Clonal dissemination of vancomycin-resistant Enterococcus faecium ST412 in a Brazilian region

ABSTRACT Vancomycin-resistant Enterococcus faecium (VREfm) has emerged as an important global nosocomial pathogen, and this trend is associated with the spread of high-risk clones. Here, we determined the genetic and phenotypic features of 93 VREfm isolates that were obtained from patients in 13 hospitals in Vitória, Espírito Santo, Brazil, during 2012-2013. All the isolates were vancomycin-resistant and harbored the vanA gene. Only 6 (6.5%) of the VREfm isolates showed the ability to form biofilm. The 93 isolates analyzed belong to a single pulsed-field gel electrophoresis lineage and presented six subtypes. MLST genotyping showed that all VREfm belonged to ST412 (the high-risk clone, hospital-adapted). The present study describes the dissemination of ST412 clone in the local hospitals. The clonal spread of these ST412 isolates in the area we analyzed as well as other hospitals in southeastern Brazil supports the importance of identifying and controlling the presence of these microorganisms in health care-related services.

Evaluation of the synergistic potential of vancomycin combined with other antimicrobial agents against methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus spp strains

Methicillin-resistant Staphylococcus aureus (MRSA) and coagulase-negative Staphylococcus spp (CNS) are the most common pathogens that cause serious long term infections in patients. Despite the existence of new antimicrobial agents, such as linezolid, vancomycin (VAN) remains the standard therapy for the treatment of infections caused by these multidrug-resistant strains. However, the use of VAN has been associated with a high frequency of therapeutic failures in some clinical scenarios, mainly with decreasing concentration of VAN. This work aims to evaluate the synergic potential of VAN plus sulfamethoxazole/trimethoprim (SXT), VAN plus rifampin (RIF) and VAN plus imipenem (IPM) in sub-minimum inhibitory concentrations against 22 clinical strains of MRSA and CNS. The checkerboard method showed synergism of VAN/RIF and VAN/SXT against two and three of the 22 strains, respectively. The combination of VAN with IPM showed synergistic effects against 21 out of 22 strains by the E-test method. Four strains were analyzed by the time-kill curve method and synergistic activity was observed with VAN/SXT, VAN/RIF and especially VAN/IPM in sub-inhibitory concentrations. It would be interesting to determine if synergy occurs in vivo. Evidence of in vivo synergy could lead to a reduction of the standard VAN dosage or treatment time.

Avaliação de métodos comumente usados em laboratórios para a determinação da suscetibilidade à oxacilina entre amostras de Staphylococcus sp, isoladas de um hospital de Vitória, Estado do Espírito Santo / Evaluation of methods commonly used in laboratories to determine the susceptibility to oxacillin among Staphylococcus sp samples isolated from a hospital in Vitória, State of Espírito Santo

INTRODUÇÃO: O gênero Staphylococcus é de grande importância devido a sua alta prevalência em infecções hospitalares e por apresentar taxas elevadas de resistência a oxacilina e a outros antimicrobianos. Assim, a avaliação da acurácia dos métodos fenotípicos usados para determinação do perfil de suscetibilidade a antimicrobianos é essencial para garantir a escolha da terapia mais adequada. MÉTODOS: Foram usadas 114 amostras de Staphylococcus sp (53 S. aureus e 61 SCN) na avaliação da acurácia dos métodos de difusão de disco, microdiluição em agar, ágar triagem oxacilina e sistema automatizado em comparação com a PCR para verificação da resistência a oxacilina. RESULTADOS: O gene mecA foi detectado em 48 (42,1 por cento) amostras e 27 (23,7 por cento) amostras apresentaram discrepância de resultados em pelo menos um dos métodos (74,1 por cento SCN, 25,9 por cento S. aureus). Para S. aureus, com exceção do Microscan Walkaway, todos os métodos apresentaram 100 por cento de especificidade e sensibilidade. Já para os SCN, o sistema automatizado e o disco de cefoxitina apresentaram menor acurácia. CONCLUSÕES: O uso de dois métodos deve ser a melhor opção para a melhora da acurácia, principalmente quando o laboratório de diagnóstico utiliza somente sistema automatizado ou teste de difusão do disco de oxacilina. A associação destes métodos com outros apresentaram praticamente 100 por cento de sensibilidade e especificidade em nosso estudo.

INTRODUCTION: The genus Staphylococcus is of great importance because of its high prevalence in hospital infections and because it presents high rates of resistance to oxacillin and other antimicrobials. Thus, evaluation of the accuracy of the phenotypic methods that are used to determine the profile of antimicrobial resistance is essential to ensure that the most appropriate therapy is chosen. METHODS: One hundred and fourteen strains of Staphylococcus sp (53 S. aureus and 61 CNS) were used to evaluate the accuracy of the methods of disk diffusion, agar microdilution, oxacillin screening agar and automated systems, in comparison with PCR for investigating resistance to oxacillin. RESULTS: The mecA gene was detected in 48 strains (42.1 percent), and 27 strains (23.7 percent) showed discrepant results in at least one of the methods (74.1 percent of CNS, 25.9 percent of S. aureus). For S. aureus, with the exception of the Microscan Walkaway, all the methods showed 100 percent specificity and sensitivity. In relation to CNS, the automated system and cefoxitin disk had lower accuracy. CONCLUSIONS: Use of two methods should be the best option for improved accuracy, especially when the diagnostic laboratory only uses an automated system or oxacillin disk diffusion test. Combination of these methods with others presented almost 100 percent sensitivity and specificity in our study.

Heterogeneous resistance to vancomycin and teicoplanin among Staphylococcus spp. isolated from bacteremia

This study evaluated the BHIA screening method with 4 or 6 mug/mL of vancomycin to detect glycopeptides heteroresistant staphylococci strains isolated from bacteremia. A total of 213 staphylococci strains were isolated from 106 patients between October/2001 and November/2002 in a tertiary hospital in Rio de Janeiro city. Fifty-seven (53.8 percent) patients presented Staphylococcus aureus, while coagulase-negative staphylococci (CNS) were isolated from 49 (46.2 percent). Resistance rates for oxacillin of 26.3 percent and 81.6 percent were found for the staphylococci isolates, respectively. Thirteen CNS isolated from nine (8.5 percent) patients grew on agar screening with 4 mug/mL of vancomycin and showed heterogeneous profiles of resistance for vancomycin and teicoplanin by the population analysis profile method. Only 30.8 percent of them grew at the concentration 6 mug/mL. Bacterial infection and use of antimicrobial therapy were common among these patients. Alert about the emergence of oxacillin-resistant staphylococci presenting heteroresistance to glycopeptides is important in order to achieve judicious use of antimicrobials. Vancomycin agar screening test could help to confirm the presence of these isolates in hospitals.