Uptake of and resistance to the antibiotic berberine by individual dormant, germinating and outgrowing bacillus spores as monitored by laser tweezers raman spectroscopy

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

Shiwei,Yu,Jing,Suvira,Milomir,Setlow,Peter,Li,Yong Wang (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

Abstract: Berberine, an alkaloid originally extracted from the plant Coptis chinensis and other herbplants, has been used as a pharmacological substance for many years. The therapeuticeffect of berberine has been attributed to its interaction with nucleic acids and blocking celldivision. However, levels of berberine entering individual microbial cells minimal for growthinhibition and its effects on bacterial spores have not been determined. In this work thekinetics and levels of berberine accumulation by individual dormant and germinated sporeswere measured by laser tweezers Raman spectroscopy and differential interference andfluorescence microscopy, and effects of berberine on spore germination and outgrowth andspore and growing cell viability were determined. The major conclusions from this work arethat: (1) colony formation from B. subtilis spores was blocked ~ 99% by 25 μg/mL berberineplus 20 μg/mL INF55 (a multidrug resistance pump inhibitor); (2) 200 μg/mL berberine hadno effect on B. subtilis spore germination with L-valine, but spore outgrowth was completelyblocked; (3) berberine levels accumulated in single spores germinating with 25 μg/mLberberine were > 10 mg/mL; (4) fluorescence microscopy showed that germinated sporesaccumulated high-levels of berberine primarily in the spore core, while dormant sporesaccumulated very low berberine levels primarily in spore coats; and (5) during germination,uptake of berberine began at the time of commitment (T1) and reached a maximum after thecompletion of CaDPA release (Trelease) and spore cortex lysis (Tlysis).

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Other

Language: English

Date: 2015

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