posted
I've been looking at Dr Marie Kroun's site a lot recently, because she has lots of videos of blood taken from people with Lyme. The long spirochetes are seen swimming around and the bleb forms can be seen leaving the spiros.
On her site you can also look up the early history of research into spirochetes, and it's amazing to realise that over 100 years ago, scientists such as Leishman and Hindle and Balfour were meticulously drawing borrelia and treponema(syphilis) that they could easily see under the dark field microscope.
In an old text book from the 1950s there are drawings and photos which show that microbiologists can distinguish between the different types of spirochete just by eye.
In 1948 the electron microscope could show a coiled bunch of borrelia spirochetes inside a cyst and compare it with a similar arrangement that the syphilis spiro goes into.
There is a thing which puzzles me a great deal, and that is why Lyme is not being diagnosed just by looking down the dark field microscope.
That's how I've had my diagnosis, here in the UK, but to make sure, my blood was also sent to Bowen, and they confirmed that the L-form of Borrelia was in the blood.
Some friends have said that some of the ILADS doctors will not use Bowen, they prefer to have Western Blots as evidence. Surely the microscope method would make all the others redundant. Perhaps it would make too many lab workers redundant?
Admitted it's not a DNA positive proof that the patient has lyme, but if there are lots of spirochetes, then they have something nasty. Even ordinary Relapsing Fever is a very awful disease to have.
I am totally horrified by what I've seen in my own blood - the long bleb-filled spirochetes, like strings of pearls, most of them
very motile even in a thin smear of blood that's been left for up to 2 or 3 days,
plus the glowing tangled mass of the cell-wall deficient form that the Bowen labs can label with fluorescent antigen-binding chemical.
Some of the historical knowledge shows how much was known about spirochetes a very long time ago. In 1952, they knew that Borrelia was inside the brains of Multiple Sclerosis victims. They didn't know which strain of Borrelia, but they stained them with silver and ruled out syphilis.
One of the other things that is becoming obvious is that the culture medium doesn't seem to work that well, and I've seen some researchers adding blood to the BSK culture to keep the Bb growing well. BSK seems to work for Bb that has been found in the cerebrospinal fluid, but not for the spirochetes that were in human blood.
If we go back to 1928, the relapsing fever borrelia were, even then, known to change, in their virulence and symptoms..
Every time they passed the bug through a new host, be it a chipmunk, or a squirrel, the strain changed. One human strain from a relapsing fever victim was able to produce 6 different strains in squirrels.
Supposing a spirochete really is a long chain of blebs, with most of their DNA/RNA being the same as each other's, but some blebs are super blebs, with extra DNA These ones are able to adapt to live in different environments, sometimes tick blood, or human, or squirrel, and they either grow into new spirochetes, or even join up with other blebs to make a hybrid spirochete. Each different hybrid is a strain, such as valaisiana, or garinii.
Perhaps Borrelia have been messed about with so much through the years that they have formed new species and strains as a result of being passed from one host to another and adapting each time.
The main point though is: - why do we have to have such specific determination of which spirochete is in our blood?
Surely a person whose blood is swarming with these bugs may be acknowledged to be suffering? Could there not be a count done on the blood smears to see how many spirochetes there are in each field of view, under the dark field microscope?.
I have seen papers where animal blood is examined in this way to see the numbers of borrelia, before the scientists went on to culture and then do PCR. What is the reason for not doing this for humans??? Why do human borrelia not thrive in the BSK media?
I now know so many people who have had their blood examined under the dark field microscope. Some have more spirochetes than others, with the illest people having the greatest numbers. The spirochetes do not die so easily at all, even though the blood is drying up and there have been no nutrients added.
Healthy people do not have swarms of spirochetes in their blood, this is obvious. Or is it? If some doctors are correct, about 10% of us could be carrying the borrelia, but showing no symptoms. A bit like TB in fact. That doesn't mean those of us who are sick should be ignored.
I sometimes wish I had TB and not Bb, then I could access treatment.
There must be a reason for what's going on, because the knowledge by 1928 was huge, by 1948 it was being done at electron microscope level. Has someone deliberately kept it all secret?
I know that in 1928 ( a good year for microbiologists!) the first paper was published showing that genetic material could be exchanged between bacteria, and bacteria of different species Someone went to all the trouble of proving this all again in the early 1990s.
. Even though they didn't know the actual conformation of DNA in 1928, they knew a lot about how it behaved, but not enough to be in control of everything they were doing.
It's strange that in 1974, all of the microbiology labs had to stop work for a while to assess just how dangerous their work was, re safety in the labs, all across the world because it was so important.
Is anyone following this? Does anyone else know why the BSK media are so poor at growing Bb when the little bug is so hardy in blood? Leishman in 1911 took blood from African chicks to look at their spirochete infection. Even when he'd given the chicks an antibiotic (salvarsan) and he couldn't see the spiros in their normal blood, he did a needle biopsy into their livers and the spiros were still alive and kicking there.
Lots of questions - I hope someone can tell me if I'm barking up the wrong tree as we say in the UK. Best wishes, and thanks Betty, I hope I've followed your advice and put in enough spaces. Trouble is, it makes my post look even longer. Andromeda
Posts: 180 | From UK | Registered: Nov 2005
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Goody Tooshooz
Unregistered
posted
Thanks, Andromeda, it really is an amazing site as you said. Most of the videos seem to be at this URL:
I noticed that one of the scientists who made the videos had used monoclonal antibodies provided by Dr Alan Barbour. Barbour is no friend of Lyme patients, he is an arch-fan of Alan Steere. He's also the one who invented the BSK medium. (Barbour-Stoenner-Kelly medium).
Wouldnt it be ironic if Barbour had deliberately rigged the BSK medium so that it would only detect **some** Lyme disease strains, and not detect the more serious ones, the ones everybody has? After all, we know Bb is a very fastidious organism that is extremely difficult to culture. A pinch too much of this, a touch too little of that - and presumably, you inhibit growth.
It would be funny if Barbour's monoclonal antibodies have now helped to identify Lyme in patients who have had Lyme ruled out in the past on the basis of the CDC's diagnostic approach, the very criteria he promotes..
Goody
quote:Originally posted by Andromeda13: I've been looking at Dr Marie Kroun's site a lot recently, because she has lots of videos of blood taken from people with Lyme. The long spirochetes are seen swimming around and the bleb forms can be seen leaving the spiros.
On her site you can also look up the early history of research into spirochetes, and it's amazing to realise that over 100 years ago, scientists such as Leishman and Hindle and Balfour were meticulously drawing borrelia and treponema(syphilis) that they could easily see under the dark field microscope.
In an old text book from the 1950s there are drawings and photos which show that microbiologists can distinguish between the different types of spirochete just by eye.
In 1948 the electron microscope could show a coiled bunch of borrelia spirochetes inside a cyst and compare it with a similar arrangement that the syphilis spiro goes into.
There is a thing which puzzles me a great deal, and that is why Lyme is not being diagnosed just by looking down the dark field microscope.
That's how I've had my diagnosis, here in the UK, but to make sure, my blood was also sent to Bowen, and they confirmed that the L-form of Borrelia was in the blood.
Some friends have said that some of the ILADS doctors will not use Bowen, they prefer to have Western Blots as evidence. Surely the microscope method would make all the others redundant. Perhaps it would make too many lab workers redundant?
Admitted it's not a DNA positive proof that the patient has lyme, but if there are lots of spirochetes, then they have something nasty. Even ordinary Relapsing Fever is a very awful disease to have.
I am totally horrified by what I've seen in my own blood - the long bleb-filled spirochetes, like strings of pearls, most of them
very motile even in a thin smear of blood that's been left for up to 2 or 3 days,
plus the glowing tangled mass of the cell-wall deficient form that the Bowen labs can label with fluorescent antigen-binding chemical.
Some of the historical knowledge shows how much was known about spirochetes a very long time ago. In 1952, they knew that Borrelia was inside the brains of Multiple Sclerosis victims. They didn't know which strain of Borrelia, but they stained them with silver and ruled out syphilis.
One of the other things that is becoming obvious is that the culture medium doesn't seem to work that well, and I've seen some researchers adding blood to the BSK culture to keep the Bb growing well. BSK seems to work for Bb that has been found in the cerebrospinal fluid, but not for the spirochetes that were in human blood.
If we go back to 1928, the relapsing fever borrelia were, even then, known to change, in their virulence and symptoms..
Every time they passed the bug through a new host, be it a chipmunk, or a squirrel, the strain changed. One human strain from a relapsing fever victim was able to produce 6 different strains in squirrels.
Supposing a spirochete really is a long chain of blebs, with most of their DNA/RNA being the same as each other's, but some blebs are super blebs, with extra DNA These ones are able to adapt to live in different environments, sometimes tick blood, or human, or squirrel, and they either grow into new spirochetes, or even join up with other blebs to make a hybrid spirochete. Each different hybrid is a strain, such as valaisiana, or garinii.
Perhaps Borrelia have been messed about with so much through the years that they have formed new species and strains as a result of being passed from one host to another and adapting each time.
The main point though is: - why do we have to have such specific determination of which spirochete is in our blood?
Surely a person whose blood is swarming with these bugs may be acknowledged to be suffering? Could there not be a count done on the blood smears to see how many spirochetes there are in each field of view, under the dark field microscope?.
I have seen papers where animal blood is examined in this way to see the numbers of borrelia, before the scientists went on to culture and then do PCR. What is the reason for not doing this for humans??? Why do human borrelia not thrive in the BSK media?
I now know so many people who have had their blood examined under the dark field microscope. Some have more spirochetes than others, with the illest people having the greatest numbers. The spirochetes do not die so easily at all, even though the blood is drying up and there have been no nutrients added.
Healthy people do not have swarms of spirochetes in their blood, this is obvious. Or is it? If some doctors are correct, about 10% of us could be carrying the borrelia, but showing no symptoms. A bit like TB in fact. That doesn't mean those of us who are sick should be ignored.
I sometimes wish I had TB and not Bb, then I could access treatment.
There must be a reason for what's going on, because the knowledge by 1928 was huge, by 1948 it was being done at electron microscope level. Has someone deliberately kept it all secret?
I know that in 1928 ( a good year for microbiologists!) the first paper was published showing that genetic material could be exchanged between bacteria, and bacteria of different species Someone went to all the trouble of proving this all again in the early 1990s.
. Even though they didn't know the actual conformation of DNA in 1928, they knew a lot about how it behaved, but not enough to be in control of everything they were doing.
It's strange that in 1974, all of the microbiology labs had to stop work for a while to assess just how dangerous their work was, re safety in the labs, all across the world because it was so important.
Is anyone following this? Does anyone else know why the BSK media are so poor at growing Bb when the little bug is so hardy in blood? Leishman in 1911 took blood from African chicks to look at their spirochete infection. Even when he'd given the chicks an antibiotic (salvarsan) and he couldn't see the spiros in their normal blood, he did a needle biopsy into their livers and the spiros were still alive and kicking there.
Lots of questions - I hope someone can tell me if I'm barking up the wrong tree as we say in the UK. Best wishes, and thanks Betty, I hope I've followed your advice and put in enough spaces. Trouble is, it makes my post look even longer. Andromeda
posted
Andromeda, I agree with what you have said 100%.
I was diagnosed in the UK with my doctor using Darkfield Microscopy.
He said he saw the little critters swimming everywhere, I'm quite glad I didn't see it.
I will be having the Bowen test to confirm, but I have been told everyone who has been told they have spirochetes in their blood by our doctor gets a positive Bowen test.
I think it should be used more often, instead of messing around with unreliable ELISA's and Western Blot's.
I believe in the Bowen test 100%, the high positive stats are just showing how common this is...
Posts: 263 | From UK | Registered: Mar 2006
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posted
Hey goody, you can reply without quoting the whole previous message. Can click on the little pencil to edit. You a newby? Welcome. Tell us your story sometime.
My comment on this post-- Modern lab medicine has a dangerous side effect that is unknown to these scientists. It rots their brains and sucks out any common sense they had in the beginning. Everything new is wonderful, everything old is worthless.
Posts: 8430 | From Not available | Registered: Oct 2000
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Does anyone know of the reason why the doctors and microbiologists don't look down the dark field microscope for borreliae?
Except for their own academic experimental purposes when they actually do need to find the spiros and then culture them!! BW, Andromeda
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Goody Tooshooz
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posted
There is talk that maybe the reason everyone thinks it's not worthwhile looking directly at the blood under darkfield microscope, is because in the past when doctors did that to chronic Lymies' blood, they cultured it in BSK medium first, and then couldnt find anything. Maybe the BSK medium restricted growth of the strains we all have.
Maybe it's only good for growing the "easily cured in 3 weeks strain" that no one seems to have! (I know a lot of Lymies, but none of them were "easily cured in 3 weeks"!)
Have a look at this:
"It has been demonstrated previously that motile Borrelia burgdorferi cells transform into non-motile cyst-forms when incubated for several weeks in BSKII (a complex medium) lacking rabbit serum.
B. burgdorferi cells cannot synthesize fatty acids de novo and serum is thought to provide a source of fatty acids and lipids. When B. burgdorferi cells were serum-starved in defined RPMI medium, ~90% of the cells formed spherical cysts within 48 h.
Cyst formation was inhibited by tetracycline. Cyst opening and recovery of vegetative cells was rapidly induced by the addition of either BSKII or rabbit serum.
The percentage of viable cells recovered from cysts ranged from 2�9% to 52�5%. Viability was inversely proportional to cyst age.
Protein synthesis by B. burgdorferi during serum starvation was examined by labelling cells with Tran35S-Label and analysing the labelled proteins by two-dimensional gel electrophoresis and fluorography. The synthesis of over 20 proteins was induced during serum starvation.
Western blots of proteins from vegetative cells and cysts probed with sera from either B. burgdorferi-infected humans or monkeys revealed that several cyst proteins were antigenic.
These data suggest that cells of B. burgdorferi, although possessing a small genome and extremely limited biosynthetic capabilities, rapidly respond to conditions of serum starvation by inducing changes in protein synthesis and cell morphology.
This study may help explain how cells of B. burgdorferi can survive periods of nutrient deprivation in different hosts and host tissues."
That's from a guy called Alban, it's on Pubmed.
If putting it in a medium not as good as BSK makes it go cystic, maybe putting another strain in Bb means you get it going cystic (and therefore not showing up on the microscopy) , until you out it in a medium that is BETTER than BSK.
But these doctors in England and Denmark and other places are seeing it in the blood under darkfield microscope without growing it in ANY medium at all first. They just look at a drop of blood and see it.
Maybe we should all get ourselves a microscope, set up a darkfield (there are textbooks on the internet telling you how to do this, it's not that difficult), and have a look and see what's in our blood.
posted
I am not sure if you can see B. burgdorferi spirochetes with dark-field microscopy in the blood of most Lyme patients.
Because the Ignex PCR tests usaully are negative.
The PCR is detecting B. burgdorferi directly like microscopy but much more sensitive.
If PCR cannot detect B. burgdorferi in the blood, the success by microscopy is very very low or impossible.
Posts: 77 | From USA | Registered: Feb 2006
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Goody Tooshooz
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posted
Bluesky, that's not correct.
You see the PCR test is only as good as the DNA primers it's based on.
Say you had a different strain of Borrelia, which was very different to the ones that the primers we have available can detect, but it still caused Lyme. (Like the b. lonestari in the south is very different to B. burgdorferi)
It wouldnt show on PCR, but you would still be able to see it under darkfield microscope, as long as you didn't try growing it in BSK first. Theoretically anyway.
I guess we wont know for sure till everybody tries it, or at least gets their doctor to try it, if they cant lay their hands on a micrscope themselves.
All the available primers used in Lyme pcr's today must be based on the original strain that Alan Barbour cultured, back when Burgdorfer discovered the Lyme bacterium.
Suppose his (Barbour's) prototype wasn't really like the strains everybody has, but only like the Steere rosy view of Lyme disease that almost no one gets? I mean, who trusts Alan Barbour, after all the crap he said about Lyme being hypochondria etc..
Lou, what do you mean about scientists think everything old is crap? I couldn't tell what you were trying to say.
quote:Originally posted by blueskyfaith: I am not sure if you can see B. burgdorferi spirochetes with dark-field microscopy in the blood of most Lyme patients.
Because the Ignex PCR tests usaully are negative.
The PCR is detecting B. burgdorferi directly like microscopy but much more sensitive.
If PCR cannot detect B. burgdorferi in the blood, the success by microscopy is very very low or impossible.
Ohh, you would be suprised. I had 1 drop (yes 1 drop) of blood taken and my doctor could see them swimming everywhere. If you see a spirochete, you have a spirochete infection. If PCR is more sensitive, why would everyone be complaining about the high positive results from Bowen and other doctors who use microscopy??? I never see people complain about high-positive PCR statistics.
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Goody Tooshooz
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posted
Well look at this from Willy Burgdorfer.
How can engorgement on infected hosts result in 100% infected ticks, if the borrelia are not present in the blood of the host when bitten by the tick, or are only present in tiny amounts?
Then you get Wormser saying you can detect it in the blood of early Lyme, but you need to add lots and lots of blood to the culture medium (9 ml, which is a lot). Well maybe its because you're adding blood, containing nutrients the bug needs, which are missing in the BSK culture medium?
Below I copied the Burgdorfer article.
Goody
"This relatively large Borrelia [Borrelia burgdorferi] is not readily detectable in blood smears or thick drops of Lyme disease patients and susceptible host animals, yet engorgement on infected hosts results in up to 100% infected ticks....
RML [NIH's Rocky Mountain Lab] scientists Dave Dorward and Claude Garon using silver staining, transmission and scanning electron microscopy investigated the nature of naturally elaborated membrane blebs on the surface of cultured B. burgdorferi or free in the medium, and found both linear and circular DNA (Fig.13)...
These most recent findings [of RML researchers and others] do confirm the development of membrane-derived cysts, blebs, spherules, vesicles and the potential transformation to motile, helical spirochetes...as a "survival mechanism" of spirochetes to overcome or escape unfavorable conditions."
[12th International Conference on Lyme Disease and Other Spirochetal and Tick-Borne Disorders., April 8-9, 1999 Keynote Address - The Complexity of Vector-borne Spirochetes. Burgdorfer, W.
Willy Burgdorfer, Ph.D., of the National Institutes of Health, is the discoverer of Borrelia burgdorferi.]
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Of course, I assumed that the PCR technique was working including that the primers were good to recognize the DNA.
There are at least two spirochetes in Borrelia genera that their complete genomic sequences have been sequenced.
Based on these sequences, one should be able to design primers that will detect the two species, hence their every strains.
I assumed that Igenex was using at least species-specific if not genera-specific primers, and should have considered the strain problems you mentioned. Otherwise, I would doubt that the Igenex scientists know the basics of molecular biology.
Posts: 77 | From USA | Registered: Feb 2006
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I agree with you that microscopy is probably the best way to detect germs in the blood as long as the germs present in a detectable amount, such as in your case.
quote:If you see a spirochete, you have a spirochete infection.
However, most LD patients are not as lucky as you, they simply don't have spirochetes in the blood enough to be detected by microscopy.
quote: If PCR is more sensitive, why would everyone be complaining about the high positive results from Bowen and other doctors who use microscopy??? I never see people complain about high-positive PCR statistics.
Because, my guess is that PCR is very specific, thus the false posivtive is very low.
Posts: 77 | From USA | Registered: Feb 2006
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May I ask how many Lymies you know of who have had someone look for spirochetes in their blood? And what percentage were negative?
If the situation in the States is the same as the UK, then my guess is that no one normally has their blood examined by dark field microscopy, and that nearly 100% of people who are suspected lyme patients just have an ELISA.
It's also extremely rare for anyone to get as far as having a PCR in the UK, as hardly anyone knows of it, not even the physicians. A few privately-paying patients might have this arranged.
In the UK I have spoken to people with a known tick bite and many symptoms, who have actually been told by their doctors that they cannot have a Lyme test, because only one or two people a year get Lyme in the UK!
One woman was told she had "Scarletina" and had had it several times (!!) while another was told it was just "Ringworm". Still another was told that she had a lot of different illnesses, and the lyme wasn't worth testing for, since it only lasts for a few weeks. All these people had multiple symptoms, including thyroid hormone deficiency in one case.
Such is the ignorance of the UK doctors. Perhaps the microbiology labs are just as outdated, or badly directed.
But I would be very interested to find out, Blue Sky, if some of your friends or acquaintances had experience of the dark field microscopy, because I really want to get to the truth of this.
Perhaps it does depend on the vigilance and experience of the person looking down the microscope, or how sick the patient is, or if they have been taking antibiotics.
Best wishes, Andromeda
Posts: 180 | From UK | Registered: Nov 2005
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Thanks and I appreciate your determination on finding out the truth.
You might be right that microscopy might be superior to ELISA test, which basically is usless.
I don't have an answer for your question. My statment was based on what I read and the Igenex results people posted on the web.
However, I did look several blood smears under phase microscope (similar to dark field), and I didn't see a single spirochete.
While the blood was Lyme CDC positive by Western blots.
Anyway, I do encourage you or other people to look blood under microscope to check for infections, not just for Lyme sprochates, but for other co-infections like Babes., Bart. etc.
Because it's very simple: you just need a microscope probably 20X100 power.
As far as I know, so far, the most reliable test for Bbs is Western blots from Igenex.
I hope this would be of any help.
Good luck.
Posts: 77 | From USA | Registered: Feb 2006
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I've now followed your link to the topic "I want my own microscope". Wish I'd seen all this before, it means I should look at Lymenet more often.
Wow, so many enterprising people all being able to do their own microscopy. I feel in awe somewhat, but so pleased that people can do it for themselves.
It definitely seems spooky that the doctors don't seem to bother much with darkfield, except a few enlightened ones. All I know is that the ones in my blood are long, bent into regular waves along their length, and full of blebs. But the doctors who will recognise the tests like this in the UK are very rare.
I don't know how long our NHS and HPA can ignore the truth, and for what purpose they deliberately want to leave Lyme patients without treatment.
Do you know anything about the Tuskegee US experiment, when they left people, most of whom were black Americans, with no treatment for their syphilis, to see what would happen?
I must google for this, as I've never looked for the details before, to see whether the results of this experiment might still be affecting us now, in that they believed that no treatment was just as good as giving them penicillin.
The health agencies might believe that the expense isn't worth a few hundred thousand people living tortured lives.
How do they think the whole population will end up if they allow the Lyme plague to continue? I just don't know how they sleep at night with this on their consciences.
Best wishes and take care Blue Sky, Andromeda
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Goody Tooshooz
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posted
There are cases in the medical literature of borrelia being present but missed even on genus-specific PCR.
Borrelia lonestari does not show on a Bb pcr, and does not grow in BSK medium, but it causes a disease like Lyme disease. If this borrelia species, which is not part of the burgdorferi complex can cause Lyme symptoms, then maybe other (non-burgdorferi) species can too?
Maybe they can make a person sick as a dog, but evade most PCR's,and wont grow in BSK, just like Lonestari, but when you look down a microscope at an ordinary drop of blood you see them?
Btw, the smears you examined with phase-contrast, did anyone check them with darkfield?
Thanks. Goody
quote:Originally posted by blueskyfaith:
I assumed that Igenex was using at least species-specific if not genera-specific primers, and should have considered the strain problems you mentioned.
posted
I will be getting a darkfield scope in about 3 weeks hopefully, so I will let you know how I get on.
Posts: 263 | From UK | Registered: Mar 2006
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posted
This is how another spirochetal infection was studied for decades.
How long will it be before we get some truth out in the media about how our illness with the borrelia spirochete is being ignored, denied, left untreated and the whole topic kept quiet in all mainstream debate? The word Lyme is still unknown here in Britain, which makes our government guilty of criminal neglect.
How many doctors now are prepared to acknowledge what's going on? The world goes wrong when good people do nothing. Those who know anything about all of this, please examine your consciences and remember that the truth always comes out in the end.
The Tuskegee Syphilis Experiment The United States government did something that was wrong--deeply, profoundly, morally wrong. It was an outrage to our commitment to integrity and equality for all our citizens. . . . clearly racist. --President Clinton's apology for the Tuskegee Syphilis Experiment to the eight remaining survivors, May 16, 1997 For forty years between 1932 and 1972, the U.S. Public Health Service (PHS) conducted an experiment on 399 black men in the late stages of syphilis. These men, for the most part illiterate sharecroppers from one of the poorest counties in Alabama, were never told what disease they were suffering from or of its seriousness. Informed that they were being treated for ``bad blood,''1 their doctors had no intention of curing them of syphilis at all. The data for the experiment was to be collected from autopsies of the men, and they were thus deliberately left to degenerate under the ravages of tertiary syphilis--which can include tumors, heart disease, paralysis, blindness, insanity, and death. ``As I see it,'' one of the doctors involved explained, ``we have no further interest in these patients until they die.''
Using Human Beings as Laboratory Animals The true nature of the experiment had to be kept from the subjects to ensure their cooperation. The sharecroppers' grossly disadvantaged lot in life made them easy to manipulate. Pleased at the prospect of free medical care--almost none of them had ever seen a doctor before--these unsophisticated and trusting men became the pawns in what James Jones, author of the excellent history on the subject, Bad Blood, identified as ``the longest nontherapeutic experiment on human beings in medical history.''
The study was meant to discover how syphilis affected blacks as opposed to whites--the theory being that whites experienced more neurological complications from syphilis whereas blacks were more susceptible to cardiovascular damage. How this knowledge would have changed clinical treatment of syphilis is uncertain. Although the PHS touted the study as one of great scientific merit, from the outset its actual benefits were hazy. It took almost forty years before someone involved in the study took a hard and honest look at the end results, reporting that ``nothing learned will prevent, find, or cure a single case of infectious syphilis or bring us closer to our basic mission of controlling venereal disease in the United States.'' When the experiment was brought to the attention of the media in 1972, news anchor Harry Reasoner described it as an experiment that ``used human beings as laboratory animals in a long and inefficient study of how long it takes syphilis to kill someone.''
A Heavy Price in the Name of Bad Science By the end of the experiment, 28 of the men had died directly of syphilis, 100 were dead of related complications, 40 of their wives had been infected, and 19 of their children had been born with congenital syphilis. How had these men been induced to endure a fatal disease in the name of science? To persuade the community to support the experiment, one of the original doctors admitted it ``was necessary to carry on this study under the guise of a demonstration and provide treatment.'' At first, the men were prescribed the syphilis remedies of the day--bismuth, neoarsphenamine, and mercury--but in such small amounts that only 3 percent showed any improvement. These token doses of medicine were good public relations and did not interfere with the true aims of the study. Eventually, all syphilis treatment was replaced with ``pink medicine''--aspirin. To ensure that the men would show up for a painful and potentially dangerous spinal tap, the PHS doctors misled them with a letter full of promotional hype: ``Last Chance for Special Free Treatment.'' The fact that autopsies would eventually be required was also concealed. As a doctor explained, ``If the colored population becomes aware that accepting free hospital care means a post-mortem, every darky will leave Macon County...'' Even the Surgeon General of the United States participated in enticing the men to remain in the experiment, sending them certificates of appreciation after 25 years in the study.
Following Doctors' Orders It takes little imagination to ascribe racist attitudes to the white government officials who ran the experiment, but what can one make of the numerous African Americans who collaborated with them? The experiment's name comes from the Tuskegee Institute, the black university founded by Booker T. Washington. Its affiliated hospital lent the PHS its medical facilities for the study, and other predominantly black institutions as well as local black doctors also participated. A black nurse, Eunice Rivers, was a central figure in the experiment for most of its forty years. The promise of recognition by a prestigious government agency may have obscured the troubling aspects of the study for some. A Tuskegee doctor, for example, praised ``the educational advantages offered our interns and nurses as well as the added standing it will give the hospital.'' Nurse Rivers explained her role as one of passive obedience: ``we were taught that we never diagnosed, we never prescribed; we followed the doctor's instructions!'' It is clear that the men in the experiment trusted her and that she sincerely cared about their well-being, but her unquestioning submission to authority eclipsed her moral judgment. Even after the experiment was exposed to public scrutiny, she genuinely felt nothing ethical had been amiss.
One of the most chilling aspects of the experiment was how zealously the PHS kept these men from receiving treatment. When several nationwide campaigns to eradicate venereal disease came to Macon County, the men were prevented from participating. Even when penicillin was discovered in the 1940s--the first real cure for syphilis--the Tuskegee men were deliberately denied the medication. During World War II, 250 of the men registered for the draft and were consequently ordered to get treatment for syphilis, only to have the PHS exempt them. Pleased at their success, the PHS representative announced: ``So far, we are keeping the known positive patients from getting treatment.'' The experiment continued in spite of the Henderson Act (1943), a public health law requiring testing and treatment for venereal disease, and in spite of the World Health Organization's Declaration of Helsinki (1964), which specified that ``informed consent'' was needed for experiment involving human beings.
Blowing the Whistle The story finally broke in the Washington Star on July 25, 1972, in an article by Jean Heller of the Associated Press. Her source was Peter Buxtun, a former PHS venereal disease interviewer and one of the few whistle blowers over the years. The PHS, however, remained unrepentant, claiming the men had been ``volunteers'' and ``were always happy to see the doctors,'' and an Alabama state health officer who had been involved claimed ``somebody is trying to make a mountain out of a molehill.''
Under the glare of publicity, the government ended their experiment, and for the first time provided the men with effective medical treatment for syphilis. Fred Gray, a lawyer who had previously defended Rosa Parks and Martin Luther King, filed a class action suit that provided a $10 million out-of-court settlement for the men and their families. Gray, however, named only whites and white organizations in the suit, portraying Tuskegee as a black and white case when it was in fact more complex than that--black doctors and institutions had been involved from beginning to end.
The PHS did not accept the media's comparison of Tuskegee with the appalling experiments performed by Nazi doctors on their Jewish victims during World War II. Yet in addition to the medical and racist parallels, the PHS offered the same morally bankrupt defense offered at the Nuremberg trials: they claimed they were just carrying out orders, mere cogs in the wheel of the PHS bureaucracy, exempt from personal responsibility.
The study's other justification--for the greater good of science--is equally spurious. Scientific protocol had been shoddy from the start. Since the men had in fact received some medication for syphilis in the beginning of the study, however inadequate, it thereby corrupted the outcome of a study of ``untreated syphilis.''
In 1990, a survey found that 10 percent of African Americans believed that the U.S. government created AIDS as a plot to exterminate blacks, and another 20 percent could not rule out the possibility that this might be true. As preposterous and paranoid as this may sound, at one time the Tuskegee experiment must have seemed equally farfetched. Who could imagine the government, all the way up to the Surgeon General of the United States, deliberately allowing a group of its citizens to die from a terrible disease for the sake of an ill-conceived experiment? In light of this and many other shameful episodes in our history, African Americans' widespread mistrust of the government and white society in general should not be a surprise to anyone. --BB
1. All quotations in the article are from Bad Blood: The Tuskegee Syphilis Experiment, James H. Jones, expanded edition (New York: Free Press, 1993).
Information Please� Database, � 2006 Pearson Education, Inc. All rights reserved.
Posts: 180 | From UK | Registered: Nov 2005
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Dave6002
Frequent Contributor (1K+ posts)
Member # 9064
posted
Hi, Andromeda and Goody,
You guys are right about seeing Bbs under darkfield microscope.
Please see my new post. I saw the enemies in the blood today.
I couldn't thank you enough for bring up this question.
Otherwise I would be still in dark: thinking that Bbs are in deep tissues not in blood.
posted
All of this makes me wonder just what the growth media do to the spirochetes.
Does anyone know a good place to see the history of who developed the BSK medium, and why they always seem to be modifying it with this and that?
I've seen a few papers from different labs and at different times, and it's really difficult to follow what was developed and when. Looks like more research needed here.
Well done Dave with the microscope, looking forward to seeing them if you are able to do the pictures, Best wishes, Andromeda
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posted
Hi Goody 2 shoes, I've realised it's your posts that have made me think about the media, and having a Lyme brain I forgot - so thanks for stimulating a very relavant thought process!
I did see so many references to media that I didn't know where to start, but I came across this one where they say that the culturing alters things in an important way:
JOURNAL OF CLINICAL MICROBIOLOGY, Sept. 1997, p. 2359-2364 Vol. 35, No. 9 Culturing Selects for Specific Genotypes of Borrelia burgdorferi in an Enzootic Cycle in Colorado DOUGLAS E. NORRIS,1� BARBARA J. B. JOHNSON,2 JOSEPH PIESMAN,2 GARY O. MAUPIN,2 JESSICA L. CLARK,1 AND WILLIAM C. BLACK IV1* Department of Microbiology, Colorado State University, Ft. Collins, Colorado 80523,1 and Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado 805222
Received 7 March 1997/Returned for modification 13 May 1997/Accepted 11 June 1997
In Colorado, Borrelia burgdorferi sensu stricto, the etiologic agent of Lyme disease, is maintained in an enzootic cycle between Ixodes spinipalpis ticks and Neotoma mexicana rats (27). The frequencies of flagellin (fla), 66-kDa protein (p66), and outer surface protein A (ospA) alleles were examined in 71 B. burgdorferi isolates from samples from Colorado. Approximately two-thirds of these samples were isolates from I. spinipalpis ticks that had been cultured in BSK-H medium prior to DNA extraction. The remaining samples were from total DNA extracted directly from infected I. spinipalpis ticks. A portion of each gene was amplified by PCR and screened for genetic variability by single-strand conformation polymorphism (SSCP) analysis. We identified three alleles in the fla gene, seven in the p66 gene, and seven in the ospA gene. Sequencing verified that the amplified products originated from B. burgdorferi template DNA and indicated 100% sensitivity and specificity of the SSCP analysis. The frequencies of the p66 and ospA alleles were significantly different between cultured and uncultured spirochetes. The number of three-locus genotypes and the genetic diversity of alleles at all loci were consistently lower in cultured spirochetes, suggesting that culturing of B. burgdorferi in BSK-H medium may select for specific genotypes.
Complicated stuff, but interesting. Best wishes Goody, Andromeda
Posts: 180 | From UK | Registered: Nov 2005
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Dave6002
Frequent Contributor (1K+ posts)
Member # 9064
posted
Hi,Andromeda,
Bbs are parasites so the media could be very tricky: one thing, thay cannot use even simple suger like glucose, cuz they don't have enzymes to use glucose to produce energy ATP. So adding ATP to the media might not be a bad idea. also the oxygen, they don't need much. Temperature, they need 34C.
Could we Lymies go together and set up a lab and work to save our and others life?
Thanks and take care
Posts: 1078 | From Fairland | Registered: Apr 2006
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posted
Hi Dave 6002, Just wanted to say that if I could get a bit more well, I would work as an unpaid volunteer staring down microscopes and recording the relevant frames for a doctor to analyse. It wouldn't be boring, because it's so worthwhile.
One day if you guys get this together I would do all I could to help in any way. I wish this damned disease had not laid me so low, but I am half way to getting the right treatment, so I'll be optimistic.
Good luck with all your efforts. Andromeda
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posted
I had dark field microscopy done after being on months of combo abx, and the MD pathologist doing it managed to tape a video of a spirochete gaily swimming along in one drop of my blood. However, she said we could not positively identify it as a Bb spirochete--some spirochetes are less problematic than others, such as those commonly found in the mouth that can cause gum disease. That's why culturing is important, and Bb are notoriously difficult to culture. Nevertheless, my infectious disease doc was impressed enough to show the video to his students and ID dept. Many of his thoughts on lyme changed 180' after dealing with my case. (He was originally in the Dr. Alan Steere camp, but fortunately was more open-minded than most.)
Posts: 193 | From Virginia | Registered: Oct 2002
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posted
Some researchers say Dark field was better than PCR and culture in this series of references:
(This is copied straight from a canlyme post about the same topic, don't suppose they will mind).
1: Arch Pathol Lab Med. 1983 Jul;107(7):384-6.
Detection of Borrelia in acridine orange-stained blood smears by fluorescence microscopy.
Sciotto CG, Lauer BA, White WL, Istre GR.
Tick-borne borreliosis (relapsing fever) can be an important, unsuspected cause of febrile illness. The diagnosis is generally made by identifying Borrelia spirochetes in stained peripheral blood smears. Since Borrelia may be difficult to detect with Romanowsky stains, an alternative method, using acridine orange (AO), was used to screen blood smears. Duplicate blood smears of seven patients were examined with the AO technique and Romanowsky stains. In all seven cases spirochetes were easily identified with the AO-stained smears compared with only five cases with Romanowsky stains. In a double-blind laboratory experiment, six of ten duplicate smears from a single patient with mild spirochetemia were positive by AO, whereas only two of ten were positive by Romanowsky stain. We concluded that the AO stain is simple, rapid and more sensitive than Romanowsky methods for detecting cases of low-level spirochetemia.
Wien Klin Wochenschr (2002) 114/13-14: 601-605
Limitation of serological testing for Lyme borreliosis: Evaluation of ELISA and Western blot in comparison with PCR and culture methods
Stanislawa Tylewska-Wierzbanowska and Tomasz Chmielewski
Summary. The aim of the study was to evaluate a one-step procedure using an ELISA test of high specificity and a two-step procedure using immunoblot as a confirmation test, and to compare the results of serological testing with detection of bacterial DNA and living spirochetes.
Sera, synovial (SF) and cerebro-spinal fluids (CSF) were obtained from 90 patients with clinical symptoms of Lyme borreliosis. Serum samples were tested with recombinant ELISA and Western blot assay. Citrated blood, cerebrospinal and synovial fluids samples were cultured in cell line and tested by PCR to detect spirochetes.
No correlation was found between levels of specific B. burgdorferi antibodies detected with a recombinant antigen ELISA and the number of protein fractions developed with these antibodies by immunoblot. Moreover, Lyme borreliosis patients who have live spirochetes in body fluids have low or negative levels of borrelial antibodies in their sera. This indicates that an efficient diagnosis of Lyme borreliosis has to be based on a combination of various techniques such as serology, PCR and culture, not solely on serology.
APPLIED MICROBIOLOGY, Oct. 1974, p. 540-543 Vol. 28, No. 4
Biology of Borrelia hermsii in Kelly Medium HERBERT G. STOENNER
More than 800 Borellia hermsii in mouse plasma were required for establishment of growth in an artificial medium (Kelly), but only a single organism of a fully adapted strain (25th subculture) was required for a successful subculture. As judged by generation time, maximal concentration in culture, and length and motility of the organism, the process of adaptation extended through at least 11 subcultures. Because the organisms regularly died shortly after the logarithmic growth phase, transfers at 7- to 10-day intervals were required to maintain continuous cultures.
Spirochetes in the medium and citrated mouse plasma were counted under a cover slip with a dark-field microscope. As delivered from a 0.005-ml calibrated capillary pipette (Kimble microcapillary pipette), 0.0025 ml of medium was deposited on a slide, surrounded by a ring of a mixture containing petrolatum and plasticine (2:1, wt/wt), and covered with a cover slip, and sufficient pressure was applied to extend the drop to a diameter of about 8 mm. Counts were made of dilutions containing 3 to 12 organisms per 0.0025 ml; the entire area was searched for borreliae at a magnification of x 125. All counts were made in duplicate and averaged. It should be emphasized that cover slips, slides, and capillary tubes must be cleaned thoroughly. A method was devised for culturing a single spirochete in sterile capillary tubes (90 mm long, 0.4 mm inner diameter). In this method, the number of spirochetes in vigorously growing cultures (5 to 7 days old) or mouse plasma was first established by direct count.
Parazitologiia. 2001 Jan-Feb; 35(1): 3-8.
Comparative evaluation of the efficacy of Borrelia indication in ixodes ticks (Ixodidae) using dark field microscopy and polymerise chain reaction (PCR)
Indication of Borrelia (B. burgdorferi sensu lato) in 205 adult unfed I. persulcatus ticks from a natural focus was carried out simultaneously by methods of PCR and dark-field microscopy of vital preparations. PCR method revealed Borrelia prevalence in a considerable number of ticks, in which Borrelia were not found by microscopy of 250 microscopic fields in a preparation from each individual tick. At the same time, PCR method didn't give positive results for approximately 8% of ticks, which contained rather high concentration of Borrelia (more than 10 per 100 microscopic fields). In general, PCR method doesn't have advantages in comparison with a microscopy of vital preparations for study of the Borrelia prevalence in ticks.
Journal of Clinical Microbiology, May 1999, p. 1361-1365, Vol. 37, No. 5
Prevalence of Borrelia burgdorferi in Ixodes ricinus Ticks in Urban Recreational Areas of Helsinki
Juha Junttila,1 Miikka Peltomaa,2,* Hanna Soini,3 Merja Marjam�ki,3 and Matti K. Viljanen3
Lyme borreliosis, an infection caused by the tick-borne spirochete Borrelia burgdorferi, is a major health problem for populations in areas of endemicity in the Northern Hemisphere. In the present study we assessed the density of ticks and the prevalence of B. burgdorferi sensu lato among ticks in popular urban recreational areas of Helsinki, Finland. Altogether 1,688 Ixodes ricinus ticks were collected from five areas located within 5 km of the downtown section of Helsinki, and 726 of them (303 nymphs, 189 females, and 234 males) were randomly chosen for laboratory analysis. The midguts of the ticks were divided into three pieces, one for dark-field microscopy, one for cultivation in BSK-II medium, and one for PCR analysis. Ticks were found in all the study areas; their densities varied from 1 to 36 per 100 m along which a cloth was dragged. The rate of tick infection with B. burgdorferi sensu lato varied from 19 to 55%, with the average being 32%. Borellia afzelii was the most predominant genospecies in all the areas, and no B. burgdorferi sensu stricto isolates were detected. Only two ticks were concurrently infected with both B. afzelii and Borrelia garinii. Dark-field microscopy gave more positive results for B. burgdorferi than did cultivation or PCR analysis. However, the agreement between all three methods was fairly good. We conclude that Lyme borreliosis can be contracted even in urban environments not populated with large mammals like deer or elk. The disease should be taken into account in the differential diagnosis of certain symptoms of patients from these areas, and the use of measures to improve the awareness of the general population and health care officials of the risk of contracting the disease is warranted. DF microscopy. One of the three parts of the tick midgut was placed on a microscope glass in a drop of BSK-II medium and was covered with a cover glass. The number of spirochetes in the midgut sample was estimated by DF microscopy (Laborlux D; Leitz, N�rnberg, Germany) by examining 100 fields at a magnification of �400. Typical movement, morphology, and size were used as the identification criteria for the borreliae.
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