Virulence factors, antifungal susceptibility and molecular profile in Candida species isolated from the hands of health professionals before and after cleaning with 70% ethyl alcohol-based gel.

Fungal infections in neonatal intensive care units (NICU) are mainly related to Candida species, with high mortality rates. They are predominantly of endogenous origin, however, cross-infection transmitted by healthcare professionals' hands has occurred. The aim of this study was to identify Candida species isolated from the hands of healthcare professionals in a NICU before and after hygiene with 70% ethanol-based gel and evaluate virulence factors DNase, phospholipase, proteinase, hemolysin, biofilm biomass production, and metabolic activity. In vitro antifungal susceptibility testing and similarity by random amplified polymorphic DNA (RAPD) were also performed. C. parapsilosis complex was the most frequent species (57.1%); all isolates presented at least one virulence factor; three isolates (Candida parapsilosis complex) were resistant to amphotericin B, two (Candida famata [currently Debaryomyces hansenii] and Candida guilliermondii [currently Meyerozyma guilliermondii]) was resistant to micafungin, and six (Candida parapsilosis complex, Candida guilliermondii [=Meyerozyma guilliermondii], Candida viswanathi, Candida catenulata [currently Diutina catenulata] and Candida lusitaniae [currently Clavispora lusitaniae]) were resistant to fluconazole. Molecular analysis by RAPD revealed two clusters of identical strains that were in the hands of distinct professionals. Candida spp. were isolated even after hygiene with 70% ethanol-based gel, highlighting the importance of stricter basic measures for hospital infection control to prevent nosocomial transmission.

Isolation of Cryptococcus species from the external environments of hospital and academic areas.

INTRODUCTION: Fungi of the genus Cryptococcus are cosmopolitan and may be agents of opportunistic mycoses in immunocompromised and sometimes immunocompetent individuals. Cryptococcus species are frequently isolated from trees and bird excreta in the environment and infection occurs by inhalation of propagules dispersed in the air. The aim was to investigate Cryptococcus species in bird excreta and tree hollows located in a university hospital area and in an academic area of a university campus. METHODOLOGY: A total of 40 samples of bird excreta and 41 samples of tree hollows were collected. The identification of the isolates was done by classical methodology and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Twenty (62.5%) isolates of Cryptococcus were found in bird excreta and 12 (37.5%) in tree hollows. C. laurentii (currently Papiliotrema laurentii) was the most frequent species in both samples, being found in 5 samples of excreta and in 8 tree hollows. The diversity of species found in excreta (C. laurentii, C. albidus [currently Naganishia albida], C. liquefaciens [currently N. liquefaciens], C. friedmanii [currently N. friedmannii] and others) was higher than in tree hollows (C. laurentii, C. flavescens [currently Papiliotrema flavescens], and other yeasts). CONCLUSION: Many Cryptococcus species were isolated from excreta and tree hollows, and this fact is important for understanding the environmental epidemiology of those emerging pathogens for public health, as a way to implement surveillance actions and control of cryptococcosis.

The epidemiology of cryptococcosis and the characterization of Cryptococcus neoformans isolated in a Brazilian University Hospital.

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.

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.

Prevalência de sorotipos e resistência antimicrobiana de cepas invasivas do pneumococo em crianças: análise de 9 anos / Prevalence of serotypes and antimicrobial resistance of invasive strains of pneumococcus in children: analysis of 9 years

OBJETIVO: Avaliar o perfil de sorotipos e a sensibilidade aos antimicrobianos de cepas de pneumococo obtidas de crianças e as implicações na formulação de vacinas pneumocócicas. MÉTODOS: Cepas de pneumococo isoladas no Hospital de Clínicas da Universidade Federal de Uberlândia, Uberlândia (MG), a partir de pacientes com doença invasiva, foram enviadas ao Instituto Adolfo Lutz, São Paulo (SP), para confirmação da identificação, sorotipagem e determinação da sensibilidade aos antimicrobianos. RESULTADOS: De abril de 1999 a dezembro de 2008, foram avaliadas 142 cepas de pneumococo obtidas de crianças de até 5 anos de idade. Setenta e cinco (52,8 por cento) eram de pacientes do sexo masculino, e a idade variou de 1 a 60 meses (média de 19±15,4 meses e mediana de 15 meses). Os diagnósticos clínicos mais comuns foram pneumonia [92 casos (64,8 por cento)] e meningite [33 casos (23,2 por cento)], e as principais fontes de recuperação foram sangue [61 amostras (43 por cento)], líquido pleural [52 (36,6 por cento)] e liquor [28 (19,7 por cento)]. Os sorotipos mais comuns foram o 14, 5, 6B, 1, 6A, 18C, 19A, 3, 9V, 19F, 23F, 9N e 10A. Foram detectadas 14 (9,9 por cento) cepas penicilina-resistentes, restritas aos sorotipos 14, 6B, 19F, 19A e 23F e predominantes no período de 2004 a 2008 (p = 0,000). Foi detectada sensibilidade diminuída ao cotrimoxazol (79,5 por cento), à eritromicina e à clindamicina (11,3 por cento cada) e à ceftriaxona (5,6 por cento). CONCLUSÕES: A resistência à penicilina foi detectada em 9,9 por cento das cepas e predominou no período de 2004 a 2008. Foram identificados 20 diferentes sorotipos de pneumococo, e a cifra de cobertura pela vacina 7-valente atualmente disponível (PN CRM7) é de 71,9 por cento.

OBJECTIVE: To determine the prevalence of serotypes and antimicrobial susceptibility of strains of pneumococcus in children and to evaluate the implications for vaccine formulation. METHODS: Strains of pneumococcus obtained from children admitted with invasive diseases were isolated at Hospital de Clínicas of Universidade Federal de Uberlândia, Uberlândia, Brazil, and sent to Instituto Adolfo Lutz, São Paulo, Brazil, for further identification, serotyping, and determination of antimicrobial susceptibility. RESULTS: From April 1999 to December 2008, 142 strains of pneumococcus, obtained from children under 5 years of age, were analyzed. Seventy-five (52.8 percent) patients were male, and the age ranged from 1 to 60 months (mean age = 19±15.4 months; median = 15 months). The most common diagnoses were pneumonia [92 cases (64.8 percent)] and meningitis [33 cases (23.2 percent)]. The strains were mostly isolated from blood [61 samples (43 percent)], pleural fluid [52 samples (36.6 percent)], and cerebrospinal fluid [28 samples (19.7 percent)]. The most common serotypes were 14, 5, 6B, 1, 6A, 18C, 19A, 3, 9V, 19F, 23F, 9N, and 10A. There were 14 [9.9 percent] penicillin-resistant strains, which was detected only in the following serotypes: 14, 6B, 19F, 19A, and 23F, being predominant from 2004 to 2008 (p = 0.000). There was reduced susceptibility to co-trimoxazole (79.5 percent), erythromycin and clindamycin (11.3 percent each), and ceftriaxone (5.6 percent). CONCLUSIONS: Penicillin resistance was detected in 9.9 percent of the strains, being predominant from 2004 to 2008. Twenty different pneumococcal serotypes were identified, and 71.9 percent of the serotypes were represented in the 7-valent conjugate vaccine (PN CRM7) currently available.

New susceptibility breakpoints in antimicrobial resistance rates of invasive pneumococcal strains.

OBJECTIVE: To evaluate the impact of new penicillin susceptibility breakpoints on resistance rates of pneumococcal strains collected from children with pneumonia. METHODS: Pneumococcal strains collected from patients admitted with pneumonia were isolated at the clinical analysis lab of Hospital de Clínicas de Uberlândia, Uberlândia, Brazil, and sent to Instituto Adolfo Lutz, São Paulo, Brazil, for further identification, serotyping and determination of antimicrobial susceptibility. RESULTS: From April 1999 to December 2008, 330 strains of pneumococcus were sent to Instituto Adolfo Lutz; of these, 195 (59%) were collected from patients with pneumonia. One hundred strains collected from patients < or = 12 years old were analyzed. The patients' age ranged from 1 to 12.6 years old (with mean age of 2.4 and median of 1.7 years). Forty-seven patients were male. The strains were isolated from blood (42%) and pleural fluid (58%). There were 35 oxacillin-resistant strains: according to the criteria defined by the Clinical and Laboratory Standards Institute (CLSI) in 2007 [minimum inhibitory concentration (MIC) < or = 0.06 microg/mL for susceptibility (S), 0.12 to 1 microg/mL for intermediate resistance (IR), and > or = 2 microg/mL for full resistance (FR)], 22 strains had IR and 11 strains had FR. According to the current breakpoints defined by the CLSI in 2008 (< or = 2 microg/mL for S, 4 microg/mL for IR and > or = 8 microg/mL for FR), only one strain had IR to penicillin. There was resistance to co-trimoxazole (80%), tetracycline (21%), erythromycin (13%), clindamycin (13%), and ceftriaxone (one strain simultaneously resistant to penicillin). CONCLUSIONS: When the new breakpoints for in vitro susceptibility were applied, penicillin resistance rates dropped 97%, from 33 to 1%.

Prevalence of serotypes and antimicrobial resistance of invasive strains of pneumococcus in children: analysis of 9 years.

OBJECTIVE: To determine the prevalence of serotypes and antimicrobial susceptibility of strains of pneumococcus in children and to evaluate the implications for vaccine formulation. METHODS: Strains of pneumococcus obtained from children admitted with invasive diseases were isolated at Hospital de Clínicas of Universidade Federal de Uberlândia, Uberlândia, Brazil, and sent to Instituto Adolfo Lutz, São Paulo, Brazil, for further identification, serotyping, and determination of antimicrobial susceptibility. RESULTS: From April 1999 to December 2008, 142 strains of pneumococcus, obtained from children under 5 years of age, were analyzed. Seventy-five (52.8%) patients were male, and the age ranged from 1 to 60 months (mean age = 19+/-15.4 months; median = 15 months). The most common diagnoses were pneumonia [92 cases (64.8%)] and meningitis [33 cases (23.2%)]. The strains were mostly isolated from blood [61 samples (43%)], pleural fluid [52 samples (36.6%)], and cerebrospinal fluid [28 samples (19.7%)]. The most common serotypes were 14, 5, 6B, 1, 6A, 18C, 19A, 3, 9V, 19F, 23F, 9N, and 10A. There were 14 [9.9%] penicillin-resistant strains, which was detected only in the following serotypes: 14, 6B, 19F, 19A, and 23F, being predominant from 2004 to 2008 (p = 0.000). There was reduced susceptibility to co-trimoxazole (79.5%), erythromycin and clindamycin (11.3% each), and ceftriaxone (5.6%). CONCLUSIONS: Penicillin resistance was detected in 9.9% of the strains, being predominant from 2004 to 2008. Twenty different pneumococcal serotypes were identified, and 71.9% of the serotypes were represented in the 7-valent conjugate vaccine (PN CRM7) currently available.

Novos pontos de corte de sensibilidade nas taxas de resistência antimicrobiana de cepas invasivas de pneumococo / New susceptibility breakpoints in antimicrobial resistance rates of invasive pneumococcal strains

OBJETIVO: Avaliar impacto dos novos pontos de corte de sensibilidade à penicilina nas taxas de resistência de cepas de pneumococo obtidas de crianças com pneumonia. MÉTODOS: Cepas de pneumococo isoladas no laboratório de análises clínicas do Hospital de Clínicas de Uberlândia, Uberlândia (MG), a partir de amostras de pacientes internados foram enviadas ao Instituto Adolfo Lutz, Sao Paulo (SP), para confirmação da identificação, sorotipagem e determinação da sensibilidade aos antimicrobianos. RESULTADOS: De abril de 1999 a dezembro de 2008 foram enviadas ao Instituto Adolfo Lutz 330 cepas de pneumococo, sendo 195 (59 por cento) provenientes de pacientes com diagnóstico de pneumonia. Destas, foram analisadas 100 cepas de pacientes com idade ≤ 12 anos; a idade dos pacientes variou de 1 a 12,6 anos, com média de 2,4 e mediana de 1,7 anos; 47 pacientes eram do sexo masculino; as fontes de recuperação foram sangue (42 por cento) e líquido pleural (58 por cento). Foram detectadas 35 cepas oxacilina-resistentes: segundo os critérios do Clinical and Laboratory Standards Institute (CLSI) de 2007 [concentração inibitória mínima (CIM) ≤ 0,06 µg/mL para sensibilidade (S), 0,12 a 1 µg/mL para resistência intermediária (RI) e ≥ 2 µg/mL para resistência plena (RP)], 22 cepas apresentaram RI e 11, RP para penicilina. De acordo com os critérios atuais do CLSI de 2008 (≤ 2 µg/mL para S, 4 µg/mL para RI e ≥ 8 µg/mL para RP) apenas uma cepa confirmou RI à penicilina. Detectou-se resistência a cotrimoxazol (80 por cento), tetraciclina (21 por cento), eritromicina (13 por cento), clindamicina (13 por cento) e ceftriaxona (uma cepa, simultaneamente resistente a penicilina). CONCLUSÕES: Com a aplicação dos novos pontos de corte para sensibilidade in vitro, as taxas de resistência a penicilina caíram 97 por cento, de 33 para 1 por cento.

OBJECTIVE: To evaluate the impact of new penicillin susceptibility breakpoints on resistance rates of pneumococcal strains collected from children with pneumonia. METHODS: Pneumococcal strains collected from patients admitted with pneumonia were isolated at the clinical analysis lab of Hospital de Clínicas de Uberlândia, Uberlândia, Brazil, and sent to Instituto Adolfo Lutz, São Paulo, Brazil, for further identification, serotyping and determination of antimicrobial susceptibility. RESULTS: From April 1999 to December 2008, 330 strains of pneumococcus were sent to Instituto Adolfo Lutz; of these, 195 (59 percent) were collected from patients with pneumonia. One hundred strains collected from patients ≤ 12 years old were analyzed. The patients' age ranged from 1 to 12.6 years old (with mean age of 2.4 and median of 1.7 years). Forty-seven patients were male. The strains were isolated from blood (42 percent) and pleural fluid (58 percent). There were 35 oxacillin-resistant strains: according to the criteria defined by the Clinical and Laboratory Standards Institute (CLSI) in 2007 [minimum inhibitory concentration (MIC) ≤ 0.06 µg/mL for susceptibility (S), 0.12 to 1 µg/mL for intermediate resistance (IR), and ≥ 2 µg/mL for full resistance (FR)], 22 strains had IR and 11 strains had FR. According to the current breakpoints defined by the CLSI in 2008 (≤ 2 µg/mL for S, 4 µg/mL for IR and ≥ 8 µg/mL for FR), only one strain had IR to penicillin. There was resistance to co-trimoxazole (80 percent), tetracycline (21 percent), erythromycin (13 percent), clindamycin (13 percent), and ceftriaxone (one strain simultaneously resistant to penicillin). CONCLUSIONS: When the new breakpoints for in vitro susceptibility were applied, penicillin resistance rates dropped 97 percent, from 33 to 1 percent.