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A new selective enrichment procedure for isolating Pasteurella multocida from avian and environmental samples
M K Moore1, L Cicnjak-Chubbs, R J Gates
1National Biological Survey, National Wildlife Health Research Center, Madison, Wisconsin 53711.
Abstract:
A selective enrichment procedure, using two new selective media, was developed to isolate Pasteurella multocida from wild birds and environmental samples. These media were developed by testing 15 selective agents with six isolates of P. multocida from wild avian origin and seven other bacteria representing genera frequently found in environmental and avian samples. The resulting media--Pasteurella multocida selective enrichment broth and Pasteurella multocida selective agar--consisted of a blood agar medium at pH 10 containing gentamicin, potassium tellurite, and amphotericin B. Media were tested to determine: 1) selectivity when attempting isolation from pond water and avian carcasses, 2) sensitivity for detection of low numbers of P. multocida from pure and mixed cultures, 3) host range specificity of the media, and 4) performance compared with standard blood agar. With the new selective enrichment procedure, P. multocida was isolated from inoculated (60 organisms/ml) pond water 84% of the time, whereas when standard blood agar was used, the recovery rate was 0%.
Insights
A new selective enrichment method effectively isolates Pasteurella multocida from wild birds and environmental samples. This advancement significantly improves detection rates compared to standard methods.
Area of Science:
- Veterinary Microbiology
- Wildlife Disease Ecology
- Bacteriology
Background:
- Pasteurella multocida is a significant pathogen in avian species, impacting wild bird populations.
- Accurate isolation of P. multocida from environmental and avian samples is crucial for disease surveillance and control.
- Existing isolation methods may lack the sensitivity and specificity required for diverse sample types.
Purpose of the Study:
- To develop and evaluate novel selective enrichment media for the isolation of Pasteurella multocida.
- To enhance the detection of P. multocida from challenging matrices like wild bird carcasses and environmental water samples.
- To compare the performance of the new media against conventional techniques.
Main Methods:
- Development of two selective media: Pasteurella multocida selective enrichment broth and Pasteurella multocida selective agar.
- Testing of 15 selective agents against P. multocida and common environmental/avian bacteria.
- Evaluation of media selectivity, sensitivity, host range specificity, and performance in isolating P. multocida from inoculated pond water and avian carcasses.
- Comparison with standard blood agar for P. multocida recovery.
Main Results:
- The new selective media demonstrated high selectivity and sensitivity for P. multocida.
- P. multocida was recovered from 84% of inoculated pond water samples using the new procedure.
- Standard blood agar yielded a 0% recovery rate for P. multocida from the same inoculated pond water samples.
- The developed media significantly outperformed standard blood agar in detecting low bacterial loads.
Conclusions:
- The novel selective enrichment procedure offers a superior method for isolating Pasteurella multocida from wild birds and environmental sources.
- This technique provides a valuable tool for wildlife disease monitoring and epidemiological studies.
- The enhanced sensitivity and specificity of the new media address limitations of traditional isolation methods.
