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[Degradation of oil derivatives by Acinetobacter calcoaceticus MM5].

This paper describes the isolation of microorganisms from polluted heating oil. The growth of one of them has been studied (Acinetobacter calcoaceticus MM5) in several linear and branched hydrocarbons as well as the effect of its growth on commercial diesel oil. Acinetobacter calcoaceticus MM5 is not capable of using glucose as its only source of carbon, and it needs the presence of nitrogen and phosphorus sources to degrade any petroleum by-product.

Acinetobacter calcoaceticus↗

[Hemolytic properties of bacteria from the genus Acinetobacter].

Hemolytic activity of 370 strains of Acinetobacter was investigated. They represented species A. baumanii (195), A. junii (70), A. lwofii (71) and A. haemolyticus (27) and were isolated from hospitalized patients. Direct and intermediate hemolysins were tested in the plate test with application of human, sheep, bovine and rabbit blood, incubating cultures at 22 and 37 degrees C for 3 days. Hemolysins of A. lwoffii were more active at 22 degrees C against rabbit and sheep erythrocytes. Direct hemolysins produced by remaining species of Acinetobacter hemolyzed most easily human erythrocytes. Out of 27 strains of A. haemolyticus, 26 produced hemolysins. Intermediate hemolysins, produced only by A. junii (43% of strains) and A. haemolyticus (37%), were active only against animal erythrocytes. A. baumanii and A. lwofii produced only intermediate hemolysins and percentage of strains producing them was, respectively, 3.6 and 7.0.

Acinetobacter↗

[Hemolytic properties of bacteria belonging to the Acinetobacter genus].

Direct and intermediate hemolytic activity of 526 strains of Acinetobacter was investigated. Their ability to produce lipase and lecithinase was also studied. Measurements were performed parallely on human, horse, sheep and bovine erythrocytes. Direct hemolytic activity was exhibited by 16% of tested strains (17 out of 24 strains of A. haemolyticus). Human, sheep and bovine erythrocytes were useful for testing the hemolytic activity of Acinetobacter. The hemolysis was occurring faster and was visible more frequently during incubation at 37 degrees C. Indirect hemolytic activity was observed in 88% of strains. Over 90% of strains were lipolytic after 24-48 hours which was independent of incubation temperature (22 and 37 degrees C).

Acinetobacter↗

[Sensitivity of Acinetobacter baumannii to antibiotics and disinfectants used in a Tunisian hospital milieu].

Seventy-nine strains of Acinetobacter baumannii isolated from pathologic products obtained mainly from surgical and intensive care units at the Rabta Hospital in Tunisia were studied. The sensitivity of these bacteria to the antibiotics used singly or in combination was tested in order to determine the most effective agents. Sensitivity to the disinfectants routinely used in our hospital was also studied using the "dilution-neutralisation" method according to "AFNOR" norms in order to identify the product with the most powerful preventive effect for Acinetobacter baumannii infection. The bacteria tested were resistant to all antibiotics except imipernem, colistin, and some aminoglycosides. A combination of imipenem-amikacin was effective against all strains. Best disinfection was obtained after 5 minutes of bacterial contact with aldehyde, phenol and amphotere.

Acinetobacter Infections↗

[Variation in transfer resistance to cefotaxime, ceftazidime and aztreonam in strains of Acinetobacter sp., Enterobacter sp. and Citrobacter sp. isolated from the same university hospitals].

BACKGROUND: In the last decade were in SR documented new problems in resistance to the newer antibiotics that with regard to their structure and antibacterial properties resisted to the known mechanism of bacterial resistance. The emergence of multiple drug resistance to the new betalactams is connected both with frequent application of these drugs in the hospitals and transfer of R plasmids. METHODS AND RESULTS: We studied composition and transferability of resistance to newer betalactam antibiotics in strains of Acinetobacter sp., Enterobacter sp, and Citrobacter sp. isolated during one month in patients from two teaching University Hospitals. All strains studied were resistant to cefotaxime (CTX), ceftazidime (CAZ) and aztreonam (ATM) but Acinetobacter strains, although resistant also to CAZ and ATM transferred the resistance to CTX only. Thus, resistance to CAZ and ATM has a chromosomal origin in these strains. A strain of Citrobacter sp., resistant to CTX, CAZ and ATM, produced a ESBL betalactamase detectable in a double-disk diffusion method (Fig. 1). An Enterobacter cloacae strain transfers directly the resistance to all new betalactams indicated. Their hydrolysis by these strains points to their production of new types of extended-spectrum beta-lactamases ESBL). CONCLUSIONS: We strongly recommend to be strictly rational in the use of new betalactams of CTX, CZA or ATM type because it is suspected that, especially in so-called empirical prophylaxis or treatment, they exert a strong selective pressure toward the prevalence of mutants with transferable ESBL-producing nosocomial bacteria resistant to these drugs.

Acinetobacter↗

[Kinetics of bactericidal activity of cefepime and cefpirome alone or combined with gentamicin, amikacin or ciprofloxacin against Acinetobacter baumannii, Stenotrophomonas maltophilia and Enterobacter cloacae hyperproductive in cephalosporinase].

Nosocomial infections encountered in intensive care units are frequently due to Gram negative bacilli among which Stenotrophomonas maltophilia, Acinetobacter sp., and Enterobacter sp. The aim of the present study was to evaluate the in vitro bactericidal activity of the new broad spectrum cephalosporins cefepime (FEP) and cefpirome (CPO) alone or in combination with amikacin (AKN), gentamicin (GTN) or ciprofloxacin (CIP) against Acinetobacter baumannii, Stenotrophomonas maltophilia and Enterobacter cloacae producing a derepressed cephalosporinase. This study was performed by using the time-kill curve method on 24 h with a starting inoculum of 10(6) - 10(7) cfu/ml. The combination of FEP (4 mg/l) with AKN (4 mg/l) against A. baumanii only results in about 1 log decrease at 24 h, but when FEP is combined at 8 mg/l, the decrease reaches 4 log in 24 h. The combination of FEP (16 and 32 mg/l) clavulanic acid (4 mg/l) resulted in 3 log decrease at 24 h. When combined with CIP 2 mg/l, FEP (16 and 32 mg) resulted in 5 and 6 log decrease in 24 h respectively. There were no survival bacteria at 6 h when FEP (32 mg/l) was combined with clavulanic acid (4 mg/l) and GTN (8 mg/l) at 6 h. Used alone FEP (1 mg/l) or CPO (1 mg/l) against E. cloacae, a 3 log decrease occurs at 6 h followed by a regrowth at 24 h. Combined with AKN (2 mg/l), FEP (1 mg/l) results in a 6 log decrease at 24 h, when CPO at 2 mg/l is needed for an equivalent result. These data show synergistic bactericidal activity of both new extended cephalosporins combined with AKN, GTN or CIP at concentrations achievable in biological fluid with adaptative dosage regimen.

Acinetobacter↗

[Cross-reactivity between Chlamydia psittaci and Acinetobacter in indirect immunofluorescence assays].

Cross reactivity between Chlamydia psittaci and a strain from genus Acinetobacter was investigated by indirect fluorescent antibody (IFA) technique. Two groups of serum samples were tested: 64 belonged to patients diagnosed as psittacosis and 64 (control group) were non reactive to Chlamydia psittaci by IFA. Samples were incubated on smears prepared with an Acinetobacter suspension for detecting IgG and IgM. 100% reactivity to IgG was found in 1:16 serum dilution among anti-psittacosis sera, whereas 6.25% of control sera reacted at the same dilution. When testing IgM, high rates of reactivity were found in both serum groups, thus it has been discarded as a marker for cross reactivity. Fluorescence pattern suggests that antigens are located at the cell wall.

Acinetobacter↗

Oleyl Oleate and Homologous Wax Esters Synthesized Coordinately from Oleic Acid by Acinetobacter and Coryneform Strains

Newly isolated Acinetobacter (NRRL B-14920, B-14921, B-14923) and coryneform (NRRL B-14922) strains accumulated oleyl oleate and homologous liquid wax esters (C30:2-C36:2) in culture broths. Diunsaturated oleyl oleate preponderated in 75 mg liquid wax esters (280 mg lipid extract) recovered from 100-ml cultures of Acinetobacter B-14920 supplemented with 810 mg oleic acid-oleyl alcohol. With soybean oil instead of oleic acid, wax esters (260 mg) were increased to approximately 50% of the lipid extract. Production of wax esters by cultures supplemented with combined fatty (C8-C18) alcohols and acids suggests a coordinated synthesis whereby the exogenous alcohol remains unaltered, and the fatty acid is partially oxidized with removal of C2 units before esterification. Consequently, C8-C18 primary alcohols control chain lengths of the wax esters. Exogenous fatty acids are presumed to enter an intracellular oxidation pool from which is produced a homologous series of liquid wax esters.

Journal Article↗

UV light-induced survival response in a highly radiation-resistant isolate of the Moraxella-Acinetobacter group.

A highly radiation-resistant member of the Moraxella-Acinetobacter group, isolate 4, obtained from meat, was studied to determine the effect of preexposure to UV radiation on subsequent UV light resistance. Cultures that were preexposed to UV light and incubated for a short time in plate count both exhibited increased survival of a UV light challenge dose. This response was inhibited in the presence of chloramphenicol. Frequencies of mutation to streptomycin, trimethoprim, and sulfanilamide resistance remained the same after the induction of this survival response and were not altered by treatment with mutagens, with the exception of mutation to streptomycin resistance after gamma-irradiation or nitrosoguanidine or methyl methane sulfonate treatment. The results indicated that isolate 4 has a UV light-inducible UV light resistance mechanism which is not associated with increased mutagenesis. The characteristics of the radiation resistance response in this organism are similar to those of certain other common food contaminants. Therefore, considered as part of the total microflora of meat, isolate 4 and the other radiation-resistant Moraxella-Acinetobacter isolates should not pose unique problems in a proposed radappertization process.

Anti-Bacterial Agents↗

Adhesion of acinetobacter venetianus to diesel fuel droplets studied with In situ electrochemical and molecular probes

The adhesion of a recently described species, Acinetobacter venetianus VE-C3 (F. Di Cello, M. Pepi, F. Baldi, and R. Fani, Res. Microbiol. 148:237-249, 1997), to diesel fuel (a mixture of C12 to C28 n-alkanes) and n-hexadecane was studied and compared to that of Acinetobacter sp. strain RAG-1, which is known to excrete the emulsifying lipopolysaccharide, emulsan. Oxygen consumption rates, biomass, cell hydrophobicity, electrophoretic mobility, and zeta potential were measured for the two strains. The dropping-mercury electrode (DME) was used as an in situ adhesion sensor. In seawater, RAG-1 was hydrophobic, with an electrophoretic mobility (&mgr;) of -0.38 x 10(-8) m2 V-1 s-1 and zeta potential (zeta) of -4.9 mV, while VE-C3 was hydrophilic, with &mgr; of -0.81 x 10(-8) m2 V-1 s-1 and zeta of -10.5 mV. The microbial adhesion to hydrocarbon (MATH) test showed that RAG-1 was always hydrophobic whereas the hydrophilic VE-C3 strain became hydrophobic only after exposure to n-alkanes. Adhesion of VE-C3 cells to diesel fuel was partly due to the production of capsular polysaccharides (CPS), which were stained with the lectin concanavalin A (ConA) conjugated to fluorescein isothiocyanate and observed in situ by confocal microscopy. The emulsan from RAG-1, which was negative to ConA, was stained with Nile Red fluorochrome instead. Confocal microscope observations at different times showed that VE-C3 underwent two types of adhesion: (i) cell-to-cell interactions, preceding the cell adhesion to the n-alkane, and (ii) incorporation of nanodroplets of n-alkane into the hydrophilic CPS to form a more hydrophobic polysaccharide-n-alkane matrix surrounding the cell wall. The incorporation of n-alkanes as nanodroplets into the CPS of VE-C3 cells might ensure the partitioning of the bulk apolar phase between the aqueous medium and the outer cell membrane and thus sustain a continuous growth rate over a prolonged period.

Journal Article↗

Enzymes of the tryptophan pathway in Acinetobacter calco-aceticus.

All enzymes of the tryptophan synthetic pathway were detectable in extracts from wild-type Acinetobacter calco-aceticus. The levels of these enzymes were determined in extracts from a number of auxotrophs grown under limiting tryptophan. In each case only anthranilate synthetase was found to be present in increased amounts, whereas the specific activities of the remaining enzymes remained unchanged and unaffected by the tryptophan concentration. Derepression of anthranilate synthetase was found to occur as the concentration of tryptophan became limiting. Anthranilate synthetase and phosphoribosyl transferase activities are both feedback-inhibited by tryptophan. Molecular weight determination carried out by gel filtration and zonal centrifugation in sucrose revealed that all the enzymes are less than 100,000, and no molecular aggregates of these enzymes were detected. The data indicate that tryptophan synthesis in Acinetobacter is regulated both by feedback inhibition of the first two enzymes of the pathway and by repression control of anthranilate synthetase.

Alkaline Phosphatase↗

Enzymes of the isoleucine-valine pathway in Acinetobacter.

Regulation of four of the enzymes required for isoleucine and valine biosynthesis in Acinetobacter was studied. A three- to fourfold derepression of acetohydroxyacid synthetase was routinely observed in two different wild-type strains when grown in minimal medium relative to cells grown in minimal medium supplemented with leucine, valine, and isoleucine. A similar degree of synthetase derepression was observed in appropriately grown isoleucine or leucine auxotrophs. No significant derepression of threonine deaminase or transaminase B occurred in either wild-type or mutant cells grown under a variety of conditions. Three amino acid analogues were tested with wild-type cells; except for a two- to threefold derepression of dihydroxyacid dehydrase when high concentrations of aminobutyric acid were added to the medium, essentially the same results were obtained. Experiments showed that threonine deaminase is subject to feedback inhibition by isoleucine and that valine reverses this inhibition. Cooperative effects in threonine deaminase were demonstrated with crude extracts. The data indicate that the synthesis of isoleucine and valine in Acinetobacter is regulated by repression control of acetohydroxyacid synthetase and feedback inhibition of threonine deaminase and acetohydroxyacid synthetase.

Alcaligenes↗

Isolation of outer membranes with an ordered array of surface subunits from Acinetobacter.

A method has been developed for the isolation of outer membranes from Acinetobacter sp. strain MJT/F5/199A. Washed cells were broken in a French press and, after deoxyribonuclease and ribonuclease treatment, removal of intact cells, and four washes in 20 mosmol phosphate buffer, pH 7.4, with centrifugation at 25,000 x g for 10 min, preparations of cell wall fragments from which almost all pieces of plasma membrane had been removed resulted. Treatment of the cell walls with lysozyme and further washing, in the presence of 20 mM MgCl(2), yielded preparations of outer membranes. Electron microscopy of freeze-etched preparations shows that a regular pattern of subunits is present on the outer surfaces of intact cells. After negative staining, these subunits are visible on isolated walls and outer membranes; they can be removed by brief treatment with papain. In section, the cell wall structure is that typical of gram-negative bacteria, but the subunits are not detectable on the surface of the outer membrane. The outer membrane retains the appearance of a "unit membrane" in the cell wall, isolated outer membrane, and papain-treated outer membrane fractions. Both cell walls and outer membranes contain a high percentage of protein (76 and 84%, respectively) and not more than 5% carbohydrate, of which glucose and galactose are constitutents. The outer membranes of this Acinetobacter thus differ in structure and composition from those of bacteria in the Enterobacteriaceae.

Alcaligenes↗

Chemical analysis of the outer membrane and other layers of the cell envelope of Acinetobacter sp.

Chemical analysis of fractions of the cell envelope of Acinetobacter sp. strain MJT/F5/199A, prepared by breakage in the French press and removal of plasma membranes, followed by sequential treatment with lysozyme and with papain, confirmed the existence of layers previously identified by electron microscopy. Outside the plasma membrane and periplasmic space, the envelope is composed of (i) a peptidoglycan-containing dense layer, (ii) an intermediate layer, (iii) a lipopolysaccharide-containing outer membrane, and (iv) an ordered array of protein subunits. A small amount of carbohydrate (3%) is found associated with protein in the fraction containing both the surface subunits and the intermediate layer. The papain-treated outer membranes contain 67% protein, 24% lipid, together with 11% lipopolysaccharide, and about 6% of non-lipopolysaccharide hexosamine. Lipid is located only in the papain-treated outer-membrane and is mainly phospholipid: 29% phosphatidyl glycerol, 30% phosphatidyl ethanolamine, and 40% cardiolipin. The principal fatty acid is C(18:1). Significant amounts of alcohols(16:1) and alcohols(18:1), which are found in Acinetobacter waxes, were recovered from the outer membrane.

Alcaligenes↗

Location of the fracture faces within the cell envelope of Acinetobacter species strain MJT-F5-5.

The cell wall of the gram-negative bacterium Acinetobacter species strain MJT/F5/5 shows in thin section an external "additional" layer, an outer membrane, an intermediate layer, and a dense layer. Negatively stained preparations showed that the additional layer is composed of hexagonally arranged subunits. In glycerol-treated preparations, freeze-etching revealed that the cell walls consist of four layers, with the main plane of fracture between layers cw 2 and cw 3. The surface of [Formula: see text] 2 consisted of densely packed particles, whereas [Formula: see text] 3 appeared to be fibrillar. In cell envelopes treated with lysozyme by various methods, the removal of the dense layer has detached the outer membrane and additional layer from the underlying layers, as shown in thin sections. When freeze-etched in the absence of glycerol, these detached outer membranes with additional layers fractured to reveal both the faces [Formula: see text] 2 and [Formula: see text] 3 with their characteristic surface structures, and, in addition, both the external and internal etched surfaces were revealed. This experiment provided conclusive evidence that the main fracture plane in the cell wall lies within the interior of the outer membrane. This and other evidence showed that the corresponding layers in thin sections and freeze-etched preparations are: the additional layer, cw 1; the outer membrane, cw (2 + 3); and the intermediate and dense layers together from cw 4. Because of similarities in structure between this Acinetobacter and other gram-negative bacteria, it seemed probable that the interior of the outer membrane is the plane most liable to fracture in the cell walls of most gram-negative bacteria.

Alcaligenes↗

Transformation of Acinetobacter calco-aceticus (Bacterium anitratum).

A highly efficient transformation system has been demonstrated in a strain of Acinetobacter calco-aceticus (Bacterium anitratrum). During mixed growth of various stable, unencapsulated, mutant strains, deoxyribonucleic acid (DNA) is liberated and fully encapuslated transformants can be isolated. Purified DNA preparations have been used to transform suitable recipient mutant strains for ability to synthesize capsules, ability to dispense with a growth factor requirement, and resistance to streptomycin. When the wild-type strain is deprived of its capsule, either by mechanical stripping or by mutation, the unencapsulated cells tend to form large clumped masses. A nonclumping mutant of an unencapsulated strain has been isolated. When ability to synthesize capsules is transformed into this nonclumping strain, the resultant cells no longer form chains, unlike the wild-type encapsulated strain. It appears likely that the occurrence of transformation during growth of mixed cultures, with glucose or gluconate as the carbon source, may be the result of osmotic rupture resulting from the inability of unencapsulated strains to oxidize triose phosphates as fast as they are formed. The finding of transformation in Acinetobacter may provide an additional useful organism for the study of this mode of genetic transfer since this strain grows well in a simple mineral medium containing a single oxidizable source of carbon. Furthermore, no special supplementary factors seem to be required for transformation to take place.

Alcaligenes↗

Two acyl-CoA dehydrogenases of Acinetobacter sp. strain M-1 that uses very long-chain n-alkanes.

Two genes encoding acyl-CoA dehydrogenases, acdA and acdB, arranged in tandem, were found in the chromosomal DNA of Acinetobacter sp. strain M-1. AcdA was purified from the parental strain and AcdB was purified from an Escherichia coli strain expressing the cloned gene. The substrate specificities of the two enzymes suggest that AcdA is a medium-chain acyl-CoA dehydrogenase and that AcdB is a long-chain acyl-CoA dehydrogenase. Characterization of n-alkane metabolism in Acinetobacter sp. strain M-1 has revealed parallel pathways as well as enzymes with overlapping specificities in a single pathway. The two acyl-CoA dehydrogenases described here provide another example of the physiological complexity underlying n-alkane utilization.

Journal Article↗

Treatment of multidrug resistant Acinetobacter.

PURPOSE OF REVIEW: Acinetobacter baumannii-calcoaceticus complex has become a serious nosocomial pathogen due to its persistence in the hospital environment and its broad antimicrobial resistance patterns. This review summarizes the most recent literature pertaining to the clinical management of infections with this bacteria emphasizing in-vitro antimicrobial resistance patterns and antimicrobial efficacy in animals and humans. RECENT FINDINGS: Although this pathogen can be associated with an elevated crude mortality, it only contributes to this mortality in a subset of high-risk patients. Determining in-vitro activity of antimicrobial agents can be problematic due to conflicting results sometimes obtained through different testing methods. There is no simple answer as to the most appropriate antimicrobial therapy secondary to lack of adequate studies. Imipenem/cilastatin, amikacin, ampicillin/sulbactam, colistin, rifampin, and tetracyclines are typically active against these bacteria. It is also not clear if combination therapy is more effective than monotherapy. In cases in which A. baumannii-calcoaceticus complex bacteria are resistant to all available agents, we have prolonged infusion times, increased drug dose, and altered route of instillation, such as nebulized therapy for pulmonary infections with mixed results. A primary goal of A. baumannii-calcoaceticus complex management should be to prevent initial colonization and subsequent infection by adequate infection control. SUMMARY: The A. baumannii-calcoaceticus complex continues to play a significant role in our healthcare systems. Prompt and adequate therapy with agents having in-vitro activity is required once it is established that the bacteria represents infection and not colonization. Aggressive infection control policies should be enforced when this pathogen is identified.

Acinetobacter Infections↗