In-vitro susceptibility of Pseudomonas pseudomallei to DNA gyrase inhibitors.
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Biomedical subjects
Publications and source records attributed to D Felmingham.
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In a prospective study serum C-reactive protein concentrations were measured in nine patients with "active" pulmonary sarcoidosis (as assessed by bronchoalveolar lavage lymphocyte counts, gallium-67 lung scanning, and serial pulmonary function testing), and in five patients with "inactive" disease. Active pulmonary sarcoidosis was associated either with no rise or with only a modest rise in serum C-reactive protein concentrations. In contrast, serum C-reactive protein concentrations in 12 patients with sputum positive pulmonary tuberculosis were considerably raised. Serum C-reactive protein may thus provide a valuable test in the differentiation of sarcoidosis from conditions which it may mimic and which are known to induce an acute phase response.
An accurate and sensitive bioassay for determining concentrations of teicoplanin in serum has been developed using modifications of procedures described for assaying vancomycin. A linear relationship (r = 0.9983) was obtained between the diameter of the zone of inhibition and log10 teicoplanin concentration over the range 0.25-32 micrograms/ml. The medium used was Antibiotic Medium No. 1 (Oxoid, UK) adjusted to pH 5.5-5.7 by the addition of hydrochloric acid and containing sodium chloride to a final concentration of 3%. The indicator organism used was a Bacillus subtilis ATCC 6633 (NCTC 10400). Teicoplanin was assayed: (i) in the presence of beta-lactams and cephalosporins, including ceftazidime, by prior treatment of serum with broad-spectrum beta-lactamase mixtures (Genzyme Laboratories, UK); (ii) in the presence of aminoglycosides by prior treatment of serum with cellu-ion phosphate (100 mg/ml) followed by centrifugation; (iii) in the presence of sulphamethoxazole and/or trimethoprim by the addition of p-aminobenzoic acid and/or thymidine to the assay medium. Teicoplanin was assayed in the presence of either rifampicin or erythromycin by using a rifampicin-resistant, erythromycin-resistant clinical isolate of Staphylococcus aureus as indicator organism. The lower limit of sensitivity of this assay was 1 microgram/ml. The presence of undeclared, broad-spectrum antimicrobials was detected by screening serum samples for antimicrobial activity using an assay plate seeded with Escherichia coli NCTC 10418.
The in vitro activity of vancomycin and teicoplanin, a new glycopeptide antimicrobial, was determined against a total of 286 anaerobic bacteria including Bacteroides fragilis group (100), B. melaninogenicus (21), B. bivius (16), Fusobacterium spp. (15), Peptococcus spp. (20), Peptostreptococcus spp. (21), Clostridium perfringens (23), C. difficile (41) and Propionibacterium acnes (29). Minimum inhibitory concentrations (MIC) were determined using an antimicrobial incorporation technique in Wilkins-Chalgren agar approximately 10(4) colony forming units (cfu) contained in 10 microliters Wilkins-Chalgren broth, which was applied to the surface of the agar plates using a multipoint inoculator. Following inoculation, plates were incubated for 48 h at 37 degrees C in an anaerobic atmosphere. Both vancomycin and teicoplanin were highly active against all the Gram-positive anaerobic bacteria examined, 90% of all isolates being inhibited by 0.5 micrograms/ml of either antimicrobial. Isolates of B. fragilis group and Fusobacterium spp. were resistant to vancomycin (MIC90 64 micrograms/ml) and teicoplanin (MIC90 128 micrograms/ml). Unexpectedly, isolates of B. melaninogenicus and B. bivius which were resistant to vancomycin (MIC90 64 and 128 micrograms/ml respectively) were sensitive to teicoplanin (MIC90 2 and 2 micrograms/ml respectively).
The in vitro activity of erythromycin and RU-28965 (a novel macrolide antimicrobial with improved pharmacokinetics) was determined against a variety of anaerobic bacteria in anaerobic atmospheres with and without added carbon dioxide. Minimum inhibitory concentrations (MIC) were determined using an antimicrobial incorporation technique in Wilkins-Chalgren agar (Oxoid, UK) containing saponinlysed horse blood to a final concentration of 10%. The inoculum used was approximately 10(4) colony forming units (cfu) contained in 10 microliters Wilkins-Chalgren broth, which was applied to the surface of the agar plates using a multipoint inoculator. Following inoculation, plates were incubated for 48 h at 37 degrees C in an anaerobic atmosphere containing 10% carbon dioxide or in hydrogen alone. The MIC of each antimicrobial for each organism 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 bacteria examined were also determined. The MICs of erythromycin and RU-28965 for isolates of the Bacteroides fragilis group, B. bivius and Fusobacterium spp. were generally 10-100 times greater when determined in the presence of carbon dioxide than when determined in hydrogen alone. The MICs of erythromycin and RU-28965 for B. melaninogenicus, Peptococcus spp., Peptostreptococcus spp., Clostridium perfringens, Cl. difficile and Propionibacterium acnes were less affected by the presence of carbon dioxide.
The in vitro activity of aztreonam, cefuroxime and ceftazidime was determined against 2,372 Gram-negative rods (including Pseudomonas spp.) isolated from hospital patients with urinary tract infections during 1985. Minimum inhibitory concentrations (MICs) were determined using an agar incorporation technique in Mueller-Hinton agar. The inoculum used was approximately 10(5) colony forming units (cfu) contained in 10 microliter Mueller-Hinton broth, which was applied to the surface of the agar plates using a multipoint inoculator. Following inoculation plates were incubated aerobically at 37 degrees C for 18 h. The MIC of each antimicrobial for each organism examined was determined as the lowest concentration of the antimicrobial which completely inhibited growth of the inoculum. The minimum concentration required to inhibit the growth of 90% (MIC90) of the bacterial isolates in each genus or species examined was also determined. In general the antibacterial spectrum of aztreonam was comparable to that of ceftazidime and superior to that of cefuroxime. Against Escherichia coli, which accounted for 72% of the isolates examined, aztreonam (MIC90 less than or equal to 0.25 microgram/ml) was slightly more active than ceftazidime (MIC90 0.5 microgram/ml) and considerably more active than cefuroxime (MIC90 8 micrograms/ml). Aztreonam was active against Pseudomonas spp. (MIC90 16 micrograms/ml), although somewhat less so than ceftazidime (MIC90 4 micrograms/ml). Cefuroxime showed low activity against this genus (MIC90 greater than 128 micrograms/ml).
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The Pauldrach Endocleaner was evaluated for the disinfection of gastrointestinal fibrescopes, and found to be a convenient, relatively inexpensive machine for use in endoscopy clinics. An experimental procedure was designed, for use with the endocleaner, which allowed application of an inoculum of Pseudomonas aeruginosa to a Fujinon OX52 endoscope and its subsequent quantitation before and after disinfection procedures. Using aldehyde based disinfectants ('Cidex', 'Kohrsolin' and 'Gigasept') it was possible to achieve a 99.999% reduction in the recovery of viable Ps. aeruginosa with a minimum exposure time of 20 min 'Dettox ABC' and buffered hypochlorite (100 ppm) were less effective achieving an approximately 99% reduction with a minimum exposure time of 30 min. Following apparently adequate disinfection and irrespective of the disinfectant used, the endoscope was found to be heavily contaminated with Ps. aeruginosa when examined after overnight storage at room temperature thus emphasising the need for meticulous disinfection of instruments prior to use in a day's list.
We have compared the in-vitro activity of 12 quinolone antibiotics against 50 recent isolates of Haemophilus ducreyi from The Gambia. While these isolates were relatively resistant to the parent compounds, the newly synthesized quinolones exerted significant activity against H. ducreyi. Ciprofloxacin and ofloxacin were the most active compounds (MIC90 0.03 mg/l).
The minimal inhibitory concentrations (MICs) of twelve 4-quinolone antimicrobials were determined for Salmonella typhi (25), Salmonella spp. (50), Shigella spp. (50), Campylobacter jejuni (100), Vibrio cholerae (10), Vibrio parahaemolyticus (10), Yersinia enterocolitica (25), Aeromonas hydrophila (25) and Plesiomonas shigelloides (10). MICs were determined using an agar dilution technique in Mueller-Hinton agar (Oxoid, England) supplemented with 10% lysed horse blood. Antibiotic containing plates were inoculated with approximately 10(4) colony forming units of each organism, contained in 10 microliters of Mueller-Hinton broth (Oxoid, England), using a multipoint inoculator. Following inoculation plates were incubated aerobically for 18 hours at 37 degrees C, except for plates inoculated with Campylobacter jejuni which were incubated microaerophilically for 48 hours at 37 degrees C. The MICs of each antimicrobial for each isolate examined, together with the minimum concentrations of each antimicrobial required to inhibit 50% (MIC50) and 90% (MIC90) of the isolates examined, were also determined. The more recently synthesized 4-quinolones showed very good activity against all of the enteric pathogens examined with ciprofloxacin being the most active (MIC90: Salmonella typhi 0.015 microgram/ml, Salmonella spp. 0.015 microgram/ml, Shigella spp. 0.015 microgram/ml, Campylobacter jejuni 0.12 microgram/ml, Vibrio cholerae 0.008 microgram/ml, Vibrio parahaemolyticus 0.06 microgram/ml, Yersinia enterocolitica 0.015 microgram/ml, Aeromonas hydrophila 0.015 microgram/ml and Plesiomonas shigelloides 0.015 microgram/ml. Where considered clinically appropriate these compounds may have a useful role in the treatment and prevention of diarrhoeal disease caused by these enteric pathogens.
The minimal inhibitory concentrations (MICs) of twelve 4-quinolone antimicrobials were determined for 100 isolates of Haemophilus influenzae (including 30 beta-lactamase producing strains) and 100 isolates of Streptococcus pneumoniae. 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 microliters 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 18 h in an atmosphere enriched to 10% carbon dioxide. 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 organisms examined were also determined. The more recently synthesised 4-quinolones showed considerably greater activity than nalidixic acid and pipemidic acid against clinical isolates of Haemophilus influenzae and Streptococcus pneumoniae. There was no apparent difference between the MICs observed for beta-lactamase producing and non-beta-lactamase producing strains of Haemophilus influenzae. Ciprofloxacin was the most active 4-quinolone examined (MIC90 for Haemophilus influenzae 0.008 microgram/ml; Streptococcus pneumoniae 2 micrograms/ml). Clinical studies on a possible role for some of the more recently synthesised 4-quinolones in the management of patients with respiratory infection are indicated.
The minimal inhibitory concentrations (MICs) of nalidixic acid, pipemidic acid, cinoxacin, oxolinic acid, flumequine, pefloxacin, acrosoxacin, amifloxacin, norfloxacin, enoxacin, ofloxacin and ciprofloxacin were determined for a range of clinical isolates. 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 microliters Mueller-Hinton broth, which was applied to the agar plates using a multipoint inoculator. Following inoculation, plates were incubated in conditions appropriate for the organisms under investigation. 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 organisms examined were also determined. All of the more recently synthesised 4-quinolones showed considerably greater activity than the parent compounds, nalidixic acid, pipemidic acid and cinoxacin, against the range of organisms used in this study. Ciprofloxacin and ofloxacin were the two most active of the 4-quinolones examined.
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).
The minimal inhibitory concentrations (MICs) of twelve 4-quinolone antimicrobials and sulphadiazine were determined for 160 clinical isolates of Neisseria meningitidis. The bacteria were recovered from nasopharyngeal carriers and cases of meningitis examined in The Gambia, West Africa, during the 1982-83 dry season. 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 of each organism, contained in 10 microliters of Mueller-Hinton broth, which was applied to the agar plates using a multipoint inoculator. Following inoculation, plates were incubated for 18 h at 37 degrees C in an atmosphere enriched to 5% carbon dioxide. 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 of each antimicrobial required to inhibit 50% (MIC50) and 90% (MIC90) of the isolates examined were also determined. The more recently synthesised 4-quinolones were very active against the isolates of Neisseria meningitidis, ciprofloxacin being marginally the most active (MIC90 0.008 micrograms/ml). The activity of the 4-quinolone antimicrobials was unaffected by the MICs of sulphadiazine required by the organisms, which ranged from 0.5- greater than 64 micrograms/ml.
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We report 11 patients with leishmaniasis from different endemic areas, treated in the UK with intravenous aminosidine alone or in combination with other drugs. Clinical and parasitological cures were achieved in all 7 patients from the Mediterranean zone who had visceral disease, with one relapse. Two of 4 patients with cutaneous or mucosal disease were cured; the other 2, from Iraq and Iran, did not respond. Toxic effects were high-tone deafness in 2 patients, one of whom had pre-existing renal impairment, and transient, mild elevation of serum creatinine in 3. Aminosidine is an effective, tolerable and relatively non-toxic alternative to existing antileishmanial drugs for the treatment of visceral leishmaniasis. Further studies will be needed to assess its place in cutaneous and mucosal disease.