Origanum vulgare essential oil and carvacrol: natural alternatives against resistant bacteria / Óleo essencial de origanum vulgare e carvacrol: alternativas naturais contra bactérias resistentes / El aceite esencial de origanum vulgare y el carvacrol: alternativos naturales contra las bacterias resistentes

Bacteria that are resistant to several antibiotics are a serious One Health problem, as new alternatives for treatment do not appear at the same speed. Thus, the aim of this work was to carry out a survey of studies involving the activity of the essential oil of O. vulgare and its isolated compound carvacrol on antibiotic-resistant bacteria. To this end, a qualitative review of the literature was carried out in the PubMed database from 2015 to 2020. Both for the essential oil and for the isolated compound, the inhibitory action extends to strains often associated with difficult-to-treat infections such as oxacillin and vancomycin-resistant Staphylococcus aureus, ß-lactamase-producing strains, carbapenemases, among others. The point that distinguishes the studies is the type of methodology used in the tests, with studies with carvacrol more directed towards mechanisms of molecular action and application in cells and animals, while those with oils are more preliminary. Although these substances have potential to control resistant bacteria, more research is needed to enable their use.

Bactérias resistentes a vários antibióticos são um grave problema para a Saúde Única, pois novas alternativas de tratamento não aparecem na mesma velocidade. Assim, o objetivo deste trabalho foi realizar um levantamento de estudos envolvendo a atividade do óleo essencial de O. vulgare e seu composto isolado, carvacrol, sobre bactérias resistentes a antibióticos. Para tanto, foi realizada uma revisão qualitativa da literatura na base de dados PubMed no período de 2015 a 2020. Tanto para o óleo essencial quanto para o composto isolado, a ação inibitória se estende a cepas frequentemente associadas a infecções de difícil tratamento como Staphylococcus aureus resistente à oxacilina e vancomicina, cepas produtoras de ß-lactamase, carbapenemases, entre outras. O ponto que diferencia os estudos é o tipo de metodologia utilizada nos testes, sendo os estudos com carvacrol mais direcionados para mecanismos de ação molecular e aplicação em células e animais, enquanto os com óleos são mais preliminares. Embora essas substâncias tenham potencial para controlar bactérias resistentes, mais pesquisas são necessárias para viabilizar seu uso.

Las bacterias resistentes a diversos antibióticos son un grave problema para la Sanidad Única, ya que las nuevas alternativas de tratamiento no aparecen a la misma velocidad. Así pues, el objetivo de este trabajo fue realizar una encuesta sobre los estudios relativos a la actividad del aceite esencial de O. vulgare y su compuesto aislado, el carvacrol, sobre las bacterias resistentes a los antibióticos. Para ello, se realizó una revisión bibliográfica cualitativa en la base de datos PubMed en el periodo comprendido entre 2015 y 2020. Tanto para el aceite esencial como para el compuesto aislado, la acción inhibidora se extiende a cepas frecuentemente asociadas a infecciones de difícil tratamiento como el Staphylococcus aureus resistente a la oxacilina y a la vancomicina, cepas productoras de ß-lactamasas, carbapenemasas, entre otras. El punto que diferencia los estudios es el tipo de metodología utilizada en las pruebas, siendo los estudios con carvacrol más dirigidos a mecanismos de acción molecular y aplicación en células y animales, mientras que los de aceites son más preliminares. Aunque estas sustancias tienen potencial para controlar las bacterias resistentes, es necesario seguir investigando para que su uso sea viable.

Antimicrobial activity of dalbavancin and comparators against Staphylococcus aureus causing pneumonia in patients with and without cystic fibrosis.

The activities of dalbavancin and comparator agents were evaluated against Staphylococcus aureus isolated from the lower respiratory tract of cystic fibrosis (CF) and non-CF patients with pneumonia. Bacterial isolates (n = 357) were collected from CF patients in 36 medical centers worldwide (2018-2019) and susceptibility tested using reference broth microdilution. Susceptibility results from these isolates were compared with those for 725 S. aureus isolates consecutively collected from non-CF patients with pneumonia from the same medical centers over the same period. Only isolates determined to be the probable cause of pneumonia were included in the study. Susceptibility profiles were very similar among isolates from CF and non-CF patients. Dalbavancin exhibited potent activity (MIC50/90, 0.03/0.03 mg/L) and complete coverage (100.0% susceptibility) against isolates from CF and non-CF patients. Ceftaroline (MIC50/90, 0.25/1 mg/L) was active against 97.8% and 98.1% of isolates from CF and non-CF patients, respectively. Oxacillin resistance (MRSA) rates were 27.7% among CF and 28.7% among non-CF patients. Among MRSA isolates from CF/non-CF patients (n = 99/208), susceptibility to ceftaroline, clindamycin, levofloxacin, and tetracycline were 91.9%/93.3%, 58.6%/64.4%, 40.4%/29.3%, and 83.8%/89.4%, respectively. Dalbavancin demonstrated high potency against S. aureus from CF and non-CF patients and may represent a valuable treatment option for CF patients with MRSA pulmonary infection.

Neonatal septicemia at intensive care unit, Ayder Comprehensive Specialized Hospital, Tigray, North Ethiopia: Bacteriological profile, drug susceptibility pattern, and associated factors.

BACKGROUND: Neonatal septicemia is a life threatening medical emergency that requires timely detection of pathogens with urgent rational antibiotics therapy. METHODS: A cross-sectional study was conducted between March 2017 to September 2018 among 317 septicemia suspected neonates at neonatal intensive care unit, Ayder Comprehensive Specialized Hospital, Mekelle, Tigray, North Ethiopia. A 3 mL of blood was collected from each participant. Identification of bacterial species was done using the standard microbiological techniques. Antibiotic sensitivity test was done using disk diffusion method. Data were entered and analyzed using computer software SPSS version 22. Bivariate and multivariate regression analysis was applied to determine the association between variables. RESULTS: Of the 317 (190 male and 127 female) neonates, 116 (36.6%) were found to be with culture proven septicemia. Klebsiella species were the predominant etiologic agents. Length of hospital stay (AOR (adjusted odds ratio) = 3.65 (2.17-6.13), p < 0.001) and low birth weight (AOR = 1.64 (1.13-2.78), p = 0.04) were the factors associated with neonatalsepticemia. Most isolates showeda frightening drug resistance rate to the commonly used antimicrobial drugs. K. pneumoniae, E. coli, Enterobacter and Citrobacter species were 57% to100% resistant to ceftazidime, ceftriaxone, gentamycin, amoxacillin-clavulunic acid and ampicillin. All, 9 (100%) isolates of S. aureus were resistant to oxacilline, ampicillin,erythromycin and gentamycin. Furthermore, 55.6% S. aureus isolates were Methicillin Resistant Staphylococcus aureus. CONCLUSION: Neonaltal septicemia is found to be significantly high in the present study. As most of the isolates are potentially related to hospital acquired infections, prevention and control policy should have to be more strengthening in the neonatal intensive care unit.

Antibiotic resistance among bacterial conjunctival pathogens collected in the Antibiotic Resistance Monitoring in Ocular Microorganisms (ARMOR) surveillance study.

The Antibiotic Resistance Monitoring in Ocular Microorganisms (ARMOR) surveillance study evaluates in vitro antibiotic resistance among Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Streptococcus pneumoniae, Pseudomonas aeruginosa, and Haemophilus influenzae isolates from ocular infections. Here we report resistance rates and trends among conjunctival-sourced ocular isolates collected across the US from 2009 through 2016. A total of 1198 conjunctival isolates (483 S. aureus, 305 CoNS, 208 H. influenzae, 118 S. pneumoniae, and 84 P. aeruginosa) were collected from patients with presumed bacterial conjunctivitis from 57 sites across 40 states. A large proportion of staphylococci demonstrated resistance to oxacillin and azithromycin, while resistance was low against the majority of antibiotics tested for S. pneumoniae, P. aeruginosa, and H. influenzae. Multidrug resistance (≥3 antibiotic classes) was found in 30.2% of S. aureus and 39.0% of CoNS isolates, and methicillin resistance more than doubled the rate of multi-drug resistance (methicillin-resistant S. aureus [MRSA], 76.5%; methicillin-resistant CoNS isolates, 72.8%). There was a pattern of increasing mean percent resistance with increasing age by decade of life among S. aureus, MRSA, and CoNS (P≤0.038). Over the eight-year study period, there were small yet significant decreases in resistance rates among S. aureus to azithromycin, ciprofloxacin, tobramycin, trimethoprim, and oxacillin (P≤0.003), and among CoNS and P. aeruginosa (both P<0.05) to ciprofloxacin. These data indicate that antibiotic resistance is high, but did not increase, among conjunctival-sourced isolates collected in the US from 2009 through 2016. For certain antibiotic/pathogen combinations, there was a trend of decreased resistance, including a decrease in oxacillin resistance among S. aureus.

Characterizing non-linear effects of hospitalisation duration on antimicrobial resistance in respiratory isolates: an analysis of a prospective nationwide surveillance system.

OBJECTIVES: Our objective was to systematically study the influence of length of hospital stay on bacterial resistance in relevant respiratory tract isolates. METHODS: Using prospective epidemiological data from the National Swiss Antibiotic Resistance Surveillance System, susceptibility testing results for respiratory isolates retrospectively retrieved from patients hospitalised between 2008 and 2014 were compiled. Generalized additive models were used to illustrate resistance rates relative to hospitalisation duration and to adjust for co-variables. RESULTS: In all, 19 622 isolates of six relevant and predominant species were included. Resistance patterns for the predominant species showed a species-specific and antibiotic-resistance-specific profile in function of hospitalisation duration. The oxacillin resistance profile in Staphylococcus aureus isolates was constantly increasing (monophasic). The pattern of resistance to cefepime in Pseudomonas aeruginosa was biphasic with a decreasing resistance rate for the first 5 days of hospitalisation and an increase for days 6-30. A different biphasic pattern occurred in Escherichia coli regarding amoxicillin-clavulanic acid resistance: odds/day increased for the first 7 days of hospitalisation and then remained stable for days 8-30. In the adjusted models epidemiological characteristics such as age, ward type, hospital type and linguistic region were identified as relevant co-variables for the resistance rates. The contribution of these confounders was specific to the individual species/antibiotic resistance models. CONCLUSIONS: Resistance rates do not follow a dichotomic pattern (early versus late nosocomial) as suggested by current hospital-acquired pneumonia treatment guidelines. Duration of hospitalisation rather appears to have a more complex and non-linear relationship with bacterial resistance in hospital-acquired pneumonia, also depending on host and environmental factors.

A Retrospective Analysis of Treatment and Clinical Outcomes among Patients with Methicillin-Susceptible Staphylococcus aureus Bloodstream Isolates Possessing Detectable mecA by a Commercial PCR Assay Compared to Patients with Methicillin-Resistant Staphylococcus aureus Bloodstream Isolates.

mecA-positive Staphylococcus aureus isolates phenotypically susceptible to cefoxitin (mecA-methicillin-sensitive S. aureus [MSSA]) have been identified. We describe the treatment and outcomes among patients with mecA-MSSA bloodstream infections (BSI) and MRSA BSI matched 1:1 for age, BSI origin, and BSI type (n = 17 per group). Compared to MRSA BSI patients, mecA-MSSA BSI patients more often experienced clinical failure (58.8% and 11.8%, P = 0.010), driven largely by persistent bacteremia (35.3% and 11.8%). mecA-MSSA BSI patients may be at higher risk for poor clinical outcomes.

Action of antibiotic oxacillin on in vitro growth of methicillin-resistant Staphylococcus aureus (MRSA) previously treated with homeopathic medicines.

BACKGROUND: Resistance to antibiotics is a major public health concern worldwide. New treatment options are needed and homeopathy is one such option. We sought to assess the effect of the homeopathic medicine Belladonna (Bell) and a nosode (biotherapy) prepared from a multi-drug resistant bacterial species, methicillin-resistant Staphylococcus aureus (MRSA), on the same bacterium. METHODS: Bell and MRSA nosode were prepared in 6cH and 30cH potencies in 30% alcohol and sterile water, according to the Brazilian Homeopathic Pharmacopeia and tested on MRSA National Collection of Type Cultures (NCTC) 10442. We assessed in vitro bacterial growth, deoxyribonuclease (DNAase) and hemolysin activity, and in vitro bacterial growth in combination with oxacillin (minimum inhibitory concentration - MIC). All values were compared to control: 30% alcohol and water. RESULTS: In vitro growth of MRSA was statistically significantly inhibited in the presence of Bell and nosode 6cH and 30cH compared to controls (p < 0.0001); and with combination of Bell or nosode 6cH and 30cH and oxacillin (p < 0.001). Bell 30cH and nosode 6cH and 30cH significantly decreased bacterial DNAse production (p < 0.001) and reduced red blood cell lysis. CONCLUSIONS: Cultures of MRSA treated with Belladonna or MRSA nosode exhibited reduced growth in vitro, reduced enzymatic activity and became more vulnerable to the action of the antibiotic oxacillin. Further studies are needed on the biomolecular basis of these effects.

Redeploying ß-Lactam Antibiotics as a Novel Antivirulence Strategy for the Treatment of Methicillin-Resistant Staphylococcus aureus Infections.

Innovative approaches to the use of existing antibiotics is an important strategy in efforts to address the escalating antimicrobial resistance crisis. We report a new approach to the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections by demonstrating that oxacillin can be used to significantly attenuate the virulence of MRSA despite the pathogen being resistant to this drug. Using mechanistic in vitro assays and in vivo models of invasive pneumonia and sepsis, we show that oxacillin-treated MRSA strains are significantly attenuated in virulence. This effect is based primarily on the oxacillin-dependent repression of the accessory gene regulator quorum-sensing system and altered cell wall architecture, which in turn lead to increased susceptibility to host killing of MRSA. Our data indicate that ß-lactam antibiotics should be included in the treatment regimen as an adjunct antivirulence therapy for patients with MRSA infections. This would represent an important change to current clinical practice for treatment of MRSA infection, with the potential to significantly improve patient outcomes in a safe, cost-effective manner.

A case of atypical nocardia asteroides sclerouveitis resistant to fourth-generation fluoroquinolones / Um caso atípico de esclero-uveíte por Nocardia Asteroides resistente a fluorquinolona de quarta-geração

ABSTRACT We describe an unusual case of Nocardia spp scleritis in a health girl resistant to topical fourth-generation fluoroquinolones. Clinically, there was only partial response of the scleritis to initial therapy. Treatment was changed to meropenem intravenously and topical amikacin. Following several weeks of antibiotic treatment, the patient's infection resolved but her vision was reduced to no light perception. Nocardia asteroides must be considered as a possible agent in cases of necrotizing scleritis in patients without a clear source. Antibiotic sensitivity testing has a definitive role in view of the resistance to these new medications.

RESUMO Nós descrevemos um raro caso de esclerite por Nocardia spp em uma criança sadia resistente a utilização tópica de fluorquinolona de quarta-geração. Clinicamente, a paciente apresentou apenas uma resposta parcial do quadro de esclerite a terapêutica inicial. O tratamento foi então modificado para meropenem intravenoso e amicacina tópica. Após várias semanas de tratamento com antibiótico, o quadro infeccioso regrediu porém a visao da pacientes evoluiu para perda da percepção luminosa. Em casos de esclerite necrotizante em pacientes sem fatores de risco aparente é necessário considerer a Nocardia Asteroides como possível agente causador. Os testes de sensibilidade medicamentosa apresentam importância significativa em virtude do aparecimento de resistência aos novos medicamentos.

Comparison of cefazolin versus oxacillin for treatment of complicated bacteremia caused by methicillin-susceptible Staphylococcus aureus.

Contrary to prior case reports that described occasional clinical failures with cefazolin for methicillin-susceptible Staphylococcus aureus (MSSA) infections, recent studies have demonstrated no difference in outcomes between cefazolin and antistaphylococcal penicillins for the treatment of MSSA bacteremia. While promising, these studies described low frequencies of high-inoculum infections, such as endocarditis. This retrospective study compares clinical outcomes of cefazolin versus oxacillin for complicated MSSA bacteremia at two tertiary care hospitals between January 2008 and June 2012. Fifty-nine patients treated with cefazolin and 34 patients treated with oxacillin were included. Osteoarticular (41%) and endovascular (20%) sources were the predominant sites of infection. The rates of clinical cure at the end of therapy were similar between cefazolin and oxacillin (95% versus 88%; P=0.25), but overall failure at 90 days was higher in the oxacillin arm (47% versus 24%; P=0.04). Failures were more likely to have received surgical interventions (63% versus 40%; P=0.05) and to have an osteoarticular source (57% versus 33%; P=0.04). Failures also had a longer duration of bacteremia (7 versus 3 days; P=0.0002), which was the only predictor of failure. Antibiotic selection was not predictive of failure. Rates of adverse drug events were higher in the oxacillin arm (30% versus 3%; P=0.0006), and oxacillin was more frequently discontinued due to adverse drug events (21% versus 3%; P=0.01). Cefazolin appears similar to oxacillin for the treatment of complicated MSSA bacteremia but with significantly improved safety. The higher rates of failure with oxacillin may have been confounded by other patient factors and warrant further investigation.

Bioluminescence and 19F magnetic resonance imaging visualize the efficacy of lysostaphin alone and in combination with oxacillin against Staphylococcus aureus in murine thigh and catheter-associated infection models.

Staphylococci are the leading cause of hospital-acquired infections worldwide. Increasingly, they resist antibiotic treatment owing to the development of multiple antibiotic resistance mechanisms in most strains. Therefore, the activity and efficacy of recombinant lysostaphin as a drug against this pathogen have been evaluated. Lysostaphin exerts high levels of activity against antibiotic-resistant strains of Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA). The therapeutic value of lysostaphin has been analyzed in two different clinically relevant in vivo models, a catheter-associated infection model and a thigh infection model. We infected mice with luciferase-expressing S. aureus Xen 29, and the efficacies of lysostaphin, vancomycin, oxacillin, and combined lysostaphin-oxacillin were investigated by determining numbers of CFU, detecting bioluminescent signals, and measuring the accumulation of perfluorocarbon emulsion at the site of infection by (19)F magnetic resonance imaging. Lysostaphin treatment significantly reduced the bacterial burden in infected thigh muscles and, after systemic spreading from the catheter, in inner organs. The efficiency of lysostaphin treatment was even more pronounced in combinatorial therapy with oxacillin. These results suggest that recombinant lysostaphin may have potential as an anti-S. aureus drug worthy of further clinical development. In addition, both imaging technologies demonstrated efficacy patterns similar to that of CFU determination, although they proved to be less sensitive. Nonetheless, they served as powerful tools to provide additional information about the course and gravity of infection in a noninvasive manner, possibly allowing a reduction in the number of animals needed for research evaluation of new antibiotics in future studies.

Interplay of antibiotics and bacterial inoculum on suture-associated biofilms.

BACKGROUND: Biofilms are often antibiotic resistant, and it is unclear if prophylactic antibiotics can effectively prevent biofilm formation. Experiments were designed to test the ability of high (bactericidal) concentrations of ampicillin (AMP), vancomycin (VAN), and oxacillin (OXA) to prevent formation of suture-associated biofilms initiated with low (10(4)) and high (10(7)) numbers of Staphylococcus aureus. MATERIALS AND METHODS: S. aureus biofilms were cultivated overnight on silk suture incubated in biofilm growth medium supplemented with bactericidal concentrations of AMP, VAN, or OXA. Standard microbiological methods were used to quantify total numbers of viable suture-associated S. aureus. Crystal violet staining followed by spectroscopy was used to quantify biofilm biomass, which includes bacterial cells plus matrix components. To observe the effects of antibiotics on the microscopic appearance of biofilm formation, biofilms were cultivated on glass slides, then stained with fluorescent dyes, and observed by confocal microscopy. RESULTS: In the presence of a relatively low inoculum (10(4)) of S. aureus cells, bactericidal concentrations of AMP, VAN, or OXA were effective in preventing development of suture-associated biofilms. However, similar concentrations of these antibiotics were typically ineffective in preventing biofilm development on sutures inoculated with 10(7)S. aureus, a concentration relevant to contaminated skin. Confocal microscopy confirmed that bactericidal concentrations of AMP, VAN, or OXA inhibited, but did not prevent, development of S. aureus biofilms. CONCLUSION: Bactericidal concentrations of AMP, VAN, or OXA inhibited formation of suture-associated biofilms initiated with low numbers (10(4)), but not high numbers (10(7)), of S. aureus cells.

Evaluación de la susceptibilidad a glicopéptidos en cepas de Staphylococcus aureus resistentes a oxacilina / Evaluation of susceptibility to glycopeptides in oxacillin resistant Staphylococcus aureus strains

Candida albicans y Candida dubliniensis presentan una estrecha relación filogenética. La similitud de estas especies puede hacer que en un laboratorio microbiológico se identifique en forma errónea C. dubliniensis como C. albicans. El objetivo de esta investigación fue evaluar diversos métodos fenotípicos para la diferenciación entre Candida dubliniensis y Candida albicans. Se utilizaron 6 cepas control de C. dubliniensis y una de C. albicans, provenientes de colecciones reconocidas y sometidas a genotipificación. También se utilizaron 70 aislados identificados como posibles C. albicans utilizando el medio CHROMagar Candida y el medio de bilis agar Feo. Los métodos evaluados fueron: agar Sabouraud dextrosa a 45°C, agar Sabouraud con NaCl al 6,5%, agar Tween 80, agar tabaco, agar Pal’s, agar tomate-zanahoria y aglutinación con partículas de látex (Bichro-Dubli Fumouze®). Encontramos que las técnicas más confiables para realizar la diferenciación fenotípica entre estas dos especies fueron: el agar tomate-zanahoria, el agar Pal’s, el agar tabaco y la aglutinación con partículas de látex (Bichro-Dubli Fumouze®). Además en este estudio, de los 70 aislados considerados como C. albicans, encontramos 1 (1.4%) posible Candida dubliniensis. Sin embargo, las pruebas de biología molecular son las más adecuadas para el diagnóstico certero de estas dos especies.

Candida albicans and Candida dubliniensis have a close phylogenetic relationship. The similarity between these species can cause a microbiology laboratory to identify C. dubliniensis erroneously as C. albicans. The objective of this research was to evaluate diverse phenotypic methods for differentiating between Candida dubliniensis and Candida albicans. Six control strains of C. dubliniensis and one of C. albicans from recognized collections were used and submitted to genotypification. Also, 70 isolates were used, identified as possible C. albicans utilizing CHROMagar Candida and bilis agar Feo mediums. The methods evaluated were: Sabouraud dextrosa agar at 45°C, Sabouraud agar with NaCl at 6.5%, Tween 80 agar, tobacco agar, Pal’s agar, tomato-carrot agar and agglutination with latex particles (Bichro-Dubli Fumouze®). It was found that the most reliable techniques for performing phenotype differentiation between these two species were tomato-carrot agar, Pal’s agar, tobacco agar and agglutination with latex particles (Bichro-Dubli Fumouze®). Of the 70 isolates considered to be C. albicans, one (1.4%) possible Candida dubliniensis was found. Nevertheless, molecular biology tests are the most appropriate means for achieving an accurate diagnosis of these two species.

Daptomycin-oxacillin combinations in treatment of experimental endocarditis caused by daptomycin-nonsusceptible strains of methicillin-resistant Staphylococcus aureus with evolving oxacillin susceptibility (the "seesaw effect").

In vivo development of daptomycin resistance (DAPr) among Staphylococcus aureus strains, especially methicillin-resistant S. aureus (MRSA) strains, in conjunction with clinical treatment failures, has emerged as a major problem. This has raised the question of DAP-based combination regimens to enhance efficacy against such strains. We studied five recent DAP-susceptible (DAPs)/DAPr clinical MRSA strain pairs obtained from patients who failed DAP monotherapy regimens, as well as one DAPs/DAPr MRSA strain pair in which the resistant strain was generated by in vitro passage in DAP. Of note, we identified a DAP-oxacillin (OX) "seesaw" phenomenon in vitro in which development of DAPr was accompanied by a concomitant fall in OX resistance, as demonstrated by 3- to 4-fold decreases in the OX MIC, a susceptibility shift by population analyses, and enhanced early killing by OX in time-kill assays. In addition, the combination of DAP and OX exerted modest improvement in in vitro bactericidal effects. Using an experimental model of infective endocarditis and two DAPs/DAPr strain pairs, we demonstrated that (i) OX monotherapy was ineffective at clearing DAPr strains from any target tissue in this model (heart valve, kidneys, or spleen) and (ii) DAP-OX combination therapy was highly effective in DAPr strain clearances from these organs. The mechanism(s) of the seesaw effect remains to be defined but does not appear to involve excision of the staphylococcal cassette chromosome mec (SCCmec) that carries mecA.

Impregnation of bone chips with antibiotics and storage of antibiotics at different temperatures: an in vitro study.

BACKGROUND: Allograft bone used in joint replacement surgery can additionally serve as a carrier for antibiotics and serve as a prophylaxis against infections. However, in vitro dose-response curves for bone chips impregnated with different kinds of antibiotics are not available. In addition, while it would be desirable to add the antibiotics to allograft bone chips before these are stored in a bone bank, the effects of different storage temperatures on antibiotics are unknown. METHODS: Five different antibiotics (cefazolin, clindamycin, linezolid, oxacillin, vancomycin) were stored, both as pills and as solutions, at -80 degrees C, -20 degrees C, 4 degrees C, 20 degrees C and 37 degrees C; in addition, bone chips impregnated with cefazolin and vancomycin were stored at -80 degrees C and -20 degrees C. After 1 month, 6 months and 1 year, the activity of the antibiotics against Staphylococcus epidermidis was measured using an inoculated agar. The diameter of the S. epidermidis-free zone was taken as a measure of antibiotic activity. In a separate experiment, in vitro dose-response curves were established for bone chips impregnated with cefazolin and vancomycin solutions at five different concentrations. Finally, the maximum absorbed amounts of cefazolin and vancomycin were established by impregnating 1 g of bone chips with 5 ml of antibiotic solution. RESULTS: A decrease of the S. epidermidis-free zone was seen with oxacillin and cefazolin solutions stored at 37 degrees C for 1 month, with vancomycin stored at 37 degrees C for 6 months and with cefazolin and oxacillin solutions stored at 20 degrees C for 6 months. The activity of the other antibiotic solutions, pills and impregnated bone chips was not affected by storage. The in vitro dose-response curves show that the free-zone diameter increases logarithmically with antibiotic concentration. The absorbed antibiotic amount of one gram bone chips was determined. CONCLUSIONS: Storage of antibiotics in frozen form or storage of antibiotic pills at temperatures up to 37 degrees C for 12 months does not affect their activity. However, storage of antibiotic solutions at temperatures above 20 degrees C does affect the activity of some of the antibiotics investigated. The in vitro dose-response curve can be used to determine the optimal concentration(s) for local application. It provides the opportunity to determine the antibiotic content of bone chips, and thus the amount of antibiotics available locally after application.

Species distribution and antimicrobial susceptibility of staphylococci isolated from canine otitis externa.

The diversity of species of the genus Staphylococcus sp. and the antimicrobial resistance of isolates from 151 unmedicated dogs of both sexes with a clinical diagnosis of otitis were recorded. Ninety-one isolates of Staphylococcus spp. were identified by biochemical reactions and tested for susceptibility to 15 antimicrobials. Coagulase-positive species were most common; S. pseudintermedius (38.4%), S. schleiferi schleiferi (15.4%), S. aureus (14.3%), S. epidermidis (11%), S. simulans (11%), S. schleiferi coagulans (8.8%) and S. saprophyticus (1.1%). All the isolates showed resistance to at least one drug and 89% were multiresistant. Amoxicillin combined with clavulanic acid and oxacillin were the most effective, while resistance was widely observed for neomycin and erythromycin. The results highlight the recognition and the potential need for bacterial culture with species identification and antimicrobial susceptibility tests for appropriate antimicrobial therapy.

Avaliação da colonização nasal por Staphylococcus spp. resistenteà oxacilina em alunos de enfermagem / Evaluation of nasal colonization for oxacillin resistant Staphylococcus spp. in nursing students

INTRODUÇÃO: O gênero Staphylococcus é responsável por um grande número de infecções bacterianas em humanos, principalmente no ambiente hospitalar. OBJETIVO: Diante dessas considerações, da importância do cuidado de enfermagem e do controle de infecções hospitalares, este trabalho verificou a taxa de portadores de S. aureus e estafilococos coagulase-negativa (ECN) resistentes a oxacilina em alunos do curso de enfermagem durante a formação profissional. MATERIAS E METÓDOS: Foram coletadas amostras das fossas nasais de alunos do curso de enfermagem da Faculdade de Medicina de Botucatu (FMB). RESULTADOS: Das 109 amostras de Staphylococcus encontradas, 30 (27,5 por cento) foram de Staphylococcus aureus e 79 (72,5 por cento), de ECN. Das 79 amostras de ECN submetidas às provas de identificação, 63 (79,7 por cento) foram da espécie S. epidermidis, nove (11,4 por cento), S. warneri, três (3,8 por cento), S. haemolyticus, duas (2,5 por cento), S. capitis, uma (1,3 por cento), S. simulans e uma (1,3 por cento) S. lugdunensis. O teste de suscetibilidade a antibióticos demonstrou 100 por cento de sensibilidade às drogas nas amostras de S. aureus e, entre as 79 amostras de ECN, 10 (12,6 por cento) foram resistentes a oxacilina. A técnica de reação em cadeia da polimerase (PCR) demonstrou resultado negativo para o gene mecA nas amostras de S. aureus e 11 amostras positivas entre as espécies de ECN. DISCUSSÃO: Não houve relação entre a taxa de portadores de S. aureus e o envolvimento hospitalar durante a graduação, os resultados também demonstraram maior incidência de resistência nas amostras de ECN, semelhante ao encontrado na literatura científica.

INTRODUCTION: The genus Staphylococcus is responsible for a great number of bacterial infections in human, mainly in hospital environment. OBJECTIVES: In view of these considerations and the importance of nursing care and nosocomial infection control, this study verified the rate of S. aureus and coagulase-negative Staphylococci (CNS) carriers resistant to oxacillin in nursing students during their university course. MATERIALS AND METHODS: Nasal samples were collected from Nursing students at Botucatu School of Medicine - UNESP. RESULTS: From 109 isolated samples of Staphylococcus, 30 samples (27.5 percent) were Staphylococcus aureus and 79 samples (72.5 percent) were CNS. From 79 identified CNS samples, 63 (79.7 percent) were S. epidermidis, nine (11.4 percent) S. warneri, three (3.8 percent) S. haemolyticus, two (2.5 percent) S. capitis, one (1.3 percent) S. simulans and one (1.3 percent) S. lugdunensis. The antibiotic susceptibility test showed 100 percent sensibility to the drugs in S. aureus samples and among 79 CNS samples, 10 (12,6 percent) were resistant to oxacillin. The PCR technique demonstrated negative result for mecA gene in S. aureus samples and 11 positive samples among CNS species. DISCUSSION: There was no relation between the rate of S. aureus carriers and nosocomial involvement during the course. The results also showed a higher incidence of resistance in CNS samples, which is seemingly reported in the scientific literature.

Methicillin and aminoglycoside resistance in Staphylococcus aureus isolates from bovine mastitis and sequence analysis of their mecA genes.

Although methicillin-resistant Staphylococcus aureus (MRSA) were generally isolated from human beings; these agents were recently isolated from various animal species. It has been shown that MRSA isolates are not only resistant to beta-lactam antibiotics, but can also be resistant to the other commonly used antibiotics. In this study, 18 phenotypic methicillin resistant S. aureus isolates from bovine mastitis cases were analyzed by PCR for the presence of mecA gene encoding methicillin resistance and aac (6')/aph(2″), aph(3')-IIIa and ant(4')-Ia genes encoding aminoglycoside resistance. Out of 18 S. aureus isolates (oxacillin MICs, ≥4 µg/ml), 3 were positive for mecA gene. Only one from 3 mecA positive isolates was positive for genes encoding aminoglycoside-modifying enzymes and this isolate carried aac(6')/aph(2″) in combination with aph(3')-IIIa gene. The aph(3')-IIIa gene was detected in 3 isolates. These three isolates carrying the aminoglycoside-modifying enzyme genes were resistant to gentamicin, kanamycin and neomycin. The mecA gene of 3 MRSA isolates was sequenced. All three mecA genes of these isolates were identical to that found in human MRSA strains, except a one-base substitution at nucleotide position 757. From the data presented in this study, it can be concluded that MRSA isolated from bovine mastitis may be originated from human beings, but further studies are needed to investigate the possibility of zoonotic transfer of MRSA.

In vitro activity of daptomycin against Staphylococci isolated from bacteremia and community-onset skin and soft tissue infections in France: data from two nationwide studies.

Staphylococci are a leading cause of skin and soft tissue infections (SSTIs) and bacteremia in France, a country with a high prevalence of oxacillin resistance. We evaluated the in vitro activity of daptomycin compared with reference compounds against 445 Staphylococcus aureus and 53 coagulase-negative Staphylococci (CNS) collected during two large nationwide studies performed in 2006 and 2007. The percentage of oxacillin resistance among S. aureus was 13.6% (SSTIs) and 30.7% (bacteremia). Daptomycin showed lower MIC(90) levels compared to vancomycin, teicoplanin, and linezolid (0.19 mg/L vs. 2, 1.5, and 1 mg/L, respectively), irrespective of oxacillin susceptibility. Amongst the CNS, 64.2% of the isolates originated from clinical bacteremia were resistant to oxacillin and 24.5% to teicoplanin; all but one Staphylococci were susceptible to daptomycin (MIC = 1.5 mg/l). As with linezolid, daptomycin seems to constitute an alternative option to treat some staphylococcal infections in the French context of high oxacillin resistance prevalence and high glycopeptides MIC.