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Combined pre-immunization and granulocyte transfusion therapy for treatment of pseudomonas septicemia in neutropenic dogs.

An experimental model was designed to evaluate a combined protocol of active immunization and granulocyte transfusions for treatment of Pseudomonas aeruginosa sepsis in the neutropenic host. One member of a pair of dogs was immunized with P. aeruginosa vaccine. Both dogs were then rendered transiently neutropenic with a single intravenous dose of cyclophosphamide (40 mg. per kilogram) and challenged with an intravenous inoculum of P. aeruginosa. Twenty-four and 48 hours after pseudomonas challenge each animal received granulocyte transfusions. Effectiveness of therapy was evaluated by observation of survival time, febrile response, and quantitative blood cultures. Results showed a significant increase in the survival period (P is less than 0.05), a lower febrile response (P is less than 0.025), negative blood cultures, and a greater recovery rate in the immune group. Immune dogs that died had negative blood cultures or less than or equal to 10 pseudomonas per milliliter of blood despite the presence of P. aeruginosa in tissues. In contrast, control dogs had septic deaths within 67 hours of pseudomonas challenge, marked febrile responses with 24 hours of infection, and positive blood cultures with 4,000 to 25,800 pseudomonas per milliliter of blood. These data show that combined therapy with immunization and granulocyte transfusions is effective in reducing the severity of P. aeruginosa infection and in preventing bacteremia during periods of leukopenia.

Agranulocytosis↗

DNA fingerprints of Pseudomonas spp. using rotating field electrophoresis.

Rotating field electrophoresis (RFE) was applied to evaluate the usefulness of this technique for identification of several Pseudomonas strains with suspected importance in deliberate releases. Genomes of common wild-type or genetically modified strains of Pseudomonas fluorescens, Pseudomonas stutzeri, Pseudomonas putida and Pseudomonas aeruginosa were digested with rare-cutting restriction endonucleases and subjected to pulsed field gel electrophoresis. Restrictions with SpeI or XbaI produced 11-28 large DNA fragments in the range of 50-500 kb pairs. The specific genomic fingerprints were different for most strains of the same species, but identical for closely related strains. Differences were not affected by the presence of natural or genetically modified plasmids.

DNA Fingerprinting↗

[Plasmid fingerprinting technique analysis of Pseudomonas aeruginosa isolated from a burn unit].

We used plasmid fingerprinting technique to analyse 50 strains of Pseudomonas aeruginosa isolated from our burn unit and 15 strains of Pseudomonas aeruginosa from other wards. The results revealed that the carrier rate of R-plasmid was 100%. All 50 strains of Pseudomonas aeruginosa from burn units contained one plasmid with approximate molecular weight of 55 kb. Among 15 strains of Pseudomonas aeruginosa from other wards, 10 strains contained one plasmid with the same molecular weight. However, 3 strains were found to possess an additional 12.5-kd plasmid and 2 strains contained 77-kb plasmid. This proves that R-plasmid carrying Pseudomonas aeruginosa is prevalent in the burn unit, and R-plasmid is disseminated in the hospital.

Burns↗

[Detection of oprI gene of Pseudomonas aeruginosa by reverse dot-blot hybridization].

OBJECTIVE: To establish a rapid method for detecting Pseudomonas aeruginosa at the early stage of infection. METHODS: Specific primers were designed according to oprI gene sequence of Pseudomonas aeruginosa, and the specific probe was synthesized by PCR. After photosensitive biotin labeling of the bacterial DNA, reverse dot-blot hybridization was used to detect Pseudomonas aeruginosa. RESULTS: The probe synthesized was highly specific to Pseudomonas aeruginosa without cross reaction with other bacteria, viruses or fungi. The method was capable of detecting 100 ng bacteria DNA. CONCLUSION: Reverse dot-blot hybridization possesses the merits of speediness and specificity in the detection of Pseudomonas aeruginosa in the early stage of infection.

Bacterial Proteins↗

[Investigation on the drug resistance of Pseudomonas aeruginosa in our burn ward in the past 11 years].

OBJECTIVE: To analyze the use of antibiotics and the drug resistance of Pseudomonas aeruginosa in the burn ward of our hospital in the past 11 years, so as to optimize the use of antibiotics in the future. METHODS: Bacterial epidemiology during 1991-2001 in our burn ward was investigated. The change of the drug resistance of Pseudomonas aeruginosa was observed by defined daily dose (DDD) of antibiotics in adult patients and by the ranking of antibiotic administration days. RESULTS: (1) Staphylococcus aureus (10.53%-34.40%) and Pseudomonas aeruginosa (75.66%-11.47%) were dominant in our burn ward. (2) Predominant antibiotics used included Penicillin, Amikacin, Vancomycin, Imipenem and Ceftazidime. (3) There was increasing drug resistance of Pseudomonas aeruginosa to the following antibiotics ranking in following order: Piperacillin (41.57%-100.00%), Imipenem (36.36%-98.46%), Ceftazidime (23.46%-97.85%), Amikacin (13.16%-100.00%) and ciprofloxacin (6.90%-100.00%). CONCLUSION: There was increasing drug resistance of Pseudomonas aeruginosa to all antibiotics, which might be related to antibiotic abuse.

Amikacin↗

[In vitro antimicrobial activity of defensins from rabbit neutrophils against Pseudomonas aeruginosa and its multiple-drug-resistance strains].

OBJECTIVE: To investigate the role and mechanism of defensins from rabbit neutrophils against the multi-drug-resistant (MDR) strains of Pseudomonas aeruginosa. METHODS: Defensins were extracted from rabbit peritoneal neutrophil granules with citric acid and purified by acid urea polyacrylamide gel electrophoresis (AU-PAGE). An agrose gel radial diffusion assay for bactericidal activity in vitro was used and ultrastructural changes of the killed Pseudomonas aeruginosa were examined by use of transmission electron microscope. RESULTS: NP2 was isolated and purified successfully by AU-PAGE. NP2 demonstrated concentration-dependent killing of Pseudomonas aeruginosa and its MDR strain. When the two kinds of Pseudomonas aeruginosa cells exposed to NP2 were examined by transmission electron microscope, we found that the cell membrane was incomplete, the components of cells condensed into pyknotic bodies accompanied by vacuolization, and there was some transudate between the cells. CONCLUSION: Defensin from the rabbit neutrophils exerts potent bactericidal activity against the MDR strain of Pseudomonas aeruginosa. The findings under electron microscope suggest the theory that defensins are capable of destabilizing the cell membrane permeability and causing the death of microorganism.

Animals↗

Characterization of Pseudomonas pathovars isolated from rosaceous fruit trees in East Algeria.

A survey of bacterial diseases due to Pseudomonas on rosaceous fruit trees was conducted. In forty two orchards located in the Constantine region ( East Algeria). Pseudomonas isolates were identified on the bases of their cultural and biochemical characteristics . A total of fifty nine phytopathogenic bacteria were isolated from diseased pome and stone fruit trees. Thirty one strains comparable to Pseudomonas syringae pv. syringae were isolated from cherry (Prunus avium L.), plum (P. domestica L.), apricot (P. armeniaca L.), almond (P. dulcis L.) and pear trees (Pirus communis L.); sixteen strains comparable to Pseudomonas syringae pv. morsprunorum were obtained from samples of cherry and plum. Twelve strains of Pseudomonas viridiflava were isolated from cherry, apricot and peach (Prunus persica L.).

Algeria↗

Study of the effect of Ammoides pusilla (Brot.) Breist, essential oil against Pseudomonas sp.

The economic losses caused by plant pathogens, the development of genetic resistance to applied pesticides by major pathogenic bacteria and public concerns about the use of synthetic chemicals on food and environment, increased interest in the search of alternative safe methods to control diseases spreading. Biological control with plant extracts has emerged as a promising option. The present study was carried out to determine the effect of Ammoides pusilla essential oil on the growth of two pathogenic bacteria, Pseudomonas syringae pv. syringae and Pseudomonas syringae pv. mosprunorum Three essential oil treatments, 1/2 v/v, 1/5 v/v and 1/10 v/v concentrations, were tested onto these bacteria, using the agar diffusion test. The results showed that 1/2 v/v concentration treatment completely inhibited Pseudomonas syringae pv. mosprunorum growth and produced a 35 mm diameter inhibition zone for Pseudomonas syringae pu. syringae. 1/5 v/v essential oil concentration produced inhibition zones of 19 and 17 mm diameter for Pseudomonas syringae pv. syringae and Pseudormonas syringae pv. mosprunorum respetively, while 1/10 v/v essential oil concentration gave inhibition plates of 11 and 12 mm diameter for both stains in the order cited. In in vitro culture essential oil gave promising results suggesting that future studies should be carried out and focus on determining appropriate formulation of this bio-molecule to be use as a bio-control agent.

Algeria↗

[Fatty acids composition of cellular lipids of the collected and newly isolated Pseudomonas lupini strains].

Fatty acid composition of cellular lipids of 23 Pseudomonas lupini strains (Beltjukova et Koroljova 1968) has been investigated. Cellular fatty acids which contained from C10 to C19 carbon atoms have been identified. Basic fatty acid of those Pseudomonas cells are hexadecanoic, hexadecenoic and octadecanoic acids. The 3-hydroxydecanoic (C10:0 3OH), 3-hydroxydodecanoic (C12:0 3OH), 2-hydroxydodecanoic (C12:0 2OH) and cyclopropane fatty acids which contain 17 and 19 carbon atoms have been detected in cellular lipids. The cellular fatty acids spectra of 22 P. lupini strains are similar to cellular fatty acids spectrum of the type strain Pseudomonas syringae pv. syringae 8511. Pathogenic isolate 2, which fatty acid content of cell lipids significantly differ from lipids of cell fatty acids from P. lupini strains and cell lipids of fatty acids of typical strains Pseudomonas syringae pv. syringae 8511 and Pseudomonas savastanoi pv. phaseolicola 9066 is the exception.

Carbohydrate Conformation↗

[Multiresistant Pseudomonas spp. in vitro susceptibility to a combination of two antibiotics].

INTRODUCTION: In vitro antibiotic combination testing would guide therapy selection in patients severely affected by multi-drug resistant Pseudomonas. OBJECTIVES: In vitro, a two-antibiotic combination susceptible against multi-drug resistant Pseudomonas isolated at the Laboratorio Clínico of the Hospital de Oncología, Centro Médico Nacional Siglo XXI in Mexico City were analyzed to determine which antibiotic combination showed the best bactericidal activity. MATERIAL AND METHODS: During 10 months, 30 multi-drug resistant Pseudomonas strains were tested. An automated method was used, including a diluting solution with a well-known concentration of a second antibiotic. Quality controls recommended by the NCCLS were used. Pseudomonas aeruginosa ATCC 27853; Escherichia coli ATCC 25922; and Escherichia coli ATCC 35218. Combinations were betalactamics-aminoglycosides; carbapenemis-amikacin; fluoroquinolones-cefepime; and ciprofloxacin-ampicillin. RESULTS: Ampicillin-ciprofloxacin combination was bactericidal against 100% of the isolates. Cefazolin, cefixime and ticarcillin with amikacin: <50%; aztreonam, cefoxilin, cefuroxime, cefotaxime, ceftazidime and piperacillin with amikacin: 50-60%; cefepime with gentamicin: 76%; cefepime with amikacin: 86%; imipenem and meropenem with amikacin: 70% and 76%; cefepime with ciprofloxacin: 83%; cefepime with levofloxacin: 73%. CONCLUSIONS: In vitro antibiotic combination susceptibilities against multi-drug resistant bacteria would be the only way to guide clinicians to select the best therapy in severe infections. We found that the ampicillin-ciprofloxacin combination showed the best in vitro effect against multi-drug resistant Pseudomonas.

Aminoglycosides↗

[Inhibition of adhesion to buccal epithelial cells of Pseudomonas aeruginosa by dextran].

Adhesion of Pseudomonas aeruginosa strains to buccal epithelial cells appears to be a necessary precondition for colonization and infection of respiratory tract. There are many strategies to prevent host organisms for Pseudomonas aeruginosa. The purpose of these studies was to evaluate the potential for preventing adhesion of Pseudomonas aeruginosa to epithelial cells with dextran. Dextran (5,000 MW) inhibited adhesion of Pseudomonas aeruginosa to buccal cells, at 20 mM was most inhibitory. The inhibitory effect appeared to be nonspecific because other neutral polysaccharides (glycogen and mannan) were also inhibitory. Dextran is an inexpansive and nontoxic agent and may be useful to prevent colonization and infection of respiratory tract with Pseudomonas aeruginosa.

Animals↗

[Study on serum-sensitivity of Pseudomonas aeruginosa].

Using a microtiter plate for cell culture one hundred strains of Pseudomonas aeruginosa isolated from various clinical materials were quantitatively investigated for sensitivity to the bactericidal activity of normal human serum. The sensitivity to the serum was widely distributed among these strains. In parallel, 100 strains of Pseudomonas aeruginosa were serotyped using a type-specific anti-Pseudomonas antiserum kit. All type M strains were proven to be serum-sensitive, while types G, B, and A were generally serum-resistant. Using an experimental model of osteomyelitis in mice, osteomyelitis could not be caused by serum-sensitive strains of Pseudomonas aeruginosa. These results would indicate that serum-resistance is an important factor in pathogenicity of Pseudomonas aeruginosa.

Adult↗

Pseudomonas aeruginosa bacteremia in patients undergoing liver transplantation: an emerging problem.

In our institution, Pseudomonas aeruginosa bacteremia appeared to occur with increasing frequency in patients undergoing liver transplantation. We thus conducted a prospective study to define risk factors and outcome in these patients. Over a 19-month period 6% of liver transplants were followed by Pseudomonas bacteremia. The mean age was 46 years (range, 24 to 67 years). The interval between transplantation and onset of bacteremia was 3 to 372 days (mean, 80). The incidence of Pseudomonas bacteremia in liver transplants was three times that of other transplants (heart, lung, kidney). Ninety one percent of infections were nosocomial. Polymicrobial bacteremia occurred in 30% of episodes. The portal of entry was respiratory in 30%, abdominal in 35%, and biliary in 13%. Four patients had recurrent Pseudomonas bacteremia: liver abscess (1), biliary obstruction (2), subhepatic abscess (1). Survival at 14 days was 70%. Survival rates were significantly lower for patients with hypotension, on mechanical ventilators, and increasing severity of illness (p less than 0.05). Survival was higher when bacteremia occurred within the first 30 days after transplantation compared to after 30 days. A large number (43.4%) of Pseudomonas bacteremias occurred after transplant surgery of biliary tract manipulation, while the patient was receiving a prophylactic regimen of cefotaxime and ampicillin. P. aeruginosa is an important pathogen in the liver transplant recipient; prevention may be possible for a subgroup of patients with the use of prophylactic antibiotics with activity against P. aeruginosa.

Adult↗

[Antagonistic action of Pseudomonas aeruginosa against Staphylococci, coliform bacilli and various types of Proteus].

Antagonistic activity of 2 fresh isolates and 3 collection strains of Pseudomonas aeruginosa against 177 microbial strains was determined with the method of late antagonism. Among the microbial strains there were 56 staphylococcal strains isolated from patents and carriers. 38 nontypable colon bacilli isolated from healthy persons, 59 enteropathogenic colon bacilli of various serogroups, 12 strains of Proteus and 12 colon bacilli, carriers of multiple drug resistance factors (R factors). All the cultures were sensitive to the antagonistic action of 5 or at least 3 strains of Pseudomonas used in the study. The most active antagonists were the fresh isolates of Pseudomonas as compared to the collection strains. Among the staphylococci S. aureus proved to be the most resistant to the antagonistic action of Pseudomonas as compared to S. epidermidis, the same as the strains isolated from carriers as compared to the strains isolated from patients. As for the enteric bacilli the most resistant were the strains of Proteus. Acquiring of transmissive R factors by the colon bacilli markedly increased their sensitivity to the antagonistic action of Pseudomonas.

Antibiosis↗

[Pseudomonas aeruginosa in raw and pasteurized milk].

Eighteen strains of Pseudomonas aeruginosa, i.e. 8.87%, were isolated during a year from 203 samples of raw milk. Two Pseudomonas aeruginosa strains, i.e. 4%, were isolated from 50 samples of pasteurized milk. The strains were isolated using propagation techniques in meat-peptone broth with malachite green and on selective media--on centrimide agar (CEM) and on Pseudomonas F agar. All the isolated strains produced protease, whereas lipase was produced by only five strains. The strains were devitalized when exposed to pasteurization temperatures (72 degrees C) for 20 seconds. At cold store temperatures (4 degrees C), Pseudomonas aeruginosa strain cells propagated on average by two orders, inhibitory effects of low temperatures were recorded only with one strain. Inhibitory effects of milk cultures (cream, yogurt) on Pseudomonas aeruginosa were observed; their effects were more clear-cut at the temperature of 4 degrees C. The strains were markedly susceptible to gentamycin.

Animals↗

[Method and result of oxygen addition in the culture process of Pseudomonas aeruginosa].

Pseudomonas aeruginosa septicemia are frequent complications of patients with major burns. To evaluate more rapid culture methods for diagnosis, we have established a new culture method of pseudomonas aeruginosa by oxygen addition. The results of vitro study showed that the growth time of pseudomonas aeruginosa was 8 to 12 hours earlier by the new culture method than by the ordinary method, when pseudomonas aeruginosa were incubated in broth at 37 degrees C to achieve approximately 50 to 65 colony-forming units/bottle. A total of 60 samples was tested when each culture bottle was added approximately 2 to 10 colony-forming units. The percentage of positive culture was 75% by venting oxygen addition method and 33.3% by ordinary method. The result of assay of 26 blood samples from patients with major burns showed that the percentage of positive blood cultures was 69.2% by the new method and 42.3% by the ordinary method. The culture time of the new method was 4 to 12 hours earlier than the ordinary method. This technique may lead to a greater chance of recovery and rapid detection of pseudomonas aeruginosa from blood samples.

Bacteriological Techniques↗

[Significance and problems of pseudomonas infection in ENT medicine].

Pseudomonas aeruginosa is frequently cultured in chronic otitis media, in external otitis, and from infected radical mastoid cavities. Necrotizing external otitis is a typical infection caused by Pseudomonas aeruginosa. Perichondritis of the external ear is sometimes due to a Pseudomonas aeruginosa infection. Some cases of chronic sinusitis and wound infections in patients with tracheostomy are caused by this organism. Antiseptic agents or astringents should be used for local treatment of Pseudomonas infected ears. If these fail local treatment with azlocillin for several days is a good alternative. Systemic antibiotic therapy is also available for Pseudomonas aeruginosa infections.

Anti-Bacterial Agents↗

Treatment and prevention of relapses of CAPD Pseudomonas peritonitis.

Pseudomonas peritonitis in continuous ambulatory peritoneal dialysis (CAPD) can be difficult to eradicate, because it is frequently resistant to common antibiotics, inducing the loss of the peritoneal cavity in some cases. A total of 14 episodes of Pseudomonas peritonitis in 12 patients (6 male, 6 female) were treated with intraperitoneal (IP) administration of a combination of ceftazidime and tobramycin. All patients were hospitalized. The loading doses were 1000 mg/2 L of ceftazidime and 1.7 mg/kg of tobramycin, and the maintenance IP doses were 250 mg/2 L of ceftazidime and 16 mg/2 L of tobramycin. The therapy duration was 14 days. In 7 episodes (group A) no other antibiotic regimen was provided, while in the remaining 7 episodes (group B) therapy was continued with 500 mg b.i.d. of oral ciprofloxacin for the next 14 days. Pseudomonas species isolated in group A were P. alcaligenis (1), P. putida (1), P. maltophilia (1), R. cepacia (1), and unidentified (3). In group B the following Pseudomonas species were isolated: P. aeruginosa (4), P. diminuta (1), P. stutszeri (1), and unidentified (1). Recurrence of peritonitis was seen in 4 episodes of group A with 2 catheter removals, while all episodes were cured in group B. These results suggest that IP ceftazidime and tobramycin with the additional use of oral ciprofloxacin is successful in the treatment and prevention of relapses of Pseudomonas peritonitis.

Ceftazidime↗