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PubMed2002

Pseudomonas indica sp. nov., a novel butane-utilizing species.

The taxonomic position of two butane-utilizing bacteria was studied using a polyphasic approach. Biochemical and physiological characteristics indicated these to be members of the genus Pseudomonas, showing more similarity to Pseudomonas mendocina than to any other species. The major fatty acids found in these two strains also pointed to their similarity to P. mendocina. On the other hand, DNA-DNA hybridization studies with seven related Pseudomonas species belonging to the gamma-Proteobacteria and the deltaTm values of reassociated molecules clearly showed that these two strains do not belong to any of the seven species tested. The 16S rRNA gene was sequenced and compared with the sequences available in the GenBank database. Phylogenetic analysis using the region covering positions 31-1488 (Escherichia coli numbering) confirmed these observations and placed these two strains as members of the authentic Pseudomonas, but not in any existing species of the genus. On the basis of biochemical characteristics, fatty acid profiles, DNA-DNA reassociation and deltaTm values, as well as 16S rRNA gene sequence analyses, these two isolates were shown to belong to one species but to have characteristics distinct from those of validly described species of Pseudomonas (sensu stricto). These strains, therefore, should be recognized as a novel species, for which the name Pseudomonas indica sp. nov. is proposed. The type strain is strain IMT37T (= MTCC 3713T = DSM 14015T).

Base Sequence↗

PubMed1981

Interactions of Pseudomonas aeruginosa lectins with Escherichia coli strains bearing blood group determinants.

Pseudomonas aeruginosa lectins interact with Escherichia coli strains O86B7 and O128B12, which possess B and H (O) blood group determinants, respectively. The interaction could be demonstrated by specific agglutination of the bacteria, by haemagglutination inhibition tests and by lectin-mediated peroxidase binding to the bacteria. The agglutination of E. coli O86B7 by the Pseudomonas galactose-binding lectin was inhibited by D-galactose and by the lipopolysaccharide extracted from E. coli O86B7. Similarly, the specific agglutination of E. coli O128B12 by the Pseudomonas mannose-binding lectin (which also binds L-fucose, L-galactose and D-fructose) was inhibited by D-mannose, L-fucose, L-galactose and D-fructose, as well as by athe lipopolysaccharide extracted from E. coli O128B12. The interaction between E. coli O128B12 and the Pseudomonas mannose-binding lectin was also demonstrated by lectin-mediated peroxidase binding to the bacterial surface. Peroxidase binding was also inhibited by the above-mentioned sugars and E. coli O128B12 lipopolysaccharide. Treatment of cells of the two E. coli strains with protein-denaturing agents did not reduce their agglutination by the Pseudomonas lectins. On the other hand, oxidation of the cell surface sugars by sodium metaperiodate or boiling the cells in the presence of 1% acetic acid for 1 h abolished their agglutination by the two lectins. It is, therefore, suggested that the Pseudomonas lectins interact with the B and H (O) blood group determinant sugars (D-galactose in E. coli O86B7 and L-fucose in E. coli O128B12) residing in the lipopolysaccharides of these E. coli strains.

ABO Blood-Group System↗

PubMed1989

13C-NMR studies of acetate and methanol metabolism by methylotrophic Pseudomonas strains.

The metabolism of [2-13C]acetate by Pseudomonas M27(Icl-) and Pseudomonas MA(Icl+) was studied in vivo using 13C-NMR spectroscopy. The flux of 13C-label into bicarbonate, glutamate and citrate was observed in both organisms. In addition 13C-labelled alpha, alpha-trehalose was synthesized as a major metabolite by Pseudomonas M27 but not by Pseudomonas MA. The presence of this disaccharide in cell extracts of Pseudomonas AM1(Icl-) grown with [13C]methanol was also observed. The data from analysis of the trehalose multiplet signal observed in the spectra of Pseudomonas M27 cell extracts were consistent with the absence of the glyoxylate cycle in this methylotroph.

Acetates↗

PubMed2005

Pseudomonas azotifigens sp. nov., a novel nitrogen-fixing bacterium isolated from a compost pile.

A nitrogen-fixing bacterium, designated strain 6H33b(T), was isolated from a compost pile in Japan. The nitrogenase activity of this strain was detected based on its acetylene-reducing activity under low oxygen concentrations (2-4%). An analysis of the genes responsible for nitrogen fixation in this strain, nifH and nifD, indicated a close relationship to those of Pseudomonas stutzeri A15 (A1501). Sequence similarity searches based on the 16S rRNA gene sequences showed that strain 6H33b(T) belongs within the genus Pseudomonas sensu stricto; closest similarity was with Pseudomonas indica (97.3%). A comparison of several taxonomic characteristics of 6H33b(T) with those of P. indica and some type strains of the genus Pseudomonas sensu stricto indicated that 6H33b(T) could be distinguished from P. indica based on the presence of nitrogen fixation ability, the absence of nitrate reduction and denitrification abilities and the utilization of some sugars and organic acids. Phylogenetic analyses and the results of DNA-DNA hybridization experiments also indicated that strain 6H33b(T) represents a species distinct from P. indica. From these results, it is proposed that strain 6H33b(T) (=ATCC BAA-1049(T)=JCM 12708(T)) is classified as the type strain of a novel species of the genus Pseudomonas sensu stricto under the name Pseudomonas azotifigens sp. nov.

Bacterial Typing Techniques↗

PubMed2006

Pseudomonas segetis sp. nov., isolated from soil.

A Gram-negative, aerobic bacterium, designated strain FR1439T, was isolated from the soil of Dokdo in the Republic of Korea. The cells of strain FR1439T were catalase- and oxidase-positive, motile and rod-shaped. Phylogenetic analysis of the 16S rRNA gene sequence revealed that it represents a distinct line of descent within the genus Pseudomonas. The levels of DNA-DNA relatedness between strain FR1439T and type strains of phylogenetically related species, namely Pseudomonas flavescens, Pseudomonas mendocina, Pseudomonas pseudoalcaligenes and Pseudomonas straminea, ranged from 28 to 37%. Several phenotypic characteristics, together with the cellular fatty acid composition, can be used to differentiate strain FR1439T from related pseudomonads. On the basis of the polyphasic taxonomic evidence presented in this study, strain FR1439T represents a novel species, for which the name Pseudomonas segetis sp. nov. is proposed. The type strain is FR1439T (=IMSNU 14101T=CIP 108523T=KCTC 12331T).

Bacterial Typing Techniques↗

PubMed2006

Decreased abundance and diversity of culturable Pseudomonas spp. populations with increasing copper exposure in the sugar beet rhizosphere.

Recent studies have indicated that culturable bacteria constitute highly sensitive bioindicators of metal-induced stress in soil. We report the impact of different copper exposure levels characteristic of contaminated agricultural soils on culturable Pseudomonas spp. in the rhizosphere of sugar beet. We observed that the abundance of Pseudomonas spp. was much more severely affected than that of the general population of culturable heterotrophic bacteria by copper. For diversity assessment, Pseudomonas isolates were divided into operational taxonomic units based on amplified ribosomal DNA restriction analysis and genomic PCR fingerprinting by universally primed PCR. Copper significantly decreased the diversity of Pseudomonas spp. in the rhizosphere and significantly increased the frequency of copper-resistant isolates. Concomitant chemical and biological analysis of copper in the rhizosphere and in bulk soil extracts indicated no rhizosphere effect and a relatively low copper bioavailability in the studied soil, suggesting that the observed effects of copper may occur at lower total concentrations in other soils. We conclude that culturable Pseudomonas sensu stricto constitutes a highly sensitive and relevant bioindicator group for the impact of copper in the rhizosphere habitat, and suggest that continued application of copper to agricultural soils poses a significant risk to successful rhizosphere colonization by Pseudomonas spp.

Beta vulgaris↗

PubMed1990

Comparative in vitro activities of newer quinolones against Pseudomonas species and Xanthomonas maltophilia isolated from patients with cancer.

The in vitro susceptibilities of three Pseudomonas species (Pseudomonas aeruginosa, Pseudomonas putida, and Pseudomonas fluorescens) and Xanthomonas maltophilia to quinolone antimicrobial agents were determined. Several newer agents, particularly PD117558, PD117596, PD127391, sparfloxacin (AT-4140), A-56620, and temafloxacin, were active against Pseudomonas species. X. maltophilia isolates were generally less susceptible than were Pseudomonas isolates but were inhibited by some of the newer quinolones.

4-Quinolones↗

PubMed1989

Selective medium for Pseudomonas aeruginosa that uses 1,10-phenanthroline as the selective agent.

The MIC of 1,10-phenanthroline for 35 Pseudomonas aeruginosa strains was 128 micrograms/ml, whereas 32 micrograms or less per ml inhibited all other microorganisms tested. On the basis of these results, a selective agar for P. aeruginosa which contained 15 g of Trypticase soy broth (BBL Microbiology Systems), 15 g of agar, and 0.1 g of phenanthroline per liter was formulated. Forty-four P. aeruginosa strains yielded a mean efficiency of plating on this medium of 79% of the counts obtained on Trypticase soy agar, which was significantly higher than that obtained with pseudomonas isolation agar or Pseudosel agar. Pseudomonas cepacia, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas stutzeri, representatives of 13 other genera (including gram-negative rods, gram-positive rods, and cocci), and a yeast were not recovered within 48 h at 35 degrees C when approximately 10(7) CFU were plated on this medium. Only small colonies from one strain each of P. fluorescens and P. putida could be seen at 3 and 7 days, respectively, and they had an efficiency of plating of only less than 0.001%. When 10(7) CFU of either of these strains was plated with 10(2) CFU of P. aeruginosa, it did not interfere with the quantitative recovery of P. aeruginosa.

Culture Media↗

PubMed2002

Characterization of Pseudomonas spp. associated with spoilage of gilt-head sea bream stored under various conditions.

The population dynamics of pseudomonads in gilt-head sea bream Mediterranean fish (Sparus aurata) stored under different conditions were studied. Phenotypic analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins were performed to identify a total of 106 Pseudomonas strains isolated from S. aurata stored under different temperatures (at 0, 10, and 20 degrees C) and packaging conditions (air and a modified atmosphere of 40% CO(2)-30% N(2)-30% O(2)). Pseudomonas lundensis was the predominant species, followed by Pseudomonas fluorescens, while Pseudomonas fragi and Pseudomonas putida were detected less frequently. Fluorescent Pseudomonas strains dominated under air conditions, while proteolytic and less lipolytic strains dominated under modified-atmosphere packaging. Different storage conditions appear to govern the selection of pseudomonads in gilt-head sea bream fish.

Animals↗

PubMed1980

Resistance of exotoxin A to purified Pseudomonas proteolytic enzymes.

Pseudomonas toxin is produced as a proenzyme which is cytotoxic for cells in culture but must be activated to express full enzymatic activity. The ability of purified pseudomonas alkaline protease and elastase or of culture filtrates of two strains of Pseudomonas aeruginosa to modify the activity of pseudomonas toxin was examined. Two parameters of toxin activity were followed: enzymatic activity, i.e., the adenosine diphosphate (ADP) ribosylation of elongation factor 2, and biological activity, i.e., inhibition of protein synthesis in cultured mouse fibroblasts. Biological activity of toxin depends upon an intact toxin molecule, whereas enzyme activity requires only a functional A region. Incubation with purified pseudomonas proteolytic enzymes did not alter either enzymatic or biological activity. The toxin is not refractory to the action of all proteolytic enzymes, since thermolysin rapidly destroyed the toxin molecule. Treatment of toxin with culture filtrates of P. aeruginosa reduced ADP ribosylation activity, but increased the ability of toxin to inhibit protein synthesis in cell monolayers. Incubation of culture filtrates with one of the protease inhibitors alpha-2-macroglobulin or phosphoramidon did not alter the effect of the filtrates on biological activity. Alpha-2-macroglobulin, however, caused a fourfold stimulation of ADP ribosylation activity of the toxin. We conclude that pseudomonas alkaline protease and elastase are not responsible for the modifications in toxin activity induced by culture filtrates of P. aeruginosa; the factors responsible have not yet been identified, but are not inactivated by phosphoramidon or alpha-2-macroglobulin.

Adenosine Diphosphate↗

PubMed1980

Homologous structural genes and similar induction patterns in Azotobacter spp. and Pseudomonas spp.

Intergeneric comparison of the three enzymes that initiate metabolism of protocatechuate in Azotobacter and Pseudomonas species revealed close immunological relatedness of isofunctional proteins. Furthermore, beta-ketoadipate induces all of the enzymes of the protocatechuate pathway (except protocatechuate oxygenase) in Azotobacter and in Pseudomonas species of the "fluorescent" and "cepacia" groups. This regulatory property sets the organisms apart from other bacteria. Protocatechuate oxygenase from Pseudomonas cepacia, like the enzyme from fluorescent Pseudomonas species, cross-reacts strongly with antiserum prepared against protocatechuate oxygenase from Azotobacter vinelandii. Double-diffusion experiments conducted with the antiserum revealed relatedness of Azotobacter spp. Protocatechuate oxygenases in the following order: A. vinelandii = Azotobacter miscellum greater than Azotobacter chroococcum greater than Azotobacter beijerinkii. The antiserum also revealed serological heterogeneity among Pseudomonas spp. protocatechuate oxygenases which were serologically indistinguishable in earlier studies using Pseudomonas aeruginosa protocatechuate oxygenase as reference protein.

Azotobacter↗

PubMed2001

Regulation of rpoS gene expression in Pseudomonas: involvement of a TetR family regulator.

The rpoS gene encodes the sigma factor which was identified in several gram-negative bacteria as a central regulator during stationary phase. rpoS gene regulation is known to respond to cell density, showing higher expression in stationary phase. For Pseudomonas aeruginosa, it has been demonstrated that the cell-density-dependent regulation response known as quorum sensing interacts with this regulatory response. Using the rpoS promoter of P. putida, we identified a genomic Tn5 insertion mutant of P. putida which showed a 90% decrease in rpoS promoter activity, resulting in less RpoS being present in a cell at stationary phase. Molecular analysis revealed that this mutant carried a Tn5 insertion in a gene, designated psrA (Pseudomonas sigma regulator), which codes for a protein (PsrA) of 26.3 kDa. PsrA contains a helix-turn-helix motif typical of DNA binding proteins and belongs to the TetR family of bacterial regulators. The homolog of the psrA gene was identified in P. aeruginosa; the protein showed 90% identity to PsrA of P. putida. A psrA::Tn5 insertion mutant of P. aeruginosa was constructed. In both Pseudomonas species, psrA was genetically linked to the SOS lexA repressor gene. Similar to what was observed for P. putida, a psrA null mutant of P. aeruginosa also showed a 90% reduction in rpoS promoter activity; both mutants could be complemented for rpoS promoter activity when the psrA gene was provided in trans. psrA mutants of both Pseudomonas species lost the ability to induce rpoS expression at stationary phase, but they retained the ability to produce quorum-sensing autoinducer molecules. PsrA was demonstrated to negatively regulate psrA gene expression in Pseudomonas and in Escherichia coli as well as to be capable of activating the rpoS promoter in E. coli. Our data suggest that PsrA is an important regulatory protein of Pseudomonas spp. involved in the regulatory cascade controlling rpoS gene regulation in response to cell density.

Amino Acid Sequence↗

PubMed1987

Successful treatment of Pseudomonas peritonitis during continuous ambulatory peritoneal dialysis.

Successful eradication of Pseudomonas peritonitis is described in 12 (57%) of 21 cases from a large continuous ambulatory peritoneal dialysis (CAPD) program at a tertiary care center. In successful cases, cure was achieved within 17 days using therapy which included aminoglycoside started routinely at the onset of symptoms and an antipseudomonal penicillin or cephalosporin derivative added as soon as pseudomonas infection was identified on culture. Of the 9 treatment failures which required catheter removal, 2 had failure of peritoneal drainage, 4 had infection with multiple and/or drug-resistant Pseudomonas strains, and 3 had persistent catheter tunnel infection which resulted in recurrent Pseudomonas peritonitis. Factors such as diabetes mellitus and pediatric age group did not prevent successful medical therapy. Predisposing factors favoring development of Pseudomonas peritonitis included technical failures and in a few cases recent antibiotic therapy. We conclude that Pseudomonas peritonitis complicating CAPD can be successfully cured without catheter removal or discontinuation of CAPD in many cases, particularly when complicating factors are not present.

Adolescent↗

PubMed2005

Pseudomonas xanthomarina sp. nov., a novel bacterium isolated from marine ascidian.

Two Pseudomonas-like yellow-orange-pigmented non-fluorescent denitrifying strains KMM 235 and KMM 1447T were isolated from marine ascidian specimens and investigated by a polyphasic approach to clarify their taxonomic status. On the basis of 16S rDNA gene sequence data the new isolates clustered with the Pseudomonas stutzeri species group with sequence similarities of >98%. The results of DNA-DNA hybridization and biochemical characterization showed genetic and phenotypic distinction between strains KMM 235 and KMM 1447T and from the other validly described Pseudomonas species. Strain KMM 235 was found to be closely related to the type strain of Pseudomonas stutzeri in their phenotypic and genetic characteristics and represented, probably, a new P. stutzeri genomovar. It is proposed that strain KMM 1447T be classified as a new species of the genus Pseudomonas, Pseudomonas xanthomarina sp. nov., with the type strain KMM 1447T (=JCM 12468T=NRIC 0617T=CCUG 46543T).

Animals↗

PubMed2002

Effects of some plant extracts and antibiotics on Pseudomonas aeruginosa isolated from various burn cases.

OBJECTIVE: To determine the major groups of bacteria associated with burn infections, isolation of Pseudomonas aeruginosa from burn cases, and the testing of antibacterial activity of some plant extracts in comparison with the standard antibiotics. METHODS: A total of 92 burn cases, covered with tetracycline (40 cases) and non-covered with tetracycline (52 cases) from both sex and various ages, were collected from various hospitals in Basrah City, Iraq on the year 2001. Bacteriological investigation for isolation of bacterial pathogens especially Pseudomonas aeruginosa was carried out. Two methods were used to evaluate the anti-bacterial activity of various concentrations of aqueous extracts of leaves of Myrtus communis and Eucalyptus with comparison to 6 antibiotics, these methods were to determine the growth inhibition zones and minimal inhibitory concentration. RESULTS: From a total 92 cases, 78 (84.8%) gave positive cultures. One hundred and fifty eight isolates were identified as bacterial pathogens. Pseudomonas aeruginosa was the predominant bacterial pathogen from 35.9% of the tetracycline covered burns and 42.3% of the non-tetracycline covered burns followed by other bacterial types in various percentages. Aqueous leaves extracts of myrtus communis and eucalyptus gave an excellent effect on bacterial growth and their effects were located within the limits of antibiotic effects. CONCLUSION: Pseudomonas aeruginosa was the predominant bacterial isolate followed by Staphylococcus aureus, then other type of bacterial was isolated in various percentage from each tetracycline covered burns and non-tetracycline covered burns. Most concentrations of the extracts of the studied plants showed a high antibacterial activity against Pseudomonas aeruginosa, and showed significant differences between susceptibility of Pseudomonas aeruginosa isolated from each tetracycline covered burn and non-tetracycline covered burn.

Anti-Bacterial Agents↗

PubMed1992

[Effect of cefepime on adhesion and phagocytosis of Pseudomonas aeruginosa].

BACKGROUND: We studied the effect of subinhibitory concentrations of cefepime on cell surface hydrophobicity expression and adherence to epithelial cells of Pseudomonas aeruginosa. We also evaluated the effect of this drug on the interaction between Pseudomonas aeruginosa and human polymorphonuclear leukocytes. METHODS: The bacterial cell surface hydrophobicity was tested on a biphasic system (water/p-xylene). The bacterial adherence to epithelial cells as well as the phagocytosis of Pseudomonas aeruginosa was studied using radiometric techniques. The superoxide production was evaluated by means of cytochrome C reduction method. RESULTS: Pre-incubation of a non mucoid Pseudomonas aeruginosa strain with a 0.25 MIC value of cefepime produces a reduction in the adherence of this microorganism to epithelial cells, but without modifying the surface hydrophobicity. Pre-incubation of a mucoid and a non mucoid P. aeruginosa strain with a 0.25 MIC value of cefepime reduces its phagocytosis by polymorphonuclear leukocytes. The preincubation polymorphonuclear leukocytes with cefepime (1, 10, and 100 mg/ml) did not influence the phagocytosis of Pseudomonas aeruginosa nor the bactericidal mechanisms of PMN (superoxide production). At therapeutic concentrations, cefepime showed intracellular activity against P. aeruginosa in human polymorphonuclear leukocytes. CONCLUSIONS: The incubation of Pseudomonas aeruginosa with sub-MIC inhibit its adherence to epithelial cells and its intake by human polymorphonuclear leukocytes. At therapeutic concentrations, cefepime did not affect the bactericidal mechanisms of polymorphonuclear leukocytes, and shows a significant intracellular activity against P. aeruginosa.

Bacterial Adhesion↗

PubMed2003

Mycotic ascending aortic pseudoaneurysm secondary to pseudomonas mediastinitis at the aortic cannulation site.

During the last 5 years, postoperative Pseudomonas mediastinitis has occurred in 2 of the 3,072 patients in our institution who have undergone cardiopulmonary bypass cardiac operations via a sternotomy. To our knowledge, there is no prior report in the English-language literature of postoperative Pseudomonas mediastinitis that originated at the aortic cannulation site, yet that was the site of origin in both of these patients. The 1st patient developed a mycotic pseudoaneurysm of the ascending aorta at the cannulation site, secondary to the development of Pseudomonas mediastinitis following aortic valve replacement. This sequela was successfully treated by means of aneurysmectomy and closure of the aorta with a bovine pericardial patch, under cardiopulmonary bypass with circulatory arrest. The 2nd patient developed pseudoaneurysm and perforation of the aorta at the cardioplegia needle site, secondary to Pseudomonas mediastinitis following aortic and mitral valve replacement. This patient died. In both patients, the cannulation site and the cardioplegia needle site had been closed with pledgeted sutures. Pseudomonas aeruginosa was cultured from both sites. Once the diagnosis of Pseudomonas mediastinitis is made following heart surgery, the patient should undergo reoperation, if possible, for removal of the foreign bodies (pledgeted sutures). In addition, these patients should be monitored with chest magnetic resonance angiography every 3 months for 1 year, in order to diagnose early development of a mycotic pseudoaneurysm and subsequent complications.

Aged↗

PubMed1986

Antigenic specificities of Pseudomonas aeruginosa alkaline protease and elastase defined by human T cell clones.

Virulent strains of Pseudomonas aeruginosa derive their pathogenicity, in part, from their secretion of two proteolytic enzymes, alkaline protease (AP) and elastase (E). Human T lymphocytes specific for AP and E can be detected in the blood of immune donors and have afforded the opportunity to characterize the antigenicity of these proteins. To accomplish this goal, we have recently selected 68 human T cell clones from five different Pseudomonas-immune donors and determined their fine specificities. Fifty-five (81%) were found to be protease specific, demonstrating the immunogenicity of the exoenzymes in humans. These clones defined five AP and three E specificities and suggested the existence of at least five allomorphic determinants expressed on the proteases of various Pseudomonas strains. Limiting dilution analysis confirmed a number of antigenic relationships suggested by the long-term T cell clones and revealed that T cells specific for allomorphic protease determinants were at least as frequent in the blood of immune donors as were T cells committed to conserved determinants. Thus, both primary and long-term human T cell clones showed specificity patterns that distinguished proteases from different Pseudomonas strains. These observations describe a heretofore unknown antigenic system of Pseudomonas aeruginosa that should assist in defining the nature and specificity of Pseudomonas immunity in humans.

Antigens, Bacterial↗