Study of Cryptococcus neoformans Varieties

Summary: We recommend L-canavanine-glycine bromothymol blue medium to differentiate C. neoformans var. neoformans from C. neoformans var. gattii. The former was isolated both from saprophytic and parasitic sources, the latter variety (7 isolates) being only isolated from clinical cases. One strain came from China, and six from Zaïre where they were isolated before 1969.
Zusammenfassung: Zur Unterscheidung zwischen C. neoformans var. neoformans und C. neoformans var. gattii wird das L-Canavanine-Glycin-Bromothymolblau-Medium empfohlen. C. neoformans var. gattii wurde nur von klinischen Fällen isoliert: ein Stamm aus China und sechs Stämme aus Zaïre. Alle Stämme aus Zaïre wurden vor 1969 isoliert.     

Osteomyelitis durch Cryptococcus neoformans

Zusammenfassung. Ein 43jähriger Patient kam zur stationären Aufnahme mit der Verdachtsdiagnose Knochentumor im linken Oberschenkel, stattdessen wurde Cryptococcus neoformans als Erreger einer Osteomyelitis kulturell nachgewiesen. Anamnestisch war als prädisponierender Faktor eine Corticosteroiddauertherapie aufgrund einer Sarkoidose bekannt. Die Therapie der Osteomyelitis erfolgte mit Fluconazol (200 mg/d p.o.) über 3 Monate. Darunter wurde der Patient beschwerdefrei.
Summary. A fortythree-year old patient suspected of having a bone tumor in his left femur, was submitted to the hospital. However, osteomyelitis caused by Cryptococcus neoformans has been demonstrated by culture. The history of the patient revealed a long-term therapy with corticosteroids due to sarcoidosis. The osteomyelitis was treated with fluconazole (200 mg daily p.o) for three months. Under this therapy the infection resolved.     

Infection due to Cryptococcus neoformans of unusual morphology in a patient with AIDS

Cryptococcus neoformans with a rare morphology of hand-mirror appearance was demonstrated by direct microscopic preparation of both cerebrospinal fluid (CSF) and sputum of a patient with AIDS. In addition, one to six blastoconidia were seen at the tip of a germ-tube like structure. Cr. neoformans was isolated in pure culture and the identification was confirmed by biochemical and serological tests as well as by animal pathogenicity.     

Isolation of Cryptococcus species including C. neoformans from cloaca of pigeons

Upper digestive tract of the pigeon (Columba livia) is well known as a reservoir for different species of Cryptococcus, but lower portions are not so frequently studied. In the present study, we tested on selective media a total of 331 pigeon cloacal swabs; Cryptococcus spp. were recovered from 26 (7.85%). Cryptococcus uniguttulatus was isolated from 11 samples (3.32%), C. laurentii from six (1.81%), C. neoformans var. neoformans from six (1.81%) and C. albidus from three of them (0.91%). The results show the importance of pigeon in the cryptococcosis epidemiology as reservoir and carrier for C. neoformans var. neoformans, but also for other Cryptococcus species of increasing clinical interest.     

First isolation of Cryptococcus neoformans var. gattii from a native jungle tree in the Brazilian Amazon rainforest

Cryptococcus neoformans var. gattii was isolated for the first time from decaying wood in a hollow of a native jungle tree Guettarda acreana, in a wild area of an Amazon rainforest island, in Brazil. The presence of this variety in a virgin environment without either anthropic action or introduced vegetation is discussed with regard to the common knowledge of Cr. neoformans ecology.     

Detection of Cryptococcus neoformans in Biopsy Specimens from the Spleen and the Liver of AIDS Patients: Critical Comments: Nachweis von Cryptococcus neoformans in Biopsiematerial von Milz und Leber AIDS-Kranker: Eine kritische Stellungnahme

Summary: This article was prompted by a publication describing the use of fluorochrome for rapid detection of Cryptococcus neoformans in the spleen of an AIDS patient In two AIDS patients including the above-mentioned one, cryptococcosis was diagnosed by the author on the basis of biopsy specimens from spleen and liver. The rapid microscopic diagnosis of the pathogen in histological sections stained with HE and PAS as well as in the unstained native preparations was without problems; for this reason, additional methods were found to be unnecessary. Subsequently, in both cases a disseminated cryptococcosis was confirmed miroscopically, culturally and serologically. It is pointed out that for the diagnosis of cryptococcosis in AIDS patients, invasive methods such as spleen and liver biopsy should not be used but recognized cultural and serological methods at an early stage of the fungal infection. Nevertheless, for such mycological diagnosis, special knowledge of differentiating methods is required. Attempts of performing a rapid diagnosis using fluorescence microscopy are considered as dispensable. Zusammenfassung: Anlaß zu vorliegendem Beitrag war eine Veröffentlichung in dieser Zeitschrift, in der die Verwendung von Fluorochrom zur Schnelldiagnose einer Cryptococcus-neoformans-Infektion in der Milz eines AIDS-Patienten, die bereits vom Autor gestellt worden war, beschrieben wurde. Der Autor konnte in Milz- und Leberstanzzylindern von diesem und beim Leberstanzzylinder eines weiteren AIDS-Patienten aus derselben Klinik eine Cryptococcusneoformans-Infektion nachweisen. Die mikroskopische Schnelldiagnose des Erregers in mit HE und PAS angefárbten histologischen Schnitten wie auch in ungefärbten Nativpräparaten bot keine Schwierigkeiten; somit erübrigte sich die Anwendung weiterer mikroskopisch-diagnostischer Verfahren. In beiden Fällen wurde mikroskopisch, kulturell und serologisch eine disseminierte Kryptokokkose festgestellt Invasive Verfahren wie Milz-und Leberpunktion sind somit entbehrlich, denn bewährte kulturelle und serologische Verfahren ermöglichen eine Schnelldiagnose bereits im Frühstadium der Infektion.     

In vitro susceptibilities of Malaysian clinical isolates of Cryptococcus neoformans var. grubii and Cryptococcus gattii to five antifungal drugs

The in vitro susceptibilities of Malaysian clinical isolates of Cryptococcus neoformans var. grubii and C . gattii to five antifungal drugs (amphotericin B, flucytosine, fluconazole, itraconazole and ketoconazole) were determined using the Etest method. None of the Malaysian isolates was resistant to amphotericin B and ketoconazole. Isolates resistant to flucytosine, fluconazole and itraconazole were observed in this study. Minimum inhibition concentrations (MICs) of > or = 32 microg ml(-1) against flucytosine, > or = 64 microg ml(-1) against fluconazole and > or = 1 microg ml(-1) against itraconazole were noted in four (8.3%), two (4.2%) and one (2.1%) isolates respectively. There was no significant difference in the MICs for both Cryptococcus species (P > 0.05), indicating that C. gattii was as susceptible as var. grubii to all the antifungal drugs tested. No significant difference in the MICs for both Cryptococcus species collected from 1980 to 1990 and 2002 to 2004 were observed (P > 0.05).     

Tödliche Meningoenzephalitis durch Cryptococcus neoformans var. neoformans bei einem Mädchen ohne schwerwiegende Immundefekte

Zusammenfassung. Es wird der Verlauf einer Meningoenzephalitis durch Cryptococcus neoformans bei einer 14jährigen Patientin dargelegt. Die mykologischen Untersuchungsbefunde wiesen anfangs einen massiven Befall des Liquors mit Cryptococcus neoformans (ca. 10000 Hefezellen/ml) und wochenlang sehr hohe Cryptococcus -Antigentiter (maximal 1:2048) aus. Eine HIV-Infektion oder andere schwerwiegende Immundefekte konnten ausgeschlossen werden. Die Behandlung der Cryptococcose erfolgte mit der Dreifachkombination Amphotericin B, Flucytosin und Fluconazol. Der Liquor konnte saniert werden. Trotzdem kam es zu erheblichen Läsionen am Gehirn. Die Patientin ist an den Folgen der Pilzinfektion am 74. Behandlungstag verstorben. Der geschilderte Krankheitsverlauf ist Anlaß, auf die Differentialdiagnose, Cryptococcus neoformans -Infektion"bei der chronischen lymphozytären Liquorpleozytose, verbunden mit Hirndruck- und Augensymptomen, hinzuweisen.
Summary. Case report on a lethal meningo-encephalitis due to Cryptococcus neoformans in a 14-year-old girl without serious immunodeficiency inclusive HIV-infection. The detection of high quantities of cells of Cryptococcus neoformans (about 10000/ml) and high levels of Cryptococcus antigen (up to 1:2048) in the cerebrospinal fluid are remarkable. The patient was treated with a triple combination of amphotericin B, flucytosine and fluconazole. After 18 days the cerebrospinal fluid was sterile. Nevertheless considerable lesions of the brain arised. The patient died from the Cryptococcus infection on day 74 of the antimycotic therapy. Cryptococcosis should be included into the differential diagnosis of the chronic lymphocytic pleocytosis of the cerebrospinal fluid connected with symptoms of intracranial pressure and ocular symptoms.     

Detection of Cryptococcus neoformans var. grubii in honeybee (Apis mellifera) colonies

The plant flora has an important role in the ecology of Cryptococcus neoformans. It is estimated that the environmental spreading and contamination of human beings with this yeast occurs via contaminated particles of plants. Cultivation of canopy parts of plants in selective media is the most widely used isolation method of this yeast. Cryptococcus neoformans var. grubii was isolated from honeybee colonies in Eucalyptus forests but was not isolated from the places where this flora did not exist. Our results indicate that the occurrence of C. neoformans in honeybee colonies during the flowering season of Eucalyptus spp. trees can be an important bioindicator for environmental yeast presence. The screening of honeybee colonies is a practical and a rapid method for the monitoring of the C. neoformans presence in flowering plants.     

Proven pulmonary cryptococcosis due to capsule-deficient Cryptococcus neoformans does not differ clinically from proven pulmonary cryptococcosis due to capsule-intact Cr. neoformans

We compared the clinical manifestations, laboratory and radiologic findings, and outcomes of eight patients with proven pulmonary cryptococcosis (PC) caused by capsule-deficient Cryptococcus neoformans with those of six patients with PC caused by capsule-intact Cr. neoformans. The presentations and outcomes did not differ significantly between the groups.     

Immune reconstitution inflammatory syndrome (IRIS) associated with Cryptococcus neoformans infection in AIDS patients

Cryptococcosis is frequently associated to the immune reconstitution inflammatory syndrome (IRIS) in AIDS patients on highly active antiretroviral therapy (HAART). This study aimed to evaluate clinical and evolutive features of IRIS associated cryptococcosis patients in Uberaba, Brazil. Patients: Eighty‐one AIDS individuals admitted at the teaching hospital with cryptococcal meningitis were evaluated and from these, 40 were prospectively followed. Of 40 patients with cryptococcosis, nine (22.5%) presented clinical and laboratory features of IRIS. Six (66.6%) were male, with a mean age of 37.2. Five (55.5%) presented cryptococcosis as first AIDS defining condition. In seven (77.9%) IRIS was characterised as a relapse of meningeal symptoms after 10 weeks, mean time of 72 days, of starting HAART whereas, two asymptomatic patients developed the syndrome as an unmasked cryptococcosis after 10 and 12 weeks on HAART. Lymphadenitis as isolated finding associated with IRIS was evidenced in three cases. All patients presented low CD4⁺ and high RNA viral load baseline values. Cultures of cerebrospinal fluid and lymph‐node fragments tissues of these cases were negative. Six of nine individuals developed high intracranial pressure requiring a daily relief lumbar puncture. No deaths occurred during the evolution of these patients. The incidence and clinical evolutive profile observed in this case series are in accordance with other reports elsewhere.     

Schleimkapselbildung in vitro von Cryptococcus neoformans: Mucoid Encapsulation in vitro of Cryptococcus neoformans

Zusammenfassung:  Die Schleimkapselbildung in vivo von Cryptococcus neoformans ermöglicht häufig das Erkennen einer Cryptococcosis bereits bei der Direktuntersuchung (Liquor cerebrospinalis, histologische Präparate). Auf herkömmlichen festen oder flüssigen Medien bleibt die Kapselbildung zumeist aus. Deshalb wurde ein neues Medium formuliert, das die Kapselbildung besonders begünstigt. Hierdurch wird die Bestimmung von Cryptococcus neoformans erleichtert.
Summary:  The formation of a mucoid capsule of Cryptococcus neoformans cells in vivo often is helpful in the histological diagnosis of a cryptococcosis or when samples of cerebrospinal fluid are studied by direct microscopy. In cultures on conventional solid or liquid media encapsulation frequently does not occur. For this reason, a new medium has been developed which stimulates capsule formation and thus facilitates the identification of Cryptococcus neoformans.     

Isolation of Cryptococcus neoformans van neoformans in the Aviaries of the Antwerp Zoological Gardens/Isolation von Cryptococcus neoformans van neoformans im Vogelhaus des Antwerpener Zoos

Summary: The occurrence of C. neoformans was studied in 261 aviaries at the Antwerp Zoological Gardens during 1985. Droppings of parrots and budgerigars were the most important and constant source of this pathogenic yeast. C. neoformans was also isolated from wooden nesting-boxes.
Zusammenfassung: Das Vorkommen von C. neoformans wurde in 261 Volieren des Zoologischen Gartens zu Antwerpen während des Jahres 1985 untersucht. Exkremente von Papageien und Wellensichttichen waren die wichtigste und konstanteste Quelle dieser pathogenen Hefe. C. neoformans wurde auch aus Holznistkästen isoliert.     

Localized cutaneous cryptococcosis successfully treated with itraconazole. Review of medication in 18 cases reported in Japan

A case of localized cutaneous cryptococcosis successfully treated with oral itraconazole is reported. The patient, a 63-year-old Japanese housewife, had had an erythematous and partly eroded lesion in the area of her right earlobe for about 10 months. No findings suggesting a primary focus of Cryptococcus infection were found in the lung, central nervous system or other internal organs. The isolate obtained from the lesion was identified as Cryptococcus neoformans based on the demonstration of encapsulated blastospores and its biological characteristics. The clinical course shifted toward healing in parallel with a decreased serum level of anticryptococcal antibody after initiation of treatment with oral itraconazole in a dose of 100 mg once daily. The treatment was discontinued after 13 weeks. There were neither subjective nor objective side effects. Furthermore, this paper reviews the medications employed in 18 cases with cutaneous cryptococcosis reported in Japan during the nearly 40-year period between 1952 and 1989.     

Isolation of Cryptococcus neoformans from pigeon (Columba livia) droppings in Mexico City

To determine the ecological and epidemiological significance of pigeon excrement in cryptococcosis in Mexico City, 251 samples of pigeon droppings were studied. These were collected from houses, public buildings, churches, parks and pigeon nests. Each sample was suspended 1 : 10 in isotonic saline solution and then cultured in Staib medium. Identification of Cryptococcus neoformans was performed based on the development of brown-coloured colonies and the presence of encapsulated yeasts. Of 251 samples, 52 (20.7%) were positive for Cr. neoformans. The highest frequency was observed in droppings from public buildings (31.2%), followed by churches (22.0%) and houses (13.3%). No significant differences in isolation frequency were observed between fresh or dried excrement. All isolates obtained were Cr. neoformans var. neoformans. As in other studies on Cr. neoformans from pigeons, performed in other countries, these frequency data are considered ‘normal’. The results, however, accentuate the potential risk of cryptococcosis acquisition, especially now that cryptococcosis frequency is increasing in Mexico, mainly in AIDS patients.     

Cutaneous cryptococcosis as the first symptom of a disseminated cryptococcosis in a patient with lymphogranulomatosis X

A 63-year-old patient, known to have suffered from lymphogranulomatosis X for 4 years is reported, in whom a cutaneous cryptococcosis appeared as first sign of a disseminated cryptococcosis. Despite systemic therapy with amphotericin B, the patient died after a period of 2 months. Differential diagnosis of skin tumours of immunosuppressed patients includes rare skin mycoses, and both histological and mycological examinations should be performed.