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Natural transformation in Acinetobacter calcoaceticus.

Acinetobacter calcoaceticus is a metabolically versatile microorganism that is naturally competent for DNA uptake and incorporation. We have exploited the natural state of competency for studies involving the cloning, organization and expression of genes encoding catabolic enzymes. A. calcoaceticus is able to take up, at high efficiency, genetically engineered DNA, incorporate the DNA and stably maintain and express the DNA. Sequence analysis of cloned A. calcoaceticus DNA reveals a great deal of internal repetition and secondary structure, but no specific sequences associated with uptake appear to be present. Uptake and transformation occurs in solid and liquid medium, at a wide range of DNA concentrations and with little restriction barrier to the source of the transforming DNA.

Acinetobacter

[Isolation and characterization of rubredoxin from Acinetobacter calcoaceticus].

Acinetobacter calcoaceticus growing on long-chain n-alkanes contains a soluble iron-sulfur protein, which corresponds in its properties to a rubredoxin. It was prepared from the 50000 X g supernatant of ultrasonically treated cells using ion exchange chromatography on DEAE cellulose and gel filtration on Sephadex G-75. The isolated protein is pure electrophoretically, but yields two bands corresponding to molecular weights of 6000 and 12000 respectively. A content of 11 acidic against 6 basic amino acids is in line with the acidic character of the protein. The absence of acid-labile sulfur, content of 4 cysteine residues and one iron atom per polypeptide chain and the typical absorption maxima at gamma = 280, 380, and 490 nm exclude the presence of a ferredoxin. Involvement of the rubredoxin in the alkane hydroxylation is discussed.

Acinetobacter

Regulation of enzyme synthesis in the tryptophan pathway of Acinetobacter calcoaceticus.

In Acinetobacter calcoaceticus the seven genes coding for the enzymes responsible for tryptophan synthesis map at three chromosomal locations. Two three-gene clusters, one (trpGDC) specifying the small subunit of anthranilate synthase, phosphoribosyl transferase, and indoleglycerol phosphate synthase and the other (trpFBA) specifying phosphoribosyl anthranilate isomerase and both tryptophan synthase subunits, are not linked to each other or to the trpE gene specifying the large anthranilate synthase subunit. When regulation of trp gene expression is studied in the wild type, only the level of the trpF gene product decreases upon addition of tryptophan to the medium. Tryptophan starvation of tryptophan auxotrophs, however, results in increased levels of all the tryptophan enzymes; this and additional evidence suggests that the expression of all the trp genes is subject to repression. The trpGDC genes are coordinately controlled, and the trpE gene is regulated in parallel with them. The trpFBA genes are controlled neither coordinately nor in parallel with the other trp genes, but respond proportionally when compared with each other. So far, two types of constitutive mutants have been found. The first class of mutants apparently occurs in the structural gene for a repressor protein; this repressor locus is unlinked to any of the biosynthetic trp genes and affects only the expression of trpE and the trpGDC cluster. The second class contains mutants closely linked to the trpGDC region; they overproduce only the gene products of this cluster.

Acinetobacter

[A case of meningitis due to Acinetobacter calcoaceticus].

Meningitis due to Acinetobacter calcoaceticus is rare. These rare bacteria were isolated from culture fluid in patient resembling to Neisseria meningitis with clinical and laboratory findings. The patient was treated with sensitive antibiotic.

Acinetobacter Infections

Equine myositis and septicemia caused by Acinetobacter calcoaceticus infection.

Myositis and septicemia caused by Acinetobacter calcoaceticus were diagnosed in a mare. The infection was characterized clinically by ventral swelling and edema, diarrhea, listlessness, and rectal temperature of 39.4 C. The mare was treated symptomatically for 2 days but died on the 3rd day. Conditions seen at necropsy were myositis, enteritis, typhlitis, colitis, and hepatitis. Lymph nodes were moderately enlarged throughout the body. Gross lesions in musculature were edema, scarring, petechiae, and an occasional exxhymosis. The enteritis was catarrhal, with excessive mucus and moderate hyperemia. The typhlitis and colitis were hemorrhagic. The swollen liver had a diffuse mottled pale and red pattern. Microscopic lesions in skeletal muscle consisted of petechiation, necrosis, scarring, and edema. Cardiac muscle was also scarred and necrotic, but edema was not prominent. Periacinal necrosis was found in the liver. Acinetobacter calcoaceticus was isolated from myocardium and liver.

Acinetobacter Infections

[Cytochrome composition of Acinetobacter calcoaceticus].

The qualitative and quantitative composition of cytochromes in intact cells of Acinetobacter calcoaceticus and in particle fractions obtained from cells following ultrasonic treatment by differential centrifugation were studied using spectrophotometric methods. Acinetobacter calcoaceticus contains cytochrome b, cytochrome o and low amounts of cytochrome d. Both the absolute content of cytochrome b and o and the relative composition do not essentially vary with the carbon source used (hexadecane, acetate, succinate, malate, yeast extract). Only bacterial cultivated on yeast extract show, under simultaneous decrease of the content of cytochrome o, an increased formation of cytochrome d. In Acinetobacter, cytochromes appear not to be immediately involved in n-alkane hydroxylation.

Acinetobacter

In vitro synergism between carbenicillin and aminoglycosidic aminocyclitols against Acinetobacter calcoaceticus var. anitratus.

Acinetobacter caleoaceticus var. anitratus is a nonfermentative, gram-negative bacillus that has been demonstrated to cause severe infections, usually in hospitalized patients. Since mild to moderate resistance of A. calcoaceticus to one or more aminoglycosidic aminocyclitols has been noted to occur, a study was undertaken to evaluate the activity of combinations of carbenicillin with either kanamycin, tobramycin, or gentamicin against 28 isolates of A. calcoaceticus obtained from clinical sources. Synergism (defined as at least 100-fold-increased killing at 24 h by the combination as compared with the most efficacious of the individual antibiotics) was demonstrated against 26 of 28 strains of A. calcoaceticus with carbenicillin plus kanamycin and carbenicillin plus tobramycin and against 25 of 28 strains with carbenicillin plus gentamicin. The median increased killing for the 28 strains was 4.2 log(10) with carbenicillin plus kanamycin and with carbenicillin plus tobramycin and 3.1 log(10) with carbenicillin plus gentamicin. The most important determinant of synergistic potential of each combination was the level of resistance of each strain of A. calcoaceticus to the aminoglycoside component of the combination.

Acinetobacter

Ventriculitis from Acinetobacter calcoaceticus variant anitratus.

Ventriculitis from infection with Acinetobacter calcoaceticus variant anitratus occurred in an infant two weeks after surgical repair of a lumbosacral meningomyelocele and ventriculoperitoneal shunting. Recovery took place on replacing the shunt with an extraventricular deviation device and giving gentamicin and ampicillin intravenously.

Acinetobacter Infections

Acinetobacter calcoaceticus variety anitratus: an increasing nosocomial problem.

Acinetobacter calcoaceticus var anitratus is an increasing nosocomial problem in some institutions. Duirng a recent 16-month period, we observed 15 patients with blood isolates of this organism. Thirteen of these patients acquired the organism from the hospital environment. Eleven of these represented real disease and concurrent growth of the organism from another site was predictive of infection (P less than 0.03). A review of laboratory records showed that there was a three-fold increase in nonblood isolates of this organism from hospitalized patients in 1975 and 1976, as compared to 1972 (P less than 0.0001), and a marked seasonal effect was noted, with increased isolations during the summer. Aminoglycoside resistance had increased considerably with 25% of nonblood isolates being resistant to gentamicin and 16% resistant to tobramycin despite its restriction; blood isolates were uniformly susceptible to gentamicin and tobramycin. Reporting of A anitratus as a nosocomial pathogen and serotyping of isolates may be useful to further define its role in nosocomial infections.

Acinetobacter Infections

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

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

Acinetobacter

Serological cross-reactions between Acinetobacter calcoaceticus and chlamydiae.

A cross-reaction between Acinetobacter calcoaceticus and chlamydiae is described. A water-soluble, heat stable, non-dialyzable antigen was extracted from Acinetobacter species by boiling. This antigen fixed complement in the presence of homologous hyperimmune sera from rabbits or guinea pigs and in the presence of heterologous human or hyperimmunized animal sera containing chlamydial antibodies. Hyperimmune antisera to the extracted antigen, or to suspensions of live acinetobacters, also reacted in complement fixation with a group-specific antigen.

Acinetobacter

Assimilatory nitrate reductase from Acinetobacter calcoaceticus.

A soluble nitrate reductase from the bacterium Acinetobacter calcoaceticus grown on nitrate has been characterized. The reduction of nitrate to nitrite is mediated by an enzyme of 96000 molecular weight that can use as electron donors either viologen dyes chemically reduced with dithionite or enzymatically reduced with NAD(P)H, through specific diaphorases which utilize viologens as electron acceptors. Nitrate reductase activity is molybdenumdependent as shown by tungstate antagonistic experiments and is sensitive to--SH reagents and metal chelators such as KCN. The enzyme synthesis is repressed by ammonia. Moreover, nitrate reductase activity undergoes a quick inactivation either by dithionite and temperature or by dithionite in the presence of small amounts of nitrate. Cyanate prevents this inactivating process and can restore the activity once the inactivation had occurred, thus suggesting that an interconversion mechanism may participate in the regulation of Acinetobacter nitrate reductase.

Acinetobacter

[Purification and various properties of NADP+-dependent alcohol dehydrogenase from Acinetobacter calcoaceticus].

The constitutive NADP+-dependent alcohol dehydrogenase from Acinetobacter calcoaceticus can be accumulated about 50 fold in 3 purification steps. The end-product shows in the analytical polyacrylamide gel electrophoresis only one active enzyme band. The molecular weight of the enzyme was determined to be 235,000 by gel chromatography on Sephadex G 200, the smallest subunit shows a molecular weight of 61 000 on SDS electrophoresis. The isoelectric point is at 5.84. The KM values determined with primary aliphatic alcohols diminish in the range of the homologous order (C2--C10) with growing chain length. The KM value for hexanal is about 20 fold less than that for 1-hexanol.

Acinetobacter

Correlation between twitching motility and possession of polar fimbriae in Acinetobacter calcoaceticus.

Negatively stained preparations of 16 strains of Acinetobacter calcoaceticus were examined in the electron microscope. Five of the strains did and 11 of the strains did not exhibit twitching motility. Two of the non-twitching strains were substrains of twitching wild-type strains. All twitching strains were found to possess fimbriae with a diameter of approximately 50 A. These fimbriae were shown to be of polar origin except in one strain where, for technical reasons, the origin could not be determined with certainty. Polar fimbriae could not be demonstrated in any of the strains that did not exhibit twitching motility. The demonstration of polar fimbriae was only regularly possible during the exponential growth phase, presumably because the fimbriae are shed by the bacteria during later growth phases. During the study, methods were developed for securing exponentially growing bacteria in concentrations suited for the preparation of negatively stained cells for electron microscopy. Peritrichously arranged fimbriae with a diameter of approximately 30 A were demonstrated in nearly all strains studied, especially on cells in late growth phases. The occurrence of this kind of fimbriae was therefore not correlated with the occurrence of twitching motility.

Acinetobacter

[Utilization of trimethylammonium-compounds by Acinetobacter calcoaceticus (author's transl)].

The utilization of carnitine and carnitine derivatives (O-acylcarnitines, carnitine carboxylderivatives) and structure-related trimethylammonium-compounds (betaines and nitrogen-bases) by Acinetobacter calcoaceticus was studied by means of the control of growth and the quantitative detection of metabolites. The strain grew only on L-carnitine, L-O-acylcarnitines, and gamma-butyrobetaine as the sole carbon sources. The utilization of these compounds and the growth correlated with the cleavage of the C-N bond and thereby with the formation of trimethylamin. D-Carnitine was metabolized, if an additional carbon source, like L-carnitine, was present in the incubation mixture, or if the bacteria were preincubated with L- or DL-carnitine, but no growth was observed on D-carnitine as the sole carbon source. The bacteria oxidized choline to glycinebetaine in the presence of additional carbon sources, glycinebetaine itself was not assimilated. With regard to the catabolism of quaternary nitrogen compounds Acinetobacter calcoaceticus shows a different pathway in comparison with other bacterial species metabolizing carnitine.

Acinetobacter

[Inhibiton of isocitrate dehydrogenase and isocitrate lyase from Acinetobacter calcoaceticus by acids of the citrate and glyoxylate cycle].

Acinetobacter calcoaceticus contains two forms of NADP+-dependent isocitrate dehydrogenases differing, among others, by their molecular weights and regulatory properties. The regulation of the high-molecular form of isocitrate dehydrogenase and of isocitrate lyase by organic acids, either belonging or related to the citrate and glyoxalate cycle, is investigated. While alpha-ketoglutarate and oxalacetate competitively inhibit the isocitrate dehydrogenase against Ds-isocitrate, glyoxylate and pyruvate were found to increase Vmax and to lower the KM value for Ds-isocitrate and NADP+. Simultaneous addition of oxalacetate and glyoxylate (not, however, addition of the nonenzymatically formed condensation product of both compound) nullified the activation of isocitrate dehydrogenase by glyoxylate, and potentiates the inhibitory effect of oxalacetate. Alpha-ketoglutarate, succinate, and phosphoenolpyruvate inhibit the isocitrate lyase in a noncompetitive fashion against DS-isocitrate; L-malate, oxalacetate and glyoxylate inhibit competitively. The intermediates of the citrate and glyoxylate cycle afford additive inhibition of the isocitrate lyase. The importance of organic acids of the citrate and glyoxylate cycle and of phosphoenolpyruvate for the regulation of the citrate and glyoxylate cycle at the level of isocitrate dehydrogenase and isocitrate lyase is discussed.

Acinetobacter

Community-acquired Acinetobacter calcoaceticus var anitratus pneumonia.

Two patients had community-acquired Acinetobacter calcoaceticus var anitratus pneumonia. Both patients were alcoholic and one was cirrhotic. One patient died and the other received two weeks of gentamicin therapy and survived. Misinterpretation of the sputum Gram stain delayed diagnosis and institution of proper therapy in both cases. In addition to organisms sensitive to penicillins such as Neisseria or Haemophilus, Acinetobacter must be considered in the differential diagnosis of community-acquired Gram-negative coccobacillary pneumonia.

Acinetobacter