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J E Stout

Publications and source records attributed to J E Stout.

At least 37 records · Page 2Linked to original sources
PubMed1998

Comparative activity of ciprofloxacin, ofloxacin, levofloxacin, and erythromycin against Legionella species by broth microdilution and intracellular susceptibility testing in HL-60 cells.

Animal lung macrophages or human peripheral blood mononuclear cells have been used for testing intracellular activity of anti-Legionella antibiotics; such studies are labor intensive such that comparative antibiotic studies for the many Legionella species are few. We evaluated a human monocyte cell line (HL-60) as an alternative model. HL-60 (1.5 x 10(6) cells/well) was differentiated into adherent cell and infected with 1.5 x 10(7) CFU of Legionella pneumophilia (L. pneumophilia). Erythromycin and quinolones, ciprofloxacin, ofloxacin, and levofloxacin were added to cells at 1 and 8 x MIC. Percent (%) inhibition ratios equal to total L. pneumophila with agent divided by L. pneumophila without agent x 100 were determined at 48 h; lower ratios implied greater potency. By broth dilution in buffered yeast extract broth, the most potent agents against L. pneumophila were (MIC): ciprofloxacin (0.015-0.03), ofloxacin (0.015-0.03), levofloxacin (0.015-0.03), erythromycin (0.125-1.0 microgram/mL). In the intracellular model, the most potent inhibitors of L. pneumophila multiplication at 8 x MIC were (in order of potency) levofloxacin (24.2%), ciprofloxacin (30.6%), ofloxacin (37.1%), and erythromycin (55.0%). All the quinolones were highly active and significantly more potent against L. micdadei and L. bozemanii when compared to L. pneumophila.

Anti-Bacterial Agents↗
PubMed1998

Intermittent use of copper-silver ionization for Legionella control in water distribution systems: a potential option in buildings housing individuals at low risk of infection.

One copper-silver ionization system was sequentially installed onto the hot-water recirculation lines of two hospital buildings colonized with Legionella pneumophila, serogroup 1. A third building with the same water supply and also colonized with Legionella served as a control. Four weeks after activation of the system, distal site positivity for Legionella in the first test building dropped to zero. After operating for 16 weeks, the system was disconnected and installed onto the second test building. Twelve weeks of disinfection reduced the distal site positivity for Legionella in the second test building to zero. Legionella recolonization did not occur in the first test building for 6-12 weeks and in the second test building for 8-12 weeks after inactivation of the system. The control building remained Legionella-positive throughout the experimental period. A significantly higher copper concentration was found in the biofilm taken from a sampling device than in that from water. This is likely to be the reason that the copper-silver ionization system had the residual effect of preventing early recolonization. Our study raises the possibility that one copper-silver unit could be rotated among several buildings to maintain a Legionella-free environment. Such an approach may be cost-effective for buildings housing individuals at low risk for contracting legionnaires' disease.

Biofilms↗
PubMed1998

Activity of azithromycin, clarithromycin, roxithromycin, dirithromycin, quinupristin/dalfopristin and erythromycin against Legionella species by intracellular susceptibility testing in HL-60 cells.

We evaluated a human monocyte cell line (HL-60) as a model for testing the intracellular activity of anti-Legionella antibiotics; 1.5 x 10(6) HL-60 cells/well were differentiated into adherent cells and infected with 1.5 x 10(7) cfu of Legionella pneumophila. The most active agents against L. pneumophila as judged by broth dilution MICs were (in order of activity) azithromycin, clarithromycin, roxithromycin, quinupristin/dalfopristin, erythromycin and dirithromycin. The most active inhibitors of L. pneumophila intracellular multiplication were (in order of activity) azithromycin, erythromycin, quinupristin/dalfopristin, roxithromycin, dirithromycin and clarithromycin. All the agents were highly active against Legionella micdadei and Legionella bozemanii when compared with L. pneumophila.

Anti-Bacterial Agents↗
PubMed1998

Disinfection of water distribution systems for Legionella.

Hospital-acquired legionnaires' disease arises from the presence of Legionella in hospital water systems. Legionella not only persists in hot water tanks but is also found in the biofilm throughout the entire water distribution system. Conditions within water systems that promote Legionella colonization include water temperature, configuration and age of the hot water tank, physicochemical constituents of the water, plumbing materials, and commensal microflora. Hospital-acquired legionnaires' disease has been prevented by instituting control measures directed at the water distribution system. These include superheat-and-flush, copper/silver ionization, ultraviolet light, instantaneous heating systems, and hyperchlorination. Each of the above disinfection methods has been proven to be effective in the short-term, but long-term efficacy has been difficult due to limitations associated with each method. The complexities of Legionella disinfection, including advantages and disadvantages of each method, are reviewed. A successful Legionella prevention program requires cooperation and communication among hospital administrative personnel, engineers, and infection control staff. Routine environmental surveillance cultures for Legionella are the critical component for successful long-term disinfection. Culture results document the efficacy of the disinfection method and alert the hospital staff to consider Legionella in hospitalized patients with pneumonia.

Cross Infection↗
PubMed1998

Controlling Legionella in hospital water systems: experience with the superheat-and-flush method and copper-silver ionization.

OBJECTIVE: To evaluate the effect of copper-silver ionization on Legionella colonization and nosocomial legionnaires' disease and to compare the efficacy of metal ions versus the superheat-and-flush method of disinfection. DESIGN: Prospective determination over a 36-month period of copper and silver ion concentrations in the recirculating hot-water system, Legionella colonization of the hospital water distribution system, and cases of nosocomial legionnaires' disease. Retrospective comparison of results with the previous 13 years, during which the superheat-and-flush method was used. SETTING: The Pittsburgh Veterans' Affairs Health Care System (University Drive Division) acute-care hospital. INTERVENTION: Three copper-silver ionization systems were installed on the hot-water distribution system in November 1994. RESULTS: The average number of cases of legionnaires' disease per year and the percentage of distal sites positive for Legionella pneumophila for the superheat-and-flush method versus the copper-silver ionization method was six cases with 15% positivity versus two cases with 4% positivity, respectively. The reduction in Legionella colonization after copper-silver ionization was significant (P<.05) compared to the superheat and flush. Mean copper and silver ion concentrations (mg/L) were 0.29 and 0.054 from hot-water tanks, and 0.17 and 0.04 from distal outlets, respectively. CONCLUSIONS: We conclude that a properly maintained and monitored copper-silver ionization system was more effective than the superheat-and-flush method for reducing the recovery of Legionella from the hospital water distribution system.

Copper↗
PubMed1997

Legionellosis.

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Adult↗
PubMed1997

Multiply antibiotic-resistant gram-negative bacilli in a long-term-care facility: a case-control study of patient risk factors and prior antibiotic use.

OBJECTIVE: To determine the relation between prior exposure to specific antimicrobials and acquisition of gram-negative bacilli resistant to multiple beta-lactam and aminoglycoside antibiotics among long-term-care patients. DESIGN: Case-control study. Cases were patients from whom multiply resistant Enterobacteriaceae or Pseudomonas aeruginosa were isolated; controls were patients from whom nonresistant bacteria of the same species were isolated. Prospectively defined risk factors included underlying illness, activity level, presence of decubitus ulcers, presence of indwelling devices, and prior exposure to specific antimicrobial agents. Resistant and control isolates of P aeruginosa were compared using pulsed-field gel electrophoresis (PFGE) of genomic DNA after digestion with XbaI. SETTING: 390-bed long-term Veterans' Affairs facility. RESULTS: We identified 35 patients with multiply resistant Enterobacteriaceae and 24 patients with multiply resistant P aeruginosa. Of the resistant Enterobacteriaceae, 87% of isolates were resistant to piperacillin, 55% to ceftazidime, and 90% to gentamicin. Acquisition of multiply resistant Enterobacteriaceae was associated with presence of decubitus ulcers (odds ratio [OR], 12.2; 95% confidence interval [CI95], 3.3-44.2; P = .0002) and prior receipt of ampicillin (OR, 13.7; CI95, 2.2-84; P = .005). Of resistant isolates of P aeruginosa, 88% were resistant to piperacillin, 25% to ceftazidime, 42% to imipenem, and 67% to ciprofloxacin. Isolation of a multiply resistant P aeruginosa was associated with total days of antimicrobial exposure (OR, 1.07; CI95, 1.01-1.12; P = .011) and not with prior receipt of any individual agent. Eleven multiply resistant isolates shared a common PFGE pattern. CONCLUSIONS: In our long-term-care facility, acquisition of multiply resistant Enterobacteriaceae was associated with the presence of decubitus ulcers and prior exposure to ampicillin. Acquisition of resistant P aeruginosa was associated with total antibiotic exposure. Molecular typing of P aeruginosa isolates implicated patient-to-patient transmission of a limited number of resistant strains.

Aged↗
PubMed1996

Nosocomial Legionnaires' disease caused by Legionella pneumophila serogroup 5: laboratory and epidemiologic implications.

Environmental monitoring and clinical surveillance for Legionella species were done for 12 months as recommended by the Allegheny County Health Department (Pittsburgh). The water system of a hospital was found to be colonized with Legionella pneumophila serogroup 5. Three patients with nosocomial L. pneumophila serogroup 5 disease were subsequently diagnosed after laboratory tests for legionellae were made available for all patients with nosocomial pneumonia. All serogroup 5 isolates from the hospital water matched the 3 patient isolates by pulsed-field gel electrophoresis (PFGE). Furthermore, isolates found in the water supply dating back 10 years showed the same PFGE pattern. In contrast, 12 L. pneumophila serogroup 5 isolates from eight other institutions had different PFGE patterns. Routine environmental cultures were important in stimulating the application of Legionella laboratory testing, which subsequently identified unsuspected patients with nosocomial legionnaires' disease.

Aged↗
PubMed1996

Assessment of enzyme immunoassay versus radioimmunoassay for detection of Legionella pneumophila serogroup 1 antigen in frozen urine specimens.

An enzyme immunoassay (EIA) was compared with a radioimmunoassay (RIA) for detection of Legionella pneumophila serogroup 1 antigen in 37 frozen urine samples. Among 18 samples positive by the RIA at the time of collection, 50% (9 of 18) of stored specimens retained positivity when retested by the RIA whereas 78% (14 of 18) of stored specimens retained positivity when retested by the EIA (P < 0.02). A false-negative result occurred after storage for 5 months or less if the original RIA result showed weak positivity.

Humans↗
PubMed1995

Comparison of culture methods for monitoring Legionella species in hospital potable water systems and recommendations for standardization of such methods.

A lack of standardization of environmental monitoring techniques for Legionella spp. complicates the interpretation of results and comparisons of results from different institutions. A comparative assessment of techniques recommended by the Centers for Disease Control and Prevention, the Hygiene Institute (Graz, Austria), and our laboratory was performed. Variables investigated were sampling method (swabbing and collection of water samples [250 ml] before and after swabbing), method of concentration (none, filtration, and centrifugation), acid buffer treatment (no acid treatment, treatment for 3 min, and treatment for 15 min), and choice of medium (five formulations of buffered charcoal yeast extract agar with glycine, vancomycin, polymyxin B, anisomycin, or cycloheximide). Thirty-three sites in seven hospital buildings were studied. Recovery by swab correlated with recovery from water after swabbing (P < 0.05). However, the quantity of Legionella spp. recovered from swab specimens (mean, 3.0 x 10(4) CFU per swab) was greater than that recovered from water (mean, 4.7 x 10(3) CFU/250 ml). Filtration resulted in recovery rates (mean, 5.2 x 10(3) CFU/250 ml) higher than those by centrifugation (mean, 2.3 x 10(3) CFU/250 ml). Three minutes of acid buffer treatment to reduce overgrowth by commensal flora did not improve selectivity or sensitivity for Legionella spp. if glycine-containing selective media were used. Fifteen minutes of acid buffer treatment reduced recovery compared with that after a 3-min treatment. All glycine-containing media tested effectively inhibited background flora, but one selective medium containing dyes, glycine, vancomycin, and polymyxin B (DGVP) resulted in the greatest quantitative recovery of Legionella pneumophila. Use of buffered charcoal yeast extract agar and the acid buffer treatment gave the greatest recovery of non-pneumophila species. A standardized protocol with an emphasis on the culturing of swab samples is presented.

Bacteriological Techniques↗
PubMed1994

Controlled evaluation of copper-silver ionization in eradicating Legionella pneumophila from a hospital water distribution system.

A controlled evaluation was made of the efficacy of copper-silver ionization in eradicating Legionella pneumophila from a hospital water supply. Copper-silver ionization units were installed on the hot water recirculation line of one building with water fixtures positive for Legionella species. Another building with the same water supply served as a control. Legionella species persisted within the system when copper and silver concentrations were < 0.3 and < 0.03 ppm, respectively. When copper and silver concentrations were > 0.4 and > 0.04 ppm, respectively, there was a significant decrease in Legionella species colonization, but the percentage of water fixtures positive for organisms was unchanged in the control building. When the ionization unit was inactivated, water fixtures continued to be free of Legionella species for 2 additional months. Copper-silver ionization can eradicate L. pneumophila in a water distribution system. The advantages of copper-silver ionization include relatively low cost, straightforward installation, easy maintenance, nontoxic by-products and the presence of a disinfecting residual.

Copper↗
PubMed1994

Molecular fingerprinting of Legionella species by repetitive element PCR.

Repetitive element PCR (rep-PCR) uses outward-facing primers to amplify multiple segments of DNA located between conserved repeated sequences interspersed along the bacterial chromosome. Polymorphisms of rep-PCR amplification products can serve as strain-specific molecular fingerprints. Primers directed at the repetitive extragenic palindromic element were used to characterize isolates of Legionella pneumophila and other Legionella species. Substantial variation was seen among the rep-PCR fingerprints of different Legionella species and serogroups. More limited, but distinct, polymorphisms of the rep-PCR fingerprint were evident among epidemiologically unrelated isolates of L. pneumophila serogroup 1. Previously characterized Legionella isolates from nosocomial outbreaks were correctly clustered by this method. These results suggest the presence of repetitive extragenic palindromic-like elements within the genomes of members of the family Legionellaceae that can be used to discriminate between strains within a serogroup of L. pneumophila and between different Legionella species. rep-PCR appears to be a useful technique for the molecular fingerprinting of Legionella species.

DNA Fingerprinting↗
PubMed1993

Aspects of lung transplantation that contribute to increased severity of pneumonia. An experimental study.

In lung or heart-lung transplant recipients, complications as a result of pulmonary infections continue to be the most frequent causes of morbidity and mortality. This study was undertaken to identify the contributions of (1) thoracotomy, (2) interruption of lymphatic vessels and bronchial arteries, (3) transplant procedure, (4) drug-induced immunosuppression, and (5) graft allogenicity to the increased risk of pneumonia in lung transplantation. Lewis rats were inoculated with 10(5) colony-forming units of Legionella pneumophila serogroup 1 by direct instillation into the trachea after one of the following: a general anesthetic with no operation; a left thoracotomy; a left thoracotomy with pulmonary hilar stripping; an isogeneic orthotopic left lung transplant with or without immunosuppression; or an allogeneic transplant with immunosuppression with Brown-Norway rats as donors. Immunosuppression was induced with an intramuscular injection of cyclosporine (25 mg/kg of body weight) from the inoculation day to day 3. All rats were killed on day 6, and severity of infection was determined by quantitative culture of Legionella organisms in the lungs and spleen, titer of Legionella urinary antigen, differential cell count in bronchoalveolar lavage fluid, body weight loss, and gross inspection of the lung. Significant increases in lung Legionella concentration occurred as a result of the addition of pulmonary hilar stripping (from 10(5.13 +/- 0.34) in the thoracotomy group to 10(5.66 +/- 0.25) in the thoracotomy with hilar stripping group, p = 0.013) and the addition of immunosuppression (from 10(5.47 +/- 0.47) in the isogeneic transplant group to 10(6.94 +/- 0.52) in the isogeneic transplant with immunosuppression group, p = 0.00016). Thoracotomy, transplant procedures, and allogenicity itself resulted in no significant increases. The results for all other indicators paralleled those for lung culture. We conclude that the combination of drug-induced immunosuppression with lung denervation and interruption of lymphatic vessels and bronchial arteries results in the early development and increased severity of pneumonia in lung transplantation.

Animals↗
PubMed1992

Potable water as a cause of sporadic cases of community-acquired legionnaires' disease.

BACKGROUND: The environmental sources of sporadic, community-acquired legionnaires' disease are largely unknown, and culturing of water sources after identification of a case is currently not recommended. We conducted a prospective study of sporadic cases of community-acquired legionnaires' disease to determine whether the environmental reservoirs could be identified. METHODS: We cultured samples of potable water obtained from sources to which each of 20 patients with culture-confirmed, community-acquired legionnaires' disease had been exposed during the two weeks before the onset of symptoms. Monoclonal-antibody subtyping and restriction-endonuclease analysis were performed on the legionella isolates recovered from both the patients and the associated environmental cultures. RESULTS: For 8 of the 20 patients, isolates of Legionella pneumophila with identical subtypes were identified in cultures from both the patient and the potable water to which the patient had been exposed. The environmental reservoirs linked to the infections were the water supplies of two private residences, two nursing homes, two hospital outpatient clinics, and an industrial plant. CONCLUSIONS: Potable-water supplies that harbor L. pneumophila are an important source of community-acquired legionnaires' disease. Future studies should include attempts to identify the environmental sources of this infection.

Adult↗
PubMed1992

Cefamandole-susceptible strains of Legionella pneumophila serogroup 1: implications for diagnosis and utility as an epidemiological marker.

The standard selective Legionella medium that contains cefamandole failed to grow legionella pneumophila serogroup 1, subtype Bellingham, from a sputum sample from a patient with nosocomial Legionnaires' disease; the isolate did grow on a similar selective medium that substitutes vancomycin for cefamandole. Two Bellingham isolates from this patient's hospital environment also failed to grow when tested on the cefamandole medium. We tested 106 additional L. pneumophila serogroup 1 isolates that belonged to nine different monoclonal antibody subtypes and demonstrated that susceptibility to cefamandole was rare (10%) and limited to the Bellingham subtype. The diagnosis of Legionnaires' disease may be missed unless the culture protocol includes both a nonselective medium and a selective medium that does not contain cefamandole. In vitro susceptibility to cefamandole also provided an epidemiologic marker that linked a water source for a patient to nosocomial Legionnaires' disease.

Cefamandole↗
PubMed1992

Isolation of Legionella pneumophila serogroup 5 from empyema following esophageal perforation. Source of the organism and mode of transmission.

A patient undergoing esophageal dilatation for carcinoma of the esophagus suffered esophageal perforation and development of an empyema. Culture of pleural fluid yielded multiple organisms, including Legionella pneumophila serogroup 5. Epidemiologic investigation showed that the source of L pneumophila was a tap used by the nursing personnel to fill patients' water pitchers. Whole-cell restriction endonuclease analysis of DNA from the clinical and environmental isolates of L pneumophila serogroup 5 yielded identical patterns. Our findings suggest that L pneumophila was acquired by the patient at least 12 h prior to the procedure causing the esophageal perforation and empyema, suggesting that the organism can persist in an infectious form in the upper aerodigestive tract.

Aged↗