Indian J Med Res, 2004 Dec, 120, 553 - 556 Evaluation of extended spectrum beta lactamase in urinary isolates; Tankhiwale SS et al.; BACKGROUND & OBJECTIVES: Urinary tract infection (UTI) remain the common infections diagnosed in outpatients as well as in hospitalized patients . Current knowledge on antimicrobial susceptibility pattern of uropathogens is mandatory for appropriate therapy . Extended spectrum beta lactamases (ESBL) hydrolyse expanded spectrum cephalosporins like ceftazidime, cephotaxime which are used in the treatment of UTI . ESBL producing bacteria may not be detectable by routine disk diffusion susceptibility test, leading to inappropriate use of antibiotics and treatment failure . Not much information on ESBL producing organisms causing UTI is available from India . An effort was therefore made to study the ESBL producing uropathogens and also the susceptibility patterns of ESBL and nonESBL producers . METHODS: Urinary isolates from symptomatic UTI cases attending or admitted to the Indira Gandhi Medical College and Hospital, Nagpur were identified by conventionl methods . Antimicrobial susceptibility testing was done by Kirbey Bauer's disc diffusion method . Isolates resistant to cephotaxime were tested for ESBL production by double disc synergy test method . RESULTS: Of the 217 isolates, 87 were cephotaxime resistant Gram-negative bacilli . Of these, 42 (48.3%) were found to be ESBL producers . Escherichia coli, Klebsiella pnuemoniae and Acinetobacter were ESBL producing species . Multidrug resistance was found to be significantly (P<0.05) more in ESBL producing isolates (90.5%) than non ESBL producers (68.9%) . INTERPRETATION & CONCLUSION: In the present study a large number of uropathogens were found to be ESBL producers . Most of the ESBL producing isolates were multidrug resistant . Monitoring of ESBL production and antimicrobial susceptibility testing are necessary to avoid treatment failure in patients with UTI. Microb Drug Resist, 2004 Winter, 10(4), 292 - 9 Variable Resistance Patterns of Integron-Associated Multidrug-Resistant Acinetobacter baumannii Isolates in a Surgical Intensive Care Unit; Wu TL et al.; Between 1998 and 2000, we characterized 91 nosocomial isolates of Acinetobacter baumannii by antibiotyping and genotyping . A total of 25 ribotypes were obtained among these 91 isolates . When the isolates from surgical intensive care units (ICUs) and other wards were compared, multiresistant A . baumannii isolates with the same ribotype 25 (R-25) were significantly more prevalent in nosocomial infections among the surgical patients . Further subtyping of the strains by pulsed-field gel electrophoresis confirmed that strains of the same ribotype in surgical ICUs and a few isolates from other wards were identical or clonally related . Different antibiotic resistance profiles were observed among these R-25 isolates . All R-25 isolates contained intI1 integrase, and two clusters of integron cassettes were found . These clusters of cassettes were encoded by an open reading frame (ORF) of -5'CS-aac(3)Ia-aadA1a-unknown or f-3'CS or 5'CS-aacA4-aadA1-catB8-3'CS, indicating the involvement of different resistant genes . Two isolates contained bla (IMP-1), which was acquired from a conjugatively transferable plasmid and did not involve integron-associated resistance . In conclusion, an epidemic of nosocomial infections associated with A . baumannii strains that have different resistance profiles was identified . Resistance profiles can change by a combination of plasmid- and integron-associated acquisition, especially in a unit with high antibiotic selective pressures . Infectious control personnel should be alert to the change in resistance profiles during routine monitoring. Microb Drug Resist, 2004 Winter, 10(4), 286 - 91 Genotypic Characterization of Carbapenem-Nonsusceptible Acinetobacter spp . Isolated in Latin America; Gales AC et al.; The principal aim of this study was to evaluate the genomic diversity among the imipenem-nonsusceptible Acinetobacter spp . (INSA) collected from the Latin American medical centers within the SENTRY Antimicrobial Surveillance Program . The INSA isolates were collected from patients with bloodstream infections, who were hospitalized in seven Latin American countries between 1997 and 1999 . For epidemiologic comparison, 20 carbapenem-susceptible Acinetobacter spp . (CSA) isolates were collected in the same period of time from the respective medical centers . A total of 23 Acinetobacter spp . isolates exhibiting imipenem MIC values of >/=8 microg/ml were typed by ribotyping, an automated molecular method . The isolates showing an identical ribogroup were also typed by pulsed-field gel electrophoresis (PFGE) . The antimicrobial susceptibility to various antimicrobial agents was evaluated using a reference broth microdilution technique . Among the INSA isolates, 13 distinct ribogroups were observed, whereas 16 ribogroups were detected among the CSA . Nearly 57% of the INSA belonged to only four ribogroups . Identical ribogroups and PFGE patterns were observed among INSA and CSA isolates collected from medical centers located in different countries (Brazil and Argentina) . Our results showed: (1) a higher genomic variability among the CSA; (2) presence of epidemic clones among INSA isolates encountered in Latin American medical centers; and (3) spread of INSA and CSA epidemic clones between Latin American countries. Clin Microbiol Infect, 2005 Jan, 11(1), 24 - 30 Activities of various beta-lactams and beta-lactam/beta-lactamase inhibitor combinations against Acinetobacter baumannii and Acinetobacter DNA group 3 strains; Brauers J et al.; Acinetobacter baumannii and Acinetobacter DNA group 3 are members of the so-called A . calcoaceticus-A . baumannii complex and are important nosocomial pathogens . Multiresistance in these organisms is increasingly frequent, and alternative treatment options are needed . The beta-lactamase inhibitors clavulanate, sulbactam and tazobactam have intrinsic activity against Acinetobacter strains . In the present study, broth microdilution was used to assess the in-vitro activities of currently available beta-lactam/beta-lactamase inhibitor combinations and sulbactam alone against 469 Acinetobacter isolates (A . baumannii, n = 395; Acinetobacter DNA group 3, n = 74) collected from various laboratories in Germany . Fixed concentrations and fixed ratios of beta-lactamase inhibitors were used . Sulbactam-containing combinations (susceptibility rates of 90.4-92.7% for A . baumannii and 97.3-100% for Acinetobacter DNA group 3) and sulbactam alone were superior to clavulanate- and tazobactam-containing combinations . The activity of sulbactam-containing combinations against members of the A . calcoaceticus-A . baumannii complex was conferred exclusively by the intrinsic activity of the beta-lactamase inhibitor and did not result from enhanced beta-lactam activity . Testing with the inhibitor added at a fixed ratio of inhibitor to beta-lactam appeared to give more reliable results than testing at a fixed concentration of the inhibitor . Resistance to carbapenems (0.3%) remains low in Germany. Clin Microbiol Infect, 2005 Jan, 11(1), 15 - 23 Characterisation of OXA-51, a novel class D carbapenemase found in genetically unrelated clinical strains of Acinetobacter baumannii from Argentina; Brown S et al.; Acinetobacter baumannii is now one of the most frequently encountered nosocomial pathogens in intensive therapy units, and is renowned for being difficult to treat because of resistance to most antibiotics . Carbapenems are the remaining drugs of choice in many centres, but carbapenem resistance is now emerging in strains worldwide . Two subgroups of carbapenem-hydrolysing beta-lactamases, which differ in their amino-acid homology, have been found in some resistant strains . This report describes the emergence and characterisation of a novel carbapenemase (OXA-51) in genetically distinct carbapenem-resistant A . baumannii strains from Argentina . Enzyme kinetics and inhibitor studies were performed spectrophotometrically with purified beta-lactamase . Amplification of the gene was achieved with a two-step PCR method employing arbitrary partially degenerate and gene-specific primers . Transfer of imipenem resistance was attempted with the use of broth and membrane filter methods . Attempts to produce plasmid-cured variants were made in ethidium bromide curing experiments . OXA-51 was identified in two clones of A . baumannii, and was found to have < 63% amino-acid identity with subgroups 1 and 2 . Enzyme kinetic studies confirmed that OXA-51 was a molecular class D enzyme with carbapenemase activity, and that it displayed the highest affinity for imipenem (K(m) value 11 microM) . Sequence analysis of the gene identified distinct differences within conserved class D motifs when compared with subgroups 1 and 2 . Attempts to transfer imipenem resistance and to determine a plasmid location for the gene failed . OXA-51 is the first of a new subgroup of carbapenemases to emerge in multiresistant clinical isolates of A . baumannii. Can J Microbiol, 2004 Nov, 50(11), 935 - 41 Degradation of acyl-homoserine lactone molecules by Acinetobacter sp . strain C1010; Kang BR et al.; A bacterium C1010, isolated from the rhizospheres of cucumbers in fields in Korea, degraded the microbial quorum-sensing molecules, hexanoyl homoserine lactone (HHSL), and octadecanoyl homoserine lactone (OHSL) . Morphological characteristics and 16S rRNA sequence analysis identified C1010 as Acinetobacter sp . strain C1010 . This strain was able to degrade the acyl-homoserine lactones (AHLs) produced by the biocontrol bacterium, Pseudomonas chlororaphis O6, and a phytopathogenic bacterium, Burkholderia glumae . Co-cultivation studies showed that the inactivation of AHLs by C1010 inhibited production of phenazines by P . chlororaphis O6 . In virulence tests, the C1010 strain attenuated soft rot symptom caused by Erwinia carotovora ssp . carotovora . We suggest Acinetobacter sp . strain C1010 could be a useful bacterium to manipulate biological functions that are regulated by AHLs in various Gram-negative bacteria. Arch Neurol, 2005 Jan, 62(1), 33 - 36 Acinetobacter Immune Responses in Multiple Sclerosis: Etiopathogenetic Role and Its Possible Use as a Diagnostic Marker; Ebringer A et al.; Multiple sclerosis (MS) is the most common cause of neurologic disability among young people . The etiology of MS is controversial, but immune responses are considered to somehow be involved . The diagnosis of MS depends on a combination of various clinical and laboratory features, but apart from some myelin-neuronal autoantibody profiles or oligoclonal bands in the cerebrospinal fluid no other serologic diagnostic test or marker has yet been discovered . However, the presence of antibodies to Acinetobacter species in MS patients opens the possibility of developing a composite laboratory diagnostic marker, the myelin-Acinetobacter-neurofilament index . Whether Acinetobacter is the triggering agent of MS remains to be determined, but the measurement of anti-Acinetobacter antibodies could be used as a marker of disease activity . To evaluate this, prospective randomized controlled studies should be performed with MS patients, especially in the early stages of the disease. BMC Infect Dis . 2005 Jan 10;5(1):1 {Epub ahead of print} Toxicity after prolonged (more than four weeks) administration of intravenous colistin; Falagas ME et al.; BACKGROUND: The intravenous use of polymyxins has been considered to be associated with considerable nephrotoxicity and neurotoxicity . For this reason, the systemic administration of polymyxins had been abandoned for about 20 years in most areas of the world . However, the problem of infections due to multidrug-resistant (MDR) Gram-negative bacteria such us Pseudomonas aeruginosa and Acinetobacter baumanniii has led to the re-use of polymyxins . Our objective was to study the toxicity of prolonged intravenous administration of colistin (polymyxin E) . METHODS: An observational study of a retrospective cohort at "Henry Dunant" Hospital, a 450-bed tertiary care center in Athens, Greece, was undertaken . Patients who received intravenous colistin for more than 4 weeks for the treatment of multidrug resistant Gram-negative infections were included in the study . Serum creatinine, blood urea, liver function tests, symptoms and signs of neurotoxicity were the main outcomes studied . RESULTS: We analyzed data for 19 courses of prolonged intravenous colistin {mean duration of administration (+/- SD) 43.4 (+/- 14.6) days, mean daily dosage (+/- SD) 4.4 (+/- 2.1) million IU, mean cumulative dosage (+/- SD) 190.4 (+/- 91.0) million IU} in 17 patients . The median creatinine value increased by 0.25 mg/dl during the treatment compared to the baseline (p< 0.001) but returned close to the baseline at the end of treatment (higher by 0.1 mg/dl, p= 0.67) . No apnea or other evidence of neuromuscular blockade was noted in any of these patients who received prolonged treatment with colistin . CONCLUSIONS: No serious toxicity was observed in this group of patients who received prolonged intravenous colistin . Colistin should be considered as a therapeutic option in patients with infections due to multidrug resistant Gram-negative bacteria. Srp Arh Celok Lek, 2001 May-Jun, 129 Suppl 1, 36 - 41 {Bacterial causes of meningitis in newborns} {PCR genotyping of Acinetobacter hospital isolates} {No authors listed} A total of 13 Acinetobacter baumannii-13TU isolates obtained from patients of the Perm regional clinical hospital during the period of 1 year, were genotyped in the polymerase chain reaction (PCR) with universal primers . Acinetobacter cultures could not be distinguished by phenotypic tests and antibiogram and were resistant to B-lactams and gentamicin . According to the results of amplification the obtained isolates could be subdivided into two groups (with 6 and 7 respectively) . The detection of "epidemic" strains, including those capable of prolonged (for more than 5 months) persistence in hospital environment, may be indicative of the growing role of Acinetobacter as the causative agent in nosocomial infection. Appl Microbiol Biotechnol, 2005 Feb, 66(5), 536 - 41 Epub 2004 Aug 04. The AlnB protein of the bioemulsan alasan is a peroxiredoxin; Bekerman R et al.; The bioemulsifier of Acinetobacter radioresistens KA53, referred to as alasan, is a high molecular weight complex of a polysaccharide and three proteins (AlnA, AlnB and AlnC) . AlnA has previously been shown to be an OmpA-like protein that is largely responsible for the emulsifying activity of alasan . To further elucidate the nature of alasan, the gene coding for AlnB was cloned, sequenced and overexpressed in Escherichia coli . The overall 561 bp sequence of the hypothetical AlnB showed strong homology, including all conserved regions and residues known to be essential for enzymatic activity, to the ubiquitous family of thiol-specific antioxidant enzymes known as peroxiredoxins . Transgenic E . coli overexpressing AlnB exhibited increased resistance to cumene hydroperoxide both in liquid culture and on agar medium . Recombinant AlnB had no emulsifying activity but stabilized oil-in-water emulsion generated by AlnA. Theriogenology, 2005 Jan 15, 63(2), 573 - 84 Bacteriospermia in extended porcine semen; Althouse GC et al.; Bacteriospermia is a frequent finding in freshly extended porcine semen and can result in detrimental effects on semen quality and longevity if left uncontrolled . The primary source of bacterial contamination is the boar . Other sources that have been identified include environment, personnel, and the water used for extender preparation . A 1-year retrospective study was performed on submissions of extended porcine semen for routine quality control bacteriological screening at the University of Pennsylvania . Out of 250 sample submissions, 78 (31.2%) tested positive for bacterial contamination . The most popular contaminants included Enterococcus spp . (20.5%), Stenotrophomonas maltophilia (15.4%), Alcaligenes xylosoxidans (10.3%), Serratia marcescens (10.3%), Acinetobacter lwoffi (7.7%), Escherichia coli (6.4%), Pseudomonas spp . (6.4%), and others (23.0%) . Prudent individual hygiene, good overall sanitation, and regular monitoring can contribute greatly in controlling bacterial load . Strategies that incorporate temperature-dependent bacterial growth and hyperthermic augmentation of antimicrobial activity are valuable for effective control of susceptible bacterial loads . Aminoglycosides remain the most popular antimicrobial class used in porcine semen extenders, with beta-lactam and lincosamide use increasing . With the advent of more novel antimicrobial selection and semen extender compositions in swine, prudent application and understanding of in vitro pharmacodynamics are becoming paramount to industry success in the use of this breeding modality. Jpn J Infect Dis, 2004 Dec, 57(6), 273 - 5 Antimicrobial susceptibility of isolates from neonatal septicemia; Agnihotri N et al.; A retrospective study of bacterial isolates from cases of neonatal septicemia was undertaken over a period of 5 years (July 1998 - June 2003) at the Government Medical College Hospital, Chandigarh, India . The study was carried out to determine the bacterial profile, the antimicrobial susceptibility of the isolates, and the change in trends over the study period . A total of 3,064 blood samples for blood culture were obtained, out of which 588 were positive for bacterial isolates . Most of the cases detected by blood culture occurred in the first week of life (64.4%) . Gram-negative bacilli (58.5%) predominated over Gram-positive cocci (41.5%) . Staphylococcus aureus was found to be the most common isolate (35.0%).The incidence of Gram-positive and Gram-negative organisms changed little over the 5 year span . However, a constant and significant rise in the incidence of Acinetobacter spp . was observed between the first to fifth year of the study period . Amikacin was found to be the most effective drug against Gram-negative bacteria . For S . aureus and Pseudomonas aeruginosa, overall resistance percentages of 5 years show that netilmicin and ciprofloxacin, respectively, were the most effective drugs. Int J Antimicrob Agents, 2005 Jan, 25(1), 57 - 61 Dissemination and diversity of metallo-beta-lactamases in Latin America: report from the SENTRY Antimicrobial Surveillance Program; Sader HS et al.; Carbapenem resistance among Pseudomonas aeruginosa and Acinetobacter spp . is becoming a critical therapeutic problem worldwide . The SENTRY Antimicrobial Surveillance Program monitors pathogen frequency and antimicrobial resistance patterns of nosocomial and community-acquired infections through sentinel hospitals on five continents . Pseudomonas spp . and Acinetobacter spp . strains resistant to imipenem (MIC, >/=16mg/l), meropenem (MIC, >/=16mg/l), and ceftazidime (MIC, >/=32mg/l) collected from January 2001 to December 2003 were routinely screened for antimicrobial resistance genes . Resistant isolates were initially tested for metallo-beta-lactamase (MbetaL) production by phenotypic tests (disk approximation or MbetaL Etest strip) and then characterization of the MbetaL (hydrolysis assays, PCR for bla(IMP), bla(VIM), bla(SPM), gene sequencing) . Eighty-nine isolates (33 Acinetobacter spp., 54 Pseudomonas aeruginosa, and 2 P . fluorescens) had positive phenotypic screening tests . Among those, 34 isolates producing MbetaL were identified, including 7 Acinetobacter spp., 25 P . aeruginosa and 2 P . fluorescens . The MbetaLs identified were IMP-1, VIM-2 and two newly described enzymes: SPM-1 and IMP-16 . The greatest concentration of MbetaL strains was in Brazil, where imipenem-resistant P . aeruginosa increased significantly in the time period evaluated by the SENTRY Program . MbetaL-producing P . aeruginosa was detected in Sao Paulo (SPM-1) and Brasilia (SPM-1 and IMP-16), Brazil and Caracas, Venezuela (VIM-2); while MbetaL-producing Acinetobacter spp . isolates were detected in Sao Paulo, Brazil (IMP-1) . P . fluorescens isolates producing IMP-1 and VIM-2 were detected in Sao Paulo, Brazil and Santiago, Chile, respectively . The emergence and dissemination of mobile MbetaL-producing isolates represent an alarming factor for increasing resistance to carbapenems in several medical centres evaluated by the SENTRY Program in Latin America. Int J Antimicrob Agents, 2005 Jan, 25(1), 11 - 25 Evaluation of colistin as an agent against multi-resistant Gram-negative bacteria; Li J et al.; Infections caused by multi-resistant Gram-negative bacteria, particularly Pseudomonas aeruginosa, are increasing worldwide . In patients with cystic fibrosis (CF), resistance in P . aeruginosa to numerous anti-pseudomonal agents is becoming common . The absence since 1995, of new substances active against resistant Gram-negative bacteria, has caused increasing concern . Colistin, an old antibiotic also known as polymyxin E, has attracted more interest recently because of its significant activity against multi-resistant P . aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae, and the low resistance rates to it . Because its use as an anti-pseudomonal agent was displaced by the potentially less toxic aminoglycosides in 1970s, our knowledge of this drug is limited . However, there has been a significant recent increase in the data gathered on colistin, focussing on its chemistry, antibacterial activity, mechanism of action and resistance, pharmacokinetics, pharmacodynamics and new clinical application . It is likely that colistin will be an important antimicrobial option against multi-resistant Gram-negative bacteria, for some years to come. J Hosp Infect, 2005 Feb, 59(2), 113 - 25 Assessment of the antibacterial activity of tea tree oil using the European EN 1276 and EN 12054 standard suspension tests; Messager S et al.; The activity of tea tree oil (TTO) and TTO-containing products was investigated according to the EN 1276 and EN 12054 European suspension methods . The activity of different concentrations of TTO, a hygienic skin wash (HSW), an alcoholic hygienic skin wash (AHSW) and an alcoholic hand rub (AHR) was investigated . These formulations were assessed in perfect conditions with the EN 12054 test, and in perfect conditions as well as in the presence of interfering substances with the EN 1276 test, against Staphylococcus aureus, Acinetobacter baumannii, Escherichia coli and Pseudomonas aeruginosa . With the latter test, the activity of the same formulations without TTO was also assessed as a control . With the EN 1276 test, the AHR achieved a >10(5)-fold reduction against all four test organisms within a 1-min contact time . The AHSW achieved a >/=10(5)-fold reduction against A . baumannii after a 1-min contact time and against S . aureus, E . coli and P . aeruginosa after a 5-min contact time . The efficacy of TTO appeared to be dependent on the formulation and the concentration tested, the concentration of interfering substances and, lastly, the organism tested . Nevertheless, 5% TTO achieved a >10(4)-fold reduction in P . aeruginosa cell numbers after a 5-min contact time in perfect conditions . TTO (5%) in 0.001% Tween 80 was significantly more active against E . coli and P . aeruginosa than against S . aureus and A . baumannii . With the EN 12054 test, after a 1-min contact time, 5% TTO in 0.001% Tween 80 and the AHSW achieved a >10(4)-fold reduction in E . coli and A . baumannii cell numbers, respectively, and the AHR achieved a >4log(10) reduction against all organisms tested . The formulations used in this study are now being tested using a novel ex vivo method as well as the in vivo European standard handwashing method EN 1499. Med Hypotheses, 2005, 64(3), 487 - 494 A possible link between multiple sclerosis and Creutzfeldt-Jakob disease based on clinical, genetic, pathological and immunological evidence involving Acinetobacter bacteria; Ebringer A et al.; Multiple sclerosis (MS) is an immune-mediated demyelinating disease of the nervous system . There is an increasingly likelihood that MS could be triggered/perpetuated by environmental (microbial) agents . Sporadic Creutzfeldt-Jakob disease (sCJD) is a relatively rare but fatal disease, which shows various clinical, genetic, pathological and immunological features through which it resembles a severe form of MS . The disease in some patients with MS may show a rapidly downhill course with death occurring within one to two years and a similar situation occurs in sCJD . The occurrence of these comparative similarities between MS and sCJD could be explained on the basis that both of these conditions might be sharing a common aetiopathogenic factor such as infection by Acinetobacter microbes and this possibility could be investigated further by carrying out immunological studies on a relatively large numbers of patients with MS and CJD. Antimicrob Agents Chemother, 2005 Jan, 49(1), 202 - 8 OXA-58, a novel class D {beta}-lactamase involved in resistance to carbapenems in Acinetobacter baumannii; Poirel L et al.; A carbapenem-resistant Acinetobacter baumannii strain was isolated in Toulouse, France, in 2003 . Cloning and expression in Escherichia coli identified the carbapenem-hydrolyzing beta-lactamase OXA-58, which is weakly related (less than 50% amino acid identity) to other oxacillinases . It hydrolyzed penicillins, oxacillin, and imipenem but not expanded-spectrum cephalosporins . The bla(OXA-58) gene was located on a ca . 30-kb non-self-transferable plasmid . After electrotransformation in the A . baumannii CIP7010(T) reference strain, it conferred reduced susceptibility to carbapenems . The bla(OXA-58) gene was bracketed by two novel ISAba3-like insertion elements . This study describes a newly characterized beta-lactamase that may contribute to carbapenem resistance in A . baumannii. Srp Arh Celok Lek, 2004 Oct, 132 Suppl 1, 49 - 53 {Initial antibiotic therapy of neonatal sepsis}; Emulsan quantitation by Nile red quenching fluorescence assay; Department of Biomedical Engineering, School of Engineering, Tufts University, 4 Colby Street, Medford, MA, 02155, USA, david.kaplan@tufts.eduA Nile red fluorescent technique to quantify 20-200 mug ml(-1) of emulsan was developed . Nile red dissolved in DMSO showed an adsorption peak at 552 nm, and emission peak at 636 nm, with molar extinction coefficient of 19,600 cm(-1) M(-1) . Nile red fluorescence in DMSO was proportionally quenched by emulsan and the quenching was time-dependent . The assay was used to follow the production of emulsan by cultures of Acinetobacter venetianus RAG-1. JAMA, 2004 Dec 22, 292(24), 3006 - 11 An outbreak of multidrug-resistant Acinetobacter baumannii associated with pulsatile lavage wound treatment; Maragakis LL et al.; 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. Clin Microbiol Infect, 2004 Dec, 10(12), 1098 - 101 Synergic in-vitro activity of imipenem and sulbactam against Acinetobacter baumannii; Choi JY et al.; 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. Biotechnol Lett, 2004 Nov, 26(21), 1639 - 42 Isolation and characterization of Acinetobacter sp . ES-1 excreting a lipase with high enantioselectivity for ( S )-ketoprofen ethyl ester; Lee KW et al.; A new Acinetobacter sp . ES-1, grown on triolein, tryptone and Triton X-100, excreted a lipase that hydrolyzed 10m M ( R , S )-ketoprofen ethyl ester into ( S )-ketoprofen . The crude lipase had an activity of 10Uml(-1) and, at 30 degrees C and pH7 over 48h, gave a conversion yield of 35% with an enantiomeric excess for the product 96%. Int J Infect Dis, 2005 Jan, 9(1), 43 - 51 Clinical and microbiologic determinants of serious bloodstream infections in Egyptian pediatric cancer patients: a one-year study; El-Mahallawy H et al.; OBJECTIVES:: Bloodstream infections (BSI) remain a major cause of morbidity and death in patients undergoing treatment for cancer . However, all recent epidemiological and therapeutic studies underline the absolute need for knowledge of the factors governing the infections in each center . The aim of this study is to identify the factors affecting BSI in the pediatric service of the National Cancer Institute (NCI) at Cairo University . More tailored policies for the treatment of patients with febrile neutropenia following chemotherapy can then be created . PATIENTS AND METHODS:: Over a 12-month period, all children with cancer and fever, with or without neutropenia, who were admitted to the NCI for empirical therapy of febrile episodes and who had a microbiologically confirmed bloodstream infection were studied retrospectively . RESULTS:: A total of 328 BSI occurred in 1135 febrile episodes in pediatric cancer patients at the NCI in one year . Gram-positive bacteria were isolated in 168 episodes (51.2%) and 61.9% of the total isolates (either single or mixed), Gram-negative in 97 (29.6%), and mixed infections in 45 (13.7%) . The common causative agents of bloodstream infections in this study were coagulase-negative staphylococci (16.2%), Staphylococcus aureus (13.4%), Streptococcus spp . (12.1%) followed by Acinetobacter spp . (6.7%) and Pseudomonas spp . (5.5%) . Fungemia was encountered in 18 episodes, being mixed in nine of them . A more serious BSI in terms of a prolonged episode was encountered in 30.2% of the episodes and was significantly associated with patients being hospitalized, having intensified chemotherapy, polymicrobial and fungal infection, lower respiratory tract infections and persistent neutropenia at day seven . CONCLUSIONS:: In a large population of children, common clinical and laboratory risk factors were identified that can help predict more serious BSI . These results encourage the possibility of a more selective management strategy for these children. FEMS Microbiol Lett, 2004 Dec 15, 241(2), 157 - 62 Quinate oxidation in Gluconobacter oxydans IFO3244: purification and characterization of quinoprotein quinate dehydrogenase; Vangnai AS et al.; Quinoprotein quinate dehydrogenase (QDH) is a membrane-bound enzyme containing pyrroloquinoline quinone (PQQ) as the prosthetic group . QDH in Gluconobacter oxydans IFO3244 was found to be inducible by quinate and it is not constitutively expressed in the absence of quinate . The purification of holo-form of QDH to nearly homogeneity was achieved . The purified QDH appears to have two subunits of approximately 65 and 21 kDa, which could be the result of proteolysis of single polypeptide . Kinetic analysis indicated that the purified enzyme is much more specific to quinate than QDH from Acinetobacter calcoaceticus . The efficiency of the artificial electron acceptor was also determined. J Antimicrob Chemother, 2005 Jan, 55(1), 115 - 118 Epub 2004 Dec 8. A nosocomial outbreak of Acinetobacter baumannii isolates expressing the carbapenem-hydrolysing oxacillinase OXA-58; Heritier C et al.; OBJECTIVE: The aim of this study was to analyse the spread of the bla(OXA-58) gene as a source of carbapenem resistance in Acinetobacter baumannii in the burns unit of a university hospital in Toulouse in 2003-2004 . METHODS: Six carbapenem-resistant A . baumannii isolates from six patients, and a carbapenem-resistant environmental A . baumannii isolate were collected in the burns unit of the Rangueil hospital (Toulouse) . Susceptibility tests were carried out by disc diffusion and agar dilution methods . The detection of the bla(OXA-58) gene was conducted by PCR followed by sequence analysis . Plasmids were extracted and hybridized with a probe specific for bla(OXA-58) . DNA fingerprints were obtained by pulsed-field gel electrophoresis of ApaI- or SmaI-digested chromosomal DNA of the tested strains . RESULTS: The multidrug-resistant clinical isolates had a similar 30 kb plasmid that encoded the carbapenem-hydrolysing beta-lactamase OXA-58 . These isolates were clonally related . The unrelated environmental carbapenem-resistant A . baumannii isolate had a similar bla(OXA-58)-carrying plasmid, suggesting spread of this gene . CONCLUSIONS: A novel oxacillinase was the source of carbapenem resistance in the A . baumannii isolates . Its gene was plasmid-, but not integron-borne. J Med Microbiol, 2004 Dec, 53(Pt 12), 1233 - 40 Diversity of aminoglycoside-resistance genes and their association with class 1 integrons among strains of pan-European Acinetobacter baumannii clones; Nemec A et al.; The purpose of the present study was to investigate the diversity of the genes encoding aminoglycoside-modifying enzymes and their association with class 1 integrons in three pan-European clones of Acinetobacter baumannii . The study collection included 106 multidrug-resistant strains previously allocated to clone I (n = 56), clone II (n = 36) and clone III (n = 6) and a heterogeneous group of other strains (n = 8), using AFLP fingerprinting and ribotyping . The strains were from hospitals of the Czech Republic (n = 70; collected 1991-2001) and 12 other European countries (n = 36; 1982-1998) . Using PCR, at least one of the following aminoglycoside-resistance genes was detected in 101 (95 %) strains: aphA1 (n = 76), aacC1 (n = 68), aadA1 (n = 68), aphA6 (n = 55), aadB (n = 31), aacC2 (n = 7) and aacA4 (n = 3) . A combination of two to five different resistance genes was observed in 89 strains (84 %), with a total of 12 different combinations . PCR mapping revealed that aacC1, aadA1 and aacA4 were each associated with a class 1 integron, as was the case with aadB for six strains of clone III . Six different class 1 integron variable regions were detected in 78 strains (74 %), with two predominant regions (2.5 and 3.0 kb) in two sets of 34 strains each . The 3.0 kb region contained five gene cassettes (aacC1, orfX, orfX, orfX', aadA1) and differed from the 2.5 bp region only by one additional orfX cassette . These two integron regions were confined to clones I and II and were found in strains isolated in seven countries between 1982 and 2001 . The clone III strains were homogeneous both in resistance genes and in integron variable regions, whereas clones I and II showed a remarkable intraclonal diversity of these properties, with no clear-cut difference between the two clones . Yet, within the Czech clone I and II strains, the diversity of resistance genes and integron structures was limited as compared to those from other countries . The occurrence of identical resistance genes, gene combinations and class 1 integrons associated with these genes in clonally distinct strains indicates that horizontal gene transfer plays a major role in the dissemination of aminoglycoside resistance in A . baumannii. Appl Environ Microbiol, 2004 Dec, 70(12), 7119 - 25 Synthesis of novel lipids in Saccharomyces cerevisiae by heterologous expression of an unspecific bacterial acyltransferase; Kalscheuer R et al.; The bifunctional wax ester synthase/acyl-coenzyme A:diacylglycerol acyltransferase (WS/DGAT) is the key enzyme in storage lipid accumulation in the gram-negative bacterium Acinetobacter calcoaceticus ADP1, mediating wax ester, and to a lesser extent, triacylglycerol (TAG) biosynthesis . Saccharomyces cerevisiae accumulates TAGs and steryl esters as storage lipids . Four genes encoding a DGAT (Dga1p), a phospholipid:diacylglycerol acyltransferase (Lro1p) and two acyl-coenzyme A:sterol acyltransferases (ASATs) (Are1p and Are2p) are involved in the final esterification steps in TAG and steryl ester biosynthesis in this yeast . In the quadruple mutant strain S . cerevisiae H1246, the disruption of DGA1, LRO1, ARE1, and ARE2 leads to an inability to synthesize storage lipids . Heterologous expression of WS/DGAT from A . calcoaceticus ADP1 in S . cerevisiae H1246 restored TAG but not steryl ester biosynthesis, although high levels of ASAT activity could be demonstrated for WS/DGAT expressed in Escherichia coli XL1-Blue in radiometric in vitro assays with cholesterol and ergosterol as substrates . In addition to TAG synthesis, heterologous expression of WS/DGAT in S . cerevisiae H1246 resulted also in the accumulation of fatty acid ethyl esters as well as fatty acid isoamyl esters . In vitro studies confirmed that WS/DGAT is capable of utilizing a broad range of alcohols as substrates comprising long-chain fatty alcohols like hexadecanol as well as short-chain alcohols like ethanol or isoamyl alcohol . This study demonstrated the highly unspecific acyltransferase activity of WS/DGAT from A . calcoaceticus ADP1, indicating the broad biocatalytic potential of this enzyme for biotechnological production of a large variety of lipids in vivo in prokaryotic as well as eukaryotic expression hosts. J Antimicrob Chemother, 2005 Jan, 55(1), 71 - 7 Epub 2004 Dec 01. The OPTAMA programme: utilizing MYSTIC (2002) to predict critical pharmacodynamic target attainment against nosocomial pathogens in Europe; Masterton RG et al.; OBJECTIVES: The Optimising Pharmacodynamic Target Attainment using the MYSTIC (Meropenem Yearly Susceptibility Test Information Collection) Antibiogram (OPTAMA) programme identifies antibiotic regimens with the highest probability of attaining critical pharmacodynamic targets, accounting for the inherent variability in pharmacokinetics, dosages and MIC distributions . METHODS: European MIC data were obtained from the MYSTIC programme . Pharmacodynamic target attainment was calculated by Monte Carlo simulation for meropenem, imipenem, ceftazidime, cefepime, piperacillin/tazobactam and ciprofloxacin against Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa . RESULTS: Significant differences in probability of target attainment were found, with Northern Europe demonstrating the highest probabilities of target attainment and Eastern Europe the lowest . The carbapenems had the highest target attainments and susceptibility levels across all regions and pathogens . The cephalosporins demonstrated high target attainments and susceptibility results against E . coli and K . pneumoniae in Northern and Southern Europe . Piperacillin/tazobactam and ciprofloxacin had the lowest levels for both parameters in all regions . Desirable target attainment was not achieved (except for carbapenems in Northern Europe) for A . baumannii and P . aeruginosa; thus, combination therapy may be appropriate empirical therapy for these pathogens in Southern and Eastern Europe . The closest correlations between target attainment and susceptibility were for meropenem 1 g every 8 h, imipenem 0.5 g every 6 h and ceftazidime 1 g every 8 h . CONCLUSIONS: The study highlighted significant overestimations between the probability of target attainment and the reported percentage susceptibility, particularly for piperacillin/tazobactam and ciprofloxacin . The approach of the OPTAMA programme provides a novel tool which complements susceptibility data to help in the selection of appropriate empirical antibiotic therapy. Vox Sang, 2004 Oct, 87(3), 143 - 9 Inactivation of Gram-negative and Gram-positive bacteria in red cell concentrates using INACTINE PEN110 chemistry; Zavizion B et al.; BACKGROUND AND OBJECTIVES: The risk of transfusion-transmitted bacterial infections as a result of the presence of bacteria in blood is one of the major concerns in transfusion medicine . The purpose of this study was to investigate whether bacteria inoculated into red blood cell concentrates can be inactivated by the INACTINE PEN110 pathogen-reduction process . Four bacterial species were chosen for the study: anaerobic Gram-positive Clostridium perfringens and Propionibacterium acnes, known to be transfusion-transmitted; and two Gram-negative species, Acinetobacter johnsonii and Acinetobacter lwoffii, recently reported to be a common cause of transfusion-associated infections in Europe . MATERIALS AND METHODS: Identical units of leucoreduced red cell concentrates were inoculated with A . johnsonii, A . lwoffii, C . perfringens, or P . acnes . The 4 degrees C control units were put on storage immediately after receiving the spike . The test units were subjected to PEN110 treatment and then stored . The bacterial titre in all units was monitored during a 6-week storage period . RESULTS: The PEN110 inactivation of all tested bacterial strains was time- and titre-dependent . For A . johnsonii and A . lwoffii, no viable bacteria were detected in the units spiked with up to 10(4) colony-forming units (CFU)/ml and treated with PEN110 . For red cell units spiked with 10(4)-10(5) CFU/ml of C . perfringens and P . acnes, no viable bacteria were detected in the units treated with PEN110 . In control units, there was a gradual decrease in A . johnsonii, A . lwoffii and C . perfringens titres during cold storage, while P . acnes titres remained stable . CONCLUSIONS: The PEN110 pathogen-reduction process was demonstrated to inactivate high titres of A . johnsonii, A . lwoffii, C . perfringens and P . acnes in red cell concentrates. S Afr Med J, 2004 Oct, 94(10 Pt 2), 857 - 61 Appropriate use of the carbapenems; Brink AJ et al.; The carbapenems are a group of broad-spectrum beta-lactam antibiotic agents of which there are three parenteral preparations currently available in South Africa, namely imimpenem/cilastatin, meropenem and ertapenem . Owing to the fact that imipenem/cilastatin and meropenem have a broad spectrum of activity that includes Pseudomonas and Acinetobacter species, they are ideal antibiotics for treatment of severe nosocomial infections . In contrast, ertapenem has limited in vitro activity against the latter non-fermentative gram-negative bacteria and is therefore more suitable for the treatment of certain severe community-acquired infections . This statement arises out of concerns about the general abuse of antibiotics such as the carbapenems, with the primary intention of highlighting the appropriate use of these agents. Infect Control Hosp Epidemiol, 2004 Nov, 25(11), 1002 - 4 Persistent Acinetobacter baumannii? Look inside your medical equipment; Bernards AT et al.; Two outbreaks of multidrug-resistant Acinetobacter baumannii occurred in our hospital . The outbreak strains were eventually isolated from respiratory ventilators, an apparatus used to cool or warm patients, and four continuous veno-venous hemofiltration machines . Removing dust from the machines and replacing all dust filters brought the outbreaks to an end. J Chemother, 2004 Oct, 16(5), 494 - 6 Multiresistant Stenotrophomonas maltophilia tunneled CVC-related sepsis, treated with systemic and lock therapy; Gattuso G et al.; In the last decade, a remarkable increase in the incidence of nosocomial Gram-negative infections has been observed . These pathogens represent a substantial problem in clinical practice, due to the high resistance profile of most commonly used antibiotics . This phenomenon is surely a co-factor that exposes these susceptible patients to infections caused by selected pathogens like multiresistant Gram-negative rods . A typical example is represented by VAP (ventilator-associated pneumonia) sustained by Acinetobacter spp., Pseudomonas aeruginosa, Bulkolderia cepacia . The Authors describe a case of a central venous cather (CVC)-related Stenotrophomonas maltophilia sepsis in a patient affected by solid tumor, successfully treated with systemic antibiotic therapy associated with "lock therapy" . This combination was able to cure the infection, allowing the patient to continue chemotherapy and saving the in situ CVC . The surveillance of CVCs, good adherence to the protocols and guidelines and "good practice" are the cornerstones for the prevention of nosocomial infections. J Hosp Infect, 2004 Dec, 58(4), 254 - 61 Antibiotic susceptibility and REP-PCR fingerprints of Acinetobacter spp . isolated from a hospital ten years apart; Misbah S et al.; The antibiotic susceptibility profiles and the repetitive extragenic palindromic sequence-based polymerase chain reaction (REP-PCR)-determined genotypes of 109 Acinetobacter strains collected from the University Malaya Medical Center (UMMC), Kuala Lumpur, Malaysia, in 1987 (N=21) and 1996-1998 (N=88) were established . Twelve antibiotic susceptibility profiles of antibiotics used at the UMMC were obtained . In descending order of effectiveness, imipenem, amikacin and ciprofloxacin were the most effective against the Acinetobacter strains . Compared with 1987 isolates, the isolates obtained in 1996-1998 had decreased susceptibility to these antibiotics and were tolerant to the antibiotics up to an MIC90 of > or =256 mg/L . REP-PCR DNA fingerprints of all the isolates revealed the presence of four Acinetobacter spp . lineages; 92% of all the isolates belonged to two dominant lineages (genotypes 1 and 4) . Genotype 4 isolates predominant in 1987 showed increased resistance and antibiotic tolerance to imipenem, amikacin and ciprofloxacin compared with the 1996-1998 isolates . In contrast, genotype 1 isolates from 1996-1998 were mainly sensitive to these antibiotics . These findings demonstrate the presence of at least two independent Acinetobacter spp . lineages in the same hospital, and suggest the possibility that genotype 4 Acinetobacter spp . acquired the resistance phenotype in situ, whereas most of the genotype 1 isolates were probably introduced to the hospital in recent years. Antimicrob Agents Chemother, 2004 Dec, 48(12), 4919 - 21 The synthetic N-terminal peptide of human lactoferrin, hLF(1-11), is highly effective against experimental infection caused by multidrug-resistant Acinetobacter baumannii; Dijkshoorn L et al.; The lactoferrin-derived peptide hLF(1-11), but not its control peptide, was highly effective against five multidrug-resistant Acinetobacter baumannii strains in vitro (3 to 4 log reduction) and against four of these strains in an experimental infection in mice (2 to 3 log reduction) . Therefore, this peptide is a promising candidate as a novel agent against infections with multidrug-resistant A . baumannii. Transplant Proc, 2004 Oct, 36(8), 2377 - 9 Heart transplantation in the patient under ventricular assist complicated with device-related infection; Wang SS et al.; Ventricular assist devices (VAD) have benefited patients with end-stage heart failure as a bridge to heart transplantation (HTx) . We present our experience of HTx in the presence of device-related infection (DRI) including driveline exit site with pocket infections . From May 1996 to April 2003, mechanical circulatory support with the HeartMate VAD was performed in eight patients, and with the Thoratec VAD in seven patients . Although 151 patients underwent HTx during that period, only 8 of the 15 patients had suitable donors and underwent orthotopic HTx . Six of the eight patients developed DRI . Their ages ranged from 18 to 59 years (mean = 36 +/- 14 years) . The duration of VAD support ranged from 8 to 287 days (mean = 125 +/- 117 days) . The general condition and cardiac function improved gradually under VAD support . At the time of HTx, all six male patients were suffering from DRI . The causative microorganisms were Acinetobacter baumannii (n = 3) methicillin-resistant Staphylococcus aureus (n = 2), and Enterococcus faecium (n = 1) . All patients underwent successful HTx, and were discharged in good condition . It is concluded that under the coverage of appropriate antibiotics, HTx can be successfully performed for the patients for VAD support with DRI . It is important to prevent the spread of infection during HTx . Adequate debridement and drainage of the infected materials prevents postoperative wound infections. Environ Microbiol, 2004 Dec, 6(12), 1244 - 51 Isolation and characterization of soybean-associated bacteria and their potential for plant growth promotion; Kuklinsky-Sobral J et al.; Endophytic and epiphytic bacteria were isolated from two soybean cultivars (Foscarin and Cristalina) . Significant differences were observed in bacterial population densities in relation to season of isolation, soybean growth phase and the tissues from which the isolates were obtained . The isolates were identified by partial 16S rDNA sequence analysis, with most of the isolates belonging to the Pseudomonaceae, Burkholderiacea and Enterobacteriaceae groups . The potential of the isolates for plant growth promotion was evaluated by screening for indoleacetic acid (IAA) production and mineral phosphate solubilization; 34% of endophytic bacteria produced IAA and 49% were able to solubilize mineral phosphate whereas only 21% of epiphytic bacteria produced IAA although 52% were able to solubilize mineral phosphate . A high frequency of IAA producing isolates occurred in the early ripening Foscarin cultivar whereas a high percentage of phosphate solubilizing isolates were obtained from plants in the initial development stage (V6) . We also found that 60% of endophytic and 69% of epiphytic isolates that produced IAA and solubilized mineral phosphate were also able to fix nitrogen in vitro . The soybean-associated bacteria showing characteristics related to plant growth promotion were identified as belonging to the genera Pseudomonas, Ralstonia, Enterobacter, Pantoea and Acinetobacter. Klin Mikrobiol Infekc Lek, 2004 Oct, 10(5), 223 - 8 {Aminoglycoside resistance of Acinetobacter baumannii hospital strains in the Czech Republic.}; Nemec A et al.; OBJECTIVE: To investigate the nature of aminoglycoside resistance of clinically important multidrug-resistant (MDR) Acinetobacter baumannii strains from the Czech Republic . MATERIAL AND METHODS: Seventy MDR hospital strains from 1991-2001 classified into pan-European clones I (n = 41) and II (n = 21) and a group of other MDR strains (n = 8) were studied . Fifteen other strains wild-type susceptible to aminoglycosides were used as controls . Susceptibility to aminoglycosides was determined by the disk diffusion and agar dilution tests . The presence of six genes encoding different aminoglycoside-modifying enzymes (AME) and of the adeB gene encoding a component of the AdeABC efflux pump was detected using PCR . RESULTS: MDR strains showed the following pattern of resistance to aminoglycosides: kanamycin (93 %; inhibition zone diameters < or =13 mm), gentamicin (87 %; MICs > or =16 mg/l), amikacin (47 %; MICs > or =64 mg/l), tobramycin (31 %; MICs > or = 16 mg/l) and netilmicin (17 %; MICs ? 32 mg/l) . Forty-six (66 %) MDR strains had netilmicin MICs of 8-16 mg/l in contrast to 0.5-1.0 mg/l found for control susceptible strains . The presence of at least one of the following AME genes was detected in 66 (94 %) MDR strains: aphA1 (n = 56), aacC1 (n = 52), aphA6 (n = 39) and aadB (n = 20) . The presence of these genes and phenotypes of resistance to kanamycin, gentamicin, tobramycin and amikacin were in good agreement . The genes encoding netilmicin-modifying enzymes (aacC2, aacA4) were not detected in any strain . Fifty-six (80 %) MDR strains comprised two to four different AME genes in eight combinations . Clones I and II shared all of the genes studied (with the exception of aadB not detected in clone I) . The gene adeB was found in all MDR strains and eight of 15 control susceptible strains . CONCLUSION: Clinically relevant aminoglycoside resistance of Czech A . baumannii strains is significantly associated with the genes encoding enzymatic modification of kanamycin, gentamicin, amikacin and tobramycin . These genes can spread horizontally and emerge in different combinations leading to high-level resistance to multiple aminoglycosides . The AdeABC pump is likely to play a role in the intermediate susceptibility to netilmicin but further study is needed in this regard. Eur J Clin Microbiol Infect Dis, 2004 Dec, 23(12), 888 - 91 Nosocomial lower respiratory tract infections: prevalence and risk factors in 14 Greek hospitals; Kofteridis DP et al.; Nosocomial lower respiratory tract infections (NLRTIs) are associated with significant morbidity and mortality . The aim of this study was to investigate the epidemiological features of NLRTIs in Greece, where knowledge about these infections is limited . Two point-prevalence studies of hospital-acquired infections were carried out in 14 Greek hospitals located throughout the country, one in 1999 and one in 2000 . NLRTIs were diagnosed in accordance with the Centers for Disease Control and Prevention (CDC) definitions . Among the 7,120 hospitalized patients registered during the two studies, 610 (8.6%) cases of hospital-acquired infections were identified, of which 200 (32.8%) were NLRTIs . Sixty-nine (34.5%) patients had pneumonia, and the remaining 131 (65.5%) patients had bronchitis . The greatest prevalence of NLRTI was found in the adult ICUs (30.4%) . Male gender, age >65 years, mechanical ventilation, tracheostomy, an intravenous central line, and an indwelling urethral catheter were the main risk factors . There was no significant difference in the incidence of NLRTI among hospital-acquired infections between the 1999 study and the 2000 study . The causative microorganism was identified in 78 of 200 (39%) cases, and 103 strains were isolated . The majority of strains (67%) were gram-negative bacteria . The most frequently isolated microorganisms were Pseudomonas aeruginosa (22.3%), Acinetobacter spp . (19.4%), Klebsiella pneumoniae (12.6%), and Staphylococcus aureus (10.7%) . There was no difference between the two prevalence studies in the frequency of isolation of the microorganisms . NLRTI was the leading cause of morbidity and mortality among hospitalized patients with hospital-acquired infections in Greek hospitals . Gram-negative microorganisms were the most frequently isolated pathogens. Biosens Bioelectron, 2004 Dec 15, 20(6), 1217 - 22 Wiring of PQQ-dehydrogenases; Laurinavicius V et al.; The performance of pyrroloquinoline quinone (PQQ) dependent alcohol dehydrogenase (ADH) and two types of PQQ-glucose dehydrogenases in solution and when immobilized on the carbon paste electrodes modified with ferrocene derivatives is investigated . The immobilization of ADH consisting of PQQ and four hemes improves its stability up to 10 times . Both PQQ and heme moieties are involved in the electron transport from substrate to electrode . The ferrocene derivatives improve the electron transport 10-fold . Membrane-bound alcohol dehydrogenase from Gluconobacter sp . 33, intracellular soluble glucose dehydrogenase from Acinetobacter calcoaceticus L.M.D . 79.41 (s-GDH), and the membrane-bound enzyme (m-GDH) from Erwinia sp . 34-1 were purified and investigated . Soluble and membrane-bound PQQ-glucose dehydrogenases display different behavior during the immobilization on the modified carbon electrodes . The immobilization of s-GDH leads to a decrease in both stability and substrate specificity of the enzyme . This suggests that PQQ dissociates from the enzyme active center and operates as a free-diffusing mediator . The rate-limiting step of the process is likely the loading of PQQ onto the apo-enzyme . The immobilization of m-GDH leads to its substantial stabilization and improves the substrate specificity . The nature of m-GDH binding to the electrode surface is presumably similar to the binding to the cell membrane through its anchor-subunit . The enzyme operates as an enzyme and mediator complex. Ann Ig, 2004 Jan-Apr, 16(1-2), 95 - 102 Clonal spread of Acinetobacter baumannii in a general intensive care unit; Marroni M et al.; The epidemiological characterization of multiply resistant Acinetobacter baumannii isolates from a six-bed Intensive Care Unit (ICU) is described . Investigations for A . baumannii were performed in three subsequent surveillance studies . In the first study, surveillance cultures were taken from patients, health care personnel and the environment; in the second study surveillance cultures were taken at 0, 4, and 7 days from all patients admitted consecutively to the ward; and in the third study surveillance cultures were taken from patients, health care personnel and the environment . During the first study all four hospitalized patients were found to harbour A . baumannii . Hand cultures did not grow any A . baumannii when staff entered the ward from home, but 7 positive health care workers were identified out of 25 samples taken during work, and two cultures of environmental specimens grew A . baumannii . During the second study, 4 of 86 (4.6%) patients resulted colonized with A . baumannii . In the third epidemiological study, no A . baumannii was cultured from either patients, health care personnel or the environment . All isolates recovered from various patients or sources produced conserved macrorestriction Pulsed-Field Gel Electrophoresis (PFGE) patterns and showed the same antibiotic resistance; therefore, they can be considered indistinguishable . The same antibiotic resistance and macrorestriction patterns were observed in previously isolated A . baumannii strains in the ward during May 1997, suggesting the persistence of a single A . baumannii in the ICU . The present study confirms that molecular typing is an essential tool in the epidemiology and control of nosocomial infections, showing here the persistence of a single A . baumannii clone in the ICU . The origin of this strain remains unknown but, when basic infection control measures were reinforced, emphasizing the importance of hand antisepsis and judicious use of gloves, control of A . baumannii spread in the ward was achieved. Vojnosanit Pregl, 2004 Jul-Aug, 61(4), 391 - 7 {The frequency of resistance to antibiotics of most frequently isolated bacteria from blood cultures during the period 1997-2002}; Mirovic V et al.; The aim of this study was to determine the frequency of resistance to antibiotics of the most frequently isolated bacteria from blood cultures of hospitalized patients during the period 1997-2002 . The resistance to antibiotics was determined by disk diffusion method according to National Committee for Clinical Laboratory Standards procedures . The majority of staphylococci isolates were resistant to methicillin, and the proportion of methicillin-resistant Staphylococcus aureus was stable (76.8-81.6%), during the follow-up period . None of the staphylococci isolates were resistant to vancomycin, but there was a very high incidence of high-level resistance of enterococci to aminoglycosides (47.2-72.2%) . In 1998, only one strain among enterococci was resistant to vancomycin (Enterococcus faecium, VanA phenotype) . Enterococcus spp isolates expressed variable frequency of resistance to ampicillin (15-40.1%) during the follow-up period . Among Enterobacteriaceae there were no isolates resistant to imipenem, but dramatic increase of the resistance to ceftriaxone was found from 35.9% in 1997 to 95.9% in 2002 (p<0.001) . Extended spectrum beta-lactamases production was found in all the species of enterobacteria isolates . Resistance to imipenem was observed in Acinetobacter spp isolates in 2002 for the first time . Pseudomonas spp isolates expressed high and very variable resistance to all antibiotics tested during the follow-up period. MMWR Morb Mortal Wkly Rep, 2004 Nov 19, 53(45), 1063 - 6 Acinetobacter baumannii infections among patients at military medical facilities treating injured U.S . service members, 2002-2004; Centers for Disease Control and Prevention (CDC); Acinetobacter baumannii is a well known but relatively uncommon cause of health-care--associated infections . Because the organism has developed substantial antimicrobial resistance, treatment of infections attributed to A . baumannii has become increasingly difficult This report describes an increasing number of A . baumannii bloodstream infections in patients at military medical facilities in which service members injured in the Iraq/Kuwait region during Operation Iraqi Freedom (OIF) and in Afghanistan during Operation Enduring Freedom (OEF) were treated . The number of these infections and their resistance to multiple antimicrobial agents underscore 1) the importance of infection control during treatment in combat and health-care settings and 2) the need to develop new antimicrobial drugs to treat these infections. J Antimicrob Chemother, 2004 Dec, 54(6), 1085 - 91 Epub 2004 Nov 16. Antibiotic combinations for serious infections caused by carbapenem-resistant Acinetobacter baumannii in a mouse pneumonia model; Montero A et al.; OBJECTIVES: Successful therapy of carbapenem-resistant Acinetobacter baumannii strains has been reported with colistin, but recently we argued against its use as monotherapy because of the poor results obtained in a mouse pneumonia model . Our aim was to identify antibiotic combinations that were valid therapeutic alternatives in the same model . METHODS: We used two carbapenem-resistant A . baumannii strains (D and E; MICs of imipenem, 8 and 512 mg/L, respectively) . MICs of tobramycin, rifampicin and colistin for both strains were 8, 8 and 0.5 mg/L, respectively . RESULTS: In infections caused by strain D, lung bacterial counts (log(10) cfu/g, mean +/- s.d.) were: controls (10.86+/-0.25), imipenem (5.99+/-0.59, P < 0.05 versus controls), and colistin (10.43 +/- 1.09); imipenem + tobramycin was the most active combination (5.46+/-0.62, P < 0.05 versus controls) . In infections caused by strain E, results were: controls (10.82+/-0.33), rifampicin (5.62+/-0.26, P < 0.05 versus controls), colistin (8.38+/-1.22, P < 0.05 versus controls), and imipenem (11.01+/-0.2); rifampicin + imipenem (3.79+/-0.99) and rifampicin + tobramycin (3.96+/-0.30) were the most active combinations (P < 0.05); results with rifampicin + colistin (5.59+/-1.17) were similar to those with rifampicin alone . CONCLUSIONS: Our data indicate that imipenem can still be the best alternative for carbapenem-resistant A . baumannii infections with moderate levels of imipenem resistance, preferably combined with aminoglycosides . For strains highly resistant to imipenem, a combination of rifampicin with imipenem, tobramycin or colistin may be useful, if resistance to rifampicin is only moderate. Med Dosw Mikrobiol, 2004, 56(2), 179 - 85 {Adhesion of Acinetobacter spp . to para-xylene}; Krasnicki K et al.; Hydrophobicity of 200 of Acinetobacter spp . strains was defined by the BATH (Bacterial Adhesion to Hydrocarbon Test) method of Rosenberg et al . 48.0% strains of Acinetobacter spp . adhered to para-xylene . 40.0% of strains showed waek hydrophobicity, 7.5% moderate and one strain strongly adhered to para-xylene . A . baumannii strains adhered in 52.2%, A . junii in 42.1% and 20.0% in A . haemolyticus . Significantly we find weak adhesion to para-xylene than mild and strong . Mostly adhesion was discovered in A . baumannii and A . junii than in A . haemolyticus . Results show that Acinetobacter spp . has adhesive properties to para-xylene. J Clin Microbiol, 2004 Nov, 42(11), 5256 - 63 Metallo-beta-lactamase-producing gram-negative bacilli: laboratory-based surveillance in cooperation with 13 clinical laboratories in the Kinki region of Japan; Nishio H et al.; A total of 19,753 strains of gram-negative rods collected during two 6-month periods (October 2000 to March 2001 and November 2001 to April 2002) from 13 clinical laboratories in the Kinki region of Japan were investigated for the production of metallo-beta-lactamases (MBLs) . MBLs were detected in 96 (0.5%) of the 19,753 isolates by the broth microdilution method, the 2-mercaptopropionic acid inhibition test, and PCR and DNA sequencing analyses . MBL-positive isolates were detected in 9 of 13 laboratories, with the rate of detection ranging between 0 and 2.6% for each laboratory . Forty-four of 1,429 (3.1%) Serratia marcescens, 22 of 6,198 (0.4%) Pseudomonas aeruginosa, 21 of 1,108 (1.9%) Acinetobacter spp., 4 of 544 (0.7%) Citrobacter freundii, 3 of 127 (2.4%) Providencia rettgeri, 1 of 434 (0.2%) Morganella morganii, and 1 of 1,483 (0.1%) Enterobacter cloacae isolates were positive for MBLs . Of these 96 MBL-positive strains, 87 (90.6%), 7 (7.3%), and 2 (2.1%) isolates carried the genes for IMP-1-group MBLs, IMP-2-group MBLs, and VIM-2-group MBLs, respectively . The class 1 integrase gene, intI1, was detected in all MBL-positive strains, and the aac (6')-Ib gene was detected in 37 (38.5%) isolates . Strains with identical PCR fingerprint profiles in a random amplified polymorphic DNA pattern analysis were isolated successively from five separate hospitals, suggesting the nosocomial spread of the organism in each hospital . In conclusion, many species of MBL-positive gram-negative rods are distributed widely in different hospitals in the Kinki region of Japan . The present findings should be considered during the development of policies and strategies to prevent the emergence and further spread of MBL-producing bacteria. J Clin Microbiol, 2004 Nov, 42(11), 5102 - 8 Antimicrobial susceptibility testing of Acinetobacter spp . by NCCLS broth microdilution and disk diffusion methods; Swenson JM et al.; Although both broth microdilution (BMD) and disk diffusion (DD) are listed by NCCLS as acceptable methods for testing Acinetobacter spp . for antimicrobial susceptibility, few studies have compared the results generated by the two methods . We tested 196 isolates of Acinetobacter spp . from nine U.S . hospitals and from the Centers for Disease Control culture collection by using BMD and DD and clinically appropriate antimicrobial agents . Categorical results for amikacin, ciprofloxacin, gatifloxacin, gentamicin, imipenem, levofloxacin, meropenem, tobramycin, and trimethoprim-sulfamethoxazole were comparable for the two methods: there was only one very major (VM) error, with tobramycin, and only one major (M) error, with meropenem, when DD results were compared with BMD results . However, VM errors were frequent with the beta-lactams and beta-lactam-beta-lactam inhibitor combinations, while M errors were often observed with tetracyclines . For BMD, tests frequently exhibited subtle growth patterns that were difficult to interpret, especially for beta-lactams . If subtle growth (i.e., granular, small button, or "starry" growth) was considered positive, error rates between BMD and DD were unacceptably high for ampicillin-sulbactam (VM error, 9.8%; minor {m} error, 16.1%), piperacillin (VM error, 5.7%; m error, 13.5%), piperacillin-tazobactam (VM error, 9.3%; m error, 12.9%), ceftazidime (VM error, 6.2%; m error, 11.4%), cefepime (VM error, 6.2%; m error, 13.0%), cefotaxime (m error, 21.2%), ceftriaxone (m error, 23.3%), tetracycline (M error, 11.4%; m error, 32.1%), and doxycycline (M error, 2.6%) . When subtle growth patterns were ignored, the agreement still did not achieve acceptable levels . To determine if the problems with BMD testing occurred in other laboratories, we sent frozen BMD panels containing beta-lactam drugs and nine isolates to six labs with experience in performing BMD and DD . Among these laboratories, cefepime MICs ranged from < or =8 to > or =32 microg/ml for four of the nine strains, confirming the problem in interpreting BMD results . Discrepancies between the categorical interpretations of BMD and DD tests were noted primarily with cefepime and piperacillin, for which the BMD results were typically more resistant . Clinical laboratories should be aware of these discrepancies . At present, there are no data to indicate which method provides more clinically relevant information. Microbiology, 2004 Nov, 150(Pt 11), 3657 - 67 The siderophore-mediated iron acquisition systems of Acinetobacter baumannii ATCC 19606 and Vibrio anguillarum 775 are structurally and functionally related; Dorsey CW et al.; The Acinetobacter baumannii type strain, ATCC 19606, secretes acinetobactin, a catechol siderophore highly related to the iron chelator anguibactin produced by the fish pathogen Vibrio anguillarum (Listonella anguillarum) . This paper reports the initial characterization of the genes and gene products involved in the acinetobactin-mediated iron-acquisition process . Insertional mutagenesis resulted in the isolation of several derivatives whose ability to grow in medium containing the iron chelator 2,2'-dipyridyl was affected . One of the insertions disrupted a gene encoding a predicted outer-membrane protein, named BauA, highly similar to FatA, the receptor for ferric anguibactin . Immunological relatedness of BauA with FatA was confirmed by Western blot analysis . Another transposon insertion was mapped to a gene encoding a protein highly similar to FatD, the permease component of the anguibactin transport system . Further DNA sequencing and nucleotide sequence analysis revealed that these A . baumannii 19606 genes are part of a polycistronic locus that contains the bauDCEBA ORFs . While the translation products of bauD, -C, -B and -A are highly related to the V . anguillarum FatDCBA iron-transport proteins, the product of bauE is related to the ATPase component of Gram-positive ATP-binding cassette (ABC) transport systems . This entire locus is flanked by genes encoding predicted proteins related to AngU and AngN, V . anguillarum proteins required for the biosynthesis of anguibactin . These protein similarities, as well as the structural similarity of anguibactin and acinetobactin, suggested that these two siderophores could be utilized by both bacterial strains, a possibility that was confirmed by siderophore utilization bioassays . Taken together, these results demonstrate that these pathogens, which cause serious infections in unrelated hosts, express very similar siderophore-mediated iron-acquisition systems. Clin Microbiol Infect, 2004 Nov, 10 Suppl 4, 1 - 9 The need for new antibiotics; Livermore DM; Politicians and public health officials have joined specialist professionals in recognising antibiotic resistance as a threat to modern medicine . Their response has centred on minimising unnecessary antibiotic prescribing, aiming to reduce selection pressure for resistance . Despite a few hopeful trends (e.g., declining penicillin resistance among pneumococci in the UK), established resistance is proving hard to displace; moreover, new resistances continue to emerge and to proliferate at new sites . There consequently remains a strong need for new antibiotics, particularly those directed against multiresistant Gram-negative bacteria in hospitals . Already some nonfermenters of the genera Acinetobacter and Pseudomonas are resistant to all good antibiotics and many Enterobacteriaceae are resistant to all except carbapenems . There is also a growing need for new agents against community-acquired pathogens, including the agents of tuberculosis, gonorrhoea and urinary tract infections . Unless antibacterial development is re-energised, there is a serious risk that a growing proportion of infections, especially in hospitals, will become effectively untreatable. J Med Assoc Thai, 2004 Jul, 87(7), 819 - 24 Device-associated infections and patterns of antimicrobial resistance in a medical-surgical intensive care unit in a university hospital in Thailand; Thongpiyapoom S et al.; BACKGROUND: Surveillance of nosocomial infection in the intensive care unit (ICU) received a high level of attention and outcome indicators are now used in benchmarking the quality of patient care . Since 1999 the surveillance has targeted three site-specific, device-associated infections, including ventilator-associated pneumonia (VAP), central-line-related bloodstream infection (CR-BSI), and catheter-related urinary tract infection (CR-UTI) . The authors conducted a two-year prospective study on the incidences of these infections acquired in an ICU and report herein, together with the antibiotic susceptibility patterns of the microorganisms isolated in an ICU . PATIENTS AND METHOD: Continuous prospective data collection was conducted on patients admitted to an adult medical-surgical ICU of a university hospital in Thailand from June 2000 to May 2002 . RESULTS: A total 1422 patients with a total of 9370 patient-days were enrolled in the study . The incidence of VAP, CR-BSI, and CR-UTI were 10.8/1000 ventilator-days (95% C.I: 8.5-13.6), 2.6/1000 central-line-days (95%C.I . 1.5-4.4), and 13.8/1000 urinary-catheter-days (95%C.I: 10.7-17.5) respectively . The most common causative pathogens were Escherichia coli, Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae . The proportion of methicillin-resistant Staphylococcus aureus, imipenem-resistant P . aeruginosa, ceftazidime-resistant A . baumannii, third-generation-cephalosporin-resistant K . pneumoniae, and quinolone-resistant E . coli were 68.8%, 30.9%, 68.5%, 44.6%, 38.3% respectively . CONCLUSION: The incidences of VAP and CR-BSI were comparable to the National Nosocomial Infection Surveillance (NNIS) report . But the incidence of CR-UTI was over the 90th percentile . The antibiotic resistance had become a serious problem. Infect Control Hosp Epidemiol, 2004 Oct, 25(10), 868 - 72 Emergence of resistance in Pseudomonas aeruginosa and Acinetobacter species after the use of antimicrobials for burned patients; Ferreira AC et al.; OBJECTIVES: To evaluate the emergence of resistance of Pseudomonas aeruginosa and Acinetobacter species to imipenem, ciprofloxacin, or both after the use of these drugs and to compare resistant with susceptible isolates by molecular typing . DESIGN: Cohort study . SETTING: Burn intensive care unit (ICU) with 4 beds in a tertiary-care university hospital . METHODS: During 16 months, surveillance cultures were performed for all patients admitted to the ICU . Demographic information was obtained for each patient . Molecular typing was done by pulsed-field gel electrophoresis using restriction enzymes for 71 isolates of P . aeruginosa and Acinetobacter species . RESULTS: Thirty-four patients were admitted and 22 were colonized by susceptible P . aeruginosa or Acinetobacter species before they used the antimicrobials . Nine (41%) of these patients had a resistant isolate after antimicrobial use: 5 had used imipenem alone, 1 had used ciprofloxacin, and 3 had used both drugs . The interval between isolation of the susceptible and resistant isolates ranged from 4 to 25 days, but was 10 or more days for 6 patients . Molecular typing revealed that susceptible and resistant isolates from each patient were different and that although there were no predominant clones among susceptible isolates, there was a predominant clone among resistant isolates of P . aeruginosa and of Acinetobacter . CONCLUSIONS: Resistance was not due to the acquisition of resistance mechanisms by a previously susceptible strain, but rather to cross-transmission . Although various measures involving antimicrobial use have received great attention, it would seem that practices to prevent cross-transmission are more important in controlling resistance. Infect Control Hosp Epidemiol, 2004 Oct, 25(10), 847 - 51 Evaluation of three molecular typing techniques for nonfermentative Gram-negative bacilli; Silbert S et al.; OBJECTIVE: To evaluate three different DNA techniques for typing nonfermentative gram-negative bacilli isolated from Latin American hospitals . DESIGN: One hundred twenty-six nonfermentative gram-negative bacilli were typed . PARTICIPANTS: Pseudomonas aeruginosa (n = 64) and Acinetobacter baumannii (n = 42) samples were obtained from blood cultures of patients admitted to 10 medical centers in Latin America during 1998 and Stenotrophomonas maltophilia (n = 20) samples were obtained from patients admitted to the Hospital Sao Paulo between 1999 and 2001 . METHODS: All samples were typed using automated ribotyping, PFGE, and ERIC-PCR . The discriminatory power for each technique was calculated using Hunter's generalized formula . RESULTS: All strains could be typed by automated ribotyping and ERIC-PCR, but two strains (1.6%) were not typeable by PFGE . All three techniques showed 100% reproducibility . The time to obtain the results was shorter for automated ribotyping and ERIC-PCR compared with PFGE . Likewise, the costs for ERIC-PCR and PFGE were lower than those for automated ribotyping . The interpretation of results was more complicated and more difficult with ERIC-PCR than with both PFGE and automated ribotyping . All techniques presented excellent discriminatory power for P . aeruginosa (0.98) . PFGE presented the highest discriminatory power (0.94) for A . baumannii, and both PFGE and ERIC-PCR showed higher discriminatory power (0.90 for both) than automated ribotyping (0.82) for S . maltophilia . CONCLUSIONS: PFGE showed the highest discriminatory power for typing these nonfermentative gram-negative bacilli . However, automated ribotyping and ERIC-PCR can provide results in a shorter time period with similar discriminatory power. Infect Control Hosp Epidemiol, 2004 Oct, 25(10), 819 - 24 Clinical features and epidemiology of Acinetobacter baumannii colonization and infection in Spanish hospitals; Rodriguez-Bano J et al.; OBJECTIVE: To investigate the clinical features and the epidemiology of Acinetobacter baumannii in Spanish hospitals . DESIGN: Prospective multicenter cohort study . SETTING: Twenty-seven general hospitals and one paraplegic center in Spain . METHODS: All cases of A . baumannii colonization or infection detected by clinical samples during November 2000 were included . Isolates were identified using phenotypic and genotypic methods . The molecular relatedness of the isolates was assessed by pulsed-field gel electrophoresis . RESULTS: Twenty-five (89%) of the hospitals had 221 cases (pooled rate in general hospitals, 0.39 case per 1,000 patient-days; range, 0 to 1.17) . The rate was highest in intensive care units (ICUs) . Only 3 cases were pediatric . The mean age of the patients in the general hospitals was 63 years; 69% had a chronic underlying disease and 80% had previously received antimicrobial treatment . Fifty-three percent of the patients had an infection (respiratory tract, 51%; surgical site, 16%; and urinary tract, 11%) . Crude mortality was higher in infected than in colonized patients (27% vs 10%; relative risk, 1.56; 95% confidence interval, 1.2 to 2.0; P = .003) . Molecular analysis disclosed 79 different clones . In most hospitals, a predominant epidemic clone coexisted with other sporadic clones . Imipenem resistance was present in 39% of the hospitals . CONCLUSIONS: A . baumannii was present in most participating Spanish hospitals (particularly in ICUs) with different rates among them . The organisms mainly affected predisposed patients; half of them were only colonized . Epidemic and sporadic clones coexisted in many centers. Nucleic Acids Res, 2004 Oct 28, 32(19), 5780 - 90 Print 2004. Acinetobacter sp . ADP1: an ideal model organism for genetic analysis and genome engineering; Metzgar D et al.; Acinetobacter sp . strain ADP1 is a naturally transformable gram-negative bacterium with simple culture requirements, a prototrophic metabolism and a compact genome of 3.7 Mb which has recently been sequenced . Wild-type ADP1 can be genetically manipulated by the direct addition of linear DNA constructs to log-phase cultures . This makes it an ideal organism for the automation of complex strain construction . Here, we demonstrate the flexibility and versatility of ADP1 as a genetic model through the construction of a broad variety of mutants . These include marked and unmarked insertions and deletions, complementary replacements, chromosomal expression tags and complex combinations thereof . In the process of these constructions, we demonstrate that ADP1 can effectively express a wide variety of foreign genes including antibiotic resistance cassettes, essential metabolic genes, negatively selectable catabolic genes and even intact operons from highly divergent bacteria . All of the described mutations were achieved by the same process of splicing PCR, direct transformation of growing cultures and plating on selective media . The simplicity of these tools make genetic analysis and engineering with Acinetobacter ADP1 accessible to laboratories with minimal microbial genetics expertise and very little equipment . They are also compatible with complete automation of genetic analysis and engineering protocols. Nucleic Acids Res, 2004 Oct 28, 32(19), 5766 - 79 Print 2004. Unique features revealed by the genome sequence of Acinetobacter sp . ADP1, a versatile and naturally transformation competent bacterium; Barbe V et al.; Acinetobacter sp . strain ADP1 is a nutritionally versatile soil bacterium closely related to representatives of the well-characterized Pseudomonas aeruginosa and Pseudomonas putida . Unlike these bacteria, the Acinetobacter ADP1 is highly competent for natural transformation which affords extraordinary convenience for genetic manipulation . The circular chromosome of the Acinetobacter ADP1, presented here, encodes 3325 predicted coding sequences, of which 60% have been classified based on sequence similarity to other documented proteins . The close evolutionary proximity of Acinetobacter and Pseudomonas species, as judged by the sequences of their 16S RNA genes and by the highest level of bidirectional best hits, contrasts with the extensive divergence in the GC content of their DNA (40 versus 62%) . The chromosomes also differ significantly in size, with the Acinetobacter ADP1 chromosome <60% of the length of the Pseudomonas counterparts . Genome analysis of the Acinetobacter ADP1 revealed genes for metabolic pathways involved in utilization of a large variety of compounds . Almost all of these genes, with orthologs that are scattered in other species, are located in five major 'islands of catabolic diversity', now an apparent 'archipelago of catabolic diversity', within one-quarter of the overall genome . Acinetobacter ADP1 displays many features of other aerobic soil bacteria with metabolism oriented toward the degradation of organic compounds found in their natural habitat . A distinguishing feature of this genome is the absence of a gene corresponding to pyruvate kinase, the enzyme that generally catalyzes the terminal step in conversion of carbohydrates to pyruvate for respiration by the citric acid cycle . This finding supports the view that the cycle itself is centrally geared to the catabolic capabilities of this exceptionally versatile organism. Antimicrob Agents Chemother, 2004 Nov, 48(11), 4479 - 81 Activity of tigecycline (GAR-936) against Acinetobacter baumannii strains, including those resistant to imipenem; Pachon-Ibanez ME et al.; We determined the in vitro activities of tigecycline and imipenem against 49 isolates of Acinetobacter baumannii, including those resistant to imipenem . The MIC at which 50% of the isolates were inhibited (MIC(50)) and the MIC(90) for tigecycline and imipenem were 2 and 2 mg/liter and 32 and 128 mg/liter, respectively, with 92 and 20%, respectively, of the strains being susceptible . Tigecycline did not show bactericidal activity in the time-kill studies (n = 9 strains) . Imipenem showed bactericidal activity against seven out of nine strains . These in vitro results show that tigecycline has good in vitro bacteriostatic activity against A . baumannii, including strains resistant to imipenem. J Hosp Infect, 2004 Nov, 58(3), 170 - 9 A prevalent, multiresistant clone of Acinetobacter baumannii in Southeast England; Turton JF et al.; A multiresistant clone of Acinetobacter baumannii was identified in 24 hospitals in the UK, predominantly in the London area, over a period of three years . Isolates were characterized by distinctive ApaI macrorestriction profiles, as resolved by pulsed-field gel electrophoresis (PFGE), which all clustered within 80% similarity using a 1% band position tolerance setting . The first isolates identified were received by the reference laboratories in April 2000, and by June 2003, a total of 375 isolates with similar PFGE profiles from 310 patients from 24 hospitals had been received . The isolates originated mainly from sputum and wound specimens, with the majority from patients in intensive care units . Amplified fragment length polymorphism analysis of a subset of isolates showed that they clustered closely, supporting the PFGE results . All the isolates tested were highly resistant to ampicillin, piperacillin, piperacillin/tazobactam, ceftazidime, cefotaxime, gentamicin and ciprofloxacin, and most isolates were carbapenem resistant . Amikacin sensitivity varied from susceptible {minimum inhibitory concentration (MIC) <or= 4 mg/L} to highly resistant (MIC >256 mg/L). Emerg Infect Dis, 2004 Sep, 10(9), 1671 - 3 Acinetobacter baumannii in human body louse; La Scola B et al.; While we were isolating Bartonella quintana from body lice, 40 Acinetobacter baumannii strains were also isolated and genotyped . One clone was unique and the other was ampicillin susceptible . A . baumannii DNA was later detected in 21% of 622 lice collected worldwide . These findings show an A . baumannii epidemic in human body lice. J Microbiol Immunol Infect, 2004 Oct, 37(5), 301 - 6 Neonatal sepsis in the neonatal intensive care unit: characteristics of early versus late onset; Jiang JH et al.; Neonatal sepsis is a major cause of death in newborns despite sophisticated neonatal intensive care . This retrospective study reviewed the clinical characteristics of cases of culture-proven sepsis in a neonatal intensive care unit from January 1992 to December 2001 . Patients were divided into those with onset of sepsis in the first 7 days of life (early-onset group) and those with onset after the seventh day of life (late-onset group) . A total of 270 cases with 325 episodes of sepsis and 353 isolated pathogens were identified and included in the study . The male-to-female ratio was 1.4 . The majority of cases of sepsis occurred in low birth weight (75.9%) and premature babies (76.7%) . Late onset occurred in 71.9% of cases . Patients with late onset had a lower mortality rate than those with early onset (11.3% vs 28.9%) . Coagulase-negative staphylococci (20.1%) was the most common organism isolated, but infection with Pseudomonas aeruginosa was associated with the highest morality rate (55.0%) . Late-onset sepsis was significantly more common in very low birth weight and premature infants . The most frequently encountered pathogens in the early-onset group were group B streptococci (GBS) and Escherichia coli, while in the late-onset group, the organisms were coagulase-negative staphylococci and Enterobacteriaceae, including E . coli, Klebsiella pneumoniae, and Acinetobacter baumannii . GBS infection resulted in the highest mortality when the onset of sepsis was within the first 24 hours of life. Semin Pediatr Infect Dis, 2004 Oct, 15(4), 280 - 7 Monkeypox: a review of the history and emergence in the Western hemisphere; Ligon BL; A mysterious disease was reported on May 24, 2003, when the Wisconsin Division of Public Health (DPH) received notice of a 3-year-old girl who had been hospitalized in central Wisconsin with cellulites and fever after being bitten by a prairie dog on May 13 . The laboratory isolated a gram-negative bacillus, raising concerns that it might be tularemia or plague; ultimately, it was identified as an acinetobacter species and was considered to be a contaminant . Because no other such cases were reported at the time, the case was thought to be merely an isolated event . However, within two weeks, on June 2, 2003, evidence of a much wider scenario began to emerge . On that date, the Wisconsin DPH received notice from the Marshfield Laboratory that the mother of the first patient had become ill on May 26 and that electron-microscopic evidence of a poxvirus was found in a skin lesion . On that same day, another report, this time from the Milwaukee Health Department, of a strange illness was received at the DPH and described the case of a meat inspector who resided in southeastern Wisconsin and also was a distributor of exotic animals . By July 30, 2003, 72 confirmed or suspected cases of monkeypox had been reported in Wisconsin, Illinois, and Indiana and represented a large outbreak . The peak in the onset of illness occurred between May 29 and June 9, 2003, and no further cases of illness have been reported in humans since June 22, 2003 . Traceback investigations from the child and other patients followed the route of introduction of monkeypox into Wisconsin to a distributor in Illinois, who had received a shipment of exotic animals imported into the United States through Texas from Ghana, West Africa. Saudi Med J, 2004 Oct, 25(10), 1403 - 9 Investigation of the antibiotic susceptibility patterns of pathogens causing nosocomial infections; Yaman A et al.; OBJECTIVE: The aim of this study is to determine the resistance patterns of bacteria causing nosocomial infections . The outcome of this resistance was followed for 3 years . METHODS: This study was carried out during 2000 to 2002 at a university hospital in Turkey . The resistance patterns of 570 bacteria (390 Gram-negative, 180 Gram-positive) against meropenem, imipenem, ceftazidime, cefotaxime, cefepime, piperacillin/tazobactam, ciprofloxacin and tobramycin were investigated using the E-test . Extended-spectrum beta-lactamase (ESBL) production was determined using ceftazidime and ceftazidime/clavulanic acid E-test strips . RESULTS: Meropenem was the most effective antibiotic against Gram-negative organisms (89.0%); this was followed by imipenem (87.2%) and piperacillin/tazobactam (66.4%) . The most active antibiotic against Gram-positive bacteria was imipenem (87.2%) and this was followed by piperacillin/tazobactam (81.7%) and meropenem (77.8%) . The rates of production of ESBL by Escherichia coli were 20.9%, Klebsiella pneumoniae 50% and Serratia marcescens were 46.7% . Extended-spectrum beta-lactamase production increased each year (21.7%, 22.1% and 45.5%) . All of the ESBL producing isolates were sensitive to meropenem and 98.5% sensitive to imipenem . AmpC beta-lactamase was produced by 20.9% of the Enterobacter species spp, Citrobacter spp . and Serratia marcescens . All of these were sensitive to meropenem and 77.8% to imipenem and ciprofloxacin . Multi-drug resistance rates in Acinetobacter spp were 45.4% and 37.7% in Pseudomonas aeruginosa isolates . CONCLUSION: As in the entire world, resistance to antibiotics is a serious problem in our country . Solving of this problem depends primarily on prevention of the development of resistance. Oral Microbiol Immunol, 2004 Dec, 19(6), 363 - 70 Investigation of bacterial communities associated with asymptomatic and symptomatic endodontic infections by denaturing gradient gel electrophoresis fingerprinting approach; Siqueira JF Jr et al.; The purpose of the present study was to investigate the bacterial communities associated with asymptomatic and symptomatic endodontic infections and to compare denaturing gradient gel electrophoresis (DGGE) fingerprinting patterns of these two clinical conditions . The root canal microbiota of teeth associated with asymptomatic or symptomatic periradicular lesions was profiled by the PCR-DGGE method and then compared, taking into consideration the banding patterns . Bacteria were present in all examined cases . Comparative analysis of the two clinical conditions revealed bands that were common to both symptomatic and asymptomatic cases, but most DGGE bands appeared to be unique for each clinical condition . No single band occurred in all profiles . The mean number of bands detected in the 16S rDNA community profiles were 12.1 +/- 9.4 (range 2-29) for symptomatic samples and 6.7 +/- 2.7 (range 2-11) for asymptomatic ones . Clustering methods and principal component analysis of DGGE banding pattern placed the samples according to the presence or absence of symptoms . Four intense bands that were excised from the gel and sequenced showed similarities to species of the Campylobacter genus (found in 5/12 asymptomatic and in 3/11 symptomatic cases), Fusobacterium genus (4/11 symptomatic cases), Acinetobacter genus (5/12 asymptomatic cases), and Enterobacteriaceae family (11/12 asymptomatic and 2/11 symptomatic cases) . The profiles of the predominant bacterial community appeared to be unique for each individual . These findings confirm that endodontic infections are polymicrobial and showed that there are significant differences in the predominant bacterial composition between asymptomatic and symptomatic cases. Arch Microbiol, 2004 Dec, 182(6), 458 - 466 Epub 2004 Oct 7. Relations and functions of dye-linked formaldehyde dehydrogenase from Hyphomicrobium zavarzinii revealed by sequence determination and analysis; Schwartz AC et al.; faoA, the gene of the dye-linked NAD(P)-independent quinone-containing formaldehyde dehydrogenase of methylamine-grown Hyphomicrobium zavarzinii strain ZV 580 was sequenced and analyzed together with an apparent promoter region and adjoining genes in a 7.2-kb fragment of hyphomicrobial DNA . The formaldehyde dehydrogenase, identified as a periplasmic enzyme by its signal sequence, is distantly related to the soluble pyrroloquinoline-quinone-dependent glucose dehydrogenase of Acinetobacter calcoaceticus and to other predicted glucose dehydrogenase sequences . The promoter region, containing about 400 nucleotides upstream of faoA, comprised potential binding sites identical or highly similar to known consensus sequences of the sigma factors sigma(70) (housekeeping), sigma(H) (heat shock), sigma(F) (flagellar) and sigma(N) (nitrogen) . The complex regulation of the transcription of faoA, which is suggested by this setting and emphasized by a possible heat-shock promoter, supports a hypothesis proposing an auxiliary role of the enzyme in lowering detrimental elevated concentrations of formaldehyde, which might arise in the course of stress or regulatory transitions disturbing balanced C(1) metabolism. Braz J Infect Dis, 2004 Jun, 8(3), 206 - 10 Epub 2004 Sep 29. Hospital gowns as a vehicle for bacterial dissemination in an intensive care unit; Pilonetto M et al.; The microbiota from the uniforms of 31 professionals from the general intensive care unit was analyzed . The samples were collected in duplicate at the beginning and at the end of the work period . Total viable counts of microorganisms were determined; there was a significant increase in the counts at the end of the period, when compared with those obtained at the beginning . No significant difference was observed between the first and second counts obtained from the cuffs . However, differences were observed for the samples from the abdominal region . Among the isolated pathogens 11/18 were Staphylococcus aureus, 2/18 were Acinetobacter baumannii, 2/18 were Klebsiela pneumoniae and 1/18 were Serratia rubidae . Some of these isolates were multi-resistant to antibiotics . Emphasis should be placed on reducing the spread of these pathogens in the hospital, making sure that biosafety protocols are followed by the staff. Braz J Infect Dis, 2004 Jun, 8(3), 190 - 6 Epub 2004 Sep 29. Use of selective media for detection of cephalothin-resistant bacteria in surgical patients; Melo SA et al.; Bacterial colonization in hospitalized patients is an important step in nosocomial infections . Frequent employment of antimicrobials can modify patients' normal microbiota, favoring colonization and infection by antimicrobial-resistant microorganisms . First-generation cephalosporins are frequently used as prophylactic antibiotics in surgery . Intestinal, oropharyngeal and skin colonization by cephalothin-resistant microorganisms were studied in 60 pre-operative patients at the Hospital Universitario Pedro Ernesto . Feces were cultured in Eosin-methylene blue medium containing 32 microg/mL of cephalothin . Swabs obtained from the oropharynx and from skin were inoculated in cistein-lactose electrolytes-deficient medium containing 32 microg/mL of cephalothin . Isolated strains were identified and tested for susceptibility to antimicrobials by disk diffusion . Cephalothin-resistant strains were isolated from the feces of 59 patients (98%), from the oropharynx of 13 patients (22%) and from skin in 10 patients (17%) . Enterobacter cloacae was predominant in feces (68% of the patients) and oropharynx (13%) . Acinetobacter spp . was the most frequent microorganism isolated from the skin (10%) . Antimicrobial multiresistant strains were isolated from at least one of the sites in 38 patients (63%) . The employment of selective medium containing antimicrobials is a relatively simple and efficient method, being useful to evaluate microorganisms from hospitalized patients' microbiota that are relevant as potential pathogens in nosocomial infections. J Clin Microbiol, 2004 Oct, 42(10), 4776 - 9 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; Lee SO et al.; 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. J Antimicrob Chemother, 2004 Nov, 54(5), 870 - 875 Epub 2004 Oct 7. Evaluation of the MicroScan ESBL plus confirmation panel for detection of extended-spectrum {beta}-lactamases in clinical isolates of oxyimino-cephalosporin-resistant Gram-negative bacteria; Sturenburg E et al.; OBJECTIVE: We aimed to assess the performance of the MicroScan ESBL plus confirmation panel using a series of 87 oxyimino-cephalosporin-resistant Gram-negative bacilli of various species . METHODS: Organisms tested included 57 extended-spectrum beta-lactamase (ESBL) strains comprising Enterobacter aerogenes (3), Enterobacter cloacae (10), Escherichia coli (11), Klebsiella pneumoniae (26), Klebsiella oxytoca (3) and Proteus mirabilis (4) . Also included were 30 strains resistant to oxyimino cephalosporins but lacking ESBLs, which were characterized with other resistance mechanisms, such as inherent clavulanate susceptibility in Acinetobacter spp . (4), hyperproduction of AmpC enzyme in Citrobacter freundii (2), E . aerogenes (3), E . cloacae (3), E . coli (4), Hafnia alvei (1) and Morganella morganii (1), production of plasmid-mediated AmpC beta-lactamase in K . pneumoniae (3) and E . coli (3) or hyperproduction of K1 enzyme in K . oxytoca (6) . RESULTS: The MicroScan MIC-based clavulanate synergy correctly classified 50 of 57 ESBL strains as ESBL-positive and 23 of 30 non-ESBL strains as ESBL-negative (yielding a sensitivity of 88% and a specificity of 76.7%, respectively) . False negatives among ESBL producers were highest with Enterobacter spp . due to masking interactions between ESBL and AmpC beta-lactamases . False-positive classifications occurred in two Acinetobacter spp., one E . coli producing plasmid-mediated AmpC beta-lactamase and two K . oxytoca hyperproducing their chromosomal K1 beta-lactamase . CONCLUSION: The MicroScan clavulanate synergy test proved to be a valuable tool for ESBL confirmation . However, this test has limitations in detecting ESBLs in Enterobacter spp . and in discriminating ESBL-related resistance from the K1 enzyme and from inherent clavulanate susceptibility in Acinetobacter spp. Rev Esp Quimioter, 2004 Jun, 17(2), 177 - 83 {Epidemiological study and effect on antimicrobial use in the genus Acinetobacter in a university hospital}; Iglesias de Sena H et al.; In recent years, bacteria of the genus Acinetobacter have gradually been gaining epidemiological importance . This is due to the fact that they have been emerging as opportunistic pathogens causing a great number of nosocomial infections, and due to their multiresistance to antimicrobial agents, which limits treatment options . This study aimed to determine the epidemiological importance of isolates of the genus Acinetobacter at the Clinical University Hospital of Salamanca, Spain . The identification and susceptibility of Acinetobacter was determined using the Pasco automated system for Gram-negative bacteria MIC/ID 6J . Clinical and epidemiological data were obtained from the clinical history . During the study period it was found that A . anitratus was the most frequently isolated species (56%) . The origin of the bacteria was mainly nosocomial (70%), and it was most prevalent in the ICU. Arch Biochem Biophys, 2004 Nov 1, 431(1), 79 - 87 Structural roles of the active site iron(III) ions in catechol 1,2-dioxygenases and differential secondary structure changes in isoenzymes A and B from Acinetobacter radioresistens S13; Di Nardo G et al.; The reversible active site metal ion removal process for two catechol 1,2-dioxygenase isoenzymes (IsoA and IsoB) isolated from Acinetobacter radioresistens S13 has been monitored using circular dichroism and fluorescence spectroscopic techniques . IsoA and IsoB are homodimers, containing one iron(III) ion per subunit . Their amino acid sequence identity is 48.4% . Previous experiments suggested that structural diversities could be responsible for the differential thermal and pH stabilities of the two isoenzymes and of their distinct demetallation kinetics . The far-UV CD spectra of IsoA and IsoB catechol 1,2-dioxygenases from A . radioresistens S13 provide information on their secondary structures . IsoB appears to have a content of alpha-helices higher than IsoA . Upon metal ion removal, both proteins reversibly lose part of their secondary structure following distinct pathways . CD spectra simulations allowed us to estimate the content of alpha-helices, beta-sheets, and turns for each isoenzyme and to monitor the secondary structure rearrangements . The metal ion withdrawal has large influence on the secondary structure: in particular a significant reduction of alpha-helices content is observed for both isoenzymes . Intrinsic fluorescence emission spectra clearly support such results, adding information on the local environment changes of the tryptophan residues . The positioning of Trp250 in IsoB has been shown to be of particular interest for monitoring the local structure changes occurring upon metal ion removal . For the first time these studies allow to underline the role of active site iron ions on dioxygenases folding and stability, further evidencing the differences in structural assembling between the two isoenzymes from A . radioresistens S13. Expert Opin Investig Drugs, 2004 Oct, 13(10), 1307 - 18 Beta-lactamase inhibitors: evolving compounds for evolving resistance targets; Georgopapadakou NH; The many and diverse beta-lactamases produced by bacteria, particularly by Gram-negative pathogens, are increasingly posing a serious threat to the clinical utility of beta-lactams . First-generation inhibitors (clavulanic acid, sulbactam, tazobactam) focus on Ambler class A enzymes . However, recent structural upgrades of class A beta-lactamases (e.g . TEM, SHV) have extended their spectrum (extended-spectrum beta-lactamases and carbapenemases {Sme, NMC-A, IMI-1}) and have brought about the possibility of beta-lactamase-inhibitor resistance . Furthermore, the mobilisation and spread of originally chromosomal class C enzymes (CMY, MIR), the growing clinical importance of class B enzymes (IMP, VIM), the emergence of inhibitor-resistant, broad spectrum class D (OXA) enzymes and the co-existence of different classes of beta-lactamases in the same pathogen have spurred research toward universal inhibitors . A complicating issue is target accessibility in Gram-negative bacteria, particularly in Enterobacter, Acinetobacter, Pseudomonas, Stenotrophomonas and other organisms, which is necessary in order for the inhibitor to synergise with vulnerable beta-lactam antibiotics . Several new, broad-spectrum inhibitors have emerged: cephem sulfones and oxapenems are upgrades of penam sulfones and oxapenams, respectively, with cephem sulfones possibly extending their inhibition to class B metallo-enzymes; and boronates and phosphonates are designed de novo, based on common structural and mechanistic features of serine beta-lactamases. Am J Infect Control, 2004 Oct, 32(6), 342 - 4 Direct costs of multidrug-resistant Acinetobacter baumannii in the burn unit of a public teaching hospital; Wilson SJ et al.; We conducted a case-control study to determine the attributable direct costs of multidrug-resistant Acinetobacter baumannii (MDRAB) in the burn unit of a public teaching hospital . The mean total hospital cost of patients who acquired MDRAB was 98,575 dollars higher than that of control patients who had identical burn severity of illness indices ( P <.01) . These data should help infection control practitioners and others determine the cost-effectiveness of specific interventions designed to control this emerging nosocomial pathogen. Chemotherapy, 2004 Oct, 50(4), 202 - 10 Epub 2004 Sep 23. Activity of levofloxacin and ciprofloxacin in combination with cefepime, ceftazidime, imipenem, piperacillin-tazobactam and amikacin against different Pseudomonas aeruginosa phenotypes and Acinetobacter spp; Drago L et al.; BACKGROUND: Combination therapy is used to widen the antimicrobial spectrum, minimize toxicity and prevent the emergence of resistant mutants . METHODS: Synergy between levofloxacin or ciprofloxacin and ceftazidime, cefepime, imipenem, piperacillin-tazobactam and amikacin was evaluated by checkerboard assay with 55 strains and by time-kill curves with 8 strains of Pseudomonas aeruginosa and Acinetobacter spp . RESULTS: In the checkerboard assay, synergy and additivity were the most frequent effects observed among all the combinations against P . aeruginosa and Acinetobacter spp., with no significant differences between the two fluoroquinolones . No antagonism was observed . In the time-kill curves, synergy was evidenced against all the tested strains, at least for one combination at one of the time points considered . Levofloxacin and ciprofloxacin combined with ceftazidime, as well as levofloxacin plus amikacin, were synergistic for all the strains tested . CONCLUSION: Combinations of fluoroquinolones with beta-lactams or amikacin show an enhanced activity against P . aeruginosa and Acinetobacter spp. Biochim Biophys Acta, 2004 Oct 5, 1680(1), 60 - 6 Cloning and expression of p-hydroxyphenylacetate 3-hydroxylase from Acinetobacter baumannii: evidence of the divergence of enzymes in the class of two-protein component aromatic hydroxylases; Thotsaporn K et al.; The genes encoding for the reductase and oxygenase components of p-hydroxyphenylacetate 3-hydroxylase from Acinetobacter baumannii were cloned and expressed in an E . coli system . The recombinant enzymes were purified and shown to have the same catalytic properties as the native enzyme . Sequence analysis and biochemical studies indicate that the enzyme represents a novel prototype of enzyme in the two-protein component class of aromatic hydroxylases . The C2 component shows little similarity to other oxygenases in the same class, correlating with its uniquely broad flavin specificity . Analysis of the C1 reductase sequence indicates that the binding sites of flavin and NADH mainly reside in the N-terminal half while the C-terminal half may be responsible for HPA-stimulation of NADH oxidation. RNA, 2004 Nov,