In vitro and in vivo studies of three antibiotic combinations against gram-negative bacteria and Staphylococcus aureus.

The activities of azlocillin, cefotaxime, and amikacin alone and in combination were evaluated in in vitro checkerboard studies, in infected neutropenic mice, and in human volunteers. The combination of cefotaxime plus amikacin was more synergistic in vitro than the others against the Enterobacteriaceae tested, and the combination of azlocillin plus amikacin was more synergistic against Pseudomonas aeruginosa and Staphylococcus aureus. Survival of neutropenic mice infected with Escherichia coli and Klebsiella pneumoniae, respectively, was greater with azlocillin plus amikacin (24 of 40 and 11 of 40) and with cefotaxime plus amikacin (21 of 40 and 17 of 40) than with azlocillin plus cefotaxime (22 of 40 and 3 of 40; P less than 0.05). Median serum bactericidal activity in volunteers receiving these antibiotics alone and in combination was greater than or equal to 1:8 with most agents and with all combinations tested against 10 strains each of E. coli, K. pneumoniae, P. aeruginosa, and S. aureus. These data suggest that clinical trials with combinations of azlocillin or cefotaxime plus amikacin deserve further study in febrile neutropenic patients.     

Paradox between the responses of Escherichia coli K1 to ampicillin and chloramphenicol in vitro and in vivo

We evaluated the activity of ampicillin and chloramphenicol in vitro and in vivo against an Escherichia coli K1 strain. In vitro, the strain was relatively susceptible to both antibiotics (MIC and MBC of ampicillin, 2 and 4 micrograms/ml; MIC and MBC of chloramphenicol, 4 and 64 micrograms/ml). Checkerboard determinations of MBCs of drug combinations were consistent with antibiotic antagonism. Killing curves with concentrations of antibiotics similar to in vivo levels in blood and cerebrospinal fluid of infected rats indicated antagonism within the first 4 h and an indifferent effect of the combination at 24 h. Paradoxically, the combination was significantly more effective than ampicillin or chloramphenicol alone in vivo in infant rats. This was shown by (i) more rapid bacterial clearance from the blood and cerebrospinal fluid, (ii) a decreased incidence of meningitis in bacteremic animals, and (iii) improved survival. These findings illustrate a divergence between the effects of ampicillin and chloramphenicol against E. coli in vitro and in vivo and suggest that this combination is an effective synergistic regimen in this experimental model of E. coli bacteremia and meningitis.     

Synergism at clinically attainable concentrations of aminoglycoside and beta-lactam antibiotics.

We evaluated the in vitro synergistic activity at clinically attainable concentrations of combinations of aminoglycoside and beta-lactam antibiotics against 30 gentamicin-resistant clinical isolates of gram-negative bacilli. All 56 pairs of 4 aminoglycosides and 14 beta-lactams were evaluated. Combinations with amikacin demonstrated inhibitory synergistic activity in 29% of the assays, as compared with 22% for netilmicin (P = 0.018), 17% for gentamicin (P less than 0.001), and 13% for tobramycin (P less than 0.001). Among the beta-lactams, combinations with cefoperazone, ceftriaxone, or cefpiramide (SM-1652) demonstrated inhibitory synergistic activity most often (39, 38, and 35% of the assays, respectively) and with ceforanide, cefsulodin, and imipenem least often (less than or equal to 8% each). The most active combination was amikacin and ceftriaxone, with which 67% of the assays demonstrated inhibitory synergism. Isolates with high-level resistance to either antibiotic in a combination were unlikely to be inhibited synergistically by the combination. Further, combinations generally demonstrated little synergistic activity against isolates highly susceptible to beta-lactams.     

Bactericidal activity of beta-lactams and amikacin against Haemophilus influenzae: effect on endotoxin release.

Ampicillin or cefotaxime, alone or in combination with amikacin, were tested at levels achievable in CSF for bactericidal activity against eight clinical isolates of Haemophilus influenzae serotype b. Endotoxin release was determined by the limulus amoebocyte lysate test and by macrophage tumour necrosis factor production for each beta-lactam antibiotic, alone and in combination with amikacin. Accelerated killing was observed when amikacin was added to ampicillin or cefotaxime; however, the additional antibiotic-induced bacterial lysis observed after the addition of amikacin to beta-lactam antibiotics was not associated with an increase in endotoxin release.     

Antibacterial activity of tobramycin against gram-negative bacteria and the combination of ampicillin/tobramycin against E. coli.

The antibacterial activity of tobramycin, gentamicin, erythromycin, cloxacillin, kanamycin, cephalexin, penicillin, carbenicillin and polymyxin were compared against 303 clinical bacterial isolates from a pediatric hospital patient population. Standard disk diffusion and agar-dilution methods were employed. Significant activity was demonstrated for tobramycin against pseudomonas, Klebsiella, Escherichia coli and both Staphylococcus aureus and albus; Tobramycin was significantly more active against Pseudomonas than gentamicin or the other antibiotics testedmcomparable activity to gentamicin was present for the other types of bacteria; Cross-resistance was not encountered between tobramycin and gentamicin. 30 isolates of E. coli were tested against the combination of tobramycin and ampicillin by the growth-curve method. Synergism was demonstrated in 4 isolates, antagonism in 1 and an additive effect in 25. A bactericidal effect was present at 24h against 17 isolates with tobramycin alone and against 25 isolates when combined with ampicillin. These results provide in vitro rationale for the consideration of tobramycin for clinical use in patients with Psuedomonas infections for the combination of ampicillin and tobramycin for the treatment of selected E.coli infections.     

Synergic post-antibiotic effect of mecillinam, in combination with other beta-lactam antibiotics in relation to morphology and initial killing.

The synergic in-vitro post-antibiotic effect (PAE) of mecillinam, in combination with either ampicillin, aztreonam, ceftazidime or piperacillin, on a reference strain of Escherichia coli was evaluated by bioluminescence assay of bacterial ATP. Ampicillin, ceftazidime and mecillinam alone induced a concentration dependent PAE (greater than 3 h) on E. coli, whereas aztreonam and piperacillin alone induced a short (less than 1 h) non-dose dependent PAE. At most concentrations, the combination of mecillinam and ampicillin, aztreonam, ceftazidime or piperacillin induced longer PAEs on E. coli than the sum of the individual antibiotics' PAEs. Long PAEs were seen concomitantly with the presence of spheroplasts. In addition to the synergistic PAE, the decrease in colony counts and changes in ATP values after a 2 h exposure to mecillinam, in combination with the other beta-lactam antibiotics, were more prominent than the respective values after exposure to the individual antibiotics. The change in ATP was generally less pronounced than the decrease in colony counts. This could be due to lysis of spheroplasts on agar plates, leading to an over-estimation of the initial killing when assayed by viable counting. Mecillinam, which induced long PAEs on E. coli at almost all concentrations in this study, has a high affinity for penicillin binding protein 2 (PBP 2) and induced spheroplast formation at all concentrations. However, mechanisms other than the affinity for PBP 2 and spheroplast formation are involved in the PAE of beta-lactam antibiotics on Gram-negative bacteria; since the PAE was prolonged when mecillinam was combined with ampicillin, aztreonam, ceftazidime or piperacillin, which bind preferentially to PBP 1 and 3.     

Activity of Newer Aminoglycosides and Carbenicillin, Alone and in Combination, Against Gentamicin-Resistant Pseudomonas aeruginosa

The in vitro activity of the aminoglycoside antibiotics tobramycin, sisomicin, amikacin, gentamicin, and netilmicin (SCH 20569) were compared against 26 gentamicin-resistant isolates of Pseudomonas aeruginosa cultured from hospitalized children. Tobramycin had the greatest activity on a weight basis, followed by sisomicin, gentamicin, amikacin, and netilmicin. All isolates were resistant to achievable concentrations of netilmicin and gentamicin, but 23% were inhibited by achievable concentrations of tobramycin, 8% by amikacin, and 4% by sisomicin. The combinations carbenicillin/tobramycin, carbenicillin/sisomicin, and carbenicillin/amikacin were synergistic for 92% of strains; antagonism was not encountered. These in vitro results suggest that tobramycin, sisomicin, or amikacin in combination with carbenicillin would be the safest initial regimen in the therapy of gentamicin-resistant Pseudomonas infections pending susceptibility studies.     

Effect of recombinant human activated protein C on the bactericidal activity of human monocytes and modulation of pro-inflammatory cytokines in the presence of antimicrobial agents

OBJECTIVES: To determine the effects of recombinant human activated protein C (rhAPC) on the antimicrobial activity and cytokine production of normal human monocyte-derived macrophages (MDMs) in the presence and absence of Escherichia coli infection, with and without treatment with levofloxacin or ampicillin. METHODS: MDM monolayers were infected with E. coli ATCC 25922 and treated with levofloxacin or ampicillin in the presence or absence of rhAPC. Antimicrobial activity and cytokine (TNF-alpha, IL-1beta, IL-6 and IL-8) concentrations in the supernatants were measured. RESULTS: When low concentrations of levofloxacin were used, a therapeutic concentration of rhAPC enhanced intracellular antibacterial activity at all time points. With ampicillin, antibacterial activity increased, was unaffected or diminished depending upon the drug concentration and assay time. Without antibiotics, rhAPC had no antibacterial effect. E. coli caused cytokine production to increase many fold. This increase was significantly greater with antibiotics (P < 0.01). Without antibiotics, rhAPC decreased production of TNF-alpha, IL-1beta and IL-6, but not IL-8. At high levofloxacin concentrations, rhAPC was associated with further increases in the concentrations of these cytokines. Cytokine concentrations at 24 h were unaffected by rhAPC in the presence of ampicillin and E. coli. CONCLUSIONS: rhAPC can affect the bactericidal activity and cytokine production of human MDM in the presence of infection and antibiotic therapy. Importantly, factors such as type and concentration of antibiotics, presence of bacteria and timing must be taken into consideration when evaluating cytokine data from septic patients.     

Synergistic Activity of Trimethoprim and Amikacin Against Gram-Negative Bacilli

The in vitro effect of trimethoprim on the inhibitory and bactericidal activity of amikacin against 20 strains each of Klebsiella pneumoniae and Serratia marcescens, 15 strains of Escherichia coli, and 10 strains of Pseudomonas aeruginosa was examined by the checkerboard technique in microtiter plates. Trimethoprim had a synergistic effect on the inhibitory and bactericidal activity of amikacin against the majority of non-pseudomonas strains tested. The mean +/- standard deviation fractional inhibitory concentration indexes were 0.59 +/- 0.19 for the Klebsiella strains, 0.48 +/- 0.18 for the Serratia strains, and 0.60 +/- 0.22 for the E. coli strains tested. Respective mean +/- standard deviation fractional bactericidal concentration indexes for these organisms were 0.55 +/- 0.17, 0.54 +/- 0.29, and 0.61 +/- 0.22. A total of 40% of the Klebsiella strains, 80% of the Serratia strains, and 46% of the E. coli strains had a fractional inhibitory concentration equal to or less than 0.25 for both of these antimicrobial agents and were considered to be synergistically inhibited by the combination. By applying this criterion to bactericidal activity, synergy was demonstrated against 50, 65, and 46% of these strains, respectively. All of the Enterobacteriaceae tested were inhibited by clinically achievable concentrations of trimethoprim and amikacin. Antagonism was not demonstrated with any of the organisms tested. Trimethoprim had no antibacterial effect on the Pseudomonas strains and did not alter amikacin's activity against these bacteria.     

Amikacin-induced alterations in the structure of gram-negative bacilli

Exposure of some species of Enterobacteriaceae and of Pseudomonas aeruginosa to amikacin results in structural alterations which coincide with a decrease in the number of colony forming units. Escherichia coli when exposed to amikacin at a concentration twice the minimum inhibitory concentration for 2 hr shows a reduction in the number of ribosomes in the center of the cell and an aggregation of nuclear material in a peculiar concentric pattern we called "tornado image." After 2-4 hr of exposure to amikacin at concentrations of two to five times the minimal inhibitory concentration, all species tested showed in addition to alterations in the distribution of ribosomes, ruptures in the cytoplasmic membrane, damaged cell walls, and in some instances complete loss of cellular shape. It appears that amikacin produces a lytic death of bacteria.     

2011年临床常见病原菌分布特点及耐药性分析

目的回顾性分析临床分离常见病原菌分布及对常用抗生素耐药情况,为临床合理用药提供依据。方法对2011年1～12月临床送检各类标本进行分离培养,采用全自动微生物仪进行菌株鉴定,抗菌药物的敏感性检测采用K-B纸片扩散法,判定标准依据CLSI2009指南判定。结果共分离出病原菌2 790株,以革兰阴性杆菌为主,前四位依次为克雷伯菌属、大肠埃希菌、铜绿假单胞菌和不动杆菌。肠杆菌科细菌对氨苄西林耐药率最高,均大于76.3%,对阿米卡星、哌拉西林/他唑巴坦、头孢哌酮/舒巴坦保持较高的敏感性,耐药率小于25.9%,大肠埃希菌产超广谱β-内酰胺酶(ESBLs)检出率为52.3%;克雷伯菌属产ESBLS检出率为33.9%。ESBLs的大肠埃希菌和克雷伯菌属对抗菌药物的耐药率明显高于非产ESBLS的菌株,未检出耐亚胺培南、美罗培南菌株。非发酵菌中,阿米卡星、头孢哌酮/舒巴坦、美洛培南对铜绿假单胞菌具有良好的抗菌活性;亚胺培南、美洛培南、米诺环素对不动杆菌属敏感性强。耐甲氧西林金黄色葡萄球菌(MRSA)的检出率为44.1%,耐甲氧西林凝固酶阴性葡萄球菌(MRCNS)的检出率为69.8%,耐甲氧西林的葡萄球菌对所有β-内酰胺类抗生素均耐药,未检出耐万古霉素和利奈唑胺的葡萄球菌。结论及时准确地对本地区病原菌进行耐药性监测,为临床合理应用抗生素提供有力的依据,预防和延缓耐药菌的产生和传播。     

Synergistic effects of chlorpromazine and perphenazine on several chemotherapeutic agents. I. General profile of the effects measured by the filter paper strip-agar diffusion method with Escherichia coli and Pseudomonas aeruginosa.

The combination effects of chlorpromazine (CPZ) and periphenazine (PPZ) with beta-lactam antibiotics (ampicillin, carbenicillin, cefazolin) and nalidixic acid group compounds (nalidixic acid, piromidic acid and pipemidic acid) have been estimated to be synergistic by the filter paper strip-agar diffusion method (Dye's method) with Escherichia coli and Pseudomonas aeruginosa as test organisms. The observed synergism might be associated with their inhibition of various enzymes including ATPase and DNAase as well as with their specific binding to DNA. Similar synergistic effects of CPZ and PPZ have been shown by the broth dilution method. Based on these findings, it seems to be a fascinating project to devise a new phenothiazine drug without influence in mental disease that will have a greater measure of synergistic effect when combined with the above-studied antibacterial agents.     

In vitro antimicrobial activity of aztreonam alone and in combination against bacterial isolates from pediatric patients

We examined 134 pediatric clinical isolates of Enterobacteriaceae, Pseudomonas aeruginosa, and gram-positive cocci for susceptibility to aztreonam alone and in combination with seven other antibiotics. All 98 gram-negative isolates were susceptible to aztreonam with similar inhibitory and bactericidal activity. Combinations of aztreonam with cefoxitin, ampicillin, or clindamycin were generally indifferent or additive. Synergism was occasionally seen against enteric organisms with aztreonam plus cefoxitin or clindamycin. Combinations of tobramycin and aztreonam were synergistic (62%) against P. aeruginosa; aztreonam plus piperacillin or ticarcillin was additive. Aztreonam did not affect the activity of nafcillin against Staphylococcus aureus, or of ampicillin against species of Streptococcus group B or D. Antagonism was seen only with aztreonam plus cefoxitin against Enterobacter species, but not at clinically significant concentrations. Several combinations of antibiotics with aztreonam should be appropriate for initial therapy of infections in children without major risks of antibacterial antagonism.     

Amikacin synergism with beta-lactam antibiotics against selected nosocomial pathogens

The synergistic interaction between amikacin and several investigational antibiotics against seven different genera of nosocomial pathogens was assessed using the microtitre checkerboard technique. The greatest percentage of tests showing synergy was when amikacin was used in combination with apalcillin and azlocillin particularly against Serratia marcescens and Proteus spp. When amikacin was combined with several new semi-synthetic cephalosporins, synergy was present in a variable percentage of tests. No antagonism was found in any of the tested combinations.     

In vitro activity of newer β-lactam agents in combination with amikacin against Pseudomonas aeruginosa, Klebsiella pneumoniae, and Serratia marcescens

The in vitro activity of cefoperazone, ceftazidime, ceftizoxime, moxalactam, and N-forminidoyl thienamycin was evaluated alone and in combination with amikacin to assess possible synergistic activity against isolates of amikacin-resistant and multidrug-resistant and susceptible to amikacin (one isolate was also resistant to amikacin). The checkerboard agar dilution method was used. Ceftazidime and thienamycin followed by moxalactam and cefoperazone were the most active agents versus the alone and in combination testing. Ceftazidime, moxalactam, and thienamycin showed the greatest activity against and all agents except cefoperazone were active against . The finding of synergy or partial synergy in combination testing was found in the majority with all three genera, including levels below the breakpoint for both amikacin and the β-lactam agents. This wide in vitro activity indicates that clinical evaluation of these agents in treatment of multidrug-resistant infections is warranted.     

Intracellular activity of cefamandole and aztreonam against phagocytosed Escherichia coli and Staphylococcus aureus

The intracellular activity of cefamandole and aztreonam against phagocytosed Escherichia coli and cefamandole against phagocytosed Staphylococcus aureus was studied using a sensitive and standardized method of murine peritoneal macrophages. Cefamandole and aztreonam exerted an intracellular antibacterial activity against E. coli which was greater than their extracellular one. With concentrations of both antibiotics up to 16 x MBC a dose-dependent decrease of the initial number of intracellular E. coli which ranged from 32% to 90% was observed. However, similar antibiotic concentrations above the MBC affected the viability of extracellular E. coli by only 20% to 30%. The intracellular antibacterial activity of both antibiotics against E. coli was further enhanced by immune serum. Cefamandole at 4 x the MBC did not affect the survival of intracellular S. aureus, but killed 41% of extracellular bacteria by 1 h and 99% after 3 h. The intracellular activity of both antibiotics against E. coli was also maintained in NaF-pulsed macrophages which have an impaired oxidative metabolism. The data suggest that both cefamandole and aztreonam possess an intracellular antibacterial activity against E. coli that seems at least in part due to a positive cooperation of antibiotics with the O2-independent microbicidal system of macrophages.     

In vitro activity of newer beta-lactam agents in combination with amikacin against Pseudomonas aeruginosa, Klebsiella pneumoniae, and Serratia marcescens

The in vitro activity of cefoperazone, ceftazidime, ceftizoxime, moxalactam, and N-formimidoyl thienamycin was evaluated alone and in combination with amikacin to assess possible synergistic activity against isolates of amikacin-resistant Pseudomonas aeruginosa and multidrug-resistant Serratia marcescens and Klebsiella pneumoniae susceptible to amikacin (one S. marcescens isolate was also resistant to amikacin). The checkerboard agar dilution method was used. Ceftazidime and thienamycin followed by moxalactam and cefoperazone were the most active agents versus the P. aeruginosa alone and in combination testing. Ceftazidime, moxalactam, and thienamycin showed the greatest activity against S. marcescens, and all agents except cefoperazone were active against K. pneumoniae. The finding of synergy or partial synergy in combination testing was found in the majority with all three genera, including levels below the breakpoint for both amikacin and the beta-lactam agents. This wide in vitro activity indicates that clinical evaluation of these agents in treatment of multidrug-resistant infections is warranted.     

Selective microbiologic effects of tea extract on certain antibiotics against Escherichia coli in vitro

OBJECTIVES: This study evaluated the microbiologic effects of black tea, compared to green tea, alone and in conjunction with selected antibiotics against Escherichia coli, the common cause of intestinal and urinary tract infections. DESIGN: This study was an in vitro evaluation of antibacterial effects of tea extracts. METHODS: Black and green tea extracts were analyzed by using high-performance liquid chromatography to compare their major polyphenol profiles. Different concentrations of the extracts or gallic acid (GA), the phenolic compound found with high concentration in the black tea extract, were employed for bacterial sensitivity tests, using pour plate and disc diffusion methods. The latter was used to evaluate the interactions between the extracts and certain anti-E. coli antibiotics. RESULTS: GA in black tea extract and epigallocatechin and epigallocatechin gallate in green tea extract are present in the highest concentrations, respectively. At concentrations of 25 mg/mL, both black and green teas after 5 and 7 hours completely inhibited E. coli growth. GA at concentrations of 5, 10, and 25 microg/mL after 7, 5 and 3 hrs, respectively, inhibited bacterial growth. Both black and green tea extracts had either synergistic or antagonistic effects at different concentrations on selected antibiotics, while GA showed a synergistic effect with all the antibiotics tested in a dose-dependent manner. The effect was more prominent with amikacin and sulfamethoxazole. CONCLUSIONS: The microbiologic effects of both black tea and green tea extracts on certain antibiotics against E. coli may vary, depending on the type of the tea extract (i.e., black vs. green), the amount of the extract, and the antibiotic being used.     

Comparative in vitro susceptibilities of 504 bacteremic isolates to ticarcillin plus clavulanic acid and other antimicrobial agents

A total of 504 clinical bacteremic isolates were tested for susceptibility to ticarcillin-clavulanic acid and 12 other antibiotics. Ticarcillin-clavulanic acid showed superior antibacterial activity compared to penicillin, mezlocillin, piperacillin, ticarcillin, gentamicin, and amikacin against bacteremic isolates of methicillin-susceptible Staphylococcus aureus and Staphylococcus epidermidis. However, ticarcillin-clavulanic acid's activity was inferior to that of vancomycin against methicillin-resistant isolates of S. aureus and S. epidermidis. For Escherichia coli, Klebsiella oxytoca, Proteus mirabilis, Providencia stuartii, and lactose nonfermenting aerobic gram-negative bacilli, the activity of ticarcillin-clavulanic acid surpassed that of mezlocillin, piperacillin, and ticarcillin. Of the antimicrobial agents tested, ticarcillin, piperacillin, ceftazidime, and amikacin were the most active antibiotics against Pseudomonas aeruginosa.     

Effect of Two Cancer Chemotherapeutic Agents on the Antibacterial Activity of Three Antimicrobial Agents

Cancer chemotherapeutic agents and antibacterial antibiotics are often given concomitantly. Daunorubicin, cytosine arabinoside, and three antibiotics (gentamicin, amikacin, and ticarcillin) were tested individually and in combinations to determine their antimicrobial activity against Pseudomonas aeruginosa, Klebsiella pneumoniae, and Escherichia coli. These cytotoxic agents are commonly employed in the therapy of acute nonlymphocytic leukemia for remission induction therapy, and these antimicrobial agents are used in infection therapy. The maximum concentrations of the two cytotoxic drugs were chosen to be twice the known peak plasma levels of commonly employed dosage schedules. Neither of the cancer chemotherapeutic agents, alone or in combination, demonstrated bactericidal activity at the levels tested. However, in the presence of these agents, the antimicrobial activity of gentamicin and amikacin, although not that of ticarcillin, was depressed for 11 of 15 K. pneumoniae strains and 8 of 15 P. aeruginosa strains, but for none of the strains of E. coli. This level of decreased activity occasionally resulted in a minimal inhibitory concentration of the tested aminoglycoside well above the standard serum levels. Daunorubicin was more likely to antagonize gentamicin than was cytosine arabinoside.