Video-microscopy and
pictures of Borrelia burgdorferi and other spirochete like structures links
collection
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Survival
in Adverse Conditions (2001)
An organized collection of photographs and
quotations selected from studies finding evidence of alternate forms of various
spirochetes, dating from the early 1900's through the present.
Written permission to
publish this article in LymeRICK from the author, who
wants to remain anonymous.
Alan
MacDonald – USA
Culture of Borrelia burgdorferi (B31)
-
stained with fluorescence
marked
specific Bb-antibody; done circa 1985
“Of
Course you have my permission to include any of my images on your web site. I
Do not desire to copyright the images, because so many in the Lyme Borreliosis
community have viewed these images over the 20 years since they were created,
that it constitutes
a de facto acknowledgment and I know that my photomicrographs exist to serve as
tools for education, and not for commercial purpose.
The image of the "cystic" form was obtained from very aged cultures
of B31 from the ATCC. After the motility of the borreliae
ceased, the "formes atypiqes"
appear, and there forms are very diverse. Cystic forms which contain granules
inside of the cysts are just one of a
myriad of atypical forms that Bb
may assume.”
[wrote Alan MacDonald to Marie Kroun in personal e-mail about
this picture on 20050830]
Matti Viljanen, M.D., Ph.D.,
Professor – Finland
Tube phagocytosis of Borrelia burgdorferi
(fast Internet
connection Real Player Movie 3 Mb,
Above links to – and here is the text from –Viljanens own website:
”The
video is a digitized version of the original video, from which the still images
in figure 2 of the article "Tube Phagocytosis,
a Novel Way for Neutrophils To Phagocytize
Borrelia burgdorferi" by Juha Suhonen, Kaija T. Hartiala and Matti K. Viljanen (Infection and Immunity, July 1998, Vol. 66,
No. 7) were captured.
The
video is presented with the kind permission of American Society for
Microbiology. The video presents a novel type of phagocytosis
described in the article mentioned above. The original video was produced by
observing and recording the interactions between human neutrophils
and Borrelia burgdorferi, the Lyme disease spirochete, using dark-field microscopy
with video technology. The digital version was made by Antero Lehtonen.”
Marie Kroun, MD – Denmark
18 MB, 2 minutes video first presented at the
conference in York
“Pearls on a string”
spirochete like structure moving inside a celular
structure from microscopy of blood from pilot project patient#18.
More of MKs most
recent videos of moving “granulated cellular structures” (a pure descriptive
name ~ “cysts”), taken with a USB-camera:
Bresser PC-Microocular-II (cost only 99.95 Euro):
The videocam fits into
the ocular of most microscopes.
0018-20051213 (WMV, 23 Mb) — note this is the same
patient as above!
0049-20060704 (WMV, 50 Mb) — video from
project patient # 0049 (NEW)
Should you happen to see
something like this in an unstained wet blood drop microscopy, then you should consider
doing supplementary diagnostics, like microscopy with specific immune stain for
B. burgdorferi.
So far
(Jan 06) 45/45 patients with similar structures found in their blood and
displaying current symptoms compatible with having active Borreliosis had
positive outcome of a specific immune stain for Borrelia burgdorferi, done by
Bowen RTI, see below!
Unstained
wet-drop microscopy can be done on a fresh ear prick blood sample, on
anti-coagulated full blood (EDTA, citrate) or maybe even best on a sample from the Buffy-Coat
fraction.
See how to make buffy-coat smear video.
Some references on
buffy-coat microscopy for parasites: QBC Malaria, Paralens, and Google and PubMed search.
Microscopy of buffy-coat preparation (wet drop or dried smear) could of course
also be combined with use of a highly specific immune stain for any suspected
pathogen, for more specific diagnostics.
Once dried the (buffy-coat) blood smears can be
stored for several years!
A German colleague and good friend, who is a tropical diseases specialist, gave
me a 20-year old blood smear from a malaria patient, we stained it and looked
in the microscope and really couldn’t tell that it wasn’t a recent smear and
malaria parasites were easily visible in it, many more than is usually the case
with the tick-borne cousin babesia!
ALWAYS TAKE PLENTY (buffy-coat) BLOOD SMEARS
whenever you have the chance, since
the cost of the glass slides are low, they don’t take up much space if you
stack them in the same box you bought the slides in, they can later be used for
trying other stains for comparison, for using additional (more specific immune)
stain or you might even scrape blood off the slide for doing PCR, in order to
get a more specific diagnosis than just “ringforms seen” or “morulae-like
inclusions seen”, use it for education of laboratory workers and microscopists
etc.
– you can really never get too many good smears with parasites in it!
Bowen RTIs procedure of doing direct fluorescent antibody test for Borrelia
burgdorferi was patented in Jan. 2005; see the US-patent description for all details about this test.
The main advantages of Bowen RTIs Q-RIBb test are:
1. Documentation by picture(s) of any microscopic finding, so you can
compare your own microscopy findings to that
2. Quantifying - by titration - the number of
structures reacting with added specific antibody against Borrelia burgdorferi.
As stated in the patent description Bowen RTI use a commercially
available anti-Borrelia burgdorferi antibody from
Affinity purified polyclonal antibody to
Borrelia burgdorferi made in Goat and labeled with
fluorescein isothiocyanate (FITC).
Isolated from a serum pool of goats immunized with heat
killed whole cells of Borrelia burgdorferi.
The
antibody is highly specific for Borrelia burgdorferi.
Cross reactivity to Borrelia hermsii, Borrelia coriaceae, and
Borrelia anserine has been minimized through extensive affinity adsorption.
Product is in lyophilized form. Each lot is tested to assure specificity and
lot-to-lot consistency using KPL's in-house ELISA
assay.
Andy Wright, MD – UK
AndyWright2004-640.wmv
(52 Mb, 4 minutes) video of spirochetes
in more lenghts, granules,
a moving granulated cellular structure, thus illustrating all the phases
complex spirochetal lifecycle drawn on page
So far (June 2005) 98/98 of Andy’s ME/CFS patients with such structures
in their blood tested positive on direct fluorescent antibody test for Borrelia
burgdorferi !
Bela Bozsik , MD – Hungary
Dualdur® movie
70 Mb, 11 minutes dark-field microscopy
video of Borrelia burgdorferi spirochetes in blood, showing the shedding
of ‘granules’, budding of the spirochete at the end and centrally, the
formation of blebs and transversal division of the spirochete.
Dr. Bozsik adds a
reagent to the blood named Dualdur, on which he holds
the patent, which immobilizes blood cells, but do not disturb the spirochetal movements.
Dr. Bozsik kindly gave me his permission to present both his lectures DIAGNOSIS and THERAPY held at the Sheffield 2005, UK LDA conference on tick-borne infections
Both
presentation are made in PowerPoint 2003; some embedded videos from “The
Passion of Christ” and “biting tick” shown at the conference does not seem to work
properly in the this PPT?
The latter --
tick bite video -- show us how Ixodes tick enters its hypostoma
directly into a small skin capillary.
The implication is that in case the tick is
systemically Infected with Borrelia burgdorferi the spirochetes will be
injected immediately and directly into the blood-stream with the tick saliva,
and for those ticks not systemically infected from start, spirochetes migrates
within 1-2 days from the ticks mid gut to the saliva glands – in any case Borrelia
is injected directly into the blood stream during the blood-sucking! – thus
Borreliosis is a blood infection right from start, explaining how spirochetes can spread
early – before occurrence of EM - into all tissues and compartments, like the
central nervous system within only two weeks of the tick bite. Therefore
treatment of early borreliosis should aim to reach a
sufficient concentration of antibiotic – i.e. over the minimal inhibitory /
bactericidal (MIC / MBC) concentration - within the CNS right from start, in
order to prevent survival of Borrelia in CNS and reduce risk of later recurrent
neuroborreliosis, due to initial under treatment!
Dr. Bozsik
found – data extracted from his Diagnosis
lecture – that:
- Spirochetes could be demonstrated in BLOOD by dark-field microscopy during all ACTIVE
stages of pathogenesis!
- 107 of 143 (75%) of the cases with live (moving) spirochetes
found in their blood by dark-field microscopy, were confirmed by real-time PCR to belong to Borrelia
burgdorferi sensu lato:
66 (61.7%) B. burgdorferi sensu strictu
20 (18.7%) B. garinii
6 ( 5.6%) B. afzelii and
15 (14.0%) other Borreliae
Many of these cases were also confirmed by monoclonal antibody stain for Borrelia burgdorferi with anti-ospA and anti-flagellin kindly donated by prof. Barbour - One third (1/3) of these patients with
LATE Lyme borreliosis were SERONEGATIVE(*)!
*) A considerably high number of patients with
proven late chronic Borreliosis, i.e. where persistent Borrelia
infection was proven
by ANTIGEN DETECTION, either by CULTURE, PCR and/or MICROSCOPY (sometimes aided
by specific immune stain) for Borrelia burgdorferi has been shown to test
NEGATIVE on a variety of BORRELIA SEROLOGY tests!
Oksi et al. J Clin Microbiology 1995 Sep; 33(9): 2260-4 (PDF) published the results
of 3 different SEROLOGY tests done on CULTURE and PCR proven LATE (> 3
months after acquiring Borrelia infection) BORRELIOSIS cases.
All 41 patients had symptoms
of Borreliosis for at least 3 months; 78.0% > 6 mo, 53.7% > 1 year!
Number of positive Borrelia burgdorferi
ANTIGEN results:
12 were positive on CULTURE, 39 were positive on PCR – 10 were positive on BOTH
CULTURE & PCR!
Performance of 3 different SEROLOGY
tests used on all 41 patients:
1.
2.
3.
Overall results of SEROLOGY
tests on all 41 patients:
19 (46%) were only borderline
or weakly positive
7 (17%) were SERONEGATIVE
3 serologiske test; 19 havde kun
svagt pos. eller grænseværdi antistof niveau; 7 (17%)
var SERONEGATIVE på ALLE 3 tests! - Resultatet af FL-ELISA (DAKO, Glostrup,
Danmark): 6 dyrknings pos. + 18 PCR pos. = 24 af 41 (58%)
med SEN symptomatisk Lyme borreliose var
SERONEGATIVE! - dette ELENDIGE resultat af bekræftes i andre undersøgelser
f.eks.
Marie Kroun’s
pilot-project presented at medical conferences in
36% of 33 patients tested SERONEGATIVE on the danish serology test for Borreliosis (DAKO)!
NEGATIVE SEROLOGY CAN NOT BE USED TO EXCLUDE PERSISTENT ACTIVE BORRELIOSIS!
Unfortunately many doctors
does apparently not know this FACT, because they still use a negative serology
test for Borrelia burgdorferi to falsely out rule Borreliosis, leading to false
diagnosis, no treatment, no cure for a chronic disabling and sometimes life threatening
spirochetal infection, that can be treated successfully
with antibiotics in most cases, when ANTIGEN diagnostic measures are being used
and correct diagnoses made EARLY in the course in infection; why
saving these patients and returning from !
If
YOU have videos or pictures in the same category that you want to show the
world, please contact me!
Marie Kroun, MD
kroun (at) ulmarweb.dk