Activity of combinations of ceftazidime, imipenem and pefloxacin against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa

The chequerboard technique was used to look for synergistic combinations of ceftazidime, imipenem and pefloxacin. The synergistic combinations were used in vivo in mice experimentally infected with Escherichia coli, Salmonella typhimurium and Pseudomonas aeruginosa. In vitro ceftazidime/imipenem, ceftazidime/pefloxacin and pefloxacin/imipenem combinations showed synergistic effects against Staphylococcus aureus and S. typhimurium and additive effects against P. aeruginosa. Only the ceftazidime/pefloxacin combination was synergistic against E. coli while the ceftazidime/imipenem and pefloxacin/imipenem combinations resulted in additive effects. In vivo, combination of ceftazidime/imipenem against E. coli infection and the pefloxacin/imipenem combination against S. typhimurium infection were protective.     

Synthesis and evaluation of lysozyme derivatives exhibiting an enhanced antimicrobial action

In order to generate novel preservatives exhibiting a broad antimicrobial spectrum against Gram-positive as well as Gram-negative bacteria, lysozyme was modified by the covalent attachment of caffeic acid and cinnamic acid, respectively. Linkage of these organic acids to lysozyme was achieved by the constitution of amide bindings between the carboxyl group of ligands and primary amino groups of the enzyme mediated by a carbodiimide. Compared to nonmodified lysozyme, the lytic activity of all resulting conjugates was reduced. In contrast, bacterial growth of Escherichia coli (ATCC 8739) could be strongly inhibited by lysozyme–caffeic acid conjugates and to a lower degree also by lysozyme–cinnamic acid conjugates. The minimal inhibitory concentration against E. coli was 0.05% for the lysozyme derivative of the highest antimicrobial activity. However, the efficacy of lysozyme derivatives against Staphylococcus aureus (ATCC 6538) was slightly reduced. As the antimicrobial spectrum of lysozyme altogether could be substantially widened, these derivatives represent promising candidates as novel preservatives for various pharmaceutical and cosmetic formulations.     

Activity of tigecycline against clinical isolates of Staphylococcus aureus and extended-spectrum beta-lactamase-producing Escherichia coli in Granada, Spain

We evaluated the in vitro activity of tigecycline using the Etest and disk diffusion method according to Clinical and Laboratory Standards Institute guidelines against clinical isolates of methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) as well as for CTX-M-9 extended-spectrum β-lactamase (ESBL)-producing Escherichia coli and SHV ESBL-producing E. coli. All isolates were susceptible to tigecycline according to US Food and Drug Administration cut-off points. There were no differences in the activity of tigecycline between MSSA and MRSA isolates or between the presence of either type of ESBL. For each type of microorganism studied, we established the equation relating the minimum inhibitory concentration to the diameter of the zone of inhibition.     

Alcohol ethoxylates mediate their bacteriostatic effect by altering the cell membrane of Escherichia coli NCTC 8196

The minimum inhibitory concentration (MIC) of a homologous series of alcohol ethoxylates with the same head group size (E6) but differing in the number of carbon atoms in their ‘tail group’ from 10 to 16 was determined for Staphylococcus aureus NCTC 4163 and Escherichia coli NCTC 8196 using a turbidimetric assay. All the surfactants tested demonstrated bacteriostatic activity against both organisms. A tetrazolium assay showed that C14E6 and C16E6 had little effect on the membrane-bound dehydrogenase enzyme activity of E. coli NCTC 8196 compared with C10E6 and C12E6. C10E6 caused leakage both of K⁺ and nucleotides in a concentration-dependent manner above its MIC of 0.2 mM. C12E6 caused some leakage at concentrations below its MIC (0.12 mM).     

In vitro activity of lauric acid or myristylamine in combination with six antimicrobial agents against methicillin-resistant Staphylococcus aureus (MRSA)

The objective of this study was to investigate the in vitro activities of lauric acid and myristylamine in combination with six antimicrobial agents against methicillin-resistant Staphylococcus aureus (MRSA). The combination effect of lipids and antimicrobial agents was evaluated by the checkerboard method to obtain a fractional inhibitory concentration (FIC) index. The effects of lauric acid + gentamicin (GM) and lauric acid + imipenem (IPM) combinations were synergistic against the clinical isolates in 12 combinations. An antagonistic FIC index was observed only with the myristylamine + GM combination. We investigated in detail the antimicrobial activity for two combinations that showed a synergistic effect. The cytotoxicity of lauric acid was not enhanced by the addition of GM and IPM. In time-kill studies, lauric acid + GM and lauric acid + IPM combinations at one-eighth of the minimum inhibitory concentration produced a bacteriostatic effect.     

Age-related trends in pathogen frequency and antimicrobial susceptibility of bloodstream isolates in North America: SENTRY Antimicrobial Surveillance Program, 1997-2000

We report age-related trends in pathogen frequency and antimicrobial susceptibility from 25,745 bloodstream infections (BSI) due to bacterial pathogens reported from medical centres participating in the North American SENTRY Antimicrobial Surveillance Program between January 1997 and September 2000. Staphylococcus aureus, Escherichia coli and coagulase-negative staphylococci (CoNS) were the most common pathogens, together accounting for 55% of all BSI pathogens during this time period. Among nosocomial BSI, CoNS were the most frequently isolated pathogens in infants less than 1 year of age, but S. aureus increased in frequency with increasing age. Among community-onset BSI pathogens, Streptococcus pneumoniae was the most frequently reported pathogen causing BSI in patients aged 1–5, S. aureus among those aged 6–64, and E. coli predominated at the extremes of age (less than 1 year and ≥65 years of age). Among key organism: antimicrobial agent combinations evaluated, oxacillin resistance in S. aureus increased with increasing age; conversely, oxacillin resistance among CoNS was highest among children 5 years of age or younger. Penicillin resistance among S. pneumoniae BSI was highest in children younger than 5 years, while vancomycin resistance among Enterococcus spp. predominated among nosocomial BSI in patients over 50 years of age. Important age-related differences exist in species distribution and antimicrobial susceptibility of pathogens causing BSI. This information should be helpful for clinicians as they consider empirical antimicrobial therapy for patients with suspected BSI across the age continuum.     

Antimicrobial resistance in Gram-negative bacteria from Intensive Care Units and Urology Services. A nationwide study in The Netherlands 1995-2000

A nationwide 6-year surveillance of resistance in Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis and Pseudomonas aeruginosa from clinical specimens of patients from ten Intensive Care Units and ten Urology Services was started in 1995. MICs of amoxycillin, amoxycillin/clavulanate, trimethoprim, cotrimoxazole, norfloxacin, ciprofloxacin, cefaclor, ceftazidime, imipenem and gentamicin were determined by broth microdilution. Intensive Care Units had higher resistance levels of amoxycillin/clavulanate, cefaclor and ceftazidime (P<0.005) and lower resistance levels of nitrofurantoin, trimethoprim, cotrimoxazole and quinolones (P<0.01) than Urology Services. Changes in MIC distributions in time and development of resistant clusters were observed for nitrofurantoin (E. coli), amoxycillin (E. coli, P. mirabilis), amoxycillin/clavulanate (E. coli) and for quinolones (E. coli). The overall resistance level of ceftazidime and gentamicin was <5%, but this fluctuated with the appearance and disappearance of resistant clones in some Intensive Care Units. Quinolone resistance among P. aeruginosa from Intensive Care Units fluctuated between 7 and 14%.     

Influence of the cell wall on ciprofloxacin susceptibility in selected wild-type Gram-negative and Gram-positive bacteria

The susceptibility of several wild-type bacteria to ciprofloxacin and accumulation of the drug in these bacteria were evaluated. Species studied included Escherichia coli, Serratia marcescens, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Bacillus cereus. Ciprofloxacin susceptibility was measured for each strain using two different methods: the minimal inhibitory concentration and the bactericidal index. Significant differences were observed between the results derived from these two methods. Whereas the minimal inhibitory concentration was low in all strains tested, ciprofloxacin’s bactericidal activity, as indicated by the bactericidal index, varied with the species studied. To determine whether this finding was due to variations in cell envelope permeability to ciprofloxacin (i.e. to combined cell uptake and efflux), we studied ciprofloxacin accumulation using spectrofluorometry. In Gram-negative bacteria, differences in permeability can lead to altered susceptibility to antibiotics. In fact, the combination of slow uptake and efficient efflux seems to be crucial to the characteristic poor susceptibility of P. aeruginosa to ciprofloxacin. However, the low level of activity of ciprofloxacin against S. aureus and two Bacillus species may have resulted from the drug’s interaction with its target enzymes (i.e. topoisomerase IV in S. aureus and DNA gyrase in Bacillus spp.) rather than diminished permeability.     

Extended antimicrobial resistance screening of the dominant faecal Escherichia coli and of rare resistant clones

Fifty faecal samples from healthy adults were grown on MacConkey agar and three pink colonies were subcultured, identified to species level and their antimicrobial susceptibility determined. Forty-seven samples yielded 141 isolates of Escherichia coli that were susceptible to most antimicrobials. Resistance was noted for ampicillin (30.5%), chloramphenicol (12.1%), tetracycline (23.4%), trimethoprim (24.8%) and co-trimoxazole (22.7%). A direct faecal plating method was used for extended resistance screening with E. coli as the indicator organism. Zone breakpoints were determined using normalised resistance interpretation and gave similar susceptibility results. Eighty-eight isolates of E. coli from within the zones of inhibition revealed four times more antimicrobial resistance. Extended antimicrobial resistance screening both provides the susceptibility profile of the dominant E. coli isolate and detects greater resistance in rare isolates.     

Relationships between antimicrobial use and antimicrobial resistance in Gram-negative bacteria causing nosocomial infections from 1991-2003 at a university hospital in Taiwan

This study was conducted to evaluate the relationship between antimicrobial resistance and antimicrobial use in a university hospital in Taiwan. Disk susceptibility data of Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Serratia marcescens, Proteus spp., Pseudomonas aeruginosa, Acinetobacter spp., Stenotrophomonas maltophilia and other non-fermentative Gram-negative bacilli causing nosocomial infections were evaluated. Data on annual patient-days and annual consumption (defined daily dose (DDD) per 1000 patient-days) of extended-spectrum cephalosporins (cefotaxime, ceftriaxone, ceftazidime, flumoxef, cefepime and cefpirome), β-lactam–β-lactamase inhibitor combinations (ticarcillin/clavulanic acid and piperacillin/tazobactam), carbapenems (imipenem and meropenem), aminoglycosides (amikacin, gentamicin and tobramycin), fluoroquinolones (ciprofloxacin (oral and injectable) and oral levofloxacin and moxifloxacin) from 1991 to 2003 were analysed. Increasing trends of incidences of several of these bacteria causing all nosocomial infections or nosocomial bloodstream infections were noted from 1991 to 2003. The annual patient-days of the hospital significantly increased, from 360 210 in 1991 to 672 676 in 2002 (linear regression analysis, P < 0.05), but slightly decreased in 2003 (629 168) owing to the severe acute respiratory syndrome epidemic in Taiwan. The rise in cefotaxime-resistant or ciprofloxacin-resistant E. coli and meropenem-resistant P. aeruginosa was significantly correlated with increased consumption of extended-spectrum cephalosporins, β-lactam–β-lactamase inhibitor combinations, carbapenems, fluoroquinolones and aminoglycosides (for ciprofloxacin-resistant E. coli and meropenem-resistant P. aeruginosa only) in the hospital (Pearson's correlation coefficient, r > 0.72 (or <−0.72) and P-value < 0.05). Increased ciprofloxacin-resistant K. pneumoniae and meropenem-resistant Acinetobacter spp. was significantly associated with the increased usage of extended-spectrum cephalosporins but not with the other four classes of antibiotics. This 13-year study in a hospital demonstrated significant changes in antimicrobial use, which may have affected antimicrobial resistance in certain Gram-negative bacteria at the hospital.     

噁唑烷酮-地拉罗司偶联物的合成及抗菌活性评价（英文）

本文基于“特洛伊木马”策略设计合成了两种具有不同连接臂的噁唑烷酮-地拉罗司偶联物,并进行了抗菌活性评价。与地拉罗司相同,两种偶联物均可与Fe³⁺结合。然而,其对所测试的菌株(包括金黄色葡萄球菌、大肠杆菌、鲍曼不动杆菌和铜绿假单胞菌)均无活性。结果表明,合成的铁螯合剂地拉罗司可能不适合作为细菌转运抗生素的铁离子载体,或者合成的偶联物的连接臂不能在细菌细胞质中水解释放噁唑烷酮,噁唑烷酮-地拉罗司偶联物的设计与合成还需要进一步探索。     

Antimicrobial resistance of bacterial isolates from respiratory care wards in Taiwan: a horizontal surveillance study

A horizontal surveillance study was conducted to identify common bacteria and mycobacteria from 611 respiratory aspirates and 165 urinary samples from 611 patients hospitalised at 17 respiratory care wards (RCWs) in Taiwan. Some major resistance phenotypes, including meticillin-resistant Staphylococcus aureus (MRSA), extended-spectrum β-lactamase (ESBL)-producing Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis, and multidrug-resistant Pseudomonas aeruginosa (MDR-PA) and Acinetobacter baumannii (MDR-AB), were identified. Pulsotypes of ESBL-producing P. mirabilis isolates were determined by pulsed-field gel electrophoresis. The prevalences of MRSA, ESBL-producing E. coli (K. pneumoniae and P. mirabilis), carbapenem-resistant (resistant to imipenem and meropenem) P. aeruginosa, MDR-PA, carbapenem-resistant A. baumannii and MDR-AB were, respectively, 86.7%, 20.0% (50.7% and 24.1%), 18.4%, 1.2%, 32.1% and 8.9% for respiratory aspirates and 100%, 25.4% (27.3% and 25.0%), 48.3%, 10.3%, 50.0% and 21.4% for catheterised urinary samples. Among the 44 respiratory isolates of P. mirabilis with an ESBL phenotype, 22 different pulsotypes (>80% identity) were identified. Among 103 isolates of mycobacteria, 90 (87.4%) belonged to rapidly growing mycobacteria and 4 (4%) were Mycobacterium tuberculosis. Among the 404 patients with available clinical information, true infections were found in 28.0%, the most prevalent of which were urinary tract infection (20.5%) and ventilator-associated pneumonia (10.9%). High prevalences of various multidrug-resistant bacteria among the respiratory and urinary tracts of patients present a clinical difficulty in choosing empirical antibiotic treatment in RCWs.     

Antimicrobial activity of imipenem against isolates from complicated urinary tract infections

Antimicrobial activity of imipenem was measured using 4725 strains isolated from patients with complicated urinary tract infections (CUTIs) between 1988 and 2000. Imipenem was inactive against methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis, Enterococcus faecium and some non-fermenting Gram-negative rods. Resistant strains (MIC>16 mg/l) were observed in Staphylococcus haemolyticus (22%), Enterococcus faecalis (4%), Enterococcus avium (8%), Serratia marcescens (5%) and Pseudomonas aeruginosa (7%). Although the prevalence of imipenem-resistant strains of S. aureus, S. epidermidis and P. aeruginosa was sporadically high in some years, no steady increase was seen over the period. Resistant strains were rare in other major uropathogenic species. These results suggest that imipenem is still one of the most reliable antimicrobial drugs.     

A seven-year survey of susceptibility to marbofloxacin of pathogenic strains isolated from pets

This study, Vétoquinol S.A. epidemiosurveillance, was conducted from 1994 to 2001 in order to determine the susceptibility (by MIC determination) to marbofloxacin (a third generation fluoroquinolone used only in individual administration for animals). Strains from infected pets originated from six European countries. Isolates were collected from urinary infections (Escherichia coli), respiratory infections (Pasteurella multocida), dermatological infections (Staphylococcus intermedius, Pseudomonas aeruginosa) and otitis (S. intermedius, P. aeruginosa). The MIC distribution for each species was the same both before and after the launch of marbofloxacin in 1995. In E. coli, a resistant population was present before the use of marbofloxacin; this resistance was induced by co- or cross-resistance to other antibiotics used previously. Over this period, there was no significant evolution of MIC₉₀ for any bacterial species studied and no development of resistance was observed. Marbofloxacin was the most active antibiotic against P. multocida isolates and had the lowest MIC. No difference in MIC distribution was seen between the S. intermedius (unimodal distribution) isolated from dermatological infections and those from otitis. This was also true for P. aeruginosa. The use of marbofloxacin was not found to have induced a significant increase or spread of resistant bacteria.     

木犀草素降低大肠埃希菌对呼吸道上皮细胞感染的作用机制研究

目的：探讨亚抑菌浓度（sub-minimal inhibitory concentration,亚-MIC）木犀草素降低临床分离大肠埃希菌对呼吸道上皮细胞HBE感染的作用机制,为临床抗感染的治疗提供新的思路。方法：采用微量肉汤稀释法检测木犀草素对大肠埃希菌临床株E367的MIC值,采用多功能酶标仪测定亚-MIC木犀草素对细菌生长的影响,采用CCK-8法测定亚-MIC木犀草素处理大肠埃希菌对HBE细胞毒性的影响,采用ELISA试剂盒测定细胞炎症因子白细胞介素（interleukin,IL）-6、IL-8及肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）水平,采用Western Blot测定p-p65/p65、p-IκBα/IκBα蛋白表达水平。结果：1/4 MIC-1/32 MIC木犀草素对大肠埃希菌的生长没有影响,且能剂量依赖性地降低大肠埃希菌对呼吸道上皮细胞的毒性。大肠埃希菌感染细胞后其炎症因子IL-6、IL-8、TNF-α水平和p-p65/p65、p-IκBα/IκBα蛋白表达水平均显著增高。与细菌处理组相比,木犀草素和大肠埃希菌共同处理细胞后其IL-6、TNF-α水平及p-p65/p65、p-IκBα/IκBα蛋白表达水平显著降低,而细胞因子IL-8水平几乎不变。结论：木犀草素可能通过免疫调节抑制大肠埃希菌对呼吸道上皮细胞的感染作用。     

Resistance of bacteria in urinary tract infections

Bacterial infection of the urinary tract is a common health problem in young women but also the most common nosocomial infection (>33%) contributing to the mortality of patients, and increasing the duration and cost of hospitalization. Escherichia coli is the most predominant organism and its prevalence varies in different studies. The high consumption of inappropriately prescribed antibiotics, combined with multiple pathology and frequent use of invasive devices, is a major factor contributing to high levels of resistance. There is a serious decrease in susceptibility of E. coli strains to amoxycillin, due to the presence of R-TEM enzymes, to cotrimoxazole and trimethoprim. Nitrofurantoin and fosfomycin-trometamol remain highly active against urinary Enterobacteriaceae, with over 90% of E. coli being susceptible. Knowledge of the most likely causative organisms and the prevalence of resistance pathogens to antimicrobial agents is essential to select antibiotics and to establish guidelines for the empirical treatment of urinary tract infections.     

Target site bacterial killing of cefpirome and fosfomycin in critically ill patients

We employed an in-vivo pharmacokinetic/in-vitro pharmacodynamic method to simulate bacterial killing in plasma and the interstitium of skeletal muscle tissue after intravenous administration of 2 g of cefpirome and 8 g of fosfomycin alone and in combination to patients with sepsis. Interstitial antimicrobial concentrations were determined by use of in-vivo microdialysis. CFU/ml of Staphylococcus aureus (ATCC 29213) and Pseudomonas aeruginosa (clinical isolate) decreased by approximately 2 log₁₀ for plasma and muscle tissue 6 h after cefpirome and fosfomycin administration compared with the baseline, respectively. The simulation of plasma and tissue pharmacokinetics for the combined administration of these antibiotics resulted in complete eradication of S. aureus within 5 h after drug exposure. No bacterial re-growth occurred in any of the simulations within 6 h. The in-vitro simulation of in-vivo plasma and tissue pharmacokinetics of cefpirome and fosfomycin has shown that both antimicrobial agents kill S. aureus and P. aeruginosa strains effectively after single dose administration. This effect was most pronounced by the combined use of these antimicrobial agents. Therefore, this data corroborates antimicrobial strategies of simultaneous administration of cefpirome and fosfomycin in patients with severe soft tissue infection.     

Bioactive properties and chemical composition of six walnut (Juglans regia L.) cultivars

The chemical composition, antioxidant potential and antimicrobial activity were studied in six walnuts (Juglans regia L.) cultivars (cv. Franquette, Lara, Marbot, Mayette, Mellanaise and Parisienne) produced in Portugal. Concerning their chemical composition the main constituent of fruits was fat ranging from 78.83% to 82.14%, being the nutritional value around 720 kcal per 100 g of fruits. Linoleic acid was the major fatty acid reaching the maximum value of 60.30% (cv. Lara) followed by oleic, linolenic and palmitic acids. The aqueous extracts of walnut cultivars were investigated by the reducing power assay, the scavenging effect on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and β-carotene linoleate model system. All the walnut extracts exhibited antioxidant capacity in a concentration-dependent manner being the lowest EC₅₀ values obtained with extracts of cv. Parisienne. Their antimicrobial capacity was also checked against gram positive (Bacillus cereus, Bacillus subtilis, Staphylococcus aureus) and gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae) and fungi (Candida albicans, Cryptococcus neoformans), revealing activity against the different tested microorganisms.     

The effect of Nepeta cataria extract on adherence and enzyme production of Staphylococcus aureus.

Nepeta cataria L., commonly known as catnip, is a perennial herb with a considerable folkloric reputation. A diethyl ether extract of this plant has been shown to have antimicrobial activity against fungi and Gram-positive bacteria. The aim of this work was to study the activity of N. cataria extract on 44 Staphylococcus aureus strains, some resistant to methicillin, and S. aureus 6538P (American Type Culture Collection) by evaluating the effect of subminimum inhibitory concentrations on coagulase, DNAse, thermonuclease and lipase production, and on in-vitro adherence. DNAse, thermonuclease and lipase were inhibited by concentrations equal to 1/2 and 1/4 MIC. A reduction of adherence was also observed.     

Enhancement of the antibacterial activity of ampicillin by liposome encapsulation

This study showed that encapsulation of ampicillin (Amp) in liposomes prepared with synthetic lecithins enhanced its antibiotic activity against both Amp-sensitive and Amp-resistant Escherichia coli. This was demonstrated by growth inhibition of E. coli in the presence of the liposomal preparations containing Amp. Growth inhibition was also seen in the presence of exogenous β-lactamase. Adsorption of Amp onto the surface of the liposomes did not result in enhanced activity of the drug against E. coli. The encapsulation efficiency of Amp in liposomes prepared by the Bangham method (BM) was greatly affected by the phospholipids used. The efficiency increased with a rise in the phase transition temperature (T_c) of the phospholipid, the maximum encapsulation of Amp being obtained with distearoylphosphatidylcholine (DSPC). The entrapment efficiency of Amp with the reverse phase evaporation method (REV) was largely superior to that with the BM method. Kinetic studies using DSPC liposomes prepared by the REV and BM protocols demonstrated that Amp-loaded liposomes contained 60 and 72% of drug, respectively, after 24 h at 37°C.