Biomedical subjects

C E Canteros

Publications and source records attributed to C E Canteros.

13 recordsLinked to original sources

PubMed1996

A rapid urease test for presumptive identification of Cryptococcus neoformans.

A rapid method to evidence urease activity is described. Urea hydrolysis and consequent production ammonia are detected by a chemical reaction producing a blue phenol compound (indophenol blue). Three hundred and three yeast were tested. Out of 107 urease-positive organisms detected by Christensen's Urea Agar Test (CUAT) 102 were positive by our method. No false negatives were observed by this method when testing 87 Cryptococcus strains. Ths practical screening test for presumptive identification of Cryptococcus neoformans is simple, unaffected by pH changes and requires 15 minutes to be performed.

Clinical Laboratory Techniques↗

PubMedDate not supplied

[In vitro sensitivity of Paracoccidioides brasiliensis to systemically used antifungal agents].

In 1992, the NCCLS proposed a broth macrodilution method for determining the in vitro susceptibility of yeast. However, for dimorphic fungi no standardised method is available. The aim of our study was to evaluate the reference broth macrodilution method with isolates of Paracoccidioides brasiliensis. The in vitro susceptibility of two ATCC strains (32069 and 36324) and nine clinical isolates were determined against serial dilutions of amphotericin B (AMB), ketoconazole (KTZ), and itraconazole (ITZ) from 0.02 to 20 mg/l and fluconazole (FCZ) from 0.1 to 100 mg/l. The inoculum used was 0.5 x 10(4)-2.5 x 10(5) CFU/ml, employing the yeast phase in order to obtain homogeneous growth. The test was incubated with agitation in a rotating shaker at 35 degrees C, and 7 days was the optimal time for MIC final reading. Average MIC50/MIC90 (mg/l) for the eleven isolates tested were the following: AMB: < 0.02/0.16, FCZ: 1.6/6.2, KTZ: 0.08/0.31, ITZ: 0.31/1.3. One clinical isolate showed high MIC for azole drugs (KTZ: 1.3 mg/ml, FCZ: 100 mg/l and ITZ 5 mg/l). Although further studies are necessary to evaluate the usefulness of this methodology for P. brasiliensis, the latter may be clinically useful to predict the development of resistance.

Amphotericin B↗

PubMedDate not supplied

[In vitro infection by different strains of Paracoccidioides brasiliensis].

We analyzed the in vitro infection process by P. brasiliensis and the effect of extracellular factor(s) produced on monolayers of mammalian Vero cell lines. The yeast phase of four strains was studied: B339 (avirulent or slightly virulent), U, (intermediate virulence), 93745 and 63265 (both highly virulent). Strains of intermediate and high virulence had higher adherence at first contact (about 16%). Strain B339 had a slower adherence at first contact (8%) than the others during the same period. The production of extracellular proteases, soluble extracellular factor(s) and extracellular antigen gP43 showed no correlation with the in vitro physiopathogenicity of the analyzed strains. We demonstrate that the Vero model presented in this paper is a suitable system to study infection and virulence in vitro. We are currently assessing its usefulness as a tool for the analysis of the interaction between pathogen, host and antifungal agents.

Animals↗

PubMedDate not supplied

[Distribution of Cryptococcus neoformans serotypes associated with human infections in Argentina].

C. neoformans (Cn) causes severe meningitis in immunocompromised patients, particularly those with AIDS. Little is known about Cn serotypes associated with human infections in Argentina due to the lack of antisera availability. The aim of this study was to produce these antisera, to serotype the Argentinean clinical isolates of Cn received at our laboratory from 1984 to 2001 and to evaluate the agreement of these results with the variety differentiation obtained with Canavanine-Glycine-Bromothymol Blue agar (CGB) medium. We studied 123 isolates recovered from first episodes of cryptococcosis from HIV-infected patients (85), non HIV-infected patients (6) and from other non specified patients (32). Of the isolates, 89% (110/123) were serotype A, 4% (5/123) corresponded to serotype D, 3% (3/129) were serotype AD, 3 isolates were untypable and 2 corresponded to serotype B. All the serotyped isolates agreed with the variety determined by CGB medium. These results indicate that, in our country, most cryptococcal infections in patients with AIDS are associated with serotype A (77/85), which agrees with international studies. In patients with other immunosuppressions, this serotype may be also predominant, although a low number of isolates were tested (4/6). Recovery of Cn serotype B in these clinical isolates suggests that studying the variety and their serotypes might be important to detect a probable epidemiological alteration.

AIDS-Related Opportunistic Infections↗

PubMedDate not supplied

[1st environmental isolation of Cryptococcus neoformans var. gattii in Argentina].

Two strains of C. neoformans var. gattii serotype B were isolated from a park in Buenos Aires city, Argentina. Samples were collected in spring, rubbing with swabs on the inner hollow of Eucalyptus spp. and other trees, which were not identified at the collection moment. Samples were swabbed directly onto cafeic acid with antibiotic agar, then incubated at 28 degrees C and observed daily for a week. Isolates were recovered from cafeic acid medium showing characteristic colonies of the species that allowed differentiation from other microorganisms of the sample. Identification was performed using carbohydrate fermentation, assimilation of nitrogen and carbon sources, and urease and phenoloxidase detection. Strain variety was determined with canavanine-glicine-bromotimol blue agar medium (CGB). Genotypic characterization using AP-PCR with (GACA)4 aleatory primer showed that both band profiles could be differentiated by only one band. These two strains were isolated from tree species different from Eucalyptus spp. These isolates from natural sources demonstrate that C. neoformans var. gattii efosts in Buenos Aires city environment and that it is not necessarily associated with Eucalyptus spp. The presence of C. neoformansvar. gattii in the environment, detected for the first time in Argentina, may explain its sporadic association with human infections in the local population.

Argentina↗

PubMedDate not supplied

[Malassezia species isolated from skin diseases in a care center in the city of Buenos Aires, Argentina].

The epidemiology of Malassezia genus is poorly understood; geographical and population factors might have influence on the species distribution. The aim of this work was to determine the frequency of Malassezia species involved in skin diseases in a diagnostic center at Buenos Aires City. From 02/01/1999 to 10/31/2001, 456 skin specimens from 447 patients diagnosed as pityriasis versicolor, seborrheic dermatitis, foliculitis and atopic dermatitis, were analyzed by microscopic examination and culture. Malassezia spp was evidenced in 255 skin samples (55.9%) by microscopic analysis but it could be recovered from only 99 (38.8%) specimens. Malassezia species were isolated from 10 (3.9%) scarring lesions that were negative by microscopy. Among 109 isolates, M. sympodialis (58.7%) was the most frequent, followed by M. globosa and M. furfur (18.3% and 9.2%). Unique isolates (0.9%) of M. obtusa, M. slooffiae and M. restricta were recovered. Eleven isolates (10.1%) could not be identified because they had lost viability in the culture. This is the first study on Malassezia species associated to skin diseases in Argentina; further screening including other geographic regions of the country is necessary in order to confirm these results.

Adolescent↗

PubMedDate not supplied

[Immunodiagnosis of endemic mycoses and bronchopulmonary aspergillosis: a multicenter study in Argentina].

In order to contribute to the knowledge of the relative frequency of chronic fungal diseases and assess the performance of diagnostic laboratories in Argentina, a multicenter study was performed with the participation of 25 medical centers located in 12 different provinces and Buenos Aires City. Between 04-01-2000 and 03-30-2001, 965 serum specimens from patients clinically suspected of having histoplasmosis (HP), paracoccidioidomycosis (PCM), coccidioidomycosis (CM) or aspergilosis were analyzed. Agar immunodiffusion tests (IDD) were done locally. All positive and 35% of negative sera were retested in the reference center. Results of laboratories of origin showed 98.8% concordance with those of reference center. Antibodies against any of the etiological agents were detected in 120 specimens from 98 patients. Endemic mycoses (HP, PCM and CM) were diagnosed in 70 patients (71.4%) and aspergilosis in 28 (28.6%). The frequencies of the different mycoses in decreasing order were PCM 47 patients (47.9%), aspergilosis 28 patients (28.6%), HP 13 patients (13.3%) and CM 10 patients (10.2%). The study was carried out on a voluntary basis and some areas of the country were not represented. However, the frequencies were in range with the expected rates in the population under study.

Argentina↗

PubMedDate not supplied

[First isolation of Histoplasma capsulatum from the urban bat Eumops bonariensis].

We report the first isolation of Histoplasma capsulatum var. capsulatum from a male bat Eumops bonariensis captured in Buenos Aires city in 2003. The pathogen was recovered from spleen and liver specimens, and was identified by its phenotypic characteristics. PCR with primers 1283, (GTG)5, (GACA)4 and M13 was used to compare both bat isolates with 17 human isolates, 12 from patients residing in Buenos Aires city, and 5 from other countries of the Americas. The profiles obtained with the four primers showed that both bat isolates were identical to each other and closer to Buenos Aires patients than to the other isolates (similarity percentage: 91-100% and 55-97%, respectively). The high genetic relationship between bat isolates and those from patients living in Buenos Aires suggests a common source of infection. This is the first record of E. bonariensis infected with H. capsulatum in the world, and the first isolation of the fungus in the Argentinean Chiroptera population. In the same way as these wild mammals act as reservoir and spread the fungus in the natural environment, infection in urban bats could well be associated with the increase in histoplasmosis clinical cases among immunosuppressed hosts in Buenos Aires city.

Americas↗

PubMedDate not supplied

[Multicenter study of fungemia due to yeasts in Argentina].

The incidence of candidemia has increased approximately 500% in high-complexity hospitals. A change in the spectrum of Candida infections due to species other than Candida albicans has also been detected. Between April 1999 and April 2000 a multicenter study was performed in order to determine the species distribution associated to candidemias in Argentina and the susceptibility profile of the isolates to the current antifungal drugs. Thirty six institutions have participated. All the 265 yeast strains isolated from blood cultures were identified by morphological, physiological, and biochemical tests. The antifungal susceptibility testing of isolates was performed based on the reference NCCLS procedure. The distribution of species was: Candida albicans (40.75%), Candida parapsilosis (28.67%), Candida tropicalis (15.84%), Candida famata (3.77%), Cryptococcus neoformans (3.77%), Candida glabrata (2.64%), and others (4.53%). Most of the isolates were susceptible to amphotericin B, fluconazole and itraconazole. Mortality associated to the fungemia by yeasts episodes (n=265) was 30%, lower than results previously determined (33-54%). The mortality percentage in patients who received antifungal therapy versus patients without treatment was 26.3% and 47%, respectively.

Amphotericin B↗

PubMedDate not supplied

[Histoplasma capsulatum with aberrant morphology isolated in Argentina].

For the first time in Argentina, we describe a strain of Histoplasma capsulatum var. capsulatum with an aberrant morphology that was isolated from a single patient with AIDS. Mycelial phase cultures on agar Sabouraud at 25-28 degrees C showed white, glabrous, umbilicated and centrally radiated colonies. Unusual microscopic findings were the absence of typical conidia, the presence of terminal/intercalary chlamydoconidia with a diameter of 4 pm and of thickened hyphae. Fungal identification was confirmed by the detection of bands H and M species specific antigens in mycelial culture supernatants and reversion to the typical yeast phase on agar brain-heart-cysteine at 37 degrees C. The genomic DNA profile obtained by RAPD-PCR with primers 1281-1283 coincided with the predominant profile of H. capsulatum among isolates from Argentine patients.

AIDS-Related Opportunistic Infections↗

PubMedDate not supplied

[Disk diffusion method for fluconazole susceptibility testing of Candida spp. isolates].

In order to standardize and evaluate a disk diffusion method with visual reading to detect in vitro fluconazole susceptibility of yeast, 1193 clinical isolates were tested. These included 584 Candida albicans, 196 Candida parapsilosis, 200 Candida tropicalis, 113 Candida glabrata, 50 Candida krusei and 50 Candida spp. and other opportunistic yeasts. The disks were manufactured in the INEI-ANLIS "Dr. Carlos G. Malbrán. The disk diffusion method results were compared to MIC results obtained by the reference CLSI M27-A2 broth microdilution method modified by EUCAST. The interpretative breakpoints for in vitro susceptibility testing of fluconazole were established at: zone diameter > or =16 mm for MIC < or =8 microg/ml (susceptible isolates), between 9 and 15 mm for MIC = 16-32 microg/ml (susceptible dose-dependent isolates), and < or =8 mm for MIC > or =64 microg/ml (resistant isolates). Overall agreement between the two methods was 94.7%, with 0.2% very major errors, and 0.3% major errors. Inter - and intralaboratory agreement was good. The disk diffusion method for drug susceptibility testing of Candida spp. isolates is inexpensive, reliable and reproducible. However, when the inhibition zone diameter is < or =15 mm, it is advisable to test the isolate by the reference microdilution method.

Antifungal Agents↗

PubMedDate not supplied

[Incidence of various etiologic agents of superficial mycosis].

In a retrospective review of laboratory records at the Department of Mycology, National Institute of Microbiology "Dr. Carlos G. Malbrán", during the period June 1989-July 1991, 1225 putatively immunocompetent cases of superficial mycoses were identified. Ninety five percent of these patients were adults and 5% children. Among the total cases, dermatomycoses were caused, 67.6% by dermatophytes, 25.9% by yeasts, 5.9% by Malassezia furfur. and 0.5% by other fungi, as proven by the isolation of the etiological agents. (Figure 1, Table 2). Among the dermatophytes Trichophyton rubrum and Trichophyton mentagrophytes greatly predominated accounting for 66.6% and 20.0% of the isolates, respectively, whereas Microsporum canis (8.0%). Epidermophyton floccosum (5.1%) and Microsporum gypseum (0.3%) were found with less frequency (Figure 2). Nails (47%) were the most common source of isolates in adults, followed by feet (28%), smooth skin (15%), groin (5%) and hands (2%) (Table 1). Regarding the relative efficiency of the diagnostic methods, the analyses of laboratory results evidenced that, 98% of the cases with clinical findings compatible with mycoses and 76% of the cases with positive cultures (Table 1) were identified by microscopic observation.

Adult↗

PubMedDate not supplied

[In vitro sensitivity tests for yeasts: evaluation of a micromethod].

A microdilution antifungal susceptibility test for yeasts was evaluated, based on the macro broth dilution method standardized by the National Committee for Clinical Laboratory Standards (NCCLS) Subcommittee on Antifungal Susceptibility Testing. Both methods were compared using six reference strains with different patterns of susceptibility to the following antifungal drugs: amphotericin B (AMB), flucytosine (5FC), fluconazole (FCZ), itraconazole (ITZ), ketoconazole (KTZ) and miconazole (MCZ). Minimal inhibitory concentration (MIC) results obtained by both methods differed only in 1 or 2 dilutions. Microdilution MIC's determined as visual endpoints and as quantitative measurement of 80% inhibition of relative growth showed a significant correlation (p < 0.001) for AMB, 5FC, FCZ, MCZ and ITZ, conversely no correlation (p = 1.00) for KTZ was observed as determined with 47 local isolates of Candida albicans.

Antifungal Agents↗