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Two groups of Mycobacterium avium complex strains determined according to the susceptibility to rifampicin and ansamycin.

Mycobacterium avium complex strains previously not exposed to any antituberculosis agents could be divided into two groups according to their susceptibility to rifampicin and ansamycin; one group susceptible to 80 micrograms/ml rifampicin and to 1.25 micrograms/ml ansamycin, and another resistant to these concentrations. In each group, the ratio of the minimal inhibitory concentration of ansamycin against that of rifampicin was greatly different depending on the strain. This naturally occurring resistance to rifampicin and ansamycin was frequently correlated to naturally occurring resistance to ethambutol, kanamycin, enviomycin, kitasamycin, and minocycline, but not correlated to that to isoniazid and sulfadimethoxine. Ansamycin was more active than rifampicin against M. bovis, M. kansasii, M. marinum, M. xenopi, and M. haemophilum.

Anti-Bacterial Agents↗

[Effect of macrolide antibiotics on human serum-bactericidal sensitivity of Pseudomonas aeruginosa S-6].

It is well known that long-term administration of erythromycin (EM) at a small dose is effective for persistent infections with Pseudomonas aeruginosa in diffuse panbronchiolitis or chronic bronchitis patients. Since EM is less active against P. aeruginosa in vitro, we have been interested in the mechanisms of clinical efficacy of EM in these patients. This study examines the effect of macrolide antibiotics on human serum-bactericidal sensitivity of P. aeruginosa S-6, clinically isolated from the patient with respiratory tract infection. A significant increase in serum-bactericidal sensitivity of P. aeruginosa S-6 was observed on agar containing EM of 10 micrograms/ml after incubation for 36-60 hours (p less than 0.05). The enhancement of serum sensitivity of P. aeruginosa S-6 was apparently observed even at a concentration of EM 1.5 micrograms/ml after the 48 hours incubation (p less than 0.01). Of other macrolide antibiotics used, clarithromycin (CAM) also increased the serum-bactericidal sensitivity of P. aeruginosa S-6 as well as EM, however no change in the sensitivity was found with kitasamycin, josamycin, rokitamycin and oleandomycin. The results suggest that the change of serum-bactericidal sensitivity of P. aeruginosa induced by EM or CAM may, in part, contribute to the clinical efficacy of these antibiotics against persistent pulmonary P. aeruginosa infections.

Anti-Bacterial Agents↗

[Differences in susceptibility of Helicobacter pylori to macrolide and other antibiotics in tests using blood agar and albumin agar].

Minimum inhibitory concentrations (MICs) of penicillin G, cefazolin, tetracycline, chloramphenicol, four aminoglycosides, and seven macrolides against Helicobacter pylori strains were determined on Brucella agar supplemented with bovine serum albumin fraction V (albumin agar) and compared with those on blood agar. MICs of the macrolides against H. pylori determined on albumin agar were markedly lower than those found with blood agar. The susceptibilities of 26 H. pylori strains to the macrolides were estimated from the MIC50 values determined on blood agar and albumin agar, which were 0.39 and < or = 0.05 micrograms/ml for erythromycin, midecamycin and rokitamycin, 3.13 and 0.10 micrograms/ml for clindamycin, 1.56 and < or = 0.05 micrograms/ml for josamycin, 1.56 and 0.10 micrograms/ml for kitasamycin, and 0.10 and < or = 0.05 micrograms/ml for roxithromycin, respectively. However, the MICs of other antibiotics against H. pylori and of all antibiotics against two reference Campylobacter strains differed minimally between values obtained on blood agar and albumin agar.

Agar↗

Effects of rokitamycin and other macrolide antibiotics on Mycoplasma pneumoniae in L cells.

Fifty strains of Mycoplasma pneumoniae in L cells were tested for susceptibility to macrolide antibiotics. Rokitamycin, a new macrolide antibiotic, was most active against these organisms, with an MIC for 90% of strains of 0.007 microgram/ml. The MICs of erythromycin, josamycin, and kitasamycin for 90% of strains were 0.03, 0.03, and greater than or equal to 0.06 microgram/ml, respectively. Based on these results, rokitamycin is a promising antibiotic for the treatment of mycoplasmal infections, and further clinical investigations are needed.

Anti-Bacterial Agents↗

Comparison of methods for in vitro testing of susceptibility of porcine Mycoplasma species to antimicrobial agents.

The MICs of 18 antimicrobial agents used against strains of three porcine Mycoplasma species were determined by a serial broth dilution method. Twenty field strains of M. hyorhinis, ten field strains of M. hyopneumoniae, six field strains of M. flocculare, and the type strains of these species were tested. Twelve field strains and the type strain of M. hyorhinis were also tested by an agar dilution method. Tests were read at various time points. When the broth dilution method was used, the final MIC had to be read 2 days after color changes had stopped. MICs of tetracycline, oxytetracycline, doxycycline, and minocycline were low for the three Mycoplasma species tested. MICs of chlortetracycline were 8 to 16 times higher than MICs of the other tetracyclines. Spiramycin, tylosin, kitasamycin, spectinomycin, tiamulin, lincomycin, and clindamycin were effective against all strains of M. hyorhinis and M. hyopneumoniae. The quinolones were highly effective against M. hyopneumoniae but less effective against M. hyorhinis. The susceptibility patterns for M. hyopneumoniae and M. flocculare were similar.

Animals↗

Effects of sub-MICs of erythromycin and other macrolide antibiotics on serum sensitivity of Pseudomonas aeruginosa.

We examined the effects of sub-MICs of erythromycin (EM) and other macrolide antibiotics on the serum sensitivity of Pseudomonas aeruginosa. P. aeruginosa S-6 grown for 36 and 48 h on agar with 10 micrograms of EM per ml (1/10th the MIC) showed significantly increased sensitivity to human serum bactericidal activity compared with those of bacteria grown on agar without EM (P < 0.05). No changes in serum sensitivity were observed in bacteria grown for less than 24 h. This increased sensitivity was apparent even at a concentration of 1.5 micrograms of EM per ml (1/67th the MIC) in bacteria grown for 48 h (P < 0.01). Among the other macrolide antibiotics tested, clarithromycin also enhanced sensitivity to serum, but there were no changes in the sensitivities of bacteria grown on agar with kitasamycin, josamycin, rokitamycin, or oleandomycin even at a concentration of 12 micrograms/ml (1/16th, 1/16th, 1/8th, and 1/33rd the MICs, respectively). P. aeruginosa S-6 grown on agar with subinhibitory concentrations of EM showed decreased cell surface hydrophobicity in a dose-dependent manner, whereas oleandomycin and rokitamycin, even at a concentration of 12 micrograms/ml, induced a slight decrease in hydrophobicity which was approximately equivalent to that of 1.5 micrograms of EM per ml. Among six other strains of the nonmucoid phenotype, three strains became more sensitive to serum by exposure to 10 micrograms of EM per ml for 48 h. In contrast, no evident correlation between EM treatment and a change in serum sensitivity was observed in six strains of the mucoid phenotype, as judged by the results of experiments with both 2 and 0.4% serum. These results show that EM at subinhibitory concentrations enhances the serum sensitivity of some P. aeruginosa strains. Since induced serum sensitivity was accompanied by a decrease in bacterial cell surface hydrophobicity, EM may render P. aeruginosa more serum sensitive by changing the cell surface structure(s) of this organism.

Anti-Bacterial Agents↗

Induction of ermSV by 16-membered-ring macrolide antibiotics.

The erm family of 23S rRNA adenine-N6-methyltransferases confers resistance to all macrolide-lincosamide-streptograminB (MLS) antibiotics, but not all MLS antibiotics induce synthesis of Erm methyltransferase with equal efficiency in a given organism. The induction efficiency of a test panel of MLS antibiotics was studied by using two translational attenuator-lac reporter gene fusion constructs, one based on ermSV from Streptomyces viridochromogenes NRRL 2860 and the other based on ermC from Staphylococcus aureus RN2442. Four types of responses which were correlated with the macrolide ring size were seen, as follows: group 1, both ermSV and ermC were induced by the 14-membered-ring macrolides erythromycin, lankamycin, and matromycin, as well as by the lincosamide celesticetin; group 2, neither ermSV nor ermC was induced by the 12-membered-ring macrolide methymycin or by the lincosamide lincomycin or the streptogramin type B antibiotic ostreogrycin B; group 3, ermSV was selectively induced over ermC by the 16-membered-ring macrolides carbomycin, chalcomycin, cirramycin, kitasamycin, maridomycin, and tylosin; and group 4, ermC was selectively induced over ermSV by the 14-membered-ring macrolide megalomicin. These data suggest that the leader peptide determines the specificity of induction by different classes of MLS antibiotics and that for a given attenuator, a major factor which determines whether a given macrolide induces resistance is its size.

Anti-Bacterial Agents↗

Correlations among naturally occurring resistances to antituberculosis drugs in Mycobacterium avium complex strains.

In Mycobacterium avium complex strains, which were not exposed previously to any antituberculosis drugs, resistances to rifampin, minocycline, and kitasamycin, and resistances to streptomycin and kanamycin appeared frequently in the same strains. These data indicate that patterns of drug resistance in these organisms are not random but occur through linked mechanisms.

Animals↗

In vitro antibiotic susceptibility of field isolates of Mycoplasma synoviae in Argentina.

Minimum inhibitory concentrations (MICs) were determined in vitro for 7 antibiotics (aivlosin, enrofloxacine, tylosin, tiamulin, kitasamycin, chlortetracycline, and oxytetracycline) against eight recent local Argentinean isolates and two standard strains of Mycoplasma synoviae. Aivlosin (3-acetyl-4"-isovaleryl tylosin tartrate), tylosin, and tiamulin showed the lowest MICs with MIC90s of 0.006, 0.012, and 0.05 microg/ml, respectively. Except one strain that showed resistant values to chlortetracycline (> or = 12.5 microg/ml), all the analyzed strains were susceptible in different degrees to all the antibiotics tested. In this study, the improved activity of the tylosin-derived drug, aivlosin, was confirmed because it showed, in most strains, MIC values half those for tylosin.

Anti-Bacterial Agents↗

[Comparison of in vitro susceptibilities of Rickettsia prowazekii, R. rickettsii, R. sibirica and R. tsutsugamushi to antimicrobial agents].

The in vitro activities of 16 antimicrobial agents against Rickettsia prowazekii (Breinl strain), R. rickettsii (Bitterroot strain), R. sibirica (ATCC No. VR151) and R. tsutsugamushi (Gilliam, Karp, Kato, Shimokoshi, Kawasaki and Kuroki strains) were determined by the cell culture method. Tetracycline, demethylchlortetracycline, doxycycline, minocycline, chloramphenicol, kitasamycin and rifampicin were generally effective (MIC, 0.005-0.78 micrograms/ml) to all strains tested. Quinolones such as norfloxacin, ciprofloxacin and ofloxacin were moderately active, but they were less active against R. tsutsugamushi than other rickettsial species. Penicillins and cephems showed low activity against most of the strains tested, but high concentrations of benzylpenicillin (MIC, 25-50 micrograms/ml) inhibited R. prowazekii, R. rickettsii and R. sibirica. These findings may be applicable for differentiation of species of genus Rickettsia.

Anti-Bacterial Agents↗

Transduction of drug resistance to tetracycline, chloramphenicol, macrolides, lincomycin and clindamycin with phages induced from Streptococcus pyogenes.

Strains of Streptococcus pyogenes isolated from pediatric patients with acute infections which were resistant to one or more of the antibiotics, tetracycline (TC), chloramphenicol (CP), macrolide antibiotics (erythromycin, kitasamycin, oleandomycin, josamycin), lincomycin (LCM) and clindamycin (CLM), were used for transduction of drug resistance. These drug-resistant strains were treated with mitomycin C to induce phages and transduction of drug resistance was attempted by means of phages so induced. It was found that transduction of resistance to the above antibiotics was possible. The transductants obtained on TC-containing selective agar plate were resistant to TC alone while those produced on CP- or erythromycin (EM)-containing selective agar plate were resistant to CP, macrolide antibiotics (Mac), LCM and CLM. From this finding, it was inferred that transduction of resistance to TC, CP, Mac, LCM and CLM via phages occurred in two different patterns, i.e., transfer of resistance to TC alone and that of resistance to CP, Mac, LCM and CLM. All of the transductants obtained were found to belong to group A. In T-typing, they were of the same T-12 type as the donor and recipient strains in a majority of cases though some were not typable.

Anti-Bacterial Agents↗

[Epidemiological studies on drug-resistance patterns, coagulase types, and MRSA-phage types of MRSA isolates during 1990-1994].

We examined drug-resistance patterns, coagulase types, and MRSA-phage types of 125 MRSA strains isolated from clinical specimens during the period of January 1990 and December 1994. No vancomycin-resistant strain was isolated. Twenty one antibiotics were divided into three classes, low-intermediate- and high-isolation-frequency class, based on isolation frequencies of resistant strains. Minocycline, chloramphenicol, streptomycin, and imipenem were found to be included in low-isolation-frequency class (16.8-40%). In intermediate-isolation-frequency class (45.6-62.9%), cefmetazole, amikacin, gentamicin, and tetracycline were included. Oxacillin, ampicillin, piperacillin, ceftizoxime, cefoperazone, cefazolin, erythromycin, oleandomycin, kitasamycin, clindamycin, kanamycin, tobramycin, and ofloxacin belonged to high-isolation-frequency class (97.6-100%). MIC90s of vancomycin and minocycline (1.56 and 25 micrograms/ml) were lower than that of other 13 drugs. Comparing medical ward with dental ward, imipenem-, gentamicin-, and minocycline-resistant strains at medical ward, chloramphenicol- and streptomycin-resistant strains at dental ward were isolated dominantly on each ward, MRSA isolates were classified to 39 types by drug-resistance patterns. The isolation frequencies of coagulase type II and type IV strains were 65.6% and 29.6%, respectively. At dental ward, the isolation frequency of coagulase type IV strains was higher than that of coagulase type II strains during 1990-1992. However, coagulase type II strains were isolated considerably more than type IV strains during 1993-1994. By MRSA-phage typing, MRSA isolates were grouped into 18 MRSA-phage types. One hundred and twenty five MRSA isolates were divided into 56 types by using drug-resistance patterns, coagulase typing, and MRSA-phage typing. It was considered that such classification in combination of three methods is useful to make decision of epidemic by the same MRSA strain.

Anti-Bacterial Agents↗

[Antibacterial activity of rokitamycin against fresh clinical isolates].

We obtained bacterial strains which were clinically isolated and identified from outpatients with various infections in medical institutions throughout Japan. Possible antibacterial activities of rokitamycin (RKM) were examined against these isolates. Minimum inhibitory concentrations (MICs) were determined through a comparative study with reference drugs. The results of the study are summarized as follows. 1. Resistance patterns of 400 isolates which were highly resistant to macrolides (MLs) with MIC values > 100 micrograms/ml were classified into 55 patterns. Staphylococcus spp. showed cross resistance to 14-membered ring MLs with 100% cross resistance observed between erythromycin (EM) and clarithromycin (CAM), and 85.2% between EM and oleandomycin (OL). Fewer isolates showed strong resistance to 16-membered ring MLs than to 14-membered ring MLs. Cross resistances observed among the Staphylococcus isolates were 100% between acetylmidecamycin (MDM-AC) and kitasamycin (leucomycin (LM)), 93.9% between MDM-AC and josamycin (JM), and 53.3% between MDM-AC and RKM. Streptococcus spp. and Peptococcus spp. showed very similar resistance patterns to both 14- and 16-membered ring MLs, but resistance patterns to RKM were quite different. Most of anaerobic streptococci and Bacteroides fragilis group had similar resistance patterns to 14- and 16-membered ring MLs, but in some cases a pattern similar to that of Staphylococcus spp. was observed. 2. When ML-resistant bacteria isolated during 1975 to 1980 were compared to those isolated in 1986 and 1989, it was observed that resistance of Staphylococcus aureus remained almost unchanged, that of Streptococcus pyogenes was lower in the later years than during 1975 to 1980, but that of Streptococcus pneumoniae increased. 3. Most of ML-resistances of the resistant isolates were inducible, but extents of induction varied depending on drugs tested. Strong inductions were observed when 14-membered ring MLs were used, but inductions were minimal with 16-membered ring MLs. RKM appeared to induce resistance to the least extent. From these results, it appears that the RKM is quite useful clinically even in the 1990s.

Bacterial Infections↗

[Study on susceptibilities of freshly isolated strains to macrolide antibiotics].

Macrolide (ML) susceptibilities of clinically isolated bacteria obtained freshly during 1989 were determined and year to year differences in ML-resistances of the clinical isolates in our laboratory were examined with epidemiological consideration in mind. 1. The distribution of minimum inhibitory concentrations (MICs) of MLs against isolates of Staphylococcus aureus showed 2 peaks, one peak representing S. aureus isolates susceptible to MLs and the other peak representing those resistant to MLs. Proportions of resistant isolates to various MLs were 36% to erythromycin, 24% each to kitasamycin, josamycin, midecamycin and midecamycin acetate. MICs of rokitamycin (RKM), however, distributed themselves into 3 peaks of less than or equal to 0.39 micrograms/ml (76%), 1.56-3.13 micrograms/ml (12%) and greater than or equal to 100 micrograms/ml (12%). Staphylococcus spp. including Staphylococcus epidermidis showed different resistance characteristics to 14-membered ring MLs and to 16-membered ring MLs. Further, the proportion of resistant organisms to RKM was the smallest among proportions of resistant organisms to various 16-membered ring MLs tested. These differences in proportions of resistant organisms to different MLs may have resulted from differences in resistance induction activities of different MLs. The above described ratios of resistant organisms to MLs among S. aureus did not vary since 1975 among clinical isolates tested in our laboratory. 2. Four per cent (4%) of Streptococcus pyogenes isolates were MLs resistant, and 18% of Streptococcus pneumoniae isolates were resistant to MLs. Among the isolates tested in our laboratory since 1985, resistant isolates of S. pyogenes are clearly decreasing and those of S. pneumoniae seem to be increasing.(ABSTRACT TRUNCATED AT 250 WORDS)

Anti-Bacterial Agents↗

[Cross-resistance relationships of antituberculosis drugs in Mycobacterium avium complex].

At present, infection caused by Mycobacterium avium complex is usually treated by chemotherapy of antituberculosis drugs. However, cross-resistance relationships of antituberculosis drugs in the M. avium complex have not yet been studied. In the present study, we studied on this subject using three strains which were isolated from sputum specimens of patients who were not treated by any antituberculosis drugs: strain 13008 (serotype 20), strain 13016 (serotype 4) and strain 13034 (serotype 18). The methods used for isolating mutants resistant to rifampicin, isoniazid, ethambutol, streptomycin, kanamycin and/or enviomycin were described previously (Tsukamura, M: Kekkaku 62: 445-458, 1987). Mutants resistant to ethionamide were isolated from strains 13008 and 13016 at a rate of 10(-6) and mutants resistant to kitasamycin at a rate of 10(-5) to 10(-6) (these were not isolated from strain 13034). In contrast, mutants resistant to minocycline were isolated from strain 13034 only at a rate of 10(-4). Susceptibility testings to antituberculosis drugs were carried out as follows. Bacterial suspensions, 10 mg wet weight/ml, were prepared from ten day-old cultures of the strains growing on Ogawa egg medium slants. Each 0.02 ml-sample of the suspensions was inoculated onto Ogawa egg medium with or without a drug by a spiral loop. The media inoculated were incubated at 37 degrees C for 14 days. Minimal inhibitory concentration was determined as a concentration, on which no membraneous growth could occur. The results are shown in Tables 1 to 3. Cross-resistance relationships were observed only between ethionamide and isoniazid. Ethionamide-resistant mutant strains were resistant to isoniazid.(ABSTRACT TRUNCATED AT 250 WORDS)

Antitubercular Agents↗

Effects of roll mixing with beta-cyclodextrin on enhancing solubilities of water insoluble drugs.

To increase a solubility of water insoluble drugs by the use of roll mixing with beta-cyclodextrin (CyD), equilibrium concentrations of ten medicinal drugs indomethacin, griseofulvin, nifedipine and so on were examined in comparison with those of different mixed systems as kneading with water and physical mixing. The equilibrium concentration of drug in the roll mixed systems was more advantageous than the others excluding the systems of cortisone acetate (CA), and it increased apparently as compared to the solubility of the drugs except for kitasamycin. Almost all of drugs being a crystalline state turned readily to an amorphous form by the procedures of roll mixing with CyD. The amorphous product roll mixed with CA or diethylstilbestrol (DB) changed to the crystalline form due to the inclusion complexes of CyD in X-ray diffraction patterns, when these were exposed on humidifing atmospheres of relative humidity (RH) 97% for 24 hours. The another roll mixed products exhibited the crystalline pattern of the hydrated CyD. Apparent increases in the solubility of drug roll mixed were expected to relate a formation of inclusion compounds or certain molecular interactions between the drug and CyD, which was based upon slight changes in absorption bands on IR spectra.

Chemistry, Pharmaceutical↗

[Effect of rokitamycin on upper gastrointestinal contractile activity. Analysis of side-effects on gastrointestinal tract].

In order to determine whether rokitamycin (RKM), one of the macrolide antibiotics, has any side effects on the gastrointestinal tract, the effect of intraduodenal administration of RKM (1.0, 3.0 and 9.0 mg/kg) on gastrointestinal contractile activity was studied by means of force transducers implanted chronically on the gastric body, gastric antrum, duodenum and upper jejunum in conscious dogs. Erythromycin (EM 0.3, 1.0 and 3.0 mg/kg) and kitasamycin (LM 1.0, 3.0 and 9.0 mg/kg), both macrolide antibiotics, were used as control drugs. RKM, when given at 3.0 mg/kg and 9.0 mg/kg doses, induced segmentation contractions only in the duodenum where it was administered. The duration of the RKM-induced contractions was 7.5 +/- 2.5 minutes for 3.0 mg/kg and 15.8 +/- 3.0 minutes for 9.0 mg/kg, and the contractile force of the contractions was 43 to 82% of the maximum contractile force of the interdigestive contractions in the duodenum. EM, at 0.3 mg/kg, evoked a series of strong contractions quite different from those induced by RKM but similar to the natural interdigestive contractions, and with large doses, dose-dependent long-lasting interdigestive contractions were induced. On the other hand, LM did not stimulate notable gastrointestinal contractile activity even at a 9.0 mg/kg dose. In order to eliminate the possibilities of the contraction being caused by the effect of RKM on the duodenum through the general circulation upon absorption, 3.0 mg/kg RKM was given intravenously. It was found that intravenous injection of RKM did not evoke any contractions attributable to the direct action of RKM in the circulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In vitro sensitivities to antimicrobial drugs of ureaplasmas isolated from the bovine respiratory tract, genital tract and eye.

The sensitivity to 18 antimicrobial drugs was examined for 66 strains of Ureaplasma sp isolated from respiratory tracts of calves suffering from enzootic pneumonia, urinary tracts of bulls and eyes of cows suffering from infectious bovine kerato-conjunctivitis. Furamizole, tiamulin fumarate, erythromycin lactobionate, malidomycin C, doxycycline hydrochloride, kitasamycin tartrate, tylosin tartrate, T-2636C, tetracycline hydrochloride, oxytetracycline hydrochloride, chlortetracycline hydrochloride, oleandomycin phosphate, furazolidone, spiramycin adipate, chloramphenicol and thiophenicol showed strong inhibiting activity on all the test strains. Among them, furamizole, tiamulin fumarate and erythromycin lactobionate were most active. Kanamycin sulphate showed weak activity on all the strains tested. The differences in origin of the test strains did not affect their sensitivity to any of the drugs.

Animals↗