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> Home > Lyme kruidentherapie versus antibiotica

Lyme kruidentherapie versus antibiotica

To clarify the impact of the Lyme Plus Protocol (APP Plus, DTC Plus and TBB Plus) on Borrelia burgdorferi that has been cultured in vitro at +37°C

Hypothesis

An efficient therapy, antimicrobial agents that can eliminate all the forms of B. burgdorferi have to be found.
Efflux pumps give Borrelia resistance to several antibiotics by antimicrobial efflux, and combining antimicrobial therapy with non-toxic compounds that inhibit efflux could increase antimicrobial efficiency against all the pleomorphic forms of B. burgdorferi.

Study objectives

Studying the effects of antibiotics and herbal compounds of the Lyme Plus Protocol on the different pleomorphic forms of B. burgdorferi. The aim of these experiments is to use sensitivity and specificity studies to find out what kind of effects the compounds individually have on different pleomorphic forms of B. burgdorferi. MIC and MBC of the individual compounds will be assessed.

Initial Experimental Setup


1. Minimum Inhibitory Concentration (MIC)

MIC will be determined according to Ates et al. (2010) with modifications to evaluate the antimicrobial properties of the compounds. The lowest antibiotic concentration at which Borrelia fails to grow will be interpreted as the MIC. 1 x 10e6 cells in volume of 200 ul. Cultured in vitro at +37°C for 24, 48, 72, 96 hrs post-exposure to antimicrobial agent. Fluorescent reading from the bacteria is recorded.
Pleomorphic forms are observed with the determine MIC.
Concentrations of antibiotics and Lyme Plus Protocol compounds were 5, 50, 100, 200, 400, 800 and 100 µg/ml or otherwise stated
2. Minimum Bactericidal Concentration (MBC)

MBC will be determined according to Ates et al. (2010) with modifications. The cultures with antimicrobials that do not have bacterial growth after 72 h will be subcultured for 3 weeks in vitro at +37°C in fresh media.
The lowest antimicrobial concentration that will not contain live bacteria after this subculturing step will be considered as the MBC.
Results: MIC of antibiotics


MIC 100 μg/ml confirmed by MBD
No Pleomorphic forms but Damaged Cells

Relative fluorescence units (RFU) of fluorescent B. burgdorferi strain with azithromycin, clindamycin and doxycycline over time when background media subtracted. B. burgdorferi spirochetes (1 x 106/ml) were various concentrations of the antibiotics. H2O2 was used as a control for death. Representative image of the similar results obtained from all three antibiotics. Data is presented from three different experiments.

Results: MICs of Lyme Plus Protocol Compounds


MIC 5 μg/ml confirmed by MBD
No Pleomorphic forms but Damaged Cells

Relative fluorescence units (RFU) of fluorescent B. burgdorferi strain APP Plus, DTC Plus and TBB Plus over time when background media subtracted. B. burgdorferi spirochetes (1 x 106/ml) were various concentrations of the antibiotics. H2O2 was used as a control for death. Representative image of the similar results obtained from all three antibiotics. Data is presented from three different experiments.
 

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