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A comparative study of different PCR-based DNA fingerprinting techniques for typing of the Acinetobacter calcoaceticus-A. baumannii complex.

Different PCR-based DNA fingerprinting techniques were evaluated for typing 26 clinical isolates belonging to the Acinetobacter calcoaceticus-A. baumannii complex. Seven isolates belonged to a previously defined outbreak while 19 isolates were unrelated epidemiologically. The PCR-based DNA fingerprinting techniques used were: (i) repetitive extragenic palindromic (REP) PCR; (ii) enterobacterial repetitive intergenic consensus (ERIC) PCR; (iii) randomly amplified polymorphic DNA with M13 forward primer; (iv) restriction analysis of the amplified 16S rRNA gene (ARDRA-16S); and (v) restriction analysis of an amplified region containing the 16S-23S rRNA spacer region and part of the 23S rRNA gene (ARDRA 23S + spacer). The discrimination index for the PCR-based DNA fingerprinting techniques was: 0.99 for REP; 0.94 for ERIC; 0.87 for M13; 0.60 for ARDRA-16S digested with Hpa II and <0.50 for ARDRA 23S + spacer. It was concluded that REP-PCR possessed high discriminatory power and reproducibility in comparison with the other PCR-based DNA fingerprinting techniques, and is a simple and rapid typing method for use in epidemiological studies of isolates belonging to the A. calcoaceticus-A. baumannii complex.

Acinetobacter↗

Typing and characterization of carbapenem-resistant Acinetobacter calcoaceticus-baumannii complex in a Chinese hospital.

This study was designed to investigate the prevalence of carbapenem-resistant Acinetobacter calcoaceticus-baumannii complex (Acb complex) and to type carbapenemases. The relatedness of 45 isolates of carbapenem-resistant Acb complex collected from a clinical setting was analysed by PFGE. The carbapenemases produced by these isolates were typed by IEF, a three-dimensional test, 2-mercaptopropanoic acid inhibition assay, PCR and DNA cloning and sequencing. Results showed that all 45 isolates were resistant to multiple antibiotics including meropenem. The resistance rates to cefoperazone/sulbactam and ampicillin/sulbactam were 2.2 and 6.5%, respectively. About 71.7-78.3% of these isolates were intermediately resistant to cefepime, ceftazidime and cefotaxime. Forty-five isolates were classified into type A (98%) and B (2%) based on their PFGE patterns. Most of type A isolates were from the ICU. Type A was the dominant isolate, including subtypes A1 (22%), A2 (71%), A3 (2%) and A4 (2%). Only one isolate, from the haematology department, belonged to type B. Forty-three isolates (96%) were positive for carbapenemase. One isolate had two bands by IEF, the pIs of which were 6.64 and 7.17. The band with the pI of 6.64 was OXA-23. The other 42 isolates produced two bands with pIs of 6.40 and 7.01 which could not be inhibited by clavulanic acid, cloxacillin or 2-mercaptopropanoic acid. It can be concluded that the prevalent carbapenem-resistant Acb complex isolates from this hospital all had similar beta-lactamase patterns.

Acinetobacter Infections↗

Emergence of Acinetobacter soli harboring three carbapenemase-encoding genes (blaNDM-1, blaIMP-14, and blaOXA-58) on a single plasmid in an ICU patient.

Acinetobacter soli is an environmentally adaptable species increasingly recognized as an emerging pathogen in hospital settings, particularly in intensive care units (ICUs). In this study, we report the first A. soli isolate from an ICU patient that co-harbors three carbapenemase-encoding genes (blaNDM-1, blaIMP-14, and blaOXA-58) on a single plasmid. Whole-genome sequencing revealed that multidrug resistance in this strain is mediated by a 294,790 bp plasmid, pSLAB-A, carrying 16 antimicrobial resistance genes, including all three carbapenemases. Comparative plasmid analysis showed a highly conserved backbone but identified a unique ~40 kb multidrug-resistance region containing blaNDM-1, blaIMP-14, and eight additional resistance genes. Genetic context analysis indicated that insertion sequences (ISAba125 and ISAba3) and class 1 integrons contribute to the mobilization and accumulation of carbapenemase-encoding genes. Plasmid stability assays demonstrated that pSLAB-A remained stably maintained for more than 90 generations without antibiotic selection. A global survey of the NCBI database identified 15 A. soli strains carrying carbapenemase-encoding genes, most of which were isolated from China, with clinical specimens representing the predominant source. Seven carbapenemase-encoding genes were detected, with blaNDM-1 being the most prevalent. Among eight isolates with complete genomes, all carried carbapenemase-encoding genes on plasmids. Phylogenetic analysis revealed regional dissemination of a clonal lineage across hospitals in Zhejiang Province and sustained nosocomial transmission within a hospital in Taiwan. These findings suggest that the spread of carbapenem resistance in A. soli is largely driven by multidrug-resistance plasmids, facilitating clonal expansion in hospital environments and posing a growing challenge for antimicrobial therapy and infection control in ICUs.IMPORTANCECarbapenem-resistant A. soli is an emerging clinical concern, capable of causing severe invasive infections, including bacteremia, in intensive care unit settings, and its emergence poses substantial challenges to antimicrobial therapy. In this study, we demonstrate that carbapenem resistance in A. soli is predominantly mediated by the acquisition of multidrug-resistance plasmids carrying carbapenemase-encoding genes. Owing to its strong environmental persistence, A. soli can readily undergo nosocomial clonal dissemination once carbapenem resistance is acquired. Moreover, the spread of multidrug plasmids co-harboring multiple carbapenemase-encoding genes may accelerate the evolutionary trajectory of resistance in A. soli, further exacerbating the threat to clinical management. Given its demonstrated capacity to cause hospital-associated infections and to rapidly acquire multidrug resistance, A. soli warrants heightened vigilance from both clinical and public health perspectives.

beta-Lactamases↗

Citywide clonal outbreak of multiresistant Acinetobacter baumannii and Pseudomonas aeruginosa in Brooklyn, NY: the preantibiotic era has returned.

BACKGROUND: Carbapenems are important agents for treating nosocomial gram-negative infections. Carbapenem-resistant bacteria have become increasingly problematic in certain regions. This study determined the citywide prevalence and molecular epidemiological features of carbapenem-resistant Acinetobacter baumannii and Pseudomonas aeruginosa in Brooklyn, NY. METHODS: All unique patient isolates of A baumannii and P aeruginosa were collected from 15 Brooklyn hospitals from July 1, 1999, through September 30, 1999. Antibiotic susceptibilities, the genetic relatedness of resistant isolates, and the relationship between antibiotic use and resistance rates were determined. RESULTS: A total of 419 isolates of A baumannii and 823 isolates of P aeruginosa were collected. For A baumannii, 53% were resistant to meropenem and/or imipenem, and 12% were resistant to all standard antibiotics. Ribotyping revealed that a single clone accounted for 62% of the samples and was isolated from patients at all 15 hospitals. The rate of carbapenem resistance was associated with cephalosporin use at each hospital (P =.004). For P aeruginosa, 24% were resistant to imipenem, 5% to amikacin, and 15% to 29% to other antipseudomonal agents. Ribotyping revealed that 3 clones accounted for nearly half of the isolates and were shared by most hospitals. CONCLUSIONS: Approximately 400 patients were infected or colonized with carbapenem-resistant A baumannii and P aeruginosa during a 3-month period in 1999. A few strains have spread widely throughout hospitals in this region. The prevalence of resistant A baumannii seems to be correlated with cephalosporin use. Multiresistant hospital-acquired bacteria should be viewed as a serious public health issue rather than an individual hospital's problem. An intensive coordinated effort will be needed to effectively address this problem.

Acinetobacter↗

An outbreak of multidrug-resistant Acinetobacter baumannii associated with pulsatile lavage wound treatment.

CONTEXT: Pulsatile lavage is a high-pressure irrigation treatment used increasingly in a variety of health care settings to debride wounds. Infection control precautions are not routinely used during the procedure and are not included in pulsatile lavage equipment package labeling. OBJECTIVES: To investigate an outbreak of multidrug-resistant Acinetobacter baumannii and to test the hypothesis that pulsatile lavage wound treatment was the mode of transmission for the organism. DESIGN: Outbreak case-control investigation including case identification, review of medical records, environmental cultures, and pulsed-field gel electrophoresis. SETTING: A 1000-bed tertiary care hospital in Baltimore, Md, during September and October 2003. PATIENTS: The investigation included 11 patients infected or colonized with multidrug-resistant A baumannii. Seven of these patients met the case definition for the case-control study and were compared with 28 controls randomly selected from a list of inpatients without multidrug-resistant A baumannii who had a wound care consultation. MAIN OUTCOME MEASURE: Infection or colonization with multidrug-resistant A baumannii. RESULTS: Eleven patients had cultures that grew multidrug-resistant A baumannii during the outbreak period. Of the 10 health care-associated cases, 8 had received pulsatile lavage treatment. One strain of multidrug-resistant A baumannii was recovered from all 6 pulsatile lavage patients who had isolates available for pulsed-field gel electrophoresis analysis and from multiple surfaces in the wound care area. Six of 7 cases (86%) were treated with pulsatile lavage vs 4 of 28 controls (14%) (odds ratio, 36; 95% confidence interval, 2.8-1721; P<.001). These results confirm that pulsatile lavage was a significant risk factor for acquisition of multidrug-resistant A baumannii. CONCLUSIONS: Transmission was apparently caused by dissemination of multidrug-resistant A baumannii during the pulsatile lavage procedure, resulting in environmental contamination. Appropriate infection control precautions should be used during pulsatile lavage therapy and should be included in pulsatile lavage equipment labeling.

Acinetobacter Infections↗

Evaluation of SDS-polyacrylamide gel systems for the study of outer membrane protein profiles of clinical strains of Acinetobacter baumannii.

The outer membrane protein (OMP) profiles of 23 blood isolates of Acinetobacter baumannii representing all the different antimicrobial susceptibility patterns observed during a 3-year period in a Spanish hospital were studied. OMPs extracted from envelopes of sonicated cells after solubilisation with 2% of N-lauryl-sarcosinate were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) using the Laemmli's buffers. Eight running gel systems differing in the concentration of polyacrylamide (8%, 10% and 12%) and in the absence or presence of urea (4 M and 6 M) were used in a preliminary study analysing the OMP profiles of four clonally unrelated strains of A. baumannii. When this study was completed, the OMPs of the 23 A. baumannii were analysed in 10% SDS-polyacrylamide gels with 6 M urea and in 12% SDS-polyacrylamide gels. Ten OMP profiles were observed in 10% SDS-polyacrylamide gels with 6 M urea, whereas only 5 OMP profiles were visualised using 12% SDS-polyacrylamide gels. The OMP profiles obtained in 10% SDS-polyacrylamide gels with 6 M urea only partially correlated with those observed in 12% SDS-polyacrylamide gels. In conclusion, the use of 10% SDS-polyacrylamide gels with 6 M urea is recommended for the study of OMP profiles of A. baumannii.

Acinetobacter Infections↗

Acinetobacter baumannii colonization in ventilated preterm infants.

The role of Acinetobacter baumannii in infections in ventilated preterm infants was evaluated in 15 colonized infants (11 male, 4 female) in a pediatric intensive care unit. These cases were randomly matched by birth weight and gestational age with ventilated non-colonized controls (8 male, 7 female). Case records were reviewed for signs and symptoms of infection. Colonized infants were ventilated significantly longer (p < 0.05) than controls, and had body temperatures of > 37 degrees C for a significantly longer period of time (p < 0.05). No other parameter of infection differed significantly between the groups. The duration of intensive care treatment did not differ between cases and controls, nor did the weight gain during intensive care treatment. No fatalities occurred in either group.

Acinetobacter↗

Clinical and bacteriological features of relapsing shunt-associated meningitis due to Acinetobacter baumannii.

A case is reported of a patient who developed Acinetobacter baumannii meningitis ten days after a ventriculoperitoneal shunt had been placed for control of elevated intracranial pressure. Intravenous antimicrobial therapy was instituted with imipenem and tobramycin after shunt removal and insertion of a ventriculostomy tube. Following the placement of a ventriculoatrial shunt, clinical and microbiological relapse occurred that was eventually cured after complete removal of the ventricular drainage system and a second course of systemic antibiotics. Relapse was confirmed by modern molecular typing techniques including plasmid DNA fingerprinting and analysis of total genomic DNA by pulsed-field gel electrophoresis.

Acinetobacter↗

Characteristics of clinical Acinetobacter spp. strains.

Resistance to 13 antimicrobial agents, resistance to the bactericidal activity of human serum, hydrophobic properties, lipolytic activity and production of histamine were determined in a total of 50 clinical Acinetobacter spp. strains (A. baumannii, A. lwoffii, A. calcoaceticus, A. haemolyticus). None of the tested isolates showed resistance to meropenem and none of A. lwoffii, A. calcoaceticus and A. haemolyticus strains were resistant to amikacin. Forty-six strains (92%) manifested resistance to ampicillin, 90% to cefuroxime, 68% to ciprofloxacin, 58% to piperacillin, gentamicin and cotrimaxazole, 50% to cefotaxime, 44% to amikacin, 42% to ceftazidime, 38% to piperacillin/tazobactam, 24% to netilmicin and 16% to ampicillin/sulbactam. In particular, A. baumannii and A. calcoaceticus strains showed considerable antibiotic resistance. Thirty-one isolates (62%) showed serum resistance; intermediate sensitivity was found in 19 isolates (38%). The majority of the strains (72%) demonstrated a strongly hydrophobic character; 16% of isolates exhibited moderate hydrophobic properties. All strains showed lipolytic activity; production of histamine was detected in 14 of 43 strains examined.

Acinetobacter↗

Tetracyclines for treating multidrug-resistant Acinetobacter baumannii ventilator-associated pneumonia.

OBJECTIVE: To report the use of tetracyclines for the treatment of multidrug-resistant Acinetobacter baumannii ventilator-associated pneumonia (VAP). DESIGN: Observational case series. SETTING: . The Presley Regional Trauma Center located within the Regional Medical Center, Memphis, Tennessee, USA. PATIENTS AND PARTICIPANTS: Seven critically ill trauma patients with VAP caused by A. baumannii isolates that were resistant to all antibiotics tested except for doxycycline or minocycline. INTERVENTIONS: Patients were treated with IV doxycycline or minocycline for an average of 13.5 (range 9-20) days. MEASUREMENTS AND RESULTS: Doxycycline or minocycline was successful in six of seven patients. CONCLUSIONS: Doxycycline or minocycline may be effective for treating multidrug-resistant A. baumannii VAP.

APACHE↗

Acinetobacter baumannii ventilator-associated pneumonia: epidemiological and clinical findings.

OBJECTIVE: To investigate prognostic factors and predictors of Acinetobacter baumannii isolation in ventilator-associated pneumonia (VAP). We specifically analyzed these issues for imipenem-resistant episodes. DESIGN AND SETTING: All episodes of VAP are prospectively included in a database. Information about risk factors was retrieved retrospectively. PATIENTS: Eighty-one patients exhibiting microbiologically documented VAP: 41 by A. baumannii (26 by imipenem-resistant) and 40 by other pathogens. MEASUREMENTS AND RESULTS: The following variables were noted: underlying diseases, severity of illness, duration of mechanical ventilation and of hospitalization before VAP, prior episode of sepsis, previous antibiotic, corticosteroid use, type of nutrition, renal replacement therapy, reintubation, transportation out of the ICU, micro-organisms involved in VAP, concomitant bacteremia, clinical presentation, Sequential Organ Failure Assessment (SOFA) scale on the day of diagnosis, and adequacy of empirical antibiotic therapy. Prior antibiotic use was found to be associated with development of VAP by A. baumannii (OR 14). Prior imipenem exposure was associated with the isolation of imipenem-resistant strains (OR 4). SOFA score on the day of diagnosis was the only predictor of in-hospital mortality (OR 1.22); adequacy of empirical antibiotic therapy was a protective factor (OR 0.067). CONCLUSIONS: Our results confirm that prior exposure to antimicrobials is an independent predictor for the development of A. baumannii VAP, the prognosis of which is similar to that of infections caused by other pathogens. This study highlights the importance of initial antibiotic choice in VAP or whatever cause.

Acinetobacter Infections↗

Acquisition of imipenem-resistant Acinetobacter baumannii in a pediatric intensive care unit: A case-control study.

OBJECTIVE: To investigate the risk factors associated with nosocomial acquisition of imipenem-resistant Acinetobacter baumannii (IRAB) among pediatric intensive care patients. A retrospective case control study was conducted in a pediatric intensive care unit (PICU). PATIENTS AND PARTICIPANTS: Cases were children in whom IRAB was isolated from any clinical specimen obtained at least 48 h following admission to PICU. Controls were children without IRAB matched to cases in 2:1 ratio. Twenty-six cases were matched with 52 controls according to the chronological order of admission. MEASUREMENTS AND RESULTS: Between July 2001 and December 2003, 52 (62%) of 84 clinical A. baumannii isolates were found nonsusceptible to imipenem (MIC > or = 8 microg/ml). Demographic variables, comorbid conditions, clinical picture at admission, invasive procedures, use of antimicrobials and other drugs were analyzed as potential risk factors. Use of carbapenems and other beta-lactams, aminoglycosides, ranitidine, mechanical ventilation, central venous or urinary catheters and length of stay in PICU were among the factors significantly associated with IRAB acquisition in the univariate analysis. By multivariate analysis, however, only aminoglycoside use and length of stay in the PICU were independent risk factors. CONCLUSIONS: Acquisition of IRAB by PICU patients was independently associated with aminoglycoside use and prolonged stay in the unit. Studies of evaluation of infection control policies need to be pursued.

Acinetobacter Infections↗

Antibiotic-resistant Acinetobacter baumannii can be killed by a combination of bacteriophages and complement.

Infections caused by multidrug-resistant Acinetobacter baumannii are an emerging global health threat. Although phages have shown promising results in treating bacterial infections, the mechanisms of the combined effect of phages and innate immunity on clearing A. baumannii remain unclear. Here, we report a synergistic effect of the complement system and phages on clearing multidrug-resistant A. baumannii. We show that A. baumannii rapidly adapts and becomes resistant to phage or serum complement by modifying the expression of capsule and lipooligosaccharides, which can be regulated through reversible transposon mutagenesis in the K locus. Compared to the encapsulated phenotype, the non-encapsulated, phage-resistant A. baumannii showed a higher level of membrane attack complex deposition and were susceptible to killing by complement. In contrast, the encapsulated phenotype escaped the complement system by shedding the membrane attack complex to the environment. Thus, while the complement system targets the non-encapsulated phenotype, the phage infects and eliminates the encapsulated subpopulation. These results suggest means of combatting antibiotic-resistant A. baumannii by a simultaneous treatment with phages and complement, a combination which can be supplemented further with antibacterial antibodies.

Acinetobacter baumannii↗

Cerebrospinal fluid penetration and pharmacokinetic/pharmacodynamic parameters of intravenously administered colistin in a case of multidrug-resistant Acinetobacter baumannii meningitis.

Described here is a case of meningitis caused by multidrug-resistant Acinetobacter baumannii susceptible only to colistin, which was treated successfully with intravenous colistin sulfomethate sodium (5 mg/kg/day). The levels of colistin in serum and cerebrospinal fluid and the pharmacokinetic/pharmacodynamic parameters of colistin were determined. In this case, intravenously administered colistin penetrated cerebrospinal fluid (25% of serum levels) at levels sustaining bactericidal concentrations.

Acinetobacter↗

Successful treatment of septic shock due to pan-resistant Acinetobacter baumannii using combined antimicrobial therapy including tigecycline.

Reported here is the case of a patient with septic shock due to multidrug-resistant Acinetobacter baumannii, which developed after complicated acute pancreatitis with intra-abdominal abscess. Treatment with colistin methanesulphonate and high doses of meropenem were initiated, but since shock persisted, tigecycline was added to the regimen, resulting in successful resolution of the infection.

Acinetobacter Infections↗

Analysis of prognostic factors in 95 patients with Acinetobacter baumannii bacteremia.

BACKGROUND: Because Acinetobacter baumannii bacteremia is a global problem, we were motivated to characterize this disease in Taiwan. PATIENTS AND METHODS: We analyzed findings in 95 patients with documented A. baumannii bacteremia between January 1, 1998 and December 31, 2000 (47 men, 48 women; mean age 58.8 years). RESULTS: The mean length of stay in the hospital was 44.0 days. Clinically, 76 patients had fever and 35 patients developed shock. Fifty patients had respiratory tract infections; 24, urinary tract infections; 11, intra-abdominal infections; three, CNS infections; and two, catheter-related infections. Five patients had primary bacteremia. Empirical antibiotic therapy was initiated at the onset of the clinical signs of infection. Antimicrobial susceptibility test results were variable. 47 patients died and 48 survived; the mortality rate for A. baumannii bacteremia was 45.3% (43/95). CONCLUSION: Physicians should pay attention to this infection because the early identification of high-risk patients could facilitate prophylaxis and potentially reduce associated problems.

Acinetobacter Infections↗

Nosocomial respiratory tract infection and colonization with Acinetobacter calcoaceticus. Epidemiologic characteristics.

Nosocomial respiratory tract infection with Acinetobacter calcoaceticus occurs frequently in many hospitals. An outbreak of respiratory tract infections in an intensive care unit provided an opportunity to study clinical and epidemiologic characteristics of such infections. Retrospective studies demonstrated that A. calcoaceticus in sputum was significantly associated with endotracheal intubation (p = 0.03) and continuous positive pressure ventilation (p less than 0.02). After control measures had interrupted the outbreak, a prospective microbiologic investigation demonstrated that one third of the hospital personnel had transient hand colonization with multiple strains of A. calcoaceticus. Pharyngeal, vaginal and rectal carriage was rare. A pulmonary therapist with chronic dermatitis had persistent hand colonization with the epidemic strain, and he contaminated respiratory therapy equipment. Cross contamination of respiratory therapy equipment occurred while in use, but no other inanimate reservoir was demonstrated. Although previous studies have implied that the inanimate hospital environment has unique reservoirs of A. calcoaceticus, these reservoirs were not implicated in this outbreak. Human skin must be considered an important reservoir of A. calcoaceticus.

Acinetobacter↗