Biomedical subjects
Publications and source records attributed to T L Gavan.
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Because excessively high rates of false resistance have been encountered with the 10-mug amikacin disk in diffusion susceptibility tests, a study was performed to examine existing zone diameter interpretative criteria and to compare the accuracy of 10- and 30-mug amikacin disks by the error rate-bounded classification scheme. Although current zone diameter interpretative criteria eliminate false susceptibles, there is an unacceptably high rate of false resistants. This problem can be resolved in most instances by revising the zone diameter interpretative criteria for the 10-mug disk (resistant, </=9 mm; indeterminate, 10 to 11 mm; susceptible, >/=12 mm) or, preferably, by replacing the 10-mug disk with a 30-mug disk and adopting new interpretative criteria (resistant, </=14 mm; indeterminate, 15 to 16 mm; susceptible, >/=17 mm). Because of significant differences in performance among media, it is necessary to include Pseudomonas aeruginosa ATCC 27853 among controls routinely tested and to exclude from use lots of Mueller-Hinton agar yielding results outside the 75% tolerance (90% confidence) limits for amikacin.
The susceptibility of 11,840 clinical bacterial isolates to fortimicin A was determined by agar dilution or broth microdilution methods and compared with their susceptibility to amikacin and gentamicin. In general, the in vitro activity of fortimicin A was essentially the same as that of amikacin. Significant exceptions were the increased effectiveness of fortimicin A against Serratia marcescens and the greater activity of amikacin against Pseudomonas and other nonfermentative gram-negative bacilli. On a weight-for-weight basis, gentamicin showed greater activity than the other two antimicrobial drugs against most species; S. Marcescens was the major exception. However, at concentrations equivalent to achievable nontoxic serum levels, the proportion of isolates inhibited by the three drugs was quite comparable. There were several strains with unusually high resistance to one or more of the tested antibiotics. These usually occurred in one of the six participating institutions and could be traced to specific enzyme-producing or permeability mutants endemic to that particular institution.
More than 4,000 clinical urine specimens were evaluated with an automated microbial detection/identification system compared to a standarized manual analysis and the routine modalities used in five peer-group laboratories. The comparison indicates that the automated system recognizes the nine groups of significant microorganisms in urinary tract infections in hospitalized patients with the same efficiency as a standarized manual method. The automated system's ability to enumerate the bacterial populations in the original clinical specimen attained a high degree of accuracy.
The morphological and biochemical changes that occur after inoculation of sterile blood into a blood culture medium (tryptic soy broth) with sodium polyanetholesulfonate and CO(2) were investigated. Cellular changes, pH, PCO(2), and PO(2) were monitored and evaluated. Erythrocytes became crenated and developed precipitated hemoglobin inclusions within 4 h. The lymphocytes appeared morphologically intact at 24 h, and by 48 h a few cells had undergone transformation. Many neutrophils were vacuolated at 24 h. Neutrophils capable of phagocytizing Staphylococcus aureus were observed after 18 h of incubation. Identifiable eosinophiles were present on day 6 of the study. A decrease in PO(2) in the unvented bottles from 44.4 to 8 mm of Hg occurred by 24 h. PO(2) remained low for 6 days, after which a slight increase occurred. An increase in PO(2) in the vented bottle from 51 to 58 mm of Hg occurred by 24 h of incubation. In both the vented and unvented bottles the PCO(2) increased. This increase was markedly more rapid in the unvented bottle. From a pH of 7.06 a decrease occurred for the first 24 h after inoculation, with the pH stabilizing at 6.8 in the vented bottles and at 6.6 in the unvented bottles. The biochemical changes that occurred in the vented culture bottles stabilized more rapidly than those of the unvented bottles. Changes caused by the addition of sterile blood to a blood culture medium resulted in conditions which departed considerably from accepted optima for the isolation of clinically important microorganisms. The phagocytosis of organisms that occurred may also have reduced the yield.
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Reassessment of the "class" concept of disk susceptibility testing. Cephalothin disks versus minimal inhibitory concentrations with eleven cephalosporins. Am J Clin Pathol 70: 909--913, 1978. Studies were carried out to determine whether susceptibility or resistance to 11 cephalosporins could be predicted reliably from the results of tests with a single cephalothin disk. The cephalosporins were tested with a microdilution technic and with a standardized disk test. Strains susceptible to a cephalothin disk were predictably susceptible to all other cephalosporins. However, 2--12% of the strains were resistant to cephalothin disks but were susceptible to the more active parenteral drugs cefoxitin, cephamandole, cefuroxime, and BL-S786. Because of differences in antimicrobial activities, the cephalosporins could be divided into three subgroups for purposes of susceptibility testing: one subgroup includes the majority of cephalosporins and may be represented by tests with cephalothin, the second subgroup inclues three active parenteral drugs (cephamandole, cefuroxime, and BL-S786) and may be represented by tests with cefuroxime, and the third subgroup consists of cefoxitin, a cephamycin with a unique broad spectrum of activity. Until the drugs in the second and third subgroups are released for general therapeutic use, the practice of testing only one cephalosporin disk appears to be a reasonably reliable procedure.
Micro-Media Systems (MMS) has developed a procedure by which microdilution trays can be filled with dilutions of antimicrobial agents, frozen, and distributed to clinical laboratories. The trays are prepared in various distribution centers throughout the United States to supply clinical laboratories in the vicinity of each center. For use, trays are removed from the freezer, allowed to thaw, and then inoculated with inocula prepared as for any other susceptibility test, using a convenient disposable inoculator (ca. 5 mul per well). A collaborative study was planned to evaluate microdilution trays prepared in three Micro-Media Systems distribution centers. Microdilution minimal inhibitory concentrations (MICs) were compared to standard tube dilution tests (the international collaborative study group method). With gram-positive cocci, the two techniques gave essentially equivalent results. With gram-negative bacilli, the microdilution MICs were generally one doubling dilution lower than the standard tube dilution MICs. Similar results were seen with microdilution trays prepared with a Cooke Dynatech MIC 2000. Inter- and intralaboratory reproducibility with the macro- and microdilution techniques were quite satisfactory, i.e., at least 96% of the end points were within a range of +/-1 log(2) dilution intervals.
An automated and computerized system (Automicrobic System [AMS]) for the detection of frequently encountered bacteria in clinical urine specimens was tested in a collaborative study among six laboratories. The sensitivity, specificity, reliability, and reproducibility of the AMS were determined, and the system was compared with conventional detection and identification systems. In this study, pure cultures and mixtures of pure cultures were used to simulate clinical urine specimens. With pure cultures, the sensitivity of the AMS in identifying the nine groups of organisms most commonly found in urine averaged 92.8%. The specificity averaged 99.4%, and the reliability of a positive result averaged 92.1%. The latter value was strongly influenced by a relatively high occurrence of false positive Escherichia coli results. The AMS was capable of detecting growth of most organisms, including those which it was not designed to identify. However, it identified some of these incorrectly as common urinary tract flora. Reproducibility of results, both within laboratories and among different laboratories, was high. Fast-growing organisms, such as E. coli and Klebsiella/Enterobacter species, were detected often at cell populations well below the AMS enumeration threshold of 70,000/ml. In mixed culture studies, high levels of sensitivity and specificity were maintained but when Serratia species were present in mixtures with other organisms, there was often a false positive report of E. coli. The overall performance of the AMS was considered satisfactory under the test conditions used.
Laboratory microbiology provides to clinicians caring for patients with infectious diseases objective data regarding etiologic agent identification and antimicrobial susceptibility. Mutual communication is essential to ensure appropriate selection of materials for culture, prompt collection and transport, and efficacious handling in the laboratory. Training programs for physicians and clinical scientists are too few in number and are in need of expansion. Program in pathology and postdoctoral programs must emphasize clinical and laboratory experience so that communication can be improved.
Commercial systems designed and marketed for evaluation of Enterobacteriaceae are used in many clinical microbiology laboratories. Evaluations of these systems have been reported from several large laboratories. Identifications with one or more of these systems were compared with those obtained by conventional methods with many strains of organisms. The Microbiology Resource Committee of the College of American Pathologists designed two surveys to evaluate the performances of commercial systems with identical strains of organisms submitted to a large number of participants. The surveys used were the Special Bacteriology Survey D-D and the Comprehensive Microbiology Survey B-D submitted during 1975--1976. Single strains of Klebsiella pneumoniae and Serratia marcescens in pure culture were submitted in the Comprehensive Survey and the same strains of these organisms plus one strain each of Pseudomonas stuartii and Citrobacter freundii in the Special Survey. The data obtained from participants permitted comparisons to be made between the API-20E, Enterotube, and r/b systems. These comparisons included individual biochemical test results shared by at least two systems as well as the ability of each to identify the unknown organisms to genus alone and to both genus and species. The results are presented and subjected to statistical analysis.
Systemic absorption of neomycin sulfate occurred in ten patients in whom neomycin wound irrigating solutions were used during total hip replacement. The use of neomycin in this manner may be associated with the risk of ototoxicity and nephrotoxicity.
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Cefuroxime, a new parenteral cephalosporin was compared with cephalothin by broth microdilution susceptibility testing against 5,887 routine clinical bacterial isolates in four large clinical laboratories. The minimal inhibitory concentrations (MICs) of cefuroxime against the Enterobacteriaceae were consistently lower than those of cephalothin. This was most striking among the Enterobacter species, which were generally susceptible to cefuroxime (MIC </= 8 mug/ml), but resistant to cephalothin. Similar results occurred with Haemophilus species, Acinetobacter anitratus, meningococci, and Aeromonas hydrophilia, but Pseudomonas species and enterococci were resistant to high concentrations of both drugs. Streptococci showed slightly greater susceptibility to cefuroxime than to cephalothin. By contrast, staphylococci were more susceptible to cephalothin. Bacteroides fragilis was resistant to cefuroxime, but other anaerobes were generally susceptible.
Three separate hospital clinical microbiology laboratories using three different identification systems participated in the identification of Enterobactericeae from a central pool of 'unknown" clinical isolates. With conventional tubed media, API-20E (Anlytab Products Inc.) and R/B tube (Corning Diagnostics) systems, there was a 91.1% agreement in the species designation. No significant differences at the 95% confidence level were found among the systems. Evaluation of individual tests within the systems used revealed lysine decarboxylase of the conventional and citrate of the API-20E system to be significantly different from the same test within the other two systems. The lysine decarboxylase of the conventional system had species relatedness, whereas the differences in citrate of the API-20E system were not related to a particular species. These individual test variations did not affect final organism identification. Reproducibility, evaluated as the system's ability to designate the same identification on two separate occasions, was 92 to 94% for each system. Exact duplication of selected sets of reactions was 60% for conventional, 45% for API-20E, and 61% for R/B. The variations in sets of reactions differed with the system and with the organism involved. The findings suggest equivalency among the three systems in ability to identify common clinical isolates of Enterobacteriaceae and point out the limited usefulness of these systems for biochemical biotyping.
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