Richard B. Clark

Virulence ofAeromonas species as assessed through mouse lethality studies

The relative virulence of 32Aeromonas isolates, primarily of clinical origin, were evaluated for mouse lethality by intraperitoneal inoculation of 107 CFU into albino mice. Three categories could be distinguished on the basis of this assay, including a highly virulent group (80%–100% mortality), a low to moderate virulence category (20%–60% mortality), and strains that were completely avirulent. Of theA. sobria isolates tested, 82% fell into the highly virulent category (P<0.005), whereasA. hydrophila strains were intermediate in virulence potential, andA. caviae strains studied were avirulent. There was no apparent correlation between highly virulentAeromonas isolates and phenotypes associated with enterotoxigenicity, hemolytic activity, cytotoxin production, or serum resistance; this suggests that a cell surface property may be important in mouse pathogenicity. The results of these studies indicate that mouse lethality assays may be an appropriate model for the study of invasive disease clinically produced byA. sobria andA. hydrophila.

Comparison of susceptibility test methods to detect penicillin-resistant Streptococcus pneumoniae

The detection of penicillin-resistant Streptococcus pneumoniae was assessed by six different methods: agar dilution, oxacillin screen by disk diffusion, E-test, and three overnight microdilution test methods that included commercial panels from MicroScan and Micro Media and in-house-made conventional panels using a commercial Haemophilus test medium (HTM) broth. Of the 52 pneumococcal isolates tested, 12 were resistant, 16 were relatively resistant, and 24 were susceptible to penicillin as defined by the reference agar dilution method. The oxacillin screen detected as resistant all 28 resistant and relatively resistant strains. The percentage of penicillin-resistant isolates detected by each minimum inhibitory concentration (MIC) test method was as follows: E-test (100%), Micro Media (75%), MicroScan (0%), and HTM (0%). With the relatively resistant isolates, the detection percentage was as follows: E-test (88%), Micro Media (94%), MicroScan (69%), and HTM (69%). In conclusion, the E-test and Micro Media MIC tests are acceptable confirmatory tests for detecting penicillin resistance among S. pneumoniae isolates.

In vitro susceptibilities of Edwardsiella tarda to 22 antibiotics and antibiotic-β-lactamase-inhibitor agents

The in vitro susceptibilities of 22 isolates of Edwardsiella tarda were studied with 22 antibiotics and antibiotic-beta-lactamase-inhibitor agents. Results indicated that all isolates were susceptible to the aminoglycosides, cephalosporins, penicillins, imipenem, aztreonam, ciprofloxacin, and antibiotic-beta-lactamase-inhibitor agents. Each strain produced a beta-lactamase even though no resistance was detected to the beta-lactams.

Comparison of five different susceptibility test methods for detecting antimicrobial agent resistance among Haemophilus influenzae isolates

The detection of antimicrobial agent resistance among ninety-eight Haemophilus influenzae isolates was assessed by six different antibiotic test methods: agar dilution on Mueller-Hinton agar supplemented with 5% lysed horse blood (MH-LHB), E-test using both Haemophilus test medium (HTM) agar and chocolate Mueller-Hinton (CMH) agar plates, Vitek Haemophilus susceptibility cards, and three overnight microdilution systems that included two commercial systems, Micro-Media and MicroScan, and the reference broth microdilution method using HTM broth. Agents tested in the study included ampicillin, amoxicillin/clavulanic acid (A/C), cefaclor, cefuroxime, cefotaxime, ceftriaxone, chloramphenicol, and trimethoprim/sulfamethoxazole. Both the reference HTM microbroth dilution method and agar dilution correctly classified all nine of the beta-lactamase negative ampicillin resistant (BLNAR) isolates. Each of the other test methods failed to detect one of the BLNAR strains, either because of growth failure (Micro-Media and MicroScan) or miscategorization of an isolate as susceptible (E-Test HTM, E-Test CMH, and Vitek). None of the test methods detected all six isolates identified as A/C resistant by HTM microbroth dilution. Of the remaining antimicrobials tested, ampicillin and cefuroxime yielded data that could be compared by all test methods. The very major, major, and minor errors for these two antimicrobials in comparison to the reference HTM microdilution method were as follows: Micro-Media (1.7%, 0%, and 4.8%); MicroScan (11.9%, 0%, and 8.1%); E-Test HTM (1.6%, 0%, and 2.0%); E-Test CMH (1.6%, 1.6%, and 4.6%); Vitek (8.1%, 0%, and 3.1%); and agar dilution on MH-LHB (0%, 0%, and 4.6%). Micro-Media and MicroScan panels failed to support the growth of 4.1% and 5.1% of the isolates, respectively.

Attachment of Mesophilic Aeromonads to Cultured Mammalian Cells

The interaction between mesophilic aeromonads and cultured mouse adrenal cells was examined. Preliminary experiments indicated that aeromonad attachment was dependent upon inoculum size, incubation time, and incubation temperature. Optimal attachment was observed after 30 min of incubation at 37°C with an inoculum size of 1×107 CFU. Heat-killed and formalin-treated organisms did not attach to the cultured cell system. The attachment of aeromonads to the mammalian cell surface was confirmed by light and scanning electron microscopy. Aeromonad attachment correlated both with the presence of pili and the specific aeromonad species, but not with hydrophobicity or the ability to autoagglutinate. Piliated strains were more likely to show high or moderate attachment.Aeromonas sobria, A. hydrophila, andA. veronii showed a greater ability to bind adrenal cells than didA. caviae. Removal of the pili from twoA. sobria isolates markedly reduced their attachment. In contrast, oneA. hydrophila isolate was strongly adherent after the removal of pili. The hemagglutination patterns produced byA. sobria and the other aeromonad species were distinctly different, but potentially predictive of the ability of aeromonads to attach to cultured mouse adrenal cells. These studies indicate that multiple mechanisms are important for the attachment of mesophilic aeromonads to mammalian cells. This model may prove useful for studying the pathogenesis of aeromonad infections.

Increased susceptibility of gentamincin-resistantPseudomonas aeruginosa to human sera

A total of 40 clinical strains ofPseudomonas aeruginosa were tested for in vitro resistance to the bactericidal action of pooled normal human sera (PHS). Getamicin-resistantP. aeruginosa (GRPA) were found to be significantly more susceptible to the killing action of 6% PHS than their gentamincin-sensitiveP. aeruginosa (GSPA) counterparts. In vitro mutants of GRPA were also more susceptible to PHS than their progenitor GSPA strain. The increased susceptibility of GRPA to PHS may help explain their lack of dissemination to internal body sites.

In Vitro Sensitivity of Candida (Torulopsis) glabrata to Clotrimazole

ABSTRACT: Vulvovaginitis caused by Candida (Torulopsis) glabrata is often refractory to intravaginal imidazole therapy. Clotrimazole achieves its fungistatic activity for Candida albicans and C. glabrata by inhibiting different steps in intermediary cell metabolism. For C. glabrata, alkylation precedes dimethylation. The possibility that this altered sequence might account for the relative therapeutic nonresponsiveness was studied by determining comparative minimal inhibitory concentrations (MICs) of clotrimazole. In vitro analyses of ten strains of C. glabrata and 30 control strains of C. albicans performed using both agar and broth dilution tests revealed that fourfold lower MICs were consistently demonstrable with C. glabrata, irrespective of inoculum size. The data suggest that clinical difficulties encountered in the therapy of torulopsis vulvovaginitis probably represent the inability of intravaginal medication to eradicate urethral/urinary bladder colonization and subsequent reinfection rather than true therapeutic failures.

Isolation of Serratia plymuthica from a Human Burn Site

The saprophytic bacterium Serratia plymuthica was recovered from a facial wound (burn) site of a pediatric patient. The clinical significance of the organism was undetermined due to its apparent eradication from this location by therapy with topical 1% silver sulfadiazine. Seeding of the burn with S. plymuthica may have occurred from contaminated moisture sometimes found on and around steam radiators.