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Introduction: A fast and accurate determination of methicillin resistance in Staphylococcus aureus strains is vital. This study aimed to compare the sensitivity and specificity of the cefoxitin disc diffusion (CDD) test and BD Phoenix automated system considering mecA/mecC positivity as the gold standard and to investigate the presence of Panton-Valentine leukocidin (PVL) toxin gene, a crucial virulence factor of S. aureus strains. Materials and Methods: Overall, 179 Staphylococcus aureus strains from various clinical samples were included. Antibiotic sensitivity was tested using the Phoenix automated system and by applying the Kirby-Bauer disc diffusion method for cefoxitin (30 μg). The mecA, mecC, and PVL presence was determined using the conventional multiplex polymerase chain reaction method. mecA/mecC positivity was considered as the gold standard. Statistical analysis was performed using SPSS 15.0 for Windows (Chicago, IL, USA). Results: Overall, 91 strains (50.8%) were mecA positive and identified as methicillin resistant Staphylococcus aureus (MRSA). No isolates containing the mecC gene were detected. The Phoenix automated system falsely identified six methicillin-sensitive S. aureus (MSSA) isolates, which were mecA and mecC negative as MRSA. The sensitivity and specificity of the CDD test were found to be 100% in determining MRSA, and the sensitivity and specificity the Phoenix automated system were 100% and 93.2%, respectively. The PVL positivity rate in MRSA and MSSA strains was 6.5% and 7.4%, respectively. All PVL-positive strains were isolated from the skin and soft tissues. Conclusion: The CDD test is a reliable method for routine procedures. Methicillin-sensitive strains can be determined as MRSA via the Phoenix automated system. Nevertheless, mecC-controlled MRSA should not be excluded from methods used for determining methicillin resistance. Panton-Valentine leukocidin toxin gene should be determined to enable clinicians to understand the infection severity.

Introduction: Antibiotics are lifesaving compounds that have been successful for decades. However, many pathogenic bacteria are becoming resistant to them. Cross-resistance of the macrolide-lincosamide-streptogramin B (MLSB) antibiotic classes is a major cause of increased morbidity. This study appraises the phenotypic and genotypic distribution of inducible clindamycin resistance among methicillin-resistant Staphylococcus aureus (MRSA) isolates. Materials and Methods: Erythromycin-induced resistance to clindamycin antibiotics among MRSA isolates was verified phenotypically using the Double-disk diffusion test (D-test) and genotypically by the polymerase chain reaction. Results: All MRSA isolates were resistant to erythromycin. The prevalence of iMLSB (iMLSB: inducible macrolide-lincosamide-streptogramin B) phenotype was 23.7% (9/38), macrolide streptogramin (MS) phenotype 47.4% (18/38), and cMLSB (cMLS: constitutive macrolide-lincosamide-streptogramin) phenotype 28.9% (11/38) of the isolates. The nine isolates with the iMLSB phenotype were tested for the presence of the erythromycin ribosomal methylase (erm) gene. The ermA gene was detected in five (55.6%) isolates, the ermB gene in two (22.2%) isolates, and the ermC gene in two (22.2%) isolates. Conclusion: The erm-positive isolates expressed the iMLSB phenotype, and the ermA gene was predominant. We showed that the cMLSB phenotype was prevalent among the MRSA isolates, signifying the possibility of achieving a good therapeutic outcome when clindamycin is used. The observed distribution of the erm gene explored here gives credence to the adequacy of the D-test in monitoring and testing for potential clindamycin treatment failures.

Currently multiresistant Staphylococcus aureus is one common cause of infections with high rates of morbidity and mortality worldwide, which directs scientific endeavors in search for novel antimicrobials. In this study, nine extracts from Bidens pilosa (root, stem, flower and leaves) and Annona crassiflora (rind fruit, stem, leaves, seed and pulp) were obtained with ethanol: water (7:3, v/v) and their in vitro antibacterial activity evaluated through both the agar diffusion and broth microdilution methods against 60 Oxacillin Resistant S. aureus (ORSA) strains and against S. aureus ATCC6538. The extracts from B. pilosa and A. crassiflora inhibited the growth of the ORSA isolates in both methods. Leaves of B. pilosa presented mean of the inhibition zone diameters significantly higher than chlorexidine 0.12% against ORSA, and the extracts were more active against S. aureus ATCC (p < 0.05). Parallel, toxicity testing by using MTT method and phytochemical screening were assessed, and three extracts (B. pilosa, root and leaf, and A. crassiflora, seed) did not evidence toxicity. On the other hand, the cytotoxic concentrations (CC50 and CC90) for other extracts ranged from 2.06 to 10.77 mg/mL. The presence of variable alkaloids, flavonoids, tannins and saponins was observed, even though there was a total absence of anthraquinones. Thus, the extracts from the leaves of B. pilosa revealed good anti-ORSA activity and did not exhibit toxicity. Atualmente Staphylococcus aureus multirresistente é causa comum de infecções com altas taxas de morbidade e mortalidade mundialmente, o que direciona esforços científicos na busca de novos antimicrobianos. Neste estudo, nove extratos de Bidens pilosa (raiz, caule, flor e folhas) e de Annona crassiflora (casca do fruto, caule, folha, semente e polpa) foram obtidos com etanol:água (7:3, v/v) e suas atividades antibacteriana in vitro avaliadas através de difusão em agar e microdiluição em caldo contra 60 cepas de Oxacillin Resistant S. aureus (ORSA) e contra S. aureus ATCC 6538. Os extratos de B. pilosa e A. crassiflora inibiram o crescimento dos isolados ORSA em ambos os métodos. O extrato da folha de B. pilosa apresentou média dos diâmetros dos halos de inibição significativamente maior que a clorexidina 0,12%, contra os isolados ORSA, e os extratos foram mais ativos contra S. aureus ATCC (p < 0,05). Paralelamente, teste de toxicidade pelo método MTT e triagem fitoquímica foram avaliadas, e três extratos (raiz e folha de B. pilosa e semente de A. crassiflora) não apresentaram toxicidade. Por outro lado, as concentrações citotóxicas (CC50 e CC90) para os outros extratos variaram de 2,06 a 10,77 mg/mL. Observou-se variável presença de alcalóides, flavonóides, taninos e saponinas, apesar de total ausência de antraquinonas. Portanto, os extratos das folhas de B. pilosa revelaram boa atividade anti-ORSA e não exibiram toxicidade.

Introduction Methicillin resistant Staphylococcus aureus has emerged as an important pathogen in nosocomial and community acquired infections. Accurate and rapid identification of MRSA in clinical specimens is essential for timely decision of effective antimicrobial chemotherapy. Aim The present study was conducted to compare efficacy of four conventional phenotypic methods, with mec- A based polymerase chain reaction (PCR) for MRSA identification. Materials and methods Methicillin resistance was determined in 200 S.aureus isolates by oxacillin disc diffusion, cefoxitin disc diffusion, Oxacillin Resistance Screening Agar and E-test. The results were compared with mec-A based PCR. Results Among 200 S.aureus isolates 62 (31%) were positive for mec-A gene by PCR. Cefoxitin disc diffusion, Oxacillin Resistance Screening Agar and E-test showed 100% specificity. Oxacillin disc diffusion had lowest sensitivity (82.5%) and specificity (98.5%) among all. The conventional methods take more time than PCR for diagnosing MRSA. Linezolid, Vancomycin & Dalfopristin were the highly sensitive drugs against MRSA isolates. Conclusion Cefoxitin disc diffusion, is rapid, simple and cheaper, hence can be used routinely as an alternative to PCR for detection of MRSA in resource constraint laboratories.

Arbekacin is an aminoglycoside used in Japan for treating infections caused by gentamicin and oxacillin-resistant S. aureus (ORSA). The objective of this study was to determine the in vitro antimicrobial activity of arbekacin against 454 clinical isolates of ORSA. The isolates were consecutively collected between January and July, 2000, from patients hospitalized in 8 Brazilian medical centers. The antimicrobial susceptibility testing was performed by disk diffusion method according to NCCLS recommendations. The vast majority of the isolates, 453 strains (99.8%), were considered susceptible to arbekacin based on the criteria proposed by the Requirements for Antibiotic Products of Japan. Only 1 isolate (0.2%) was classified as resistant. On the other hand, high rates of resistance were demonstrated for other aminoglycosides, such as gentamicin (97.6% resistance) and amikacin (97.0% resistance). Resistance rate was also high for ciprofloxacin (98.0%). All isolates were considered susceptible to vancomycin. The excellent in vitro antimicrobial activity of arbekacin demonstrated in this study indicates that this antimicrobial agent may play an important role in the treatment of severe ORSA infections, especially those that show poor clinical response with vancomycin monotherapy. Since the aminoglycosides should not be used as monotherapy to treat Gram positive infections, further studies evaluating in vitro and in vivo synergistic activity of arbekacin combinations are necessary to clarify the clinical role of this aminoglycoside. Universidade Federal de São Paulo (UNIFESP) Division of Infectious Diseases Special Clinical Microbiology Laboratory UNIFESP, Division of Infectious Diseases Special Clinical Microbiology Laboratory SciELO