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Biomedical subjects

M B Coyle

Publications and source records attributed to M B Coyle.

51 records · Page 3Linked to original sources

Rapid antimicrobial susceptibility testing of isolates from blood cultures by direct inoculation and early reading of disk diffusion tests.

Disk diffusion tests, inoculated directly from positive blood cultures, were evaluated for accuracy of reading zone diameters after 4- and 6-h and overnight incubation. In comparisons with results from standard disk diffusion tests, the 4-h results were in agreement for 83% of tests with gram-positive organisms and 64% of tests with gram-negative organisms. When minor discrepancies were ignored, the 4-h readings were in agreement for 98% of the tests with gram-positive organisms and 95% of the tests with gram-negative organisms. After 6 h of incubation, 91% of the tests with gram-positive organisms and 86% of the tests with gram-negative organisms agreed with standard results. The agreement was 99% for tests with both gram-positive and gram-negative organisms when minor discrepancies were excluded. Very major discrepancies occurred in two tests (0.1%) with gram-positive organisms and were not observed in tests with gram-negative organisms. The frequencies of major discrepancies were 3.5% after 4 h, 0.6% after 6 h, and 0.7% after overnight incubation. Ampicillin and cephalothin tests with Escherichia coli and Klebsiella spp. accounted for 81% of the major discrepancies in tests with gram-negative organisms. Oxacillin tests accounted for more than half of the major discrepancies in tests with staphylococci. The results of this study, which did not include the newer antibiotics, indicate that direct susceptibility tests from blood cultures read after 6 h of incubation are more reliable than 4-h results and produce less than 1% major errors in comparisons with standard susceptibility tests.

Anti-Bacterial Agents↗

Detection of Cryptosporidium oocysts in sputum during screening for mycobacteria.

We report a case in which Cryptosporidium oocysts were detected during routine auramine screening of a sputum specimen for acid-fast bacilli. The patient was a 28-year-old male with acquired immune deficiency syndrome and intestinal cryptosporidiosis. Microbiology laboratory workers should be alert to the possibility that these oocysts may be present in sputum specimens from patients with acquired immune deficiency syndrome.

Acquired Immunodeficiency Syndrome↗

Campylobacter jejuni enteritis: efficacy of antimicrobial and antimotility drugs.

We analyzed retrospectively the illnesses of 82 patients with Campylobacter jejuni enteritis to ascertain the efficacy of antimicrobials and drugs that inhibit gastrointestinal motility. Forty-four patients were treated with only supportive measures consisting of diet modification and fluids; 22 others received an antimotility agent for at least 48 h; the remaining 16 were given an antimicrobial at or near the time of therapeutic intervention. The three groups were similar in terms of severity of symptoms and signs. There was a greater need for secondary antimicrobial therapy because of static or worsening illness in the group treated with antimotility agents (8/22, 36%) than in the others (4/44,9%; 2/16, 13%: p less than 0.02). Furthermore, six patients treated initially and 10 treated secondarily with erythromycin or tetracycline had illnesses of shorter duration than did untreated controls paired by age, sex, length and severity of symptoms and signs, hematochezia, and antimotility therapy (p less than 0.05). Thus treatment of C. jejuni enteritis with erythromycin or a tetracycline shortened the illness, but antimotility agents impeded the resolution of the infection.

Adolescent↗

Multiply antibiotic-resistant Staphylococcus aureus: introduction, transmission, and evolution of nosocomial infection.

A burn patient with a multiply antibiotic-resistant Staphylococcus aureus infection was transferred to Harborview Medical Center from a burn unit in another state. Despite standard wound precautions, transmission to 34 patients occurred during the subsequent 15 months. Twenty-seven of the patients were infected. Disease included pneumonia, empyema, bacteremia, endocarditis, osteomyelitis, and burn and wound infections. Seventeen of the 34 patients died. Phage typing and plasmid analysis showed the spread of multiply resistant S. aureus from the burn unit to the surgical intensive care unit where a study evaluating the use of chloramphenicol in cases of bowel sepsis was in progress. During this period the organism became resistant to chloramphenicol by acquiring either of two chloramphenicol R-plasmids. Using plasmid profiles and antibiograms, four epidemic strains were identified that assisted in identifying patient and personnel reservoirs. The outbreak was controlled only after rifampin was added to vancomycin treatment of infected patients, which correlated with eradication of the carrier state.

Adult↗

Erythromycin and clindamycin resistance in Corynebacterium diphtheriae from skin lesions.

Erythromycin- and clindamycin-resistant Corynebacterium diphtheriae isolates were recovered from skin lesions. Resistance to erythromycin and clindamycin was induced by a subinhibitory concentration (0.03 microgram/ml) of erythromycin. Clindamycin (0.07 microgram/ml) was a more effective inducer of its own resistance than of erythromycin resistance. Erythromycin-inducible cross-resistance to vernamycin B alpha was demonstrated in disk diffusion tests.

Clindamycin↗

Methods of measuring zones of inhibition with the Bauer-Kirby disk susceptibility test.

Standard Bauer-Kirby disk tests were performed with 85 selected isolates, each tested in triplicate by four different investigators. Each disk test was observed, and zone diameters were measured, under two lighting conditions (transmitted light and reflected light). The two lighting systems produced similar zone measurements (+/-2 mm) with 96% of the tests. When there were greater differences, zones appeared to be larger when observed with reflected light. Interlaboratory reproducibility was much greater when using reflected light rather than transmitted light. We concluded that zone diameters should be measured from the back of the plate while it is resting on, or held 2 to 3 inches [ca. 5.1 to 7.6 cm] above, a black, nonreflecting, flat surface, illuminated by a reflected light source.

Anti-Bacterial Agents↗

Simple inoculum standardizing system for antimicrobial disk susceptibility tests.

A collaborative study was undertaken to evaluate a simple, convenient device which expedites inoculum standardization for antimicrobial disk susceptibility tests. The Inocupac system (Minnesota Mining & Manufacturing [3M] Co., St. Paul, Minn.) was used to perform disk tests in parallel with the standard Bauer-Kirby method. Five investigators tested 100 selected isolates, each in triplicate. Inter- and intralaboratory precision of both disk procedures was essentially comparable. The Inocupac system failed to consistently provide satisfactory growth with some streptococci, but when testing common gram-negative bacilli, staphylococci, and most enterococci, the Inocupac system gave zones about the same size (+/-2 mm) as the Bauer-Kirby procedure. Interpretive agreement between the two test systems varied from 90 to 99% with different antimicrobial agents, and repeated tests with the Bauer-Kirby method demonstrated the same degree of interpretive agreement. The Inocupac system is a valid alternative method for inoculating disk susceptibility tests.

Anti-Bacterial Agents↗

Reproducibility of control strains for antibiotic susceptibility testing.

Inter- and intralaboratory reproducibility of susceptibility testing requires stable control strains. The Food and Drug Administration diffusion procedure recommends the Seattle strains of Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) for this purpose. It was of interest to determine the present reproducibility of control cultures maintained in various laboratories over several years. Fifteen cultures each of S. aureus and E. coli were obtained from laboratories in different parts of the country. Their performance was compared with strains directly derived from ATCC. Diffusion susceptibility tests using a modified overlay technique were made with four replicates. Seven of the eight statistically significant differences in responses of the staphylococci were to penicillin, methicillin, or cephalothin. One culture was a penicillinase producer with a zone 15 mm less than the standard strain. Eleven of the 15 cultures showed no significant deviations or differences greater than 2 mm from the results with the strain derived directly from ATCC. All except the penicillinase producer were of identical phage type. Among 150 organism-antibiotic combinations tested with E. coli, all but one reading were within 2 mm of the standard. Four of the six statistically significant differences were in a culture from one laboratory. The stability of the cultures appears to have been influenced by the method of storage. Cultures that were kept frozen during extended storage were remarkably stable. Significant differences were found in cultures from four of five laboratories that maintained cultures in refrigerators or at ambient temperature.

Escherichia coli↗

Genetic analysis of drug resistance in Neisseria gonorrhoeae: identification and linkage relationships of loci controlling drug resistance.

The genetic basis of multiple drug resistance of Neisseria gonorrhoeae was investigated by the technique of transformation. Six different genetic loci were characterized by the type and amount of antibiotic resistance they controlled, and also by the degree of linkage to other resistance markers. A streptomycin resistance locus is linked to separate loci determining resistance to tetracycline, chloramphenicol, and erythromycin. A multiple resistance locus was identified. This genetic locus determines resistance to a variety of antibacterial agents. Lastly, a locus determining resistance to the penicillins was found which is unlinked to any other resistance locus.

Chromosomes, Bacterial↗

Genetic analysis of drug resistance in Neisseria gonorrhoeae: production of increased resistance by the combination of two antibiotic resistance loci.

The studies reported here demonstrate that increased resistance of Neisseria gonorrhoeae to penicillin, tetracycline, and chloramphenicol results from the combined effect of two resistance loci. As shown by experiments with deoxyribonucleic acid from transformants carrying only a single resistance locus, transformants with an incresed level of resistance to penicillin result from the combination of a penicillin-specific locus, pen, and a multiple resistance locus, mtr. Similarly, transformants with an increased level of resistance to tetracycline result from the combination of mtr and a tetracycline-specific locus, tet. Transformants with an increased level of resistance to chloramphenicol result from the combination of mtr and a chloramphenicol-specific locus, cml. Deoxyribonucleic acid dilution experiments established that only a single dose of each of the two required resistance loci is necessary to give higher-level resistance. Higher-level-resistant transformants were not obtained when a double dose of one resistance locus or a combination of loci pairs other than mtr and pen, mtr and tet, or mtr and cml was introduced into a recipient. Combinations of the mtr and tet genes resulted in increased resistance to semisynthetic tetracyclines. The presence of the mtr and pen genes resulted in increased resistance to penicillinase-stable penicillins.

Anti-Bacterial Agents↗

Evaluation of four mycobacterial blood culture media: BACTEC 13A, Isolator/BACTEC 12B, Isolator/Middlebrook agar, and a biphasic medium.

Four commercially available mycobacterial blood culture systems were compared for sensitivity and time to detection of growth. A 5-ml volume of SPS-anticoagulated blood was cultured in a BACTEC 13A vial and a modified M7H11/BHI biphasic medium. In addition, two aliquots of Isolator concentrates, each derived from 5 ml of blood, were inoculated into a BACTEC 12B vial and onto a pair of Middlebrook 7H11 agar plates (M7H11). Mycobacteria were recovered from 32 of 180 cultured specimens (17.8%). Growth was detected in 30 (93.7%) of the 13A vials, 27 (84.4%) of the M7H11 agar plates, 26 (81.2%) of the 12B vials, and 14 (43.8%) of the biphasic bottles. The mean times to growth detection in the 13A vial (14.2 days) and the 12B vial (13.7 days) were shorter than in either the M7H11 plates (20.8 days) or the biphasic medium (24.1 days). When the Isolator/12B vial-and-M7H11 plates were evaluated as a single system, 29 cultures (90.6%) had a mean time to growth detection of 13.5 days. Colony-forming units per ml were inversely associated with time to growth detection. Delay in transport (greater than 24 h) appeared to reduce viability. The direct inoculation feature makes the 13A vial very suitable for mycobacterial blood cultures.

Culture Media↗