‘Granulated Cellular Structures and other abnormal microscopy findings in blood from chronically ill patients’

When I started looking in a microscope and I had a background from pathology, I looked at cells, so I had a little basis for it. At the last Conference I gave lecture on history and I omitted some important works in history that I will talk about today. The main works that I want to speak about are:

In 1911: A scientist called Balfour – his work describing (these illnesses) in fowl. He studied chicks that had ‘spirochaetosis’ and at a certain stage the spirochaetes had disappeared from the blood, but he looked at their liver fluids and he saw them swarming there in the liver in high numbers, shimmering and moving vigorously. I wanted to look at that one day, could that be possible?

In 1912: Hindle described a cycle of spirochaetes going in cells in tick, not the human, in blood in the tick and degrading into granules and the granules leave the cell and grow out to become spirochaetes again. Normally, you would talk about division of spirochaetes and division is like ‘cutting a worm in two’ and then they grow again and that is not very effective (as a means of reproduction). But if you cut a spirochaete like many slices of a sausage then you have an enormously effective reproduction. He saw these things in1912. I have an old fashioned microscope and I thought to myself, maybe I could have a look at some blood too, I had no staining material etc., I didn’t have anything….

In 1914: Nicolle and Blanc did another study on Relapsing fever (Borrelia recurrentis) Borrelia patients. Here they took out the lice (that had) eaten from these patients and they looked at the blood in the gut of the lice. They saw that no spirochaetes were present, for Relapsing fever Borrelia. One knows that during the crisis (the relapse phase of Relapsing Fever) there are spirochaetes visible in the blood. Between relapses, in the afebritic phase, the is no fever, the patient feels pretty well between relapses. (At this time) there are no spirochaetes present, and they took the blood from the lice at a time with no spirochaetes and injected this into animals and the animals produced a spirochaetal disease. So they concluded that the agent must be in an invisible form in the blood between the spirochaetal phases. Still talking about Nicolle and Blanc, another observation they did, when they looked at the blood, when they took the blood after a meal. In the first One to Four days there are no spirochaetes visible in the blood and it is not ‘violent’, in 4 – 6 days a few spirochaetes are seen and they are a bit ‘violent’, 7 – 9 days later there are more fine spirochaetes and they are ‘violent’. After 10 days there are many spirochaetes but they are grown-up (mature) and ‘not violent’. This is a very important observation.

In 1914: Sergent and Foley did something similar. They also found that the agent of Recurring Fever (Relapsing Fever) were in an infectious form in the blood for 6 – 16 days, but the mean was 8 - 10 days, concurrent with the afebritic phase. Fantham did some drawings of spirochaetes in 1907, where he drew from a camera lucida (Marie explains that this is an old-fashioned drawing technique in which projection of light gives the ability to draw a specimen directly from life). At that time they had a lot of discussion about how to categorise these spirochaetes and how many turns they had, how long they were, how thick they were, how many coils they had, but Fantham said ‘that is meaningless’. He said this because they were living structures and they can move like a snake in a zoo, they can coil and uncoil and go fatter or go longer or less long and this drawing shows that. He also shows that they get granular and then they break into these granules (see drawing).

1918: Leichmann – His work too, but I won’t go into that

In 1948: Hampp had already done Electronic microscopic studies of the cyst form from different spirochaetes including Treponema pallidum, the Syphilis spirochaete and Borrelia, not Borrelia burgdorferi, it wasn’t named then. He wrote that the electron-micrographs demonstrate that this explanation is wrong, that free granules are definitely a phase in the development of spirochaetes, although it is not possible to discover from these micrographs that the granules are germinating units, but their constant rhythmic occurrence in living culture suggests this possibility. Further support for this hypothesis is provided by the fact that cultures up to one month old showing only refractive granules by dark field observation have invariably driven normal growth when transferred to fresh medium.

Persistence is the keyword. The old guys had to look down the microscope for many, many days without leaving their observations, because they had to stay looking in order to observe the changes because they are not quick. The other guys, who were less persistent and they only looked for a couple of hours, they didn’t see what happened and they decided “it doesn’t exist”. This isn’t true. Other workers did see the granulation process but they said ‘well, they are just going dead’ because they degrade and this was also (the case) when you are looking at fixed stains, you can’t see any changes. The result was that this theory of a complex life-cycle for spirochaetes was considered wrong and discarded and none of us here today have been taught about these findings.

Today I will show you that such structures are both alive and moving. Yes, but before we get to this, there are some new observations that support the old observations I think. You know that Borrelia can go intracellular and the intracellular location protects them against the effect of some antibiotics. I have made a little list of these theories here, you can all see it on the internet (where I have posted it). We also know the alternate cystic form, spiroplasts, L-forms or whatever they were named, formed when the Borrelia encounters a hostile environment. In the presence of antibiotics, in the presence of antibodies, there are more of these works and when antibodies form. This was the first work to show that the cyst forms are already in the skin. In 1999, McDonald found cysts and spirochaetes together in the brain of patients dying with Alzheimer’s disease. (When they have) lack of nutrients – Ellman starved the Borrelia and they went into the cystic form. And not the least, Brorson’s work when he put them into spinal fluid or distilled water they quickly within 24 hours found the cystic form and when they were put into richer medium they reconverted back to the spirochaetal form. There are pictures of the alternative forms of Borrelia on my LymeRick website. Last and most important work was in proving Nicolle and Blanc’s work from 1914. They took cysts of Borrelia garinii and injected them into mice with no spirochaetes present and they could later find spirochaetes in the hearts of these mice. (Gruntar). So they proved that the cyst form is infective.

Timing is important when considering this illness. Patient gets cyclical symptoms and as I have already mentioned there is timing in Relapsing Fever Borreliosis. It seems that when patients are symptomatic they are reacting to the spirochaetes and the body does not react so much to the cyst form. Borrelia burgdorferi, is also a cyclical, relapsing disease. This is explained by differences in cyst growth velocity found between young, at about 9 days, and the old that had reduced metabolism to very low for a longer period. (Brorson). This work also writes that the cyst resembles the L-forms described by other researchers. What is most important is that the biological activity of the cystic form was confirmed by the step by step development of the normal mobile spirochaetes in gel or Kelly medium. And also implicated by the presence of rna in 5 week old cysts due to orange staining with acridine orange (a fluorescing dye) which shows () and shows the timing. I had already thought about the timing of the illness.

I had noticed in myself and other patients – a sense of a monthly cycle, I also sensed a short cycle. In fact a patient had started to make notes on this, without my telling him to – he followed 7 or 8 symptoms, and he gave them a severity score, and we followed the course of his illness – a three week cycle. I have developed an Excel programme in Danish – long, long lists of symptoms. I asked this patient, I asked him to score them 1 – 4, and had to give grades in between. ( I described to patients how to do it, so that the score was a graduum). All symptoms scored and we sum it up on a table. And you can see there is a cycle. This is a patient, 17 was his total symptom score – he has a cycle, and here he forgets to keep it. Then he had increasing symptoms again and wrote it again and during a warm August and he felt a little better – then treatment - Herx’ing and then began improving, then there is a break in his graph and it jumps and he gets near a monthly cycle.

An infectious antigen which relies on transmission by a blood-sucking vector but which can only be found visible in blood at intervals of about 9 days would not be effective if it was not there, but it is explained by the equally virulent and infective ‘invisible stage’. But actually what others thought was invisible is not invisible!

We have to look for granular structures. Or other cyst forms, the structure will react positively with Borrelia antibody tests from Bowen (laboratories).

Lets look at a video to look at the spirochaete – from 1997 – (Marie shows video clip).

You see bright, small, dots moving around outside and you see this spirochaete getting ingested by a white cell so called tube-phagocytosis. It’s a long video, I wanted to show you that Bowen put one with the dark-light appearance you can actually see it here
I also made a video, before I show that - I need a couple of patients because I am going to have a demonstrate live also.

Another patient, she was number 11. She has Borrelia burgdorferi, Babesia, Erlichia and this was confirmed by Walter Tarello and myself. At first we gave her Doxycycline 4/5 monts and this had no effect at all. Ill since 1973. Then I read an article that said Metronidazole has an effect on Babesia with Azithromycin, two drugs together, both are able to go intracellular and across membranes, both would be good for Borrelia also, a combination that could work for her? What doses? –it might be a combination of doses working or doses normally used and recommended (except for the duration of treatment). Then in about here in April she went to this treatment and within 2 to 3 weeks she felt much, much better and she enters the monthly cycle and got later still one week between. Here you can see - nicely presented the flares. You can see – very few symptoms. Her score here is ten points total and here she suddenly has a month without any flare, she had one here a little one, she after she went onto the monthly cycles she has only taken therapy at the first symptoms of a flare. ( I’ll come back to the reason for that).
All antibiotics interfere with metabolic processes – protein synthesis, dna synthesis and formation of cell wall etc., and in inactive Borrelia bacteria antibiotic cannot do that, they are sleeping and the drug cannot hit them. So you give the drug and it is toxic to you perhaps and it has no effect on the Borrelia.

Usually have to have high doses for drug to go into the privileged sites like cns, tendons and inner eye, in sufficient concentration to suppress the growth of bacteria. We know from old work and relapsing fever patients, Borrelia can persist in such places and stay dormant for many years. You can refer to my website. Cases of verified relapses of Borrelia, microscopical, culture, PCR verified cases and about half of these patients are so negative, never had the energy to count them, too many simply. And I thought -
Why punish yourself with antibiotics when they don’t work during the pulses?

I have found when I treat with continuous treatment in the first three months until the patient goes to a monthly cycle then they can treat themselves with drug treatments when they feel the need to. Then (if they do this) they have the succes – and improvement.
What I saw before was people took treatment for 2 – 3 months – improved and everybody then told them ‘it was dangerous to take antibiotics for such a long time and they must stop’. Also it is expensive, they reached a lag phase and they only had this constant process and a flare now and then and they didn’t tell me and they stopped treatment at this point and they always relapsed. They will tell and I will tell of all these relapses
- flares. And it seems that with the pulse therapy, you can remain stable on the level that you have achieved and its cheaper and less toxic to say the least.

I need somebody who is not ill and I want to make a comparison and one who has never had anything, or in the last 10 years Are there anybody in here who can say they are totally healthy and never had a tick-bite (laughter). Someone not married to a patient. (Someone volunteers). Ok we’ll take you.

I saw a few patients yesterday and I made a little test how good they were and I picked Marie-Francoise because I saw something in her blood. But probably if we don’t succeed I can pick any of you I think.
Marie takes blood from volunteers’ ear lobes. While I am doing this I’ll show you little films. Marie shows a slide - saying granules and granulated cellular structures. My camera is not the best. But you can do it with anything even my little one. It is really back to basics. Then some round projections through her camera apparently showing something granular
- A free clumps of granules, no surrounding membrane, slight movements in clump.
- Another slide you can look at - granules moving like pearls on a string, what is it?

Q - What magnification is this? Marie says it is x1000.

Can see something crawling here, this structure it is rare to see them move fast, usually have to sit and look, at least half an hour or so, be still for a long time, its really hard to get something and then suddenly something happens. I don’t know if I can show you today. I ask - is it within the cell structure or isn’t it, can’t see the neutrophil
I (Marie says) think it is in the cell. I had to for myself.
In 1911, old guys described that they enter into the red blood cells also.
I think they can use any cell. This assumtion is supported by the fact that such granules is stained with the Bowen RIBb test.
Marie shows a Phase contrast image of the same cell showing Borrelia burgdorferi
Now I try to switch over (switch between light microscopy and dark field microscopy). Then Marie projects a patients view of rbc (not live) and she shows something in rbc stain? First of my smears, I diagnosed Babesia before Bowen, I knew I could find it in a ring form. Now you have seen it.

Let’s look at Marie-Francoise’ blood (and this is projected), live and the rbc are floating around - (lots of views),nothing, nothing, nothing, left a little too long and they (rbc) were conjugated. Yesterday there were more – this is the problem, you can’t just see things – eg you can have Babesia and have them bursting in front of you and just after it is impossible to find anything. Then a few days later find them again.
Then just before an expected flare you can find something

I had six or seven people yesterday evening that were full of them. Marie Francois tell your story and present symptom level and everything while I am looking…. (The very interesting patient stories have been cut in the interests of brevity).

I have seen 33 patients so far and I have made an overview. Well, some have many tick bites. I was the first patient, I had Walter Tarello to make a blinded control and on 12 of the patients. All results are listed below each other
Babesia in red, Human granulocytic erlichiosis or other suspicious inclusion in the white blood cell is shown in bold.
All positive for Bowen test and stain for Borrelia and positive for granules in the blood.
and we can find some in this slide later and have a look at the pictures. Bowen does phase contrast (microscopy) andimmuno-fluorescence stain. Marie shows the Bowen slides - more sophisticated equipment means they can switch forth and back and hoto same thing with the two different methods (light and dark ground microscopy).

This is some of the pictures of my blood from Tarello. He also found some micrcocci, small red dots, the thing about micrococci is that they change metachromatically, red and blue, up and down through the magnification so you can easily spot them.

All pictures from Bowen that I have cut out, immuno-fluoresence stain and you can see how different they look at different stages. (Q asked from audieence ‘Is Bowen going to be fully licensed?’ Marie- ‘ Well they have sent in something for publication, then a review but a year or two before it is accepted, it is not good. These are some of my own pictures. I don’t have the most sophisticated equipment.
You should take care, this was Erlichia, but I don’t agree, Erlichia is supposed to be in a so-called morula.

To sum up:

All these patients have typical symptoms of chronic Borreliosis.
Symptoms, you have heard about weekly or monthly patterns as Dr. Burrascano describes – cyclic activity.
No other signs when is the tick bite – no Ecm. No positive serology. But patients were enrolled by having typical symptoms from their diary. All had a positive result from the Bowen laboratory. I have made a one page demonstration of some of the structures seen.
These are actually my own two, one nearly ready to burst, one the way and one that has burst and only the exterior is left. All patients have similar granular structures, seen and filmed Age span 5 – 55 years old, mean age 35 years. One third of the patients acquired the infection before the age of 40. 95% were sick for more than a year of disease duration, but 5 months to 33 years is the time spans I have recorded so far.
Only three patients were sick for less than a year.
In the patient who was sick for 34 years – it was very interesting, a neuropsychologist had written about this patient, write since 1983. They wrote “11 times a year she has recurring flu-like symptoms, she got blood spots on the skin, she was flu-like and she felt a headache, acid feeling and joint pain – a very poorly patient and ill and it lasted about a week.” I have never seen so good a description of chronic Lyme Disease! From a neuropsychologist written 3 or 4 months before Claus Hansen published his thesis.
3 or 6 per cent were negative for Borrelia antibodies despite typical history(borreliosis) and 27% Em patients had no antibodies. (!)

Ten our of thirty three had Babesia. Looking at Babesia - intracellular ring forms are never a normal finding. It’s not easy to discriminate between thrombocytes and the parasites. But thrombocytes, they don’t have a ring form, thrombocytes a bit more red and others a bit more purple staining. In high magnification you can see whether the form is intracellular. You can alter the focus and see over, under or in the middle of a cell. So I have always recommended to do the microscopy x1000 at least.
I also have a Plasmodium vivax slide and it looks much the same. Babesia resembles Malaria. Patients with this may have more and worse symptoms in this Babesia and Borrelia co-infection:
If I give doxycycline or amoxycillin alone it takes a long time to get half way to getting to a cycle, compared to those with only Borrelia.
But when I give the metronidazole/Azithromycin they follow usual pattern of six weeks to the basic level of having the monthly cycle, so it seems that of having the metronidazole does something.

Q – Is this information available generally?
MK – no, but on my Lymerick web-site. I have referenced the sources.
And I have added something re: the erythrocytes from another source, what is the normal distribution and what you name red cell abnormalities and more about Golgi bodies. A little about the small, dark spherical granules inside the red blood cells, thought to be remnants of a nucleus (golgi?) but I am not sure it is, they are larger than normal red blood cells and have blue cytoplasm.

This is stain for reticulocytes. In Bowen test – polychromacea.
Differences in colour and these cells are a little bigger, but can see these erythroplasts in the normal blood, but golgi bodies are usually located into mature cells, so I think there is something there… This was probably inside normal blood and haematologists described normal blood thought this was a normal finding and I thought it could be Erlichia because it is not a normal finding.
Slide 9 – 33 Had abnormal inclusion in white blood cells.
On the net there are a lot of pictures of links of parasites and bacteria and because when you look you must know what they look like. White blood cells are phagocytic and ingest all sorts of microbes so when you see a blue dot you cannot distinguish them. Marie then describes another series of slides. Audience asks is this on the website? There is a CDC webpage where you can use the pictures.

Q – Website?, it is on the first page of my slides. Not up yet. (http://Lymerick.net)
It is on mmi and eurolyme when it is up.
Spirochaete can survive intracellularly – also in immune cells as has already been mentioned today.
If they manage to wipe out all the white blood cells that are going to fight them then they are really going great.

Chronic infection, all the symptoms and diagnosis in the patients,
Often misdiagnosed as FM, ME, CFS
Six per cent were described as FM, ?cfs despite the facts of tick-borne infection history going to chronic tick-borne infection causes their disease
Three patients were diagnosed with cfs without the relevant testing!
Many patients were told, without proper evaluations, ‘it is just stress, psycho-somatic….. many patients in the medical system were refused testing for Borrelia and co-infections, even though it was possible to do these tests in Denmark at the time. It is especially relevant if patients had positive history and serology had had a response to treatment and then (stopped) and have a relapse!
Many had Em diagnosed as Borrelia but doctors overruleld with when serology is negative (the diagnosis denied), despite there are plenty of positive cases. I want to point out an example of a lady got carditis and was treated for six weeks with doxycycline and it disappeared. A couple of months later, a relapse, cardiac abnormalities, ECG, joint problems, 2 weeks Ceftriaxone and she improved a lot. Then later she got again symptoms – trigger finger due to inflammation and she had finger operation - pull out the tissue and the spirochaetes were in the tendon. And when they tested her, after the first treatment she was sero-negative, tested her specifically for the cultured strain she was still negative but had a t-cell response to the strain – and I use this word - they ‘cannot explain’ this patient. Well just a few months ago, many doctors said undocumented things to patients could easily look and see was false, because today the patients find the information on the internet. Whereas doctors often avoid the internet – say it is rubbish etc. but where there are many good things.
Why can’t all patients be examined as thoroughly as Howells patients and patients won’t accept the false diagnosis? Are doctors too stressed to care and why won’t they acknowledge the existence of chronicity in Tick Borne Illness?

Chronic tick-borne illness:

It is doctors more than the patients, because we learn to live with it
patients with chronic illness, whether tick-borne or not, deserve the best services, because it is always worthwhile to test, diagnose and treat even if it is not always easy. And all doctors should have a simple microscope, - no big deal and when they find such structures go to send on to an appropriate laboratory etc.

My Pilot project of 33 patients and not all are followed because they don’t want to keep a diary.

I only test patient patients (followed 10) because I need them to keep a diary, see if structures in blood correlates to symptom level and I can only do that if I have the score. When patients come to me and I can see untreated and highly symptomatic, I find a granule structure in every cell. During a few months treatment, then I find one or two only in the whole slide. I also find one or two in patients between the flares, we still don’t know how it is correlated but everybody can look for it who has a microscope.