Culture of the Entire Mouse to Determine Persistance of c.BB

Topics with information and discussion about published studies related to Lyme disease and other tick-borne diseases.
duncan
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Re: Culture of the Entire Mouse to Determine Persistance of

Post by duncan » Thu 4 Sep 2014 17:40

hv808ct, the inference then is that the VBNC form would first have to be identified, yes? I guess that would be the crux of the problem.

What is it that Kim Lewis was finding? What form were those persister cells in the .001% or whatever that survived post-treatment?

Not sure if that question is relevant.

dlf
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Re: Culture of the Entire Mouse to Determine Persistance of

Post by dlf » Fri 5 Sep 2014 2:41

It would appear that this experiment wasn't designed to determine whether there were VBNC forms which could by some other method become cultivable, it was simply to try to establish whether the inability to culture the bacteria after antibiotics using conventional methods might be the result of too few cultivable cells.
The liquid contents (about 1.0 ml) of the minced body parts were collected and then injected intraperitoneally into separate normal recipient SKH mice. Four weeks later, the recipient mice were euthanized and separate extracts of excised urinary bladders and ears were prepared, cultured, and examined microscopically as described above; positive cultures were then subcultured….
I found it interesting that Dr. Wormser injected the liquified antibiotic treated mice into fresh naive mice. Maybe he thought that these new mice could act as a culture medium in place of BSK at least as a starter, but if that is the case why only wait one month when the Hodzic et al study didn't show resurgence until 12 months later?

Besides, if he actually wanted to culture the spirochetes after antibiotic treatment and thereby show that it could be done, he doesn't need to re-invent the wheel. This has already been accomplished. He just needed to treat the mice with anti-tumor necrosis factor alpha. An interesting experiment would have been to treat the naive mice with the anti-TNF-a and see if that would have worked as well.

http://jid.oxfordjournals.org/content/1 ... 9.full.pdf

• JID 2007:195 (15 May) • Yrja¨na¨inen et al.
Anti–Tumor Necrosis Factor–a Treatment
Activates Borrelia burgdorferi Spirochetes 4
Weeks after Ceftriaxone Treatment in C3H/He Mice


Background. The effect of anti–tumor necrosis factor (TNF)–a treatment in Borrelia burgdorferi–infected and
ceftriaxone-treated C3H/He mice was evaluated.
Methods. Mice were infected with B. garinii A˚ 218 or B. burgdorferi sensu stricto N40. At 2 weeks of infection,
one group was treated simultaneously with ceftriaxone and anti–TNF-a, whereas another received ceftriaxone at
2 weeks and anti–TNF-a 4 weeks later. One group received ceftriaxone treatment only. Infected and noninfected
control groups were sham treated.
Results. At 14 weeks of infection, B. burgdorferi could not be detected by cultivation or by polymerase chain
reaction in tissue samples of any mouse treated with ceftriaxone only. However, spirochetes grew from the tissue
samples of one-third of the mice treated with anti–TNF-a simultaneously or 4 weeks after ceftriaxone. These
activated spirochetes showed ceftriaxone sensitivity rates, plasmid profiles, and virulence rates similar to those of
bacteria used to infect the mice. All infected control mice and mice given anti–TNF-a only were culture positive.
Conclusions. This report shows that, after ceftriaxone treatment for 5 days, a portion of B. burgdorferi–infected
mice still have live spirochetes in their body, which are activated by anti–TNF-a treatment.
The link above is to the full study article and it is an interesting read. This makes me wonder whether the level of TNF-a gradually drops after the antibiotic treatment and after about 12 months (at least in the murine model) it is sufficiently low that the spirochetes are able to resurge.

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ChronicLyme19
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Re: Culture of the Entire Mouse to Determine Persistance of

Post by ChronicLyme19 » Fri 5 Sep 2014 3:51

Ok, stuipid Q, for what you are suggesting for mice, wouldn't' the imply that you'd want to give people anti-TNF-a, to get the bugs to emerge from their hiding spots and then slam them with abx?
Half of what you are taught is incorrect, but which half? What if there's another half missing?

hv808ct
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Re: Culture of the Entire Mouse to Determine Persistance of

Post by hv808ct » Fri 5 Sep 2014 4:20

Re: Culture of the Entire Mouse to Determine Persistance of
Postby dlf » Fri 5 Sep 2014 2:41
The link above is to the full study article and it is an interesting read. This makes me wonder whether the level of TNF-a gradually drops after the antibiotic treatment and after about 12 months (at least in the murine model) it is sufficiently low that the spirochetes are able to resurge.
Read the paper's comments at: http://jid.oxfordjournals.org/content/1 ... 866.2.long

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ChronicLyme19
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Re: Culture of the Entire Mouse to Determine Persistance of

Post by ChronicLyme19 » Fri 5 Sep 2014 4:50

Hot damn.
Unexpectedly, some of the mice treated with ceftriaxone and anti-TNF-α turned out to be culture-positive for B. burgdorferi, whereas all animals treated with ceftriaxone only were culture negative (table 1 in our article [3]). Most importantly, 3 of 10 mice that received anti-TNF-α 4 weeks after ceftriaxone were culture positive for B. burgdorferi. This finding suggests that it is not a question of anti-TNF-α being “able to rescue B. burgdorferi from the bactericidal effect of ceftriaxone,” as stated by Wormser et al. [2, pp. 1865] in their correspondence, but rather that B. burgdorferi spirochetes are able to escape the killing effect of ceftriaxone. We find these results extremely interesting and worth reporting.
Ceftriaxone-treated mice have only had positive culture results if they have received both ceftriaxone and anti-TNF-α. Thus, we believe that this phenomenon may have biological significance.
So the lyme was not totally gone in some mice, just in hiding/not in a cultivable state until triggered.
Half of what you are taught is incorrect, but which half? What if there's another half missing?

lou
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Re: Culture of the Entire Mouse to Determine Persistance of

Post by lou » Sat 6 Sep 2014 2:28

So if you don't want to find spirochetes in your culture, make sure not to give anti-TNF.

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