Development of a Selective-Differential Nutrient Medium for the Isolation of Bordetella bronchiseptica
DOI:
https://doi.org/10.15407/Keywords:
Bordetella bronchiseptica, selective medium, differential medium, bacteriological diagnosisAbstract
The diagnosis of infectious diseases is crucial for monitoring animal welfare and ensuring sustainable epizootic health in different regions. This study aimed to experimentally validate the development of a selective-differential nutrient medium for isolating Bordetella bronchiseptica and to assess its effectiveness. Methods. The Bordetella bronchiseptica ATCC 4617 strain was used in the study. The temperature, pH, nutrient base, selective components, and differential markers were optimized stepwise. The test was performed on 69 clinical samples (47 PCR-positive and 22 PCR-negative). We conducted a comparison of isolation frequencies using Fisher's exact test and McNemar's test. Results. The optimal cultivation parameters for Bordetella bronchiseptica were determined to be 36.0 ± 0.5 °C and pH 7.1 ± 0.1. A nutrient medium was developed based on the results of the stepwise optimization of the nutrient base and selective and differential components (g/L of distilled water): enzymatic peptone – 20.0; yeast extract – 5.0; casein – 5.0; urea – 5.0; 2,3,5-triphenyltetrazolium chloride (TTC) – 0.01; cephalexin – 0.003; bromothymol blue – 0.02; agar – 15.0. The developed medium provided an isolation frequency of 72.3%, exceeding Regan-Lowe (61.7%) and charcoal-blood agar (CBA) (51.1%). A statistically significant advantage over CBA was found (p < 0.05). Conclusions. The proposed medium increases the effectiveness of culture-based diagnostics and enables the initial phenotypic evaluation of isolates during primary cultivation.
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