Marcia Regina Eches Perugini

Commensal Streptococcus agalactiae isolated from patients seen at University Hospital of Londrina, Paraná, Brazil: capsular types, genotyping, antimicrobial susceptibility and virulence determinants

BackgroundStreptococcus agalactiae or Group B Streptococci (GBS) have the ability to access various host sites, which reflects its adaptability to different environments during the course of infection. This adaptation is due to the expression of virulence factors that are involved with survival, invasion and bacterial persistence in the host. This study aimed to characterize GBS isolates from women of reproductive age seen at University Hospital of Londrina, according to capsular typing, genetic relatedness, antimicrobial susceptibility profile and occurrence of virulence determinants.ResultsA total of 83 GBS isolates were enrolled in this study. Capsular types Ia (42.2%), II (10.8%), III (14.5%) and V (30.1%) were identified in most GBS. One isolate each was classified as type IX and non-typeable.A total of 15 multiple locus variable number of tandem repeat analysis (MLVA) types were identified among the isolates, seven were singletons and eight were represented by more than four isolates. All isolates were susceptible to penicillin, ampicillin, cefepime, cefotaxime, chloramphenicol, levofloxacin and vancomycin. Resistance to erythromycin and clindamycin was observed in 19.3 and 13.3% of isolates, respectively. All isolates resistant to clindamycin were simultaneously resistant to erythromycin and were distributed in the capsular types III and V. One isolate showed the constitutive macrolide-lincosamide-streptogramin B (cMLSB) phenotype and ten showed the inducible MLSB (iMLSB) phenotype. The mechanism of resistance to erythromycin and clindamycin more prevalent among these isolates was mediated by the gene ermA, alone or in combination with the gene ermB. The isolates displaying resistance only to erythromycin belonged to capsular type Ia, and showed the M phenotype, which was mediated by the mefA/E gene. All isolates harbored the gene hylB and at least one pilus variant, PI-1, PI-2a or PI-2b. Although cylE was observed in all GBS, four isolates were classified as gamma-hemolytic and carotenoid pigment non-producers.ConclusionsOur results indicate the potential virulence of commensal GBS isolates, reinforcing the need for continued screening for this bacterium to prevent infections. The distribution of capsular and pili antigens, and MLVA profiles was also identified, which may contribute to the development of new strategies for the prevention and treatment of GBS infection.

Effect of Eugenol against Streptococcus agalactiae and Synergistic Interaction with Biologically Produced Silver Nanoparticles

Streptococcus agalactiae (group B streptococci (GBS)) is an important infections agent in newborns associated with maternal vaginal colonization. Intrapartum antibiotic prophylaxis in GBS-colonized pregnant women has led to a significant reduction in the incidence of early neonatal infection in various geographic regions. However, this strategy may lead to resistance selecting among GBS, indicating the need for new alternatives to prevent bacterial transmission and even to treat GBS infections. This study reported for the first time the effect of eugenol on GBS isolated from colonized women, alone and in combination with silver nanoparticles produced by Fusarium oxysporum (AgNPbio). Eugenol showed a bactericidal effect against planktonic cells of all GBS strains, and this effect appeared to be time-dependent as judged by the time-kill curves and viability analysis. Combination of eugenol with AgNPbio resulted in a strong synergistic activity, significantly reducing the minimum inhibitory concentration values of both compounds. Scanning and transmission electron microscopy revealed fragmented cells and changes in bacterial morphology after incubation with eugenol. In addition, eugenol inhibited the viability of sessile cells during biofilm formation and in mature biofilms. These results indicate the potential of eugenol as an alternative for controlling GBS infections.

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.

Detection of blaVIM-7 in an extensively drug-resistant Pseudomonas aeruginosa isolate belonging to ST1284 in Brazil

We described for the first time an extensively drug-resistant Pseudomonas aeruginosa isolate belonging to ST1284 carrying a plasmid-mediated blaVIM-7 in Brazil. The blaVIM-7 was harbored by an integron that also carried aacA4 and blaOXA-46. Multiple virulence factors were also detected.

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.

Antimicrobial activity evaluation and comparison of methods of susceptibility for Klebsiella pneumoniae carbapenemase (KPC)-producing Enterobacter spp. isolates

The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest® (bioMérieux), Vitek 2® automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2® automated system was the method most similar compared to the broth microdilution. Therefore, is important to evaluate the performance of new methods in comparison to the reference method, broth microdilution.

Comparison of HRM analysis and three REP-PCR genomic fingerprint methods for rapid typing of MRSA at a Brazilian hospital

INTRODUCTION Infections caused by multidrug-resistant bacteria are increasingly common and represent a serious problem for public health. Staphylococcus aureus is one of the major agents of infections, and methicillin-resistant S. aureus (MRSA) has spread worldwide. The aim of this study was to phenotypically and genotypically characterize 55 MRSAs isolated in the University Hospital of Londrina, Paraná, Brazil, during 2010. METHODOLOGY Bacterial isolates were characterized based on their antimicrobial susceptibility profile, biofilm production capacity, and staphylococcal chromosome cassette mec (SCCmec) type. Determination of clonal groups was performed by polymerase chain reaction using the RW3A, JB1, and BOX A1R primers and high-resolution melting (HRM) analysis. RESULTS The majority of isolates harbored SCCmec type II. SCCmec III, characteristic of the Brazilian endemic clone, was observed in four strains. Only two isolates harbored SCCmec type IV, which is common in community-acquired MRSA strains. Most isolates also showed resistance to more than four of the tested antimicrobials, and 30 isolates exhibited the ability to produce biofilm. DNA polymorphism analysis showed a higher discriminatory power for the JB1 primer, but RW3A revealed several clonal groups of MRSA with similar genotypic and phenotypic characteristics. HRM analysis showed eight different sequence types. CONCLUSIONS These results are important for epidemiological studies involving MRSA infections.

MORTALIDADE E RISCOS ASSOCIADOS A INFECÇÃO RELACIONADA À ASSISTÊNCIA À SAÚDE

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.

Association of plasmid typing to biotyping and antibiotyping in the characterization of outbreaks by Acinetobacter baumannii

Durante um surto ocorrido de abril a setembro de 1994 em um Hospital Universitario, dezesseis cepas de Acinetobacter baumannii foram isoladas de pacientes e uma de solucao enteral. Nos posteriormente analizamos as cepas isoladas durante o surto pelos seguintes metodos de tipagem : perfil de DNA plasmidial, perfil de antibiograma e biotipagem. Dois padroes de tipagem foram identificados pela analise do perfil plasmidial. Doze cepas foram caracterizadas como sendo do biotipo 2, e cinco do biotipo 19. O padrao de sensibilidade a amicacina e a carbenicilina possibilitou a classificacao das cepas em dois grupos. Os resultados demonstraram que estes tres metodos de tipagem associados possibilitaram a diferenciacao do que primeiramente foi considerado como um unico surto, em dois surtos aparentemente nao relacionados.