Neisseria gonorrhoeae Azithromycin Non-Susceptible (CDC & FDA AR Isolate Bank #0181-StrR), Vaginal Infection Model, CFU Data
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BIOLOGICAL INFORMATION
BACKGROUND INFORMATION
This model assesses the efficacy of test articles for protection against a vaginal infection with azithromycin non-susceptible N. gonorrhoeae clinical isolate. It may be used for evaluating small molecules, vaccines and biologics.
SPECIES
Neisseria gonorrhoeae
STRAIN COMMON NAME
AR-BANK#0181-StrR
ORGANISM
Gram-negative Bacteria
INCUBATION CONDITIONS
Aerobic
SOURCE
STRAIN SOURCE
CDC & FDA AR Isolate Bank
STRAIN SOURCE ID
AR-BANK#0181
ASSAY INFORMATION
ASSAY TYPE
In Vivo
TESTING INFORMATION
PROCEDURE SUMMARY
Groups of 5 ovariectomized BALB/c mice 5 weeks of age are used. Animals are subcutaneously injected with a water soluble form of estradiol at 0.23 mg/mouse on day -2, 0, 2, 4 and 6. Starting from day -2 and following inoculation to the study end, mice are dosed twice daily with streptomycin (1.2 mg/mouse) and vancomycin (0.6 mg/mouse) by IP injection to minimize the vaginal flora. Trimethoprim sulfate 0.04 g/100 mL is administered in drinking water. At day 0, mice are anesthetized by pentobarbital (50 mg/kg) through IP injection, and then inoculated intravaginally with N. gonorrhoeae (AR-BANK#0181-17). Before inoculation, the mouse vagina is first rinsed with 30 µL of 50 mM HEPES (pH 7.4) followed by inoculation with 20 µL gonococci suspension in PBS containing 0.5 mM CaCl₂ and 1 mM MgCl₂. Test substance (IP, PO, IV, or SC upon request) and/or vehicle (1% Tween 80 or 0.9% NaCl, 10 mL/kg) are administered two hours after inoculation. Mice are euthanized with CO₂ asphyxiation at time points (2 h and 1 day are standard; 3 or 7 days upon request). Lavage is performed with 400 µL GC broth containing 0.05% saponin to recover vaginal bacteria. The bacterial suspensions are plated onto chocolate agar to determine the N. gonorrhoeae counts.
CRITERIA FOR SIGNIFICANCE
An agent is found to be bactericidal at a given concentration if treatment results in a 2-log reduction in counts over 26 hr relative to the baseline inoculum.
TURNAROUND TIME
STANDARD
30 Days
CLINICAL RELEVANCE
THERAPEUTIC AREA
Infectious Disease
ADDITIONAL INFORMATION
BRAND
PDS
TESTING LOCATION
Taiwan - Taipei
OTHER INFORMATION
We will readily accommodate client-specified alterations. These studies are performed at our AAALAC accredited BSL-2 laboratory in Taipei, Taiwan. All aspects of this work are performed in general accordance with the Guide for the Care and Use of laboratory animals (National Academy Press, Washington, DC, 2011). The study protocol was approved by the Pharmacology Discovery Services IACUC and is performed with the oversight of veterinarians to assure the humane treatment of laboratory animals.
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