Effect of short- and long-term antibiotic exposure on the viability of Mycobacterium avium subsp. paratuberculosis as measured by propidium monoazide F57 real time quantitative PCR and culture
Language English Country England, Great Britain Media print-electronic
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
22704136
DOI
10.1016/j.tvjl.2012.05.002
PII: S1090-0233(12)00190-6
Knihovny.cz E-resources
- MeSH
- Amphotericin B pharmacology MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Azides metabolism MeSH
- Time Factors MeSH
- Real-Time Polymerase Chain Reaction methods MeSH
- Nalidixic Acid pharmacology MeSH
- Mycobacterium avium subsp. paratuberculosis drug effects growth & development isolation & purification metabolism MeSH
- Paratuberculosis diagnosis microbiology MeSH
- Colony Count, Microbial methods MeSH
- Ruminants microbiology MeSH
- Propidium analogs & derivatives metabolism MeSH
- Vancomycin pharmacology MeSH
- Dose-Response Relationship, Drug MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Amphotericin B MeSH
- Anti-Bacterial Agents MeSH
- Azides MeSH
- Nalidixic Acid MeSH
- propidium monoazide MeSH Browser
- Propidium MeSH
- Vancomycin MeSH
Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of paratuberculosis in ruminants, has a lipid-rich cell wall which facilitates its survival and persistence in the environment. This property of the organism is exploited when it is cultured as decontaminating agents and antibiotics are used to suppress the growth of contaminating microflora, but such treatments can also negatively affect the isolation of MAP itself. The objective of this study was to assess the effect of the 'VAN' antibiotics (vancomycin, amphotericin B and nalidixic acid) on the viability of MAP using a propidium monoazide real time quantitative PCR (PMA qPCR) and culture. Long-term (5 week) treatment with VAN antibiotics resulted in a larger decrease in bacterial numbers compared to short-term (3 day) exposure. The PMA qPCR assay indicated that 50 μg/mL of vancomycin, 50 μg/mL of nalidixic acid, and 200 μg/mL of amphotericin B were 'threshold' concentrations, respectively, above which the decline in the viability of MAP was statistically significant. Using culture, these threshold concentrations were 100 μg/mL of vancomycin, 50-100 μg/mL of nalidixic acid, and 100 μg/mL of amphotericin B, respectively. Given that the two methods were found to be comparable, the PMA qPCR is a potentially more convenient and effective alternative to culture in detecting MAP.
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