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Pseudomonas mosselii sp. nov., a novel species isolated from clinical specimens.

Twenty-two fluorescent pseudomonad strains of clinical origin received as Pseudomonas fluorescens (10 strains), Pseudomonasputida (10 strains) and Pseudomonas sp. (2 strains), and 33 type strains of the genus Pseudomonas were studied by numerical analysis based on 280 phenotypic characters. Twelve of the 22 clinical isolates clustered within a specific group, cluster IV. The other strains clustered within groups containing well-characterized fluorescent Pseudomonas species or did not cluster. Strains belonging to cluster IV were phenotypically different from all other clusters and subclusters of fluorescent pseudomonads. DNA-DNA hybridization showed that cluster IV corresponded to a genomic group sharing 72-100% DNA relatedness. DNA-DNA hybridization values with 67 strains representing 30 species of the genus Pseudomonas sensu stricto, including six recently described species (Pseudomonas veronii, Pseudomonas rhodesiae, Pseudomonas libanensis, 'Pseudomonas orientalis', 'Pseudomonas cedrella' and Pseudomonas monteilii), were below 49%, the value found for P. monteilii. The DNA G+C content of the type strain was 63 mol%. Comparison of the 16S rRNA gene sequence of a representative strain of cluster IV (CFML 90-83T) with sequences of other strains of the genus Pseudomonas revealed that strain CFML 90-83T was part of the P. fluorescens intrageneric cluster. On the basis of phenotypic, DNA-DNA hybridization and phylogenetic analyses, a novel species, Pseudomonas mosselii sp. nov., is proposed for the 12 strains of cluster IV. The type strain is P. mosselii CFML 90-83T (= ATCC BAA-99T = CIP 105259T). The P. mosselii strains are phenotypically homogeneous and can be differentiated from other fluorescent species by several phenotypic features, including pyoverdine typing.

DNA, Bacterial↗

Pseudomonas grimontii sp. nov.

The vernacular name 'fluorescent Pseudomonas group 97-514' was coined for a group of 43 strains isolated from two French natural mineral waters. All these strains were gram-negative, rod-shaped and motile by means of a single polar flagellum. They produced fluorescent pigment (pyoverdin) on King B medium, catalase and cytochrome oxidase. They were capable of respiratory but not fermentative metabolism. They were not able to accumulate poly-beta-hydroxybutyrate and possessed an arginine dihydrolase system. DNA-DNA relatedness studies (S1 nuclease method) showed that the 43 strains of 'fluorescent Pseudomonas group 97-514' formed a genetically homogeneous group (DNA-DNA relatedness ranged from 70 to 100%). A total of 76 strains representing well-known or partially characterized species of the genus Pseudomonas sensu stricto had 7-56% DNA hybridization with strain CFML 97-514T. The highest DNA binding values were found with Pseudomonas veronii CIP 104663T (52%), Pseudomonas rhodesiae CIP 104664T (56%), Pseudomonas marginalis ATCC 10844T (56%), Pseudomonas gessardii CIP 105469T (53%) and Pseudomonas cedrella CIP 105541T (52%). Their unrelatedness was confirmed by deltaTm values greater than 7 degrees C. On the basis of the results of phenotypic and DNA-DNA hybridization studies, a novel Pseudomonas species, Pseudomonas grimontii sp. nov., is proposed for the 43 strains of 'fluorescent Pseudomonas group 97-514'. The type strain is strain CFML 97-514T (= CIP 106645T = ATCC BAA-140T). The G+C content of the DNA of the type strain was 58 mol%. A comparison of the complete 16S rRNA gene sequence of the type strain CFML 97-514T and the sequence of other strains of the genus Pseudomonas revealed that the novel species fell within the 'Pseudomonas fluorescens intrageneric cluster'. Members of P. grimontii grew at 4 degrees C but not at 41 degrees C. They were able to use D-xylose, alpha-L-rhamnose, alpha-aminobutyrate, meso-erythritol and itaconate as sole sources of carbon and energy and formed levan from sucrose. Strains do not possess lecithinase or Tween esterase activities. The clinical significance of P. grimontii is unknown.

DNA, Bacterial↗

High-performance liquid chromatography analyses of pyoverdin siderophores differentiate among phytopathogenic fluorescent Pseudomonas Species.

The relationship of pyoverdins produced by 41 pathovars of Pseudomonas syringae and by phytopathogenic Pseudomonas species was investigated. A high-performance liquid chromatography method for analyzing the culture medium proved to be superior to isoelectric focusing for detecting pyoverdin production, for differentiating slightly different pyoverdins, and for differentiating atypical from typical Fe(III)-chelated pyoverdins. Nonfluorescent strains were found in Pseudomonas amygdali, Pseudomonas meliae, Pseudomonas fuscovaginae, and P. syringae. Pseudomonas agarici and Pseudomonas marginalis produced typical pyoverdins. Among the arginine dihydrolase-negative fluorescent Pseudomonas species, spectral, amino acid, and mass spectrometry analyses underscored for the first time the clear similarities among the pyoverdins produced by related species. Within this group, the oxidase-negative species Pseudomonas viridiflava and Pseudomonas ficuserectae and the pathovars of P. syringae produced the same atypical pyoverdin, whereas the oxidase-positive species Pseudomonas cichorii produced a similar atypical pyoverdin that contained a glycine instead of a serine. The more distantly related species Pseudomonas asplenii and Pseudomonas fuscovaginae both produced a less similar atypical pyoverdin. The spectral characteristics of Fe(III)-chelated atypical pyoverdins at pH 7.0 were related to the presence of two beta-hydroxyaspartic acids as iron ligands, whereas in typical pyoverdins one of the ligands is always ornithine based. The peptide chain influenced the chelation of iron more in atypical pyoverdins. Our results demonstrated that there is relative pyoverdin conservation in the amino acids involved in iron chelation and that there is faster evolution of the other amino acids, highlighting the usefulness of pyoverdins in systematics and in identification.

Amino Acids↗

Passive immunity against pseudomonas sepsis during granulocytopenia.

Specific passive immunity against Pseudomonas aeruginosa sepsis was assessed in granulocytopenic dogs. Dogs were infused with either normal or antipseudomonas immune plasma 24 h before pseudomonas challenge. They were challenged intravenously with 10(7) serotype 6 P. aeruginosa during granulocytopenia. Treatment was evaluated by observation of survival periods, febrile responses, type 6 pseudomonas antibody titers, and quantitative cultures of blood and tissues. The results demonstrated that passively immunized dogs did not survive infection. Both normal-plasma and immune-plasma recipients had bacteremia at death, with median values of 980 and 470 pseudomonas per ml of blood, respectively. All dogs had marked febrile responses 24 h after pseudomonas challenge and had high concentrations of pseudomonas in their lung tissue at death, with median values of 10(8) pseudomonas per g of wet tissue weight. After plasma infusion, immune-plasma recipients had high concentrations of anti-pseudomonas antibody, with total antibody titers ranging from 256 to 1,024 and a median value of 1,024. These titers were comparable to titers attained in a previous study from our laboratory using active immunization with pseudomonas lipopolysaccharide vaccine, where the median total anti-pseudomonas antibody titer was 2,048. Actively immunized animals, however, were significantly protected against pseudomonas sepsis and had prolonged survival periods and prevention of bacteremia. The present study demonstrates that circulating type-specific antibody is not solely responsible for the protection afforded to granulocytopenic dogs actively immunized against pseudomonas.

Agranulocytosis↗

Pseudomonas infections in the thermally injured patient.

Pseudomonas aeruginosa, remains a serious cause of infection and septic mortality in burn patients, particularly when nosocomially acquired. A prototypic burn patient who developed serious nosocomially acquired Pseudomonas infection is described as an index case which initiated investigations and measures taken to identify the source of the infection. The effect of changes in wound care to avoid further nosocomial infections was measured to provide data on outcome and cost of care. The bacteriology of Pseudomonas is reviewed to increase the burn care providers understanding of the behaviour of this very common and serious pathogen in the burn care setting, before reviewing the approach to detection of the organism and treatment both medically and surgically. After controlling the nosocomial spread of Pseudomonas in our burn unit, we investigated the morbidity and mortality associated with nosocomial infection with an aminoglycoside resistant Pseudomonas and the associated costs compared to a group of case-matched control patients with similar severity of burn injury, that did not acquire resistant Pseudomonas during hospitalization at our institution. We found a significant increase in the mortality rate in the Pseudomonas group compared to controls. The morbidity in terms of length of stay, ventilator days, number of surgical procedures, and the amount of blood products used were all significantly higher in the Pseudomonas group compared to controls. Costs associated with antibiotic requirements were also significantly higher in the Pseudomonas group. Despite this increased resource consumption necessary to treat Pseudomonas infections, these efforts did not prevent significantly higher mortality rates when compared to control patients who avoided infection with the resistant organism. Thus, in addition to the specific measures required to identify and treat nosocomial Pseudomonas infections in burn patients, prevention of infection through modification of treatment protocols together with continuous infection control measures to afford early identification and eradication of nosocomial Pseudomonas infection are critical for cost-effective, successful burn care.

Adult↗

Application of polynomial models to predict growth of mixed cultures of Pseudomonas spp. and Listeria in meat.

Three models for one rapid and one slow growing strain of Pseudomonas fragi and one slow growing strain of P. fluorescens were developed in a meat broth; they were designed to take account of variations in growth and to provide a growth response interval. These models, and another for Listeria monocytogenes (Lm14 model), were used to predict the growth of spoilage Pseudomonas spp. and pathogenic Listeria in meat products. The Pseudomonas and Listeria models provided satisfactory predictions concerning inoculated strains grown in decontaminated beef meat. It was also possible to use the Pseudomonas models to predict the growth of the natural flora (mainly Pseudomonas spp.) of refrigerated meat stored under aerobic conditions. In experiments with mixed populations, three situations were observed: (1) in decontaminated meat, L. monocytogenes inoculated alone grew well at 6 degrees C, and this result was correctly predicted by the model; (2) in decontaminated meat inoculated with Listeria and Pseudomonas strains, L. innocua grew well and was not affected by the presence of Pseudomonas, and the growth of both organisms was correctly predicted by the models; (3) in naturally contaminated meat inoculated with Listeria, the strain did not grow until Pseudomonas had reached the stationary phase. The models satisfactorily predicted the growth of Pseudomonas spp. but not that of Listeria. In conclusion, the Lm14 model cannot be used for refrigerated meat stored aerobically as the results suggest a 'fail-safe' level which may be too high: meat had already reached a spoilage state even though no increase in the level of Listeria was observed. The Pseudomonas models accurately predicted the growth of naturally occurring Pseudomonas spp.

Animals↗

Conservation of the response regulator gene gacA in Pseudomonas species.

The response regulator gene gacA influences the production of several secondary metabolites in both pathogenic and beneficial Pseudomonas spp. In this study, we developed primers and a probe for the gacA gene of Pseudomonas species and sequenced a 425 bp fragment of gacA from ten Pseudomonas strains isolated from different plant-associated environments. Polymerase chain reaction analysis and Southern hybridization showed that gacA is highly conserved within the genus Pseudomonas: multiple strains of different Pseudomonas species all responded positively to the probe, whereas no response was obtained from 18 other strains representing 14 species that belong to eight different genera of Gram-negative bacteria other than Pseudomonas. Furthermore, from a total of approximately 550 indigenous bacterial isolates obtained from the rhizosphere of wheat, all isolates that hybridized with the gacA probe were classified as Pseudomonas spp. by group-specific primers. Isolates that did not respond with the gacA probe and primers were identified as bacterial genera other than Pseudomonas, including Stenotrophomonas, Cryseomonas and Comamonas spp. These results indicate that gacA can be used as a complementary genetic marker for detection of Pseudomonas spp. in environmental samples. Phylogenetic relationships inferred from the newly sequenced gacA fragments and the sequences of gacA homologues present in the databases, showed six distinct clusters that correspond to the following bacterial families: Pseudomonaceae, Enterobacteriaceae, Alteromonadaceae, Vibrionaceae, Burkholderia and Xanthomonas. Within the Pseudomonadaceae and Enterobacteriaceae, polymorphisms within gacA and its homologues allowed identification of six and five subclusters respectively. Comparison of the gacA gene and GacA protein-based trees with the tree inferred from 16S rDNA sequences yielded a similar overall clustering. These results suggest that gacA and its homologues may provide complementary markers for phylogenetic studies of Pseudomonas spp. and Gram-negative bacteria other than Pseudomonas.

Bacterial Proteins↗

Effects of Pseudomonas colonization on body composition and resting energy expenditure in children with cystic fibrosis.

BACKGROUND: To determine the extent and effects of increased metabolic demand represented by Pseudomonas colonization on body composition and resting energy expenditure in children with cystic fibrosis (CF). METHODS: The study comprised 18 stable children with CF, of whom 10 (6 male/4 female) were colonized with Pseudomonas species and 8 (4 male/4 female) were not. The groups were of similar age range and genotype. Measured resting energy expenditure (REE) was performed by open circuit indirect calorimetry and compared with predicted REE calculated from standard equations. Body composition was determined by dual-energy x-ray absorptiometry, including lean body mass (LBM), fat mass (FM), bone mineral density (BMD), and anterior-posterior spine density (APS); these were compared using z-scores. Routine pulmonary function testing assessed forced vital capacity, forced expiratory volume in 1 second (FEV1), and forced expiratory flow over middle half of vital capacity (FEF25% to 75%); these were compared as percent predicted. RESULTS: As expected, results of pulmonary function testing showed significant deterioration among the children colonized with Pseudomonas species when compared with the children who were not, while standard anthropometry showed no differences in weight, height, or weight-for-height percentile and respective z-scores. Although a trend of lower LBM was noted among the children colonized with Pseudomonas species, no significant differences were found between these children and children who were not colonized with Pseudomonas species when z-scores for LBM, FM, BMD, or APS were compared during body composition analysis. In addition, neither REE as kilocalories per day (kcal/d) nor REE expressed as a percent predicted by standard equations discriminated between subgroups of children colonized with Pseudomonas species and children who were not. However, metabolic demand, expressed as resting energy expenditure in kilocalories per kilogram (kcal/kg) of LBM (REE/LBM), revealed significant differences between children colonized with Pseudomonas species and children who were not (75.4+/-4.4 vs 58.6+/-2.9 kcal/kg, p < .05). CONCLUSIONS: The effect of Pseudomonas colonization on metabolic demand in children with CF can be accurately assessed by expressing resting energy expenditure as kilocalorie per kilogram of LBM, the active metabolic component of the body. The 50% increase in REE/LBM seen in the children colonized with Pseudomonas species represents the metabolic demand from the inflammatory burden and work of breathing resulting from the effects of the Pseudomonas colonization. The trend of a lower LBM in the children colonized with Pseudomonas species makes this finding even more dramatic.

Absorptiometry, Photon↗

Development of a diagnostic PCR assay that targets a heat-shock protein gene (groES) for detection of Pseudomonas spp. in cystic fibrosis patients.

Laboratory detection of Pseudomonas spp., in particular Pseudomonas aeruginosa, remains an important assay in the management of patients with cystic fibrosis (CF). As the groES and groEL genes of P. aeruginosa have now been cloned and their nucleotide sequences determined, the aim of this study was to develop a novel PCR assay for the detection of Pseudomonas spp. from patients with CF by employing conserved primer regions of the groE heat-shock protein domain gene. A PCR assay was designed that targeted a 536 bp region of the groE gene to detect Pseudomonas spp. PCR amplification of genomic DNA from extracted organisms generated an amplicon of the expected size (approx. 536 bp) for all P. aeruginosa (n = 60), Pseudomonas putida, Pseudomonas fluorescens and Pseudomonas stutzeri isolates examined, but did not produce a positive amplicon for several other genera and species that are commonly isolated from the sputum of CF patients. RFLP analysis of the amplicons of all P. aeruginosa isolates demonstrated a single RFLP type that consisted of three bands at approximately 80, 190 and 250 bp; direct sequencing of the amplicons demonstrated the presence of a single sequence type, indicating the highly conserved nature of this region. In addition, the assay successfully produced a positive signal from primary non-selective plates of three known P. aeruginosa culture-positive CF patients, but was unable to generate a signal in a further six CF patients who had no history of infection with P. aeruginosa or other Pseudomonas spp. This assay is recommended to detect the presence of Pseudomonas spp., including P. aeruginosa, from primary culture plates that originate from laboratory analysis of CF patients' sputum, particularly at review, in those patients with no previous history of Pseudomonas infection or those who appear to be transiently colonized by this organism. Employment of such molecular methodologies, in conjunction with routine clinical sputum cultures, may provide improved information on the microbial status of CF patients, which will aid clinicians in both optimum patient management in terms of antibiotic regimes and CF centre infection-control practices.

Adolescent↗

Protective properties and haemagglutinins in serum from humans and in serum from mice injected with a new polyvalent Pseudomonas vaccine.

Mice given single injections of a polyvalent pseudomonas vaccine produced anti-pseudomonas haemagglutinins against the 16 component immunogens of the multivalent vaccine. Mice passively immunized with sera from vaccinated mice were protected against lethal challenge by 8/10 strains of Ps. aeruginosa of homologous serotype. Protection by the serum was inversely proportional to the virulence of the challenge strains. Anti-pseudomonas haemagglutinins were always present in sera which passively protected mice against pseudomonas infection. Low levels of anti-pseudomonas haemagglutinins were present in some sera which failed to passively immunize mice against pseudomonas infection. Anti-pseudomonas haemagglutinins and antibodies involved in passive protection were mainly in the IgM fractions of mouse serum. Control human sera contained anti-pseudomonas haemagglutinins against most serotypes of Ps. aeruginosa. Sera from patients with burns contained high levels of anti-pseudomonas haemagglutinins against some but not all serotypes of Ps. aeruginosa. Sera from both controls and patients with burns passively protected mice against pseudomonas infection.

Agglutinins↗

Specific detection of Pseudomonas spp. in milk by fluorescence in situ hybridization using ribosomal RNA directed probes.

AIMS: Pseudomonas spp. are considered the most important milk spoilage organisms. Here we describe development of a fluorescence in situ hybridization (FISH) probe specific for detection and enumeration of Pseudomonas spp. in milk. METHODS AND RESULTS: 16S rRNA sequences were analysed to develop specific oligonucleotide probe for the genus Pseudomonas. Twenty different Pseudomonas spp. and 23 bacterial species from genera other than Pseudomonas (as negative controls) were tested. All tested Pseudomonas spp. yielded a positive FISH reaction, whereas negative controls showed no FISH reaction except for Burkholderia cepacia that showed a relatively weak FISH reaction. The FISH assay specifically stains Pseudomonas in milk when the milk contains a mixture of other bacterial species. The FISH assay takes 2 h and compares favourably with current culturing methods, which take a minimum of 48 h. Specificity of the probe was validated using polymerase chain reaction to selectively amplifying the Pseudomonas rDNA gene and sequencing the gene products. CONCLUSIONS: The method presented in this study allows simultaneously detection, identification and enumeration of Pseudomonas spp. in milk. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid and accurate enumeration of Pseudomonas facilitates the identification of specific contamination sources in dairy plants, the accurate validation of pasteurization treatments and the prediction of shelf life of processed milk.

Animals↗

Isolation of a 48.5-kDa membrane protein from Pseudomonas maltophilia which exhibits immunologic cross-reaction to the beta-subunit of human chorionic gonadotropin.

In separate studies we have shown that Pseudomonas maltophilia (American Type Culture Collection 13637) possesses an immunoglobulin Fc-binding protein. We have found that this protein prevents the application of immunoassays using monoclonal antibodies to study possible production of a CG-like material by this bacteria. Employing an immunoglobulin saturation technique to block this protein as well as a zwitterion detergent membrane solubilization technique, we now report the isolation of a membrane protein from Pseudomonas maltophilia which shows immunological relationships to the beta-subunit and carboxyl tail of human pregnancy CG. This pseudomonas immunoreactive material produced dose-response curves in the following CG immunoassays: 1) a polyclonal rabbit anti-CG equilibrium assay, 2) carboxyl tail CG equilibrium assay, and 3) two CG equilibrium assays using monoclonal antibodies. The pseudomonas CG-like protein did not react in equilibrium assays for human TSH, human LH, or free alpha-subunit of CG. The pseudomonas CG-like protein was purified by affinity chromatography. The purified protein showed only 0.25% cross-reaction with pregnancy CG in the monoclonal antibody equilibrium assays. Furthermore, the purified protein showed no binding to rat testicular CG/LH receptors, but showed avid binding to the pseudomonas CG-binding protein previously described by Richert and Ryan. The pseudomonas protein showed no binding to Concanavalin-A, which avidly binds pregnancy CG. When assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, this protein had a mol wt of 48,500 daltons, which is larger than the mol wt of the unglycosylated beta-subunit of pregnancy CG. We conclude that pseudomonas contain a protein that has partial homology to the beta-subunit of pregnancy CG. This material does not bind to mammalian CG receptors, but does bind to a pseudomonas CG-binding site. The latter suggests it has a function, as yet unknown, in pseudomonas.

Bacterial Proteins↗

Patterns and routes of tracheobronchial colonization in mechanically ventilated patients. The role of nutritional status in colonization of the lower airway by Pseudomonas species.

Tracheobronchial colonization by Gram-negative bacteria is common in mechanically ventilated patients. Pseudomonas sp are commonly isolated from the lower airways. We hypothesized that Pseudomonas sp would preferentially colonize the lower airway and would be more common in patients with poor nutritional status. We serially collected 75 pairs of upper and lower respiratory tract cultures from 14 patients treated with mechanical ventilation for at least one week, examined patterns of airway colonization and routes of bacterial entry for Pseudomonas sp and other enteric Gram-negative bacteria (EGNB), and related these findings to host-associated factors, including nutritional status. Pseudomonas sp were the most common species isolates taken from the lower airway, found in nine of 14 patients and in 41.3 percent of all cultures. In contrast to other EGNB, Pseudomonas sp were found significantly (p less than or equal to 0.05) more often in the tracheobronchial tree (31 of 75 cultures) than in the oropharynx (18 of 75 cultures). Primary colonization of the lower airway by Pseudomonas sp was found in four patients, while other EGNB never followed this pattern when subjects were studied with cultures taken every third day. A host-related factor related to lower airway colonization by Pseudomonas species was poor nutritional status, assessed by a multifactorial index (p less than or equal to 0.01). We conclude that in mechanically ventilated patients, Pseudomonas sp colonize the lower airway in a different pattern and by a different route from those of other EGNB. The findings that Pseudomonas sp preferentially colonize the tracheobronchial tree may be important for the design of strategies to prevent airway colonization. The recognition that poor nutritional status, a potentially modifiable host-related factor, favors lower airway growth of Pseudomonas sp suggests one direction for future infection-control efforts.

Adult↗

Influence of mucoid coating on clearance of Pseudomonas aeruginosa from lungs.

Pulmonary infection with mucoid strains of Pseudomonas aeruginosa in present in the majority of cystic fibrosis patients with chronic lung disease. It has been postulated that this mucoid coating may act to decrease lung clearance of Pseudomonas by limiting access of phagocytes, antibodies, and antibiotics to the bacteria. To determine whether mucoid coating of Pseudomonas might decrease intrapulmonary killing, groups of guinea pigs were infected with intrabronchial instillations of equivalent numbers of mucoid and nonmucoid Pseudomonas. For this study, mucoid strains of Pseudomonas were obtained from cystic fibrosis sputa and passaged on blood agar plates to obtain their nonmucoid revertants. Animals were then sacrificed at timed intervals after infection, and quantitative cultures were performed on lung homogenates. In all cases, mucoid challenge strains retained their mucoid morphology after passage in guinea pig lungs. No difference in killing of mucoid and nonmucoid Pseudomonas could be detected at 6, 24, or 48 h after lung infection. Further challenge studies used guinea pigs that were either prevaccinated with lipopolysaccharide P. aeruginosa vaccine or else treated with tobramycin sulfate after infection. Nonvaccinated or untreated controls had reduced intrapulmonary killing of Pseudomonas compared with vaccinees or treated groups (P < 0.02 and P < 0.01, respectively). However, there were no differences in pulmonary killing of mucoid and nonmucoid Pseudomonas in the presence of either specific antibodies or antibiotic. We conclude from these studies that mucoid coating of Pseudomonas does not selectively impede mechanisms of intrapulmonary killing in guinea pig lungs.

Animals↗

Pulmonary clearance of Pseudomonas aeruginosa in neutropenic mice. Effects of systemic immunization.

Systemic immunization with killed Pseudomonas improves pulmonary clearance of these bacteria in normal mice. To test the effects of active systemic immunization on early pulmonary clearance of Pseudomonas in neutropenic mice, Balb/c mice were intraperitoneally immunized with 10(8) Pseudomonas or injected with saline twice weekly for 2 wk. Ten days after the last immunization, immunized and control mice received tail vein injections with either nitrogen mustard or saline daily for 3 days. Two days after the last nitrogen mustard injection, mice were intrabronchially challenged with 10(6) or 10(4) live Pseudomonas and clearance was assessed 4 h later. With the higher inoculum, immunization (neutrophil-replete and neutropenic mice) augmented pulmonary clearance. The improvement in clearance with the higher inoculum was associated with a marked increase in bronchoalveolar lavage anti-Pseudomonas IgG. However, bacteria grew in the lungs of all neutropenic mice. Immunization did not alter pulmonary clearance of the lower inoculum of Pseudomonas, and bacterial growth occurred in the neutropenic mice. Gentamicin was administered to mice 2 h after intrabronchial challenge with 10(6) Pseudomonas to determine whether its effects were additive to immunization. Gentamicin improved pulmonary clearance effected by immunization only in the neutrophil-replete mice; immunization provided no additional benefit to gentamicin in the neutropenic mice. These data show that both neutrophils and immunization contribute to Pseudomonas clearance. Immunization improves pulmonary clearance of Pseudomonas in neutropenic mice with a sufficiently high inoculum, but bacterial growth still occurs. Gentamicin treatment is effective in eliminating bacteria in this model and immunization has no additional benefit.

Agranulocytosis↗

Allograft colonization and infections with pseudomonas in cystic fibrosis lung transplant recipients.

OBJECTIVE: To assess the incidence of pseudomonal infection, colonization, and inflammation in the allograft of lung transplant recipients with cystic fibrosis (CF) as compared with recipients with other end-stage lung disease. DESIGN: Retrospective review. SETTING: University medical center transplant service. PATIENTS: All patients with CF and chronic pseudomonal infection (n=62) and patients with nonseptic end-stage lung disease (n=52) receiving a double lung transplant between October 1983 and March 1996. RESULTS: Fifty lung transplant recipients with CF survived beyond postoperative day (POD) 15 and were subject to sequential bronchoscopy with BAL. Forty-four CF lung transplant recipients had Pseudomonas isolated from the allograft by median POD 15 as compared with 21 non-CF lung transplant recipients (p<0.001) with isolation at median POD 158 (p<0.0001). Thirteen CF lung transplant recipients had histologic evidence of infection when Pseudomonas was isolated as compared with only three of the non-CF lung transplant recipients (p<0.01). These infections occurred earlier in the CF lung transplant recipients (median POD 10 vs 261) (p<0.01). When compared with non-CF lung transplant recipients, CF lung transplant recipients with Pseudomonas isolated but without concomitant histologic infection (colonized) were demonstrated to have increased number of polymorphonuclear cells (PMNs) in the BAL fluid recovered from the allograft (17.66+/-24.94 x 10(6) cells vs 3.46+/-4.73 x 10(6)) (p<0.05). Non-CF lung transplant recipients who became colonized with Pseudomonas also had a greater number of PMNs recovered when compared with non-CF lung transplant recipients who did not have Pseudomonas (22.32+/-34.00 x 10(6) cells vs 0.21+/-0.18 x 10(6)) (p<0.01). Nine of 32 (28%) lung transplant recipients with CF have died from pseudomonal allograft infections, but this is no greater than 4 of 21 (19%) deaths related to Pseudomonas infection in recipients without CF (p=0.34). CONCLUSIONS: Isolation of Pseudomonas from the lung allograft occurs more frequently and earlier after transplantation in recipients with CF. While infections related to Pseudomonas also occur more frequently in recipients with CF, there is no increase in mortality. There is an intense inflammatory response in the lung allograft associated with the isolation of Pseudomonas in recipients with and without CF.

Adult↗

[Taxonomic analysis of Pseudomonas strains with uncertain taxonomic status].

Pseudomonas strains isolated from soil and rhizosphere and designated as "Pseudomonas rathonis", Pseudomonas sp. B and Pseudomonas sp. C have been characterized by 113 phenotypic properties and studied by methods of molecular-genetic analysis. The 5'-terminal hypervariable 16S rRNA regions, containing from 208 to 344 nucleotides have been amplified and sequenced. The comparative analysis of sequence results gave evidence about phylogenetic relatedness of studied bacteria (98-100% of 16S rRNA sequences identity) to some saprophytic and plant-pathogenic Pseudomonas species. The latter substantially differed from studied strains in phenotypic characteristics. The DNA-DNA hybridization results have shown the low (0-36%) level of chromosomic DNA homology of "Pseudomonas rathonis", Pseudomonas sp. B, Pseudomonas sp. C strains with the type strains of P. fluorescens, P. putida, P. mendocina, P. pseudoalcaligenes. Obtained data give evidence that these strains belong to some novel Pseudomonas species.

Base Sequence↗

[Pulmonary Pseudomonas infections].

145 strains of pathogenic pseudomonas had been isolated from the sputum or bronchoscopic aspirate of 1423 patients with pulmonary infections. They were classified into 8 types, among which 31.7% was pseudomonas aeruginosa. Pseudomonas aeruginosa was the dominant causative organism in pulmonary infections of the aged while in presenile ones the organism was mainly Pseudomonas fetid. The incidence of nosnocomial pseudomonas infection in patients of COPD with respiratory failure was 40%, of COPD with pulmonary infection 9.1% and of others 6.6%. 24 pseudomonas carriers with COPD (colonies less than or equal to 10(6)/ml in sputum) had been followed up. 16 out of them became negative in sputum culture without any treatment, while the remaining 8 developed pulmonary pseudomonas infections. 21 patients (14.5%) were found to have other types of pseudomonas infections during antibiotic treatment. Sensitivity tests showed that third-generation cephalosporins and aminoglycosides had definite antimicrobial activity against pseudomonas, the former being more stable and effective than the latter.

Adult↗