Induction potency of various beta-lactam derivatives in gram-negative rods

The induction potency of various beta-lactams as well as that of 'nonspecific' inducers such as the media employed or body fluids were studied in gram-negative clinical isolates and in their resistant corresponding counterparts. In all wild-type isolates quite a few beta-lactams (mainly cefoxitin and imipenem) shared the ability to induce the chromosomal beta-lactamase, whereas all beta-lactams - including 6-APS and 7-CPS clavulanic acid, and sulbactam (both beta-lactamase inhibitors), exhibited strong induction potency in their corresponding resistant counterparts. Moreover, all resistant counterparts exhibited the phenomenon of 'nonspecific' induction, whereas the wild-type strains did not. Interestingly, in both Proteus vulgaris 4917 W and Providencia rettgeri 862 W wild-type strains most beta-lactam compounds were potent inducers, thus providing an explanation for resistance against beta-lactamase-unstable compounds. The addition of subinhibitory concentrations of the quinolone compound enoxacin or chloramphenicol did not influence the phenotypic beta-lactamase expression, neither in wild-type strains nor in their resistant counterparts, whereas the addition of clindamycin or gentamicin resulted in a marked decrease of enzyme production in cephalosporinase overproducers, the P. vulgaris and the P. rettgeri isolates.     

Amdinocillin: interaction with other beta-lactam antibiotics for gram-negative bacteria

Amdinocillin was studied alone and in combination with three other beta-lactam antibiotics (aztreonam, cefoperazone, and ceftriaxone) for activity against gram-negative bacilli. These antibiotic combinations failed to show synergy by the checker-board double-dilution test or by killing kinetic studies. However, amdinocillin did show additive killing action when combined with the other beta-lactam antibiotics studied. Amdinocillin failed to induce or inhibit beta-lactamase production in species of Enterobacteriaceae, but with Pseudomonas aeruginosa, beta-lactamase production was induced. It is concluded that the activity of amdinocillin alone or in combination with another beta-lactam antibiotic should not be more effective in the treatment of infections by gram-negative bacilli than just using the beta-lactam antibiotic alone at a higher dose.     

Comparison of antibiotic resistance of gram-negative bacilli with beta-lactam and aminoglycoside antibiotics

In 1981 a comparison of the resistance rates of aminoglycosides, penicillins, cephalosporins, and nalidixic acid was made in species of gram-negative rods between a hospital in Toulouse, France and one in Brooklyn, New York. The results showed similar rates of resistance from both institutions. Both institutions showed high rates of aminoglycoside resistance; high ampicillin resistance among salmonella and Pseudomonas mirabilis in France and shigella in New York City; similar rates of resistance to cephalosporins and finally, markedly different incidences of resistance to naladixic acid. Analysis of medical records over several years indicated a gradual but continuing increase in resistance to aminoglycosides. The majority of such isolates had been isolated from patients in intensive care units. Few differences in the rates of resistance to the cephalosporins were noted between the two institutions, either for the older or newer agents in this group. Further, no increase in resistance was noted to this group of antibiotics in the previous 5 yr.     

In vitro interactions of amikacin and beta-lactam antibiotics against amikacin-resistant gram-negative bacilli

We tested 42 strains of amikacin-resistant gram-negative bacilli with amikacin in combination with six beta-lactam antibiotics using the checkerboard and time-kill curve techniques. Synergism was demonstrated with time-killing curve in 43-68% of the strains tested. Ceftazidime plus amikacin was the most active combination by the checkerboard technique, while amikacin-cefoperazone was the most active combination by the time-killing curve technique against Pseudomonas aeruginosa. Discrepancies were found between the results of the two methods used.     

Inducible type I beta-lactamases of gram-negative bacteria and resistance to beta-lactam antibiotics

Mutants, showing either constitutive (depressed) or non-inducible expression of chromosomally-mediated Type I beta-lactamase were obtained from clinical isolates of Enterobacter cloacae, Ent. aerogenes, Citrobacter freundii, Providencia stuartii, Morganella morganii, Serratia marcescens and Pseudomonas aeruginosa. The wild-type and mutant strains were compared for susceptibility to a range of beta-lactam antibiotics. Derepression of beta-lactamase synthesis generally, but not always, resulted in a marked reduction in susceptibility to the agents tested, including the '3rd generation' cephalosporins. In many cases, the observed resistance would preclude, or severely compromise, the therapeutic efficacy of the drugs. In this context, depressed mutants of Enterobacter spp., Citro. freundii and Ps. aeruginosa could be of primary concern although those of Ser. marcescens, Prov. stuartii and Morg. morganii often exhibited equally high resistance levels to older beta-lactams. Comparison of the susceptibilities of the non-inducible mutants with that of their inducible parents suggested variation in the beta-lactamase inductive potency of different compounds in different organisms. For example, cefoxitin was a powerful inducer in Ent. cloacae, Citro. freundii and one strain of Ps. aeruginosa; similarly cefazolin and cefuroxime were good beta-lactamase inducers in Ser. marcescens and Morg. morganii. Aminothiazolyl-oxime cephalosporins and ureido-penicillins were generally poor inducers. From such comparisons, the contribution of inducible Type I beta-lactamase to resistance phenotype could be ascertained.     

Beta-lactamase-positive strains of Haemophilus influenzae: susceptibility to and inactivation of beta-lactam antibiotics

Susceptibility and time-kill studies were done with low and high inocula of both beta-lactamase-positive and -negative strains of Haemophilus influenzae with cefamandole, ampicillin, cefoperazone, mezlocillin, moxalactam, and ceftriaxone. Bioassay was done to test for antibiotic inactivation by beta-lactamase-positive strains. All six antibiotics were highly active against the low inoculum (10(4) to 10(5) colony-forming units/ml) of beta-lactamase-negative strains; ceftriaxone, moxalactam, and cefoperazone were equally active against the same inoculum concentration of beta-lactamase-positive strains. In contrast, cefamandole, mezlocillin, and ampicillin were less active against the low inoculum of beta-lactamase-positive H influenzae. A marked inoculum effect occurred with the high inoculum (10(7) to 10(8) CFU/ml) with all six antibiotics, regardless of beta-lactamase production. In time-kill studies, marked differences in bacterial killing resulted after low and high inocula. Ampicillin, cefamandole, cefoperazone, and mezlocillin were rapidly inactivated by the high inoculum of beta-lactamase-positive H influenzae.     

Synergic post-antibiotic effect of amikacin in combination with beta-lactam antibiotics on gram-negative bacteria.

The post-antibiotic effect (PAE) of amikacin alone and in combination with ceftazidime, ceftriaxone and piperacillin was studied for two strains each of Pseudomonas aeruginosa and Serratia marcescens using a bioluminescent assay of bacterial ATP. Two models were used for combining beta-lactam antibiotics and amikacin: in one model the cultures were incubated with 32 mg/L of ceftazidime, 128 mg/L of ceftriaxone or 32 mg/L of piperacillin for 1 h. Different concentrations of amikacin (0.5-64 mg/L) were then added. Incubation of the combinations continued for one more hour. The antibiotics were eliminated by dilution. In the second model tested, one strain of S. marcescens was simultaneously exposed to amikacin and a beta-lactam antibiotic for 2 h. The PAEs produced by the drugs in combination were longer than the sum of the individual effects of the drugs when they were used alone. Results were equally good with both models. A synergic PAE was also found with amikacin concentrations close to the MIC in combination with low concentrations of ceftazidime, ceftriaxone and piperacillin.     

Activity of beta-lactamase inhibitors in combination with new beta-lactam antibiotics against resistant gram-negative organisms

Thirty-eight isolates of gram-negative bacilli resistant to four new beta-lactam antibiotics, aztreonam, moxalactam, ceftazidime, and cefoperazone, were tested in the presence of two beta-lactam inhibitors, clavulanic acid and sulbactam. Microorganisms tested included 22 isolates of Pseudomonas species, 5 of Klebsiella species, and 11 of Enterobacter species. A 2- to 10-fold decrease in minimum inhibitory concentration was noted when antibiotics and beta-lactamase inhibitors were combined compared to antibiotics alone. Four strains of Pseudomonas aeruginosa showed increased resistance to the combination of antibiotics and beta-lactamase inhibitors.     

抑制剂增强碳青霉烯类灭活法对革兰阴性杆菌碳青霉烯酶初步分类的评价

目的评价抑制剂增强碳青霉烯类灭活法(ieCIM)对革兰阴性杆菌碳青霉烯酶检测及初步分类的可靠性。方法分别以他唑巴坦和乙二胺四乙酸作为碳青霉烯酶抑制剂,对CIM进行改良。选取临床分离的198株肠杆菌科细菌和35株非发酵菌,采用ieCIM检测碳青霉烯酶并进行初步分类,以PCR方法检测碳青霉烯酶基因作对比。结果 198株肠杆菌科细菌中碳青霉烯酶基因阳性101株,CIM检测阳性99株;碳青霉烯酶基因阴性97株,CIM检测均阴性。35株非发酵菌中碳青霉烯酶基因阳性25株,CIM检测阳性24株;碳青霉烯酶基因阴性10株,CIM检测均阴性。使用ieCIM初步分类碳青霉烯酶,87株产A类酶菌株中检出85株(97.7%),25株产B类酶菌株中检出22株(88.0%),12株产D类酶菌株和2株同时产A、B类酶菌株全部检出,ieCIM检测敏感性为96%,特异性100%。结论 ieCIM与基因检测结果一致性高,适合临床微生物常规工作中对碳青霉烯酶的检测及初步分类。     

Fourth-generation cephalosporins: in vitro activity against nosocomial gram-negative bacilli compared with beta-lactam antibiotics and ciprofloxacin

The in vitro activity of two new expanded spectrum fourth-generation cephalosporins, cefepime and cefpirome, was compared with that of five antibacterial agents, ceftazidime, cefoperazone, cefotaxime, imipenem, and ciprofloxacin, that are commonly used in the treatment of serious infections caused by aerobic gram-negative bacteria. The agar dilution method described by the US National Committee for Clinical Laboratory Standards was used to determine the minimum inhibitory concentrations of antibiotics tested. Three hundred and two clinical isolates, representing a cross-section of Klebsiella and Enterobacter species and Pseudomonas aeruginosa were tested. The most potent beta-lactams were imipenem, cefepime, and cefpirome, which demonstrated significant activity against the majority of strains in all three genera of bacteria, as did ciprofloxacin. Ceftazidime was active against P. aeruginosa, but was less potent against Klebsiella and Enterobacter species. Cefoperazone and cefotaxime were less active than ceftazidime against P. aeruginosa. Cefepime had slightly greater activity than cefpirome against the gram-negative bacteria tested. These data indicate that cefepime and cefpirome are highly active against many frequently resistant nosocomial bacterial strains that are traditionally responsible for difficult-to-treat infections.     

Comparable evaluation of orally active beta-lactam compounds in ampicillin-resistant gram-positive and gram-negative rods: role of beta-lactamases on resistance

The antibacterial activity of the recently developed cephems cefixime and cefetamet-pivoxyl was evaluated in 408 gram-positive and gram-negative rods, all isolated recently from clinical specimens, and compared to that of other orally active agents such as ampicillin, amoxycillin + clavulanic acid, cefaclor, cefuroxime-axetil and to ceftriaxone. With regard to ampicillin-resistant Enterobacteriaceae ceftriaxone proved to be the most active agent, followed by cefixime and cefetamet, whereas cefuroxime was less active. Cefaclor and amoxycillin + claculanic acid were active against ampicillin-resistant Escherichia coli, Klebsiella pneumoniae, and Proteus ssp. isolates. All beta-lactam compounds exhibited poor activity against Acinetobacter anitratus isolates, but were highly active against Haemophilus influenzae with the exception of cefaclor. Both cefixime and cefetamet were poorly active against Staphylococcus aureus, but highly active against beta-hemolytic streptococci. Moreover, both compounds remained unaffected by the production of plasmid-mediated beta-lactamases such as the TEM or OXA enzymes. Resistance to both agents was observed in Enterobacteriaceae that produced large amounts of chromosomally mediated enzymes; their affinity to the class I enzyme from Enterobacter cloacae was somewhat lower than that of other third-generation cephalosporins. However, in contrast with these agents breakdown of cefixime and cefetamet by a class IIIa enzyme form Proteus vulgaris was marginal. In methicillin-resistant S. aureus isolates there was a complete cross-resistance between all beta-lactam compounds included in this study.     

The comparative activity of eleven aminocyclitol antibiotics against 773 aerobic gram-negative rods and staphylococci isolated from infected hospitalized patients

The minimum inhibitory concentrations (MICs) of 11 aminocyclitol antibiotics were determined for 773 recent clinical isolates from infected hospital patients, most of whom were immunocompromised. There was a bias towards aminoglycoside-resistant organisms, and 21% of Pseudomonas aeruginosa isolates were resistant to gentamicin. The antibiotics included the natural agents gentamicin, tobramycin, sissomicin and kanamycin and the semi-synthetic compounds amikacin, netilmicin and dibekacin with some newer agents, including O-demethyl fortimicin, Hapa gentamicin B and 5-epi-sissomicin, not available commercially. Tobramycin, sissomicin and gentamicin had similar spectra with tobramycin more active against Ps. aeruginosa, sissomicin more active against proteus and gentamicin more active against serratia. The differences in spectrum between amikacin and netilmicin were marginal, reflecting the relatively low prevalence of acetyltransferase-producing organisms in our collection. Hapa-gentamicin B was the most active aminoglycoside against staphylococci and indole-positive proteus, netilmicin against Escherichia coli and 5-epi-sissomicin against Ps. aeruginosa. Overall 5-epi-sissomicin was the most active aminoglycoside tested. Both Hapa-gentamicin B and 5-epi-sissomicin have potentially valuable antibacterial spectra which merit clinical studies.     

In vitro activity of amdinocillin in combination with other beta-lactam antibiotics against aminoglycoside-susceptible and resistant gram-negative bacteria

Amdinocillin alone and in combination with other beta-lactam antibiotics was tested for in vitro activity against aminoglycoside-susceptible and resistant gram-negative bacteria. Amdinocillin alone or in combination with ampicillin, ticarcillin, piperacillin, cefazolin, cefoxitin, and cefamandole had little to no activity against aminoglycoside-resistant E. coli, E. cloacae, K. pneumoniae, and S. marcescens. There was better activity with aminoglycoside-susceptible organisms, however, Overall, there was significantly more antagonism of amdinocillin combinations when tested with aminoglycoside-resistant organisms than with aminoglycoside-susceptible strains.     

Inactivation of beta-lactam antibiotics by Legionella pneumophila.

Beta-lactam-inactivating activity has been found in all sero-groups of Legionella pneumophila. The beta-lactamase activity could be detected in intact cells and released by ethylenediaminetetraacetic acid treatment, indicating that it is located in the periplasmic space. The enzyme acted primarily as a cephalosporinase hydrolyzing cefamandole, cephalothin, cephaloridine, and also penicillin G and ampicillin. Cefoxitin and cefuroxime were not hydrolyzed. Clavulanic acid and CP-45,899, beta-lactamase inhibitors, prevented the hydrolysis of cephalosporins and penicillins. The beta-lactamase activity appears to be different from that found in Enterobacteriaceae and Pseudomonas.     

In vitro synergistic activity of some chinolinic compounds combined with beta-lactam antibiotics against gram-positive and gram-negative clinical isolates

The antimicrobial activities of nalidixic acid-cephalexin (ratio 1:1) and cinoxacin-cefadroxil (ratio 1:2) combinations have been evaluated against 396 clinical isolates; many of them were nalidixic acid- or cinoxacin-resistant organisms (MIC greater than or equal to 100 micrograms/ml). We have also tested the nalidixic acid-amoxicillin combination (ratio 1:1) against 225 amoxicillin-resistant bacterial strains (MIC greater than or equal to 800 micrograms/ml). Synergy was found for 62-70% of the Enterobacteriaceae and nonfermenter bacilli tested and for 85-92% of the gram-positive bacterial strains. The 225 clinical isolates resistant to amoxicillin (MIC greater than or equal to 800 micrograms/ml) were synergistically inhibited by the nalidixic acid-amoxicillin combination.     

Pharmacokinetic evaluation of beta-lactam antibiotics

A simple method is proposed to predict free tissue concentrations of beta-lactam antibiotics from their plasma concentrations after iv bolus injection. During the elimination phase these free tissue concentrations exceed corresponding free plasma concentrations by a constant compound specific factor. This factor can be calculated from the different volumes of distribution (Vc, Vdss and Vdarea) and the plasma protein binding. Calculation of free tissue concentrations allows more secure interpretation of pharmacokinetic data with respect to in-vitro MICs for the comparison of different antibiotics or of the same antibiotic in different patient populations.