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Inducible and constitutive resistance to macrolide antibiotics and lincomycin in clinically isolated strains of Streptococcus pyogenes.

STUDIES ON ERYTHROMYCIN RESISTANCE IN STRAINS OF GROUP A STREPTOCOCCI INDICATED THAT THEY WERE COMPRISED OF TWO TYPES: (i) an inducible, resistant type (IR strains) was seen, which manifested immediate logarithmic growth in media containing high concentrations of the drug only after brief previous exposure (induction period) of the organisms to subinhibitory concentrations of erythromycin, and (ii) a constitutive, resistant type (CR strains) which demonstrated, without prior drug exposure, continued logarithmic growth in media containing high concentrations of erythromycin. Subinhibitory concentrations of either chloramphenicol or puromycin, when added to IR strains prior to induction, interfered with their induction by erythromycin. Exposure of CR strains to chloramphenicol did not visibly affect the subsequent growth curve of these strains in media containing high concentrations of erythromycin. In IR strains, resistance to other macrolide antibiotics (oleandomycin, spiramycin, carbomycin, magnamycin) and to lincomycin also was inducible in nature. There was cross-inducibility between erythromycin, other macrolide antibiotics, and lincomycin. CR strains were constitutively resistant to these antibiotics.

Anti-Bacterial Agents↗

Identification of functional amino acids in the macrolide 2'-phosphotransferase II.

Macrolide 2'-phosphotransferase [MPH(2')] transfers the gamma phosphate of ATP to the 2'-OH group of macrolide antibiotics. The role of aspartic acids in the putative ATP-binding site of MPH(2')II was investigated through the substitution of alanine for aspartate by site-directed mutagenesis. D200A, D209A, D219A, and D231A mutant strains were unable to inactivate the substrate oleandomycin, while a D227A mutant retained 7% of the activity of the original enzyme.

Adenosine Triphosphate↗

Evaluation and modifications of media for enumeration of Clostridium perfringens.

The suitability of the Shahidi-Ferguson perfringens, TSC (tryptose-sulfite-cycloserine), and oleandomycin-polymyxin-sulfadiazine perfringens agars for presumptive enumeration of Clostridium perfringens was tested. Of these, the TSC agar was the most satisfactory. The TSC agar method was improved by eliminating the egg yolk and using pour plates. The modified method allowed quantitative recoveries of each of 71 C. perfringens strains tested and is recommended. For confirmation of C. perfringens, the nitrite test in nitrate motility agar was unreliable, particularly after storage of the medium for a few days. In contrast, positive nitrite reactions were obtained consistently when nitrate motility agar was supplemented with glycerol and galactose.

Agar↗

Inhibition of protein synthesis by polypeptide antibiotics. I. Inhibition in intact bacteria.

Ennis, Herbert L. (St. Jude Children's Research Hospital, Memphis, Tenn.). Inhibition of protein synthesis by polypeptide antibiotics. I. Inhibition in intact bacteria. J. Bacteriol. 90:1102-1108. 1965.-The mechanism of inhibition of growth of cells by the polypeptide antibiotics of the PA 114, vernamycin, and streptogramin complexes was studied. This inhibition apparently was due to the selective inhibition of protein synthesis by these antibiotics. Ribonucleic acid synthesis was unaffected by concentrations of the antibiotics which completely inhibited protein synthesis. Deoxyribonucleic acid synthesis was slightly inhibited. These antibiotics are composed of a number of components. Mixtures of equal amounts of PA 114 A and PA 114 B or vernamycin A and Balpha were more active in stopping protein synthesis in intact cells than each of the components of the antibiotic complex alone. Mutants resistant to one of the antibiotics were resistant to all of the group and, in addition, were resistant to erythromycin and oleandomycin.

Anti-Bacterial Agents↗

Role of substrate mycelium in colony development in Streptomyces.

Cellophane cultivation techniques have been proven to be useful for the study of colony growth in Streptomyces. Results obtained by this procedure indicate that, in S. antibioticus, substrate mycelium was a nutrient support for aerial mycelium growth. Oleandomycin synthesis starts before aerial mycelium formation and may play an important role during colony growth.

Autoradiography↗

Antibiotics commonly used to treat mastitis and respiratory burst of bovine polymorphonuclear leukocytes.

The in vitro effects of six doses (2 x 10(-3) to 2 x 10(-8) M) of antimicrobial drugs that are frequently used in udder infusions on the capacity of bovine blood polymorphonuclear neutrophilic leukocytes to generate reactive oxygen species were studied by the measurement of luminol-dependent chemiluminescence after stimulation with phorbol 12-myristate 13-acetate. All drugs, except cloxacillin, significantly decreased chemiluminescence at the highest dose. Doxycyline induced the most severe inhibition, followed by neomycin and dihydrostreptomycin. The effect of ampicillin was due to the scavenging of reactive oxygen species and interactions with luminol. The inhibition observed with oleandomycin, neomycin, lincomycin, and dihydrostreptomycin was not due to direct effects on the production of oxidative metabolites but rather to interference with other components involved in the production of light, such as interference with the interaction between luminol and the myeloper-oxidase-H2O2-halide system. The deleterious effects of doxycycline can be explained by several factors: decreased production of superoxide, yellow color, the scavenging of reactive oxygen species, and Ca2+ chelating effect. In conclusion, the results of this study show that antibiotics may affect neutrophil function at concentrations that are reached in the mammary gland after local and repeated administration.

Ampicillin↗

Enzymatic phosphorylation of macrolide antibiotics.

Five macrolide antibiotics (erythromycin A, 1; oleandomycin, 3a; tylosin, 4a; spiramycins, 5a; leucomycin A3, 6a) have been phosphorylated enzymatically using cell-free extracts derived from Streptomyces coelicolor UC 5240. The necessary cofactors and the rates of the conversion have been determined.

Anti-Bacterial Agents↗

Agar well technique for antibiotics in animal feeds.

The agar well technique compares favorably with the cylinder plate assay in accuracy, sensitivity, and precision. It is more flexible and more rugged, and growth of seed organism is not inhibited. The wells are precision cut with Grafar gel punch assembly with sets (6) of 10, 7, 5, 4, and 3 mm cutters. The wells are easily and rapidly filled with short, disposable Pasteur pipets fitted with rubber bulbs. The smaller wells are filled with capillary pipets. The diameter of the well (independent of volume) appears to be a function of concentration. For every decrease in the diameter size of the well, concentration can be increased, at least to the next higher level of the standard response line. Extracts of chlortetracycline containing as much as 3.2 mu-g/ml can be analyzed if the 3 mm wells are used and with no sacrifice in accuracy or precision. This works especially well for antibiotic premixes. The technique has been used successfully for penicillin, streptomycin, chlortetracycline, oxytetracycline, oleandomycin, and tylosin.

Agar↗

Resistance of Mycoplasma pneumoniae to macrolide and analogous antibiotics.

1. The susceptibility of M. pneumoniae to antibiotics can be determined by the microtiter method. The adequate technique requires that the final volume of broth medium in a well is 0.2 ml and that the dilution is made after the parent solution of antibiotic in the test tube is dropped into a well every fifth wells. 2. M. pneumoniae was cultured on agar media containing two-fold concentrations of macrolide and analogous antibiotics, and the following results were obtained. 1) The growth of eight strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of thet strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of thet strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of the colony grown as an average of 0.5 to eight on agar media containing erythromycin in four strains was 0.1 to 1.6 micrograms/ml in some colonies, and 400 to 800 micrograms/ml in most colonies. The results disclosed that the broth culture contains a small number of mycoplasma cells with a definite, high degree of resistance to the antibiotics, but no cells with intermediate degrees of resistance. 3) The FH strain was made resistant to erythromycin, oleandomycin, midecamycin, acetylspiramycin, leucomycin, josamycin, tylosin, lincomycin, or clindamycin by subculture in broth medium from the colony grown at the highest concentrations of each of the antibiotics in agar media. The degree of the resistance developed was 16 to 128,000 in the MIC radio and showed high values of MIC in most strains. The resistance developed was not lost by subculturing the resistant strain in broth medium without antibiotic. 4) The FH strain made resistant to the antibiotics had cross resistance to other macrolides. Strains resistant to some of the antibiotics had cross resistance to lincomycin and clindamycin, and strains resistant to others did not. Some strains made resistant to macrolides with cross resistance to lincomycin and clindaymycin and strains made resistant to lincomycin or clindamycin had no cross resistance to vernamycin B alpha, while all the resistant strains without cross resistance to lincomycin and clindamycin had cross resistance to vernamycin B alpha. No strain had cross resistance to vernamycin A...

Agar↗

[Macrolide antibiotics for oral intake in the treatment and prevention of suppurative surgical infection].

The authors studied the clinical efficacy of antibiotics of the macrolides group--erythromycin, oleandomycin, triacetylalleandomycin and leukomycin--in complex treatment of 280 patients with suppurative surgical infection caused by a gram-positive, mainly staphylococcal, microbial flora. Various medicinal forms were used: tablets, capsules, granules, syrup. Efficacy of treatment comprised 70-78 per cent. Tolerance of antibiotics was good in the majority of patients.

Administration, Oral↗

[Possible synergic action of some drug combinations against Mycoplasma gallisepticum].

Yamamoto and Adler's method (1956), making use of the modified medium of Edward, was employed to determine the minimal concentrations of gentamicin (GMC), erytrhomycin (ERM), oleandomycin (OMC) spectinomycin (SPM), tetracycline (TC), tylosin (TLS), and chloramphenicol (CA), all of them showing bactericidal effects on Mycoplasma gallisepticum-S6, applied alone or in combination. In vitro studies showed that relatively most effective were TLS, GMC, and SPM, when used alone. Combining the antibiotics by two in the most cases led to a pronounced synergic effect. Comparatively most effective were the combinations of TLS+ERM; TLS+TC; CA+TC; CA+OMC; TLS+CA; TLS+OMC; SPM+TC; and SPM+CA. Lowest absolute concentrations of the individual components showed the combinations of TLS+GMC; TLS+TC; TLS+ERM; TLS+CA; TLS+SPM; TLS+OMC; SPM+TC; and SPM+CA.

Anti-Bacterial Agents↗

[Assessment of the sensitivity of Neisseria meningitidis strains to benzylpenicillin using the disc diffusion method].

The sensitivity of 235 N. meningitidis strains to 5 antibiotics was estimated by the diameter of growth inhibition zones according to the criteria recommended by a Laboratory (Marseilles, France) collaborating with the WHO. All the strains proved to be sensitive to benzylpenicillin when disks containing 10 and 2 IU of the antibiotic were used. The strains were also shown to be sensitive to chloramphenicol and tetracycline. 95.7 and 7.7 per cent of the strains were sensitive to rifampicin and oleandomycin, respectively. When the strain sensitivity was assayed with the disks containing 10 and 2 IU of benzylpenicillin by the more severe criteria recommended by J. Saez-Nieto et al., significant changes were detected: meningococci with relative resistance to benzylpenicillin were detected in various regions of this country and the number of such strains was found to have a tendency to slightly increase.

Anti-Bacterial Agents↗

[Olemorphocycline treatment of calculous and noncalculous pyelonephritis].

40 patients suffering from calculous and non-calculous pyelonephrits were treated with olemorphocycline administered intravenously in a dose of 250 00 Units 2 times a day for 5--7 days. The clinical effect was observed in 62.5 per cent of the cases. In 54.7 per cent of the cases the isolates were resistant to oleandomycin and morphocycline. The kidney tissue and urine from the kidney pelvis taken during surgical operations of 20 patients were plated out. Staphylocci were most often detected in the platings of the kidney tissue. The microflora of the urinary bladder before the treatment and that of the urine from the kidney pelvis and the kidney tissue taken during the surgical operation was mainly ident

Adolescent↗

[Sensitivity of Streptomyces globisporus 3-1--a highly active producer of landomycin E to its own and the other polyketide antibiotics].

The strain Streptomyces globisporus 3-1--a highly active producer of a new anticarcinogenic antibiotic landomycin E from the group of angucyclins has been studied for its resistance to its own and known polyketide antibiotics--tetracycline, oleandomycin and doxorubicine as well as to aminoglycoside antibiotic streptomycin. The survival of mycelium of the strain 3-1 was established to depend on antibiotics concentration. Four equal resistances of streptomycete to antibiotics within the concentration of 5-30 micrograms/ml have been shown. The survival of the strain 3-1 mycelium fragments at 5 micrograms/ml of antibiotic is for tetracycline--1.8, for olcandomycin--0.7; doxorubicin and landomycin E--(-0.7) and streptomycin--(-4.6) l g%. As to their intensity of the lethal effect on the strain 3.1 antibiotics show the following descending succession: streptomycin, landomycin E, doxorubicin, olcandomycin and tetracyclin. High sensitivity of the strain 3-1 to the lethal effect of its own antibiotic may be explained by high permeability of mycelium fragments to landomycin E as compared to the intact mycelium.

Aminoglycosides↗

[Effect of the producer type on the nature of antibiotic fermentation].

Fermentation processes in production of bacitracin, a polypeptide antibiotic by Bacillus licheniformis, and oleandomycin, a macrolide antibiotic by Streptomyces antibioticus, were studied comparatively. It was shown that the antibiotic-producing actinomycete was characterized by a prolonged phase of growth retardation. The highest efficiency of the control actions was observed at the beginning of the fermentation. They were aimed at intensifying the substrate usage during the growth phase and activation of cell metabolism. Controlled cultivation of the Bacillus representative was based on its capacity of achieving the maximum growth rate possible under the certain conditions. Therefore, an increase in the quantity of the synthesized antibiotic was due, under such conditions, to inhibition of the culture growth by various means including lower mass exchange intensity.

Bacillus↗

[Microcalorimetric Investigations, regarding the antibacterial efficiency of chemotherapeutics (author's transl)].

An investigation method is described by which microcalorimetry and continuous density measurement are used to show the antibacterial efficiency of chemotherapeutics. Oleandomycin, erythromycin, both macrolide antibiotics, and doxycycline, a tetracycline, blockers of the protein synthesis, have the same effect on the microcalorimetric curve of a staphylococcus strain. Among the aminoglycosides tobramycin and gentamicin influenced the microcalorimetric activity of a growing E. coli strain in the same way, spectinomycin produced different curves. Penicillins interfere with the synthesis of the cell membrane by blocking the enzyme transpeptidase. Epicillin, penicillin, ampicillin, oxacillin, carbenicillin, and azlocillin influence the microcalorimetric activity of staphylococci in a typical way which is different from that of the blockers of the protein synthesis.

Aminoglycosides↗

[Sensitivity of m. kansasii strains to various antibiotics and chemotherapeutics in vitro and in vivo (author's transl)].

Sensitivity of 40 strains of M. kansasii (var. luciflava) to various antibiotics and chemotherapeutics in vitro and in vivo was examined. The strains showed considerable individual variability in their susceptibility in vitro, most were more resistant to isoniazid and PAS and about one half of the investigated strains to streptomycin, too, if correlated to "wild" strains of M. tuberculosis. Susceptibility to ethionamide, thiosemicarbazone (Conteben), viomycin, kanamycin and cycloserine appears to be the same as in the case of M. tuberculosis strains, but the resistance to pyrazinamide is higher. Good effect has been observed in vitro with rifampicin, ethambutol, capreomycin and phenazine derivative B 663. Oleandomycin and gentamycin are ineffectual in the dosages of drugs we used. The activity of most of these drugs was observed in experimental infections of white mice--in mono-therapy and in various combinations-provoked by different strains of M. kansasii. The main criterion of the effectivity of the treatment was the enumeration of viable mycobacterial units in the lungs of the mice in ten-days intervals; in one part of the experiments the histological examination of the various organs of the mice was performed. The drugs of the I. and II. line of antituberculotics were ineffective or showed only a slight effect. A very good effect of rifampicin, ethambutol and phenazine derivative B 663 in monotherapy and in multiply drug therapy was observed.

Animals↗

[Tolerance for tetraolean i.m. and the blood levels in agricultural animals].

Studied was a drug formula of tetraolean for i/m application, containing tetracycline hydrochloride and oleandomycin phosphate in a 2:1 ratio, produced by the Antibiotic Works, Razgrad, and released in flasks of 500 mg and 1.0 g. It was followed up with regard to its tolerable serum concentrations for rabbits, calves, pigs, and horses. It was found that in therapeutic amounts tetraolean was well tolerated and did not lead to local and general disturbances. In higher doses it was found to form transient oedema in the site of injection. Intramuscularly, tetraolean was well absorbed in the site of application, and, according to the animal species, it developed proper plasmal therapeutic concentrations at the following rates: calves--20 mg/kg body mass at 12-hour intervals; pigs--10 mg/kg at 6-hr intervals, or 20 mg/kg at 12-hr intervals; horses--10 mg/kg at 12-hr intervals or 20 mg/kg at 24-hr intervals.

Animals↗