Gilselena Kerbauy

Virulence determinants in vancomycin-resistant Enterococcus faecium vanA isolated from different sources at University Hospital of Londrina, Paraná, Brazil

Enterococcus faecium, especially those showing multidrug resistance, has emerged as a significant cause of healthcare-associated infections worldwide. However, relatively little is known about the virulence and pathogenesis of this species. The aim of this study was to determine the occurrence of four putative virulence determinants of E. faecium and to correlate them with phenotypic traits. Using forty E. faecium vanA-type isolates from hospitalized patients and their environmental vicinity, we determined the following: the antimicrobial susceptibility profile, occurrence of the genes cylA, efaA, esp, and gelE, hemolytic and gelatinase activities, capacity to form biofilm and in vitro adhesion to epithelial cells. All isolates were shown to be resistant to vancomycin and teicoplanin, as well as to two or more other antimicrobials. All isolates harbored at least one putative virulence marker, and the prevalence was as follows: esp, 87.5%; efaA, 82.5%; gelE, 70%; and cylA, 65%. The presence of 4 genes was observed in 32.5% isolates. The presence of the efaA was associated with the presence of esp, regardless of the source of the isolates. A positive association with the presence of cylA and hemolytic activity in the sheep blood agar assay was observed. No association was found for gelE and gelatinase production in the agar plate assay, for efaA and LLC-MK2 cell adhesion, and for esp and biofilm formation on polystyrene surface. These results show the presence of putative virulence genes in multiple antimicrobial resistant E. faecium isolates from different sources in a hospital setting.

Mortality and risks related to healthcare-associated infection

Healthcare-associated infections are a major cause of morbidity-mortality among hospitalized patients. The aim of this epidemiological study was to determine mortality and risks related to death in adult patients with healthcare-associated infections admitted to a teaching hospital in one year. Patient data were collected from infection medical reports. The mortality rate associated with infections was 38.4%, and it was classified as a contributing factor to deaths in 87.1% of death cases. The correlation between healthcare-associated infection and death was statistically significant among clinical patients (41.3%) presenting comorbidities related to the diagnosis (55.8%), cardiovascular infection (62.2%), pneumonia (48.9%), developing sepsis (69.0%), as well as patients who had been colonized (45.2%) and infected (44.7%) by multidrug resistance microorganisms.

Oral Candida colonization in HIV-infected patients in Londrina-PR, Brazil: antifungal susceptibility and virulence factors

INTRODUCTION Host colonization by Candida species is an important predisposing factor to candidiasis, which seems to be more frequent in human immunodeficiency virus (HIV)-infected patients. Knowledge about the distribution, antifungal susceptibility, and virulence of oral Candida isolates is important for effective management of candidiasis. METHODOLOGY Oral rinses were collected from 242 HIV-infected patients without clinical evidence of candidiasis seen at the AIDS referral center in Londrina, Brazil. Species were identified by standard phenotypic and molecular methods, and characterized in vitro according to antifungal susceptibility, cell surface hydrophobicity, biofilm formation, and enzyme activities. RESULTS Oral Candida colonization was detected in 50.4% of patients and combined use of antiretroviral therapy and protease inhibitor had a protective effect against colonization. Candida albicans (75.2%) was the most prevalent species. A high proportion of Candida spp. (39.9%) showed decreased susceptibility to fluconazole. Five isolates were resistant to nystatin. Protease and phospholipase activities were detected in 100% and 36.8% of isolates, respectively. Most isolates displayed a hydrophobic property that was associated with biofilm formation ability. CONCLUSIONS A significant number of oral Candida species exhibiting decreased susceptibility to fluconazole were isolated from colonized HIV-infected individuals. Furthermore, all isolates expressed potential virulence attributes in vitro. Given the high incidence and severity of fungal infections in HIV-infected individuals, the results of this study reinforce the importance of antifungal susceptibility testing, which contributes to therapeutic strategies and highlights the need for continuous surveillance of Candida colonization in this population.

Effect of a Metalloantibiotic Produced by Pseudomonas aeruginosa on Klebsiella pneumoniae Carbapenemase (KPC)-producing K. pneumoniae.

Multidrug-resistant organisms (MDRO) are a great problem in hospitals, where thousands of people are infected daily, with the occurrence of high mortality rates, especially in infections caused by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-producing Kpn). The challenge is to find new compounds that can control KPC producing-Kpn infections. The aim of this study was to evaluate the antibiotic activity of the F3d fraction produced by the Pseudomonas aeruginosa LV strain against clinical isolates of KPC-producing Kpn. The results showed that the minimum inhibitory concentration of F3d (62.5 µg mL(-1)), containing an organic metallic compound, killed planktonic cells of KPC-producing Kpn strains after 30 min of incubation. At the same concentration, this fraction also showed an inhibitory effect against biofilm of these bacteria after 24 h of incubation. Treatment with the F3d fraction caused pronounced morphological alterations in both planktonic and biofilm cells of the bacteria. The inhibitory effect of the F3d fraction seems to be more selective for the bacteria than the host cells, indicating its potential in the development of new drugs for the treatment of infections caused by KPC-producing Kpn and other MDRO.

Vancomycin-dependent Enterococcus faecium vanA: characterization of the first case isolated in a university hospital in Brazil

In this study, we report the characterization of a strain of Enterococcus faecium vanA, which grows only in the presence of vancomycin (VDEfm-UEL). The bacterium was isolated from the feces of a female patient who had undergone surgical treatment of Reinkes edema and was receiving intravenous vancomycin therapy for infection with methicillin/oxacillin-resistant Staphylococcus aureus, a postoperative complication. Antimicrobial dependence was further confirmed by the vancomycin E-test. VDEfm-UEL was also shown to be resistant to ampicillin, ciprofloxacin, chloramphenicol, erythromycin, levofloxacin, penicillin, rifampicin, and teicoplanin. The putative virulence genes efaA, gelE and esp were detected by PCR. The ddl gene from VDEfm-UEL was cloned and sequenced. Vancomycin dependence seems to be associated with the insertion of a nucleotide in that sequence, which results in a frame-shift mutation, introducing a premature stop codon. This is the first report of vancomycin-dependent E. faecium isolation in a university hospital in Brazil.

Antibacterial Combination of Oleoresin from Copaifera multijuga Hayne and Biogenic Silver Nanoparticles Towards Streptococcus agalactiae

BACKGROUND Streptococcus agalactiae (group B Streptococcus - GBS) remains a leading cause of neonatal infections and an important cause of invasive infections in adults with underlying conditions. METHODS This study evaluated for the first time the effect of an oleoresin collected from Copaifera multijuga Hayne (copaiba oil) alone or in combination with silver nanoparticles produced by green synthesis using Fusarium oxysporum (AgNPbio) against planktonic and sessile cells of GBS isolated from colonized women. RESULTS Copaiba oil showed a dose-dependent bactericidal activity against planktonic GBS strains, including those resistant to erythromycin and/or clindamycin. Scanning and transmission electron microscopy of GBS treated with copaiba oil revealed morphological and ultrastructural alterations, displaying disruption of the cell wall and decreased electron density due to leakage of cytoplasmic content. Copaiba oil also exhibited antibacterial activity against biofilms of GBS strains, inhibiting their formation as well as the viability of mature biofilms. In addition, the combination of copaiba oil with AgNPbio resulted in a synergistic effect against planktonic cells and biofilm formation, reducing the minimal inhibitory concentration values of both compounds. No hemolytic activity was detected for both compounds. CONCLUSION These results indicate the potential of copaiba oil, alone or in combination with AgNPbio, for the development of new alternative strategies for controlling GBS infections.

Sepsis in a university hospital: a prospective study for the cost analysis of patients' hospitalization

OBJECTIVE To estimate the cost of hospitalization of patients with severe sepsis or septic shock admitted or diagnosed in the Urgent and Emergency sector at a university hospital and followed until the clinical outcome. METHOD An epidemiological, prospective, observational study conducted in a public hospital in southern Brazil for the period of one year (August 2013 to August 2014). Sepsis notification forms, medical records and data of the cost sector were used for the collection of clinical and epidemiological data. RESULTS The sample comprised 95 patients, resulting in a total high cost of hospitalization (R

MORTALIDADE E RISCOS ASSOCIADOS A INFECÇÃO RELACIONADA À ASSISTÊNCIA À SAÚDE

3,692,421.00), and an average of R

Prevalence and antimicrobial susceptibility profile of Streptococcus agalactiae in pregnant women seen at the University Hospital of Londrina, Paraná, Brazil

38,867.60 per patient. Over half of the total value of the treatment of sepsis (R

Sepse em um hospital universitário: estudo prospectivo para análise de custo da hospitalização de pacientes

2,215,773.50) was assigned to patients who progressed to death (59.0%). The higher costs were related to discharge, diagnosis of severe sepsis, the pulmonary focus of infection and the age group of up to 59 years. CONCLUSION The high cost of the treatment of sepsis justifies investments in training actions and institution of protocols that can direct preventive actions, and optimize diagnosis and treatment in infected and septic patients. OBJETIVO Estimar o custo da internação de pacientes com sepse grave ou choque séptico admitidos ou diagnosticados no setor de Urgências e Emergências de um hospital universitário e seguidos até o desfecho clínico. MÉTODO Estudo epidemiológico, prospectivo e observacional, realizado em um hospital público do sul do Brasil, no período de 1 ano (agosto de 2013 a agosto de 2014). A coleta dos dados clínico-epidemiológicos utilizou fichas de notificação de sepse, prontuários e dados do setor de custos. Foi realizada análise de tendência central, dispersão e quartis dos custos das internações. RESULTADOS Amostra composta por 95 pacientes que totalizaram elevado custo da internação (R