Sapi - Antibiotic susceptibility different forms of Bb

Topics with information and discussion about published studies related to Lyme disease and other tick-borne diseases.
duncan
Posts: 1370
Joined: Wed 5 Sep 2012 18:48

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by duncan » Fri 22 May 2015 17:31

Two observations:

1) 90% effective might as well be not effective at all; spirochetes tend to reproduce, so all you need is less than 1%. An assumption may be that the human immune system will account for the remainder; that's an assumption arguably made at patients' expense.

2) All this is in vitro; extrapolating to inside the human body is ill-advised. But at least Sapi et al make a genuine scientific effort to demonstrate why patients remain ill with Bb.

User avatar
ChronicLyme19
Posts: 564
Joined: Mon 11 Aug 2014 17:42
Location: NY, USA

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by ChronicLyme19 » Fri 22 May 2015 17:43

Martian wrote: But when the number of cystic round body forms increased about twofold (shouldn't that be threefold?), what does that mean in terms of numbers? Is it an increase from 10 to 30, or from 100 to 300?

If the numbers of cystic round body forms were very low to begin with, which I think is probable, then maybe the increase to 300% percent of the control isn't so dramatic as it would seem at first sight.
Good question, I see all the charts in percentages. Although any live form increasing with added antibiotics concerns me. Doxycycline was the only one that showed this effect compared to the other antibiotics studied. The study also showed tinidazole to be just as effective as doxycycline, but without any increase in round body forms, so that seems like a better option. Not to mention the lack of biofilm formation.
Tinidazole.jpg
Tinidazole.jpg (136.11 KiB) Viewed 1793 times
Caption 1
Susceptibility of the spirochete and round body forms of strain B31 (top panels) and strain S297 (bottom panels) of B. burgdorferi to the most effective concentrations of three antibiotics measured by dark-field microscopy. Tinidazole, metronidazole, and doxycycline effect on B. burgdorferi after 3 weeks of subculturing following 72-hour treatment.

Note: *P values <0.05 indicates statistical significance compared with control.
Biofilms.jpg
Biofilms.jpg (186.35 KiB) Viewed 1793 times
Caption 2
Evaluation of biofilm-like colonies of B. burgdorferi. Qualitative analysis of biofilm-like colonies of strain B31 measured by fluorescent microscopy using SYTO®9 green-fluorescent stain (live organisms) and propidium iodide red-fluorescent stain (dead organisms): (Ba) Control; (Bb) Doxycycline; (Bc) Tinidazole; (Bd) Tigecycline; (Be) Metronidazole; (Bf) Amoxicillin.
Half of what you are taught is incorrect, but which half? What if there's another half missing?

Martian
Posts: 1944
Joined: Thu 26 Jul 2007 18:29
Location: Friesland, the Netherlands

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by Martian » Fri 22 May 2015 19:24

ChronicLyme19 wrote:Good question, I see all the charts in percentages. Although any live form increasing with added antibiotics concerns me. Doxycycline was the only one that showed this effect compared to the other antibiotics studied. The study also showed tinidazole to be just as effective as doxycycline, but without any increase in round body forms, so that seems like a better option. Not to mention the lack of biofilm formation.
Tinidazole is an interesting drug, as was shown in previous studies. I wished they had also included hydroxychloroquine (Plaquenil), which is another known "cyst buster", at least in vivo. But the question is how 72 hours at concentrations above the calculated MIC and MBC in a test tube translates to patients who take the drugs for weeks/months. Just one thing to take into account are the concentrations reached throughout the body with acceptable dosages, and there are several more factors. One needs to be very careful with drawing conclusions from such in vitro studies.

A specific note about Metronidazole (may count for Tinidazole as well):

Source: http://www.antibiotics-info.org/metronidazole.html
Mechanism of action

Metronidazole is not active by itself; its nitro group gets activated to convert it into the active form. This activation is done by the micro-organisms. It is selectively toxic to anaerobic and micro-aerophilic pathogens. After entering the cell by diffusion its nitro group is reduced by certain redox proteins functional only in anaerobic microorganisms to extremely reactive nitrogen radical that is cytotoxic. The nitro radical of metronidazole acts as an electron sink which competes with the biological electron acceptors of the anaerobic organism for the electrons produced by the pyruvate: ferrodoxin Oxidoreductase (PFOR) enzyme pathway of pyruvate oxidation. The energy metabolism of anaerobes is, thus, disturbed and replication, transcription and repair process of DNA is hindered leading to cell death. Metronidazole and oxygen both strive for the electrons formed during metabolism of energy. Hence oxygen decreases the cytotoxic action caused by metronidazole and also reduces its activation. Anaerobes which develop metronidazole resistance become deficient in the mechanism that generates the reactive nitro radical from it.
A German research article stated:

Source: http://www.lymeneteurope.org/forum/view ... p=127#p129
under in vitro conditions Metronidazol is not converted into its active form (because NADPH Nitroreductase enzyme is not available).

edit: fixed text error.
Last edited by Martian on Fri 22 May 2015 21:07, edited 1 time in total.

RitaA
Posts: 2768
Joined: Thu 1 Jul 2010 8:33

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by RitaA » Fri 22 May 2015 20:06

A European research team has studied the in vitro susceptibility of 9 human isolates of European Borrelia burgdorferi sensu stricto to 6 antibiotics. Unfortunately, only the article abstract is currently available to the general public:

http://www.ncbi.nlm.nih.gov/pubmed/23312603
Int J Antimicrob Agents. 2013 Mar;41(3):288-91. doi: 10.1016/j.ijantimicag.2012.11.016. Epub 2013 Jan 9.

In vitro susceptibility of European human Borrelia burgdorferi sensu stricto strains to antimicrobial agents.

Veinović G, Cerar T, Strle F, Lotrič-Furlan S, Maraspin V, Cimperman J, Ružić-Sabljić E.

Source

Department of Microbiology and Immunology, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11221 Belgrade, Serbia.

Abstract

Broth microdilution and macrodilution assays were used to determine minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of six antimicrobial agents (ceftriaxone, cefuroxime sodium, azithromycin, amoxicillin, doxycycline and amikacin) for nine European human isolates of Borrelia burgdorferi sensu stricto (s.s.). Strains were obtained from patients diagnosed with Lyme borreliosis in Slovenia. Modified Kelly-Pettenkofer medium with a final inoculum of 10(5)Borrelia cells/mL and incubation periods of 72h and of 3 weeks and 6 weeks were used in the determination of MICs and MBCs, respectively. Observed MICs indicated that all isolates were susceptible to all the tested antimicrobial agents with the exception of amikacin. Cefuroxime sodium (MIC(90)=0.063mg/L), azithromycin (MIC(90)=0.22mg/L) and ceftriaxone (MIC(90)=0.25mg/L) displayed the lowest MICs, followed by amoxicillin (MIC(90)=1mg/L) and doxycycline (MIC(90)=2mg/L); no strain was susceptible to amikacin (MIC(90)=256mg/L). MBCs after incubation for 3 weeks and 6 weeks were determined for amoxicillin (MBC(90)=32mg/L), doxycycline (MBC(90)=32mg/L) and amikacin (MBC(90)=1024mg/L) and were found to be high (but not defined) for azithromycin (MBC(90)>0.88mg/L), cefuroxime sodium (MBC(90)>4mg/L) and ceftriaxone (MBC(90)>4mg/L). In determination of borrelial susceptibility to antimicrobial agents, intrinsic low susceptibility or methodological factors could result in low in vitro susceptibility of individual strains. This study is the first report on the antibiotic susceptibility of a series of European human isolates of B. burgdorferi s.s.

Copyright © 2013 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

PMID: 23312603 [PubMed - in process]
The full article can be purchased for $35.95:

http://www.sciencedirect.com/science/ar ... 7912004530

nnecker
Posts: 215
Joined: Wed 19 Dec 2012 22:57

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by nnecker » Fri 22 May 2015 22:16

Here are some in vivo round bodies:

http://www.jci.org/articles/view/58813/figure/2

I'm sure some of these "cysts" were mix in the mouse material that was prepared for culture in BSK.Of course the cultures were always negative.

duncan
Posts: 1370
Joined: Wed 5 Sep 2012 18:48

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by duncan » Fri 22 May 2015 22:30

Outside of an EM, culturing is a joke. nnecker, if you are going to comment, it would be nice if you commented fully and fairly and not selectively: Although culture attempts were unsuccessful in this particular effort you've provided a link to, spirochetes - post-treatment! - were recovered through xenodiagnosis. And this is a Bockenstedt effort...

User avatar
ChronicLyme19
Posts: 564
Joined: Mon 11 Aug 2014 17:42
Location: NY, USA

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by ChronicLyme19 » Tue 26 May 2015 15:45

Martian wrote: Tinidazole is an interesting drug, as was shown in previous studies. I wished they had also included hydroxychloroquine (Plaquenil), which is another known "cyst buster", at least in vivo. But the question is how 72 hours at concentrations above the calculated MIC and MBC in a test tube translates to patients who take the drugs for weeks/months. Just one thing to take into account are the concentrations reached throughout the body with acceptable dosages, and there are several more factors. One needs to be very careful with drawing conclusions from such in vitro studies.
I agree, but you have to start somewhere. The only studies I have found so far on Plaquenil or grapefruit seed extract on round body busting is from Brorson, but I don't have acess to the full studies. to see if they are any good.

http://www.ncbi.nlm.nih.gov/pubmed/12102233
http://www.ncbi.nlm.nih.gov/pubmed/17565468
Half of what you are taught is incorrect, but which half? What if there's another half missing?

X-member
Posts: 3954
Joined: Mon 30 Jul 2007 18:18

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by X-member » Sun 8 Nov 2015 0:55

Martian wrote earlier in this thread:
If the numbers of cystic round body forms were very low to begin with, which I think is probable, then maybe the increase to 300% percent of the control isn't so dramatic as it would seem at first sight.
ILADS 2015: From Bench to Bedside: Translating Research Into Clinical Practice

http://www.betterhealthguy.com/ilads-2015

A quote:
Leona Gilbert PhD spoke on "Morphological and Biochemical Features of Borrelia Burgdorferi Pleomorphic Forms" and shared:
- Pleomorphism allows for the evasion of the immune system and is a consequence of persistent conditions.
- There are coccoid, rod, spirochete, and biofilm forms - two or more forms is pleomorphism.
- In Lyme, there are spirochetes, blebs and loops, round bodies, and biofilms.
- A spirochete averages 20 microns; a round body 3 microns.
- A decrease in spirochetes leads to an increase in blebs and round bodies.
- Human serum induces round bodies.
- Round bodies can go back to spirochetal forms.
- Round bodies have no metabolic activity but they do when they convert back to spirochetes.
- Round bodies are not "cysts".

X-member
Posts: 3954
Joined: Mon 30 Jul 2007 18:18

Re: Sapi - Antibiotic susceptibility different forms of Bb

Post by X-member » Wed 6 Jul 2016 23:47

Bump!

Post Reply