Prevention of Descending Pneumonia in Rats with Perflubron-delivered Tobramycin

OBJECTIVES: Patients undergoing emergent endotracheal intubation are at increased risk for developing pneumonia. Although numerous strategies have been investigated to reduce ventilator-associated pneumonia (VAP), the incidence of VAP and its associated mortality remains high. This investigation tested the hypothesis that LiquiVent (Alliance Pharmaceutical, San Diego, CA-LV) delivered antibiotics (via spray-dried microspheres-SDM) would improve survival in a rat model of descending gram-negative pneumonia. METHODS: Wistar rats (n = 49) were randomized to receive prophylaxis with 1). nothing (controls); 2). intramuscular (IM) tobramycin, 3). intratracheal LV plus SDM shells (vehicle), 4). intratracheal LV plus SDM shells plus IM tobramycin, or 5). intratracheal LV plus SDM containing 1 mg/kg of tobramycin. All interventions were given 24 hours before a bacterial challenge with 10(8) colony-forming units of intratracheal Klebsiella pneumoniae. Mortality at ten days was the sole outcome measure. Survival in individual groups was compared with controls by Fisher's exact test with Bonferroni correction for multiple comparisons. RESULTS: All animals in the control group died of pneumonia within ten days of bacterial inoculation (0% survival). Prophylaxis with either IM tobramycin or SDM vehicle plus IM tobramycin provided no protection (0% survival). This is in sharp contrast to the cohort receiving pretreatment with tobramycin-containing SDM delivered via LV, in which 60% of the animals survived to study completion (p < 0.05). CONCLUSIONS: Prophylaxis with SDM containing antibiotics delivered in low-dose LV provided significant protection in a rat model of descending gram-negative pneumonia. These data support the hypothesis that perfluorocarbon-delivered intratracheal antimicrobials may be useful in the prevention of VAP.     

Detection of beta-lactamase activity among clinical isolates of Branhamella catarrhalis with six different beta-lactamase assays.

A total of 74 different clinical isolates of Branhamella catarrhalis were examined for their ability to produce beta-lactamase by six different beta-lactamase assays. These included a conventional tube and disk test, in which the chromogenic cephalosporin nitrocefin was used as a substrate; a disk procedure, in which pyridinium-2-azo-p-dimethylanaline cephalosporin was used as a substrate; broth and disk acidometric methods; and a conventional tube iodometric assay. A total of 58 of the study isolates produced beta-lactamase. In all cases, positive results were obtained with the nitrocefin tube and disk assays after 1 min. With the pyridinium-2-azo-p-dimethylanaline cephalosporin disk test, 57 of the 58 beta-lactamase-producing strains yielded a positive reaction in 1 min; the remaining strain was positive after 10 min. None of the beta-lactamase-producing strains produced positive reactions by either the broth or disk acidometric methods after 1 min. With the broth test, 10 min was required for positive test results for 42 strains; 30 min was necessary for 16 strains. By the disk acidometric procedure, all 58 strains were positive after 10 min. Of 58 beta-lactamase-producing strains, 30 were positive by the iodometric assay after 1 min, 13 strains required 10 min, and 4 strains were detected as being beta-lactamase positive only after 30 min. One beta-lactamase-producing strain remained negative by the iodometric method. Among the 16 strains of B. catarrhalis that lacked beta-lactamase that were examined in this study, no false-positive results were obtained by any of the six assays.     

Neisseria sicca osteomyelitis.

Neisseria sicca was identified as the cause of vertebral osteomyelitis in a male patient who had previously suffered a nonpenetrating, traumatic back injury. The identifying characteristics and antimicrobial susceptibility patterns are presented for this rare human pathogen, which heretofore has not been reported as a cause of infection localized to bone.     

Branhamella (Neisseria) catarrhalis: criteria for laboratory identification.

Eleven clinically significant isolates of Branhamella catarrhalis grew well on modified Thayer-Martin medium and produced beta-lactamase, but did not grow on nutrient agar at 22 degrees C. Minimum inhibitory concentrations of vancomycin, colistin, and trimethoprim were found to be higher than the concentrations of these antibiotics in modified Thayer-Martin medium. The criteria necessary for laboratory identification of B. catarrhalis are discussed.     

Multicenter clinical laboratory evaluation of a beta-lactamase disk assay employing a novel chromogenic cephalosporin, S1.

S1, a new chromogenic cephalosporin (International BioClinical, Inc., Portland, Oreg.), was used to detect beta-lactamase production among a variety of commonly encountered bacteria in a four-center collaborative study. Results of an S1 disk assay were compared with those obtained by a nitrocefin-based disk procedure (Cefinase; Becton-Dickinson Microbiology Systems, Cockeysville, Md.), with repetitive testing of five quality control organisms and with individual tests of recent clinical isolates of Neisseria gonorrhoeae (162 strains), Haemophilus influenzae (162 strains), Moraxella catarrhalis (155 strains), Staphylococcus aureus (161 strains), and Bacteroides fragilis (164 strains). The performances of the two beta-lactamase disk assays were comparable for the first three species cited above. However, the S1 assay appeared to be a more sensitive procedure than the Cefinase assay when applied to S. aureus and B. fragilis, with respect to both total numbers of positive results and length of time to a definitive positive endpoint.     

Optimum use of selective plated media in primary processing of respiratory tract specimens from patients with cystic fibrosis.

A total of 258 respiratory tract specimens from patients with cystic fibrosis were inoculated onto nine different plated media, and the rates of recovery of potential pathogens were compared. Media included sheep blood agar, enriched chocolate agar, MacConkey agar for gram-negative bacilli, chocolate agar containing bacitracin for Haemophilus spp., bromcresol green agar for yeasts, cetrimide agar for Pseudomonas spp., sheep blood agar containing colistin and nalidixic acid for gram-positive cocci, mannitol salt agar for Staphylococcus aureus, and oxidation-fermentation agar containing 300 U of polymyxin B per ml and 2 U of bacitracin per ml (OF-PBL medium) for Pseudomonas cepacia. With two exceptions, all of these media proved useful in recovering potential pathogens from respiratory tract specimens from patients with cystic fibrosis. The two exceptions were cetrimide agar and colistin-nalidixic acid-supplemented sheep blood agar, which were found to be superfluous. In addition, the results of this study further delineated the prevalence of selected bacteria and fungi in respiratory tract secretions from patients with cystic fibrosis. In rank order of frequency of isolation, we recovered isolates of Pseudomonas aeruginosa, Haemophilus parainfluenzae, Candida albicans, S. aureus, Haemophilus influenzae, molds, members of the family Enterobacteriaceae, yeasts other than Candida albicans, miscellaneous gram-negative bacilli, beta-hemolytic streptococci, P. cepacia, and Streptococcus pneumoniae.     

Four-day incubation period for blood culture bottles processed with the Difco ESP blood culture system.

Blood culture records from 1994 to 1995 from five U.S. medical centers all using the Difco ESP continuous monitoring blood culture system were reviewed retrospectively. Among a total of 7,362 isolates of bacteria and yeasts, only 0.1% of possibly significant isolates would have been missed had blood cultures been routinely incubated for 4 days instead of the 5 days recommended by the manufacturer. Conversely, numerous contaminants, detected only on day 5, would have been eliminated by a 4-day incubation period.     

Sensititre autoreader for same-day breakpoint broth microdilution susceptibility testing of members of the family Enterobacteriaceae.

The Sensititre Autoreader system is an instrument-assisted broth microdilution susceptibility test procedure based on the detection of fluorogenic growth substrate metabolism by test bacteria with different concentrations of antimicrobial agents. In the current investigation, this system was assessed as a means for predicting the in vitro activity of 17 antimicrobial agents versus numerous species of the family Enterobacteriaceae and Pseudomonas aeruginosa by using a breakpoint broth microdilution test format. Same-day and overnight determinations of susceptibility were made with the Sensititre Autoreader system, and in both cases, the results were compared with those obtained with a manual overnight breakpoint broth microdilution susceptibility test. Among a total of 6,086 organism-antimicrobial agent comparisons with Enterobacteriaceae, concordance was noted between the results of the same-day Autoreader system and the manual overnight test in 94.4% of cases. The same-day Autoreader results with members of the Enterobacteriaceae other than Proteus spp. were determined after 4 h of incubation; with Proteus spp. the same-day Autoreader results were determined after 5 h of incubation. When the Enterobacteriaceae Autoreader results were determined after 18 h of incubation, concordance was noted in 97.2% of comparisons. Among a total of 1,377 organism-antimicrobial agent comparisons with P. aeruginosa after 18 h of incubation, agreement of results from the manual overnight test and the Autoreader system was achieved in 92.2% of cases.     

International Surveillance of Bloodstream Infections Due to Candida Species: Frequency of Occurrence and Antifungal Susceptibilities of Isolates Collected in 1997 in the United States, Canada, and South America for the SENTRY Program

An international program of surveillance of bloodstream infections (BSIs) in the United States, Canada, and South America between January and December 1997 detected 306 episodes of candidemia in 34 medical centers (22 in the United States, 6 in Canada, and 6 in South America). Eighty percent of the BSIs were nosocomial and 50% occurred in patients hospitalized in an intensive care unit. Overall, 53.3% of the BSIs were due to Candida albicans, 15.7% were due to C. parapsilosis, 15.0% were due to C. glabrata, 7.8% were due to C. tropicalis, 2.0% were due to C. krusei, 0.7% were due to C. guilliermondii, and 5.8% were due to Candida spp. However, the distribution of species varied markedly by country. In the United States, 43.8% of BSIs were due to non-C. albicans species. C. glabrata was the most common non-C. albicans species in the United States. The proportion of non-C. albicans BSIs was slightly higher in Canada (47.5%), where C. parapsilosis, not C. glabrata, was the most common non-C. albicans species. C. albicans accounted for 40.5% of all BSIs in South America, followed by C. parapsilosis (38.1%) and C. tropicalis (11.9%). Only one BSI due to C. glabrata was observed in South American hospitals. Among the different species of Candida, resistance to fluconazole (MIC, ≥64 μg/ml) and itraconazole (MIC, ≥1.0 μg/ml) was observed with C. glabrata and C. krusei and was observed more rarely among other species. Isolates of C. albicans, C. parapsilosis, C. tropicalis, and C. guilliermondii were all highly susceptible to both fluconazole (99.4 to 100% susceptibility) and itraconazole (95.8 to 100% susceptibility). In contrast, 8.7% of C. glabrata isolates (MIC at which 90% of isolates are inhibited [MIC₉₀], 32 μg/ml) and 100% of C. krusei isolates were resistant to fluconazole, and 36.9% of C. glabrata isolates (MIC₉₀, 2.0 μg/ml) and 66.6% of C. krusei isolates were resistant to itraconazole. Within each species there were no geographic differences in susceptibility to fluconazole or itraconazole.     

In vitro chloramphenicol susceptibility testing of Haemophilus influenzae: disk diffusion procedures and assays for chloramphenicol acetyltransferase.

The activity of chloramphenicol against 100 different strains of Haemophilus influenzae was assessed by a macrotube broth dilution technique and by a standardized disk diffusion method using both enriched chocolate agar (CHOC) and Mueller-Hinton agar containing 1.0% hemoglobin and 1.0% IsoVitaleX (BBL Microbiology Systems, Cockeysville, Md.) supplement (CHOC-MHA). Filter disks containing 30 micrograms of chloramphenicol were used with the disk diffusion procedure. The following zone diameter interpretive criteria were defined: CHOC-MHA, less than or equal to 25 mm = resistant [corrected] and greater than or equal to 26 mm = susceptible [corrected]; CHOC, less than or equal to 28 mm = resistant [corrected] and greater than or equal to 29 mm = susceptible [corrected]. all of the H. influenzae strains examined were also characterized by using two rapid assays for chloramphenicol acetyltransferase (CAT) activity: a 1-h tube method (t-CAT) and a 30-min procedure which used commercially available reagent-impregnated disks (d-CAT). The t-CAT procedure was found to be significantly more accurate than the d-CAT procedure as a means for demonstrating production of CAT.