Pseudomonas aeruginosa NDM-1 (CDC & FDA AR Isolate Bank #0246), Lung Infection Model, CFU Data
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OVERVIEW
DOCUMENTS/REFERENCES
BIOLOGICAL INFORMATION
BACKGROUND INFORMATION
This model assesses the antimicrobial efficacy of test articles in a lung infection model with carbapenem-resistant Pseudomonas aeruginosa. Microbial counts are measured. This carbapenem-resistant strain produces NDM-1 carbapenemase. It is resistant to β-lactam antibiotics, aminoglycoside, and quinolone. It is susceptible to colistin.
SPECIES
Pseudomonas aeruginosa
STRAIN COMMON NAME
AR Bank #0246
ORGANISM
Gram-negative Bacteria
INCUBATION CONDITIONS
Aerobic
NOTABLE ANTIBIOTIC RESISTANCE GENE
NDM-1
GENOMIC ACCESSION NUMBER
SAMN04901636
SOURCE
STRAIN SOURCE
CDC & FDA AR Isolate Bank
STRAIN SOURCE ID
AR Bank #0246
ASSAY INFORMATION
ASSAY TYPE
In Vivo
TESTING INFORMATION
PROCEDURE SUMMARY
Groups of 5 female specific-pathogen-free ICR (CD-1) mice weighing 22 ± 2 g are used. Animals are immunosuppressed by two intraperitoneal injections of cyclophosphamide. The first injection (150 mg/kg IP) is at 4 days before infection (day –4) and the second injection (100 mg/kg IP) is at 1 day before infection (day –1). On day 0, mice are anesthetized with pentobarbital (50 mg/kg IV) and are inoculated intranasally (0.05 mL/lung) into the lung with 1.0 x 10⁶ CFU/mouse of exponentially growing Pseudomonas aeruginosa (AR Bank #0246) culture. Vehicle and/or test substances are then administered at 2 and 14 hours later per the specification of the study design table. At 26 hours (18 - 20 hours for vehicle) after inoculation, animals are humanely euthanized with CO₂ asphyxiation and the lung tissues are harvested from each test animal. The removed lung tissues are homogenized in 1 mL of PBS, pH 7.4, with a polytron homogenizer. Homogenates (0.1 mL) are used for 10-fold serial dilutions and plated onto nutrient agar for colony count determination.
Note. This infection is lethal within the 26-hour time period if insufficiently treated. Mice of the vehicle group will be euthanized at 18 - 20 hours after infection because they will otherwise succumb to the infection. Mice of treatment groups with insufficient antimicrobial activity may succumb to the infection prior to the 26-h scheduled sacrifice time. Bacterial counts in tissues will not be determined in animals found dead prior
CRITERIA FOR SIGNIFICANCE
A significant (p < 0.05) decrease in the bacterial counts of the treated animals compared to the vehicle control group or the baseline control group was considered a significant difference.
TURNAROUND TIME
STANDARD
20 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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