The patterns and transmissibility of antibiotic resistance among clinical strains of Pseudomonas aeruginosa.

Four hundred and ninety-eight predominantly pyocin-type 10 clinical strains of Pseudomonas aeruginosa were analyzed for resistance to carbenicillin, cefoperazone, cefotaxime, ceftazidime, gentamicin, amikacin and netilmicin. Based on NCCLS-recommended MIC breakpoints, 245 strains were found to be resistant, of which 41.6% were resistant to carbenicillin, 38% to gentamicin, 37.8% to netilmicin, 26.3% to cefoperazone, 17.9% to cefotaxime, 0.6% to amikacin and none to ceftazidime. Quadruple resistance to carbenicillin, cefoperazone, gentamicin and netilmicin was the most frequent pattern observed. Resistance to older antibiotics (kanamycin, streptomycin and tetracycline) and to mercuric chloride were also common. Conjugation experiments suggested that self-transmissible and non-transmissible plasmids occurred in at least 66 strains.     

Comparative Activity of N-Formimidoyl Thienamycin with Third Generation Cephalosporins and Ureido Penicillins against Multiple Resistant Serratia marcescens

Twenty multiple resistant clinical isolates were tested with N-formimidoyl thienamycin, moxalactam, cefotaxime, cefoperazone, and the three ureidopenicillins: azlocillin, mezlocillin, and piperacillin. A concentration of < 0.97 μg/ml inhibited 100% of organisms for N-f-thienamycin and cefotaxime, 90% for moxalactam, and 60% for cefoperazone. An increase in inoculum from 10³ to 10⁶ cells reduced activity fourfold for 95% of isolates with cefoperazone, 70% with N-formimidoyl thienamycin, 65% for cefotaxime, but only 15% for moxalactam. For ureidopenicillins, 85% of strains tested had MIC's ≤ 15.6 μg/ml. An inoculum effect was observed in only 35–50%. At 10³, the cidal concentration was the same or twofold greater than the inhibitory level for N-f-thienamycin and cephalosporins in 70% of strains tested and 65% for penicillins. With 10⁶, the 70% value remained for N-f-thienamycin but was reduced to 45% for cefotaxime and 25% for moxalactam; 85% demonstrated > eightfold differences with cefoperazone. Single step high-level resistance was observed to moxalactam (20%). Carbenicillin resistant strains were cross-resistant to the ureidopenicillins. N-f-thienamycin and cefotaxime appeared comparable, although important differences between morphological alteration and metabolism may influence their therapeutic effectiveness.     

我院2008-2011年抗菌药物使用量与铜绿假单胞菌耐药相关性的研究

目的:探讨2008-2011年我院抗菌药物使用与铜绿假单胞菌(PA)耐药水平变化之间的关系.方法:计算10种抗菌药物平均每日每百张床住所消耗的限定日剂量(DDD)及同期PA的耐药率,并对抗菌药物用量与耐药率进行相关性分析.结果:发现头孢哌酮/舒巴坦及左氧氟沙星DDD分别与铜绿假单胞菌对头孢噻肟、头孢吡肟、左氧氟沙星、亚胺培南、头孢噻肟、头孢哌酮/舒巴坦、头孢吡肟耐药率的相关性有统计学意义,其它组数据无统计学意义.结论:呼吸病区左氧氟沙星及头孢哌酮/舒巴坦DDD的使用量与PA耐药率相关.     

Pseudomonas aeruginosa and beta-lactam resistance phenotypes (Tunisia 1989-1990)]

366 strains of P. aeruginosa were studied. More than half of samples were issued from urology (40%) and general surgery (21%) and samples are mostly urine specimens (46%). The phenotypes of resistance to 3 beta-lactams (respectively ticarcillin, cefotaxime and ceftazidime) showed that 73% of the strains were wild phenotype (SSS-SIS-SRS). The MIC of ticarcillin +/- clavulanic acid, azlocillin, cefoperazone, cefsulodin, cefotaxime and ceftazidime against 93 strains chosen among different phenotypes of resistance showed that ceftazidime is the most active beta-lactam, 93% of strains being inhibited by 16 micrograms/ml. These MIC and iodometric test performed on 33 strains with ticarcillin MIC > 128 mg/l showed that acquired resistance was non enzymatic in 21% of cases and was related to beta-lactamase production in 79% of cases.     

Sensitivity of bacteria of the genus Acinetobacter to antibiotics and ofloxacin

Between 1989-1989 276 strains of Acinetobacter genus were isolated which contained: Acinetobacter calcoaceticus subsp. anitratus (n = 167), Acinetobacter calcoaceticus subsp. Iwoffi (n- = 83), Acinetobacter haemolyticus (n = 26). Their sensitivity to aminoglycoside antibiotics, beta-lactams, tetracyclines, chloramphenicol, colistin, and ofloxacin was tested. More than 90% of strains were sensitive to colistin and ofloxacin. The sensitivity to remaining antibiotics differentiated depending on species. Acinetobacter anitratus were highly resistant to Ist and IInd generation of cephalosporins, and moreover to penicillins, tetracyclines, and chloramphenicol. Cephalosporins of IIIrd generation were active against 70% of strains with exception of cefoperazone what was also the case for representatives of aminoglycosides as netilmicin and amikacin. Strains of Acinetobacter Iwoffi were in majority sensitive to all antibiotics with exception of cephalothin, cephradine and cefoperazone. More than 90% of Acinetobacter haemolyticus strains were sensitive to gentamicin, carbenicillin, azlocillin, ceftriaxone, cefotaxime and tetracyclines.     

Comparative in vitro activity of first, second and third generation cephalosporins

Minimum inhibitory concentrations (MIC) were determined against 662 recent clinical isolates for eight cephalosporins representing first, second and third generation compounds. All four third-generation cephalosporins tested (cefoperaxone, cefotaxime, ceftazidime and moxalactam) were significantly more active against aerobic gram-negative bacteria than the older compounds (cephalothin, cefamandole, cefoxitin, and cefuroxime). Cefotaxime and moxalactam were most active against Enterobacteriaceae with extremely low MIC-values. Ceftazidime was definitely most active against Pseudomonas aeruginosa with more than 90% of strains inhibited at 4 micro g/ml. MIC-values for cefotaxime against Staphylococcus aureus were for all strains 1-2 micro g/ml, slightly higher for cefoperazone, while the effect of ceftazidime and moxalactam was more limited. All third generation cephalosporins demonstrated efficiency against Streptococcus pyogenes, cefotaxime being most active and moxalactam least active, but were essentially ineffective against Streptococcus faecalis. Moxalactam demonstrated higher activity against Bacteroides fragilis than other second and third generation cephalosporins including cefoxitin. Previous studies have demonstrated a very high activity of all third generation cephalosporins against Haemophilus influenzae and Neisseria gonorrhoeae, including beta-lactamase producing strains.     

Inactivation of some third-generation cephalosporins by β-lactamases of non-fermentative Gram-negative bacteria isolated from freshwater of a remote environment

Inactivation of cefotaxime, cefoperazone and ceftazìdime by β-lactamases from strains of Pseudomonas sp. and Acinetobacter calcoaceticus isolated from freshwater of a remote environment is demonstrated. Inactivation rates showed some relation-ship to susceptibility of the strains as a whole, but this relationship was absent for individual strains, probably because of some other resistance mechanisms. Inac-tivation was also observed with low inocula if incubation was prolonged. These β-lactamases seem to be coded by chromosomal genes, since no plasmids were found and they exhibited relatively high isoelectric points.     

耐亚胺培南铜绿假单胞菌对左氧氟沙星的耐药性观察

目的对比研究左氧氟沙星和常用的12种抗生素对92株耐亚胺培南铜绿假单胞菌的耐药性.方法药敏试验采用K-B纸片扩散法.结果左氧氟沙星对耐亚胺培南菌株的耐药率为43.48%;显著低于头胞噻肟、头胞西丁、头胞他定、头胞哌酮、氧氟沙星和洛美沙星(P＜0.02或P＜0.001);和诺氟沙星、环丙沙星、阿米卡星、舒普深、妥布霉素的耐药率无差异([ WTBX P＞0.05).结论左氧氟沙星对耐亚胺培南铜绿假单胞菌有良好的抗菌活性.     

2005～2008年儿童菌痢病原菌与药敏分析及临床意义

目的了解儿童细菌性痢疾病原菌的分布特征及药敏特点,为临床更严谨更规范使用抗生素提供支持与依据。方法对2005年10月至2008年10月57例儿童菌痢的菌型、药敏及耐药性进行分析。结果儿童细菌性痢疾病原菌亚型分类中宋氏痢疾杆菌（D群）占14．0％,福氏痢疾杆菌（B群）占86．0％;痢疾杆菌对常用抗生素耐药率由低到高依次为头孢噻肟,丁胺卡那霉素,庆大霉素,头孢哌酮,头孢三嗪,头孢他啶,头孢唑啉,环丙沙星,氯霉素,复方新诺明,氨苄青霉素;痢疾杆菌单株对多种抗生素的总耐药率为39．2％,多重耐药率为43,9％,且各组间差异无显著性（χ^2=1．608,P=0．996）,痢疾杆菌的耐药问题依然严重。结论新乡市区儿童细菌性痢疾病原菌亚型分类D组已呈明显上升趋势,但总体仍以B群感染为主（86％）;痢疾杆菌的耐药问题依然严重;对儿童菌痢选用抗生素应结合药敏首选头孢噻肟等第三代头孢类抗生素或头孢唑啉,年长儿也可选用丁胺卡那霉素、庆大霉素等氨基苷类,而以环丙沙星为代表的喹诺酮类药物不宜作为儿童尤其7岁以下小儿菌痢的备选药物;氯霉素、复方新诺明及氨苄青霉素已不再作为小儿菌痢的抗菌选择。     

Determination of cephalosporins in biological material by reversed-phase liquid column chromatography

Seven cephalosporins (β-lactam antibiotics), viz. cefazolin, cephalotin, cefoxitin, cefotaxime, cefamandole, cefuroxime and cefoperazone (T 1551) were determined in biological material. The compounds were extracted from acid-treated body fluids into chloroform—1-pentanol (3:1) and re-extracted into a small volume of an aqueous phase at pH 7, which was injected into the chromatographic column. The chromatographic support was μBondapak C_1a (10 μm) and the mobile phase was a mixture of 0.01 M acetate buffer (pH 4.8) and methanol or acetonitrile. Detection limits are about 50 ng/ml for extractions from 1 ml of serum and have permitted pharmacokinetic studies of the seven cephalosporins.     

Molecular mechanisms of cephalosporin resistance in Gram-negative bacteria of the Enterobacteriaceae family]

Extended spectrum beta lactamase genes were detected by the PCR in 87.6% of 231 Enterobacteriaceae strains isolated in medical institutions of Moscow, St. Petersburg, Tomsk and Nazran that showed a decrease in their susceptibility to 3rd generation cephalosporins. Alone or in various combinations TEM type beta lactamases were detected in 43.3% of the isolates, 46.8 and 51.2% of the isolates produced SHV type and CTX type beta lactamases respectively. Combinations of 2 and 3 different determinants were detected in 40 and 14% of the isolates respectively. Production of class C beta lactamases was suspected in 28% of the isolates by their resistance to cefoxitin. The gene of ACT type beta lactamase was detected in 1 strain of Klebsiella pneumoniae and the gene of CMY type beta lactamase was detected in 1 strain of Proteus mirabilis. By the NCCLS 100% of the isolates was susceptible to meropenem, 14% was susceptible to cefotaxime, 64% was susceptible to cefepime, 81% was susceptible to cefoperazone/sulbactam, 47% was susceptible to gentamicin, 57% was susceptible to amikacin and 36% was susceptible to ciprofloxacin.     

2002年至2007年太原地区儿童细菌性腹泻病原菌分布及耐药分析

目的了解太原地区近6年儿童细菌性腹泻病原菌分布及耐药情况。方法对临床诊断细菌性腹泻病,便培养已分离到病原菌1080例作回顾性分析,分析其病原菌的分布及耐药情况。结果埃希菌属486株（45％）,居于首位,前5位的病原菌依次为埃希菌属、肠球菌属、酵母样真菌、志贺菌属、假单胞菌属。各年均以大肠埃希菌为主要检出菌,志贺菌逐年减少。年龄分布中,婴儿的构成比最高（44．4％）。埃希菌属、志贺菌属、假单胞菌属、沙门菌属、气单胞菌属此5种杆菌对13种抗生素的平均耐药率依次为舒普深、痢特灵、头孢他啶、庆大霉素、环丙沙星、头孢哌酮、头孢曲松、丁胺卡那、头孢噻肟、诺氟沙星、头孢呋辛、哌拉西林、头孢唑啉。从埃希菌属近6年的耐药性变迁资料可以看出,对13种抗生素的耐药率均有不同程度上升。结论传统的致病菌志贺菌属、沙门菌属较少,而肠球菌属、假单胞菌属、枸橼酸杆菌属、克雷伯杆菌属、肠杆菌属、酵母样真菌等条件致病菌肠炎占有相当比例。各种致病菌的耐药性增加,第三代头孢除头孢他啶的耐药率较低外,其余都较高。提示应严格掌握抗生素用药指证,合理选用抗生素。     

Sensitivity of Pseudomonas to currently used antibacterial drugs

Sensitivity of 184 strains of four Preudomonas species that are of importance from the medical viewpoint was studied. It was shown that all the studied strains were resistant to cefazolin and ampicillin. Sulfomonomethoxine was active only against P. mallei. Gentamicin inhibited the growth of all the strains of P. mallei and 60 per cent of the strains of P. aeruginosa. Among the four studied strains, the causative agent of glanders was the least resistant to the chemotherapeutic drugs. Eleven out of the 16 studied drugs were active against P. mallei. The strains of P. pseudomallei, the organism causing melioidosis were sensitive to 6 drugs: ceftazidime, imipenem, doxycycline, minocycline, chloramphenicol and biseptol. The dime, ceftriaxone, cefsulodin, cefoperazone, cefotaxime, piperacillin and imipenem. The strains of P. cepacia were the most resistant to all the chemotherapeutic drugs studied and only biseptol was active in the concentrations attained in blood of patients (MIC50 12.5 mg/l). 10 to 70 per cent of the strains were sensitive to ceftazidime, imipenem, doxycycline, minocycline and rifampicin. Therefore, imipenem and ceftazidime were the most active drugs with respect to the causative agents of pseudomonoses. Biseptol, doxycycline and minocycline are also worthy note.     

Biliary secretion of cefoperazone and its inter-relationship with bile acid transport in the rat

Cefoperazone is a third generation cephalosporin which is secreted predominantly in bile. This study set out to examine the effect of stimulating bile choleresis on the biliary secretion of cefoperazone. Stimulation of both bile acid-dependent and independent bile flow (phenobarbitone pretreatment) hastened the peak appearance of a pulse of cefoperazone into bile. Although the biliary secretion rate of cefoperazone was enhanced by bile acid infusion, the % recovery and maximal biliary concentration were reduced. The reciprocal effect of continuous cefoperazone infusion on the rate of biliary transport of a pulse of bile acid was examined. Cefoperazone infusion hastened the biliary transport of glycocholate. Net recovery of glycocholate was unaffected.     

Purification and characterization of a beta-lactamase from Haemophilus ducreyi in Escherichia coli

A pCb plasmid encoding a beta-lactamase from Haemophilus ducreyi was transferred to Escherichia coli, purified, and characterized. The beta-lactamase could be isolated from a culture filtrate and further purified by ammonium sulfate and chelating Sepharose fast flow loaded with Zn(2+). The purified enzyme resulted in a major band at approximately 30-kDa on SDS-PAGE and its pI was determined to be 5.4. The beta-lactamase could hydrolyze both penicillin antibiotics including ampicillin, benzylpenicillin, and carbenicillin as well as cephalosporin antibiotics including nitrocefin, cephalothin, cephaloridine, and cefoperazone. However, benzylpenicillin was the best substrate. The enzyme activity was inhibited by clavulanic acid but not by boric acid, cefotaxime, ethylenediaminetetraacetic acid, or phenylmethylsulfonyl fluoride. The sequence of the beta-lactamase gene was also determined. It confirmed that the enzyme belonged to a class A beta-lactamase which had 99% identity to the ampicillin resistance transposon Tn3 of pBR322. Two nucleotides were different between the E. coli (Tn3) and H. ducreyi (pCb) genes that affected the amino-acid sequence. The valine at position 82 (ABL 84) was changed to isoleucine and the alanine at position 182 (ABL 184) was changed to valine. Genetic homogeneity among beta-lactamases is remarkable. Amino acid sequencing of some beta-lactamases has shown that substitution of only a few amino acids in the bla gene leads to high-level resistance against specific cephalosporins.     

Comparative clinical and epidemiological evaluation of beta-lactam antibiotics in the treatment of intraabdominal infections]

We performed a retrospective, comparative study to evaluate efficacy, safety and economic outcomes of empiric cefoperazone/sulbactam monotherapy compared with the meropenem, imipenem/cilastatine and combination of cefepime plus metroindazol in patients with intra-abdominal infection. A total of 468 patients diagnosed with intra-abdominal abscess, peritonitis, pancreatitis were included in the study (the severity of infection according to scale APACHE II was less than 15). Patients were randomized to be treated with either 500 mg meropemen i.v. every 8 hours or 500 mg imipenem/cilastatine i.v. every 8 hours or 2 g cefepime i.v. every 12 hours plus 500 mg metronidazol twice daily or cefoperazone/sulbactam 2 g daily administered every 12 hours. Overall positive clinical responses (cure or improvement) were achieved at the end of treatment for 87.5 patients in meropenem group, 86.6% in the imipenem/cilastatin group, 85.3% in the cefepime group and 86.8% in cefoperazone/sulbactam group. Total cost of the treatment per 100 patients with intra-abdominal infections for cefoperazone/sulbactam was 1957031 roubles, for combinations of cefepime with metronidazol--2497815 roubles. For carbapenem group cost achieved for meropenem--3085291 rub., for imipenem/cilastatin--2653388 roubles. Rate "cost-effectiveness" in total: 784.47$ for cefepime, and 834.39$ for imipenem/cilastatine, 970.21$ for meropenem and 615.4$ for cefoperazone/sulbactam. The most expensive treatment was considered to be with meropenem and imipenem/cilastatine, main share is determined by initial cost of preparations. Less expensive was treatment by cefoperazone/sulbactam with cefepime and by metronidazol.     

Liquid chromatography/tandem mass spectrometry assay for the simultaneous determination of cefoperazone and sulbactam in plasma and its application to a pharmacokinetic study

A rapid and highly sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for simultaneous determination of cefoperazone sodium and sulbactam sodium in human plasma was developed. The analytes and internal standard (IS), cefuroxime sodium, were extracted from human plasma via liquid-liquid extraction with ethyl acetate and separated on a Waters Xterra C18 column within 3.5 min. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique, operating in selected reaction monitoring (SRM) and negative ion mode. The precursor to product ion transitions monitored for cefoperazone, sulbactam and IS were m/z 644.1→528.0, 232.1→140.0, and 423.0→362.0, respectively. The assay was validated in the linear range of 0.1-20 μg/mL for cefoperazone and 0.02-4 μg/mL for sulbactam. The intra- and inter-day precisions (CV%) were within 8.39% for each analyte. The recoveries were greater than 87.3% for cefoperazone and 87.2% for sulbactam. Each analyte was found to be stable during all sample storage, preparation and analytical procedures. The method was successfully applied in a pharmacokinetic study of Sulperazon injection in six hospital-acquired pneumonia (HAP) patients.     

Effect of sulbactam on anti-pseudomonal activity of beta-lactam antibiotics in cells producing various levels of the MexAB-OprM efflux pump and beta-lactamase

The beta-lactamase inhibitor, sulbactam, was tested for beta-lactamase inhibitory activity in Pseudomonas aeruginosa cells producing various levels of both the MexAB-OprM efflux pump and beta-lactamase. We found that sulbactam lowered the MICs of cefoperazone and piperacillin by inhibiting the beta-lactamase 8-fold in the cell producing a constitutively high level of AmpC-type beta-lactamase and a wild-type level of MexAB-OprM pump compared with that without sulbactam. The MICs of cefoperazone and piperacillin in the cell producing a constitutively high level of both the efflux pump and beta-lactamase under the presence of sulbactam were 8 and 4 times, respectively, lower than that without sulbactam. The MICs of sulbactam in the cell producing a constitutively high and a wild-type level of the efflux pump were 16- and 8-fold higher, respectively, than that in the mutant lacking the efflux pump. We concluded that sulbactam exerts potent beta-lactamase inhibitory activity in the cell producing a high level of efflux pump, in spite of the fact that sulbactam serves as a substrate of the MexAB-OprM pump. Increasing amounts of sulbactam over the weight of beta-lactams further strengthen the effect of beta-lactam antibiotics.     

Effect of beta-lactam antibiotics on lipid bilayer membranes. II. Cephalosporin antibiotics

Interaction of cephalosporin antibiotics with flat bilayer lipid membranes (BLM) composed of vegetative or bacterial phospholipids was studied with respect to their effect on electroconductivity, capacity for binding to the bilayers and capacity for transmembrane transfer through the bilayers. By their effect on conductivity, the investigated cephalosporins could be divided into three groups: those having no effect on conductivity at the maximum concentrations (up to 1 mg/ml) i.e. cefoperazone, cefotaxime and ceftizoxime, those having an effect on conductivity at concentrations of 0.2 to 0.5 mg/ml i.e. cephalexin and cephalothin and those having a significant effect on conductivity at concentrations of 10 to 60 micrograms/ml i.e. N-acyl derivative of 7-ACA (substance 64) and 7-ACA derivative (substance 71). The cephalosporins had a capacity for binding to the bilayers and for incorporation into the bilayers. They penetrated to some extent through the bilayers composed of either vegetable or bacterial phospholipids. The lower rate of penetration through the BLM as compared to that of penicillins must be due to lower hydrophobicity of the cephalosporin molecules as compared to that of penicillins.     

Prevention and treatment with cefoperazone of postoperative suppurative complications in heart surgery]

Clinical trials of cefoperazone (cefobid, Pfizer, USA) were carried out in 49 patients with cardiovascular diseases who had undergone surgical operations. The pathogens of infectious complications were investigated bacteriologically. Good results of the treatment were observed in 43 patients. Allergic reaction developed in 1 patient. Cefoperazone was shown advantageous in treatment of pulmonary complications in the operated patients. It was found possible to use cefoperazone in combination with aminoglycosides. Cefoperazone was found to be one of the drugs of choice in the treatment of aerobic and anaerobic bacteriemia, as well as sepsis after surgical operations on the heart and great vessels. The results on the use of cefoperazone for short-term "perioperative" prophylaxis in cardiosurgery (in accordance with the WHO instructions) are also presented.