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.

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.

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.

First case report of cryptococcosis due to Cryptococcus decagattii in a pediatric patient in Argentina

Abstract We report the first case of cryptococcosis due to Cryptococcus decagattii in an immunocompetent pediatric patient from an indigenous community in Argentina with a successful outcome. Two isolates (blood, cerebrospinal fluid) were genotyped by restriction fragment length polymorphism of the orotidine monophosphate pyrophosphorylase (URA5) gene as VGIV and identified by multi-locus sequence typing as C. decagattii. Matrix-assisted laser desorption/ionization time of flight mass spectrometry identification indicated genotype VGIII. The minimum inhibitory concentration of amphotericin B, fluconazole, itraconazole, and voriconazole was determined (cerebrospinal fluid: 0.25, 16, 0.12, and 0.12, blood: 0.25, 4, 0.12, and 0.06, respectively, all in mg/L).

Diagnóstico de Cryptococcus neoformans mediante la PCR- RFLP en el líquido cefalorraquídeo (LCR) en pacientes hospitalizados en el sur de Brasil / Cryptococcus neoformans diagnosis using PCR-RFLP on cerebrospinal fluid (CSF) of hospitalized patients in southern Brazil

Introducción: Cryptococcus neoformans es un hongo levaduriforme encapsulado, de distribución mundial, principalmente en regiones tropicales, causando infecciones en individuos inmunocomprometidos, sobre todo en los infectados con el virus de la inmunodeficiencia humana (HIV). La capacidad de infección de este hongo es variable, pudiendo citar la facultativa patogenicidad, cápsula con actividad fagocitaria y producción de melanina como antioxidante. El objetivo de este trabajo fue evaluar el uso de la PCR/ RFLP para la detección e identificación de C. neoformans directamente del líquido cefalorraquídeo (LCR) de pacientes ingresados en un hospital público de la ciudad de Lages, Estado de Santa Catarina, sur de Brasil, comparando el resultado con la tinción específica para el hongo y el crecimiento en medio de cultivo. Métodos: Las muestras fueran directamente teñidas con tinta china para observar la cápsula, bien como después sembladas en medio de cultivo (agar dextrosa Sabouraud y agar de Níger) para crecimiento fúngico; también se hizo la extracción del ADN con fenol-cloroformo. La técnica fue utilizada para amplificación del gen URA5 mediante reacción en cadena de la polimerasa (PCR) y después con las enzimas de restricción HhaI y Sau96I para genotipaje mediante la PCR-RFLP. Resultados: En dos muestras fueran aislados C. neoformans con la tinción china y amplificados en la PCR, en las cuales fueran identificados como var. grubii. Conclusión: El serotipo A var. grubii es lo más aislado en la criptococosis humana, principalmente en pacientes HIV, pero se desconoce la preferencia de este serotipo por este grupo de enfermos. (AU)

Introduction: Cryptococcus neoformans is an encapsulated yeast with worldwide circulation, predominantly in tropical regions, causing infections in immunocompromised individuals, particularly those infected with human immunodeficiency virus (HIV). The virulence of this fungus is variable, and it should be mentioned the facultative pathogenicity, capsule with anti-phagocytic activity, and antioxidant melanin production. The aim of this study was to evaluate the use of polymerase chain reaction (PCR)/ restriction fragment length polymorphism (RFLP) for detection and identification of C. neoformans directly from the cerebrospinal fluid (CSF) of patients admitted to a public hospital with suspected meningitis and/or meningoencephalitis in the city of Lages, Santa Catarina, southern Brazil. Results were compared using Chinese ink and growth media. Methods: The samples were submitted to direct examination with Chinese ink for capsule observation, then to growth on culture media (Sabouraud Agar and Niger), with subsequent DNA extraction with phenol-chloroform. PCR was the technique used for amplification of URA5 gene, and then restriction enzymes HhaI and Sau96I were used for genotyping by PCR-RFLP. Results: In two samples, C. neoformans were isolated by Chinese ink and amplified by PCR. They were identified as serotype var. grubii. Conclusion: C. neoformans var. grubii is the most commonly isolated in human cryptococcosis, mainly in HIV patients. However, the preference of this serotype for this group of patients is unknown. (AU)

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.

Essential oils and major compounds of Hedychium coronarium Koenig (Zingiberaceae) against pathogenic yeast of Candida and Cryptococcus genus / Óleos essenciais e compostos majoritários de Hedychium coronarium Koenig (Zingiberaceae) contra leveduras patogênicas dos gêneros Candida e Cryptococcus

Among the major causative agents of invasive fungal infections stands out the opportunistic yeasts of Candida and Cryptococcus. Regarding the problem of the high incidence of infections by these agents and the difculty of treating the low stockpile of antifungal drugs and the high toxicity of most therapies, the search for new antifungal compounds has been highlighted in recent decades. Hedychium coronarium, popularly known as "lírio-do-brejo" or "gengibre-branco" features several previously reported biological activities, including antimicrobial activity. Compound 1.8-cineole is the major compound in essential oil extracted from roots of H. coronarium, while caryophyllene oxide presents itself as the major in essential oil extracted from leaves of this plant. Our data show strong antifungal activity of compounds, against species of Candida albicans, Candida parapsilosis, Candida krusei, Cryptococcus neoformans and Cryptococcus gattii, with minimal inhibitory concentration and minimal fungicidal concentration equal to 0.2 % (v/v) for essential oil extracted from roots, while the essential oil extracted from leaves showed no activity against yeasts. The caryophyllene oxide showed higher antifungal activity for Cryptococcus spp. Thus, our results showed that the essential oil of rhizome is a promising antifungal agent against pathogenic yeasts.(AU)

Candida spp e Cryptococcus spp estão classifcadas entre os maiores causadores de infecções fúngicas invasivas em pacientes imunocomprometidos. Diante a alta incidência destas infecções por estes agentes e a difculdade do sucesso no tratamento, decorrente do baixo arsenal de fármacos antifúngicos e da alta toxicidade presente na maioria dos esquemas terapêuticos, a busca por novos compostos antifúngicos tem sido alvo de diversos estudos nas últimas décadas. Hedychium coronarium, popularmente conhecido como "lírio-do-brejo" ou "gengibre-branco", apresenta diversas atividades biológicas já descritas, entre elas a atividade antimicrobiana. O composto 1.8-Cineol é o composto majoritário presente no óleo essencial extraído de raízes de H. coronarium e o composto óxido de cariofleno é o composto majoritário extraído das folhas desta planta. Nossos resultados mostram que os compostos extraídos de H. coronarium apresentam forte atividade contra Candida albicans, Candida parapsilosis, Candida krusei, Cryptococcus neoformans e Cryptococcus gattii, com valores de concentração inibitória minima e concentração fungicida minima igual a 0,2 % (v/v) para o óleo essencial extraído das raízes, enquanto que o óleo essencial extraído das folhas, não mostrou atividade contras as leveduras. O composto óxido de cariofleno mostrou maior atividade antifúngica para Crytopcoccus spp. Assim, nossos dados mostraram que o óleo essencial extraído das raízes de H. coronarium, é um agente antifúngico promissor contra leveduras patogênicas.(AU)

Cryptococcus laurentii fungaemia in a cervical cancer patient

ABSTRACT Infections caused by emerging Cryptococcus non-neoformans species are being reported with increasingly frequency. Here, we present a case of fungaemia byCryptococcus laurentii in a woman receiving aggressive immunosuppressive therapy for cervical neoplasia. Three venous blood samples were aseptically collected on consecutive days and C. laurentiiwas isolated and identified through phenotypic and molecular methods. After central venous catheter removal and appropriate antifungal therapy, the patient showed significant improvement and blood culture became negative. Thus, patients following immunosuppressive therapies and using invasive medical devices are at risk of C. laurentii blood infections.

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 Colombia: Results of the national surveillance program for the years 2006-2010 / Criptococosis en Colombia: resultados de la encuesta nacional, 2006-2010

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.

Posterior fossa midline cryptococcoma in a patient with idiopathic CD4 lymphocytopenia.

Idiopathic CD4 lymphocytopenia (ICL) is a rare disorder which is often diagnosed as HIV-negative AIDS in the light of poor immunity and AIDS-defining illnesses. We present a case of a 50-year-old male who presented with a midline posterior fossa tumour with ICL diagnosed as cerebellar cryptococcoma.

Vertebro-cerebral cryptococcosis mimicking tuberculosis: A diagnostic dilemma in countries with high burden of tuberculosis.

We report a case of a 30-year-old immunocompetent man with disseminated cryptococcosis who was initially treated with antitubercular therapy due to clinical and radiological diagnosis of vertebro-cerebral tuberculosis. The diagnosis of Cryptococcus infection was made due to incidental isolation of this fungus from blood culture with negative cerebrospinal fluid culture results. Though disseminated cryptococcosis with central nervous system, skeletal, and skin involvement is an uncommon manifestation of Cryptococcus neoformans infection, a high clinical suspicion and early initiation of therapy is needed to recognise and treat such patients efficiently.

Antifungal susceptibilities of Cryptococcus species complex isolates from AIDS and non-AIDS patients in Southeast China

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.

Isolation of cryptococcus spp. in excrements of pigeons (columba sp.) in the Maringá city, Pr, Brazil / Isolamento de cryptococcus spp. em excretas de pombos (columba sp.) na cidade de Maringá, Pr, Brasil

The Cryptococcus genus is composed by 37 different species, however, the C. gattii and C. neoformans are the two most concerning species once they cause deep mycosis in humans. In this way, the present study had as aim to perform an analysis in droppings of pigeon (Columba sp.) from the city of Maringá, PR, in order to verify the presence of suggestive yeast forms of Cryptococcus spp. For this study 70 samples of pigeon excrements were evaluated which were collected in 7 different sites. These samples were submitted to the tests of thermotolerance at 37°C, urease, phenoloxidase activity and direct exam with China ink preparation. The results show that the pigeon droppings of 5,7% of the studied samples presented suggestive yeast forms of Cryptococcus spp. These results are relevant due to the positive samples are from the sites with a higher possibility of immunocompromised individuals flow, who are more up to have the most severe kinds of the disease caused by this fungi.

O gênero Cryptococcus é composto de 37 espécies diferentes, contudo o C. gattii e C. neoformans são as mais preocupantes por causarem micose profunda em humanos. Desta forma, o presente trabalho teve como objetivo realizar uma análise em excretas de pombos (Columba sp.) procedentes da cidade de Maringá, PR, para verificar a presença de leveduras sugestivas de Cryptococcus spp. Para este estudo foram analisadas 70 amostras de excretas de pombos, coletadas em sete localidades diferentes. Essas amostras foram submetidas ao teste de termotolerância à 37°C, urease, atividade fenoloxidase e exame direto com tinta da china. Os resultados demonstraram que nos excrementos de pombos de 5,7% das localidades estudadas apresentaram leveduras sugestivas de Cryptococcus spp. Estes resultados mostraram-se relevantes, devido ao fato de as amostras positivas pertencerem aos locais de maior possibilidade de circulação de indivíduos imunocomprometidos, os quais estão mais propícios a contrair as formas mais graves da doença causada por este fungo.

Unusual morphologies of Cryptococcus spp. in tissue specimens: report of 10 cases / Histopatologia, sorologia e cultivo no diagnóstico da criptococose

Ten cases of cryptococcosis due to unusual microscopic forms of Cryptococcus sp. observed over a twenty-eight year period (1981-2009) are presented. The most important clinicopathological and laboratory data are tabulated. The uncommon forms of cryptococcal cells given are: structures resembling germ tube (one case), chains of budding yeasts (one case), pseudohyphae (two cases) and nonencapsulated yeast-like organisms (eight cases). The diagnosis was based on the histopathological findings. The causative organism was isolated and identified in seven cases; five were due to C. neoformans, and two to C. gattii. In addition, the importance of using staining histochemical techniques - Grocott's silver stain (GMS), Mayer's mucicarmine stain (MM) and Fontana-Masson stain (FM) - in the diagnosis of cryptococcosis is argued.

A criptococose é a mais comum infecção fúngica oportunística observada em pacientes com síndrome da imunodeficiência adquirida (AIDS). Relatamos 13 casos da infecção baseados no diagnóstico histopatológico, sorológico e cultivo. Foram analisadas: a epidemiologia, as técnicas histoquímicas básicas de hematoxilina-eosina (HE) e coloração pela prata (GMS), bem como as técnicas histoquímicas especiais de mucicarmim de Mayer (MM) e Fontana-Masson (FM), o teste do antígeno criptocóccico (CrAg) e o isolamento em cultivos em ágar-Sabouraud (SAB), ágar infusão de cérebro-coração (BHI) e meio com canavanina azul de bromotimol (CGB). Em quatro casos, resultados tintoriais insatisfatórios pela coloração de MM associados a títulos negativos pelo teste do CrAg, a coloração de FM confirmou a infecção pelo Cryptococcus deficiente de cápsula. Oito isolados foram identificados: seis casos apresentaram a infecção por Cryptococcus neoformans e dois casos apresentaram a infecção por Cryptococcus gattii.