Lyme disease or Lyme borreliosis is a tickborne multisystemic
disease caused by different species of Borrelia.
antibiotics is the primary treatment for this disease, however,
relapse often occurs when antibiotic treatment is discontinued.
One possible reasoning is that Borrelia becomes resistant to
antibiotic treatment by converting from their spirochete form to
several defensive forms such as cysts and biofilms.
studies on antibiotic resistant pathogens demonstrated that certain herbal extracts could have significant antimicrobial,
antiprotozoal, antiviral, properties and in addition to that they are proven to be nontoxic and can be taken safely for longer period of time.
3 The aim is to test the effectiveness of three different
medicinal agents (Cumanda, Lakato and Stevia) on the different
morphological forms of Borrelia burgdorferi using fluorescent,
dark field microscopic techniques, special staining techniques
including BacLight viability staining methods and by total
carbohydrate analysis. As antibiotics are proven to be ineffective
in eliminating all the morphological forms, natural antimicrobial
agents might provide a more effective therapeutic option for
Lyme disease patients.
We are grateful to the University of New Haven, Nutramedix LLC
(Jupiter, Florida), TBDA (Turn the Corner Foundation), LDA, Lyme
Research Alliance, Lyme disease. org (CALDA) and Schwartz
Foundation for all their support.
In summary, the treatment involving Stevia, resulted in a significant reduction in the numbers of spirochetal form by 60% and the cyst forms by 98%. Doxycycline reduced the spirochetal forms by 77% but showed a marked increase in the cyst form at higher concentration. The treatment with Lakato also show similar trends but less effective than Stevia. Furthermore, Cumanda at a higher
dilution (1:900 dilution) reduced the spirochete form by 24% and the cyst forms by 5%.When quantitative effects on biofilm were evaluated, Cumanda reduced the formation of colonies by 43%
Priyanka A.S Theophilus M.S.
, Divya Burugu B.S.
, Akhila Poururi B.S.
, David F. Luecke B.S.
, Eva Sapi Ph.D1
Lyme Disease Research Group, Department of Biology and Environmental Sciences,
University of New Haven, CT, 06516
Figure1: The in vitro susceptibility of spirochete and cyst forms of B31 Borrelia burgdorferi to Cumanda, Lakato and Stevia and to doxycycline (25 µg/ml) for 96 hours'
treatment period using direct cell counting and dark field morphological evaluation methods. P- values <0.05 indicates statistical significance (*).
The three morphological forms of Borrelia burgdorferi B31
(spirochetes, round bodies and biofilm) were analyzed in this
1 The antimicrobial sensitivity of spirochetal and cyst forms
was tested using dark field microscopy (direct cell counting).
Qualitative effects of antimicrobials against biofilms were assessed
using fluorescent microscopy and quantitative analysis on biofilm
was performed following the Total Carbohydrate Assay using
standard published technique.
4 The three antimicrobial agents
tested in this study has shown significant antimicrobial effects in
Therefore in conclusion the treatment involving Stevia show a
significant reduction in both the spirochetal and cyst forms, while
Cumanda had the most significant effect in reducing the colony
formation. Moreover, our results demonstrated that herbal agents
had significant effects on all forms of Borrelia burgdorferi, implying
that these antimicrobial agents could provide an effective therapeutic
approach for Lyme disease patients.
1) Sapi, E., Kaur, N., Anyanwu, S., Luecke, D.F., Datar, A., Patel, S., Rossi, M., and Stricker,
R.B. “Evaluation of In-Vitro Antibiotic Susceptibility of Different Morphological
Forms of Borrelia burgdorferi” (2011), Infection and Drug Resistance, 4, 97-113.
2) Sapi, E., Bastian, S. L., Mpoy, C.M., Scott,S., Rattelle, A., Pabbati, N., Poruri, A.,
Burugu,D., Theophilus, P. A. S., Pham, T. V., Datar, A., Dhaliwal, N. K., MacDonald, A.,
Rossi,M. J., Sinha, S. K., and Luecke, D. F. “Characterization of Biofilm Formation by
Borrelia burgdorferi In Vitro” (2012), PLoS ONE, 7 (10), 1-11.
3) Datar, A., Kaur, N., Patel, S., Luecke, D.F., and Sapi, E. “In Vitro Effectiveness of
Samento and Banderol Herbal Extracts on the Different Morphological Forms of
Borrelia burgdorferi” (2010), Townsend Letter.
4) Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., and Smith, F. “Colorimetric
method for determination of sugars and related substances” (1956), Analytical
Chemistry, 28, 350-356.
5) Kvist, L. P., Christensen, S. B., Rasmussen, H. B., Mejia, K., and Gonzalez, A.
“Identification and evaluation of Peruvian plants used to treat malaria and
leishmaniasis” (2006), Journal of Ethnopharmacology, 106 (3), 390-402.
6) Cohen, H.A., Varsano, I., Kahan, E., Sarrell, E.M., and Uziel, Y. “Effectiveness of an
herbal Preparation Containing Echinacea, Propolis, and Vitamin C in Preventing
Respiratory Tract Infections in Children” (2004), Archives of Pediatrics and Adolescent
Medicine, 158 (3), 217-221.
7) Ghosh, S., Subudhi, E., and Nayak, S. “Antimicrobial Assay of Stevia rebaudiana
Bertoni Leaf Extracts against 10 Pathogens” (2008),