Search PubMed⌕ Search

Biomedical subjects

P E Coudron

Publications and source records attributed to P E Coudron.

28 records · Page 2Linked to original sources

Comparison of rapid urease tests, staining techniques, and growth on different solid media for detection of Campylobacter pylori.

Thirty-nine single antral biopsies (phase 1) and 99 sets of six antral biopsies (phase 2) were collected from 132 patients, and 87 (63%) yielded positive cultures for Campylobacter pylori. Of several primary media tested in phase 1, tryptic soy agar and Skirrow agar, each supplemented with 10% whole sheep blood, supported relatively good growth of C. pylori. In phase 2, four of the six biopsies in each set were tested with different urease systems. Selective urea agar for rapid identification was the most sensitive (39 of 63 [62%] at 1 h) and specific (100%); however, the difference between this system and the CLOtest was not statistically significant. The remaining two biopsies, one transported in saline and the other transported in a supplemented tryptic soy broth, were ground separately and inoculated onto tryptic soy agar and Skirrow agar, each supplemented with 10% whole sheep blood. In selected instances, 10% horse serum or 10% horse serum and 5 mM urea or 1% cholesterol were also added to the media. Smears stained with a modified Gram stain or acridine orange detected 68% of 63 culture-positive biopsies; no false-positive results were reported. Skirrow agar supported better growth of C. pylori than did tryptic soy agar; the nonselective medium was also overgrown with contaminants in 25 to 30% of the positive cultures. Based on colony size, Skirrow agar supplemented with 10% whole sheep blood, 10% horse serum, and 1% cholesterol supported optimal growth of C. pylori. Fresh media supported better growth than did prepoured commercial media (P less than or equal to 0.004). Saline was a convenient and satisfactory transport medium for antral biopsies.

Acridine Orange↗

Disseminated coccidioidomycosis. Unusual manifestations in a cardiac transplantation patient.

Because of the immunosuppressive therapy received by patients undergoing cardiac transplantation, disseminated infections, including disseminated fungal infections, often develop. Disseminated coccidioidomycosis developed in a 23-year-old man soon after undergoing orthotopic cardiac transplantation. Clinical manifestations included an unusual rash, severe myositis and arthropathy, a rapid downhill course, and pathologic evidence of widespread fungal invasion, including invasion of the cardiac graft. Detailed travel and geographic histories, and perhaps skin testing and antibody determinations for geographic-specific pathogens, should be part of the preoperative evaluation of all transplant candidates.

Adult↗

In-vitro activity of LY146032 against Staphylococcus aureus and S. epidermidis.

The antibacterial activity of LY146032, a new cyclic polypeptide, was compared with that of vancomycin against 163 isolates of methicillin-susceptible and methicillin-resistant Staphylococcus aureus and S. epidermidis. The MICs of LY146032 and vancomycin for 90% of the isolates were 1.0 mg/l and 2.0-4.0 mg/l, respectively. Interpretative guidelines for 15 micrograms and 30 micrograms LY146032 discs could not be established because all isolates tested were susceptible to LY146032, on the basis of achievable serum levels. With one exception, MBC to MIC ratios of LY146032 and vancomycin were less than or equal to 2. Inoculum size had minimal effect on ratios. In time-kill studies, both drugs were 99.9% bactericidal at 6 h for four isolates at concentrations four times the MBC or less. No change in LY146032 activity was observed between isogenic pairs that were resistant to other antibiotics. Thus, LY146032 was very active in vitro against all staphylococcal isolates.

Anti-Bacterial Agents↗

Isolation of Stomatococcus mucilaginosus from drug user with endocarditis.

Stomatococcus mucilaginosus was isolated from the blood of a patient with endocarditis and a past history of drug abuse and aortic valve replacement. At autopsy, Gram stain of the aortic valve revealed gram-positive cocci. Our isolate was atypical for S. mucilaginosus in that colonies were nonmucoid and nonadherent to agar surfaces. Cellular capsules were demonstrated by light and electron microscopy. Phenotypic characteristics identified by conventional methods as well as profile numbers obtained by using two commercial identification systems for staphylococci, the API Staph-Ident and the dms Staph Trac, are presented. Practical tests that differentiate S. mucilaginosus from the genera Micrococcus and Staphylococcus include growth on nutrient agar containing salt and lysostaphin susceptibility. Additional tests that helped differentiate our isolate from group D streptococci included hydrolysis of L-pyrrolidonyl-beta-naphthylamide and streptococcal serogrouping.

Adult↗

Detection of Chlamydia trachomatis in genital specimens by the Microtrak direct specimen test.

The conventional cell culture method for detection of Chlamydia trachomatis requires two to six days and is technically difficult to perform. The authors evaluated a new, relatively simple, non-culture method (MicroTrak, Syva Co., Palo Alto, CA) that requires less than one hour to complete. Two hundred fifty-one cervical and 209 male urethral specimens from three Richmond health clinics were read by direct immunofluorescence staining and compared with cell culture technics using iodine staining. Patient specimens were applied directly onto microscope slides (8 mm well) and stained with a fluorescein-labeled monoclonal antibody. Slides were examined for 10-15 minutes at X1,000 using an AO epifluorescent microscope and were considered positive if five or more typical elementary bodies were seen. The sensitivity, specificity, positive and negative predictive values for the direct smear were 89%, 97%, 85%, and 98% for males, and 93%, 96%, 85%, and 98% for females, respectively. The rapid direct specimen test appears to be a satisfactory method for detecting chlamydia in male and female genital specimens.

Cervix Uteri↗

Evaluation of laboratory tests for detection of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.

Few studies evaluating susceptibility testing of methicillin-resistant staphylococci have included isolates of Staphylococcus epidermidis, a known pathogen in many types of serious infections. We tested 175 S. epidermidis and 95 Staphylococcus aureus isolates to determine the most sensitive procedures for detecting methicillin-resistant staphylococci. Reference procedures included agar dilution with methicillin and 4% NaCl in the agar and broth microdilution with methicillin and 2% NaCl in cation-supplemented Mueller-Hinton broth. After 24 h of incubation, the results from both methods correlated well and were within 1 log2 dilution for all isolates tested. Only one-half of all resistant isolates (92 of 183) were detected at 18 h by using the standard disk diffusion technique with 5-micrograms methicillin disks, and even fewer were detected with 10-micrograms methicillin disks and newly recommended zone-size criteria. However, the standard disk diffusion method with 4% NaCl in the agar increased the sensitivity and specificity for identification of the proper phenotype to greater than 92%. The spread plate and new spot techniques, both using agar with 4% NaCl, were also sensitive methods. Of 47 S. epidermidis isolates tested against oxacillin, 6 (13%) were oxacillin susceptible but methicillin resistant. Two automated systems, the Automicrobic system (Vitek Systems) and MicroScan (American MicroScan), as well as two broth screening systems available from Remel and Austin Biological Laboratories, failed to detect several resistant isolates, depending on the species.

Culture Media↗

Two similar but atypical strains of coryneform group A-4 isolated from patients with endophthalmitis.

Corynebacterium species and other coryneform organisms isolated from clinical specimens are frequently considered contaminants. We isolated two strains of a gram-positive organism from the vitreous fluid of two patients with endophthalmitis who had previously received intraocular lens transplants. The biochemical characteristics and gas chromatographic patterns of both isolates were similar to those of coryneform group A-4 strains. Major differences included esculin hydrolysis, nitrate reduction, growth pigment, and lactic acid production. These two strains along with a limited number of strains collected at the Special Bacterial Pathogens Laboratory (Division of Bacterial Diseases, Centers for Disease Control, Atlanta, Ga.) may represent a subgroup of coryneform group A-4. Results of in vitro susceptibility testing performed with antimicrobial agents commonly used to treat patients with bacterial endophthalmitis underscore the importance of determining MBCs for slow-growing organisms. This report cautions microbiologists not to discard organisms frequently considered contaminants when isolated from body fluids that are normally sterile and from patients receiving local steroids.

Actinomycetales↗

Streptococcus faecium outbreak in a neonatal intensive care unit.

An outbreak of bacteremia and meningitis in a neonatal intensive care unit is described. Seven cases occurred in premature infants with severe underlying diseases. An epidemiological investigation failed to document the reservoir of the epidemic strain but suggested that its transmission among the infants was via the hands of hospital personnel. All patients had nasogastric tubes and multiple intravascular devices, and the portal of entry may have been either the gastrointestinal tract or the sites of the intravascular devices. Conventional biotyping of isolates failed to differentiate between isolates from infected patients and isolates recovered from prevalence surveys and from the environment. However, rapid identification systems (API-20S [Analytab Products, Plainview, N.Y.] and the AutoMicrobic system [Vitek Systems, Inc., Hazelwood, Mo.]) were able to distinguish isolates recovered from infected patients and hands of hospital personnel from isolates recovered during prevalence and environmental surveys and 29 isolates from widespread geographical areas. This is the first known report of a nosocomial neonatal outbreak of bacteremia and meningitis due to Streptococcus faecium; it underscores the importance of identifying streptococci to species level.

Cross Infection↗

Chemical and catalytic properties of the peroxisomal acyl-coenzyme A oxidase from Candida tropicalis.

The peroxisomal acyl-CoA oxidase has been purified from extracts of the yeast Candida tropicalis grown with alkanes as the principal energy source. The enzyme has a molecular weight of 552,000 and a subunit molecular weight of 72,100. Using an experimentally determined molar extinction coefficient for the enzyme-bound flavin, a minimum molecular weight of 146,700 was determined. Based on these data, the oxidase contains eight perhaps identical subunits and four equivalents of FAD. No other beta-oxidation enzyme activities are detected in purified preparations of the oxidase. The oxidase flavin does not react with sulfite to form an N(5) flavin-sulfite complex. Photochemical reduction of the oxidase flavin yields a red semiquinone; however, the yield of semiquinone is strongly pH dependent. The yield of semiquinone is significantly reduced below pH 7.5. The flavin semiquinone can be further reduced to the hydroquinone. The behavior of the oxidase flavin during photoreduction and its reactivity toward sulfite are interpreted to reflect the interaction in the N(1)-C(2)O region of the flavin with a group on the protein which acts as a hydrogen-bond acceptor. Like the acyl-CoA dehydrogenases which catalyze the same transformation of acyl-CoA substrates, the oxidase is inactivated by the acetylenic substrate analog, 3-octynoyl-CoA, which acts as an active site-directed inhibitor.

Acyl-CoA Oxidase↗

Tetrazolium reduction as an aid for streptococcal growth detection with agar dilution susceptibility testing.

A dye reduction method for determining a definitive endpoint with agar dilution susceptibility testing has been developed. Bacterial growth was determined by applying to the inoculum spot a dye solution containing 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride and phenazine methosulfate. Viable colonies reduced the tetrazolium salt to a visible red color within 3 to 5 min. The minimum inhibitory concentrations of six antimicrobial agents tested against 167 clinical streptococcal isolates were recorded before and after the addition of the tetrazolium-phenazine methosulfate solution. A total of 252 discrepancies (25%) were observed, and of these, 30 (12%) differed by more than one tested antibiotic concentration. Endpoint reproducibility of the dye procedure was assessed by four technologists in a double-blind study. A 2.7-fold reduction in disagreement was observed when the dye was used. Use of the tetrazolium-phenazine methosulfate solution involves little deviation from standard antimicrobial susceptibility test procedures and yields more accurate, as well as reproducible, susceptibility results.

Agar↗