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» LymeNet Flash » Questions and Discussion » Medical Questions » Low Cost way to see Lyme in blood.

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Author Topic: Low Cost way to see Lyme in blood.
D Bergy
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It is not out of the question to see Lyme without being a lab tech, here is an average person in the U.K. that shows how it is done.

http://www.lyme-diagnosis.org.uk/

D Bergy

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tcw
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I think that one of three things is happening:

What is being seen on the scope is not Bb.

This person has a monumental amount of Bb in their plasma.

The science of PCR testing is completely useless and we are all mislead. A random drop of blood that has visible Bb would light up a PCR with near certainty.

I have a hard time believing that what is pictured is Bb. If Bb was that easy to see in plasma then the testing dilemma would be almost non-existent, a positive PCR on that blood would be pretty much guaranteed.

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Eight Legs Bad
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quote:
Originally posted by tcw:

A random drop of blood that has visible Bb would light up a PCR with near certainty.

No, it wouldnt. Or rather, it only would,if the testing lab used the correct primers.

Primers are short sequences of DNA which are specific to the organism and the strain that you are looking for. If the primers match the DNA in your test sample, they will facilitate the production of multiple copies of that genetic material, and it then becomes very easy to identify the bug - even if there was only a tiny number of bugs in the original sample.

However, if you use the wrong primers - perhaps because the patient has a strain of borrelia that differs somewhat from the one from which you obtained the primers - you may not get the match.

I know the engineer who set up that website www.lyme-diagnosis.org.uk ; some of the videos on the site relate to his own blood. He was PCR positive for Borrelia burgdorferi at the time.

Please also google for information about the work of Dr. Marie Kroun, who has many videos of darkfield examination of Lyme patients' blood on her site, done by various doctors. Some of the testing featured there shows confirmation with fluorescent antibody. This is extremely specific and proves beyond a doubt that the microbes seen are not artefacts, but borrelia.

The public health apparatus in the US, Britain and other countries do not want to use darkfield examination of blood, even though it was the standard for over 100 years for diagnosis of relapsing fever, another borrelia infection. It is still used to diagnose that borreliosis today.

They don't want it used because they are denying the persistence of Lyme, the existence and significance of the L-forms etc.. Primers to Bb sensu stricto strain B31 may well be irrelevant to many chronically ill patients.

The borrelia causing the "Lyme-like illness" in the South of the US, for example, is genetically different from Bb and will not register on a Borrelia burgdorferi PCR.

There is a blanket denial of the true prevalence of Lyme and related infection, its chronicity and the presence of the borrelia micro-organism in the blood. Part of the reason for this has been highlighted in the film "Under our Skin" (ie the interests of the powerful insurance and other industries), but a lot of it is due to other reasons which I've written about on my website
www.lyme-rage.info

Elena Cook

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MariaA
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one of the issues though is that for many if not most patients, Lyme doesn't really turn up in the blood. I know there are some cases where that's the case, but it's one reason that animal studies whereby the animal is autopsied at the end of the study can turn up lots of borrellia that are visible under a microscope, whereas fluid samples such as blood and CSF dont' do so in a human patient.

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treepatrol
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Once a animal dies and then the autopsie for Bb occurs its no longer in its spirochete form its either in cyst or bleb or coccoid forms because the temp it likes is 37 degrees celsius or 98.6 Fahrenheit amazing huh exactly what humans run at.

Deffinatly makes me wonder how a germ that was around so long just happens to like our temp the best?Hahahahaha

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Eight Legs Bad
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quote:
Originally posted by MariaA:
one of the issues though is that for many if not most patients, Lyme doesn't really turn up in the blood.

Most of the studies claiming that lyme borrelia are not found in the blood were done by the same Steere camp researchers who publish false information to the effect that Lyme is easily cured in a few weeks' antibiotics, almost never goes chronic etc..

The life cycle of tick-borne borrelia infection requires that it somehow gets back into the tick. If the borreliae did not appear in the blood of their host, human or animal, how would the parasites ever continue their natural life cycle?

They would die out in the body of the host and Lyme borreliosis would be extinct rather than a rapidly-growing epidemic.

Elena

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tcw
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quote:
No, it wouldnt. Or rather, it only would,if the testing lab used the correct primers.
True enough, but I believe that commercial kits with primers for reactivity for Bb sensu latu are available, but who knows what lab uses which primers. I think the primers are based on a highly conserved region of the flagellin gene.

quote:
The borrelia causing the "Lyme-like illness" in the South of the US, for example, is genetically different from Bb and will not register on a Borrelia burgdorferi PCR
Agreed, but the website specifically references Bb, not B. hermsii, lonestari, etc.

quote:
The public health apparatus in the US, Britain and other countries do not want to use darkfield examination of blood, even though it was the standard for over 100 years for diagnosis of relapsing fever, another borrelia infection. It is still used to diagnose that borreliosis today.

They don't want it used because they are denying the persistence of Lyme, the existence and significance of the L-forms etc..

I agree on the resistance to microscopy, but I am skeptical about the reasons why. Microscopy requires a highly trained and meticulous technician to produce accurate results, and even then I would guess that sensitivity would vary tremendously - everybody has bad days at work.

There is definitely a lot of denial going on, but at this point I am still in the camp of "Never attribute to malice that which can be attributed to stupidity".

On a more personal note, I do hope that this individual is being treated and making some progress on eradicating the infection.

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Eight Legs Bad
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quote:
Originally posted by tcw:
... I think the primers are based on a highly conserved region of the flagellin gene. ...

The primers used by which lab/labs?

The Steerites often refer in their papers to PCR done with primers based on OspA. It has been known for years that OspA varies widely between species (it's one of the reasons the LymeRix vaccine was never marketed in Europe) and so would seem a poor choice.

You seem to have a lot of knowledge about these matters - are you a microbiologist by any chance?

Elena

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nenet
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--------------------------------------------------
quote:"Eight Legs Bad
--------------------------------------------------
quote: Originally posted by MariaA:
one of the issues though is that for many if not most patients, Lyme doesn't really turn up in the blood.
--------------------------------------------------

Most of the studies claiming that lyme borrelia are not found in the blood were done by the same Steere camp researchers who publish false information to the effect that Lyme is easily cured in a few weeks' antibiotics, almost never goes chronic etc..

The life cycle of tick-borne borrelia infection requires that it somehow gets back into the tick. If the borreliae did not appear in the blood of their host, human or animal, how would the parasites ever continue their natural life cycle?

They would die out in the body of the host and Lyme borreliosis would be extinct rather than a rapidly-growing epidemic.

Elena"
------------------------------------------------

I have a specific interest in the issue of Lyme probability in blood, because I read something a while back that intrigued me. It was regarding the tick saliva compounds and their effect on the spirochetes in the host.

It stated that when a tick bites a host and releases its saliva, the complex chemical compound enters the host's bloodstream, and certain chemicals then act as a calling card to the spirochete, so that it leaves its deeper tissue quarters and enters the bloodstream to hitch a ride in the tick.

I wish I had a citation for you, but I can try to dig that up if needed. I imagine you have run across this before though. Are you aware of any studies corroborating this? Does this fall in line with your knowledge base? I am not married to any set ideas, just looking for the scientific truth as it can be discerned.

I am slow-going in my Lyme research, as I have neuroborreliosis symptoms that get in the way too often, but I am certainly interested. I don't want to divert the thread but hopefully this can be discussed in tandem...

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MariaA
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That info is quoted in Buhner's book- I don't have a copy in front of me but there's probably a reference to a study in the back, most of the book is written that way.

I don't know, I don't believe the 'it's all the Steerites' part, Eight Legs Bad. If it were so easy to find borrellia in the bloodstream then there wouldn't be anywhere near the difference between how human studies and animal studies are conducted and what they show.

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Eight Legs Bad
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quote:
Originally posted by MariaA:
If it were so easy to find borrellia in the bloodstream then there wouldn't be anywhere near the difference between how human studies and animal studies are conducted and what they show.

Well, you could say the same about any wrong idea which has been accepted by the medical mainstream. You could say, "if Lyme was really a chronic neurological disease, you wouldnt have all these studies showing it was an easily-cured infection that mostly affects the joints."

Once a wrong idea becomes an entrenched, accepted part of contemporary "knowledge" about an illness, researchers are less likely to conduct a study to disprove the accepted idea. Also, where would the funding come from?

Have a look at this extract below. The study's findings were not acknowledged at the time; however, today we know that ACA (a late maifestation) is caused by Lyme borreliosis.


Klin Wochenschr 1955;33:185−186
[Tierexperimentielle Untersuchungen zur tiologie der acrodermatitis chronica
atrophican Herxheimer]
Lohel H

"Citations translated from German:
Blood from patients with different dermatosis injected into mice, and the mice
were sacrificed after 14 days and tested for pallida−reaction (Pallida−antigen,
Promonta−Hamburg).
As shown in table 1, 58,95% of the ACA reacted positive in pallida−reaction,
while mice inoculated with blood from patients with other dermatosis were below
2% positive.
These results indicate an infectious etiology to ACA and point to a spirochete.
Most remarkable is that the infection could be tranferred by blood."

I don't have Buhner's book so I'm not sure which study you were referring to.

Elena

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Eight Legs Bad
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Sorry, I meant to clarify - the mice in the 1955 study I cited, which had been injected with the blood of ACA patients, were positive in "pallida-reaction". This means they were cross-reacting with a test for syphilis antibody.

Elena

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david1097
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I think it is follish not to think the bacteria is in the blood. Ticks do not eat tissue, they eat blood so I think it is pretty sure bet to say that blood is the carrier, at least in the pre-treatment stages. Where it is, I do not know, but for sure its in the blood.

Also remeber that lyme was discovered by seeing the spirocete in the tick hemolymph. It must therefore be in spiroceete form while in the insect.

All this being said, I do not agree with the conclusions drawn from the scope pictures. Three effects are not considered in the intepretation that is given. the first is that it is well known that degenration of red blood cells results in long sting like proteins being given off. These appear to be attached to the RBC...just like the pictures.

The next consideration is the heat from the dark field lamp... it is considerable and the result is thermal agitation of the fluid on the slide- the result is apparent movement of the observed inclusions.

The last factor that is not considered is the reality that at room temperature, certain protiens will stick together and precipitate into visible masses... which will appear to move when looked at with the high power lamp.

I have an open mind but would warn against jumping to conclusions without proper investigative dialog and review by open minded (and lyme infected) peers.

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Lymeorsomething
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I'd love to tool around with a 'scope myself but am skeptical about this guy's videos. If our blood were teeming with that many bugs, I don't know where we'd get the energy to modify scopes and to get out of bed for that matter.

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Eight Legs Bad
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Hi David

It wouldn't really be correct to say that Borrelia "eats tissue" or "eats blood". It parasitises the body by feeding on compounds in our body, including sterols which is unusual in bacteria, but it could get nutrients from blood or from a tissue cell.

As for the tick's hemolymph, some scientists believe it is in spiral form within the tick at certain times and in cell wall deficient (L-form) at other times.

I won't pretend to know much about the technical side of setting up a darkfield microscope, but I do know that Mark as an engineer is pretty knowledgeable about that side and I believe he has taken into account all stuff like Brownian motion, appropriate type of lamp etc..

I have witnessed live blood microscopy where you can see a microbe bound across the screen in an unmistakeably purposeful movement. No Brownian motion or coagulated lump of protein could behave that way.

I suggest that people have a look at the site of the Danish LLMD Dr Kroun.

http://lymerick.net/videomicroscopy.htm

The videos unfortunately take a long time to load but if you have the computer-power and the patience it is well worth the weight. Some of the borrelia filmed in chronic Lymies' blood has been bound with fluorescent antibody.

This is highly specific - no coagulated lump of protein or debris of degenerated red blood cells could bind to specific antibody.

Some of the videos there also show the remarkable snake-like movement of what Dr Kroun describes as "pearls on a string".

The modern film findings actually correlate very closely with what early darkfield borreliologists saw in the relapsing fever borrelia, ie the cystic and granular forms and the way they behave.

As for the person who says he can't believe a person could have so many spirochetes and still be able to move - in an infection, the numbers of infecting microbes are not the only factor determining how ill a person will be.

Whether the bacteria are active or dormant, whether they are producing toxins, how they affect the cells, where they are in the body, what kind of battle the immune system is waging - all these are factors too. Remember, people can co-exist with a population of microbes for decades (eg varicella zoster - chicken pox) - then suddenly an event like a drop in their immune system can bring the disease back to life, perhaps in a different form (shingles).

As for Mark's health, like a lot of chronic Lymies he has had periods of being severely ill, and also periods where he has been more healthy and able to work.

Elena

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Eight Legs Bad
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Also folks please remember that Miklossy has found spirochetes in the ***blood*** of Alzheimers patients as well as the brain tissue, and there have been a number of other findings published in the peer-reviewed literature.

Elena

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david1097
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I am very familiar with this topic and have posted in the past in this area.

I was not refering to the bacteria that "eats blood" but rather the tick. It has no method of consuming tissue, only blood and it is the only proven vector. The bacteria must therefore be in the blood of the animals that it bites.

I have seen thise videos. The main one showing movement is from a culture, not blood/ The second one uses blood but has some additive of undisclosed composition.

As far as marks descirption goes, I find the technical write up as it explains the scope, leds etc to be very good and it is a relatively easy issue to deplicate the design he has provided. Still it must be absolutely understood that the analysis of the observations are conjecture and I do not feel sufficient warnings are provided to readers as to the possibility of another possible intepretation.

A couple of simple tests can be done to eliminate the issues I have raised.


The first is simply to use a heated scope stage. This is common practice and in fact heated stages can be purchsed on ebay for less han $100 dollars. The stage can also be heated with a resistor, quite easily. This would eliminate the protein condensation issue I have raised.

What about verification the motion is not "brownian" motion but rather live bacteria moving,(which I believe is fluid convection, not brownian)?

This can simply be checked by exposing the slide to UV light and killing the bacteria. This can be done with an EPROM erasing lamp, a device readily avaialble to electrical enginners. If the motion stops after lamp exposure, the the motion being seen is due to the "bacteria" being alive. If the motion is still there then the things seen are simply not bacteria.


If the above two simple steps had been taken I would have a very different oppinion of the experiment results and interpretation that has been presented but without them I can only ask why not verify what is being proposed?

I have been around here since before mark has presented his findings (and know how he went about reaching his conclusions as they were discussed openly on the net). I have verified what he has seen under the scope, as have others like james. I don't disagree with what he has seen, rather I disagree with the interpretation of the observations.

As far as relapsing fever diagnosis using a microscope, as with virtually all other microscopic based diagnosis, stains are a central and key component to the diagnostic process. They are used in relaspsing fever diagnosis. Here is a good example of the process.

http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2593045

"live" blood analysis just leaves too many variables to deal with.

In the end, as stated, I have an open mind in this area. What I disagree with and what I am warning others about is that what has been presented cannot be taken as fact. This is important since this disease is so controversial any misrepresentation that is propagated, even if not deliberate, damages the cause and further discredits us as a whole group.

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oxygenbabe
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Thanks for your comments, eight legs and David.

I just want to note that the tick feeds for a while, and as it feeds on mammal's blood, which is a different temperature and different composition, this signals to the spirochete to begin migrating out of the tickgut and to begin, if I recall, expressing different proteins etc to ready itself for life in the blood of a very different host.

Seems to me the reverse could be true. As the tick injects various substances into our own blood, it might signal our spirochetes to move into our blood and to transfer themselves into the tick.

Both could be happening at once. Some mammals are already infected with the same or different strains. A tick might go away from its blood meal having transferred its own strain and acquired a new strain.

These bugs and hosts/vectors have evolved strategies over a great period of time. So both host and tick spirochetes would be sensitive to the chemicals released from each other, and would accordingly change their state/behavior to ready themselves for transmission.

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david1097
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That is an interesting concept and one that should be investigated. It would have to mean that the injected "chemical" is very potent but that is quite possible since ticks can cause tick paralysis in some people.

If the theory were true it could revelutionize diagnosis and maybe treatment. It might be easy to test, i'll give it some thought and mention it to some freinds of mine.

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Lymeorsomething
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quote:
Originally posted by Eight Legs Bad:
I have witnessed live blood microscopy where you can see a microbe bound across the screen in an unmistakeably purposeful movement. No Brownian motion or coagulated lump of protein could behave that way.

And that's another critique of these videos. I didn't recognize any purposeful movement.

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jamescase20
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What?

I have a natpath doctor who does live blood microscopy to dx lyme and I learned the skill myself at home with a phase contrast scope.

Lyme DOES swim in plasma...and lyme can be forced out of blood cells to see them. Granted they dont prefer to live in plasma but they have to move around somehow...

I can find lyme in just about any blood drop I pull easy....and its consistent too.

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Keebler
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-


One thing - of many - different from other bacteria is that Spirochetes do not require the blood stream to move about. They can literally shoot like a rocket through tissue. That is one way they can burrow deeper.


-

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david1097
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huh?

Sorry James, him, referes to the fellow that has he web page in the UK, not you.

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tcw
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quote:
The primers used by which lab/labs?
My thinking was that non-specific labs (ie. not Igenex) would use commercial kits - most of the docs that I could find on commercial kits indicated they were based on the flagellin gene. This is a (barely) educated guess, so please take it as one.

quote:
It has been known for years that OspA varies widely between species (it's one of the reasons the LymeRix vaccine was never marketed in Europe) and so would seem a poor choice.

Yeah, the vaccine was a strange thing - provoke an antigenic response to OspA which is variant. Maybe that was based on observation that the expression of Osp's varies based on the environment - isn't OspA highly expressed in the tick gut but not in human infections?

quote:
You seem to have a lot of knowledge about these matters - are you a microbiologist by any chance?
The very farthest thing from it - just a guy with a sick kid, an Internet account and some time on my hands.
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Lymeorsomething
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Believe me, guys. I want to believe (any X-Files fans out there?). However, the videos included on the above link were somewhat blurry and the spirochetal movement did not seem purposeful as you say.

Then again I know very little about 'chetes under the 'scope anyway. Still I like convincing evidence like everyone else. [Smile]

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Keebler
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According to some of the lectures in the past ILADS seminars, this sort of scope may not be strong enough - and much of the ILADS research does indicate that spirochetes are not easily seen in the blood.


The DVDs of past ILADS seminars are available through Lyme Disease Association -

www.lymediseaseassociation.org/VideoOrder.html


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Eight Legs Bad
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Trying to respond to several people at once here, so sorry if this post is a bit frgamented...

David - I think your points are valid. Although Mark did have confirmation of the borrelia in his blood via positive PCR result, that still does not prove 100% that the forms seen in the film of his blood are Borrelia. He should perhaps have used the phrase "possible borrelia", and maybe tried the techniques you suggested.

However, Dr Kroun and doctors working with her have used specific antibody to confirm findings of spirochete-like forms in the blood of chronic Lyme patients. And several researchers have found clear evidence of spirochetes in the blood , like the German study I mentioned before, done with ACA (a ***late manifestation of lyme***) blood. So we know borrelia is in the blood of at least some chronic patients.

And this is in harmony of what we know about relapsing fever borrelia. Those borrelia were often reported in the past to have had an "invisible phase" in the blood, in between the fever flares.

Nothing could be seen down the darkfield microscope, but the blood was infective to animals.

The borrelia were "invisible" because they were in the L-form.

Nenet, Keebler - I am not sure which paper on tick saliva you are talking about. Was it the one done by the Chinese Army?

Lymeorsomething - I have personally witnessed purposeful movement of an organism through the peripheral blood smear of a person on which a Lyme doctor was doing live darkfield microscopy. Quite a number of Lymies here in the UK have had this test done by LLMDs and some have watched it live on a screen.

You believe there is Ilads research proving Lyme usually isnt to be found in the blood - what papers are you referring to?

I think those Ilads doctors who have said that are not referring to any research of their own, but merely accepting older studies, many of which were done by our enemies.

It is tempting to believe that Lyme leaves Lyme the blood early, and permanently, in the illness and sequestrates itself in the brain, joints etc, never to return to the blood, hence avoiding exposure to our immune system and hence we have so many serongeative Lymies.

Lyme certainly does sequestrate in certain sites (it's been proven), and those sites certainly are immune-privileged as well as difficult for antibiotics to penetrate. However, that does not mean that Lyme does not ***also*** return to the circulation.

Lyme can propel its way through our tissue. However, I would not agree that it can move "like a rocket".

If I had Lyme sequestered only in my brain and my right ankle joint, but got bitten by a tick on my left shoulder, I imagine those borrelia would have a damn hard time making their way back to the tick on my shoulder, even if there was a chemical attractant in the tick's saliva, if they were not allowed to use the bloodstream.

Tcw I'm very sorry to hear about your child. I hope he/she is making some progress? You are right, ospA vaccine was supposed to work via antibodies killing Lyme in the tick, but that would rest on the assumption that every person in America who gets Lyme gets infected by the species Bb sensu stricto and nothing else, (and possibly it would require that every US Lyme only ever gets infected with a strain reasonably close to the strain used to prepare the vaccine), with no possibility of any other.

As we now know that Americans can get a Lyme-like illness from a borrelia that not only isnt sensu stricto, it *****isnt even from the burgdorferi group****, the concept would seem to be flawed, no?

But I think if we discuss ospA here we will get right off the topic. So I will start a separate thread on ospA, as I planned to talk about it anyway, especially regarding the bizarre work Dattwyler is doing with it at the moment. I hope some of you will join in the discussion.

Happy new year!


Elena

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Justice will be ours.

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Andromeda13
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There's a new paper out saying how good live blood smears are for rapid diagnosis:

The added value of peripheral blood cell
morphology in the diagnosis and management of
infectious diseasespart 1: basic concepts

M Prokocimer and I Potasman
Postgrad. Med. J. 2008;84;579-585
doi:10.1136/pgmj.2008.069609


ABSTRACT
As automated blood cell analysers and sophisticated
diagnostic technologies become widespread, requests for
peripheral blood smear (PBS) examination--for the
diagnosis of infectious diseases--diminish. Yet, PBS
examination can provide rapid and invaluable information
on infection--host susceptibility, aetiology, severity, and
systemic impact. Besides direct visualisation of certain
microorganisms (for example, Plasmodium, Ehrlichia), PBS
examination may detect characteristic footprints left by
various infections on the morphology of blood cells, thus
yielding the cytologic clues of the disease (for example,
Dohle bodies, haemophagocytosis).

Additionally, PBS
examination may disclose certain infection predisposing
conditions (for example, May-Hegglin anomaly, hyposplenism),
and several infection related haematological
and systemic complications. Combined with a careful
medical history and physical examination data, all this
information may yield a speedy diagnosis, a rationalised
diagnostic work-up, and timely initiation of treatment. The
intention of the following review is to highlight the value
of PBS, and recommend that PBS examination should be
fostered in the diagnostic work-up of infectious diseases

Some examples are given in the text

Mycobacterium avium Neutrophils, monocytes
(intracytoplasmic)/Wright, acid fast
Rod shaped structures Godwin et al81

Borreliosis Extracellular/Wright, Acridine orange Flagellar motile protozoan Cooper82

Ehrlichia Neutrophils (intracytoplasmic)/Giemsa Morula Heller et al83

Bartonella bacilliformis Surface and interior of RBC/NA Coccoid and filamentous
structures
Kapff et al42


Orientia tsutsugamushi Mononuclear cells
(intracytoplasmatic)/immunoalkaline
phosphatase and direct
immunofluorescent
Granules--focal and diffuse Walsh et al84

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Eight Legs Bad
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Thanks, Andromeda, it's a really interesting paper.

They talk about how borrelia (among other pathogens) can give rise to a terrible condition in which your white cells start to engulf your red cells within the bloodstream. There is a photo of it too.

Elena

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Justice will be ours.

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