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In Vitro Effectiveness of Samento and Banderol Herbal Extracts on the Different Morphological Forms of Borrelia Burgdorferi

11/13/2017 Effectiveness of Samento and Banderol on Borrelia Burgdorferi (July 2010) Townsend Letter for Doctors & Patients 1/8
From the Townsend Letter
July 2010
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In Vitro Effectiveness of Samento and
Banderol Herbal Extracts on the Different
Morphological Forms of Borrelia
by Akshita Datar, Navroop Kaur, Seema Patel,
David F. Luecke, and Eva Sapi, PhD
Lyme Disease Research Group
University of New Haven
A tick-borne, multisystemic disease, Lyme borreliosis caused by the
spirochete Borrelia burgdorferi has grown into a major public health
problem during the last 10 years. The primary treatment for chronic
Lyme disease is administration of various antibiotics. However,
relapse often occurs when antibiotic treatment is discontinued. One
possible explanation for this is that B. burgdorferi become resistant to
antibiotic treatment, by converting from their vegetative spirochete
form into different round bodies and/or into biofilmlike colonies.
There is an urgent need to find novel therapeutic agents that can
eliminate all these different morphologies of B. burgdorferi. In this
study, two herbal extracts, Samento and Banderol, as well as
doxycycline (one of the primary antibiotics for Lyme disease
treatment) were tested for their in vitro effectiveness on several of the
different morphological forms of B. burgdorferi (spirochetes, round
bodies, and biofilmlike colonies) using fluorescent, darkfield
microscopic, and BacLight viability staining methods. Our results
demonstrated that both herbal agents, but not doxycycline, had very
significant effects on all forms of B. burgdorferi, especially when used
in combination, suggesting that herbal agents could provide an
effective therapeutic approach for Lyme disease patients.
Borrelia burgdorferi, the primary causative agent of Lyme disease, is a spirochetal
bacterium that can adopt different inactive forms, such as cystic and granular
forms (round bodies), as well as colonylike aggregates both in vivo and in vitro, in
the presence of unfavorable conditions such as exposure to the antibiotics
commonly used for treating Lyme borreliosis.1-4 Unfortunately, when B.
burgdorferi is in these inactive forms, conventional antibiotic therapy will not
destroy the bacteria.3 Still to date, the frontline treatment for Lyme disease is
administration of pharmaceutical antibiotics such as doxycycline, minocycline,
clarithromycin, penicillin G, and ceftriaxone.4,5 Many studies have shown that in
spite of continued and high-dose antibiotic therapy, chronic Lyme disease is not
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11/13/2017 Effectiveness of Samento and Banderol on Borrelia Burgdorferi (July 2010) Townsend Letter for Doctors & Patients 2/8
treated successfully in many cases.6 Also, in the absence of ongoing antibiotic
treatment, relapse is common.7,8 This means that even after antibiotic treatment,
the host immunity fails to prevent recurrence.8 One possible explanation for this
clinical observation is the presence of different morphological forms of B.
burgdorferi, which mayprotect it from the antibacterial therapy. Soon after
treatment, relapse is observed, most likely because the B. burgdorferi can revert to
the spirochetal form. Furthermore, the cost of antibiotic treatment, especially when
administered intravenously, is substantial. Antibiotic therapy may also cause
multiple undesirable side effects.9 Thus, there is an urgent need for novel, more
efficient, and more cost-effective treatment approaches that can efficiently
eliminate all forms of B. burgdorferi.
There is an alternative clinical treatment option gaining wide use, called Cowden
Condensed Support Program, that utilizes several herbal extracts designed to
eliminate microbes in Lyme disease patients. Richard Horowitz, MD, president of
the International Lyme and Associated Diseases Educational Foundation
(ILADEF), has prescribed this protocol for over 2000 of his patient and reports
that it has been effective for more than 70% of them. The two herbal agents from
the Cowden Condensed Support Program selected for this study are Samento (a
pentacyclic chemotype of Cat's Claw [Uncaria tomentosa] that does not contain
tetracyclic oxindole alkaloids), with reported antibacterial and antiviral properties,
and Banderol (Otoba sp.), known to have antibacterial, antiprotozoal and anti-
inflammatory effects.10-12 Both herbal agents are used during the first two months
of Cowden Condensed Support Program, then in rotation with other antimicrobials
for the duration of this 6-month protocol.
In this study, we evaluated these natural antimicrobial herbal extracts as well as
doxycycline (one of the primary pharmaceutical antibiotics for Lyme disease
treatment) for their potential effects on the different forms of B. burgdorferi.
The infectious B31strain of B. burgdorferi used in this study, obtained from
American Type Tissue Collection(ATCC# 35210), was culturedin 5% CO2 at 34
oC, in Barbour–Stoener–Kelly H (BSK H) medium supplemented with 6% rabbit
serum (Sigma, St. Louis, Missouri) to midlogarithmic stage (2 × 107 cells/ml).
Samento and Banderol were obtained from Nutramedix LLC (Jupiter, Florida).
Doxycycline was obtained from Sigma. A wide range of concentrations of
Samento and Banderol were initially tested to determine the effective
concentrations (1:100–1:1000 dilutions). For doxycycline, a concentration 10×
higher than the reported minimum bactericidal concentration (250 µg/ml) was
used.13 Triplicate test tubes containing BSK H medium, with and without the
11/13/2017 Effectiveness of Samento and Banderol on Borrelia Burgdorferi (July 2010) Townsend Letter for Doctors & Patients 3/8
appropriately diluted antimicrobial agents, were inoculated with a final density of
5 × 106 cells/ml of the test organism.
Direct cell counting methods with Petroff-Hausser counting chambers and
morphological studies using fluorescent and darkfield microscopic techniques, as
well as LIVE/DEAD BacLight Bacterial Viability Assay (Life Technologies Corp,
Carlsbad, California), were utilized to assess the effect of the antimicrobial agents.
For statistical analyses, one sample paired T-test was performed using NCSS
statistical software (NCSS LLC, Kaysville, Utah).
Samento & Banderol Herbal Extracts
Figure 1A
Figure 1B
11/13/2017 Effectiveness of Samento and Banderol on Borrelia Burgdorferi (July 2010) Townsend Letter for Doctors & Patients 4/8
Figure 1C
Figures 1: The in vitro susceptibility of the spirochete and round-body forms
of the B31 B. burgdorferi to Samento and Banderol extracts and to
doxycycline (250 µg/ml) for 96 hours' treatment period using direct cell
counting and darkfield morphological evaluation methods. (A) Samento
extract; (B) Banderol extract; (C) Samento + Banderol extracts. As a negative
control, 0.25% ethanol was a used. *P- values >0.05 indicates statistical
In the first set of experiments, we tested the in vitro susceptibility of the spirochete
and round-body forms of the B. burgdorferi B31 strain to Samento and Banderol
extracts for 96 hours, then direct cell counting and darkfield morphological
evaluation methods were used to measure the effects of the antimicrobial agents.
For both herbal extracts, the dilution of 1:400 most efficiently eliminated both the
spirochetal and round-body forms (Figure 1A and 1B). However, when we used
the combination of Samento and Banderol extracts, 1:300 dilution showed the
most effectiveness, and this concentration was chosen for further study (Figure
1C). As a negative control, 0.25% ethanol treatment was also included in all
experiments, because these herbal extracts contain ~25% ethanol to transport the
nutrients into the cells and for stability.
In these experiments, we also compared the effect of Samento and Banderol with
doxycycline, the most common antibiotic treatment agent for Lyme disease
treatment in a 96-hour treatment period. Our results showed that doxycycline (250
µg/ml) was very effective in eliminating the spirochetal form of B. burgdorferi, but
it significantly increased the round-body forms. Comparing this doxycycline data
with that of the herbal extracts, Banderol and the combination of Samento and
Banderol (1:300) were more efficient in eliminating both the spirochetal and
round-body forms of B. burgdorferi in vitro (Figures 1A–C).
In the next set of experiments, we evaluated the effect of the different
antimicrobial agents on biofilmlike colonies of B. burgdorferi. The cultures were
treated as described above for 96 hours and stained with BacLight fluorescent
viability stains, which can help visualize the effects of the antimicrobial agents on
11/13/2017 Effectiveness of Samento and Banderol on Borrelia Burgdorferi (July 2010) Townsend Letter for Doctors & Patients 5/8
the bacterial cells (Figure 2). The green fluorescent stain (SYTO 9, with
excitation/emission maxima of about 480/500 nm) colors healthy bacteria that
have intact membranes, thus staining live cells; and the red dye (propidium iodide
with excitation/emission maxima of about 490/635 nm) colors bacteria with
damaged membranes, by displacing the green dye, thus staining dead cells.
Figures 2: BacLight viability staining of B31 strain of B. burgdorferi after 96-
hour treatment using SYTO 9 green-fluorescent nucleic acid stain (live cells)
and propidium iodide, a red-fluorescent nucleic acid stain (dead cells). (A)
Control; (B) Samento (1:300 dilution); (C) Banderol (1:300 dilution); (D)
Samento + Banderol (1:300 dilution); (E) Doxycycline (250 µg/ml). All images
are taken at 40× magnification.
Figure 2A
Figure 2B
Figure 2C
In the absence of antimicrobial agents,
B. burgdorferi is forming biofilmlike
colonies (Figure 2A) with mainly live
bacterial cells. In the presence of
Samento extract (1:300), the colonies
were significantly smaller and less
organized (Figure 2B), but they did
stain with green dye, indicating that live
cells remained. In the presence of
Banderol extracts, the size of colonies
did not show any reduction; however,
the cells inside the colonies are >90%
In the presence of both herbal extracts,
no sign of any colony formation was
observed in the cultures, but we found
evidence of a few individual nonmotile
but green spirochetes and round bodies.
In the presence of doxycycline (250
µg/ml), the average colony size was
increased and contained mainly live
round-body forms.
In this study, our working hypothesis
was that for an efficient therapy, we
have to find antimicrobial agents that
can eliminate all the forms of B.
burgdorferi. During the course of
Borrelia infection, the bacteriumcan
shift among the different forms,
11/13/2017 Effectiveness of Samento and Banderol on Borrelia Burgdorferi (July 2010) Townsend Letter for Doctors & Patients 6/8
Figure 2D converting from the spirochete form to
the others when presented with an
unfavorable environment and reverting
to the spirochete when the condition is
again favorable for growth.1-4 To
successfully eradicate B. burgdorferi,
antimicrobial agents should eliminate
all those forms, including the
spirochetes, round bodies, and
biofilmlike colonies.
Here we have provided evidence that
two natural antimicrobial agents
(Samento and Banderol extracts) had
significant effect on all three known
forms of B. burgdorferi bacteria in vitro.
We have also demonstrated that
doxycycline, one of the primary
antibiotics used in the clinic to treat
Lyme disease, only had significant
effect on the spirochetal form of B.
Figure 2E
Our later results might provide some explanation for why relapse is so common
after discontinuing antibiotic therapy. For example, some of the recent reports on
animal experiments demonstrated that although pharma ceutical antibiotics are
effective in ameliorating disease, the infection may persist even after seemingly
effective therapy, which suggested that Borrelia may remain viable even after
antibiotic administration.14-15 If those pharmaceutical antibiotics only eliminate
one form of this bacterium, the other forms could be the source of the persistent
The other very important fact needs to be considered for an effective treatment for
Borrelia infection: this bacterium typically has a life span ranging from several
weeks to six to eight months; therefore, it may take six to eight months for even
one generation of Borrelia to become exposed to the antimicrobial for
elimination.16 Since the herbal extracts like Samento are reported to be nontoxic,
they can be safely taken daily for the long period of time necessary to thoroughly
eradicate Borrelia from an infected body.17
In summary, our study has provided in vitro research data on a novel treatment
11/13/2017 Effectiveness of Samento and Banderol on Borrelia Burgdorferi (July 2010) Townsend Letter for Doctors & Patients 7/8
approach using herbal antimicrobial agents to efficiently eradicate B. burgdorferi,
the Lyme disease bacterium.
Corresponding Author
Eva Sapi, PhD
University of New Haven
Department of Biology and Environmental Sciences
300 Boston Post Road
West Haven, Connecticut 06516
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cystic forms to motile spirochetes in vivo. Acta Pathol Microbiol Scand.
2. Brorson Ø, Brorson SH. In vitro conversion of Borrelia burgdorferi to cystic
forms in spinal fluid, and transformation to mobile spirochetes by incubation in
BSK-H medium. Infection. 1998;26:44–50.
3. Miklossy J, Kasas S, Zurn AD, McCall S, Yu S, McGeer PL. Persisting
atypical and cystic forms of Borrelia burgdorferi and local inflammation in Lyme
neuroborreliosis. J Neuroinflammation. 2008;25:5–40.
4. Brorson Ø, Brorson SH, Scythes J, MacAllister J, Wier A, Margulis L.
Destruction of spirochete Borrelia burgdorferi round-body propagules (RBs) by the
antibiotic tigecycline. Proc Natl Acad Sci U S A. 2009 Nov;106(44):18656–18661.
5. Burrascano J. Advanced topics in Lyme disease. In: Managing Lyme Disease.
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6. Krause PJ, Foley DT, Burke GS, Christianson D, Closter L, Spielman A.
Reinfection and relapse in early Lyme disease. Am Trop Med Hyg.
7. Klempner M, Linden MD, Hu T, J Evans J, et al. Two controlled trials of
antibiotic treatment in patients with persistent symptoms and a history of Lyme
disease. N Engl J Med. 2001;345:85–92.
8. Horowitz R. Classical and integrative medical approaches in chronic Lyme
disease: new paradigms in diagnosis & treatment. 8th Annual International Lyme
and Associated Diseases Society (ILADS) Conference; 2007 October.
9. Matsuura T, Shimizu Y, Fujimoto H, et al. Minocycline-related lupus.
10. Ccahuana-Vasquez RA, Santos SS, Koga-Ito CY, Jorge AO. Antimicrobial
activity of Uncaria tomentosa against oral human pathogens. Braz Oral Res. 2007
11. Valerio LG Jr, Gonzales GF. Toxicological aspects of the South American
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herbs cat's claw (Uncaria tomentosa) and Maca (Lepidium meyenii): a critical
synopsis. Toxicol Rev. 2005;24(1):11–35.
12. Weniger B, Robledo S, Arango GJ, et al. Antiprotozoal activities of Colombian
plants. J Ethnopharmacol. 2001 Dec;78(2–3):193–200.
13. Baradaran-Dilmaghani R, Stanek G. In vitro susceptibility of thirty Borrelia
strains from various sources against eight antimicrobial chemotherapeutics.
Infection. 1996 Jan–Feb;24(1):60–63.
14. Bockenstedt LK, Mao J, Hodzic E, et al. Detection of attenuated, noninfectious
spirochetes in Borrelia burgdorferi-infected mice after antibiotic treatment. J Infect
Dis. 2002; 186:1430–1437.
15. Barthold SW, Hodzic E, Imai DM, Feng S, Yang X, Luft BJ. Ineffectiveness of
tigecycline against persistent Borrelia burgdorferi. Antimicrob Agents Chemother.
2010 Feb;54(2):643–651.
16. Samuels DS and Radolf JD. Borrelia: Molecular Biology, Host Interaction and
Pathogenesis. Caister Academic Press; 2010.
17. Reinhard K-H. Uncaria tomentosa (Willd.) D.C.: Cat's claw, Una de Gato, or
Saventaro. J Alt Comp Med. 1999;5:143–151.
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June 9, 2010
... Samento (a TOA-free extract from Uncaria tomentosa) and Banderol (an extract from Otoba parvifolia) have been demonstrated to be effective against different morphological forms (spirochetes, round bodies, and biofilm-like colonies) of Borrelia burgdorferi in vitro [8], one of the Borrelia species causing Lyme borreliosis [9]. Whereas doxycycline, one of the most commonly used antibiotics to treat Lyme borreliosis, was only efficient against the spirochetal form, both extracts and especially the combination of them were also effective against the round body forms and biofilms, which are considered to be the source of the persistent Lyme disease [10,11]. ...
... Banderol is an extract from Otoba parvifolia bark containing 20-24% ethanol. The maximum concentration used for both extracts was set to a dilution of 1:100 due to several reasons: (1) this dilution contains 0.2% ethanol, which does not influence the assays conducted; (2) dilutions of 1:400 showed best activity against the round-body forms of Borrelia burgdorferi in vitro, whereas higher concentrations were only effective against the spirochete form [8]; (3) the manufacturer of Samento and Banderol recommends to take a maximum of 30 drops (=1.5 ml) in about 100 ml water, which represents a dilution of 1:150 (=0.7%). Thus, it seemed unreasonable to test higher concentrations than a dilution of 1:100 (=1%). ...
Full-text available
Samento (extract from Uncaria tomentosa) and Banderol (extract from Otoba parvifolia) have been demonstrated to have anti-inflammatory and antimicrobial properties, e.g., against different morphological forms of Borrelia burgdorferi. However, there is hardly any data on the pharmacological safety of these two herbal medicines. This in vitro study aimed at scrutinizing their possible characteristics as perpetrators in pharmacokinetic herbal–drug interactions. Inhibition of cytochrome P450 enzymes (CYPs) was quantified by commercial kits and inhibition of drug transporters by use of fluorescent probe substrates. Induction was quantified by real-time RT-PCR and activation of pregnane x receptor (PXR) and aryl hydrocarbon receptor (AhR) by reporter gene assays. Organic anion transporting polypeptide 1B1 (OATP1B1) (IC50 = 0.49 ± 0.28%) and OATP1B3 (IC50 = 0.65 ± 0.29%) were potently inhibited by Banderol, but only weakly by Samento. CYP3A4 was inhibited about 40% at a Samento concentration of 1%. Samento significantly induced mRNA expression of CYP2J2, UGT1A3, UGT1A9, ABCB1, and SLCO1B1 and strongly activated PXR, but hardly AhR. In conclusion, the perpetrator profiles of Samento and Banderol for herb–drug interactions completely differ. Clinical studies are strongly recommended to clarify whether the effects observed in vitro are of clinical relevance.
... Some antibiotics may be able to arrest growth and metabolic activity in free-living forms of a bacterium but may also trigger evasive tactics by the microbe to form biofilm and to enter a quiescent state as 'persister' cells. 50 Multifaceted natural treatments combined with immune support has shown promise, and herbal extracts, many used in traditional folk medicine, are showing promise in inhibiting biofilm formation. [51][52][53][54][55] Novel treatments using bee venom as immunotherapy 56 has also shown promise in clinical trials in cancer patients, 57 arthritis, 58 and other illnesses, and has been shown to disrupt Borrelia biofilm in vitro. ...
... The potential antimicrobial effects of Uncaria tomentosa have not been widely evaluated. In a non-peer reviewed publication, Uncaria tomentosa was reported to have antiborrelial effects in an in vitro model (115). Uncaria tomentosa has also been shown in peer reviewed research to have antimicrobial effects against human oral pathogens (116). ...
Full-text available
Lyme disease is the most common vector-borne disease in the US and Europe. Although the current recommended Lyme antibiotic treatment is effective for the majority of Lyme disease patients, about 10–20% of patients continue to suffer from persisting symptoms. There have been various anecdotal reports on the use of herbal extracts for treating patients with persisting symptoms with varying degree of improvements. However, it is unclear whether the effect of the herb products is due to their direct antimicrobial activity or their effect on host immune system. In the present study, we investigated the antimicrobial effects of 12 commonly used botanical medicines and three other natural antimicrobial agents for potential anti-Borrelia burgdorferi activity in vitro. Among them, 7 natural product extracts at 1% were found to have good activity against the stationary phase B. burgdorferi culture compared to the control antibiotics doxycycline and cefuroxime. These active botanicals include Cryptolepis sanguinolenta, Juglans nigra (Black walnut), Polygonum cuspidatum (Japanese knotweed), Artemisia annua (Sweet wormwood), Uncaria tomentosa (Cat's claw), Cistus incanus, and Scutellaria baicalensis (Chinese skullcap). In contrast, Stevia rebaudiana, Andrographis paniculata, Grapefruit seed extract, colloidal silver, monolaurin, and antimicrobial peptide LL37 had little or no activity against stationary phase B. burgdorferi. The minimum inhibitory concentration (MIC) values of Artemisia annua, Juglans nigra, and Uncaria tomentosa were quite high for growing B. burgdorferi, despite their strong activity against the non-growing stationary phase B. burgdorferi. On the other hand, the top two active herbs, Cryptolepis sanguinolenta and Polygonum cuspidatum, showed strong activity against both growing B. burgdorferi (MIC = 0.03–0.06% and 0.25–0.5%, respectively) and non-growing stationary phase B. burgdorferi. In subculture studies, only 1% Cryptolepis sanguinolenta extract caused complete eradication, while doxycycline and cefuroxime and other active herbs could not eradicate B. burgdorferi stationary phase cells as many spirochetes were visible after 21-day subculture. Further studies are needed to identify the active constituents of the effective botanicals and evaluate their combinations for more effective eradication of B. burgdorferi in vitro and in vivo. The implications of these findings for improving treatment of persistent Lyme disease are discussed.
... Publications in non-peer-reviewed publications on proprietary extracts of Uncaria suggest it might have some activity, but until more credible evidence is presented this should be viewed as extremely preliminary. 56 These herbs may have inflammation-modulating and other actions that might help in Lyme disease, and some do have evidence of efficacy against other infections organisms. So while the theoretical basis for their use may be as sound as any other extrapolations for other diseases, they remain theoretical. ...
Full-text available
Numerous tick-borne infections cause problems for humans and animals worldwide. Lyme disease is the best known, but babesiosis, bartonellosis, anaplasmosis, ehrlichiosis, Rocky Mountain spotted fever, and many others are also serious problems. Many studies have confirmed that herbs and herbal extracts can help to repel the several types of ticks (as can non-chemical means such as wearing long pants and tucking them into socks) that spread these diseases, as well as inhibit their reproduction. Corymbia citriodora (lemon eucalyptus) and its compound para-menthane-3,8-diol (PMD) have most convincingly been shown to be effective. 2-Undecanone and nootkatone with carvacrol have also shown some promise. Despite many claims of efficacy, no published clinical research could be located on any herb or herbal constituent to treat any tick-borne illness and thereby validate these claims. Dipsacus fullonum (fuller's teasel) is used as a case study to show a promising herb that has simply no research on whether it helps people with Lyme disease or any tick-borne infection. In vitro and animal studies show some promise for herbs and herbal compounds such as Brucea javanica (Java brucea) and artemisinin, but they have not been studied in clinical trials. There remains a black hole in terms of supported herbal treatments for these infections that urgently needs to be filled with credible clinical trials.
... In the presence of extract from Otoba parvifolia, the size of biofilm colonies was not reduced; however, bactericidal effect was achieved in >90%. In the presence of both herbal extracts, no sign of any colony formation was observed, with only a few nonmotile live spirochetes and rounded bodies present [Datar et al. 2010]. There is a need for more detailed toxicological evaluation of these extracts, although, the available data show no indication of severe toxicity and a low potential for acute and sub-acute oral toxicity, and there was no evidence of genotoxic or mutagenic activity [Valerio and Gonzales, 2005]. ...
Naturally occurring substances have been used for centuries to fight against various pathogens. They serve as a source for new chemical entities or provide options to already existing therapeutics. While there is an increasing interest in studying antimicrobial properties of naturally derived agents, little is known about their effects against Borrelia burgdorferi sensu lato, the causative pathogens of Lyme disease. A better understanding of this aspect could advance knowledge about pathophysiology of these bacteria and help improve the efficacy of current approaches against Lyme disease. Here, we review all naturally occurring substances scientifically evaluated to date, including plant extracts, their metabolites, and micronutrients, against vegetative (spirochetes) and latent (rounded bodies, biofilm) forms of Borrelia sp. This summary reveals the potent anti-borreliae activity of several of these natural compounds indicating their potential in enhancing the efficacy of current treatments for Lyme disease, and offering new options to already existing therapeutic regiments.
... Natural antimicrobial agents, which have been used for thousands of years, have been shown to be effective against various pathogens [32]. Many in vitro and clinical stud-ies have demonstrated their effectiveness not only against B. burgdorferi but also against many other pathogens [33][34][35][36][37][38]. Stevia rebaudiana which belongs to the Asteraceae family is typically referred to as honey leaf or sweet leaf, and due to its natural sweetness, it is used as a natural substitute to synthetic sweetener [39][40][41]. ...
Full-text available
Lyme disease is a tick-borne multisystemic disease caused by Borrelia burgdorferi. Administering antibiotics is the primary treatment for this disease; however, relapse often occurs when antibiotic treatment is discontinued. The reason for relapse remains unknown, but recent studies suggested the possibilities of the presence of antibiotic resistant Borrelia persister cells and biofilms. In this study, we evaluated the effectiveness of whole leaf Stevia extract against B. burgdorferi spirochetes, persisters, and biofilm forms in vitro. The susceptibility of the different forms was evaluated by various quantitative techniques in addition to different microscopy methods. The effectiveness of Stevia was compared to doxycycline, cefoperazone, daptomycin, and their combinations. Our results demonstrated that Stevia had significant effect in eliminating B. burgdorferi spirochetes and persisters. Subculture experiments with Stevia and antibiotics treated cells were established for 7 and 14 days yielding, no and 10% viable cells, respectively compared to the above-mentioned antibiotics and antibiotic combination. When Stevia and the three antibiotics were tested against attached biofilms, Stevia significantly reduced B. burgdorferi forms. Results from this study suggest that a natural product such as Stevia leaf extract could be considered as an effective agent against B. burgdorferi.
Au cours des dernières années, la borréliose de Lyme a de plus en plus fait parler d'elle. Connue depuis longtemps, il semblerait qu'elle cache encore bien des secrets et semble donner du fil à retordre aux autorités et aux professionnels de santé. Pathologie à la clinique complexe de par la variété de ses symptômes et sa ressemblance avec d'autres pathologies, la borréliose de Lyme, difficile à diagnostiquer autant qu'à traiter, semble bien répondre aux outils de la naturopathie. La prise en charge du patient dans sa globalité, le soutien de son organisme et ses défenses, l'apport de vitamines, minéraux et probiotiques, la modification des habitudes alimentaires et les outils de la phytothérapie peuvent constituer un bon soin d'accompagnement des outils de la médecine conventionnelle. Ce travail détaille la physiopathologie de la maladie puis les outils de la médecine conventionnelle face à elle avant d'aborder la place de la naturopathie dans la prise en charge de la borréliose de Lyme.
Full-text available
Uncaria tomentosa is considered a medicinal plant used over centuries by the peruvian population as an alternative treatment for several diseases. Many microorganisms usually inhabit the human oral cavity and under certain conditions can become etiologic agents of diseases. The aim of the present study was to evaluate the antimicrobial activity of different concentrations of Uncaria tomentosa on different strains of microorganisms isolated from the human oral cavity. Micropulverized Uncaria tomentosa was tested in vitro to determine the minimum inhibitory concentration (MIC) on selected microbial strains. The tested strains were oral clinical isolates of Streptococcus mutans, Staphylococcus spp., Candida albicans, Enterobacteriaceae and Pseudomonas aeruginosa. The tested concentrations of Uncaria tomentosa ranged from 0.25-5% in Müeller-Hinton agar. Three percent Uncaria tomentosa inhibited 8% of Enterobacteriaceae isolates, 52% of S. mutans and 96% of Staphylococcus spp. The tested concentrations did not present inhibitory effect on P. aeruginosa and C. albicans. It could be concluded that micropulverized Uncaria tomentosa presented antimicrobial activity on Enterobacteriaceae, S. mutans and Staphylococcus spp. isolates.
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The effectiveness of a new first-in-class antibiotic, tigecycline (glycylcycline), was evaluated during the early dissemination (1 week), early immune (3 weeks), or late persistent (4 months) phases of Borrelia burgdorferi infection in C3H mice. Mice were treated with high or low doses of tigecycline, saline (negative-effect controls), or a previously published regimen of ceftriaxone (positive-effect controls). Infection status was assessed at 3 months after treatment by culture, quantitative ospA real-time PCR, and subcutaneous transplantation of joint and heart tissue into SCID mice. Tissues from all saline-treated mice were culture and ospA PCR positive, tissues from all antibiotic-treated mice were culture negative, and some of the tissues from most of the mice treated with antibiotics were ospA PCR positive, although the DNA marker load was markedly decreased compared to that in saline-treated mice. Antibiotic treatment during the early stage of infection appeared to be more effective than treatment that began during later stages of infection. The viability of noncultivable spirochetes in antibiotic-treated mice (demonstrable by PCR) was confirmed by transplantation of tissue allografts from treated mice into SCID mice, with dissemination of spirochetal DNA to multiple recipient tissues, and by xenodiagnosis, including acquisition by ticks, transmission by ticks to SCID mice, and survival through molting into nymphs and then into adults. Furthermore, PCR-positive heart base tissue from antibiotic-treated mice revealed RNA transcription of several B. burgdorferi genes. These results extended previous studies with ceftriaxone, indicating that antibiotic treatment is unable to clear persisting spirochetes, which remain viable and infectious, but are nondividing or slowly dividing.
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Persistence of tissue spirochetes of Borrelia burgdorferi as helices and round bodies (RBs) explains many erythema-Lyme disease symptoms. Spirochete RBs (reproductive propagules also called coccoid bodies, globular bodies, spherical bodies, granules, cysts, L-forms, sphaeroplasts, or vesicles) are induced by environmental conditions unfavorable for growth. Viable, they grow, move and reversibly convert into motile helices. Reversible pleiomorphy was recorded in at least six spirochete genera (>12 species). Penicillin solution is one unfavorable condition that induces RBs. This antibiotic that inhibits bacterial cell wall synthesis cures neither the second "Great Imitator" (Lyme borreliosis) nor the first: syphilis. Molecular-microscopic techniques, in principle, can detect in animals (insects, ticks, and mammals, including patients) helices and RBs of live spirochetes. Genome sequences of B. burgdorferi and Treponema pallidum spirochetes show absence of >75% of genes in comparison with their free-living relatives. Irreversible integration of spirochetes at behavioral, metabolic, gene product and genetic levels into animal tissue has been documented. Irreversible integration of spirochetes may severely impair immunological response such that they persist undetected in tissue. We report in vitro inhibition and destruction of B. burgdorferi (helices, RBs = "cysts") by the antibiotic Tigecycline (TG; Wyeth), a glycylcycline protein-synthesis inhibitor (of both 30S and 70S ribosome subunits). Studies of the pleiomorphic life history stages in response to TG of both B. burgdorferi and Treponema pallidum in vivo and in vitro are strongly encouraged.
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The long latent stage seen in syphilis, followed by chronic central nervous system infection and inflammation, can be explained by the persistence of atypical cystic and granular forms of Treponema pallidum. We investigated whether a similar situation may occur in Lyme neuroborreliosis. Atypical forms of Borrelia burgdorferi spirochetes were induced exposing cultures of Borrelia burgdorferi (strains B31 and ADB1) to such unfavorable conditions as osmotic and heat shock, and exposure to the binding agents Thioflavin S and Congo red. We also analyzed whether these forms may be induced in vitro, following infection of primary chicken and rat neurons, as well as rat and human astrocytes. We further analyzed whether atypical forms similar to those induced in vitro may also occur in vivo, in brains of three patients with Lyme neuroborreliosis. We used immunohistochemical methods to detect evidence of neuroinflammation in the form of reactive microglia and astrocytes. Under these conditions we observed atypical cystic, rolled and granular forms of these spirochetes. We characterized these abnormal forms by histochemical, immunohistochemical, dark field and atomic force microscopy (AFM) methods. The atypical and cystic forms found in the brains of three patients with neuropathologically confirmed Lyme neuroborreliosis were identical to those induced in vitro. We also observed nuclear fragmentation of the infected astrocytes using the TUNEL method. Abundant HLA-DR positive microglia and GFAP positive reactive astrocytes were present in the cerebral cortex. The results indicate that atypical extra- and intracellular pleomorphic and cystic forms of Borrelia burgdorferi and local neuroinflammation occur in the brain in chronic Lyme neuroborreliosis. The persistence of these more resistant spirochete forms, and their intracellular location in neurons and glial cells, may explain the long latent stage and persistence of Borrelia infection. The results also suggest that Borrelia burgdorferi may induce cellular dysfunction and apoptosis. The detection and recognition of atypical, cystic and granular forms in infected tissues is essential for the diagnosis and the treatment as they can occur in the absence of the typical spiral Borrelia form.
Minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) were evaluated of the antimicrobial chemotherapeutics amoxicillin, azithromycin, cefotaxime, ceftriaxone, doxycycline, penicillin G sodium, roxithromycin, and trimethoprim-sulfamethoxazole for 30 Borrelia strains from various sources (skin, cerebrospinal fluid, ticks). Of these strains 29 were Lyme disease agents of the species Borrelia afzelii (n = 12), Borrelia burgdorferi sensu stricto (n = 4), Borrelia garinii (n = 13), and one was the relapsing fever strain Borrelia turicatae (n = 1). Tests were performed in microtiter plates by broth dilution. MIC was determined after 72 hours of incubation by comparing growth control with the antibiotic dilutions by means of dark field microscopy. Strains tested were susceptible against amoxicillin, azithromycin, cefotaxime, ceftriaxone, doxycycline, and penicillin G sodium, partly susceptible to roxithromycin, and resistant to trimethoprim-sulfamethoxazole. No statistically significant differences in MIC and MBC were seen among the different antibiotics with the various Borrelia species.
The purpose of this study was to examine the structural alterations of Borrelia burgdorferi when exposed to spinal fluid. Normal, mobile spirochetes were inoculated into spinal fluid, and the spirochetes were converted to cysts (spheroplast L-forms) after 1-24 h. When these cystic forms were transferred to a rich BSK-H medium, the cysts were converted back to normal, mobile spirochetes after incubation for 9 to 17 days. The cultures were examined by dark field microscopy (DFM), interference contrast microscopy (ICM) and transmission electron microscopy (TEM). When neuroborreliosis is suspected, it is necessary to realize that B. burgdorferi can be present in a cystic form, and these cysts have to be recognized by microscopy. This study may also explain why cultivation of spinal fluid often is negative with respect to B. burgdorferi.
Recently, Uncaria tomentosa (Willd.) D.C. has become known as a healing plant with an ethnomedicinal background. There have been several reports on its constituents, in particular, oxindole alkaloids. It was found that 2 chemotypes of Uncaria tomentosa with different alkaloid patterns occur in nature. The roots of one type contain pentacyclic oxindoles and the other contains tetracyclic oxindoles. This difference should be considered when the plant is to be used for medicinal applications. Tetracyclic oxindole alkaloids act on the central nervous system, whereas pentacyclic oxindole alkaloids affect the cellular immune system. Recent studies have shown that the tetracyclic alkaloids exert antagonistic effects on the action of the pentacyclic alkaloids. Mixtures of these 2 types of drugs are therefore unsuitable for medicinal uses.
Background It is controversial whether prolonged antibiotic treatment is effective for patients in whom symptoms persist after the recommended antibiotic treatment for acute Lyme disease. Methods We conducted two randomized trials: one in 78 patients who were seropositive for IgG antibodies to Borrelia burgdorferi at the time of enrollment and the other in 51 patients who were seronegative. The patients received either intravenous ceftriaxone, 2 g daily for 30 days, followed by oral doxycycline, 200 mg daily for 60 days, or matching intravenous and oral placebos. Each patient had well-documented, previously treated Lyme disease but had persistent musculoskeletal pain, neurocognitive symptoms, or dysesthesia, often associated with fatigue. The primary outcome measures were improvement on the physical- and mental-health–component summary scales of the Medical Outcomes Study 36-Item Short-Form General Health Survey (SF-36) — a scale measuring the health-related quality of life — on day 180 of the study. Results After a planned interim analysis, the data and safety monitoring board recommended that the studies be discontinued because data from the first 107 patients indicated that it was highly unlikely that a significant difference in treatment efficacy between the groups would be observed with the planned full enrollment of 260 patients. Base-line assessments documented severe impairment in the patients' health-related quality of life. In intention-to-treat analyses, there were no significant differences in the outcomes with prolonged antibiotic treatment as compared with placebo among either the seropositive or the seronegative patients. Conclusions There is considerable impairment of health-related quality of life among patients with persistent symptoms despite previous antibiotic treatment for acute Lyme disease. However, in these two trials, treatment with intravenous and oral antibiotics for 90 days did not improve symptoms more than placebo.
Cystic forms (also called spheroplasts or starvation forms) and their ability to reconvert into normal motile spirochetes have already been demonstrated in the Borrelia burgdorferi sensu lato complex. The aim of this study was to determine whether motile B. garinii could develop from cystic forms, not only in vitro but also in vivo, in cyst-inoculated mice. The cysts prepared in distilled water were able to reconvert into normal motile spirochetes at any time during in vitro experiments, lasting one month, even after freeze-thawing of the cysts. Motile spirochetes were successfully isolated from 2 out of 15 mice inoculated intraperitoneally with cystic forms, showing the infectivity of the cysts. The demonstrated capacity of the cysts to reconvert into motile spirochetes in vivo and their surprising resistance to adverse environmental conditions should lead to further studies on the role and function of these forms in Lyme disease.
In our search for therapeutical alternatives for antiprotozoal chemotherapy, we collected a selection of 44 plants from western Colombia upon ethnopharmacological and chemotaxonomic considerations. Polar and apolar extracts of these species were examined for antimalarial activity using in vitro tests with two clones of Plasmodium falciparum. Leishmanicidal and trypanocidal activity were determined in vitro using promastigote and amastigote forms of several strains of Leishmania sp. and epimastigotes of Trypanosoma cruzi. Among the selected plants, the 15 following species showed good or very good antiprotozoal activity in vitro: Aspidosperma megalocarpon, Campnosperma panamense, Conobea scoparioides, Guarea polymera, Guarea guidonia, Guatteria amplifolia, Huberodendron patinoi, Hygrophila guianensis, Jacaranda caucana, Marila laxiflora, Otoba novogranatensis, Otoba parviflora, Protium amplium, Swinglea glutinosa and Tabernaemontana obliqua. Cytotoxicity was assessed in U-937 cells and the ratio of cytotoxicity to antiprotozoal activity was determined for the active extracts. Ten extracts from eight species showed selectivity indexes > or = 10. Among the extracts that showed leishmanicidal activity, the methylene chloride extract of leaves from C. scoparioides showed a selectivity index in the same range that the one of the Glucantime control. Several of the active leishmanicidal plants are traditionally used against leishmaniasis by the population of the concerned area.