dos extractos independiente de las variaciones del pH. Se necesitan más estudios para elucidar la interacción de estas dos especies en las excretas de palomas.

ABSTRACT Cryptococcus neoformans/Cryptococcus gattii are etiologic agents of cryptococcosis, the former has a worldwide distribution, and its habitat is associated with pigeon excreta (Columba livia), whereas the latter it has been mainly associated with trees and rarely from the feces of birds. In this study was to compare, under laboratory conditions, the growth of C. neoformans and C. gattii in extracts of pigeon excreta. Two extracts of captive bird feces were used: one prepared from a sample in which C. neoformans was isolated (positive extract) and one from which sample was negative for this fungus (negative extract). Under laboratory conditions, the extracts allowed the development of both species. Both showed a uniform pattern of growth in the negative extract, whereas in the positive, C. neoformans growth was significantly higher than that of C. gattii (P = 0.0001). The growth of the C. gattii was lower in positive extracts and it seems to decrease at pH greater than 7.5, whereas the growth of C. neoformans showed no significant differences regardless of pH variations. Further studies are need to elucidate the interaction between these two species in the excreta of pigeon.

RESUMO Cryptococcus neoformans / Cryptococcus gattii são agentes etiológicos da criptococose, o primeiro tem distribuição mundial e seu habitat principal é a excreção de pombos (Columba livia), enquanto o último foi isolado de árvores e raramente de fezes de aves. Neste estudo, o crescimento de C. neoformans e C. gattii nos extratos de excrementos de pombo foi comparado em condições de laboratório. Foram utilizados dois extratos de fezes de aves em cativeiro: um de uma amostra em que C. neoformans (extrato positivo) foi isolado e outro de uma amostra negativa para esse fungo (extrato negativo), que foi inoculado com C. neoformans e C gattii Os extratos permitiram o desenvolvimento das duas espécies e mostraram um padrão de crescimento uniforme no extrato negativo, enquanto no positivo o crescimento de C. neoformans foi maior em comparação ao de C. gattii (P = 0,0001). O crescimento de C. gattii foi menor a pH maior que 7,5, enquanto o crescimento de C. neoformans não mostrou diferenças significativas nos dois extratos, independentemente das variações de pH. Mais estudos são necessários para elucidar a interação dessas duas espécies nos excrementos de pombos.

Multilocus sequence typing (MLST) of clinical and environmental isolates of Cryptococcus neoformans and Cryptococcus gattii in six departments of Colombia reveals high genetic diversity

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.

Meningoencephalitis and optical neuritis caused by CRYPTOCOCCUS GATTII in an immunocompetent patient

The following case is of a 59-year-old man, undergoing no medication, with no pathological history or others risk factors, who presented dizziness, fever and asthenia twenty days before admission. The patient was admitted for investigation when the asthenia intensified, followed by seizures. On admission, blood count, biochemical tests and chest computed tomography were normal, a serological test for anti-HIV proved negative, while the magnetic resonance of the brain showed signs suggestive of meningoencephalitis. Cerebrospinal fluid (CSF) analysis suggested bacterial meningitis due to increased leukocytes with a predominance of polymorphonuclear cells, reduced glucose and increased proteins as well as positive Gram cocci in pairs by Gram and negative fungi by India ink test. Treatment with ceftriaxone was started. Since there was no significant improvement, CSF analysis was repeated on the seventh day of treatment. Intracranial pressure was measured by manometry (29 mmHg) and CSF analysis showed the presence of encapsulated yeasts similar to Cryptococcus neoformans by the India ink test. The treatment was modified to liposomal amphotericin B and flucytosine; the intracranial hypertension was controlled by repeated CSF punctures. After fourteen days of antifungal treatment, the patient presented visual turbidity and bilateral papillar edema, so corticosteroid therapy was prescribed. The evolution was favorable, with progressive resolution of symptoms, improvement of CSF parameters and visual acuity. The patient was discharged eight weeks after admission, with outpatient guidance. Corticosteroid therapy associated with antifungal therapy proved to be beneficial in this case, since following the introduction of corticosteroids there was progressive visual improvement.

Primer reporte de Cryptococcus gattii en Ecuador / First report of Cryptococcus gattii in Ecuador

El presente es el reporte de un caso de un paciente masculino de 37 años, con manifestaciones clínicas de aproximadamente 1 año de evolución caracterizadas por cefalea holocraneana de gran intensidad, fotofobia, diplopía, que se exacerban en los últimos tres meses con alucinaciones, pérdida de audición de oído izquierdo, refirió exposición de 4 años a heces de aves (palomas) por su trabajo en el puerto marítimo.Acudió por consulta privada a valoración y manejo, se le efectúa el diagnóstico de diabetes mellitus tipo 2, por lo cual se prescribió metformina oral, una vez iniciado el tratamiento presentó exacerbación de los síntomas neurológicos, por lo que se indicó internación hospitalaria y se sometió a investigación para diagnóstico de su enfermedad, con presunción de meningitis bacteriana, se le realizó punción lumbar para estudios microbiológicos y se inició tratamiento empírico con glucopeptido y carbapenemico. Con el esquema antimicrobiano presentó leve mejoría en la intensidad de sus síntomas, en el estudio microbiológico del líquido cefalorraquídeo se observaron estructuras micóticas compatibles con Cryptococcus, por lo cual se realizó una segunda punción lumbar para conseguir una muestra adecuada para cultivo y estudio con biología molecular, a fin de establecer el diagnóstico definitivo del microorganismo.

The present is a report of a 37-year-old male patient, who present clinical manifestations of approximately 1 year, by high-intensity holocranial headache, photophobia, diplopia, which were exacerbated in the last three months with hallucinations, loss of Hearing from the left ear, hereported exposure of 4 years to feces of birds (pigeons) for his work in the seaport.The patient was refer to a private consultation to assessment and management, he was diagnosed with type 2 diabetes mellitus, for which oral metformin was prescribed, once the treatment had begun, exacerbation of the neurological symptoms, so hospitalization was indicated, and He underwent research to diagnose his disease, with presumed bacterial meningitis, he underwent lumbar puncture for microbiological studies and began empirical treatment with glucopeptide and carbapenemic; With the antimicrobial scheme, there was a slight improvement in the intensity of its symptoms. In the microbiological study of the preliminary cerebrospinal fluid, fungal structures compatible with Cryptococcus were observe; a second lumbar puncture was performed to obtain an adequate sample for culture and study with molecular biology, in order to establish the definitive diagnosis of the microorganism.

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.

In vitro growth characteristics of Cryptococcus gattii VGⅠ/VGⅡ and their virulence diversity to Galleria mellonella / 中华微生物学和免疫学杂志

Objective@#To understand the in vitro growth characteristics of Cryptococcus gattii VGⅠ and VGⅡ isolated in China and the diversity in their virulence to Galleria mellonella.@*Methods@#Based on the results of multilocus sequence typing for eight strains of Cryptococcus gattii isolated in China, the strains were cultured in vitro to draw growth curves, observe the melanin production and measure the capsule thickness. The median lethal time (LT50) and median lethal dose (LC50) at 48 h of Cryptococcus gattii were calculated using Galleria mellonella infection test. Fourteen strains of Cryptococcus neoformans were studied for comparison.@*Results@#The eight Cryptococcus gattii strains were six VGⅠ and two VGⅡ. The growth curves of Cryptococcus gattii VGⅠ and VGⅡ were similar to that of Cryptococcus neoformans when culture at 30℃. The total number for each of them could reach 108 CFU/ml at 96 h under 30℃. However, the total number at any time point at 37℃ was less than that at 30℃. There was no significant difference in the amount of melanin produced by Cryptococcus neoformans under 30℃ and 37℃, but both VGⅠand VGⅡ types of Cryptococcus gattii could produce more amount of melanin under 37℃ than under 30℃. The ratio of capsule/cell wall diameter of Cryptococcus gattii VGⅠwas greater at 37℃ than that at 30℃ with statistical significance (P<0.001). Cryptococcus neoformans showed the longest LT50, followed by VGⅠand VGⅡ types of Cryptococcus gattii. The LT50 of Cryptococcus gattii VGⅡ at the concentration of 1×106 CFU/ml was 72 h, and its LC50 at 48 h was 1×108 CFU/ml.@*Conclusions@#Like Cryptococcus neoformans, Cryptococcus gattii VGⅠ and VGⅡ grew faster under 30℃ than under 37℃, but more melanin was produced and thicker capsule was formed under 37℃ than under 30℃. Among Cryptococcus neoformans and VGⅠ and VGⅡ types of Cryptococcus gattii, Cryptococcus gattii VGⅡ showed the shortest LT50 and the strongest virulence to Galleria mellonella.

In vitro growth characteristics of Cryptococcus gattii VGⅠ/ VGⅡ and their virulence diversity to Gal-leria mellonella / 中华微生物学和免疫学杂志

Objective To understand the in vitro growth characteristics of Cryptococcus gattii VGⅠ and VGⅡ isolated in China and the diversity in their virulence to Galleria mellonella. Methods Based on the results of multilocus sequence typing for eight strains of Cryptococcus gattii isolated in China, the strains were cultured in vitro to draw growth curves, observe the melanin production and measure the capsule thickness. The median lethal time (LT50 ) and median lethal dose (LC50 ) at 48 h of Cryptococcus gattii were calculated using Galleria mellonella infection test. Fourteen strains of Cryptococcus neoformans were studied for comparison. Results The eight Cryptococcus gattii strains were six VGⅠ and two VGⅡ. The growth curves of Cryptococcus gattii VGⅠ and VGⅡ were similar to that of Cryptococcus neoformans when culture at 30℃ . The total number for each of them could reach 108 CFU/ ml at 96 h under 30℃ . However, the total number at any time point at 37℃ was less than that at 30℃ . There was no significant difference in the amount of melanin produced by Cryptococcus neoformans under 30℃ and 37℃ , but both VGⅠand VGⅡtypes of Cryptococcus gattii could produce more amount of melanin under 37℃ than under 30℃ . The ratio of capsule/ cell wall diameter of Cryptococcus gattii VGⅠwas greater at 37℃ than that at 30℃ with statistical significance (P<0. 001). Cryptococcus neoformans showed the longest LT50 , followed by VGⅠand VGⅡtypes of Cryptococcus gattii. The LT50 of Cryptococcus gattii VGⅡ at the concentration of 1×106 CFU/ ml was 72 h, and its LC50 at 48 h was 1×108 CFU/ ml. Conclusions Like Cryptococcus neoformans, Cryptococcus gattii VGⅠ and VGⅡ grew faster under 30℃ than under 37℃ , but more melanin was produced and thicker capsule was formed under 37℃ than under 30℃ . Among Cryptococcus neoformans and VGⅠ and VGⅡtypes of Cryptococcus gattii, Cryptococcus gattii VGⅡ showed the shortest LT50 and the strongest virulence to Galleria mellonella.

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.

The status of cryptococcosis in Latin America

Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification of cryptococcal strains, in understanding their ecology, population genetics, host-pathogen interactions, and the clinical epidemiology of this important mycosis in Latin America.

Advances in Cryptococcus genomics: insights into the evolution of pathogenesis

Cryptococcus species are the causative agents of cryptococcal meningitis, a significant source of mortality in immunocompromised individuals. Initial work on the molecular epidemiology of this fungal pathogen utilized genotyping approaches to describe the genetic diversity and biogeography of two species, Cryptococcus neoformans and Cryptococcus gattii. Whole genome sequencing of representatives of both species resulted in reference assemblies enabling a wide array of downstream studies and genomic resources. With the increasing availability of whole genome sequencing, both species have now had hundreds of individual isolates sequenced, providing fine-scale insight into the evolution and diversification of Cryptococcus and allowing for the first genome-wide association studies to identify genetic variants associated with human virulence. Sequencing has also begun to examine the microevolution of isolates during prolonged infection and to identify variants specific to outbreak lineages, highlighting the potential role of hyper-mutation in evolving within short time scales. We can anticipate that further advances in sequencing technology and sequencing microbial genomes at scale, including metagenomics approaches, will continue to refine our view of how the evolution of Cryptococcus drives its success as a pathogen.

Fatal Cryptococcus gattii genotype VGI infection in an HIV-positive patient in Barranquilla, Colombia

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.

Equine nasopharyngeal cryptococcoma due to Cryptococcus gattii / Criptocoma nasofaríngeo equino devido a Cryptococcus gattii

ABSTRACT: Cryptococcus gattii is often associated with pulmonary and systemic infections in humans and animals. In this research we report a case of nasopharyngeal cryptococoma caused by C. gatti in an equine. A 10-year-old mare presented a mass obstructing the oropharynx. Macroscopically the mass was asymmetric, and was attached to the ethmoidal sinuses and obstructed the oropharynx. Histopathological examination of the mass revealed multiple yeast cells ranging from spherical to oval, 4-8μm in diameter, with some of them showing narrow base polar budding. Cryptococcus gattii growth in mycological culture (Sabouraud Dextrose Agar) and was L-canavanine-glycine-bromothymol blue Agar positive. The molecular identification confirmed the isolate as C. gattii by means of the amplification of universal primers. C. gattii is considered an emerging fungal agent, as it affects human and animals and does not respond efficiently to commonly established treatments.

RESUMO: Cryptococcus gattii é frequentemente associada a infecções pulmonares e sistêmicas em humanos e animais. Neste relato descreve-se um caso de criptococoma nasofaríngeo por C. gatti em um equino. Uma égua de 10 anos, apresentou uma massa obstruindo a orofaringe. Macroscopicamente a massa era assimétrica, e estava aderida aos seios etmoidais obstruindo a orofaringe. O exame histopatológico da massa revelou múltiplas células leveduriformes variando de esféricas a ovais, de 4-8μm de diâmetro, com algumas delas apresentando brotação polar de base estreita. Cryptococcus gattii foi isolado na cultura micológica (Ágar sabouraud dextrose) e foi positivo no Ágar L-canavanina-glicina-azul de bromotimol. A identificação molecular confirmou o isolado como C. gattii por meio da amplificação de primers universais. O diagnóstico de criptococoma por C. gattii nasofaríngeo foi baseado nos sinais clínicos, achados macroscópicos, lesões histológicas, cultura micológica e Reação em Cadeia da Polimerase. O C. gattii é considerado um agente fúngico emergente, pois acometendo humanos e animais, não respondendo com eficiência aos tratamentos comumente estabelecidos.

Mechanistic study on the role of microRNA-146a in THP-1 cells-associated inflammatory response induced by Cryptococcus neoformans and Cryptococcus gattii / 中国感染与化疗杂志

Objective To analyze the differential expression of microRNA-146a (miR-146a) in monocyte-macrophage cell line (THP-1 cells) after induction by Cryptococcus neoformans (C.neoformans,reference strain WM148) or Cryptococcus gattii (C.gattii,reference strain R265),and investigate the mechanism of miR-146a in regulating the inflammatory response of cryptococcal meningitis.Methods The cultured THP-1 cells were divided into two groups to be induced by C.neoformans or C.gattii,respectively.THP-1 cells were induced with inactivated WN148 (or R265) strains at multiplicity of infection (MOI) of 5 in all experiments.The supematant and the cell pellet were collected separately after incubation.The expression of miR-146a was measured by real-time quantitative PCR (qRT-PCR) technique.The levels of TNF-u and IL-6 release were assayed by ELISA.Results The expression of miR-146a increased significantly in the C.neoformans induction group compared to 0 h.It reached peak at 3 h (P＜0.01),and then declined gradually.The level of TNF-α increased in supematant and reached peak at 12 h.The expression of IL-6 did not change significantly at each time point.The expression of miR-146a and TNF-α increased gradually and reached peak at 12 h in the C.gattii induction group (P ＜0.01),but the change did not reach statistical significance at 3 h,6 h time points.The expression of IL-6 gradually increased,and reached peak at 12 h time point.Conclusions Following stimulation with C.neoformans or C.gattii,the expression ofmiR-146a in THP-1 cells showed different patterns over time.The expression levels of TNF-α and IL-6 showed different patterns.These findings suggest that there may be different regulatory mechanisms in the THP-1 cells-associated inflammatory response after stimulation by inactivated C.neoformans and C.gattii strains.

Disseminated cryptococcosis with cutaneous involvement in an immunocompetent patient

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.

Distribuição de tipos moleculares de Cryptococcus gattii no Brasil: uma revisão bibliográfica / Distribution of Cryptococcus gattii molecular types in Brazil: a bibliographic review

Objetivo: descrever a distribuição molecular do Cryptococcus gattii no Brasil por meio de revisão de estudos publicados até 2016. Fonte de dados: Revisão bibliográfica de estudos publicados até 2016 no sítio DeCS BVS com os descritores nos idiomas português, Inglês e espanhol. O critério de inclusão foi: abordar resultados de caracterização molecular de C. gattii no Brasil. Critérios de exclusão: artigos sem texto completo e artigos sem informações moleculares de C. gattii. Síntese de dados: Inclui se dez publicações entre 2008 e 2016 que contêm informações sobre a caracterização molecular de C. gattii de isolados encontrados nas quatro regiões do Brasil. Conclusões: C. gattii é endêmico nas regiões Norte e Nordeste do Brasil, mas pode ser encontrado em fontes primárias nas demais regiões. O tipo molecular mais predominante foi o VGII, responsável por cerca de 80% dos casos. VGII apresentou uma alta variabilidade genética. O VGII encontrado na região Nordeste é diferente do encontrado na região Norte.

Objective: describe the distribution of Cryptococcus gattii molecular types in Brazil by reviewing studies published until 2016. Source of data: Review of studies published until 2016 in the DeCS BVS site in Portuguese, English or Spanish. The inclusion criterion was description of molecular typing of C. gattii isolates obtained in Brazil. Manuscripts without full text and without molecular information on C. gattii were excluded. Synthesis of the collected data: We have included ten publications ranging from 2008 to 2016 containing information about the molecular characterization of C. gattii isolates from the four geographic regions of Brazil. Conclusions: C. gattii is endemic in the North and Northeast regions of Brazil, but can also be found in the other regions. The predominant molecular type was VGII, accounting for about 80% of the cases. VGII has very high genetic variability. VGII isolates from the Northeast region are distinct from those found in the North.