泵抑制剂对嗜麦芽寡养单胞菌临床分离株氟喹诺酮类敏感性的影响

目的 了解嗜麦芽寡养单胞菌临床分离株外排泵的分布以及不同泵抑制剂对该菌氟喹诺酮类敏感性的影响。方法 用琼脂二倍稀释法检测泵抑制剂存在时，嗜麦芽寡养单胞菌对环丙沙星、氧氟沙星和诺氟沙星的敏感性变化。结果 羰基氰氯苯腙(CCCP)和利舍平降低部分嗜麦芽寡养单胞菌株对氟喹诺酮的耐药性。维拉帕米无明显泵抑制作用。泵阳性株存在于耐药和非耐药菌株中，集中于耐药性较高的菌株。泵抑制剂对该菌外排氧氟沙星的抑制大于对诺氟沙星和环丙沙星的作用。结论 多重外排泵是嗜麦芽寡养单胞菌对氟喹诺酮类耐药的原因之一。泵抑制剂可部分逆转这种耐药性。     

Comparative activities of six different fluoroquinolones against 9,682 clinical bacterial isolates from 20 European university hospitals participating in the European SENTRY surveillance programme

The in-vitro activities of gatifloxacin, trovafloxacin, levofloxacin, sparfloxacin, ofloxacin, and ciprofloxacin were tested against 9,682 clinical bacterial isolates from 20 European university hospitals participating in the European SENTRY surveillance programme. Gatifloxacin and trovafloxacin exhibited the highest activities against Gram-positive cocci, while levofloxacin, ofloxacin, ciprofloxacin, and gatifloxacin were the most active against Enterobacteriaceae. Ciprofloxacin and levofloxacin showed the highest antimicrobial activities against Pseudomonas spp., while gatifloxacin and trovafloxacin were the most active against Acinetobacter spp. and Stenotrophomonas maltophilia. All Haemophilus spp. and Moraxella catarrhalis isolates were fully susceptible to all quinolones tested. Overall, the new quinolones, showed improved activity against Gram-positive cocci and Gram-negative non-fermenters while retaining their broad-spectrum activity against Gram-negative bacilli.     

Rufloxacin (MF-934): in vitro and in vivo antibacterial activity

Rufloxacin (MF-934) is a new quinolone which shows in vitro antibacterial activity against E. coli, Salmonella, Klebsiella, Proteus and Staphylococcus spp. Lower in vitro activity was observed with Pseudomonas, Serratia, Enterobacter and the streptococci group D. The antimicrobial activity of MF-934 in vitro is higher than that of nalidixic acid but lower than that of ciprofloxacin, ofloxacin, pefloxacin or norfloxacin. The protective effects of MF-934 in systemic infections in mice are lower than those of ciprofloxacin and ofloxacin. In respiratory infections in rats and in subcutaneous infections in guinea-pigs, the protective effects of MF-934 are of the same order as ciprofloxacin and ofloxacin. This is probably due to the pharmacokinetic properties of MF-934, i.e. the long half-life and tissue concentrations higher than plasma levels.     

司帕沙星与其他5种抗菌药体外抗菌活性比较研究

测定了司帕沙星对临床分离的１９９ 株致病菌的体外抗菌活性并与氧氟沙星、环丙沙星、头孢唑啉、头孢噻肟、阿米卡星的抗菌活性进行比较。司帕沙星对革兰阳性菌的ＭＩＣ９０为０．１２５～０．５ｍ ｇ／Ｌ,对金葡球菌、化脓链球菌的抑菌率均为１００％ ,强于氧氟沙星、环丙沙星;对革兰阴性菌也具有良好的体外抗菌活性,与氧氟沙星、环丙沙星相似。不同细菌接种量和不同浓度血清对其抗菌活性无明显影响,仅在ｐＨ５．０ 时抗菌活性略有下降     

In vitro fluoroquinolone-resistant mutants of Salmonella enterica serotype Enteritidis: analysis of mechanisms involved in resistance

This study analysed the mechanisms involved in the acquisition of resistance to quinolones in mutants obtained in vitro of Salmonella enterica serotype Enteritidis. Two nalidixic acid-resistant (minimal inhibitory concentrations, MIC>256 mg/l), ciprofloxacin-susceptible (MIC 0.5 mg/l) clinical isolates of Salmonella Enteritidis with a mutation at amino acid codon Ser-83 of the gyrA gene were grown on plates containing increasing concentrations of ciprofloxacin. The increase in MIC to ciprofloxacin, sparfloxacin and trovafloxacin was totally or partially associated with over-expression of an AcrAB-like efflux pump. In addition, unidentified mechanism(s) may have been involved in the increased MIC to these antimicrobials. This study demonstrated that AcrAB-like efflux pumps appear to play a relevant role in the increase in MIC to some quinolones although, other, as yet undefined, mechanisms may be involved.     

多重耐药泵抑制剂提高肠球菌对氟喹诺酮药物敏感性的研究

目的　研究临床分离肠球菌对诺氟沙星、环丙沙星、氧氟沙星等氟喹诺酮药物耐药的主动泵出机制。方法　收集临床分离肠球菌 ;用琼脂二倍稀释法测定多重耐药泵抑制剂利舍平或维拉帕米应用前后 ,菌株对氟喹诺酮药物 MIC的变化。结果　利舍平使所有被检测的 2 9株肠球菌对诺氟沙星的敏感性增加 (MIC下降至原值的 1/ 2或以下 ) ,使 2 3株肠球菌对环丙沙星的敏感性增加 ,但仅能增加 3株肠球菌对氧氟沙星的敏感性。应用利舍平以后 ,12株耐诺氟沙星粪肠球菌对诺氟沙星 MIC值均下降 ,11株耐环丙沙星粪肠球菌有7株对环丙沙星 MIC值下降。维拉帕米的抑制效果和利舍平相比略有差异。结论　诺氟沙星、环丙沙星的主动泵出机制普遍存在于临床分离肠球菌 ,并且是粪肠球菌对诺氟沙星、环丙沙星的耐药机制之一。     

6种氟喹诺酮类药物对临床分离金葡萄的抗菌活性比较

目的：观察临床分离金葡萄对6种氟喹诺酮类药物的耐药情况。方法：用琼脂稀释法测定诺氟沙星（NFLX）、氧氟沙星（OFLX）、氟罗沙星（FLRX）、环丙沙星（CPFX）、司帕沙星（SPFX）、托舒沙星（TFLX）6种2、3代氟喹诺桐类药物对成都地区155株临床分离金葡菌的体外抗菌活性。结果：金葡菌对6种药物的耐药率分别为诺氟沙星35.5%。氟罗沙星34.19%,环丙沙星27.75%,托舒沙星27.75% ,氧氟沙星25.81%,司帕沙星25.81%。结论：氟喹诺酮类药物的广泛应用已使细菌的耐药性明显增高,各药MIC90均超过16mg.L^-1,比以往文献报道的本地区金葡萄耐药性明显升高,提示要合理应用氟喹诺酮类药物,减少耐药菌株的增加。     

In vitro susceptibilities of Bartonella and Rickettsia spp. to fluoroquinolone antibiotics as determined by immunofluorescent antibody analysis of infected Vero cell monolayers.

The in vitro susceptibilities of Bartonella and Rickettsia spp. to different concentrations of ciprofloxacin, levofloxacin, ofloxacin and sparfloxacin in Vero cell cultures, were determined by enumeration of immunofluorescent-stained bacilli. After incubation in a CO(2)-enriched atmosphere, inocula were replaced and tested with media containing 12 different concentrations of each antibiotic in replicate for each species and the monolayers were re-incubated. Growth status was determined by evaluation of immunofluorescent staining bacilli. Effective inhibitory antibiotic dilution endpoints were determined by counting Bartonella- and Rickettsia-specific fluorescent foci across a range of antibiotic dilutions with an epi-fluorescent microscope, and were compared with an antibiotic-negative control. Based upon the use of C(max):MIC and AUC:MIC data, levofloxacin exhibited activity against Bartonella elizabethae and B. quintana.     

Uptake of Fluoroquinolones in Human Monocytes Isolated from Peripheral Blood

The aim of this study was to develop a technique for separating monocytic cells in suspension from peripheral blood to measure the intracellular penetration of three fluoroquinolones (ofloxacin, ciprofloxacin and sparfloxacin). Mononucleated cells were isolated from the blood on a density gradient with lymphoprep and purified by a specific technique of adhesion and disadhesion on fibronectin. The monocytes were obtained in suspension with 76.8% purity and 97.9% viability. This was a convenient form for measurement of intracellular accumulation by use of the velocity-centrifugation technique. Intra-monocytic penetration of ciprofloxacin, ofloxacin and sparfloxacin was measured at equilibrium after 30-min incubation in the presence of 16 μg mL^?1 antibiotic. The results revealed low intra-monocytic accumulation of ciprofloxacin (intracellular-extracellular = 1.76) and ofloxacin (intracellular-extracellular = 1.42). The penetration of sparfloxacin was significantly higher (intracellular-extracellular = 2.4). This study confirms the important differences between human immunocompetent cells in terms of their ability to concentrate quinolones. It also underlines the importance of monocyte-macrophage cellular differentiation as a determinant of antibiotic penetration.     

Detection of novel gyrA mutations in nalidixic acid-resistant isolates of Salmonella enterica from patients with diarrhoea

The aim of the current study was to detect mutations in the gyrA gene of quinolone-resistant Salmonella spp. isolates recovered in Tehran, Iran. Between April 2008 and September 2009, 174 Salmonella spp. were collected and assayed for quinolone resistance and detection of gyrA mutations. Isolates identified as Salmonella enterica were tested for susceptibility by the disk diffusion method. Polymerase chain reaction (PCR) amplification and sequencing of the gyrA gene segment encoding the quinolone resistance-determining region (QRDR) were performed for the nalidixic acid-resistant isolates. Amongst the 174 recovered Salmonella spp. isolates, 89 were resistant to nalidixic acid, of which 9 were resistant to enrofloxacin; 10 isolates had reduced susceptibility to nalidixic acid. None of the isolates were resistant to ciprofloxacin, but a single isolate showed reduced susceptibility. Twelve types of amino acid replacement were found in the QRDR region of GyrA, namely the previously described substitutions in positions 83 and 87 as well as five new substitutions Leu41-Pro, Arg47-Ser, Ser111-Thr, Ala118-Thr and Asp147-Gly. Double substitutions in both positions 83 and 87 were not identified. A Gly133-Glu substitution was identified in a single S. enterica serotype Typhi isolate.     

Prolonged exposure of methicillin-resistant Staphylococcus aureus (MRSA) COL strain to increasing concentrations of oxacillin results in a multidrug-resistant phenotype

Our previous studies demonstrated that exposure of a bacterium to increasing concentrations of an antibiotic would increase resistance to that antibiotic as a consequence of activating efflux pumps. This study utilises the same approach; however, it employs the methicillin-resistant Staphylococcus aureus (MRSA) COL strain, which is highly resistant to oxacillin (OXA). MRSA COL was adapted to 3200 mg/L of OXA. Changes in resistance to other antibiotics were evaluated and efflux pump activity during the adaptation process was determined. MRSA COL was exposed to stepwise two-fold increases of OXA. At the end of each step, minimum inhibitory concentration determination for erythromycin (ERY) and other antibiotics was conducted. Reserpine (RES) was employed to evaluate whether resistance to ERY was dependent on efflux pump activity. Efflux pump activity was also evaluated using the ethidium bromide (EB) assay. DNA typing of the products of each culture step was conducted to assess purity. Serial exposure of MRSA COL to increasing concentrations of OXA resulted in increased resistance to ERY, which could be eliminated with RES. Evaluation of efflux pump activity by the EB method indicated increased efflux activity. Resistance to ERY was accompanied by resistance to kanamycin, amikacin, ofloxacin, norfloxacin, ciprofloxacin and rifampicin. This is the first time that a multidrug-resistant phenotype has been experimentally produced as a consequence of exposure of the organism to an antibiotic to which it is initially highly resistant.     

Analysis of mechanisms involved in reduced susceptibility to ciprofloxacin in Salmonella enterica serotypes Typhi and Paratyphi A isolates from travellers to Southeast Asia

Owing to multidrug resistance, quinolones and third-generation cephalosporins are currently used as key antibiotics to combat Salmonella organisms. Therapy failure due to reduced ciprofloxacin susceptibility has been reported in endemic areas, but also in imported disease. Different bacterial resistance mechanisms may result in reduced ciprofloxacin susceptibility. In this study, the presence and expression of different resistance mechanisms resulting in reduced minimum inhibitory concentrations (MICs) for ciprofloxacin were evaluated in 23 blood-culture-derived Salmonella enterica serotypes Typhi and Paratyphi A organisms from ill-returned travellers to Asia. The presence of mutations in the quinolone resistance-determining region (QRDR) of the gyrA gene as well as an activated efflux pump and plasmid-mediated quinolone resistance genes was determined. Resistance selection during therapy and the clonal relatedness of all isolates were established. Efflux pump inhibition did not appear to affect the MICs of ciprofloxacin and activity of the efflux pump appeared to be specific for nalidixic acid. Repeated exposure of the isolates to ciprofloxacin did not result in a significant increase in the MICs for ciprofloxacin. Repetitive sequence-based polymerase chain reaction (rep-PCR) profiles identified five different genotypes, but no correlation with resistance was observed. However, a significant relation was found with geographic region; reduced ciprofloxacin susceptibility was only found in travellers returning from India and Pakistan. All isolates with reduced ciprofloxacin susceptibility had a mutation at position 83 in the QRDR region of the gyrA gene. Plasmid-mediated quinolone resistance was not found. These findings confirm that the reduced ciprofloxacin MIC in S. Typhi and S. Paratyphi A is solely due to an amino acid substitution in the QRDR 'cluster' of the gyrA gene.     

Detection of decreased susceptibility to fluoroquinolones in Salmonella spp. by five different methods including real-time polymerase chain reaction (PCR)

Detection of Salmonella spp. isolates showing decreased susceptibility to fluoroquinolones has become important owing to the increasing prevalence of these strains and their association with treatment failure. Nalidixic acid agar dilution, nalidixic acid disk diffusion, MicroScan automated system and real-time polymerase chain reaction (PCR) (LightCycler) followed by melting temperature (Tm) analysis are compared with ciprofloxacin agar dilution as suitable methods to detect decreased susceptibility to fluoroquinolones in 100 Salmonella spp. isolates. Three minor discrepancies were found for nalidixic acid disk diffusion, one minor discrepancy was found for nalidixic acid agar dilution and Tm analysis, and one major discrepancy was found for MicroScan. Nalidixic acid disk diffusion was confirmed as a good screening method. Tm analysis is a rapid and accurate method for detecting decreased susceptibility to fluoroquinolones due to gyrA mutations in Salmonella spp.     

Quinolone pharmacokinetics

Fluoroquinolones have broad antibacterial spectra and are active against most Gram-negative and many Gram-positive species. They exhibit excellent oral bioavailability, extensive tissue penetration, low protein binding, and a long elimination half-life. This review compares and contrasts the pharmakonetics of some quinolone antibiotics - especially pefloxacin, ciprofloxacin, enoxacin, norfloxacin, ofloxacin, fleroxacin and lomefloxacin - in terms of their adsorption, distribution, metabolism, elimination, and interactions with other drugs and with food. In addition, the pharmacokinetics of these agents in the elderly and in patients with renal or hepatic impairment is discussed. The fluoroquinolones are established as a major class of antibiotics in the treatment of infections but pharmacokinetics factors should be considered when deciding on the most appropriate of these agents to use in individual patients.     

六种氟喹诺酮对肠球菌的体外抗菌活性及利血平的影响

目的：研究氟喹诺酮类抗菌药物对临床分离肠球菌的体外抗菌活性,以及多重耐药泵抑制剂利血平对抗菌活性的影响.方法：收集临床分离的101株肠球菌（66株粪肠球菌和35株屎肠球菌）,用琼脂稀释法测定应用利血平前后6种氟喹诺酮对菌株的最低抑菌浓度（MIC）.结果：诺氟沙星、环丙沙星、氧氟沙星、左氧氟沙星、加替沙星、莫西沙星对66株粪肠球菌的MIC90依次为256、64、64、16、16、8 mg/L,对35株屎肠球菌的MIC90依次为＞512、512、128、128、32、32 mg/L.应用利血平之后,上述6种药物对粪肠球菌抗菌活性提高（MIC下降2倍或2倍以上）的株数依次为66（100%）、54（81.8%）、4（6.1%）、4（6.1%）、32（48.5%）和3（4.5%）株,对屎肠球菌抗菌活性提高的株数依次为35（100%）、29（82.9%）、1（2.9%）、0（0%）、6（20.7%）和2（5.7%）株.结论：新氟喹诺酮加替沙星、莫西沙星增强了对肠球菌的抗菌活性,利血平能够提高全部或部分被检测肠球菌对诺氟沙星、环丙沙星和加替沙星的敏感性,但仅使少数被检测肠球菌对氧氟沙星、左氧氟沙星和莫西沙星的敏感性提高.     

The simultaneous separation and determination of five quinolone antibiotics using isocratic reversed-phase HPLC: application to stability studies on an ofloxacin tablet formulation

A rapid and reliable HPLC method was developed for the simultaneously separation and quantitation of five quinolones antibiotics; nalidixic acid, norfloxacin, ofloxacin, ciprofloxacin and lomefloxacin. All five tablet formulations of individual quinolone antibiotics were routinely assayed without interference. The calibration curves were linear (r2> or =0.999) over the concentration range of 1.20-4.8 mg/100 ml. Selectivity, precision, sensitivity and accuracy were established and the method is stability indicating with respect to ofloxacin. The limit of detection and quantitation for ofloxacin was 18 and 36 microg/100 ml, respectively. The separation was performed on a Phenomenex ODS C18 column using an isocratic, ion-pairing mobile phase consisting of 35% (v/v) aqueous acetonitrile together with tetrabutylammonium acetate, sodium dodecyl sulphate and citric acid (pH* 3.4). All analyses were conducted at ambient temperature and was monitored using a Diode Array UV/VIS detector set at wavelengths 235, 254, 275 and 300 nm.     

The bactericidal activity of levofloxacin against ampicillin-resistant and ampicillin-susceptible Haemophilus influenzae in comparison with ofloxacin, ciprofloxacin and sparfloxacin

The bactericidal activity of levofloxacin against four Haemophilus influenzae clinical isolates (two ampicillin-resistant and two susceptible) was compared with that of ofloxacin, ciprofloxacin and sparfloxacin at concentrations simulating the peak serum concentrations obtained with the recommended oral doses. Bactericidal activity was assessed using time-kill curves and minimum kill-time values. Both concentrations of levofloxacin rapidly killed all the study strains, with mean kill times of 4 h and no viable bacteria remaining after 18-h exposure. The bactericidal activities of levofloxacin, ofloxacin and sparfloxacin were similar. The minimum kill-times for both concentrations of ciprofloxacin were 28-35% longer than those of levofloxacin. These results support the use of levofloxacin for H. influenzae infections, including ampicillin-resistant strains.     

Antibiotic resistance in Salmonella enterica subsp. enterica strains isolated in Poland from 1998 to 1999.

A total of 326 Salmonella enterica subsp. enterica strains representing 29 serotypes, isolated from human stool specimens during 1998-1999 in sanitary-epidemiological units in Poland were tested for antibiotic susceptibility by a standard disk diffusion method. The antibiotics used were ampicillin, cefotaxime, chloramphenicol, tetracycline, streptomycin, gentamicin, kanamycin, nalidixic acid, ciprofloxacin, furazolidone, cotrimoxazole, sulphonamides and trimethoprim. In addition, 201 strains belonging to the five most commonly isolated serotypes (S. Enteritidis, S. Typhimurium, S. Hadar, S. Infantis and S. Virchow) also had minimal inhibitory concentrations (MICs) determined for amoxycillin/clavulanic acid. Selected strains were screened for production of extended spectrum beta-lactamases (ESBLs). There were 49.4% of Salmonella enterica subsp. enterica strains resistant to two or more antibiotics, with the highest prevalence of multiple resistant strains among serotypes Typhimurium, Hadar and Virchow. Resistance to ampicillin, streptomycin, tetracycline, nalidixic acid, furazolidone and sulphonamides occurred most frequently. Over 93% of S. Virchow strains were resistant to furazolidone. No strains resistant to ciprofloxacin by disk-diffusion method were detected but 31.3% of isolates of the 201 strains representing the five most common serotypes had reduced ciprofloxacin susceptibility (MICs ranging 0.125-0.5 mg/l). One strain (S. Mbandaka) was resistant to cefotaxime and produced ESBL.     

Occurrence and ecological risk assessment of fluoroquinolone antibiotics in hospital waste of Lahore,Pakistan

In the present study, wastewater and sludge samples of two major hospitals of Lahore, Pakistan were analyzed by developing an HPLC-UV method for the possible occurrence of five frequently used fluoroquinolone antibiotics i.e. ofloxacin, ciprofloxacin, sparfloxacin, moxifloxacin and gemifloxacin. The highest detected concentration was for moxifloxacin in both wastewater (224 μg/L) and sludge samples (219 μg/kg. The highest concentration of ofloxacin, ciprofloxacin, sparfloxacin and gemifloxacin were found to be 66, 18, 58 and 0.2 μg/L respectively. Risk quotient (RQ) was also calculated based on maximum measured concentrations and the RQ values were very high particularly for ofloxacin and ciprofloxacin. The maximum RQ values for ofloxacin against Vibrio fisheri, Pseudomonas putida, fish, Daphnia, Green algae and Pseudokirchneriella subcapitata were 3300, 66,000, 124, 46, 3300 and 6000, respectively. In case of ciprofloxacin, RQ values were found to be 1750 and 3500 against green algae and Microcystis aeruginosa, respectively.     

The comparative activity of twelve 4-quinolone antimicrobials against gram-positive and gram-negative anaerobes

The minimal inhibitory concentrations (MICs) of twelve 4-quinolone antimicrobials were determined for the Bacteroides fragilis group (50), Bacteroides melaninogenicus (20), Bacteroides bivius (10), Fusobacterium spp. (10), anaerobic Gram-positive cocci (50) and Clostridium spp. (20). MICs were determined using an agar dilution technique in Mueller-Hinton agar supplemented with 10% lysed horse blood. The inoculum used was approximately 10(4) colony-forming units, contained in 10 microliter of Mueller-Hinton broth, which was applied to the agar plates using a multipoint inoculator. Following inoculation, plates were incubated at 37 degrees C for 48 h in an anaerobic atmosphere. The MIC of each antimicrobial for each isolate examined was determined as the lowest concentration of the antimicrobial which completely inhibited growth of the inoculum. The minimum concentrations required to inhibit the growth of 50% (MIC50) and 90% (MIC90) of the organism examined were also determined. All of the more recently synthesised 4-quinolones showed increased activity against the anaerobic bacteria used in this study. Ciprofloxacin and ofloxacin were the most active compounds examined (Bacteroides fragilis group MIC90 ciprofloxacin 4 micrograms/ml; ofloxacin 4 microgram/ml; Bacteroides melaninogenicus MIC90 ciprofloxacin 2 micrograms/ml, ofloxacin 2 micrograms/ml; Bacteroides bivius MIC90 ciprofloxacin 16 micrograms/ml, ofloxacin 32 micrograms/ml; Fusobacterium spp. MIC90 ciprofloxacin 2 micrograms/ml, ofloxacin 4 micrograms/ml; Clostridium spp. MIC90 ciprofloxacin 1 microgram/ml, ofloxacin 1 microgram/ml and anaerobic Gram-positive cocci MIC90 ciprofloxacin 4 micrograms/ml, ofloxacin 4 micrograms/ml).