Relation of virulence of Escherichia coli in septicemia and urinary tract infection to temperature-sensitive growth in minimal medium.
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Biomedical subjects
Publications and source records attributed to L D Thrupp.
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This study evaluates a new direct rapid system for urine cultures, including detection and quantitation of positive specimens by Gram stain, direct identification by 4--6-hour incubation of sediment with reagent strips, and antibiotic susceptibility testing by direct (3--4-hour) disk-elution methods. Of 987 routine urine specimens, 121 had significant (less than or equal to 10(5) colony-forming units/ml) gram-negative bacilluria, of which 89% were detected by the Gram stain. Direct rapid identification was correct in 94%. Results of direct disk-elution antimicrobial tests showed overall agreement with results of standard disk diffusion of 93% of tests, and major discrepancies in 4%. For urine specimens with gram-negative bacilluria, this system permitted detection, quantitation, identification, and antimicrobial susceptibility testing in four to six hours with reasonable, though not complete, accuracy.
Intrauterine specimens were obtained from 22 patients with endometritis and 24 control patients following cesarean section by using both a new protected swab and a standard anaerobic swab. The protected swab improved the value of the direct smear and Gram stain, resulted in fewer false-positive cultures, better defined endometrial flora in patients with endometritis, and permitted major savings in laboratory personnel time and materials.
We have surveyed septicemic episodes occuring in individuals with acute leukemia and have found two factors favorably influencing survival, mainly granulocyte counts over 1,000 mm3 and gram-positive bacteremias. In contrast, blood cultures persistently positive for longer than 48 hours were a bad prognosticator. Significantly, patients with gram-positive bacteremia had received less antibiotics in the week prior to septicemia than had patients with gram-negative bacteremia.
To investigate the maternal-fetal transfer of tobramycin (TBM) and its distribution in the fetus, a single dose of 2 mg/kg was administered intramuscularly to 35 pregnant patients (13 first trimester, 22 second trimester) 0.5 to 34 h before hysterectomy. TBM concentration was assayed microbiologically in maternal serum, fetal tissues (placenta, brain, lung, liver, and kidney), and fluids (amniotic, cerebrospinal fluid [CSF], urine, and serum). Mean maternal serum half-life (1.54 h) and mean peak serum concentration of TBM were within ranges reported for nonpregnant adults. In fetal serum, half-life was 5.2 h, and TBM levels did not exceed 0.58 mug/ml. For intervals up to 34 h, the mean TBM concentration in placental tissues was 1.4 mug/g. Concentration differences related to fetal maturation were found for fetal CSF, amniotic fluid, and fetal kidney. No antimicrobial activity was found in the fetal CSF of >16 weeks' gestation. TBM was present predominantly in the second trimester amniotic fluid specimens. Fetal kidney concentrations reached 7.2 mug/g at 34 h after maternal drug administration. Higher TBM concentrations were related to advanced maturation of the fetal kidney. Second trimester fetal urine concentrations for TBM ranged from 0.1 to 3.4 mug/ml, and the fetal urinary half-life was 3.7 h. Knowledge of fetal pharmacology is essential for weighing the fetal benefits or risks of antimicrobial therapy for the infected gravid patient.
Infective valvular endocarditis, whether occurring on a native or prosthetic heart valve, continues to carry a serious prognosis. Death is usually due to congestive heart failure, arrhythmia, or embolic complications, rather than infection per se. While antimicrobial therapy and medications to control congestive heart failure continue to be the treatment of choice in most cases, early valve replacement is indicated in certain situations. During the past four years, 12 patients underwent valve replacement for infective endocarditis. Six patients underwent elective valve replacement after antibiotic or antifugal therapy. All survived the operation and were improved. Six other patients underwent emergency valve replacement. Two patients died intraoperatively and their operations, in retrospect, were delayed unnecessarily. Four patients survived and are improved. Prosthetic valve replacement during the course of infective valvular endocarditis should be considered based on indications such as congestive heart failure, rhythm disturbances, etc, rather than the status of the infection.
A two-phase collaboratice study was designed for definition of the extent to which results of disk diffusion tests can be reproduced. Two currently recommended techniques, the Kirby-Bauer method and the agar overlay method, were used. Of special concern were tests of Pseudomonas aeruginosa with moderate susceptibility to carbenicillin; gentamicin, tobramycin, polymyxin B, and colistin were also tested with P. aeruginosa as well as with Klebsiella pneumoniae, Escherichia coli, and Proteus species. The agar overlay method tended to give zones that were somewhat smaller than those given by the Kirby-Bauer method. Both disk methods demonstrated an excellent degree of reproducibility, being at least as reproducible as the standard agar dilution technique for measurement of minimal inhibitory concentrations. Slight changes in carbenicillin disk potency affected the zones of inhibition around P. aeruginosa more than those around susceptible E. coli. This fact suggests that a standard strain of P. aeruginosa would be advantageous for quality control of carbenicillin disk tests.
Stool specimens were examined from 40 children with diarrhea who were under three years of age to determine the incidence of enterotoxigenic Escherichia coli in endemic diarrhea. Heat-labile E. coli enterotoxin was assayed in the very sensitive and reproducible cultured adrenal tumor cell system. Toxigenic E. coli were isolated from only one stool specimen and in this case infection with Shigella dysenteriae was also present. None of the eight classic enteropathogenic E. coli isolates were positive in the adrenal assay. This study suggests that heat-labile enterotoxin-producing E. coli are not an important cause of endemic childhood diarrhea in Southern California.
Seven laboratories participated in a collaborative study to evaluate the Autobac 1 system. Results obtained with this assay system were compared to those obtained by the standardized Bauer-Kirby disk diffusion test, and each of these two methods was compared to the agar dilution technique. Comparison of the Autobac 1 and the disk diffusion results from the seven laboratories showed an overall average of 91.5% interpretive agreement with the 17 antimicrobial agents tested. The distribution in the levels of Autobac 1/disk diffusion agreement was such that with 13 antimicrobial drugs agreement was 90% or higher; with three, between 85 and 90%; and with one, 77% (nitrofurantoin). Comparison of the Autobac 1 and disk diffusion tests with the International Collaborative Study agar dilution test showed that both methods gave levels of agreement with the International Collaborative Study agar dilution technique that were generally high and equivalent. The average overall agreement between the agar dilution test and each of the other two methods was approximately 90%. Disagreements that did occur tended to involve organisms that were drug susceptible by the Autobac 1 system but intermediate or resistant by the other two methods. This was in part due to the narrow intermediate interpretive zone of the Autobac 1 test. In reproducibility studies with the Autobac 1 and disk diffusion methods, no significant differences were observed between the interpretive reproducibility of the two methods.
Tube dilution susceptibility tests in Trypticase soy broth showed that resistance to gentamicin (minimum bactericidal concentration >==12.5 mug/ml) among hospital isolates of Pseudomonas aeruginosa increased from 13.9% in 1969 to 38.9% in 1972. Transfer of drug resistance to six aminoglycosides from one wild Pseudomonas strain to another was accomplished in recombination experiments. A carbenicillin-resistant, beta-lactamase-producing strain served as the recipient. The exconjugant was resistant not only to aminoglycosides, including amikacin, but also to all clinically employed antimicrobials. Aminoglycoside resistance in the exconjugant was cured by sodium dodecyl sulfate. This transferable aminoglycoside resistance was not mediated by adenylylation or, as judged by bioassay, by other antibiotic-inactivating or -modifying processes.
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Standard methods for the methyl red (MR) test are not practical for routine use in clinical laboratories because of the necessarily prolonged incubation period. When read after overnight incubation, the usual MR test is often equivocal or falsely positive. The present study demonstrates the importance of standardizing the total volume of broth, the size of the vessel in which the cultures are incubated, and the density of the inoculum. In very small volumes of broth, cultures are better exposed to atmospheric oxygen, and thus MR-negative organisms tend to revert the initial acidic pH much more quickly than in deeper, large-volume broth cultures. In the proposed technique, a single colony was inoculated into a 0.5-ml amount of MR-Voges Proskauer (VP) broth (13- by 100-mm tube), and, after 18 to 24 hr at 37 C, one drop of MR was added. With this technique, the broth cultures produced either a definite red (positive) or yellow (negative) color, whereas various shades of orange were frequently observed when larger volumes of broth were tested after only 1 to 2 days of incubation. With 6.0-ml broth cultures, 18-hr MR tests were totally unreliable, but 18-hr tests in 0.5 ml of broth were comparable to the standard MR test performed after 5 days of incubation and superior to those performed after 48 hr in 6.0-ml broth cultures. With the proposed technique, the MR test can be incorporated readily into the routine scheme for identification of Enterobacteriaceae.
Infections with Klebsiella and Enterobacter have increased among hospitalized patients. To study such infections, relatively simple but precise methods are needed for clinical laboratories to identify the two genera accurately. Moreover, a rapid identification is essential for assisting with the therapy of the patients. For this purpose, a new 4-hr urease test was developed so that colonies could be tested directly from blood-agar plates which have been inoculated with clinical material and allowed to incubate overnight. This 4-hr test was positive with 98.5% of 202 Klebsiella species and negative with 80 Enterobacter species. As a single criterion for distinguishing between the two major genera, the new 4-hr urease test was just as accurate as a motility test (99% of the 282 isolates were accurately identified with either). The 4-hr urease test represents a simple, rapid, and reliable technique which is ideally suited for use in clinical microbiology laboratories.
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