Borrelia Spirochete Morphology in Peripheral Blood part 6

Discussion

It is not claimed that all of the elongated agents in the images and videos are spirochetal, yet many have characteristics that when correlated with the known morphology of spirochetes and the donor's health and diagnosis; as well as the successful antibody staining of 2 donors, supports the idea that they are in fact spirochetes. The results were variable but it was shown that these forms could be cultured and observed in samples prepared by the donor. The quality of the microscopy imaging was dependent on various factors. The quantity of blood cells, ratio to and mixing with the BSK and depth of the sample all affected the microscopy. Nevertheless, donors who had little or no previous practice managed to prepare slides that produced interesting results.

Problems encountered with Kit 1 were mainly around the size of the blood-drop. An ideal amount would spread to cover 50% to 70% of the area of the coverslip. This was difficult for donors to judge, requiring accuracy to a few microlitres. Too much blood would seal the edges of the coverslip and left no room for the addition of BSK. Too little blood resulted in desiccation of the sample and poor mixing with the BSK.

With Kit 2 BSK was added by the donor making the process more complicated. This slowed down the preparation and meant that blood and BSK were exposed to the atmosphere for longer and increased signs of contamination were noted. Mixing of the blood with the BSK was sometimes incomplete - donors were asked to stir the mixture with the pipette tip for 2 to 3 seconds which proved to be too short, sometimes leaving obvious clumps of blood which resulted in a deep sample. Some of the slides containing rather deep samples for microscopy might have improved with further blotting of the slide prior to sealing the coverslip. The Donor Instruction Sheet requires improvement in these respects.

Donors were people diagnosed with M.E. and/or LB. In these illnesses a phenomenon described as 'sticky blood' can occur. Dr Byron Hyde remarked: "It is well worthwhile for all physicians reading this definition who have an interest in M.E. to examine the Internet for Hughes Syndrome…. a vascular syndrome also called Sticky Blood Syndrome, closely parallels the definition of M.E." (18)

Dr Charles L. Crist is an MD with experience of treating Lyme borreliosis, he observes on his website (http://drcharlescrist.net/Borreliosis/Hypercoagulation/ Accessed 26 November 2013.): "Hypercoagulation, or thrombophilia, may be defined as reduced capillary blood flow or a greater tendency than normal for blood to coagulate, or clot. Of approximately 900 borreliosis patients that I have tested, 90 percent have hypercoagulation. Comparatively, only five percent of the general healthy population has hypercoagulation."

'Sticky blood' does not spread well beneath a coverslip and can form a thick film of which the outer edge dries-out quickly. It does not mix readily with the BSK and can result in a sample that is inconveniently deep for microscopy.

In addition to the problems mentioned, persons with M.E. or LB can have significant problems with coordination and fine motor control, impaired eyesight and difficulties with concentration. Some donors had to make special arrangements to post their samples. Given the severity of the illness of some donors it is remarkable how well they managed. It was found that the donors were interested and enthusiastic about trying the experiment and that with little or no previous experience they were often able to prepare a slide suitable for microscopy.

5 of the donors in this experiment had previously provided 2mls of blood in an EDTA collection tube. This method permits control over the preparation of sample slides and enough sample to start long-term cultures and conduct other experiments; making it a far better method than the kits tested here. Nevertheless, this simple experiment gave some who are virtually housebound by their illness an opportunity to participate in an experiment that has produced results.

Variations in the results occurred even from individual donors which suggests that the method is not optimum; it might also indicate that detection of low numbers of spirochetes would be even more variable. The shortest duration of illness amongst the donors was 5 years, with 8 of the 11 ill for more than 19 years. It is unknown whether this experiment could culture spirochetes in people who have been ill for months or even a few years.

The results shown in some videos appear rather shocking. It must be remembered that this is not the natural state of the donor's blood but shows an artificially created environment aimed at encouraging spirochete growth. No attempt was made at counting cells and minimal donor details were collected. Yet an impression was that the severity and length of the donor's illness did not correlate strongly with spirochete numbers in this experiment which looked only at peripheral blood cultured for a limited period. Rather, the ease with which spirochetes were found and their numbers seemed to correlate more with the characteristics of the sample; i.e., a shallow sample with a reasonable density of well distributed cells seemed to be the factors that reliably provided an opportunity to observe spirochetes.

Blood put on a microscope slide undergoes numerous stresses which can damage the cells and create artifacts. Yet 24 hours or more after a slide was prepared, samples often appeared perfectly 'clean'. Cells had a normal appearance and no spirochetes and no collagen fibers or anything else that might be visually interpreted as being a spirochete were observed. The spirochetes have come from somewhere and logic suggests that they either grew from spheroplasts present in the plasma and/or attached to the surface of blood cells; and/or that they were inside cells and emerged from them. It has been suggested that agents like those observed are the product of cell membranes disintegrating and this theory would accord with the progressive increase of the spirochete-like agents over time. However, it does not accord with the fact that within a few days of their appearance, the spirochete numbers decline rapidly having converted into numerous coccoid forms. If these long-agents were artifacts of cell decomposition, one would expect their numbers to increase until the majority of cells had broken down.

Limitations of the experiment

Cell counting was not attempted. No healthy controls were included nor people with recent infection. The spirochete species is unknown.

Conclusion

Donors with long-term illness and diagnosed with Lyme borreliosis and/or M.E. can prepare mini-culture slides from a fingertip blood-drop using a simple postal kit. This allows direct observation of spirochetes reasonably consistently with darkfield microscopy within 5 days. When antibiotics were being taken by the donor at the time of sample preparation it did not prevent the observation of spirochetal forms. Long-term culture and fluorescent antibody staining can further support the conclusion that spirochetes are present. The replication of some of these experiments by qualified persons under laboratory conditions and with healthy control subjects would be of interest to the medical and scientific communities and the many patients that could be affected.

Acknowledgements

Sincere thanks are due to the donors whose samples and the trouble they took to prepare them made this experiment possible.

Navigate this article:
Borrelia Spirochete Morphology Introduction
Borrelia Spirochete Morphology Results
Borrelia Spirochete Morphology Extended Results
Borrelia Spirochete Morphology Supporting Evidence
Borrelia Spirochete Morphology Fluorescent Antibody Experiment
Borrelia Spirochete Morphology Discussion, Limitations, Conclusion and Acknowledgments
Borrelia Spirochete Morphology References

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