某院2011年重症监护病房鲍曼不动杆菌感染的耐药性分析

目的了解某院2011年重症监护病房（ICU）内鲍曼不动杆菌的耐药情况,为预防与治疗鲍曼不动杆菌感染提供依据。方法回顾性调查某院2011年ICU35例鲍曼不动杆菌感染病例的标本分布、耐药性、危险因素和抗菌药物使用情况。结果 ICU鲍曼不动杆菌主要来源于痰标本（35株,占85.37%）,其次为创面分泌物（3株,占7.31%）。有基础疾病、侵入性操作、前期使用抗菌药物和激素、入住ICU、住院时间大于60d等是感染鲍曼不动杆菌的高危因素。鲍曼不动杆菌对头孢哌酮舒巴坦（100.0%）及亚胺培南西司他汀（92.5%）的敏感率较高,对其他16种抗菌药物的耐药率均在50.0%以上。临床治疗鲍曼不动杆菌感染多经验性选择第三代头孢菌素、喹诺酮及广谱青霉素类抗菌药物。结论 ICU内鲍曼不动杆菌耐药情况严重,控制感染的关键在于注意高危因素的防范,加强对该菌的耐药监测,合理使用抗菌药物。     

Epidemiology of Acinetobacter spp.-associated healthcare infections and colonization among children at a tertiary-care hospital in Saud Arabia: a 6-year retrospective cohort study

A retrospective cohort study was conducted among hospitalized children less than 12?years of age who had Acinetobacter spp. isolated from ≥1 cultures between October 2001 and December 2007 at King Abdulaziz Medical City in Riyadh, Saudi Arabia. Children with multidrug-resistant (MDR) Acinetobacter spp. healthcare-associated infections (HAIs) were compared to children with antimicrobial-susceptible Acinetobacter spp. HAIs and to children colonized with Acinetobacter. Children with MDR Acinetobacter spp. HAIs were older (p?=?0.01), more likely to be admitted to an intensive care unit (ICU) (p?=?0.06), and had a higher mortality rate (p?=?0.02) than colonized children. Children with MDR Acinetobacter spp. HAIs were older than children with antimicrobial-susceptible Acinetobacter spp. HAIs (p?=?0.0004), but their mortality rates were similar. Among children with MDR Acinetobacter spp. HAIs, burn injuries were the most common underlying illness. HAIs caused by MDR or susceptible Acinetobacter spp. occurred after prolonged hospitalization, suggesting nosocomial acquisition. Patients infected with MDR Acinetobacter spp. frequently received inappropriate empiric therapy (73.9?%). Further studies are needed in order to identify effective strategies to prevent nosocomial transmission and effective ways of improving patient outcomes.     

Community-acquired bacteremic Acinetobacter pneumonia in tropical Australia is caused by diverse strains of Acinetobacter baumannii, with carriage in the throat in at-risk groups

Acinetobacter isolates from eight subjects with community-acquired Acinetobacter pneumonia (CAAP), a major cause of fatal community-acquired pneumonia in tropical Australia, were phenotypically and genotypically confirmed by pulsed-field gel electrophoresis analysis to be broadly diverse Acinetobacter baumannii strains. Wet-season throat carriage of A. baumannii was found in 10% of community residents with excess levels of alcohol consumption, the major at-risk group for CAAP.     

Glucose dehydrogenase activity in Acinetobacter species

A study of D-glucose oxidation by Acinetobacter species was carried out. Glucose-oxidizing strains were found distributed among almost all Acinetobacter species. 14C-glucose oxidation kinetics by non-proliferating cells with separation of oxidation products (14C-gluconate) by DEAE-cellulose paper chromatography was studied. Inhibition of glucose dehydrogenase (GDH) activity by 11 carbohydrates (mono- and disaccharides) and determination of the kinetic parameters showed that glucose oxidation was due to the action of membrane-bound GDH (inactive in vivo on disaccharides). On the basis of GDH inhibition patterns obtained, two groups were individualized. The first group of strains (identified as A. calcoaceticus, A. baumannii, A. lwoffii, A. johnsonii and Acinetobacter species 3, 9, 10 and 11) showed a greater affinity for glucose than the second group (A. haemolyticus, A. junii and Acinetobacter species 6 and 12). Restoration of GDH activity after addition of pyrroloquinoline quinone (PQQ) was studied in 187 strains previously found unable to oxidize glucose. GDH activity of 150 out of 166 strains identified as A. baumannii, A. johnsonii, A. lwoffii and Acinetobacter species 11 and 12 was restored. Eighteen of 21 strains identified as A. haemolyticus and Acinetobacter species 6 were unable to produce acid from glucose after addition of PQQ. Our results confirm that the former taxonomic scheme for the genus Acinetobacter (2 species differing only by glucose oxidation) is untenable and that, accordingly, identification of Acinetobacter strains at the species level must be performed using more modern methods, i.e. carbon source utilization tests.     

Antimicrobial resistance of Acinetobacter spp. in Europe

Bacteria of the genus Acinetobacter are ubiquitous in nature. These organisms were invariably susceptible to many antibiotics in the 1970s. Since that time, acinetobacters have emerged as multiresistant opportunistic nosocomial pathogens. The taxonomy of the genus Acinetobacter underwent extensive revision in the mid-1980s, and at least 32 named and unnamed species have now been described. Of these, Acinetobacter baumannii and the closely related unnamed genomic species 3 and 13 sensu Tjernberg and Ursing (13TU) are the most relevant clinically. Multiresistant strains of these species causing bacteraemia, pneumonia, meningitis, urinary tract infections and surgical wound infections have been isolated from hospitalised patients worldwide. This review provides an overview of the antimicrobial susceptibilities of Acinetobacter spp. in Europe, as well as the main mechanisms of antimicrobial resistance, and summarises the remaining treatment options for multiresistant Acinetobacter infections.     

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。     

EndemicAcinetobacter anitratus in a surgical intensive care unit: Mechanical ventilators as reservoir

Colonization and infection with an endemic multiresistant strain ofAcinetobacter calcoaceticus varietyanitratus had been observed in the surgical intensive care unit of a university hospital since 1982. An outbreak of infection with this endemic, multiresistantAcinetobacter anitratus strain occurred between January and September, 1984. After initial attempts at identification of environmental reservoirs had been unsuccessful, risk factors for the acquisition ofAcinetobacter anitratus were investigated by comparing the epidemiological characteristics of patients who became colonized or infected with those of control patients without colonization. The results of this case-control study and of the ensuing specific cultures indicated that ventilators in use in the unit were the reservoirs ofAcinetobacter anitratus, resulting in frequent nosocomial respiratory tract infections. After modification of the mechanical ventilators, colonization and infection rates withAcinetobacter anitratus decreased. Since January 1985, no new cases of colonization or infection with this endemic strain ofAcinetobacter anitratus have been recorded.     

Epidemiological typing and prevalence of integrons in multiresistant Acinetobacter strains

Seventy-seven Acinetobacter isolates were recovered from patients in a Korean hospital during the period from November to December 1998. The isolates were genotyped using randomly amplified polymorphic DNA (RAPD) analysis for epidemiological relationship, and investigated for antibiotic susceptibility and presence of integrons. Sixty-nine Acinetobacter baumannii isolates were distributed into five groups by RAPD profiles, with 5, 1, 60, 2 and 1 in each group. The major RAPD group of 60 isolates was further divided into six subgroups by antibiograms. Eight isolates belonging to Acinetobacter DNA group 13TU were distributed among six RAPD groups. Seventy-three of the Acinetobacter isolates were resistant to eight or more of the antibiotics tested. Integrase genes were detected in 66 of the 69 A. baumannii (96%) and in 5 of the 8 Acinetobacter DNA group 13TU isolates (63%). The intI1 and intI2 genes were found in 63 and 8 isolates, respectively. The intI3 gene was not detected. All integron-carrying isolates were resistant to multiple antibiotics. All strains isolated from more than one patient carried integrons. According to the results, the presence of integrons was significantly (p<0.01) associated with multiple antibiotic resistance and nosocomial spread in Acinetobacter strains.     

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.     

Multiresistant Acinetobacter infections: a role for sulbactam combinations in overcoming an emerging worldwide problem

Recent studies have highlighted the emergence of infections involving multiresistant Acinetobacter clinical isolates. Sulbactam offers direct antimicrobial activity against Acinetobacter species. Accordingly, co-administration of sulbactam with ampicillin or cefoperazone offers the potential of effective empirical therapy against Acinetobacter and other bacteria such as Enterobacteriaceae in institutions in which they are susceptible. Many in vitro studies have indicated that Acinetobacter remains fully susceptible to ampicillin–sulbactam or cefoperazone–sulbactam. Furthermore, ampicillin–sulbactam has proven clinically effective and well tolerated in the treatment of severe acinetobacter infections, including bacteremia. Therefore, ampicillin–sulbactam is a sensible option for the treatment of life-threatening acinetobacter infections.     

Species distribution and physiological characterization of Acinetobacter genospecies from healthy human skin of tribal population in India

BACKGROUND: Various reports on distribution of Acinetobacter spp. from healthy human skin restricted to urban population. However, no such data is available from healthy human skin of tribal population not exposed to modern antibiotics during their life time. PURPOSE: Isolation, biotyping, distribution and physiological characterisation of Acinetobacter spp. from healthy human skin of tribal population. METHODS: Tribal population of Toranmal area of Satpuda Ranges, Maharashtra, India were sampled for ten body sites. Tentative Acinetobacter isolates were confirmed to the genus level by chromosomal DNA transformation assay and to species level using Bouvet and Grimont system. Novel physiological characteristics like pH, temperature and salt tolerance were studied. All strains were screened for production of various enzymes. RESULTS: One hundred and eighteen strains were isolated, which belonged to nine Acinetobacter genospecies. A. haemolyticus was most abundant followed by A. calcoaceticus and A. genospecies 1-3. Higher percentage of Acinetobacter was recovered from skin of nose, Pawara tribe and female volunteers. They showed wide variation in temperature, salt and pH tolerance. Most of the strains could produce enzymes viz, lipase, esterase, urease and amylase. CONCLUSIONS: Acinetobacter spp. belonging to nine genospecies were obtained in the present study. Physiological characteristics including high salt, temperature and acidic pH tolerance were helpful to differentiate between the commensal and pathogenic species of Acinetobacter genus.     

Clinical impact and pathogenicity of Acinetobacter

Members of the genus Acinetobacter have been implicated in a wide spectrum of infectious diseases. Although this organism is associated primarily with nosocomial infections, it has also been involved in cases of community-acquired infection. Before the 1970s, Acinetobacter infections were mostly post-surgical urinary tract infections in patients hospitalised in surgical units. The significant improvement in resuscitation techniques during the last 30 years has changed the types of infection caused by Acinetobacter. Since the 1980s, Acinetobacter has spread rapidly among patients in intensive care units. Today, Acinetobacter accounts for c. 9% of nosocomial infections, with most Acinetobacter infections involving the respiratory tract. Transmission via the hands of hospital staff has become the most important contributory factor in patient colonisation. Acinetobacter baumannii is the species that is involved most frequently in infections of humans, but a natural reservoir for A. baumannii outside the hospital environment has not yet been identified. Community-acquired infection and infections acquired following war or natural disasters (e.g., earthquakes) have been described. Acinetobacter causes mild-to-severe illness, but can be fatal. The severity of Acinetobacter infection depends upon the site of infection and the patient's susceptibility to infection as a result of underlying disease. The circumstances that allow Acinetobacter to assume a pathogenic role are not really well-understood. As this organism is a low-grade pathogen, the pathogenesis of Acinetobacter infections probably involves numerous factors, including virulence determinants, which have yet to be investigated.     

A PCR-based method to differentiate between Acinetobacter baumannii and Acinetobacter genomic species 13TU

A new PCR-based method that exploits differences in gyrB gene sequences was developed to distinguish between Acinetobacter baumannii and Acinetobacter genomic sp. 13TU. Among 118 clinical and reference Acinetobacter strains, 102 of which were previously speciated by amplified rDNA restriction analysis as belonging to the Acinetobacter calcoaceticus-A. baumannii complex, the method correctly identified 31 A. baumannii and 54 Acinetobacter genomic sp. 13TU isolates to the species level. The method was rapid, specific and easy to interpret.     

gyrB multiplex PCR to differentiate between Acinetobacter calcoaceticus and Acinetobacter genomic species 3

A previously established multiplex PCR that identifies to the species level Acinetobacter baumannii and Acinetobacter genomic species 13TU (GS13TU) was expanded to include Acinetobacter calcoaceticus and Acinetobacter genomic species 3.     

Distinct antimicrobial resistance patterns and antimicrobial resistance-harboring genes according to genomic species of Acinetobacter isolates

Using 58 isolates of Acinetobacter species recovered from a university hospital between August 2004 and March 2005, we performed genomic identification by amplified rRNA gene restriction analysis (ARDRA) and investigated the existence of metallo-beta-lactamase (MBL) producers and extended-spectrum beta-lactamase (ESBL) producers. Genomic species identification of Acinetobacter strains using ARDRA showed that 40 strains were genomic species 2 (Acinetobacter baumannii), 9 were 13 sensu Tjernberg and Ursing (13TU), 5 were Acinetobacter phenon 6/ct 13TU, and 4 were Acinetobacter genospecies 3. Among 58 strains, 13 isolates were MBL producers carrying bla(IMP-1) or bla(VIM-2) and 13 isolates were ESBL producers carrying bla(PER-1). Notably, the MBL producers were mostly 13TU, Acinetobacter phenon 6/ct 13TU, and Acinetobacter genospecies 3, which showed susceptibility to ciprofloxacin and ampicillin-sulbactam. However, 12 of 13 strains carrying bla(PER-1) were A. baumannii, showing multidrug resistance. The data revealed that the antimicrobial resistance patterns and resistance-harboring genes of Acinetobacter species are remarkably distinct according to the genomic species of Acinetobacter isolates.     

鲍曼不动杆菌的临床分布及耐药性变迁

目的了解鲍曼不动杆菌在临床标本的分离率和病区分布及耐药性变化趋势。方法菌株鉴定采用法国梅里埃VITEK32细菌鉴定系统进行鉴定,药敏试验采用K-B法,药敏试验结果判定以CLSI/NCCLS标准进行。结果 2004-2010年共收到26670份标本,分离出阳性细菌7065株,其中鲍曼不动杆菌471株（6.67%）,分离率为1.77%（471/7065）。在471株鲍曼不动杆菌中,从痰液标本分离出最多有409株（86.83%）,分离率最高的是ICU,占49.3%。鲍曼不动杆菌对抗菌药物的耐药性普遍较高,且呈逐年升高趋势,部分表现出多重耐药特征。结论鲍曼不动杆菌的耐药状况日益严重,应重视临床鲍曼不动杆菌的感染与分离,谨防多重耐药鲍曼不动杆菌的院内感染及暴发流行。     

Detection of integrons in worldwide nosocomial isolates of Acinetobacter spp.

Objective: To examine the distribution of integrons in genotypically unrelated worldwide multiresistant clinical isolates of Acinetobacter spp.
Methods: The presence and genetic location of class 1, 2 and 3 integrons were examined in a genotypically heterogeneous collection of 25 nosocomial isolates of Acinetobacter spp., from 15 locations in 11 different countries worldwide, by hybridization and PCR-based methods. Class 1 integron structures were characterized genetically by a PCR mapping technique.
Results: Class 1 integrons were detected in 17 of the 25 Acinetobacter isolates examined. Only one isolate contained a class 2 integron. No class 3 integrons were detected. The integrons varied in size and in the number of inserted cassettes, but similar integrons were found in genotypically distinct isolates from different locations worldwide. These structures were integrated into the chromosome in all isolates where they were detected, although some integrons were capable of subsequent transfer or mobilization. Genes coding for aminoglycoside-modifying enzymes formed the predominant cassettes identified within the integrons.
Conclusions: Clinical isolates of Acinetobacter spp. from diverse locations seem to share resistance mechanisms acquired from other genera by a variety of mechanisms, including dissemination of integrons. Once integrons are incorporated into the bacterial genome, Acinetobacter spp. are potentially able to act as a reservoir of resistance genes for other species and genera.     

Prevalence of Acinetobacter baumannii and other Acinetobacter spp. in faecal samples from non-hospitalised individuals

In total, 226 individuals from the community were investigated for faecal carriage of Acinetobacter spp. by broth enrichment culture, followed by growth on blood agar and/or Leeds Acinetobacter Medium (LAM). Acinetobacter baumannii was isolated on both LAM and blood agar from one of 100 specimens in the UK and one of 126 specimens in The Netherlands. The predominant species were Acinetobactor johnsonii and genomic sp. 11, which were cultured from 22 and five specimens, respectively. A. baumannii did not seem to be widespread in the faecal flora of individuals in the community.     

2007-2009年不动杆菌检出情况及耐药性分析

目的了解近三年不动杆菌的临床分布及耐药性变迁情况,为临床合理用药提供依据。方法采用回顾性调查方法 ,对2007-2009年临床分离的不动杆菌的临床分布及耐药性变迁情况进行统计分析,菌株鉴定和药物敏感试验方法采用法国梅里埃WITEK2系统并分别用K-B法和E-test法测定48株不动杆菌对米诺环素和多粘菌素的敏感性。结果 230株不动杆菌中鲍曼不动杆菌所占比例最高,为93%（214/230）。主要分离自痰标本（203/230）,以呼吸内科和ICU最多见,分别占36.5%和14.3%。2007-2009年不动杆菌年分离率分别为5.1%（53/1039）、6.4%（81/1266）和7.3%（96/1307）,呈逐年增高的趋势,耐药率较低的有亚胺培南5.7%（2009年）、阿米卡星9.2%（2009年）,耐药率较高的有环丙沙星50.6%（2009年）和复方新诺明47.1%（2009年）,而且耐药率有逐年上升趋势。多重耐药菌株分离率2007年为11.3%（6/53）,2008年为24.7%（20/81）,2009年为35.4%（34/96）,其中泛耐药株2007年0株,2008年2（2.5%）株,2009年4（4.2%）株。48株不动杆菌对米诺环素敏感率为52.1%,多粘菌素敏感率为100%。结论本院分离的不动杆菌以鲍曼不动杆菌为主,大多数来自痰标本,主要来源于呼吸内科和ICU,分离率有逐年增高的趋势,且耐药率、多重耐药株及泛耐药株均有上升趋势,值得临床高度重视。     

Plasmid DNA fingerprinting of Acinetobacter species other than Acinetobacter baumannii.

During the last years Acinetobacter species have emerged as clinically significant pathogens. Most infections are nosocomially acquired and mainly due to Acinetobacter baumannii. Little is known about the epidemiology and clinical significance of unnamed Acinetobacter species 3 (the second most often encountered member of the genus Acinetobacter) and other Acinetobacter species such as A. johnsonii, A. junii, and A. lwoffii. Seventy-five clinical isolates of Acinetobacter species other than A. baumannii (Acinetobacter species 3, n = 37; A. johnsonii, n = 20; A. junii, n = 8; A. lwoffii, n = 10) recovered from 66 patients over a period of 12 months were analyzed by plasmid DNA fingerprinting. Plasmids were found in 84.4% of Acinetobacter species 3 isolates and in all A. johnsonii, A. junii, and A. lwoffii isolates. Strains harbored up to 15 plasmids each. Almost every isolate gave a unique plasmid pattern. With one exception, identical plasmid profiles were detected only in corresponding isolates recovered from blood cultures and intravascular catheters from a given patient. Plasmid DNA fingerprinting proved to be useful for typing Acinetobacter species other than A. baumannii. There was no evidence of patient-to-patient transmission or hospital outbreaks due to these species. This finding is in contrast to the results obtained in studies of the hospital epidemiology of A. baumannii.