agar and Meningitis--Cryptococcal

Cryptococcal meningitis (CM) is a life threatening disease and leading cause of opportunistic fungal-related mortality in HIV/AIDS. Most CM infections are caused by C. neoformans species complexes but the prevalence of Cryptococcus gattii species complexes in Uganda is unknown however, it is known in a few other parts of Africa. We estimated the prevalence of C. gattii in patients living with HIV and a diagnosis of cryptococcal meningitis in Uganda.. Cryptococcus isolates (n = 200) obtained from cerebrospinal fluid of patients with CM recruited at the Infectious Diseases Institute, Kampala, Uganda, were tested by phenotypic methods. The Cryptococcus isolates were sub-cultured on Sabouraud Dextrose Agar plates for 48 hours. The yeast colonies were examined by India ink stain, urea hydrolysis, and C. gattii was identified by blue pigmentation on CGB agar. The results were analyzed for frequency of C. gattii. Patient demographic characteristics were collected from the case record forms.. From the 200 patients' case record forms, 87 (43.5%) were female and 113 (56.5%) were male. The median age was 35 (19-64) years. Most patients, 93% (187/200) were from Central Uganda in the districts of Kampala and Wakiso. 97.51% (157/161) of the patients had absolute CD4 lymphocyte counts of less than 200 cells per cubic millimeter; 1.86% (3/161) 200-350 cells per cubic millimeter and 0.62% (1/161) above 500 cells per cubic millimeter. 45.4% (74/163) were not yet on HAART and 54.6% (89/163) were on HAART. 66.7% (58/87) had poor adherence to HAART treatment and 33.3% (29/87) had reported good adherence to HAART treatment. A total of 200 clinical isolates of Cryptococcus isolates were tested. No (0% (0/200) C. gattii was identified among the Cryptococcus isolates.. In this study among patients living with HIV and a diagnosis of cryptococcal meningitis in Uganda, we found no C. gattii infections.

Topics: Acquired Immunodeficiency Syndrome; Adult; Agar; Cryptococcosis; Cryptococcus gattii; Cryptococcus neoformans; Female; Humans; Male; Meningitis, Cryptococcal; Prevalence; Uganda

The Neo-Sensitabs diffusion method was evaluated for determining the antifungal susceptibilities of 30 Cryptococcus gattii isolates to amphotericin B (AMB), fluconazole (FLC), itraconazole (ITC) and voriconazole (VRC). Three different culture media, Müeller-Hinton (MH), RPMI 1640 (RPMI) and Antibiotic medium 3 (AM3), all supplemented with 2% of glucose and 0.5 microg/ml of methylene blue, were tested. The tests were repeated three times on different days at three incubation times (48, 72 and 96 h). Results were compared with those obtained with the CLSI M27-A2 broth microdilution method. The degree of reproducibility of the diffusion test was 100% for VRC and ITC, 98.3-100% for AMB and 43.3-73.3% for FLC. The best reproducibility was observed at 48 h of incubation and no important differences among media were observed at any of the incubation times assayed. Between Neo-Sensitabs and the reference method, VRC showed the best agreement and ITC the worst in all conditions tested (100% and 56.7%, respectively). AMB showed a high agreement between the two methods (93.3% to 96.7%) but Neo-Sensitabs assay failed to detect resistant isolates (discrepancy classified as "very major error") in all times of incubation assayed. Only agreement between both methods for FLC was clearly affected by incubation time and media used, the best results being achieved at 48 h of incubation when MH and RPMI (80.0%, in both media) were used.

Topics: Agar; Amphotericin B; Antifungal Agents; Cryptococcus; Culture Media; Diffusion; Drug Resistance, Fungal; Fluconazole; Humans; Itraconazole; Meningitis, Cryptococcal; Microbial Sensitivity Tests; Pyrimidines; Reproducibility of Results; Triazoles; Voriconazole