Risk-factors for the acquisition of imipenem-resistant Acinetobacter baumannii in Spain: a nationwide study

Potential risk-factors for the acquisition of imipenem-resistant Acinetobacter baumannii were investigated in a cohort study in 25 Spanish hospitals. The clonal relationship among isolates was determined by pulsed-field gel electrophoresis (PFGE). In total, A. baumannii was isolated from 203 patients, with imipenem-resistant (MIC(90) 128 mg/L) isolates being obtained from 88 patients (43%), and imipenem-susceptible isolates from 115 patients (57%). A wide clonal distribution was observed among the imipenem-resistant isolates, but spread of the same clone among centres was not demonstrated. The results indicated that imipenem-resistant A. baumannii is a widely distributed nosocomial pathogen in Spain and reaches an alarming frequency in some centres. Independent risk-factors for the acquisition of imipenem-resistant A. baumannii were a hospital size of >500 beds (multivariate OR, 6.5; 95% CI, 1.8--23), previous antimicrobial treatment (multivariate OR, 4.3; 95% CI, 1.6--11), a urinary catheter (multivariate OR, 2.7; 95% CI, 1.1--6.7) and surgery (multivariate OR, 2; 95% CI, 1.07--3.8).     

多耐药鲍氏不动杆菌对替加环素的耐药分析

目的探讨替加环素对多耐药鲍氏不动杆菌的体外抗菌活性。方法从2009年4月至2010年3月中山大学附属第三医院住院患者中分离41株多耐药鲍氏不动杆菌（multidrug-resistant Acineto bacterbaumannii,MDR-AB）,采用SIEMENS公司MicroScan Walk Away40全自动微生物鉴定仪鉴定菌株,用K-B法测定鲍氏不动杆菌对12种抗菌药物的敏感性。结果鲍氏不动杆菌主要分离自痰标本,占80.5%;病区主要分布在ICU（37%）和神经外科（22%）。除亚胺培南外,其余11种抗菌药物的非敏感率均大于80%。替加环素的非敏感率为80.4%,敏感率为19.5%;亚胺培南的敏感率最高,为36.6%,非敏感率为63.4%。结论多耐药鲍氏不动杆菌对替加环素的非敏感率很高,可能与外排泵系统有关。替加环素对MDR-ABA感染的治疗效果不容乐观。     

2012年深圳市光明新区人民医院细菌耐药监测

目的了解2012年深圳市光明新区人民医院临床分离菌对常用抗菌药物的耐药性。方法采用Microscanauto4鉴定及药敏系统对临床常规细菌进行监测．按CLSl2009年版标准判断药敏结果．并用WHONET5．4软件统计分析。结果全年共分离细菌1818株,其中革兰阳性菌占35．6％,革兰阴性菌占58．O％。葡萄球菌中耐甲氧西林金黄色葡萄球菌（MRSA）占18．3％,凝固酶阴性葡萄球菌（MRCNS）占71．9％。肺炎链球菌中青霉素非敏感的肺炎链球菌（PNSSP）占20．7％。大肠埃希菌和肺炎克雷伯杆菌产ESBLs菌株分别占43．2％和22．4％。检出5株亚胺培南耐药的肠杆科细菌。铜绿假单胞菌和鲍曼不动杆菌对头孢他啶、哌拉西林、妥布霉素、阿米卡星、环丙沙星、亚胺培南的耐药率均低于10％。结论本院常见致病菌耐药性不是十分严重,尤其是院内感染的铜绿假单胞菌和鲍曼不动杆菌对常用抗菌药物耐药率较低。     

Comparison of trends of resistance rates over 3 years calculated from results for all isolates and for the first isolate of a given species from a patient

We compared trends of annual resistance rates calculated from results for all isolates and for the first isolate of Staphylococcus aureus, Klebsiella pneumoniae, and Acinetobacter baumannii per patient over a 3-year period from 2001 through 2003. Antimicrobial susceptibility results of inpatients were extracted from a computerized database. Annual resistance rates of a species were calculated by two methods: (i) from results for all isolates, even those from patients with multiple isolates in a given year and (ii) from results for the first isolate from a patient in a given year, regardless of susceptibility profile or specimen type. Rates of methicillin-resistant S. aureus (MRSA) did not differ among all isolates (79.9, 78.8 and 79.6%; P = 0.86), but decreased for the first isolate per patient (70.2, 65.7, and 64.1%; P = 0.006) over time. Annual duplication rates of methicillin-susceptible S. aureus (MSSA) decreased (39.6, 37.6, and 31.7%; P = 0.01), but those of MRSA increased significantly (64.3, 67.8, and 68.9%; P = 0.004). Rates of cefotaxime-resistant K. pneumoniae did not differ over time by either method, and rates of imipenem-resistant A. baumannii decreased over time by both methods. Duplication rates did not differ for either susceptible or resistant isolates of K. pneumoniae and A. baumannii. The trends in MRSA rate differed by the two methods because of the different proportion of duplicate isolates per year. MRSA rates might be increasingly overestimated for all isolates. These results suggest that the method of calculating results for the first isolate per patient may remove the effect of duplication, allowing the simple and unambiguous analysis of cumulative susceptibility rates.     

Synergic in-vitro activity of imipenem and sulbactam against Acinetobacter baumannii

The interaction between sulbactam and imipenem was evaluated with four clinical isolates of Acinetobacter baumannii, including two isolates resistant to imipenem, one of which produced IMP-1 metallo-beta-lactamase. Two isolates (one of which was imipenem-resistant) were sulbactam-resistant by undefined mechanisms. MICs were determined by standard broth microdilution methods. Time-kill assays with imipenem and sulbactam, alone or in combination at 0.5 x MIC and 1 x MIC, showed a synergic effect in all four isolates of A. baumannii after incubation for 0, 4, 8 and > 24 h at 35 degrees C.     

鲍曼不动杆菌临床株多重药物外排泵AdeABC的研究

目的 研究鲍曼不动杆菌(Acinetobacter baumannii)临床株外排泵AdeABC的表达与耐药的关系及表达调控.方法 微量肉汤稀释法检测鲍曼不动杆菌临床株对抗菌药物的敏感性及泵抑制剂作用,RT-PCR检测泵基因adeB的mRNA表达水平,PCR扩增泵调控基因adeRS并测序分析.结果 30株多重耐药的鲍曼不动杆菌临床株及5株敏感株均存在泵结构基因片段adeB和调控基因adeRS,随机选取15株多重耐药株中均检测到adeB的mRNA表达,而在5株敏感株中无表达.测定2株多重耐药株调控基因adeRS序列均出现基因突变,发生氨基酸替代及缺失.结论 鲍曼不动杆菌临床株外排泵AdeABC的表达可能与耐药性有关,在多重耐药株中存在着调控基因adeRS的基因序列变化.     

Antimicrobial resistance surveillance of bacteria in 1999 in Korea with a special reference to resistance of enterococci to vancomycin and gram-negative bacilli to third generation cephalosporin, imipenem, and fluoroquinolone.

The trend of antimicrobial resistance of bacteria isolated from patients in 30 Korean hospitals in 1999 was analyzed with a particular attention to cefotaxime- or fluoroquinolone-resistant gram-negative bacilli, imipenem-resistant Pseudomonas aeruginosa, and vancomycin-resistant enterococci. Adequacy of susceptibility testing, and any change in the frequencies of isolated species were also analyzed. The results showed that only 20% and 30% of hospitals tested the piperacillin-tazobactam and cefoxitin susceptibility of Enterobacteriaceae, respectively, only 24% of hospitals the piperacillin-tazobactam susceptibility of P. aeruginosa, and 17% of hospitals the fusidic acid susceptibility of staphylococci. Among the isolates 26.3% were glucose-nonfermenting gram-negative bacilli, and 34.7% of Enterococcus were Enterococcus faecium. Slight decline of cefotaxime-resistance rate to 20% was noted in Klebsiella pneumoniae, while fluoroquinolone-resistance rate was 68% in Acinetobacter baumannii. The ceftazidime- and imipenem-resistance rates were 17% and 18%, respectively in P. aeruginosa. The vancomycin-resistance rate of E. faecium rose significantly to 15.1%, but the rates varied significantly depending on hospitals suggesting presence of different degree of selective pressure or nosocomial spread. In conclusion, the prevalence of imipenem-resistant P. aeruginosa and the increase of vancomycin-resistant E. faecium were the particularly worrisome phenomena observed in this study.     

Clonal spread of imipenem-resistant Acinetobacter baumannii among different cities of China

A total of 342 imipenem-resistant Acinetobacter baumannii isolates (IRABs) were collected from 16 Chinese cities. Six predominant clones had spread widely, and four clones were detected in distant hospitals. The majority of the IRABs contain bla(OXA-23), with ISAba1 upstream of the gene. These results suggested that clonal spread played an important role in the outbreak of IRABs in China.     

Comparison of in vitro activities of levofloxacin, ciprofloxacin, ceftazidime, cefepime, imipenem, and piperacillin-tazobactam against aerobic bacterial pathogens from patients with nosocomial infections.

BACKGROUND AND PURPOSE: There is a rapid worldwide emergence of multidrug-resistant pathogens, especially in nosocomial isolates. This study compared the in vitro activities of levofloxacin, ciprofloxacin, ceftazidime, cefepime, imipenem, and piperacillin-tazobactam against 208 aerobic bacterial pathogens that caused 197 nosocomial infections in 184 patients. METHODS: Antimicrobial susceptibility was evaluated by E test in accordance with the guidelines of the National Committee for Clinical Laboratory Standards. RESULTS: Most (140/208, 67%) of the isolates were facultative Gram-negative bacilli. Levofloxacin and ciprofloxacin were the most effective (22/22, 100%) against oxacillin-sensitive Staphylococcus aureus. None of the antibiotics tested were found to be effective (0/25) against oxacillin-resistant S. aureus. Of the 11 isolates of Acinetobacter baumannii that were not pandrug-resistant (PDR), only 9 isolates (9/11, 81%) were sensitive to imipenem and 5 isolates (5/11, 45%) were sensitive to levofloxacin, ciprofloxacin, and ceftazidime. Another 22 isolates of A. baumannii that were PDR were completely resistant to all 6 antibiotics. The majority of isolates of Pseudomonas aeruginosa were sensitive to these 6 antimicrobial agents with 10/11 (91%) sensitive to levofloxacin and ciprofloxacin, 9/11 (83%) sensitive to ceftazidime, cefepime and piperacillin-tazobactam, and 8/11 (75%) sensitive to imipenem. CONCLUSIONS: The majority of the bacterial isolates causing nosocomial infections were found to be sensitive to the 6 antibiotics tested. Bacterial isolates of nosocomial infections that were completely resistant to these 6 antibiotics were PDR A. baumannii, PDR P. aeruginosa, and oxacillin-resistant S. aureus. More potent antimicrobial agents are needed to treat infections caused by PDR A. baumannii and PDR P. aeruginosa.     

醋酸钙-鲍曼不动杆菌复合体的精确鉴定与药敏分析

目的精确鉴定醋酸钙-鲍曼不动杆菌复合体菌株;检测菌株对氨基糖苷类抗生素和碳青霉烯类抗生素的敏感性。方法运用自动化分析仪VITEK 2试卡法对临床分离不动杆菌进行菌种鉴定,对鉴定为醋酸钙-鲍曼不动杆菌复合体的菌株进一步经16S rRNA序列分析确证其准确种属。分别用自动化分析仪VITEK 2和微稀释法测定精确鉴定后的醋酸钙-鲍曼不动杆菌复合体菌株对阿米卡星、庆大霉素、妥布霉素、亚胺培南和厄他培南五种抗生素的敏感性,分析药敏实验结果。结果共进行了232株不动杆菌的VITEK 2鉴定,其中195株鉴定为醋酸钙-鲍曼不动杆菌复合体。对195株醋酸钙-鲍曼不动杆菌复合体菌株进一步用16S rRNA序列分析确证其准确种属,结果显示173株为鲍曼不动杆菌,22株为醋酸钙不动杆菌。对173株鲍曼不动杆菌及22株醋酸钙不动杆菌分别用VITEK 2和微稀释法进行阿米卡星、庆大霉素、妥布霉素、亚胺培南和厄他培南五种抗生素的药敏检测。微稀释法药敏结果显示,受试鲍曼不动杆菌对三种氨基糖苷类抗生素均呈现高水平耐药,而对两种碳青霉烯类抗生素敏感度较高;受试醋酸钙不动杆菌对五种抗生素均呈现较高敏感度。与微稀释法药敏检测结果相比,VITEK 2试卡法药敏结果中受试鲍曼不动杆菌和醋酸钙不动杆菌对各抗生素的药敏检测结果均出现了不同程度的误差,鲍曼不动杆菌药敏检测结果中阿米卡星符合率最低,严重错误率高达34.10%;醋酸钙不动杆菌药敏检测结果中厄他培南符合率最低,重大错误率高达40.91%。结论 VITEK 2在不动杆菌种属鉴定中存在局限性,辅以16S rRNA序列分析,方可精确鉴定醋酸钙-鲍曼不动杆菌复合体。鲍曼不动杆菌对氨基糖苷类抗生素耐药现状严重。用VITEK 2进行鲍曼不动杆菌和醋酸钙不动杆菌对氨基糖苷类和碳青霉烯类的药敏测定时存在不同程度的误差,建议辅以微稀释法。     

Characterization of a hospital outbreak of imipenem-resistant Acinetobacter baumannii by phenotypic and genotypic typing methods.

During a 13-month period, 31 patients hospitalized primarily in two intensive care units (ICUs) were either colonized or infected by imipenem-resistant Acinetobacter baumannii. Typing of the isolates by three methods (antibiotyping, biotyping, and pulsed-field gel electrophoresis) revealed that two distinct strains were involved in the first 9 cases of the outbreak and that one of these strains, which had acquired a higher level of imipenem resistance as well as resistance to all aminoglycosides, accounted for 21 of 22 cases in the second part of the outbreak. ICU environmental contamination was recognized as an important reservoir of this epidemic strain. The outbreak ceased only after the ICUs were closed for complete cleaning and disinfection.     

High prevalence of carbapenem-hydrolysing oxacillinases in epidemiologically related and unrelated Acinetobacter baumannii clinical isolates in Spain

Carbapenem-hydrolysing oxacillinases are reported increasingly in Acinetobacter baumannii. This study investigated the role of these beta-lactamases in causing resistance to carbapenems in 83 epidemiologically related and unrelated imipenem-resistant A. baumannii clinical isolates. The isolates were also analysed for the presence of ISAba1 in the promoter region of the bla(OXA-51)-like gene in order to investigate the role of ISAba1 in OXA-51 expression. All clinical isolates contained a bla(OXA-51)-like gene, 20% contained a bla(OXA-58)-like gene, and 42% contained a bla(OXA-40)-like gene; bla(OXA-23)-like, bla(IMP) and bla(VIM) genes were not detected in any of the isolates investigated. ISAba1 was found in 24 (82.7%) of 28 pulsetypes, and was located in the promoter region of the bla(OXA-51)-like gene in five (20.8%) of these pulsetypes. Expression of bla(OXA-51) was detected in the five isolates with ISAba1 located in the promoter region, but was not detected in an isogenic imipenem-susceptible A. baumannii isolate that did not have ISAba1 located in the promoter region. It was concluded that there is a high prevalence of oxacillinases with activity against carbapenems among genetically unrelated A. baumannii clinical isolates from Spain, and that concomitant expression of two carbapenemases (OXA-51-like and either OXA-40-like or OXA-58-like) may take place. Insertion of an ISAba1-like element in the promoter of the bla(OXA-51)-like gene promotes the expression of this gene, although this did not seem to play a major role in carbapenem resistance.     

Genotypic diversity and antibiotic susceptibility of Acinetobacter baumannii isolates in a Bulgarian hospital

A set of 18 Acinetobacter baumannii isolates, collected prospectively in a Bulgarian hospital during episodes of increased A. baumannii occurrence during 2000-2002, was investigated for genotypic diversity and antibiotic susceptibility. Four genotypes were identified by amplified fragment length polymorphism genomic fingerprinting, one of which (type 1) accounted for 13 isolates, indicating that a specific strain was predominant. The single isolate allocated to type 2 was identified to European clone I. All isolates were resistant to multiple antibiotics, but most retained susceptibility to tobramycin and colistin, and all except one were susceptible to imipenem.     

Identification of epidemic strains of Acinetobacter baumannii by integrase gene PCR

Forty-eight clinical Acinetobacter isolates with different epidemic behavior were investigated for the presence of integrons and plasmids and for antibiotic susceptibility. Integrons were demonstrated in 50% of the strains by an integrase gene PCR. Epidemic strains of Acinetobacter baumannii were found to contain significantly more integrons than nonepidemic strains. Also, the presence of integrons was significantly correlated with simultaneous resistance to several antibiotics. Plasmids were detected in 42% of the strains. However, there was no significant correlation between the numbers of plasmids and integrons in Acinetobacter species strains, no significant difference in the number of plasmids between epidemic and nonepidemic A. baumannii strains, and no significant correlation between the presence of plasmids and antibiotic resistance. Hence, it is likely that integrons play an important role in antibiotic resistance and thereby in the epidemic behavior of A. baumannii. Because the integrase gene PCR identified almost three-quarters of the epidemic A. baumannii isolates (17 of 23), this seems to be a rapid and simple technique for the routine screening and identification of clinical A. baumannii isolates with epidemic potential.     

Clinical and molecular epidemiology of an outbreak of infusion-related Acinetobacter baumannii bacteremia in an Intensive Care Unit

Objective: To describe the clinical and molecular epidemiology of an outbreak of infusion-related Acinetobacter baumannii bacteremia in an intensive care unit (ICU). Methods: Six cases of A. baumannii bacteremia identified in the Foligno Hospital ICU, Italy, were peer reviewed. Antibiotic susceptibility and genotyping (PFGE and RAPD) of A. baumannii isolates were carried out. Results: All A. baumannii blood isolates and a strain isolated from parenteral solution had an identical genotype. The strains were susceptible to carbapenems and the combination of meropenem plus amikacin or piperacillin/ tazobactam plus netilmicin was synergistic. A. baumannii bacteremia persisted for several days in almost all patients; catheter tip cultures were always positive for A. baumannii. Three patients, with an elevated Apache II score, died of sepsis. Conclusions: The outbreak was related to contaminated parenteral solutions improperly prepared in the ward. Aseptic preparation in the hospital pharmacy allowed for an interruption of the outbreak.     

Oligonucleotide array-based identification of species in the Acinetobacter calcoaceticus-A. baumannii complex in isolates from blood cultures and antimicrobial susceptibility testing of the isolates

Acinetobacter calcoaceticus, A. baumannii, Acinetobacter genomic species (gen. sp.) 3, and Acinetobacter gen. sp. 13TU, which are included in the A. calcoaceticus-A. baumannii complex, are difficult to distinguish by phenotypic methods. An array with six oligonucleotide probes based on the 16S-23S rRNA gene intergenic spacer (ITS) region was developed to differentiate species in the A. calcoaceticus-A. baumannii complex. Validation of the array with a reference collection of 52 strains of the A. calcoaceticus-A. baumannii complex and 137 strains of other species resulted in an identification sensitivity and specificity of 100%. By using the array, the species distribution of 291 isolates of the A. calcoaceticus-A. baumannii complex from patients with bacteremia were determined to be A. baumannii (221 strains [75.9%]), Acinetobacter gen. sp. 3 (67 strains [23.0%]), Acinetobacter gen. sp. 13TU (2 strains [0.7%]), and unidentified Acinetobacter sp. (1 strain [0.3%]). The identification accuracy of the array for 12 randomly selected isolates from patients with bacteremia was further confirmed by sequence analyses of the ITS region and the 16S rRNA gene. Antimicrobial susceptibility testing of the 291 isolates from patients with bacteremia revealed that A. baumannii strains were less susceptible to antimicrobial agents than Acinetobacter gen. sp. 3. All Acinetobacter gen. sp. 3 strains were susceptible to ampicillin-sulbactam, imipenem, and meropenem; but only 67.4%, 90%, and 86% of the A. baumannii strains were susceptible to ampicillin-sulbactam, imipenem, and meropenem, respectively. The observed significant variations in antimicrobial susceptibility among different species in the A. calcoaceticus-A. baumannii complex emphasize that the differentiation of species within the complex is relevant from a clinical-epidemiological point of view.     

鲍曼不动杆菌的临床分布特征及耐药趋势分析

监测医院近年临床标本中鲍曼不动杆菌的分布特点及其耐药情况,以指导临床合理用药。将2004年1月至2008年12月从主要来源于呼吸内科（23.0%）、老年病科（20.0%）、ICU中心（9.9%）、血液科（8.9%）等科室患者的不同标本中分离的鲍曼不动杆菌,采用CLSI/NCCLS标准K-B法分析其对临床常用抗菌药物的耐药情况,用Whonet5.4软件进行统计学分析。结果如下：2004至2008年鲍曼不动杆菌检出率分别为4.4%、4.0%、4.6%、4.6%、4.7%,痰中的检出率最高,为84.1%。该菌耐药现象严重,约60%对哌拉西林、头孢噻肟、氨曲南等呈多重耐药。敏感度最高的是碳青霉烯类,约84.0%。     

IMPs, VIMs and SPMs: the diversity of metallo-β-lactamases produced by carbapenem-resistant Pseudomonas aeruginosa in a Brazilian hospital

Pseudomonas aeruginosa isolates (n=183), collected from bacteraemic patients hospitalised in Sao Paulo Hospital (Brazil) during 2000-2001, were screened for susceptibility to antimicrobial agents. The polymyxins were the most active compounds (100% susceptibility), followed by amikacin and cefepime (59.0%), meropenem (57.4%), and imipenem and gentamicin (55.2%). Imipenem-resistant isolates were ribotyped and screened for production of metallo-beta-lactamases (MBLs) by PCR with primers for bla(IMP), bla(VIM) and bla(SPM). MBL production was detected in 36 isolates (19.7% of the entire collection; 43.9% of the imipenem-resistant isolates) and the MBLs included SPM-1-like (55.6%), VIM-2-like (30.6%) and IMP-1-like (8.3%) enzymes.     

Comparison of four different methods for epidemiologic typing of Acinetobacter baumannii.

A set of 103 epidemiologically well-defined Acinetobacter baumannii isolates obtained from nine hospital outbreaks and 21 unrelated strains were characterized by pulsed-field gel electrophoresis (PFGE) of total genomic DNA digested with ApaI. Among outbreak strains, eight different patterns and five possible variants were identified by PFGE. Results were compared with those from traditional typing methods such as plasmid profile analysis, antimicrobial susceptibility, and biotyping. Plasmid analysis revealed six different and two related patterns; one outbreak strain lacked plasmids. A total of 16 of the 21 unrelated strains harbored plasmids and exhibited unique patterns. Epidemiologically unrelated strains were placed into only two biotypes and had similar antimicrobial susceptibility patterns but were clearly distinguished by PFGE. PFGE of A. baumannii chromosomal DNA yielded reproducible and easily readable results and showed excellent discriminatory power. However, plasmid profile analysis may provide a cost-effective first step in epidemiological typing of A. baumannii isolates obtained from well-defined hospital outbreaks.     

Activity of meropenem against imipenem-resistant bacteria and selection in vitro of carbapenem-resistantEnterobacteriaceae

The activity of meropenem against 106 imipenem-resistant (MIC 8 mg/l) clinical isolates, and the frequency of resistance to meropenem and imipenem among 24Enterobacteriaceae was determined. Both agents selected colonies on agar but 20–80 % were susceptible after one subculture and 72 % of the mutants reverted to susceptibility 1 to 6 months after selection. All isolates and stable mutants were inhibited by > 1 mg/l meropenem, although the MIC of imipenem was 4–16 mg/l. Three of sixXanthomonas maltophilia isolates were susceptible to meropenem (MICs 2–4 mg/l).Pseudomonas aeruginosa lacking outer membrane protein D2 were resistant to meropenem, although isolates with substantially reduced expression of this protein were susceptible. None of the imipenem-resistant gram-positive bacteria were susceptible to meropenem. There was no clear correlation between altered outer membrane protein expression and decreased susceptibility to carbapenems, and there was no apparent involvement of plasmid or chromosomal -lactamase.