Fc receptor-like 3 (-169T>C) polymorphism increases the risk of tendinopathy in volleyball athletes: a case control study.

BACKGROUND: Tendinopathy pathogenesis is associated with inflammation. Regulatory T (Treg) cells contribute to early tissue repair through an anti-inflammatory action, with the forkhead box P3 (FOXP3) transcription factor being essential for Treg function, and the FC-receptor-like 3 (FCRL3) possibly negatively regulating Treg function. FCRL3 -169T>C and FOXP3 -2383C>T polymorphisms are located near elements that regulate respective genes expression, thus it was deemed relevant to evaluate these polymorphisms as risk factors for tendinopathy development in athletes. METHODS: This case-control study included 271 volleyball athletes (146 tendinopathy cases and 125 controls) recruited from the Brazilian Volleyball Federation. Genotyping analyses were performed using TaqMan assays, and the association of the polymorphisms with tendinopathy evaluated by multivariate logistic regression. RESULTS: Tendinopathy frequency was 63% patellar, 22% rotator cuff and 15% Achilles tendons respectively. Tendinopathy was more common in men (OR = 2.87; 95% CI = 1.67-4.93). Higher age (OR = 8.75; 95% CI = 4.33-17.69) and more years of volleyball practice (OR = 8.38; 95% CI = 3.56-19.73) were risk factors for tendinopathy. The FCRL3 -169T>C frequency was significantly different between cases and controls. After adjustment for potential confounding factors, the FCRL3 -169C polymorphism was associated with increased tendinopathy risk (OR = 1.44; 95% CI = 1.02-2.04), either considering athletes playing with tendon pain (OR = 1.98; 95% CI = 1.30-3.01) or unable to train due to pain (OR = 1.89; 95% CI = 1.01-3.53). The combined variant genotypes, FCRL3 -169TC or -169CC and FOXP3 -2383CT or -2383TT, were associated with an increased risk of tendinopathy among athletes with tendon pain (OR = 2.24; 95% CI: 1.14-4.40 and OR = 2.60; 95% CI: 1.11-6.10). The combined analysis of FCRL3 -169T>C and FOXP3 -2383C>T suggests a gene-gene interaction in the susceptibility to tendinopathy. CONCLUSIONS: FCRL3 -169C allele may increase the risk of developing tendinopathy, and together with knowledge of potential risk factors (age, gender and years playing) could be used to personalize elite athletes' training or treatment in combination with other approaches, with the aim of minimizing pathology development risk.

Intestinal Lesion in a Dog Due to Cryptococcus gattii Type VGII and Review of Published Cases of Canine Gastrointestinal Cryptococcosis.

Cryptococcosis is a mycosis caused by yeasts of genus Cryptococcus, mainly the species C. neoformans and C. gattii that can affect humans and animals. These yeasts are widely distributed in the environment and are typically associated with avian droppings and decaying wood. Most infections are related to the respiratory tract, but the central nervous system and cutaneous lesions are also reported in the literature. The present report is a case of cryptococcosis in an 18-month-old unspayed female English Bulldog with the main complaint of weight loss and diarrhea. The presence of two large masses observed in an ultrasound examination leads us to perform an exploratory laparotomy. Considering the size of the lesion and the impossibility of owner to provide intensive care, the consent for euthanasia was requested. The postmortem diagnosis of cryptococcosis was revealed by cytological evaluation, and the involvement of C. gattii VGII was confirmed by isolation and identification tests as well as by the detection of the URA5 gene restriction fragment length polymorphism PCR analysis. Reports in the literature of the involvement of Cryptococcus in gastrointestinal lesions are rare in both human and veterinary medicine. Data about different forms of cryptococcosis are important to provide more knowledge of uncommon clinical presentations of this yeast and therefore improve the diagnoses and decisions for the best therapy.

Asymptomatic cryptococcal antigen prevalence detected by lateral flow assay in hospitalised HIV-infected patients in São Paulo, Brazil.

OBJECTIVE: To determine the prevalence of asymptomatic cryptococcal antigen (CRAG) using lateral flow assay (LFA) in hospitalised HIV-infected patients with CD4 counts <200 cells/µl. METHODS: Hospitalised HIV-infected patients were prospectively recruited at Instituto de Infectologia Emilio Ribas, a tertiary referral hospital to HIV-infected patients serving the São Paulo State, Brazil. All patients were >18 years old without prior cryptococcal meningitis, without clinical suspicion of cryptococcal meningitis, regardless of antiretroviral (ART) status, and with CD4 counts <200 cells/µl. Serum CRAG was tested by LFA in all patients, and whole blood CRAG was tested by LFA in positive cases. RESULTS: We enrolled 163 participants of whom 61% were men. The duration of HIV diagnosis was a median of 8 (range, 1-29) years. 26% were antiretroviral (ART)-naïve, and 74% were ART-experienced. The median CD4 cell count was 25 (range, 1-192) cells/µl. Five patients (3.1%; 95%CI, 1.0-7.0%) were asymptomatic CRAG-positive. Positive results cases were cross-verified by performing LFA in whole blood. CONCLUSIONS: 3.1% of HIV-infected inpatients with CD4 <200 cells/µl without symptomatic meningitis had cryptococcal antigenemia in São Paulo, suggesting that routine CRAG screening may be beneficial in similar settings in South America. Our study reveals another targeted population for CRAG screening: hospitalised HIV-infected patients with CD4 <200 cells/µl, regardless of ART status. Whole blood CRAG LFA screening seems to be a simple strategy to prevention of symptomatic meningitis.

Cryptococcosis in non-HIV/non-transplant patients: A Brazilian case series.

Cryptococcosis is a classical systemic opportunistic mycosis, primarily occurring among patients with significant immunologic impairment. However, this disease could also affect patients without any recognized immunologic defects, that is, phenotypically normal patients. The medical records of 29 non-HIV/nontransplant patients with cryptococcal disease during the period 2007-2014 were retrospectively reviewed. The most common site of infection was the central nervous system (n = 25, 86.2%), followed by the pulmonary system (n = 11, 37.9%) and blood (n = 2, 6.8%). Thoracic- and brain-computed tomography demonstrated abnormalities of 81.2% (n = 13) and 62.5% (n = 15), respectively. In sum, 22% (n = 6) of the patients experienced a significant underlying condition. More than one therapeutic regimen was used in 77.8% (n = 21) of the patients. The isolates were identified as being Cryptococcus neoformans species complex (n = 4, 36.4%) and Cryptococcus gattii species complex (n = 7, 63.6%). The overall mortality was 20.7% (n = 6). Herein, we presented the first case series of cryptococcosis in this specific population in São Paulo City, Brazil. The incidence of cryptococcosis in our hospital has not increased in recent years, and 77.8% (n = 21) of cases had no obvious predisposing factor. However, this disease remains associated with high mortality.

Feline nasal granuloma due to Cryptoccocus gattii type VGII.

The aims of this study are to make a more precise identification of the etiologic agent of a nasal granuloma in a cat, to verify the susceptibility to the antifungal drugs: ketoconazole, itraconazole, fluconazole, posaconazole, voriconazole, amphotericin B and the proper treatment. Part of the granuloma's fragment was removed, added to a saline solution and sent to the Laboratory of Mycology. The solution was then seeded in Sabouraud dextrose agar, and the yeast was primarily identified by the traditional methods. The confirmation of the specie Cryptococcus gattii and its molecular type were performed using the PCR-RFLP molecular techniques. The antifungal susceptibility was verified by using the E-test method, and the cat was treated with itraconazole associated with 5-flucytosine. The isolated strain was identified as C. gattii type VGII and was susceptible to all antifungal drugs tested. The treatment with itraconazole associated with 5-flucytosine led to the cure of granulomatous lesions in the feline after 6 months. The characterization and molecular investigation of this microorganism are relevant because they could help us better understand the epidemiology of the infection and to guide us to treat properly the disease.

Disseminated amphotericin-resistant fusariosis in acute leukemia patients: report of two cases.

Disseminated fusariosis has emerged as a significant, usually fatal infection in immunocompromised hosts despite antifungal treatment. We describe here two patients with acute leukemia who developed disseminated amphotericin-resistant fusariosis, and review of six studies of cases series in the literature. Two Fusarium solani strains were isolated from blood and skin cultures of one patient, and one strain from the blood culture of the second patient. Both patients died despite antifungal treatment. Strains were identified by sequencing of ITS1 and ITS4 regions. Random amplified polymorphic DNA analysis of the three F. solani isolates showed a low degree of similarity. Screening for Fusarium spp. contaminants within our facility was negative. Using the CLSI M-38-A2 broth dilution method and E tests(®), we found that the MICs were low for voriconazole (0.12 and 0.5 mg/L, respectively), unexpectedly high for amphotericin B (≥8 and ≥32 µg/mL, respectively) and itraconazole (≥16 mg/ml). Patients with leukemia or persistent neutropenia should be assessed for disseminated fungal infections, including biopsy and skin cultures. Antifungal susceptibility tests are important due to the possibility of the strains being amphotericin resistant. Treatments must be aggressive, with high doses of antifungals or combined therapy.

Comparison of the broth microdilution (BMD) method of the European Committee on Antimicrobial Susceptibility Testing and the Clinical Laboratory Standards Institute BMD method for non-Candida albicans and non-C. tropicalis bloodstream isolates from eleven tertiary hospitals in São Paulo state, Brazil.

We aim in this study to provide levels of susceptibility of 162 bloodstream isolates of non-Candida albicans and non-C. tropicalis species from a sentinel program conducted in 11 hospitals in Brazil. Additionally, we compared the broth microdilution (BMD) method of the European Committee of Susceptibility Testing (EUCAST) with Clinical Laboratory Standards Institute (CLSI) BMD method for fluconazole, itraconazole, voriconazole, and amphotericin B. The study included 103 C. parapsilosis, 38 C. glabrata, 8 C. orthopsilosis, and 7 C. krusei isolates, and single isolates of Pichia anomala, C. famata, C. lusitaniae, C. kefyr, C. guilliermondii, and C. metapsilosis. Of note, we observed cross-resistance between fluconazole and voriconazole for two isolates being one C. parapsilosis and one C. glabrata. Good essential agreement (EA) was observed between the EUCAST and the CLSI results for C. parapsilosis and for fluconazole, itraconazole, voriconazole, and amphotericin B, respectively: 98%, 99%, 98%, and 97%. Otherwise, for C. glabrata, the EA for fluconazole was 84.2% and for voriconazole 89.4%. Because data from Brazil are scarce, our results contribute to the consolidation of the database of candidemia agents and monitoring of trends in the profile of drug resistance.

Cerebral infection caused by Cryptococcus gattii: a case report and antifungal susceptibility testing.

We report a clinical case of cerebral infection caused by Cryptococcus gattii in a 10 year-old boy. Clinical and laboratory exams did not demonstrate any apparent immunosuppressed state (HIV antibody and the tuberculin skin tests, both negative, were performed; blood cells count and immunoglobulin levels were within normality). Treatment was begun with amphotericin B-deoxycholate but renal toxicity signs led to its substitution by fluconazole. The infection proceeded even after treatment with fluconazole. In vitro determination of minimum inhibitory concentration values were high for itraconazole (= or > 2 microg/ml), fluconazole and 5-flucytosine (= or > 64 microg/ml) and low for amphotericin B (1.0 microg/ml). Renal toxicity signs, induced by amphotericin B, progression of infection after fluconazole, and likely in vivo resistance to this triazole made this case difficult to treat. In vitro drug interaction tests confirmed probable synergism between amphotericin B and 5-flucytosine (frational inhibitory concentration - FIC = 0.375). In contrast, a probable additive effect was observed for amphotericin B and fluconazole (FIC = 0.75). Initial treatment of persistent high intracranial pressure was insufficient and neurological surgery was necessary. Antifungal susceptibility tests and Cryptococcus species identification were important in selecting appropriate antifungal therapy.

Comparative analysis of quadriceps and hamstrings strength in knee osteoarthritis before and after total knee arthroplasty: a cross-sectional study.

OBJECTIVE: Compare the maximal isokinetic muscle strength of knee extensor and flexor muscles between patients with knee osteoarthritis and patients submitted to total knee arthroplasty. METHODS: Volunteers were divided into five groups (n = 20): Control; Ahlbäck I and II; Ahlbäck IV; six months after total knee arthroplasty; 12 months after total knee arthroplasty. An isokinetic knee strength evaluation was conducted for the quadriceps and hamstrings at 60°/s. RESULTS: Significant differences in the peak torque of the quadriceps and hamstrings were found among the groups (p < 0.001). The Ahlbäck IV, six-month, and 12-month postoperative groups demonstrated lower values when compared to the Control and Ahlbäck I and II groups. When percentage values were compared to the Control group, mean differences ranged from 7% to 41%. CONCLUSION: Patients with healthy knees or early stage osteoarthritis have higher quadriceps and hamstrings strengths than those with a more advanced stage of the disease, even after knee replacement. These findings suggest that the traditional rehabilitation programs do not recover strength to levels observed in individuals without knee osteoarthritis.

OBJETIVO: Comparar a força muscular isocinética máxima dos músculos extensores e flexores do joelho entre pacientes com osteoartrite do joelho e pacientes submetidos à artroplastia total do joelho. MÉTODOS: Os voluntários foram divididos em cinco grupos (n = 20): Controle, Ahlbäck I e II; Ahlbäck IV; seis meses após artroplastia total do joelho; 12 meses após artroplastia total do joelho. O teste de força voluntária isocinética máxima foi feito para mensuração da força do quadríceps e isquiotibiais a 60/s. RESULTADOS: Foram achadas diferenças significativas entre o pico de torque do quadríceps e dos isquiotibiais (p < 0,001). Os grupos Ahlbäck IV, seis meses e 12 meses após cirurgia mostraram valores mais baixos quando comparados com os grupos controle e Ahlbäck I e II. Quando os valores percentuais foram comparados com o grupo Controle, as diferenças médias variaram de 7% a 41%. CONCLUSÃO: Os pacientes com joelhos saudáveis ou osteoartrite em estágio inicial apresentaram maior força no quadríceps e nos isquiotibiais do que pacientes em estágio mais avançado da doença, mesmo após a ATJ. Esses achados sugerem que os programas tradicionais de reabilitação não recuperam a força nos níveis observados em indivíduos sem osteoartrite do joelho.

Antifungal Susceptibility of Candida Species Isolated from Horticulturists with Onychomycosis in Piauí, Brazil.

BACKGROUND: We aimed to assess susceptibility pattern of Candida species isolated from horticulturists with onychomycosis to four antifungal drugs and to compare the effectiveness of conventional identification methods with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS). METHODS: This study was conducted in a community garden located in Teresina, State of Piauí, Brazil, in the year 2014. The samples were identified through phenotypic methods and per MALDI-TOF MS, being used PCR as definitive identification test. The susceptibility pattern to four antifungal drugs was determined according to Clinical and Laboratory Standards Institute (CLSI). RESULTS: Fourteen clinical isolates from seven different species were identified by the phenotypic method and by MALDI-TOF MS, with an observed concordance of 71.4% between the two methods. C. albicans (28.6%), C. parapsilosis (21.4%), C. guilliermondii and C. metapsilosis (both with 14.3%) were the most frequent species. With the exception of C. krusei, all species were sensitive to the tested antifungal. CONCLUSION: This is the first study of antifungal susceptibility of Candida in Piauí, Brazil. With the exception of C. krusei, no species showed resistance to the antifungal drugs used. This study suggests constants updates from the public databases used in MALDI-TOF MS to provide a rapid and accurate mycological diagnosis.

Performance of cryptococcal antigen lateral flow assay in serum, cerebrospinal fluid, whole blood, and urine in HIV-infected patients with culture-proven cryptococcal meningitis admitted at a Brazilian referral center.

Cryptococcal meningitis is the most common cause of opportunistic meningitis in HIV-infected patients in Brazil and causes unacceptable high mortality rates. In this study, HIV-infected patients with a first episode of culture-proven cryptococcal meningitis in cerebrospinal fluid (CSF) were prospectively included in order to evaluate sensitivity of cryptococcal antigen (CrAg) lateral flow assay (LFA) in serum, CSF, whole blood (fingerstick), and fresh urine. In addition, HIV-infected patients with other neurological confirmed diseases were included in order to evaluate the specificity of CrAg LFA in serum. Twenty patients with cryptococcal meningitis were included and in 19 of them, CrAg LFA in CSF, serum, and whole blood were positive (95% sensitivity). In 18 patients, India ink test was positive in CSF (90% sensitivity), and in 16 cases, CrAg LFA was positive in urine (80% sensitivity). Thirty-six HIV-infected patients with other neurological diseases had negative results of CrAg LFA in serum (100% specificity). In conclusion, CrAg LFA in serum, CSF, and whole blood showed high sensitivity and specificity. Whole blood CrAg LFA seems to be a good and reliable strategy to improve AIDS-related cryptococcal meningitis diagnosis in Brazil.

Candidaemia due to Candida parapsilosis species complex at a hospital in Brazil: Clinical characteristics and antifungal susceptibility profile.

BACKGROUND: Recent decades have seen a global emergence of candidaemia caused by non-Candida albicans Candida species, particularly the Candida parapsilosis complex. AIMS: To evaluate the clinical features and antifungal susceptibility profiles of isolates belonging to the C. parapsilosis species complex in patients with candidaemia in a midwestern Brazilian tertiary-care teaching hospital. METHODS: Yeast identification was performed using an automated Vitek 2 Compact system. PCR-RFLP was employed for species differentiation. RESULTS: Five cases of infection by C. parapsilosis sensu stricto and two by Candida orthopsilosis were found. Of the seven cases, five were adult patients undergoing haemodialysis. The only isolate of C. parapsilosis sensu stricto resistant to fluconazole (MIC=8µg/ml) was obtained from a patient on a long-term regimen with this drug. This was the only patient who evolved to death. CONCLUSIONS: Resistance to antifungal agents poses a therapeutic challenge, especially for non-C. albicans Candida species, and requires continuous monitoring using susceptibility tests because resistance in vitro can be predictive of treatment failure. In the present study, in vitro antifungal susceptibility proved consistent with clinical outcome.

Population Genetic Analysis Reveals a High Genetic Diversity in the Brazilian Cryptococcus gattii VGII Population and Shifts the Global Origin from the Amazon Rainforest to the Semi-arid Desert in the Northeast of Brazil.

Cryptococcus neoformans and Cryptococcus gattii are responsible globally for almost one million cryptococcosis cases yearly, mostly in immunocompromised patients, such as those living with HIV. Infections due to C. gattii have mainly been described in tropical and subtropical regions, but its adaptation to temperate regions was crucial in the species evolution and highlighted the importance of this pathogenic yeast in the context of disease. Cryptococcus gattii molecular type VGII has come to the forefront in connection with an on-going emergence in the Pacific North West of North America. Taking into account that previous work pointed towards South America as an origin of this species, the present work aimed to assess the genetic diversity within the Brazilian C. gattii VGII population in order to gain new insights into its origin and global dispersal from the South American continent using the ISHAM consensus MLST typing scheme. Our results corroborate the finding that the Brazilian C. gattii VGII population is highly diverse. The diversity is likely due to recombination generated from sexual reproduction, as evidenced by the presence of both mating types in clinical and environmental samples. The data presented herein strongly supports the emergence of highly virulent strains from ancestors in the Northern regions of Brazil, Amazonia and the Northeast. Numerous genotypes represent a link between Brazil and other parts of the world reinforcing South America as the most likely origin of the C. gattii VGII subtypes and their subsequent global spread, including their dispersal into North America, where they caused a major emergence.

Report of filamentous forms in a mating type VNI clinical sequential isolates of Cryptococcus neoformans from an HIV virus-infected patient.

We reported a cryptococcal meningitis Aids-patient infected with a mating type VNI isolate showing filamentous cells in direct examination of cerebrospinal fluid. Clinical data, outcome, treatment features and microbiological findings were discussed.

Molecular diagnosis of cryptococcal meningitis in cerebrospinal fluid: comparison of primer sets for Cryptococcus neoformans and Cryptococcus gattii species complex.

AIM: This study evaluated the use of polymerase chain reaction for cryptococcal meningitis diagnosis in clinical samples. MATERIALS AND METHODS: The sensitivity and specificity of the methodology were evaluated using eight Cryptococcus neoformans/C. gattii species complex reference strains and 165 cerebrospinal fluid samples from patients with neurological diseases divided into two groups: 96 patients with cryptococcal meningitis and AIDS; and 69 patients with other neurological opportunistic diseases (CRL/AIDS). Two primer sets were tested (CN4-CN5 and the multiplex CNa70S-CNa70A/CNb49S-CNb-49A that amplify a specific product for C. neoformans and another for C. gattii). RESULTS: CN4-CN5 primer set was positive in all Cryptococcus standard strains and in 94.8% in DNA samples from cryptococcal meningitis and AIDS group. With the multiplex, no 448-bp product of C. gattii was observed in the clinical samples of either group. The 695bp products of C. neoformans were observed only in 64.6% of the cryptococcal meningitis and AIDS group. This primer set was negative for two standard strains. The specificity based on the negative samples from the CTL/AIDS group was 98.5% in both primer sets. CONCLUSIONS: These data suggest that the CN4/CN5 primer set was highly sensitive for the identification of C. neoformans/C. gattii species complex in cerebrospinal fluid samples from patients with clinical suspicion of cryptococcal meningitis.

Primary cutaneous cryptococcosis due to Cryptococcus neoformans var. gattii serotype B, in an immunocompetent patient.

The authors report a male patient, a seller with no detected immunosuppression, with an extensive ulcerated skin lesion localized on the left forearm, caused by Cryptococcus neoformans var. gattii serotype B. Oral treatment with fluconazole was successful. A review of the literature showed the rarity of this localization in HIV-negative patients. In contrast, skin lesions frequently occurs in HIV-positive patients, with Cryptococcus neoformans var. neoformans serotype A predominating as the etiological agent. In this paper, the pathogenicity of C. neoformans to skin lesions in patients immunocompromised or not, is discussed, showing the efficacy of fluconazole for the treatment of these processes.

Comparative analysis of quadriceps and hamstrings strength in knee osteoarthritis before and after total knee arthroplasty: a cross-sectional study / Análise comparativa da força do quadríceps e dos isquiotibiais na osteoartrite do joelho antes e após a artroplastia total do joelho: um estudo transversal

ABSTRACT Objective: Compare the maximal isokinetic muscle strength of knee extensor and flexor muscles between patients with knee osteoarthritis and patients submitted to total knee arthroplasty. Methods: Volunteers were divided into five groups (n = 20): Control; Ahlbäck I and II; Ahlbäck IV; six months after total knee arthroplasty; 12 months after total knee arthroplasty. An isokinetic knee strength evaluation was conducted for the quadriceps and hamstrings at 60°/s. Results: Significant differences in the peak torque of the quadriceps and hamstrings were found among the groups (p < 0.001). The Ahlbäck IV, six-month, and 12-month postoperative groups demonstrated lower values when compared to the Control and Ahlbäck I and II groups. When percentage values were compared to the Control group, mean differences ranged from 7% to 41%. Conclusion: Patients with healthy knees or early stage osteoarthritis have higher quadriceps and hamstrings strengths than those with a more advanced stage of the disease, even after knee replacement. These findings suggest that the traditional rehabilitation programs do not recover strength to levels observed in individuals without knee osteoarthritis.

RESUMO Objetivo: Comparar a força muscular isocinética máxima dos músculos extensores e flexores do joelho entre pacientes com osteoartrite do joelho e pacientes submetidos à artroplastia total do joelho. Métodos: Os voluntários foram divididos em cinco grupos (n = 20): Controle, Ahlbäck I e II; Ahlbäck IV; seis meses após artroplastia total do joelho; 12 meses após artroplastia total do joelho. O teste de força voluntária isocinética máxima foi feito para mensuração da força do quadríceps e isquiotibiais a 60/s. Resultados: Foram achadas diferenças significativas entre o pico de torque do quadríceps e dos isquiotibiais (p < 0,001). Os grupos Ahlbäck IV, seis meses e 12 meses após cirurgia mostraram valores mais baixos quando comparados com os grupos controle e Ahlbäck I e II. Quando os valores percentuais foram comparados com o grupo Controle, as diferenças médias variaram de 7% a 41%. Conclusão: Os pacientes com joelhos saudáveis ou osteoartrite em estágio inicial apresentaram maior força no quadríceps e nos isquiotibiais do que pacientes em estágio mais avançado da doença, mesmo após a ATJ. Esses achados sugerem que os programas tradicionais de reabilitação não recuperam a força nos níveis observados em indivíduos sem osteoartrite do joelho.

Susceptibility to antifungal agents and genotypes of Brazilian clinical and environmental Cryptococcus gattii strains.

There are few reports concerning the in vitro antifungal susceptibility of clinical and environmental Cryptococcus gattii isolates. In this study, we performed polymerase chain reaction-restriction fragment length polymorphism to investigate the molecular subtypes of 50 clinical and 4 environmental Brazilian isolates of C. gattii and assessed their antifungal susceptibility for fluconazole (FLU) and amphotericin B (Amb) according to recent recommendations proposed for antifungal susceptibility testing of nonfermentative yeasts. Time-kill curve studies were performed using RPMI 1640 medium to analyze the fungicidal effect of AmB. We found 47 VGII (94%) molecular types and 3 VGI (6%) types among the clinical isolates. The environmental isolates were VGII (75%) subtype and VGI (25%) subtype. The FLU-MIC ranged from 1 to 64 mg L(-1), and MIC(50)/MIC(90) values were, respectively, 8/16 mg L(-1). For AmB, the MICs were low and homogeneous, ranging from 0.12 to 0.5 mg L(-1), for VGI or VGII. The time required to reach the fungicidal end point (99.9% killing) was 6 h for the majority of strains (64%), but viable cells of VGII were still present after 48 h of exposition. We pointed out the occurrence of high FLU-MICs for C. gattii isolates with highest values for VGII. Our data also suggest that the rate of killing of C. gattii by AmB is strain dependent, and viable cells of VGII genotype strains were still observed after an extended incubation time, addressing future studies to determine whether the in vitro fungicidal activity could be clinically relevant.

Prevalence, distribution and antifungal susceptibility profiles of Candida parapsilosis, Candida orthopsilosis and Candida metapsilosis bloodstream isolates.

The Candida parapsilosis group encompasses three species: C. parapsilosis, Candida orthopsilosis and Candida metapsilosis. These species are phenotypically indistinguishable, and molecular methods are needed for their detection. We analysed 152 unique blood culture isolates of the C. parapsilosis group obtained during 1997-2011. The isolates were screened by PCR amplification of the gene encoding secondary alcohol dehydrogenase, followed by digestion with the restriction enzyme BanI. Isolates with RFLP patterns distinct from those of the C. parapsilosis group were characterized as C. parapsilosis sensu stricto (90.8 %), C. orthopsilosis (8.6 %) and C. metapsilosis (0.6 %). Antifungal susceptibility tests indicated that all isolates were susceptible to itraconazole, amphotericin B and caspofungin. Although C. orthopsilosis and C. metapsilosis isolates were susceptible to fluconazole, higher MICs (≥2 mg l(-1)) were observed for C. orthopsilosis. Three isolates (2.0 %) of C. parapsilosis sensu stricto were resistant to voriconazole. Five C. parapsilosis isolates (3.3 %) were intermediate, and a single isolate (0.7 %) was resistant (MIC 16 mg l(-1)) to fluconazole. These data were confirmed using reference strains. It was observed that C. parapsilosis isolates were less susceptible to all triazoles, and this finding deserves further attention to assess the appearance of cross-resistance phenomena. In conclusion, C. metapsilosis and C. orthopsilosis are involved in a small but significant number of invasive infections in Brazil.

[Mycological aspects and susceptibility in vitro the yeast of the genus Candida from HIV-positive patients in the State of Mato Grosso]. / Aspectos micológicos e suscetibilidade in vitro de leveduras do gênero Candida em pacientes HIV-positivos provenientes do Estado de Mato Grosso.

INTRODUCTION: Candidiasis is one of the most common fungal infections among patients infected by human immunodeficiency virus. The present study aimed to characterize yeasts of the genus Candida from distinct clinical samples from HIV-positive patients and determine the in vitro susceptibility profile to five antifungal drugs. METHODS: Characterization of Candida sp was achieved using the classic methodology: biochemical (zymogram and auxanogram) and micromorphology (germinative tube growth test and slide microculture) tests. Genotypic technique (PCR) and identification by the commercial method API 20C AUX (Biomeriéux) were also performed. To determine the in vitro susceptibility profile, five antifungal drugs were used (ketoconazole, fluconazole, itraconazole, voriconazole and amphotericin-B) following a commercially available method, the Etest. RESULTS: The procedure isolated 105 yeasts of the genus Candida from 102 HIV-infected patients. Of these, 82 (78.1%) were characterized as Candida albicans, 8 (7.6%) as C. parapsilosi s, 8 (7.6%) C. tropicalis, 4 (3.8%) C. krusei, 2 (1.9%) C. glabrata, and 1 (1%) as C. guiilliermondii. CONCLUSIONS: Considering the general profile of sensitivity, 60% of isolates were susceptible to all the antifungal drugs tested; however, the species C. tropicalis and C. krusei showed a tendency toward higher MICs to azoles than those obtained for C. albicans, suggesting resistance.