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In-vitro study of the activity of ciprofloxacin alone and in combination against strains of Pseudomonas aeruginosa with multiple antibiotic resistance.

Ciprofloxacin appears to have useful activity against Pseudomonas aeruginosa. We have studied its in-vitro activity against ten strains of Ps. aeruginosa with multiple antibiotic resistance. We have confirmed that ciprofloxacin is very active against Ps. aeruginosa with minimal inhibitory concentrations ranging from 0.07 to 0.7 mg/l. Killing curves show ciprofloxacin to be rapidly bactericidal with no regrowth after 24 h. Checkerboard studies with ciprofloxacin in combination with gentamicin, azlocillin and ceftazidime show no consistent interaction. These studies suggest that ciprofloxacin should prove a useful antibiotic in treating infections caused by multiresistant Ps. aeruginosa.

Anti-Bacterial Agents↗

Amikacin synergism with beta-lactam antibiotics against selected nosocomial pathogens.

The synergistic interaction between amikacin and several investigational antibiotics against seven different genera of nosocomial pathogens was assessed using the microtitre checkerboard technique. The greatest percentage of tests showing synergy was when amikacin was used in combination with apalcillin and azlocillin particularly against Serratia marcescens and Proteus spp. When amikacin was combined with several new semi-synthetic cephalosporins, synergy was present in a variable percentage of tests. No antagonism was found in any of the tested combinations.

Amikacin↗

Susceptibility of Pseudomonas aeruginosa strains isolated from hospitalized children.

Pseudomonas aeruginosa strains, 1285 in total, were isolated from hospitalized children in the period 1981-1984. Serotypes 011 and 06 and nontypable strains were the most frequently identified. 44.6% of serotype 011 strains and between 12.4 and 37.7% of other serotypes were resistant to gentamicin, or carbenicillin, or both agents. The aminoglycosides with the highest activity against P. aeruginosa isolates were amikacin and netilmicin. Among the seven beta-lactam antibiotics tested the most active were cefsulodin, piperacillin and azlocillin.

Aminoglycosides↗

Synergy of ciprofloxacin with fosfomycin in vitro against Pseudomonas isolates from patients with cystic fibrosis.

Pseudomonas aeruginosa remains the most common respiratory pathogen causing morbidity and mortality in cystic fibrosis (CF) patients. The in-vitro activity of ciprofloxacin and fosfomycin (calcium and tromethamine salts) in combination against P. aeruginosa isolates from CF patients, all of whom had received previous courses of ciprofloxacin, was evaluated by agar plate dilution chequerboard technique. The concentrations of drugs used were those that would be achieved in patients by oral and intravenous (fosfomycin only) routes. Synergy, taking into account fosfomycin concentrations achievable intravenously, was found against 60% of P. aeruginosa isolates. With concentrations achieved after an oral dose, 17% of isolates were synergistically inhibited. Time-kill experiments confirmed these findings. Ciprofloxacin MICs against resistant P. aeruginosa were reduced to achievable sputum and serum levels in the presence of fosfomycin. However, in progressive resistance studies fosfomycin failed to delay the development of resistance to ciprofloxacin. The combinations of ofloxacin/fosfomycin and ciprofloxacin/fosmidomycin were also tested, but showed minimal synergy. No antagonism was observed with any combination. Fosfomycin, ciprofloxacin and azlocillin in triple combination did not show synergy. The antimicrobial combinations tested against P. cepacia isolates from CF patients were indifferent. The combination of ciprofloxacin and fosfomycin may be clinically useful in selected P. aeruginosa pulmonary exacerbations in cystic fibrosis patients, particularly as an oral out-of-hospital treatment alternative or in cases where MICs for ciprofloxacin are elevated.

Ciprofloxacin↗

The antimicrobial susceptibility of Pseudomonas pseudomallei. Emergence of resistance in vitro and during treatment.

We have measured the in-vitro activity of 27 antimicrobials against 211 clinical and ten reference strains of Pseudomonas pseudomallei. Imipenem was the most active antibiotic tested, followed by piperacillin, doxycycline, amoxycillin/clavulanic acid, cefixime, cefetamet, azlocillin and ceftazidime, all of which had MICs of less than or equal to 2 mg/l for the majority of strains. The measured MICs were dependent on the media and inocula used, to an extent which varied with the antibiotic class under test; MICs of ureidopenicillins were particularly inoculum-dependent. The beta-lactams and ciprofloxacin were bactericidal, whereas the agents conventionally used to treat melioidosis (doxycycline, chloramphenicol, sulphamethoxazole and trimethoprim) had bacteriostatic activity only. Strains highly resistant to chloramphenicol (MIC greater than or equal to 256 mg/l) emerged during treatment in 7.1% of patients. These strains were fully virulent, and frequently showed cross-resistance to tetracyclines, sulphamethoxazole, trimethoprim and ciprofloxacin, with paradoxical increased susceptibility to beta-lactams and aminoglycosides. Similar resistance patterns were seen in mutants generated in vitro and two reference strains. One strain with isolated ceftazidime resistance, reversible by clavulanic acid, emerged during treatment. Several of the new beta-lactam antibiotics are of potential value in the therapy of P. pseudomallei infections. Patients should be carefully monitored for the emergence of antibiotic-resistant strains during treatment of melioidosis.

Anti-Bacterial Agents↗

Resistance mechanisms of multiresistant serotype 012 Pseudomonas aeruginosa isolated in Europe.

Serotype 012 Pseudomonas aeruginosa resistant to gentamicin (MIC greater than 4 mg/l) and carbenicillin (MIC greater than 128 mg/l) occur widely in Europe and are homogeneous in their phenotypic and genetic properties. It has been suggested that a single multiresistant strain of this serotype has become widespread. This study examined the resistance mechanisms present in multiresistant serotype 012 P. aeruginosa isolates from Austria, Belgium, France, Greece, Italy, Holland, West Germany and the UK. Disseminated isolates produced a PSE-1 type beta-lactamase, correlating with their resistance to the known substrates of this enzyme (carbenicillin, azlocillin and cefsulodin). These isolates also reacted with gene probes for the aminoglycoside modifying enzymes AAC(6')I and ANT(3'). The probe for AAC(6')I is known to cross-react with the gene for AAC(6')II. The fact that the organisms were resistant to netilmicin, gentamicin, sisomicin and tobramycin, but less so to amikacin, suggested that the latter enzyme was produced rather than AAC(6')I. PSE-1 beta-lactamase and the gene for AAC(6')I/AAC(6')II were absent from the International Antigenic Typing Scheme 012 reference strain, which was sensitive to beta-lactams and aminoglycosides, and also from a multiresistant serotype 012 strain isolated at a London burns unit in 1987. These organisms have been shown previously to be distinct from the disseminated multiresistant strain in their phenotypic properties. Two 012 isolates appeared to have additional resistance determinants to amikacin and isepamicin.

Aminoglycosides↗

Tolerance and safety of ciprofloxacin in paediatric patients.

The Cystic Fibrosis Clinic at the Royal Belfast Hospital for Sick Children has treated 31 children with ciprofloxacin, for serious pseudomonas infection in cystic fibrosis, and carefully monitored the safety and acceptability of the drug. Initially, eight very ill children were treated on a named-patient basis, with an encouraging clinical response and few adverse effects. Children aged 10-18 years were included in a study of four consecutive exacerbations of respiratory disease, comparing (i) oral ciprofloxacin in each episode with (ii) ciprofloxacin alternating with intravenous azlocillin and tobramycin. Other children with cystic fibrosis were subsequently treated with ciprofloxacin, as the need arose. In all the groups very few adverse reactions were found; in particular only one child developed arthralgia. A total of 202 children in the UK have been treated with ciprofloxacin on a named-patient basis, and their clinicians have reported 46 adverse events that may have been drug-related. Overall ciprofloxacin appears to be safe and effective in children but concern about the possible occurrence of arthropathy remains and long term follow-up of these children may be necessary.

Adolescent↗

Decreased susceptibility of penicillin-resistant pneumococci to twenty-four beta-lactam antibiotics.

The in-vitro activity of 24 beta-lactam antibiotics was compared using three groups of pneumococci and an agar dilution method comprising 100 penicillin-susceptible, 100 intermediately penicillin-resistant, and 100 highly penicillin-resistant pneumococcal strains. Our results show that intermediately penicillin-resistant and highly penicillin-resistant pneumococci had decreased sensitivity to other beta-lactam agents. According to their relative in-vitro activity, the antimicrobials were classified into three groups. The first group included drugs more active than penicillin (imipenem, meropenem, cefotaxime, ceftriaxone, and cefpirome), which could be useful for the treatment of infections due to penicillin-resistant strains. The second group showed slightly lesser activity than did penicillin, and included: ampicillin, cefdinir, cefuroxime, cefoperazone, azlocillin, mezlocillin, piperacillin, cephalothin, and cefamandole. The remaining antibiotics (oxacillin, cefixime, ceftizoxime, cefetamet, cefaclor, ceftazidime, cefoxitin, cefonicid, and latamoxef) showed poor activity against penicillin-resistant strains, precluding their use for empirical treatment in areas with a high prevalence of penicillin-resistant strains.

Anti-Bacterial Agents↗

Heat stability of the antimicrobial activity of sixty-two antibacterial agents.

Sixty-two antimicrobial agents, including several combinations, were examined for stability at 56 degrees C for 30 min and 121 degrees C for 15 min, respectively. A microtiter broth dilution MIC test and an agar disk diffusion test served to test each chemo-agent for residual antimicrobial activity. Eleven drugs were partially heat-labile (MICs raised four- to eight-fold after autoclaving) and 26 drugs were heat-labile (MICs raised > or = 16-fold following autoclaving); the remainder proved heat-stable (MICs raised < or = two-fold after autoclaving). Surprisingly, the beta-lactams, azlocillin, aztreonam, mezlocillin, and oxacillin, were remarkably heat-stable.

Anti-Bacterial Agents↗

National survey of susceptibility to antimicrobials amongst clinical isolates of Pseudomonas aeruginosa.

Between September and December 1993, each of 24 hospitals in the UK collected up to 100 consecutive clinical isolates of Pseudomonas aeruginosa and sent these to the London Hospital Medical College (LHMC). Of 2184 cultures received, 1991 contained viable P. aeruginosa. Minimum inhibitory concentrations (MICs) of antimicrobials were determined by agar dilution. The frequencies of resistance to low breakpoints were as follows: gentamicin, MIC > 2 mg/L, 11.7%; amikacin, MIC > 4 mg/L, 10.5%, carbenicillin, MIC > 128 mg/L, 11.7%; azlocillin, MIC> 16 mg/L, 10.9%; ceftazidime, MIC > 4 mg/L, 9.6%; ciprofloxacin, MIC > 1 mg/L, 8.1%; imipenem, MIC > 4 mg/L 2.5% and meropenem, MIC > 4 mg/L, 1.1%. Resistance to each antimicrobial except amikacin was commoner among the 134 isolates from patients in intensive care units (ICUs) than amongst the 1042 isolates from other in-patients (P < 0.01). Resistance to penicillins and ceftazidime, though not to other agents, was rarer among the 797 isolates from out-patients than amongst those from non-ICU in-patient (P < 0.01). Compared to a similar study in 1982, during which 1866 isolates had been examined, the frequency of resistance to the aminoglycosides increased (P < 0.05) as had those to the penicillins and ceftazidime (P < 0.01). Ciprofloxacin and the carbapenems were not tested in 1982. Cross-resistance patterns suggested that the increases in resistance to aminoglycosides and beta-lactams were largely a reflection of greater numbers of isolates with barrier or efflux mechanisms and were not due to an increase in isolates with antibiotic-degrading enzymes. The participating hospitals mostly employed Stokes' disc diffusion method and, when the results were compared to the MICs determined at the LHMC, fewer than 9% of the isolates reported as susceptible were found to be resistant. However, up to 72% of those reported by the hospitals as resistant were found to be susceptible.

Anti-Bacterial Agents↗

Absence of a post-antibiotic effect (PAE) of beta-lactams against Helicobacter pylori NCTC 11637.

A continuous culture system (chemostat) was used to study the post-antibiotic effect (PAE) of beta-lactams, against slowly-growing Helicobacter pylori NCTC 11637. H. pylori was grown at one quarter of its maximum specific growth rate (mu(max)) before exposure to ampicillin, amoxycillin, azlocillin, piperacillin or cefixime (8 x MIC). After 8 h, the antibiotics were inactivated. Viable counts were used to determine the rate of recovery of H. pylori. The recovery growth rate of H. pylori was similar to the maximum growth rate of H. pylori under antibiotic-free experimental conditions, so none of the beta-lactams studied showed a PAE against slowly-growing H. pylori.

Ampicillin↗

Analysis of antibiotic stability in a parenteral nutrition solution.

In children receiving multiple antibiotics and total parenteral nutrition (TPN), the amount of nutrition received can be less than optimal if the central venous line is used for administration of blood products, antibiotics and other medications. The purpose of this study was to evaluate the compatibility of commonly used antibiotics in our standard hyperalimentation solution to determine whether these drugs could be administered in a "piggyback" fashion with parenteral nutrition. If there were no incompatibility this could allow significantly more TPN to be delivered without need for extra fluid in patients receiving antibiotics several times daily. We found 13 antibiotics (amikacin, azlocillin, cefamandole, cephalothin, gentamicin, mezlocillin, moxalactam, nafcillin, oxacillin, penicillin, piperacillin, ticarcillin and tobramycin) to be stable for 6 hours and compatible with the TPN solution. They could be safely given in the presence of the hyperalimentation preparation (1.5% amino acid, 15% dextrose, vitamins, calcium (300 mg/liter) and standard electrolyte concentrations).

Anti-Bacterial Agents↗

The use of aztreonam in the cystic fibrosis patient.

Eradication of pulmonary infection by Pseudomonas aeruginosa in cystic fibrosis (CF) patients has long presented a significant challenge to the medical community. Many antimicrobial agents have proved incompletely effective against this persistent pathogen, and even the aminoglycosides, which represent the traditional therapy for such infections, have been associated with considerable toxicity and resistance. The monobactam antibacterial agent aztreonam is used both as single-agent therapy and in combination with other drugs. Several controlled, clinical trials have demonstrated the efficacy of aztreonam in the treatment of CF patients with pulmonary exacerbations caused by P. aeruginosa. The only side effect of aztreonam therapy commonly encountered in these studies was elevation of hepatic transaminase concentrations; this effect was of uncertain significance. It was concluded that aztreonam may offer clinical efficacy comparable to that provided by the combination of tobramycin plus azlocillin. Further, there does not seem to be any appreciable difference in the development of resistance to aztreonam compared with traditional therapies.

Aztreonam↗

Dissociation of surface properties and "intrinsic" resistance to beta lactams in Pseudomonas aeruginosa.

Carbenicillin resistance in strains of Pseudomonas aeruginosa isolated in Britain is mediated more frequently by "intrinsic factors" than by beta-lactamase production. Intrinsically carbenicillin-resistant isolates almost invariably were more resistant to azlocillin, cefoperazone, cefotaxime, ceftazidime, chloramphenicol, tetracycline and nalidixic acid than were carbenicillin-susceptible strains. This cross-resistance to different classes of antimicrobials suggested an impermeability-based mechanism of resistance, perhaps involving the outer membrane. The structure and composition of the outer membrane of the pseudomonas cell also influences the O-serotype specificity and the susceptibility to many bacteriophages. We therefore examined these properties for possible relationships to antibiotic resistance. Carbenicillin-resistant (122) and -sensitive (127) P. aeruginosa strains from 24 hospitals were compared. Serotype O:1, O:3, O:6, O:10 and O:11 strains predominated in both groups. Non-typable and polyagglutinating strains were infrequent in both groups. With one possible exception, none of 18 bacteriophages showed a significant preference for carbenicillin-resistant or -sensitive strains. Variation between strains was observed in the electrophoretic profile of LPS and this could be related in part to serotype, but not to antibiotic resistance. Our results contrast with those of earlier small-scale studies which have claimed relationships between surface properties and antibiogram in P. aeruginosa, and suggest that interpretation of the minor changes in LPS sometimes observed in association with the development of antibiotic resistance in vitro requires caution.

Anti-Bacterial Agents↗

Interactions of the 4-quinolones with other antibacterials.

The effect of sub-inhibitory concentrations of 16 antibacterials on the bactericidal activity of the 4-quinolones nalidixic acid, ciprofloxacin and ofloxacin against Escherichia coli KL16 in nutrient broth was investigated. Sub-inhibitory concentrations of rifampicin, clindamycin, chloramphenicol, erythromycin or tetracycline antagonised the bactericidal activity of the 4-quinolones. Conversely, all seven aminoglycosides tested enhanced the bactericidal activity of the 4-quinolones whereas the cell wall antagonists, azlocillin, mezlocillin, ceftazidime and vancomycin had no effect on the bactericidal activity of the 4-quinolones.

Anti-Bacterial Agents↗

Correlation between antibiotic sensitivity testing by conventional and conductivity measurements.

A high correlation in the susceptibilities of 44 strains isolated from clinical material to a variety of antimicrobial agents was found between the minimum inhibitory concentration and conductivity measurements. Some discrepancy was found for vancomycin and teicoplanin when tested against 21 strains of staphylococci. Similarly, discrepancies were seen with azlocillin, chloramphenicol and gentamicin when tested against strains of aerobic and facultatively anaerobic Gram-negative bacilli. In the majority of cases in which discrepancies were noted, this reflected differences in relative, and not absolute, susceptibility. With five strains of Pseudomonas, however, there was a poor correlation with three of four antibiotics studied. The advantage of this technique is that sensitivities are available within 4-6 h.

Anti-Bacterial Agents↗

Comparative in vitro susceptibility of Bacteroides and Fusobacterium isolated from footrot in sheep to 28 antimicrobial agents.

The agar dilution method was used to determine the bacteriostatic activity of 28 antimicrobial agents against 141 strains to the genus Bacteroides and 29 strains from the genus Fusobacterium. All organisms were isolated from clinical cases of ovine footrot. The strains were isolated from 125 Merino sheep, over a period of 2 years, from January 1987 to December 1988. The three ureidopenicillins studied (azlocillin, mezlocillin and piperacillin) proved to be the most effective antimicrobial agents. Chloramphenicol, metronidazole and tinidazole effectively inhibited the growth of Bacteroides spp., while phosphomycin was active against Fusobacterium spp.

Animals↗

Searching for new antimalarial therapeutics amongst known drugs.

The need to discover and develop new antimalarial therapeutics is overwhelming. The annual mortality attributed to malaria, currently approximately 2.5 million, is increasing due primarily to widespread resistance to currently used drugs. One strategy to identify new treatment alternatives for malaria is to examine libraries of diverse compounds for the possible identification of novel scaffolds. Beginning with libraries of drug or drug-like compounds is an ideal starting point because, in the case of approved drugs, substantial pharmacokinetic and toxicologic data should be available for each compound series. We have employed a high-throughput screen of the MicroSource Spectrum and Killer Collections, a library of known drugs, bioactive compounds, and natural products. Our screening assay identifies compounds that inhibit growth of Plasmodium falciparum cultured in human erythrocytes. We have identified 36 novel inhibitors of P. falciparum, of which 19 are therapeutics, and five of these drugs exhibit effective 50% inhibitory concentrations within similar ranges to therapeutic serum concentrations for their recently indicated uses: propafenone, thioridazine, chlorprothixene, perhexiline, and azlocillin. The findings we report here indicate that this is an effective strategy to identify novel scaffolds and therefore aid in antimalarial drug discovery efforts.

Animals↗