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Characterization of the aac(6')-Ik gene of Acinetobacter sp. 6.

The aac(6')-Ik gene which confers resistance to aminoglycosides in Acinetobacter sp. 6 CIP A165 was characterized. The resistance gene was identified as a coding sequence of 438 bases pairs corresponding to a protein with a calculated mass of 16627 Da. Alignment of aac(6')-Ik with aac(6')-Ig from Acinetobacter haemolyticus indicated 83% identity. Like aac(6')-Ig of A. haemolyticus and aac(6')-Ij of Acinetobacter sp. 13, the aac(6')-Ik gene was apparently located in the chromosome and was species specific. The high degree of identity between aac(6')-Ig and -Ik, compared with genomic DNA relationships between the host species, indicated that these genes have diverged from a common ancestor in a parental Acinetobacter species.

Acetyltransferases↗

Lomefloxacin (SC 47111 or NY-198), a new difluorinated quinolone: comparison of the in vitro activity with other broad spectrum antimicrobials against Enterobacteriaceae, Acinetobacter spp, Aeromonas spp, and Pseudomonas aeruginosa.

Using a broth microdilution method, we compared the in vitro activity of lomefloxacin to other broad spectrum antimicrobials against clinical strains of Enterobacteriaceae, Acinetobacter spp, Aeromonas spp, and P. aeruginosa. Against the Enterobacteriaceae and A. hydrophila, lomefloxacin showed excellent activity with MIC50 values ranging from 0.12-0.5 micrograms/ml and MIC90 values ranging from 0.12-1.0 micrograms/ml. By comparison, lomefloxacin had superior activity to nalidixic acid and difloxacin, comparable activity to norfloxacin and fleroxacin, and slightly less activity than ciprofloxacin. Lomefloxacin was also more active than imipenem, ceftazidime, aztreonam, ticarcillin/clavulanic acid (T-CA), and gentamicin. Lomefloxacin was less active (MIC90, 4 micrograms/ml) against Acinetobacter strains than Enterobacteriaceae strains. Against these same Acinetobacter strains, lomefloxacin was 8- to 64-fold more active than ceftazidime, aztreonam, T-CA, and gentamicin but slightly less active than imipenem. Lomefloxacin was 2- to 8-fold more active than fleroxacin, difloxacin, and nalidixic acid against strains of P. aeruginosa. However, lomefloxacin was less active than ciprofloxacin and norfloxacin. Lomefloxacin had comparable activity to imipenem but was 2- to 16-fold more active than ceftazidime, aztreonam, T-CA, and gentamicin. Thus, lomefloxacin demonstrated comparable activity to other fluorinated quinolones in that it possesses high active against clinical strains of Enterobacteriaceae and had good-to-moderate activity against most strains of Acinetobacter and P. aeruginosa.

4-Quinolones↗

In vitro activity of beta-lactamase inhibitors against clinical isolates of Acinetobacter species.

Acinetobacter is an important cause of nosocomial infections, and it is often resistant to many antibiotics. In a search for alternative agents, three beta-lactamase inhibitors (sulbactam, clavulanate, and tazobactam) and five beta-lactam antibiotics (imipenem, ceftazidime, ceftriaxone, cefotaxime, and piperacillin) were tested against 68 unique clinical isolates of Acinetobacter species. Minimum inhibitory concentrations were determined by a broth microdilution method. Using temperature sensitivity testing, we identified 59 strains as Acinetobacter baumannii, one as Acinetobacter haemolyticus, and eight as indeterminate biotype species. We demonstrated 41 of 59 (70%) strains of A. baumannii to be multiply resistant (susceptible only to amikacin and imipenem), whereas all the nonbaumannii strains were not. Imipenem was the most active agent among the compounds investigated. All three beta-lactamase inhibitors had strong intrinsic activity, with sulbactam being the most active agent among the beta-lactamase inhibitors studied.

Acinetobacter↗

bla(OXA-51)-type beta-lactamase genes are ubiquitous and vary within a strain in Acinetobacter baumannii.

We investigated the spread of bla(OXA-51)-type beta-lactamase genes in 200 Acinetobacter spp. clinical strains isolated in Argentina from 1982 to 2005. bla(OXA-51)-type genes were present in all Acinetobacter baumannii isolates tested (n=194), whereas they were not detected in two Acinetobacter haemolyticus, two genomic species 10 or two Acinetobacter lwoffii isolates. The bla(OXA-51)-type alleles varied within a strain and were found in six different A. baumannii pulsed-field gel electrophoresis clones that were susceptible or resistant to imipenem, suggesting a controversial role in imipenem resistance. Our findings agree with previous reports showing that bla(OXA-51)-type genes are naturally harboured by A. baumannii isolates from various geographical origins and support the presence of a direct reservoir of beta-lactam resistance genes within the nosocomial environment.

Acinetobacter baumannii↗

Influence of phenolics on the sensitivity of free and immobilized bioluminescent Acinetobacter bacterium.

In this work, the constructed bioluminescent Acinetobacter strain DF4/PUTK2 was employed to assess the toxicity of phenolic compounds and the 5 min EC50 values were calculated. The results of the DF4/PUTK2 assay were further evaluated by comparing with the results of the Vibrio fischeri luminescence inhibition assay. To develop a bioassay system appropriate to be used in continuous toxicity testing, strain DF4/PUTK2 was subjected for immobilization in microtiter plates into the matrices Ca-alginate, polyacrylamide, agar and agarose. After a choice of materials was tried, Ca-alginate was selected as the most suitable candidate material. Because, it could be stored at least 8 weeks at 4 degrees C, during which the ability of the bioreporter DF4/PUTK2 to detect the toxicity of phenolics was maintained. However, the stability of the bioluminescence for DF4/PUTK2 cells immobilized into agarose and agar was significantly less than that of cells stored in alginate suspensions. This study recommended that luxCDABE-marked Acinetobacter strain DF4/PUTK2 could be employed to assay the ecotoxicity of environmental samples contaminated with phenols. The host strain of the bioreporter DF4/PUTK2 is Acinetobacter strain DF4. It is known that members of the genus Acinetobacter are widespread in nature and also involved in biodegradation, leaching and removal of several organic and inorganic man-made hazardous wastes.

Acinetobacter↗

Estimation of ribosomal RNA operon (rrn) copy number in Acinetobacter isolates and potential of patterns of rrn operon-containing fragments for typing strains of members of this genus.

The copy number of the rrn operon in 70 strains of Acinetobacter including the type strains of almost all the genomic species with validated names was estimated after digestion of their genomic DNA by the restriction enzymes BglII and PstI, and Southern blotting. Copy number estimates varied between and among species, with between 3 and 7 rrn operon copies detected. Copy number estimates obtained from the same strain with the two enzymes sometimes varied. BglII generated RFLP patterns of the rrn containing fragments obtained from Southern blots after agarose gel electrophoresis were examined for their value in identifying Acinetobacter isolates. This method was very reproducible with the same fragment pattern always generated from the same isolate on repeated analysis. Often multiple strains of the same genomic species gave identical or very similar patterns (e.g. Acinetobacter baylyi), clustering closest together on the dendrogram generated after numerical analysis of these patterns. However, with some, like BG5 and BG8, the patterns derived from the different strains, some of which had been placed in this genomic species from DNA:DNA hybridization data, varied considerably to each other and to the type strain. Little similarity was seen when relationships between these strains based on these patterns were compared to those using DNA:DNA hybridization data. Often these patterns could be used to question earlier identification of strains using phenotypic characters. Thus, strain AB82 thought to belong to genomic species 5 gave an identical pattern to A. bouvetii(T) (DSM 14964). In some cases this pattern analysis suggested that novel species of Acinetobacter might exist among the strains examined.

Acinetobacter↗

Nitrite inhibition of aerobic growth of Acinetobacter sp.

Nitrite inhibition of Acinetobacter sp. growing under aerobic conditions was studied. Specific growth rates under non-limiting concentrations of acetate and dissolved oxygen averaged 0.62h(-1). Growth and phosphate uptake by Acinetobacter sp. were both inhibited by increasing nitrite concentrations. The median inhibitory concentration (IC50) of free nitrous acid (FNA) was 0.10 mg/L and the IC10 of FNA was 0.05 mg/L. Removing nitrite from cultures reversed the inhibitory effect. Comparison of the IC10 of FNA for Acinetobacter sp. to inhibitory concentrations for other wastewater heterotrophic bacteria suggests that Acinetobacter sp. are relatively sensitive to this compound.

Acinetobacter↗

Cross-reactivity between related sequences found in Acinetobacter sp., Pseudomonas aeruginosa, myelin basic protein and myelin oligodendrocyte glycoprotein in multiple sclerosis.

To investigate the possible role of molecular mimicry to bacterial components in multiple sclerosis (MS) pathogenesis we examined antibody responses to mimicry peptide sequences of Acinetobacter, Pseudomonas aeruginosa and myelin components. Antibodies to mimicry peptides from Acinetobacter (p<0.001), P. aeruginosa (p<0.001), myelin basic protein (MBP) (p<0.001) and myelin oligodendrocyte glycoprotein (MOG) (p<0.001) were significantly elevated in MS patients compared to controls. Antisera against MBP (residues 110-124) reacted with both Acinetobacter and Pseudomonas peptides from 4- and gamma-carboxymuconolactone decarboxylase, respectively. MOG (residues 43-57) antisera reacted with Acinetobacter peptide from 3-oxo-adipate-CoA-transferase subunit A. The role of these bacteria in MS is unclear but demonstrates that molecular mimicry is not restricted to viruses suggesting bacterial infections could play a role in MS pathogenesis. Further work is required to evaluate the relevance of these cross-reactive antibodies to the neuropathology of MS.

Acinetobacter↗

Acinetobacter bacteremia in patients with diarrhoeal disease.

In 1994, 171 (27%) of all positive blood cultures in our hospital were due to Acinetobacter species. Of these, 138 cultures were considered significant, 91 (66%) were community-acquired and 47 (34%) were nosocomial. Most acinetobacter bacteraemia in children < or = 1 year old was community-acquired, while nosocomial infection was more common in children > 1 year old (P = 0.01). Most children < or = 5 years old were severely malnourished. The incidence of bacteraemia was lowest during the post-monsoon to early winter months. Acinetobacter bacteraemia associated mortality was twice (16%) that of all other patients (7.7%, P < 0.0005) and accounted for 4.5% of all hospital deaths during the study period. Bacteraemia caused by Acinetobacter species is an important cause of morbidity and mortality among our patient population with diarrhoeal disease.

Acinetobacter Infections↗

Phenotypic characterization and antibiotic resistance of Acinetobacter spp. isolated from aquatic sources.

A total of 99 Acinetobacter isolates from sewage, freshwater aquaculture habitats, trout intestinal contents and frozen shrimps was characterized phenotypically and antibiotic susceptibility patterns determined. One group of genomic species, including Ac. johnsonii, Ac. lwoffi and spp. 15TU, was detected in all sample types and represented the majority of the isolates (n = 54). Isolates belonging to the Acb complex (Ac. calcoaceticus, Ac. baumannii and genomic species 3) were detected in sewage (n = 6) and frozen shrimps (n = 1), Ac. haemolyticus in frozen shrimps (n = 6) and trout intestinal contents (n = 2) and genomic species 11 in freshwater aquaculture habitats (n = 6) and trout intestinal contents (n = 1). Acinetobacter junii (n = 5), genomic species 10 (n = 2), 14BJ (n = 8) and 16BJ (n = 4) were only isolated from sewage. Acinetobacter isolates from sewage were generally more biochemically reactive and resistant to antimicrobial agents compared with isolates from other sample types. Different strains, often belonging to different genomic species, were isolated from sites situated upstream and downstream of the discharge point of a pharmaceutical plant. This finding supported the hypothesis that the waste effluent from the pharmaceutical plant was likely to cause a change in the distribution of Acinetobacter spp. by selecting and/or introducing antibiotic-resistant strains into the recipient sewers.

Acinetobacter↗

The rapid identification of Acinetobacter species using Fourier transform infrared spectroscopy.

AIMS: Fourier transform infrared (FT-IR) was used to analyse a selection of Acinetobacter isolates in order to determine if this approach could discriminate readily between the known genomic species of this genus and environmental isolates from activated sludge. METHODS AND RESULTS: FT-IR spectroscopy is a rapid whole-organism fingerprinting method, typically taking only 10 s per sample, and generates 'holistic' biochemical profiles (or 'fingerprints') from biological materials. The cluster analysis produced by FT-IR was compared with previous polyphasic taxonomic studies on these isolates and with 16S-23S rDNA intergenic spacer region (ISR) fingerprinting presented in this paper. FT-IR and 16S-23S rDNA ISR analyses together indicate that some of the Acinetobacter genomic species are particularly heterogeneous and poorly defined, making characterization of the unknown environmental isolates with the genomic species difficult. CONCLUSIONS: Whilst the characterization of the isolates from activated sludge revealed by FT-IR and 16S-23S rDNA ISR were not directly comparable, the dendrogram produced from FT-IR data did correlate well with the outcomes of the other polyphasic taxonomic work. SIGNIFICANCE AND IMPACT OF THE STUDY: We believe it would be advantageous to pursue this approach further and establish a comprehensive database of taxonomically well-defined Acinetobacter species to aid the identification of unknown strains. In this instance, FT-IR may provide the rapid identification method eagerly sought for the routine identification of Acinetobacter isolates from a wide range of environmental sources.

Acinetobacter↗

Seasonal variation of Acinetobacter infections: 1987-1996. Nosocomial Infections Surveillance System.

To determine whether nosocomial infections due to Acinetobacter species have increased over the past 10 years and whether infections continue to have a pronounced seasonal variation, we analyzed infections reported by hospitals in the National Nosocomial Infections Surveillance System that performed adult and pediatric intensive care unit surveillance from 1987 through 1996. Overall, 3447 nosocomial acinetobacter infections were reported during 5,596, 156 patient-days. There was a yearly median of 7.2 infections (range, 5.0-10.5) per 10,000 patient-days and a downward trend in the rate of acinetobacter infections overall (P<.05) and of 2 major types of infection (P<.05): bloodstream infections (yearly median, 1.6 per 10, 000 central venous catheter-days; range, 1.3-2.9) and pneumonia (yearly median, 7.6 per 10,000 ventilator-days; range, 6.5-12.0). Throughout this period, average rates were significantly higher during July-October than during November-June for acinetobacter infections overall (8.0 vs. 5.2; P<.01) and for bloodstream infections (2.0 vs. 1.2; P<.01) and pneumonia (9.7 vs. 6.6; P<.01).

Acinetobacter Infections↗

Comparison of ampicillin-sulbactam and imipenem-cilastatin for the treatment of acinetobacter ventilator-associated pneumonia.

Acinetobacter organisms, which are a common cause of ventilator-associated pneumonia (VAP) in some health care centers, are becoming more resistant to such first-line agents as imipenem-cilastatin (Imi-Cil). Sulbactam has good in vitro activity against Acinetobacter organisms; thus, ampicillin-sulbactam (Amp-Sulb) may be a viable treatment alternative. The outcomes for critically ill trauma patients with Acinetobacter VAP treated with either Amp-Sulb or Imi-Cil were compared retrospectively. A total of 77 episodes in 75 patients were studied. Fourteen patients were treated with Amp-Sulb, and 63 patients were treated with Imi-Cil. Treatment efficacy was similar in the Amp-Sulb and Imi-Cil groups (93% vs. 83%, respectively; P>.05). No statistically significant differences between groups were noted with regard to associated mortality, duration of mechanical ventilation, or length of stay in the intensive care unit or hospital. However, adjunctive aminoglycoside therapy was used more often in the Amp-Sulb group. Patients generally received Amp-Sulb because of imipenem resistance. Amp-Sulb was effective in treating a small number of patients with Acinetobacter VAP; however, more data are needed.

Acinetobacter↗

Emergence of resistance in Pseudomonas aeruginosa and Acinetobacter species after the use of antimicrobials for burned patients.

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.

Acinetobacter↗

In vitro selection of resistance in Pseudomonas aeruginosa and Acinetobacter spp. by levofloxacin and ciprofloxacin alone and in combination with beta-lactams and amikacin.

OBJECTIVES: The aim of this study was to evaluate the ability of levofloxacin and ciprofloxacin alone and in combination with either ceftazidime, cefepime, imipenem, piperacillin-tazobactam or amikacin to select for antibiotic-resistant mutants of Pseudomonas aeruginosa and Acinetobacter spp. METHODS: Clinical strains of P. aeruginosa (n = 5) and Acinetobacter spp. (n = 5) susceptible to all the drugs used in the study were assayed. Development of resistance was determined by multi-step and single-step methodologies. For multi-step studies, MICs were determined after five serial passages on antibiotic-gradient plates containing each antibiotic alone or in combination with levofloxacin or ciprofloxacin. Acquisition of resistance was defined as an increase of >or=4-fold from the starting MIC. In single-step studies, the frequency of spontaneous mutations was calculated after a passage on plates containing antibiotics alone and in combinations at concentrations equal to the highest NCCLS breakpoints. RESULTS: Serial passages on medium containing single antibiotics resulted in increased MICs for each antibiotic; MIC increases were limited by antibiotics in combination. A decrease in the number of strains with MICs above the NCCLS breakpoints occurred when fluoroquinolones were combined with a second antibiotic for both P. aeruginosa and Acinetobacter spp. isolates. Frequencies of mutation were higher for antibiotics alone than for combinations. CONCLUSIONS: Use of combinations of fluoroquinolones with beta-lactams and amikacin reduces the risk for in vitro selection of resistant P. aeruginosa and Acinetobacter spp.

Acinetobacter↗

Acinetobacter parvus sp. nov., a small-colony-forming species isolated from human clinical specimens.

The taxonomic status of seven glucose-non-acidifying, non-proteolytic Acinetobacter strains characterized by forming small colonies on agar media was studied. With one exception, all strains were from human specimens. They could be distinguished from all described Acinetobacter (genomic) species by their ability to grow on ethanol and acetate as sole sources of carbon but not on 22 other substrates tested including DL-lactate or DL-4-aminobutyrate. DNA-DNA hybridization studies, 16S rRNA gene sequence analysis, amplified rDNA restriction analysis and DNA polymorphism analysis by AFLP showed that these strains represent a hitherto unknown species of the genus Acinetobacter, for which the name Acinetobacter parvus (type strain LMG 21765(T)=LUH 4616(T)=NIPH 384(T)=CCM 7030(T)) is proposed.

Acinetobacter↗

The metabolism of trans-cyclohexan-1,2-diol by an Acinetobacter species.

1. Acinetobacter TD63 was one of some thirty organisms isolated by elective culture with trans-cyclohexan-1,2-diol as sole source of carbon. The great majority of these isolates displayed the same growth spectrum as Nocardia globerula CL1 and Acinetobacter NCIB 9871 being capable of utilizing trans-cyclohexan-1,2-diol, 2-hydroxycyclohexan-1-one, cyclohexanol, cyclohexanone,1-oxa-2oxocycloheptane and adipate and were assumed to use well described metabolic pathways. 2. Acinetobacter TD63 was distinctive in being incapable of growth with cyclohexanol, cyclohexanone or 1-oxa-2-oxocycloheptane and because of this it was hoped that it would display an alternative pathway for the oxidation of trans-cyclohexan-1,2-diol. 3. Studies with cell extracts have shown the presence of inducible dehydrogenase for the conversion of trans-cyclohexan-1,2-diol to 2-hydroxycyclohexan-1-one and cyclohexan-1,2-dione and of 6-oxohexanoate to adipate. These enzymes are linked into a metabolic sequence by the action of a monooxygenase of broad specificity but efficiently capable of converting 2-hydroxy-cyclohexan-1-one into the lactone 1-oxa-2-oxo-7-hydroxycycloheptane that spontaneously rearranges to yield 6-oxohexanoate. 4. An enzyme capable of attacking cyclohexan-1,2-dione (mono-enol) in the absence of an electron donor or oxygen has also been detected. Evidence has been presented indicating that this enzyme catalyses a keto-enol tautomerization between cyclohexan-1,2-dione (mono-enol) and cyclohexan-1,2-dione (mono-hydrate) and is not involved in the pathway of ring cleavage. 5. The failure of Acinetobacter TD63 to grow with cyclohexanol, cyclohexanone or 1-oxa-2-oxocycloheptane is due not to this organism possessing a distinctive metabolic sequence but to a narrow inducer specificity coupled with an inability to form a lactone hydrolase enabling it to cleave the stable 1-oxa-2-oxocycloheptane which is an intermediate in the established pathway of cyclohexanol and cyclohexanone oxidation.

Acinetobacter↗

The phylogenetic structure of the genus Acinetobacter.

16S rDNA sequence analysis was performed on the type strains of all validly described Acinetobacter species and five unnamed Acinetobacter strains. The phylogenetic analyses confirm that Acinetobacter is a coherent genus within the gamma subclass of Proteobacteria and that the species are phylogenetically well defined. A. calcoaceticus, A. lwoffii, A. johnsonii and A. haemolyticus form one cluster of closely related species, the pair A. junii and A. baumannii forms a second cluster. A. radioresistens stands phylogenetically isolated. The study reveals that three undescribed strains can be assigned to individually described species, while strains DSM 30009 and DSM 590 may represent two novel Acinetobacter species.

Acinetobacter↗