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Regulation of penicillinase synthesis: a mutation in Staphylococcus aureus unlinked to the penicillinase plasmid that reduced penicillinase inducibility.

A mutant of Staphylococcus aureus strain 655 was isolated that is restricted in penicillinase induction. Wild-type plasmids that bear penicillinase determinants could not be fully induced in this mutant, 655par-1; hence, the responsible mutation is not located on the plasmid. Mutant plasmid PI(258)penI443, which produces penicillinase constitutively in wild-type cells, was fully constitutive for penicillinase production when it was harbored by mutant 655par-1. Therefore, the bacterial mutation does not interfere directly with the transcription of the penZ gene or translation of the penicillinase messenger ribonucleic acid. Mutant plasmid PII(147)penI220 was fully inducible in the mutant bacterium, even though the wild-type plasmid PII(147) was only partially inducible in the par-1 mutant. Thus, in the presence of inducer, complementation appears to occur between the product of the par-1 gene and the product of the penI220 gene. These results suggest that the par-1 gene codes for a penicillinase antire-pressor.

Chromosomes, Bacterial↗

Penicillinases of Klebsiella pneumoniae and their phylogenetic relationship to penicillinases mediated by R factors.

On the assumption that the penicillinase determinants on a group of R factors conferring ampicillin resistance have a phylogenetically close relationship to the penicillinase gene of the Klebsiella group, the penicillinases from four strains of K. pneumoniae, GN69, GN1103R(-), GN422, and GN118, were purified 230- to 1,000-fold and compared with the known two R-factor-mediated penicillinases. By gel filtration on Sephadex G-75, the molecular weights were estimated to be 17,400, 18,100, 20,000 and 18,300, respectively, which are slightly lower than those of the R-factor penicillinases. The isoelectric points of the Klebsiella penicillinases were not in agreement with those of the R-factor penicillinases. All the enzymes showed a pH optimum between 6.3 to 7.2 and a temperature optimum of 45 C, and those properties, together with behavior towards inhibitors, were about the same as those in the R-factor penicillinases. The substrate specificity and the Michaelis constants of the Klebsiella penicillinases for penicillins and cephaloridine were broadly similar to those of the R-factor penicillinases, however, some variations were found even among the four penicillinases of K. pneumoniae. The reactivities of the four penicillinases of K. pneumoniae with the antiserum against one R-factor penicillinase were tested, and three of the four Klebsiella penicillinases were found to be indistinguishable immunologically from both R-factor penicillinases. The remaining Klebsiella penicillinase, from GN1103R(-), showed an immunological partial homology with the R-factor penicillinases.

Ampicillin↗

NEW ASSAY FOR PENICILLINASE AND SOME RESULTS ON PENICILLINASE INDUCTION.

Imsande, John (Western Reserve University, Cleveland, Ohio). New assay for penicillinase and some results on penicillinase induction. J. Bacteriol. 89:1322-1327. 1965.-A rapid, sensitive, and reliable assay of penicillinase activity is described. Studies conducted on the induction of penicillinase, with use of the new assay, show that actinomycin D inhibits growth and the induced synthesis of penicillinase in Bacillus cereus 569. These inhibitory effects can be reversed to various degrees by deoxyguanosine, depending upon the time lapse between the addition of the antibiotic and the addition of the deoxynucleoside. Inhibition of growth is reversed more readily than inhibition of penicillinase induction, and it is suggested that actinomycin D may preferentially inhibit the induced synthesis of penicillinase. Studies conducted in an attempt to ascertain the role of penicillin in inducing penicillinase formation in B. cereus 569 suggest that the inducer, penicillin, enhances penicillinase synthesis in some manner in addition to its probable role of inducing the formation of a penicillinase-specific ribonucleic acid messenger.

Bacillus cereus↗

Relationship between penicillinase production and the in-vitro activity of methicillin, oxacillin, cloxacillin, dicloxacillin, flucloxacillin, and cephalothin against strains of Staphylococcus aureus of different phage patterns and penicillinase activity.

A total of 157 strains of Staphylococcus aureus of different phage patterns and penicillinase production were investigated for their susceptibility to methicillin, oxacillin, cloxacillin, dicloxacillin, flucloxacillin and cephalothin by an agar dilution method. Only strains of the 52, 52A, 80, 81 complex had significantly higher IC-50 values than the rest of the strains. No correlation was found between penicillinase production and the IC-50 values. Penicillinase susceptibility divided the antibiotics into two groups: one including methicillin, oxacillin and cephalothin, and the other included dicloxacillin, cloxacillin and flucloxacillin. Nineteen strains of S. aureus which existed in both a penicillinase producing and a penicillinase non-producing form were examined for susceptibility to the six antibiotics. The difference between penicillinase positive and penicillinase negative variants was especially marked for flucloxacillin and cephalothin. Methicillin induction prior to susceptibility testing had only a minor influence on the results. Investigation of the stability of methicillin and the four isoxazolyl penicillins against penicillinase production by 37 strains of S. aureus showed methicillin to be the most stable antibiotic. This was followed by dicloxacillin, cloxacillin, flucloxacillin, and oxacillin in that order. The order of stability was identical and independent of phage pattern and quantitative penicillinase production.

Bacteriophage Typing↗

Constitutive penicillinase formation in Staphylococcus aureus owing to a mutation unlinked to the penicillinase plasmid.

Previously described penicillinase-constitutive mutations in Staphylococcus aureus are caused by genetic lesions in a regulator gene (or genes) on the penicillinase plasmid in close linkage to the structural gene. This report describes a new class (R2(-)) of penicillinase-constitutive mutants of S. aureus unlinked to the plasmid. By transductional analysis, the penicillinase plasmids in these mutants were wild type. Wild-type plasmids transduced into penicillinase-negative (plasmid loss) derivatives of R2(-) mutants produced penicillinase constitutively in amounts comparable to a fully induced culture of the wild-type strain. Penicillinase production in R2(-) mutants was maximal at 30 to 32 C and was much reduced at 40 C.

Enzyme Induction↗

Carbenicillin-hydrolyzing penicillinases of Proteus mirabilis and the PSE-type penicillinases of Pseudomonas aeruginosa.

Three carbenicillin-hydrolyzing penicillinases were found in Proteus mirabilis strains, N-3, N-29, and GN79. The former two strains were isolated in 1978, but strain GN79 was one of our stock cultures isolated in 1965. These penicillinases closely resembled each other, and the PSE-1 and PSE-4 enzymes produced by Pseudomonas aeruginosa, in their substrate profiles and kinetic properties for hydrolyzing various beta-lactams. However, differences were found in their molecular weights and isoelectric points which ranged from 22,000 to 27,000 and from 6.0 to 6.9, respectively. The antiserum against the purified penicillinase of N-29 cross-reacted with the enzyme of N-3 and inhibited its activity by more than 80%. The antiserum also reacted with the PSE-1 and PSE-4 enzymes. The antiserum did not react with the penicillinase from strain GN79 and the PSE-2 and PSE-3 enzymes of P. aeruginosa. Enzyme production in N-3 and N-29 was mediated by R plasmids.

Animals↗

Sensitivity of penicillinase-forming strains of Staphylococcus aureus and of their penicillinase-negative variants to cephaloridine, cephalothin, methicillin, and benzylpenicillin.

Twenty-eight penicillinase-forming cultures of Staphylococcus aureus and their penicillinase-negative variants were examined for resistance to benzylpenicillin, methicillin, cephalothin, and cephaloridine. The results supported the view that cephaloridine was more easily destroyed by staphylococcal penicillinase than was cephalothin. In our tube-dilution tests, the minimum inhibitory concentration (M.I.C.) of cephaloridine for methicillin-sensitive cultures was never as high as some of the values reported by other workers who used apparently comparable methods. This was probably due to small differences in technique. The M.I.C. is an unsatisfactory measure of the antibiotic sensitivity of an organism which produces an enzyme which destroys the antibiotic.Methicillin-resistant strains of Staph. aureus have an intrinsic resistance of ;heterogenous' type also to benzylpenicillin, cephalothin, and cephaloridine.

Cephaloridine↗

Susceptibility of the "penicillinase-resistant" penicillins and cephalosporins to penicillinase of Staphylococcus aureus.

The activities of some semisynthetic penicillins and cephalosporins have been tested against clinical strains of Staphylococcus aureus. The apparent activity in vitro varies with the method of testing used. Determination of MICs using light inocula fails to detect the destructive effect of penicillinase on the antibiotic. This was, however, demonstrated reproducibly by the use of a technique in which a heavy inoculum was pre-incubated for two hours before application of antibiotic to wells. This method of testing probably represents most of the clinical situations in which the drugs are used since both in vitro and in vivo a growing culture is exposed to an antibiotic gradient. Flucloxacillin was inactivated by penicillinase considerably more than either methicillin, cloxacillin, or nafcillin. Cephaloridine was the most vulnerable of the cephalosporins. Cephazolin, cephalothin, and cephalexin were intermediate. Cephradine was the least hydrolysed by staphylococcal penicillinase. It is recommended that the activities of all penicillins and cephalosporins against staphylococci should be tested by diffusion at 37 degrees C with pre-incubation of the culture for two hours at this temperature.

Cephalosporins↗

Antibiotic sensitivity patterns of penicillinase-positive and penicillinase-negative strains of Neisseria gonorrhoeae isolated in Fukuoka, Japan.

Penicillinase production and minimum inhibitory concentrations (MICs) of penicillin G, amoxycillin, doxycycline, and spectinomycin were studied for 155 gonococcal strains isolated in Fukuoka, Japan, between April and December 1981. Of 155 isolates, 25 (16.1%) strains were identified as penicillinase-producing Neisseria gonorrhoeae (PPNG) by a rapid iodometric test. The antibiotic sensitivity of 130 penicillinase-negative (non-PPNG) and 25 PPNG strains was compared. The mean MIC of penicillin G for non-PPNG strains was 0.65 microgram/ml. Twenty-five PPNG strains had MICs of penicillin G between 6.25 and greater than 100 micrograms/ml. The susceptibility to amoxycillin and doxycycline of PPNG strains was significantly lower than that of non-PPNG strains. All the strains proved to be sensitive to spectinomycin. The MICs of four antibiotics for non-PPNG strains were significantly correlated.

Amoxicillin↗

[The enzyme penicillinase and plasmid penicillinase in Staphylococcus aureus strains].

Penicillinase enzyme activities and resistance to some heavy metal ions in 25 S. aureus strains isolated in various hospitals in Ankara were investigated. We also tried to find out whether the controlling genes for penicillinase activity were on the plasmids or not. By the assays that were performed, this enzyme was detected in 22 out of 25 S. aureus strains. All of the strains were found to be resistant to mercury (Hg) and lead (Pb), but only 5 of them were sensitive to cadmium (Cd). We were not able to determine the source of the penicillinase in all strains of staphylococci.

Metals↗

Inhibition of penicillinase by epigallocatechin gallate resulting in restoration of antibacterial activity of penicillin against penicillinase-producing Staphylococcus aureus.

The combination of epigallocatechin gallate (EGCg, a main constituent of tea catechins) with penicillin showed synergism against 21 clinical isolates of penicillinase-producing Staphylococcus aureus. Besides binding directly to peptidoglycan, the inhibition of penicillinase activity by EGCg is responsible for the synergism. EGCg inhibited the penicillinase activity in a dose-dependent fashion, with a 50% inhibitory concentration of 10 microg/ml.

Catechin↗

In vitro antimicrobial susceptibilities of penicillinase-producing and non-penicillinase-producing strains of Neisseria gonorrhoeae isolated in Durban, South Africa.

The in vitro susceptibilities of 22 penicillinase-producing and 32 non-penicillinase-producing strains of Neisseria gonorrhoeae to 13 antimicrobial agents, including the new semisynthetic penicillins and cephalosporins, are reported. Ceftriaxone, ceftazidime, and cefotaxime were the most active agents tested; none of them had an MIC of greater than 0.03 micrograms/ml. Amoxycillin plus clavulanic acid and temocillin also showed good activity against both strains of gonococci.

Anti-Bacterial Agents↗

Antimicrobial susceptibility of non-penicillinase and penicillinase-producing Neisseria gonorrhoeae strains isolated in Tripoli, Libya.

Neisseria gonorrhoeae culture gave a positive result in 42 of 64 male adults with purulent urethral discharge. The majority of the infections were acquired outside Libya. Twenty-seven strains (64.3%) were non-penicillinase producing (NPPNG) and 15 (35.7%) were penicillinase producing (PPNG) by starch paper technique. Antimicrobial susceptibility of the strains to 5 antibiotics was carried out by agar-plate dilution technique. Twenty-three NPPNG strains (54.8%) were susceptible to penicillin with a minimum inhibitory concentration (MIC) of less than or equal to 0.5 micrograms/ml. In 4 strains (9.5%), a high resistance to penicillin (MIC greater than or equal to 16 micrograms/ml) appeared to be chromosomally-mediated (CMRNG). All PPNG strains were resistant to penicillin (MIC greater than or equal to 1 microgram/ml). While resistance to erythromycin (MIC greater than or equal to 1 microgram/ml) and tetracycline (MIC greater than or equal to 1 microgram/ml) was observed in 5 strains, resistance to kanamycin (MIC 32 micrograms/ml) and spectinomycin (MIC 64 micrograms/ml) was present in only one strain. Whereas no significant differences were recorded in MICs of erythromycin, tetracycline, kanamycin and spectinomycin between NPPNG and PPNG strains, one PPNG strain was found to be resistant in vitro to all 5 antibiotics.

Drug Resistance, Microbial↗

Norfloxacin in the treatment of gonorrhea due to penicillinase and non-penicillinase producing Neisseria gonorrheae: a review.

In 12 recent clinical studies, norfloxacin given in various dosage regimens was shown to be highly effective in treating 1,588 gonococcal infections in 1,486 patients. A single oral dose of norfloxacin 800 mg cured 99.2% of 783 patients with penicillinase producing strains of Neisseria gonorrheae (PPNG) and non-penicillinase producing N. gonorrheae (NPPNG) causing urethritis, cervicitis and anorectal infections. A single oral dose of 800 mg of norfloxacin was found to be an effective alternative to a 2 g intramuscular dose of spectinomycin and superior to a 2.5 g oral dose of thiamphenicol for the treatment of PPNG urethritis or cervicitis. Its efficacy in treatment of NPPNG infections was shown to be similar to that of a single oral dose of 3.5 g of ampicillin plus 1 g of probenecid. Mild and transient gastrointestinal and neurological adverse events occurred in 3.3% of patients treated with norfloxacin 800 mg. Norfloxacin 800 mg as a single dose is recommended as an effective and safe single-dose treatment of urethritis, cervicitis and anorectal infections caused by PPNG or NPPNG.

Adolescent↗

Production of penicillinases by certain penicillin-resistant bacteria. (C) factors affecting the activities of penicillinases produced by Staphylococcus aureus, S9.

The activities of penicillinases produced by Staphylococcus aureus, S9 were found to be affected by pH, temperature, substrate concentration and type of penicillin derivative used as a substrate. The optimal activities of penicillinases produced by S. aureus, S9 were obtained at pH 6, at 37 degrees C, 0.5-10 mu/ml penicillin G concentration, and by increasing the enzyme concentration.

Hydrogen-Ion Concentration↗

Comparative in-vitro activity of enoxacin against penicillinase- and non-penicillinase-producing Neisseria gonorrhoeae.

The in-vitro antigonococcal activity of enoxacin was measured by a broth microdilution method. Comparisons were made with five clinically relevant marketed drugs against 23 clinical isolates of Neisseria gonorrhoeae, including several penicillinase producers as well as multiply resistant strains. Results showed that enoxacin possessed potent activity against all organisms tested, with MIC values ranging from less than or equal to 0.013 to 0.05 micrograms/ml.

Anti-Bacterial Agents↗

Susceptibility of penicillinase-producing and non-penicillinase-producing strains of Neisseria gonorrhoeae isolated in Durban, South Africa, to 15 beta-lactam antibiotics.

Using two different inoculum sizes (10(4) and 10(7) organisms), we tested 35 penicillinase-producing (PPNG) and 72 non-PPNG strains of Neisseria gonorrhoeae falling into three categories (penicillin-sensitive, intermediately resistant, and resistant) by the agar-dilution method against 15 beta-lactam antibiotics. With the exception of penicillin, ampicillin, amoxicillin, and cephradine, the isolates were uniformly sensitive to all the other antibiotics tested (including spectinomycin, tetracycline, rosoxacin, sulfamethoxazole-trimethoprim [19:1 ratio], and kanamycin). Ceftriaxone and cefotaxime were the two most active compounds tested; all strains were inhibited at concentrations of 0.007 and 0.015 microgram/ml, respectively. Isolates with intrinsic resistance to penicillin were less susceptible to cefoxitin and cefuroxime than were PPNG strains. The effect of inoculum size varied for the different antibiotics; penicillin and cefaclor showed this effect only with PPNG strains, whereas for ampicillin and amoxicillin it was also seen with non-PPNG strains. Cephamandole showed this effect with both penicillin-sensitive and PPNG strains. In view of the increasing resistance to penicillin, we would recommend that alternative drugs be used as first line therapy for gonorrhea in South Africa.

Anti-Bacterial Agents↗

Comparison of ofloxacin and ceftriaxone in the treatment of uncomplicated gonorrhea caused by penicillinase-producing and non-penicillinase-producing strains.

Eighty-nine patients with uncomplicated gonorrhea, including 31 patients (34.8%) infected with penicillinase-producing strains of Neisseria gonorrhoeae, were treated with oral ofloxacin (single 400-mg dose) or intramuscular ceftriaxone (250-mg dose). All 47 patients who received ofloxacin and 41 of 42 patients who received ceftriaxone were cured.

Adult↗