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» LymeNet Flash » Questions and Discussion » Medical Questions » The Microscopy Thread (Page 18)

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Author Topic: The Microscopy Thread
TNT
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quote:
Originally posted by Lymedin2010:
Anything is possible, but you can also see the bart inside the rbc with the hemoglobin still intact & not compromised. So both the hollow one & the full rbc have what to me look like bart.


Do you see any stained spirochetes in your blood? I only find a few of them with staining & they are questionable in appearance, as in they may not be spirochetes either. So I wonder about that too since I have seen stained spiros in slides online. I was once led to believe that any SOP in the blood meant that it was foreign, but after my acid & basic (baking soda) experiments I no longer hold to that belief. Shedding of the rbc wall can also produce SOP's.

I look forward to your tick ventures, can't wait...how odd what us Lymies get excited about.

I do not see spirochetes in my stains like we see in our live blood. Relapsing Fever spirochetes stain well, and those are probably the pictures you see online of stained spirochetes. For some reason, Bb spirochetes do not stain well with Giemsa. I don't think Syphilis spirochetes stain with Giemsa either. I have only seen a few objects in my stains that could have been spirochetes, and they were debatable.

Unfortunately, I doubt I will have the time or energy to pursue staining insect contents any time soon. But, it's a possible interest for the future.

Disintegrating red blood cells can definitely form string of pearls, but those pearls are bigger than the pearls of spirochete SOPs! Isn't that what you have found?

[ 03-29-2017, 03:17 PM: Message edited by: TNT ]

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Lymedin2010
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Yes, I found out larger SOP's can form from rbc cell walls & I wonder about the smaller ones too, as I don't think there is a limitation to how phospholipids can form below 1 micron & lower...just think of micelles fat droplets & how small some of them can get, which are basically spherical phospholipids.

I know spiros can & will be in our blood, and we have seen some awesome cyst forming videos. I think sometimes we might confuse one for the other though & hard to tell at times. I use aggressive movement of the spiro now as a better judgement to distinguish from false spirochetes. A darn electron microscope can easily solve the questions if we had one at hand & I was considering looking into buying some EM time.

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Lymedin2010
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Damn, check this out.
https://www.youtube.com/watch?v=riVHNOPTjyo&t=14s

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Lymedin2010
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Here is the rest of the Bart references from the previous time, for anyone else interested.

MORE ON BART VACUOLES IN THE BLOOD, forget about species this is to learn more on if bart in general can be in the blood & within the rbc's & whether it can form vacuoles inside the rbc's...continuation from last time:

1) "Deep invaginations containing bacteria are commonly seen, and membrane fusion at the necks of the invaginations leads to the formation of intracellular vacuoles containing bacteria. Fluorescent compounds present externally render the vacuoles fluorescent and, occasionally, lightly fluorescent cells are seen, suggesting that the vacuoles sometimes rupture to admit the bacteria to the cytoplasm."

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC260167/

Full pdf:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC260167/pdf/iai00098-0083.pdf


"FIG. 3. Deformation of erythrocytes as seen by scanning electron microscopy. (a) Clumps of bacteria and (b) single bacterium bound at indentations. (c to e) Deep pits which result from bacteria pushing into the erythrocyte membrane. (f) Membrane of the erythrocyte apparently pulled up from the cytoskeleton and twisted."

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"FIG. 4. Transmission electron microscopy of thin sections of complexes of bacteria and erythrocytes. (a) Bacteria lying clearly outside of the erythrocyte. (b) Bacteria apparently within an endocytotic vesicle"

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"FIG. 5.Micrographs of erythrocytes preloaded with calcein and infected with bacteria. (a and c)Visualized by dark field and (b and d) visualized by fluorescence."

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______________________________________
2) "We identified a Bartonella quintana strain by polymerase chain reaction amplification, cloning, and sequencing of DNA extracted from lysed erythrocytes and cultured colonies grown from peripheral blood collected from a captive-bred cynomolgus monkey (Macaca fascicularis)."
http://www.biomedsearch.com/nih/Bartonella-quintana-in-cynomolgus-monkey/16485482.html

On Fig 2c:" erythrocyte with vacuole-enclosed suspect organism."

http://images.biomedsearch.com/16485482/03-0045-F2.jpg?AWSAccessKeyId=AKIAIBOKHYOLP4MBMRGQ&Expires=1491004800&Signature=qb870w8D5nZO%2B%2BR9gGzP5F7gNPY%3D

Fig 1a: Also looks like it has a vacuole & an organisms within the vacuole & within the rbc..you have to zoom in really hard to see the discoloration the vacuole produces.

http://images.biomedsearch.com/16485482/03-0045-F1.jpg?AWSAccessKeyId=AKIAIBOKHYOLP4MBMRGQ&Expires=1491004800&Signature=vbKeBuuCePx%2F79VtR528R0478YU%3D


I tried to zoom & expand it for you here:
http://s1275.photobucket.com/user/Jack_Dupre/media/Fig%201a%20copy_zpsp0u1duhi.jpg.html?sort=3&o=0


______________________________________
3) " Unusual trafficking pattern of Bartonella henselae -containing vacuoles in macrophages and endothelial cells"
" Bartonella henselae, the agent of cat-scratch disease and vasculoproliferative disorders in humans, is a fastidious facultative intracellular pathogen, whose interaction with macrophages and endothelial cells (ECs) is crucial in the pathogenesis of these diseases. However, little is known about the subcellular compartment in which B. henselae resides."

http://onlinelibrary.wiley.com/doi/10.1111/j.1462-5822.2005.00531.x/full

Bart in vacuoles pic:
http://onlinelibrary.wiley.com/store/10.1111/j.1462-5822.2005.00531.x/asset/image_n/CMI_531_f1.gif?v=1&t=j0wq739b&s=57ceeb657eea7c0f705960173fb7f19a84cd4ad4

" 2 or 24 h after infection, revealed that all internalized bacteria were within membrane-bound compartments"
http://onlinelibrary.wiley.com/store/10.1111/j.1462-5822.2005.00531.x/asset/image_n/CMI_531_f2.gif?v=1&t=j0wq739b&s=8a72e0d8ae65bb52a38a197c2d5c2c3f912f9f25


______________________________________
4) " Common infection strategy of the bartonellae. The drawing illustrates the general concept of reservoir host infections with Bartonella. Following transmission by an arthropod vector (a), the bartonellae colonize the primary niche, which probably involves entry into migratory cells (b) and transport to the vascular endothelium (c), where the bacteria persist intracellularly. From the primary niche, the bacteria are seeded into the bloodstream (d), where they invade erythrocytes and reinfect the primary niche. After limited replication inside the red blood cell (e), they persist in the intraerythrocytic niche (f) competent for transmission by a bloodsucking arthropod (g)."

http://cmr.asm.org/content/25/1/42.full

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______________________________________
5) " Some species are able to infect erythrocytes while others simply attach to host cells"

http://www.microbiologybook.org/mayer/ricketsia.htm

" FIG 5 Entry of Bartonella into human erythrocytes. The scanning electron micrographs show B. bacilliformis first inducing indentations in the erythrocyte surface (left) and then invading at the resulting pits (right). Note that neither the deformation of erythrocyte membranes nor the actual invasion process appears to involve rupturing of the erythrocyte surface. (Adapted from reference 31 with permission.) "


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" The uptake of single bacteria or small conglomerates in the zipper-like mechanism yields Bartonella-containing vacuoles (BCVs) that fail to acidify and to fuse with lysosomes but instead accumulate in the perinuclear space (246). Residence in perinuclear vacuoles was also reported for B. bacilliformis (429) and B. quintana (60, 395), indicating that this mode of intracellular persistence may be a common property of the bartonellae and possibly reflect a cellular niche colonized during infection in vivo."
" Intraerythrocytic persistence.It has not been resolved in great detail how erythrocyte infection proceeds after the invasion process, e.g., in what way the bacteria gain access to the nutrients inside the red blood cell. B. bacilliformis was reported to end up in intraerythrocytic vacuoles when the invaginations from membrane deformation bud off during “forced endocytosis.” However, the bacteria also occasionally appeared in the lumen of infected erythrocytes in vitro (31), but it is not clear if the bartonellae are able to actively leave the vacuole under physiological conditions. The examination of rat erythrocytes from in vivo infection with B. tribocorum indicated that the bacteria remain inside a membrane-bound compartment (394).
Experiments on the rat model also indicated that red blood cells would be initially infected by not more than one or two bacteria, which divide two or three times, giving rise to eight bacteria per erythrocyte on average, and then persist for the residual life span of the red blood cell, which was apparently not shortened by infection (394)."

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Lymedin2010
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TNT, thought you might like this.

" Blood smear showing a typical Bartonella-like bacteria (office lab)

These images are from my office lab which now performs in house, diagnostic blood smear examinations for our patients. All of these images come from patients with negative serological blood test results.

This a Giemsa stain from my office lab. The tiny dot at the bottom of the slide is typical of Bartonella-like organisms. These are typically very small. The bacteria stains dark blue. Note, the larger purplish splotch indenting a red blood cell comes from platelets. The larger cell is a typical white blood cell called by various names, including: neutrophil, poly, granulocyte. "


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http://lymemd.blogspot.com/2013/09/blood-smear-showing-typical-bartonella.html

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TNT
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quote:
Originally posted by Lymedin2010:
TNT, thought you might like this.

" Blood smear showing a typical Bartonella-like bacteria (office lab)

These images are from my office lab which now performs in house, diagnostic blood smear examinations for our patients. All of these images come from patients with negative serological blood test results.

This a Giemsa stain from my office lab. The tiny dot at the bottom of the slide is typical of Bartonella-like organisms. These are typically very small. The bacteria stains dark blue. Note, the larger purplish splotch indenting a red blood cell comes from platelets. The larger cell is a typical white blood cell called by various names, including: neutrophil, poly, granulocyte. "


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http://lymemd.blogspot.com/2013/09/blood-smear-showing-typical-bartonella.html

Yep, that's exactly what I see in my smears.
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mustardseed2
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Possible Babesia in the middle there?
It was much more pronounced through the lens; my cell phone doesn't take the most amazing images.

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I've done dozens of stains, and this is one of the only halo structures I've seen. I figured if I have Babesia, I would have seen more by now. Maybe I don't have Babesia in my fingers [Razz] . Starting Mepron this weekend anyway.


I took a shot of this stuff because it stained much more vivid than than anything else I've seen before. Again, not a great quality image, but it was made up of multiple little dots and rods. Definitely not platelets. Check out that inclusion in the RBC to the left. Not sure I've seen anything like that before:

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Lastly, I'm still not 100% sure what these dark staining specs are inside of my WBCs:

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Hope everyone's having a good weekend!

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rainboworiver
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Hello everyone, I am here to report that my blood is getting better and better. However, I do want to know what the clusters are in the video. The first cluster consists of black dots. The second one is different from the first one.

https://vimeo.com/211115748

Thanks!

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rainboworiver
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One key thing for my feeling better and better is Kelp. I take 6-10 capsules a day. Also every time I ate raw garlic, I herx.

I tried hyperbaric oxygen twice. I tried bee venom twice,but allergic to it.

Hope it helps.

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TNT
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quote:
Originally posted by rainboworiver:
Hello everyone, I am here to report that my blood is getting better and better. However, I do want to know what the clusters are in the video. The first cluster consists of black dots. The second one is different from the first one.

https://vimeo.com/211115748

Thanks!

I'm glad your blood is looking better and better! What are the elements of your protocol at the present besides kelp and garlic? Too bad you're allergic to bee stings. That's a bummer. Did you have trouble breathing? A reaction is not the same as being allergic. How did you react to hyperbaric oxygen?

The clumps of black dots (at :15 in your video) are clumps I've seen in my blood, too. I have not figured it out, though I think it is probably a collection of cellular debris of some kind or another. I've considered a collection of lipids, too, except lipids should be nearly perfectly-round objects with a different light refraction.

But, the other "clumps" you are referring to (1:00 in your video) are cellular debris and remnants from white blood cells. That I am certain of.

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rainboworiver
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Lymedin2010, if platelete doesn't have DNA, why does it stain purple? I thought only DNA stains.

Also could someone give me some pointers on where to buy and how to get started with staining?

Thanks!

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rainboworiver
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TNT, to answer your question, I am recapping from my previous posts and now adding new stuff. Here you go.

On Feb 23, I stopped all abx, my blood was loaded with organisms. I started the following herbs: Baical skullcap, 2 dropfuls 3x a day, kelp (2 tablets a day), agrisept 20 drops 3x a day. I did experience herx (not very severe). This is in addition to serrapeptase, Japanese knotweed, nattokinase, protease and probiotics that I have been taking for over one month. Additionally, I have been doing IV EDTA chelation plus vitamin c and b and glutathione once every 1 to 2 weeks for a couple of months. I just finished the last of the 10 EDTA sessions. I believe that EDTA and the enzymes contributed significantly to breaking up the biofilm clusters and red blood cell clustering (the rouleaux) problem. I significantly restricted sugar intake (fruits and startch) for two weeks, but couldn’t last long, so I have relaxed about eating a few of fruits. A few days ago, I added pinella 10 drops twice a day and coptis one dropful twice a day. I also added lauricidin, two teaspoons a day. I experienced herx again (not too severe).

Two weeks ago, I tried HBOT at 2 ATA for 90 minutes. I felt great the first time. I didn’t feel the same drastic effect after the 2nd time. I had to stop to wait to get my doctor’s clearance on brain aneurysm.
I tried bee venom therapy, the area blew up to the size of an orange. I stopped after two tries. I may try again in a couple of weeks.

One week ago, I added biotherapy karlovy vary thermal spring salt, it alkalines the blood and adds back the minerals I lost during EDTA chelation. Two weeks ago, I started colonic hydrotherapy, once a week. At the end of the 7th session, they will restore colon flora with probiotics. I have been feeling better after each colonic.

I have been using self-made LED lights (red, yellow and infrared) a couple of times a week for 30 min to 60 mins. I don’t know if this helped with lyme, but didn’t notice any reaction afterwards. It did work magic on the bee sting (orange size) in stopping the unbearable itch within 10 minutes, and calming down the angry redness and reducing sewelling.

I blend lots of garlic in roasted eggplant/pepper, each time I herx from it. I am sticking to this though I probably reek garlic from afar.

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Lymedin2010
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Clean stains! Both look like it could be babesia. The 2nd one almost looks like a lopsided maltese cross, although it did not stain purple/blue.


Sometimes I do see other babs stains that look slightly pinkish though...you be the judge.


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_______________________________
The rbc's stain pink & platelets should stain violet but can also look pinkish if the platelet granules are dispersed and if they are concentrated then slightly more violet or pretty close to purple & wbc's nucleus blue (which may look purplish) & leukocyte chromatin magenta. Bacteria should come out blue & sometimes look purplish as well. All this can also vary upon magnification & your lighting & the camera that picks up the color. Stains are best viewed under 100x oil.

Mustarseed uses this one below & I tried it too...it is great & relatively cheap. You just make the peripheral blood smear just like in this video. You spread & fan out the blood to make a nice corona & then let it air dry in a closed environment (to avoid contamination) for 10-15 min & then stain.

https://www.youtube.com/watch?v=4NkgEjPKzBA


Then you just dip a few times in each container & blot between each container so as to avoid excess drip into the adjacent container (touch the edge of the slide to a tissue to remove excess stain solutions). Mustardseed gave instructions to us a few weeks ago, in 3 or so pages back, as he has managed to perfect the technique.


http://www.ebay.com/itm/Vet-Supply-Dip-Quick-Stain-Refill-Kit-Testing-Labs-Slides-Vet-180-ml-/351928491512?hash=item51f0929df8:g:gMEAAOSwA3dYSaz~


You can buy refills here:
http://www.ebay.com/itm/Vet-Supply-Dip-Quick-Stain-Refill-Kit-Testing-Labs-Slides-Vet-500-ml-/351527003416?hash=item51d8a46518:g:wNIAAOSwa~BYQYQT


Great that your blood has less spiros. Another doctor reports that the levels of their blood & their patients blood can go up & down from time to time, as this person has checked for a few years now. So the levels can wax & wane.

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Lymedin2010
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So tough to say really for sure on the babs stains, but it could be. I am not totally satisfied & don't get that content feeling for either of the images honestly...but they could just as well be.

I would just keep hunting for more evidence.

I have a few what look like babs stains on mine too, but I am not happy with them & dismiss them so far.

Pic 1 issues, no babs organism on the edge & just a plain circular halo.

Pic 2: The white around what almost look like a maltese cross & lack of blue/purple stain. I can forgive the stain, but that white surrounding I wonder & bothers me.

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Lymedin2010
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I rewrote this for everyone....


CHEAP & ACCESSIBLE Giemsa/Wright type stain:
You just make the peripheral blood smear just like in this video. You spread & fan out the blood to make a nice corona & then let it air dry in a closed environment (to avoid contamination) for 10-15 min & then stain.

https://www.youtube.com/watch?v=4NkgEjPKzBA

Here is a lab example of Diff Quick/Dip Quick stain procedure. I would not use tap water as they did here, but RODI or distilled water instead.
https://www.youtube.com/watch?v=V9nFq8BvW-Q


1) To stain you just dip in container #1 a few times they say 5. So 5x times up & down to dip. We found that we get better results if we dip 10 times over 5-7 seconds. Then blot dry (touch the edge of the slide to a tissue to remove excess stain solutions)

2) Then don't waste time & go straight to dipping in container #2, it says for 5 dips/seconds. But again 10x dips over 5-7 sec worked better for us. Then blot dry.

3) Quickly into 3rd container, we found best 7 dips over 3-5 sec. Then REALLY QUICKLY rinse it with RODI or distilled water right away. A 5 sec wait between the water rinse & the last dip of #3 might mess it up, so rinse it quickly but GENTLY.

The dipping times are suggested, but there is some free play & you can try and see what works best for you. Some people even try alternative means, for instance instead of air drying the blood sample, they bake it in an oven @100F.


The Dip Quick stain kit here:

http://www.ebay.com/itm/Vet-Supply-Dip-Quick-Stain-Refill-Kit-Testing-Labs-Slides-Vet-180-ml-/351928491512?hash=item51f0929df8:g:gMEAAOSwA3dYSaz~


You can buy refills here:
http://www.ebay.com/itm/Vet-Supply-Dip-Quick-Stain-Refill-Kit-Testing-Labs-Slides-Vet-500-ml-/351527003416?hash=item51d8a46518:g:wNIAAOSwa~BYQYQT

_____________________________________
Will the stain containers have residue after many uses?
After many, many uses yes. Most of it will settle to the bottom & so over time try not to shake or disturb the containers as much as possible. If you want an efficient way of managing your stains, then get a Wheaton glass Coplin staining jar with a lid. You can then use this a few times & discard for new solution when you deem necessary.

http://www.ebay.com/sch/i.html?_from=R40&_trksid=m570.l1313&_nkw=Wheaton+glass+Coplin+staining+jar&_sacat=0


You can also use these cheap microscope slide holders, but the lid on these is not air/liquid tight & you may have to store them in durable plastic bags & away very safely or just discard the solution after doing multiple slides at one time.

http://www.ebay.com/itm/50pcs-Microscope-Slide-Mailer-Case-Box-Plastic-hold-5-slides-/181675192796?hash=item2a4caf71dc:g:IB4AAOSwU9xUNiBb


Another type of staining container:

http://www.ebay.com/itm/White-Acid-alkali-PP-Plastic-Slide-Staining-Jar-Stained-Frame-Set-Lab-Supplies-/390583411727?hash=item5af095e40f:g:ysgAAMXQKLdR28Kc


Droppers to use with other staining method, the Diff Quick & Dip Quick (Vet version) do not require the droppers since you just DIP the slides in each jars successively.

http://www.ebay.com/itm/100-PCS-3ml-Disposable-Polyethylene-Eye-Dropper-Set-Transfer-Graduated-Pipettes-/262882957820?hash=item3d350b99fc:g:9K0AAOSw3YNXZhLU


***Anyone staining should know that the fumes from these stains is toxic & you should stain in a well vented & open area, although the staining videos that I have seen appear not to have taken those precautions. With Lyme Disease I would not risk exposure to any toxic fumes.
_____________________________________
For Giemsa/Wright stain in general:

Structure Colour
Erythrocytes Pink/yellowish red
Platelets Violet/purple granules
Neutrophils Blue nucleus, pink cytoplasm, violet granules
Eosinophils Blue nucleus, blue cytoplasm, red granules
Basophils Purple/dark blue nucleus, violet granules
Monocyte Violet nucleus, light blue cytoplasm
Bacteria Blue
Spermatozoa Pale blue in the acrosomal region and dark blue in the post-acrosomal region

https://en.wikipedia.org/wiki/Diff-Quik
_____________________________________
How do I know what is an organism & what is human component in the stain?

From the color chart above & you need to view & study the known examples of the various stained pathogens. If you click on the source pictures below sometimes there is description of the stain used & other times you can click on "view page" to see the source page that it came from. You will probably encounter more stained subjects that are artifacts or questionable then to get actual hits that match the known stained pictures. The better you get at staining the easier it becomes, but always expect to discard stained subjects more than you identify.

BABESIA, search "babesia stain":
https://images.search.yahoo.com/search/images;_ylt=AwrBT8Z7CuFYEL0AcihXNyoA;_ylu=X3oDMTEyZ3Z1b2hvBGNvbG8DYmYxBHBvcwMxBHZ0aWQDVUkyQzNfMQRzZWMDc2M-?p=babesia+stain&fr=chr-greentree_f f

BARTONELLA, search "bartonella stain":
https://images.search.yahoo.com/search/images;_ylt=A0LEV1s0CuFY7ZoAAIFXNyoA;_ylu=X3oDMTEyZ3Z1b2hvBGNvbG8DYmYxBHBvcwMxBHZ0aWQDVUkyQzNfMQRzZWMDc2M-?p=bartonella+stain&fr=chr-greentre e_ff

ANAPLASMA, search "anaplasma stain":
https://images.search.yahoo.com/search/images;_ylt=A0LEVyRHC.FY6QUAvwdXNyoA;_ylu=X3oDMTEyZ3Z1b2hvBGNvbG8DYmYxBHBvcwMxBHZ0aWQDVUkyQzNfMQRzZWMDc2M-?p=anaplasma+stain&fr=yfp-t

ERLICHIA, search "erlichia stain":
https://images.search.yahoo.com/search/images;_ylt=AwrBT9OwC.FY5E0AFVNXNyoA;_ylu=X3oDMTEyZ3Z1b2hvBGNvbG8DYmYxBHBvcwMxBHZ0aWQDVUkyQzNfMQRzZWMDc2M-?p=ehrlichia+stain&fr=yfp-t

"borrelia stain"....check what stain was used, there are many different stains for Borrelia:
https://images.search.yahoo.com/search/images;_ylt=A0LEVwxjDeFY7EUAABVXNyoA;_ylu=X3oDMTEyZ3Z1b2hvBGNvbG8DYmYxBHBvcwMxBHZ0aWQDVUkyQzNfMQRzZWMDc2M-?p=Borrelia+stain&fr=yfp-t


You can do many other searches, such as "Babesia microti stain" or "babesia wright stain" or "babesia giemsa stain"...etc.

[ 06-14-2017, 02:27 AM: Message edited by: Lymedin2010 ]

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rainboworiver
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Lymedin2010, thank you so much for taking the time to write up the how-to guide to staining. I had a mental block about starting, now I feel I can tackle it. [Smile] I know I have organisms in my RBC. How does one know if it is lyme, babesia or other protozoa in RBC?

I guess before my blood was loaded with so many different organisms, it didn't matter what they were. Now that I have gotten the load way down, I am getting more curious about what else is out there. With the treatment I did in the last month, I have made more progress in one month both in terms of live blood and symptoms than the last 8 years.

Are those pictures from your blood? they are fantastic pictures.

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Lymedin2010
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rainbow, I answered your question & gave some more important updated info above.

Those are not my samples, but from the links above.

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Lymedin2010
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"Detection and treatment of babesiosis are important tools to control babesiosis. Microscopy detection methods are still the cheapest and fastest methods used to identify Babesia parasites although their sensitivity and specificity are limited. Newer immunological methods are being developed and they offer faster, more sensitive and more specific options to conventional methods, although the direct immunological diagnoses of parasite antigens in host tissues are still missing. Detection methods based on nucleic acid identification and their amplification are the most sensitive and reliable techniques available today; they are fast, very specific and although most of them relay on sophisticated equipment, new methodologies are being developed without the need of expensive apparatus. Importantly, most of those methodologies were developed before the genomics and bioinformatics era, which leaves ample room for optimization. For years, babesiosis treatment has been based on the use of very few drugs like imidocarb or diminazene aceturate. Recently, several pharmacological compounds were developed and evaluated, offering new options to control the disease. With the complete sequence of the Babesia bovis genome and the B. bigemina genome project in progress [4], the post-genomic era brings a new light on the development of diagnosis methods and new chemotherapy targets."


 -

"Babesia especies in various hosts and tissues. A) Babesia bigemina in bovine erythrocytes. Blood smear stained with Giemsa. B) Babesia bovis in bovine erythrocytes. Blood smear stained with Giemsa. C) Babesia microti in mouse erytrocytes. Blood smear stained with Giemsa. D) Babesia bigemina kinetes in Rhipicephalus (Boophilus) microplus haemolymph. Haemolymph smear stained with Giemsa. E) Babesia bovis in a bovine brain capillar. Histological section of brain tissue stained with Giemsa. F) Detection of antibodies against Babesia bigemina by the Indirect Fluorescent Antibody Test (IFAT). Bovine antibodies were detected by a secondary, donkey IgG anti- bovine IgG bound to Alexa-Fluor 488. Images were obtained with an objective of 100X."


https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3355466/

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mustardseed2
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Ya that inclusion in my second picture is really weird. Sort of like some of those pics you posted.

I'm feeling like absolute garbage today, so maybe I'll dive in and try a few more stains.

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Lymedin2010
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Staining seemed so easy & straightforward when presented by others, we did not realize there will be this much hunting involved & questioning and second guessing.


Personally, I find microscopy with this horrible disease therapeutic. Rather than just giving into it, we are doing something & trying to figure things out & better than the alternative of doing nothing at all. JMHO.

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Lymedin2010
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At time 9:24 & onward she says that her doc thinks she has MS. She is on treatment for Lyme Disease.

https://youtu.be/VN_W9K5Wt_0?t=564

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thelowdown
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Hello friends, I've been ill for around 15 years and making steady progress toward recovery over the past three years with detoxification. I had mercury and record-breaking copper toxicity along with mold exposure and chronic inflammatory response syndrome. Now my doctor says I also have Lyme disease so I borrowed a microscope from a friend yesterday. My blood looks awful (massive rouleaux) but I could not see any spirochetes.

I did see a number of red blood cells that were twitching and maybe revolving. They seemed to have some darker curves which would twist and untwist but I'm still unsure as to what I was seeing. I understand that it may take 24 hours or more for spirochetes to emerge, but after 16 hours or so, my slide looks all dried up. Am I doing something wrong? How do I keep it hydrated? My procedure was to place a drop of blood on the slide and then put a slipcover on top.

I should add too that Charles Barker adds pure copper powder to the blood to force the spirochetes right out, but I don't have pure copper. Anyone else experimented with other ways of forcing them out quickly?

--------------------
Eric
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Lymedin2010
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What magnification are you looking at them?

If you use 100x oil, then add oil immersion to the slip cover & that will seal them & avoid dehydration. If you are not using an oil objective then you can take a Q-tip & pull the tip out to a point & dip it in Vaseline & apply it to the edges of the cover slip only to seal it.

https://www.youtube.com/watch?v=IZ1scQdbmtg&list=PLrV8FYOIQcamUAh2BjIPVcKI-mJ-zaigc

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thelowdown
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Thank you lymedin2010!! That was very very helpful. With your tip I've been able to keep my blood hydrated with no sign of deterioration going on 24 hours now.

I have not seen any spirochetes, but there are tiny little critters running around that clearly do not belong there. As for size, you could fit a couple hundred of them in a red blood cell. On another YouTube video I saw something similar being described as parasites.

There is also something strange I would love to have your feedback on. I have a number of cells that look to be the same size as red blood cells that twinkle. It's almost as if I had a red blood cell that was full of the little parasite thingies. The twinkling might just be motion of the bright dots.

Here's a picture of one of my disco balls.
https://www.screencast.com/t/ecJxO8rdc8

Unfortunately when I try to capture video, my software crashes. I'm thinking of getting a better camera and maybe microscope also because sure would be nice to zoom in on this stuff...

--------------------
Eric
http://howirecovered.com

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Lymedin2010
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That looks like a white blood cell with granules inside the cytoplasm.

I have tested a few cameras so far & out of Canon 40D, Sony a6300 (HD), Sony FDR-X1000V, & Panasonic G7 (4K), so far the Samsung Galaxy S5 has the best interpretation of lighting (WB) & produces the best video in 4K & the Sony FDR-X1000V comes in 2nd. The Sony is a nightmare in terms of battery life, ease of access. The Samsung phone you can also send video out to a TV via HDMI & or sync with a larger tablet for viewing. If you use the Sync method you can also charge it via USB & have PC access to files instantly while recording & it really works out great. There are time lapse apps on the Google stores that you can use as well.

I show you some of the ways to hookup a phone in this MY MICROSCOPE FaceBook group here:

https://www.facebook.com/groups/MyMicroscope/permalink/1648008998815428/

______________________________________
The ability of the spirochetes to drill forward and instantly backwards is a unique feature of spirochetes. Here is a lab reference B burgdorferi B31 strain that shows this inherent feature at time 26s, but the whole video is worth a watch. Keep this in mind when you watch some of my tick videos & my future tick video that will show spirochetes in a new light.

https://youtu.be/RRfHiweJ8yY?t=26


Here is another great Bb B31 video from this same person.

https://www.youtube.com/watch?v=3HRVGTM57LE

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thelowdown
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Ah, i see, a lymphocyte, thank you.

Never occured to me I could use a DSLR with the microscope? Do you like that better than the purpose built microscope cameras? I do have a G7 here.

Does everyone have those little white jitterbug's? wondering if those are normal or parasites... I have seen some that are a little larger than others.

That video is really interesting - Those spiros can move!

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Eric
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birthdaysuit
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This thread is truly eye opening and scary.
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thelowdown
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i'm pretty sure it's babesia i'm looking at in my blood and my daughter's. just ordered a better omax camera and will put up videos eventually.

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Eric
http://howirecovered.com

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bluelyme
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yes birthday yes,
lowdown -i have seen those lil white jitterbugs in a lot of peoples samples some say normal flora other say baby ketes or other pathogens.some one had a name for them do you remember, mustard?

here is a vid from our french counterparts showing a quick encyst on 2nd part , my friend translated and said it says that 2 spirochetes are being birthed from the round form
https://www.youtube.com/watch?v=d3dHFBiLac4

--------------------
Blue

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thelowdown
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wow, bluelyme - that french video is amazing. i think i see lots of balled up spirochetes in my blood but i haven't seen it happen.

I started making videos and posted my lyme story with vids here: http://howirecovered.com/lyme-disease-and-co-infections/

For now, the resolution in my videos is in great, but I am getting a new microscope this week!

--------------------
Eric
http://howirecovered.com

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TNT
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quote:
Originally posted by thelowdown:

There is also something strange I would love to have your feedback on. I have a number of cells that look to be the same size as red blood cells that twinkle. It's almost as if I had a red blood cell that was full of the little parasite thingies. The twinkling might just be motion of the bright dots.

Here's a picture of one of my disco balls.
https://www.screencast.com/t/ecJxO8rdc8


Lowdown & Blue..... as Lymedin said, those twinkling round cells are white blood cells. I can't quite discern from the picture which white cell it is, but I would lean towards a granulocyte such as basophil or eosinophil. Notice the coarse granules (lysosomes) in the cytoplasm.
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bluelyme
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I have seen those disco balls in a few peoples blood now .is it fungal?

--------------------
Blue

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TNT
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quote:
Originally posted by bluelyme:
I have seen those disco balls in a few peoples blood now .is it fungal?

They are, without a doubt, white blood cells!
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Lymedin2010
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WORLD PREMIER:

For the first time ever (that I know of) I capture time lapse of a SOP (String of Pearls) in squashed ticks, which the pearls then go on to develop into baby spirochete.

Subjects were filmed using various microscopes, objectives, lighting sources, &
different camera systems.

https://www.youtube.com/watch?v=7OaPWZBW36Q


**Use the PAUSE button for this video to read long texts, otherwise the video would have been even longer.

I also reveal PCR testing toward the end of the video.
This video starts out slow & gets better with more evidence
over time...clips of 400x, 630x, & 1000x.


________________________________________________
WHAT WE WILL SEE IN THIS VIDEO:


-What Tick juice looks like without any spirochetes.

-Tick juice with spirochetes.

-Rigid "Needle Form" spirochetes.

-Needle forms wake up to start motility.

-Spirochetes drill/spiral forward AND backwards.

-What stained BSK-H B burgdorferi grown spiros look like.

-Longer & typical spirochetes in ticks.

-Spiros that split into 2, 3, 4...daughter cells.

-Spiros that drill/spiral erratically.

-Spiros that just gyrate and stagnate similarly to Lyme Diseased blood.

-Granular forms (blebs), spiros developing from granules.

-Very tiny spiros, medium, & larger ones, thin & thick ones too...many morphologies.

-Early biofilm formation & aggregation of spiros.

-SOP (String of Pearls) & segmented spiros & spiros w/light & dark bands/spotting.

-Time lapse breakup of SOP & blebs/segments grow into baby spiros.



________________________________________________

STAGES IN SPIROCHETE PRESENTATION FROM TICK JUICE:


1) RODI or Distilled water, or other solutions can cause spirochetes to form cysts.
Sometimes it can take an hour or more for them to come out of cyst morphology,
depending on the solution. Water is just one way to force them to cyst up.


2) Spiros uncyst & can be very rigid and in "Needle Form." This seems to be a
shock stage, where they are perhaps still adjusting to osmosis of the added solution.

3) When nutrients are present they can move VERY rapidly & go through rapid splitting once uncysting & adjusting. These are VERY FAST growers & nutrients dictates they produce & split VERY rapidly for these spiros. In this stage many 2 daughter spiros will be seen seperated by a thin bit in between the daughters.
Sometimes 3, 4, & more daughters can be seen.

4) When they are left in nutrients for longer periods they tend to grow thicker and longer. Starved nymph ticks may not present with longer or thicker forms of this particular spiro, they tend to remain as smaller units in low nutrients.

5) When nutrients are depleted the spirochetes have less motility or simply gyrate in place in stagnant forms, sometimes very similarly to what we have seen in Lyme Diseased blood. The solution they are in & nutrient availability dictate to some degree how the spirochete evolves and moves.


6) Even in stagnant forms & in forms much smaller than the full blown typical BSK-H grown form, they tend to coalesce, secrete a cloudy/milky substance into the solution, & start forming a slimy biofilm. The cloudiness in the solution over time obscures the visibility of the spiros.

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Lymedin2010
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40x * Zeiss 15x eyepiece = 600x + cam zoom on a Samsung Galaxy S5 with this microscope adapter. The Zeiss optics are top notch when resolution matters!

https://www.youtube.com/watch?v=PwzCiPgYeDw

http://www.ebay.com/itm/Universal-Cell-Phone-Adapter-Mount-For-Spotting-Scope-Telescope-Microscope-/222456604733?hash=item33cb72683d:g:b8cAAOSwmgJY3K2M


Using this light bulb on a standard light fixture right under the microscope condenser:
 -

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TNT
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Lymedin, you are the KING OF SPIROCHETES!!! Great research and a wonderful video production!!! I don't know how a person can improve on that proof. The time-lapse at the end with the string of pearls budding & molting into juvenile spirochetes was just spectacular! And to see them swim away....!!! Incredible!

It would be nice to see similar time-lapse of Bb SOP molting into juvenile spirochetes. Regardless, your documentation of this process is truly remarkable even though they are not Bb spirochetes!

This is cutting-edge research if you ask me! Way to Go!!!

I've got some potential evidence of invasomes that I plan to share before long. Ha Ha, you didn't think you would hear that from me did you? I'm still a bit hesitant to say they are "Bart" invasomes just because there's no way to prove what organisms they are. But, the vacuoles are definitely visible and the organisms inside are unmistakable.

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Lymedin2010
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Thanks TNT.

Refinements will produce even better video on the next round.

I am eagerly awaiting your fluoro & bart videos and might even get inspired to get a fluoro scope as well.

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mustardseed2
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Terrible quality, as per usual with my cell phone camera, but here's a possible Bartonella-like organism. It's hard to tell from this picture, but it's pretty rare something gets colored blue in my stains. Edit: not sure why the image is so small. You'll just have to take my word that there's something there [Razz]

 -

Anyone see stuff like this next image? I see it quite often. I'm not 100% sure, but I think it's the leftovers of an exploded white blood cell.

 -

Lastly, still getting these dark purple spots inside of my WBC nuclei. This one is particularly big. They never show up outside of the nucleus, and it's possible that it's an artifact. I really don't know.

 -

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TNT
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quote:
Originally posted by mustardseed2:
Terrible quality, as per usual with my cell phone camera, but here's a possible Bartonella-like organism. It's hard to tell from this picture, but it's pretty rare something gets colored blue in my stains. Edit: not sure why the image is so small. You'll just have to take my word that there's something there [Razz]

 -

Anyone see stuff like this next image? I see it quite often. I'm not 100% sure, but I think it's the leftovers of an exploded white blood cell.

 -

Lastly, still getting these dark purple spots inside of my WBC nuclei. This one is particularly big. They never show up outside of the nucleus, and it's possible that it's an artifact. I really don't know.

 -

Good work! That's a good possibility the first pic could be showing Bart. The two objects could also be other things such as Babesia merozoites, since they look more oblong than most Bartonella on the outside of a RBC. But it's a little hard to tell for sure.

I agree with you on the second pic. It's also referred to as a "smudge cell" in a Giemsa smear. I used to see lots of them too. The past few smears I've done I've seen very few. It seems to have coincided with the addition of Azithromycin (in addition to the other meds). I'm almost certain the smudge cells are a result of the Anaplasma infection, but Azithromycin is not very active against the "spotted" Rickettsias. Then again, the past few smears I've seen very few typical Bartonella on the peripheral of my RBCs. So, maybe the Azithro is hitting Bart which is causing, or at least contributing to, the smudge cells in my case.

As for your 3rd pic, my opinion is that what you are seeing is normal, except that it appears like the Neutrophil is superimposed on the RBCs. It appears ghosted too. This is a bit out of the ordinary. But, the clumpy, non-homogenous nucleus is very typical, and I can't really say that this particular cell looks abnormal (to me),except for what I mentioned.

I've been a bit bummed out lately. I haven't been doing as well and I haven't had the energy or focus to pursue things as I would like. It doesn't help that the past number of AO stains I've done have flopped. I've gotten one good AO dried smear since I've gotten the scope (and one trial wet mount that did ok). I don't know if it's the stain or something seemingly inconsequential I'm doing wrong that is causing my smears to wash (or soak) off. Believe me, I've tried varying my techniques, but no help. I think it's the stain, and will have to talk with the company to see if they can give me any clues as to why I had good success with the first dried smear, but none since. It's SO DISAPPOINTING!!! And, it's at a bad time for me, too. The AO dried smears could really give me some hard evidence concerning a few of these infections. My Giemsa stains are pretty definitive, but that absolute verification would really be helpful right now.

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mustardseed2
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Smudge cell... awesome! Thanks for putting any doubts I had about those to rest!

These smudge cells could very well be from Anaplasma. I tested positive for anaplasmosis through Igenex and have a moderate neutropenia right now. However, I've looked at hundreds of WBCs and have yet to see any anaplasma inclusions like the great pics you posted a page or two back.

Sorry to hear you haven't been feeling well. I don't know much about AO stains but they look pretty cool when I Google them.

What exactly are you looking for in them?

What infections are you treating right now?

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TNT
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quote:
Originally posted by mustardseed2:
Smudge cell... awesome! Thanks for putting any doubts I had about those to rest!

These smudge cells could very well be from Anaplasma. I tested positive for anaplasmosis through Igenex and have a moderate neutropenia right now. However, I've looked at hundreds of WBCs and have yet to see any anaplasma inclusions like the great pics you posted a page or two back.

Sorry to hear you haven't been feeling well. I don't know much about AO stains but they look pretty cool when I Google them.

What exactly are you looking for in them?

What infections are you treating right now?

Yeah, I would expect Anaplasma to cause the greatest number of smudge cells, but other infections such as Babesia and Bartonella can cause leukopenia as well. Even viruses can. You could try some Azithromycin with a tetracycline to see if that helps rectify your leukopenia. That is what has helped mine. The tetracycline without azithromycin did not do it for me.... which is odd, since effective treatment for Anaplasmosis is typically just Doxycycline. In resistant cases, Rifampin is added, but Azithromycin is usually not used or included.

Yeah, the fluoro stains are mesmerizing. I'm just looking for the things I typically see in my giemsa stains.....just confirmation really. It can be nearly impossible to differentiate between a platelet and a morula in a giemsa stain. Same thing with Babesia ring-forms. I have seen definite presence of both, but just want to see absolute confirmation with AO. The morulas are again the highest load, but just looking for that 100% confirmation.

I have (again) been treating Babs, but have (again) had to stop because of worsening Anaplasma and Lyme symptoms. The recent giemsa smear showed a higher load of morulas. A few months ago, before commencing Babs treatment, I essentially saw none. Interestingly, the last smear also began showing pretty clear evidence of Babs (ringforms). It seems the Malarone is bringing the Babs more to the surface, but allowing the Rickettsia & Bb to grow.

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mustardseed2
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Wow, super interesting. It seems that even though you stopped seeing the anaplasma, it hid undetected and then started multiplying when you stopped treating it. These infections are difficult.

The problem with Malarone and Mepron is that they don't mix well with Rifampin or Doxycycline, so it's difficult to treat Babesia at the same time as other coinfections.

Like I said, my Igenex came back positive for anaplasma at the same time my LLMD diagnosed me clinically with Babesia. Her idea was to do Zithro, Doxy, and Malarone all together. I definitely herxed on the Malarone so it was still working, despite being mixed with Doxy. Might want to try that if you haven't?

As for the neutropenia, mine was borderline, but stable, on the zithro/doxy combination, but when I added Malarone it rapidly plummeted. Exact same thing happened when I switched from Bartonella medications to Mepron. This article was interesting for me, you may have already seen it:

https://www.bcl-clinic.info/soubor/neutropenia-in-congenital-and-adult-babesiosis/

Best of luck with the new stains, I would love to see the results when you get it working.

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quote:
Originally posted by mustardseed2:
Like I said, my Igenex came back positive for anaplasma at the same time my LLMD diagnosed me clinically with Babesia. Her idea was to do Zithro, Doxy, and Malarone all together. I definitely herxed on the Malarone so it was still working, despite being mixed with Doxy. Might want to try that if you haven't?

As for the neutropenia, mine was borderline, but stable, on the zithro/doxy combination, but when I added Malarone it rapidly plummeted. Exact same thing happened when I switched from Bartonella medications to Mepron.

Thanks for the suggestion. I have actually done that combo, and it did seem to have been helping. The problem, though, seems to be when I get to the max dose of the Malarone there is not enough killing power for the Rickettsia or the Lyme. Because, this threshold seems particularly lower -- and coincides with -- the 4-week cycle of the Lyme. In other words, I do ok with full-dose Babs treatment until I hit the anticipated monthly cycle of the Lyme. Then things go south pretty quickly regarding my usual "Bart" (Anaplasma) and Lyme symptoms. Of course, as I said, this is evident in my blood smears, too. I'll probably have to maintain a lower dose of Malarone for longer than normal once I recommence.

That's very interesting that is what happened to you, too....at least according to the neutropenia. It seems unlikely to me that the Babesia would have caused increased neutropenia when you were on the heavy Babs drugs. I tend to think your Anaplasma has not been sufficiently addressed even though you cannot find visual evidence of it in your smears.

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Fluorescent microscopy will be fascinating once I can delve into it more in depth....and get my smears to turn out.

Here is a pic that Lymedin posted earlier:

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And, here is a page on Acridine Orange staining from Counselling Me Microscopy that shows how it is possible to see bacteria (probable borrelia) inside RBCs even when it is impossible to see with darkfield (in live blood):

http://counsellingme.com/microscopy/AcridineOrange.html

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Wow those bacteria really pop in that stain! Man I hope you can get it working, it looks amazing.

What was your maximum Malarone dose, out of curiosity? LLMDs are prescribing increasingly high amount of atovaquone for Babesia, because the old 750mg (1 tsp Mepron) twice a day just isn't cutting it for people.

For instance, my LLMD wanted to treat with 4500mg per day (3 tsp twice a day). We settled on 3000mg to not freak out the insurance company. 4500mg would be 18 Malarone tablets per day (and remember that the atovaquone tablet is relatively poorly absorbed vs the liquid). I get what you're saying though, you've got to keep on top of the anaplasma too, especially if it's popping up in your stains more frequently.

Of topic slightly, but is this YouTube video from someone in this thread?:

https://www.youtube.com/watch?v=oJgI3lr2ng8

The reason I ask is because it's labeled as Rickettsia. Do we know for sure that's what this is???
Because frig, I see these in pretty much every smear I do. Identical.

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quote:
Originally posted by mustardseed2:
Wow those bacteria really pop in that stain! Man I hope you can get it working, it looks amazing.

What was your maximum Malarone dose, out of curiosity? LLMDs are prescribing increasingly high amount of atovaquone for Babesia, because the old 750mg (1 tsp Mepron) twice a day just isn't cutting it for people.

For instance, my LLMD wanted to treat with 4500mg per day (3 tsp twice a day). We settled on 3000mg to not freak out the insurance company. 4500mg would be 18 Malarone tablets per day (and remember that the atovaquone tablet is relatively poorly absorbed vs the liquid). I get what you're saying though, you've got to keep on top of the anaplasma too, especially if it's popping up in your stains more frequently.

Of topic slightly, but is this YouTube video from someone in this thread?:

https://www.youtube.com/watch?v=oJgI3lr2ng8

The reason I ask is because it's labeled as Rickettsia. Do we know for sure that's what this is???
Because frig, I see these in pretty much every smear I do. Identical.

That's an incredible amount of atovaquone your doc wanted you on! I've only ever made it to 4 Malarone tabs daily. I've heard of the 3000mg daily, but not 4500mg! I'm surprised your insurance company would even cover the 3000mg!

Yeah, the morulas are more of an issue for me than Babs ringforms. Only since I started taking Malarone have I seen any probable ringforms in my blood. But, the morulas are definitely prominent.

That video is mine, ha ha. Like you, I've seen plenty of those in my blood! Besides spirochetes, bacilli are the most prominent pathogens in my live blood. I am assuming that these are Rickettsias since that is what I am finding in my stained dry smears. As an example, in the one successful AO dry stain I was able to get, I found 3 very distinct bacilli (rods) that exact size versus only one possible variant ringform.

It sounds to me like Rickettsias are your dominant pathogen as well.....

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Ya it might be. Figured 2 months of Doxy and 6 months Rifampin would have given me some relief.

Tomorrow when I have some time I'm going to start searching for rickettsia blood smears online.

I've seen pictures of morulae and I've seen pictures of thinner, smaller, ehrlichia species, but I haven't ever seen anything like these larger rods mentioned as rickettsial (outside of your video, which is amazing quality btw).

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Here is a sampling of my fluoro work. Sorry for the large size. I had to change my picture hosting site. Tinypic has become way too slow with too many glitches and possible security issues. I will continue to see if I can minimize the photos from my new site.


A bacillus that is probable Rickettsia:

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A closer and crisper pic of the same:

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A bacillus on or in a RBC:

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Another bacillus:

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Typical Bartonella on the peripheral of a RBC:

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A possible variant Babesia ringform (Take note of the platelets...they are a very pale green color and do not stain orange because they do not contain a nucleus with DNA or RNA. A Howell-Jolly body in a RBC will stain orange since HJ bodies are left-over nuclear material. But this is no Howell-Jolly body! HJ bodies are perfectly round):

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[ 05-10-2017, 11:07 PM: Message edited by: TNT ]

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Recent finding of Babesia ringforms in family member's blood:


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Closer pic of the same field:

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Ringform in a neutrophil (at the 2 o'clock position):

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mustardseed2
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TNT, your first post, the pictures aren't showing up.
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The Babs pics are, but the fluoro pics aren't??
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Ok, I just checked with a different device. Yes, the fluoro pics are not showing up. That's odd, because the pictures on both posts are from the same site, my google account. The giemsa pics were smaller format than the fluoro ones, but I don't see how that would make a difference.

Anyone have any ideas?

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mustardseed2
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Maybe there's a size limit per picture or something.
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Well, I just read this thread from start to finish for the first time. I'm finally caught up. I think the most interesting stuff was the debate about what those rod shaped things in TNT's blood are:

https://www.youtube.com/watch?v=oJgI3lr2ng8

Like I said, I'm finding the same stuff, and I think it was thatdudefromkansas who also found them.

Here's the trouble I'm running into: I can't find any credible information online that rickettsial bacteria are that large. The only thing that we seem to be basing this on is this video from CytoViva:

https://www.youtube.com/watch?v=cv_7I8D4KQc

...as well as the fact that TNT has a persistent HGA infection. However, as originally discussed several pages back, these thing also strongly resemble merozoites:

https://www.youtube.com/watch?v=FKEGZCZqZ_w

On top of this, as seen in the above video, merozoites often tend to remain attached to each other for some time. Sort of like this video by TNT:

https://www.youtube.com/watch?v=lOftWSU92fY

To complicate things even further, I suggest these might even be Borrelia round bodies (cysts). Check out this video:

https://www.youtube.com/watch?v=1HUtKungjvE

Notice not only the spirochete changing morphology, but also the two rod like organisms at the 9-10 o'clock position. I'm assuming those are round bodies as well.

All of this is really making my head spin. I'm not saying these rod/round things seen in blood samples is or isn't Rickettsial/Babesia/Borrelia, but what I am saying is it seems likely it could be any of those three.

Anyone know any professional microbiologist we can ask?

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TNT
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quote:
Originally posted by mustardseed2:


Here's the trouble I'm running into: I can't find any credible information online that rickettsial bacteria are that large. The only thing that we seem to be basing this on is this video from CytoViva:

https://www.youtube.com/watch?v=cv_7I8D4KQc

...as well as the fact that TNT has a persistent HGA infection.

The videos by CytoViva are authoritative, since their videos are with cultured and known organisms.

I struggled with the same questions, but finally had to go with the cumulative evidence (including treatment response), which for me all pointed to Rickettsias.

In the video by Muriel on the conversion to gemma cyst (the last video in your last post), that cyst looks like a rod bacteria only when it's oriented on it's side. Otherwise, you can tell it's a gemma cyst. The organism in my "Rickettsia" darkfield video was rod-shaped no matter what orientation it was.

I never thought about the rod-shaped, partly-dumbell-shaped organism in my phase contrast video resembling two attached merozoites. But, I agree, it could very well be.

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Ok, I posted those fluoro pics from tinypic, so now you can see them.
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Fluoro pics looking amazing.

Would be cool to put a guide together on how to do that.

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https://www.youtube.com/watch?v=lOftWSU92fY


quote:
Originally posted by TNT:


I never thought about the rod-shaped, partly-dumbell-shaped organism in my phase contrast video resembling two attached merozoites. But, I agree, it could very well be.

The organism could also be a bacillus (I'm using the term in it's general sense to describe a rod or coccus) bacteria in the process of binary fission....

Considering the overwhelming evidence I have of an Anaplasma infection, this possibility is more likely.

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Ya seems possible.

It could also be that the rods I see in my blood aren't the same as the rods in your blood, so I'm parlaying my skepticism towards my own sample onto yours. After reading through all that, I'm 99% sure my rods are these "gemma cysts". The color of the rods, the movement, the size and shape variation.

If you're not seeing that stuff, and you obviously see morulae, Rickettsial is probably a good guess.

In the long run, it doesn't really matter. The "best" treatment for HGA is also first line for Borreliosis (not that any of it is guaranteed to work).

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Yeah, I'm seeing plenty of morulas! Many of them are unmistakable since you can see the individual organisms inside.

Here they are, and I've got some exciting stuff for Lymedin coming tomorrow!!


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Lymedin, a couple pages back we had this in-depth discussion about Bartonella and invasomes. I felt there was more proof needed to validate what some were calling Bartonella vacuoles, or invasomes, in their live blood. I think that if we are seeing real Bartonella in vacuoles in our wet mounts we should be seeing them in our giemsa stains as well. Up til now I have not seen anything that resembled invasomes, but maybe I was just not noticing, or perhaps I was chalking them up to stain precipitate or artifacts.

I don't know if these are Prokaryotes or Eukaryotes--there is no way for me to tell-- so I'm not going to label them Bartonella. But, the clear presence of intracellular and extracellular vacuoles with multiple organisms inside is unmistakable in these pictures. You can clearly discern and even count the individual organisms within the vacuoles.

Here is what I have found. It is my blood:


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Notice that the invasome pictured in these next two pics is still fully encompassed within the red blood cell.

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[ 05-12-2017, 07:31 PM: Message edited by: TNT ]

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A couple more intracellular ones:


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This one may not be a good example since it is possible it's a platelet. Though, I'm pretty sure I can see two distinct organisms. It appears as though the organisms just shed the vacuole through the cell membrane.

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A few extracellular ones:

This first one is a bit unclear on the picture, but there are 4 organisms inside the vacuole. It appears to be on the outside perimeter of the red blood cell. The object beside it on the right I'm pretty sure is a platelet.

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And, here is one containing two organisms on the outside of a red blood cell:

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Wow! Someone just nailed a well-used, albeit decent, laboratory-grade American Optical 110 microscope (brightfield only) for $15.99 plus $35 shipping. A steal of a deal at that!

http://www.ebay.com/itm/122481498963?_trksid=p2055119.m1438.l2649&ssPageName=STRK%3AMEBIDX%3AIT&autorefresh=true


I love my AO 110 (fluoro) scope! It's a newer version of the 10/20 series. The only thing I caution concerning the 110 series is that accessories such as phase & darkfield condensers are a bit hard to come by and are usually fairly expensive. The 10/20 series scopes and accessories are still plentiful and cheap. I just saw a used oil darkfield condenser for the 110/120 series go for $120 (if I remember exactly), and phase contrast turret condensers are much more expensive and harder to find. As an example, the AO 10 series oil darkfield condensers by themselves (no case or funnelstop) can be had for less than $50.

And, from the limited experience I have (with the AO PLAN achromat phase lenses), the phase objectives and phase contrast condensers are not interchangeable between the 10 and 110 series scopes. The condensers will not mount interchangeably without some customization. The bigger problem, though, is that the annuli in the phase contrast condensers that fit the 10 series will not work with the newer PLAN achromat phase lenses that came with the 110 series. And vice a versa. I had bought a couple PLAN phase objectives (which came new on the 110/120 & 400 series only) and they would not work with the annuli in my 10 series phase contrast turret condenser. So, the annuli must be different in the phase condensers of the 10/20 series versus the 110/120 series scopes.

So, if you are in the market for a used scope and are looking at getting into different microscopic techniques, the AO 10 series scopes and accessories are going to be more plentiful and cheaper than the AO 110/120 & 400 series scopes. Just an FYI.

For brightfield, though, that scope I just linked to, was a steal!

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Great pics TNT!
Those vacuoles look very similar to a pic I posted here a few weeks ago, on the left. Great work!

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Way to go TNT.....that is some awesome work & findings!!!! Looks like you are the new resident co-infection go to man now. [Smile]

I am proud of you son!!!


It is great for us to discover & share with one another to make advancements & to learn further...we have all done some amazing stuff & contributed collectively!


My inclination for the clusters & vacuoles is Bart. As you have not found any evidence of Babesia in 2 & 4 pairs yet, with no maltese crosses, we can put Babesia on the back burner. If you look at the science papers & pictures that linked & some of the stains, notice that sometimes the stains show bacillus & other times some odd shape & what look like babesia deformed tear drops shape, or bean shape. I would imagine when the Bartonella is dividing it can present with various morphologies & appear odd and other than the bacillus shape. Just like with the spiro video that I put out, I think there will be various genetic expressions of Bart & some can be smaller & others larger overall as well, besides the mitosis growth stages.


From what I have seen with ricketssia, it is that they can appear as cocci, bacillus, & sometimes even longer as almost string-like. They also tend to grow in larger clusters & I would expect to see a much larger congregation of them in the same cell/spot. They too can form invasomes/phagosomes & can invade endothelial cells. From the evidence that I have seen it is one ricketssia per vacuole though, but I have not looked at all the species & rick's extensively as a whole. Maybe we can collectively spend a week researching rick's & we can post all the evidence and reanalyze. I will come back at some point & make remarks on the pictures individually.

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I started to write up my thoughts on Mustard's stains, but have to finish that too, but TNT hit it on the nail for the most part anyways.

I too have found similar structures when doing live blood preps, but I was never completely certain & never presented them. Then with the freezing method I was shocked & amazed at the congregation of them, while the wbc's & rbc's disintegrated largely. They also came up in my smears as well, although I wanted to do more stains & perfect them further before I made a true compilation. Now I have gotten them to POP out using fluorescence without even adding a dye yet, as there is a slight pinkish fluorescence they give off when using my Cree LED bulb, but it does not show up when using a halogen bulb though & so the light source has to be really bright & DAYLIGHT (no brown or yellow tinting) for them to be visible it seems. I want to be certain & do the actual fluorescent AO stain & do some stains on normal blood to make sure I am only presenting with these.


But I can put together a small preview video of them, so you see the pop in 4K. It is best when viewed by eye, then downgraded to 4K in terms of what it picks up, & then will probably be downgraded further on YouTube though...I will try to put it up tonight.

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Lymedin, I have found fairly certain evidence of Babs in my blood....pyriforms in a few RBCs to be exact. But no pairs or tetrads. But this was only in the last giemsa smear while I was on Malarone. So, I cannot rule out the possibility that some of what I show in those pictures are apicomplexans. I do think it interesting the difference of morphology between those pictures. They are definitely showing different species, and maybe different lifeforms. It's hard to tell just by looking at them.

Yes, Rickettsias can grow in large clusters, hence the development of morulas in Anaplasma & Erhlichia. Though, I had thought that with A. marginale, they presented as one or two organisms in infected erythrocytes. But, this electron micrograph would indicate that even those little red/purple dots on an erythrocyte is a vacuole containing multiple individual organisms. A. marginale organisms must be extremely small!

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Electron micrograph of a bovine erythrocyte infected with Anaplasma marginale (courtesy R.L. Sealock, Animal Parasitology Institute, ARS-USDA)


Just to clarify your comment about them being able to infect endothelial cells. Remember that Rickettsias aren't just capable of infecting endothelial cells, THAT IS THEIR PREFERENTIAL CELL AND TISSUE CHOICE!!! I don't know if they would survive if there was no endothelial --and to a lesser degree-- epithelial cells and tissues to infect. I don't think there would be any persisting infections anyhow. Of course, I don't think we would fare too well without those kinds of cells and tissues, either, ha ha! [shake]

So, DAD, when are you gonna show YOUR fluoro stuff? [Razz]

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I might be releasing a lessons video, so I will have to hold off. Trying to gather some more evidence.

I definitely have something very interesting that will give us a new twist & perspective on things though. I knew part of it as true, but there is new evidence that is eye opening & serves as a cautionary note to us as well.

TNT, have you tested again? Are you positive for Babs, Bart, Anaplasma, or Rick's...how long ago was your test?

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bluelyme
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Great work bros ! tnt lymed mustard any one heard from dude?
Here is from our french counterparts again .this footage is fascinating he shows a spirochete from canine saliva that is so much faster than any i have ever seen .also at the end it cyst"s and uncyst so quick really is a $hot.

https://youtu.be/diNuwoV6oGs

this applies to me because on some of my smears tnt said it appeared to be babesia canis and i remembered a few dog bites back in the day. This kete is scary quik may explain why some euro lyme go so neuro so fast .
may make think twice before getting kisses from your dog

--------------------
Blue

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TNT
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I have not been tested recently for anything. When I was tested, 3 Igenex FISH tests were negative, a Galaxy Bart test was slightly positive (read as a waning infection, ha), and HGE was negative through Igenex. A Brucella PCR was negative as well.

I don't have much faith in the tests. Microscopy and treatment response has been much more reliable.

An original Lyme test through Quest was negative. The original Bb WB through Igenex was CDC positive, the next one only two bands (read negative), and the most recent one was again CDC positive. So, go figure. This illustrates the reason for my lack of faith in tests. This also is the reason I don't believe the old party line that once you've been exposed to Lyme disease, you will test positive for a very long time-- perhaps the rest of your life -- long after the infection is gone. I personally don't think the antibodies last in the body nearly as long as we are told.

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TNT
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quote:
Originally posted by bluelyme:
Great work bros ! tnt lymed mustard any one heard from dude?
Here is from our french counterparts again .this footage is fascinating he shows a spirochete from canine saliva that is so much faster than any i have ever seen .also at the end it cyst"s and uncyst so quick really is a $hot.

https://youtu.be/diNuwoV6oGs

this applies to me because on some of my smears tnt said it appeared to be babesia canis and i remembered a few dog bites back in the day. This kete is scary quik may explain why some euro lyme go so neuro so fast .
may make think twice before getting kisses from your dog

Good find, Blue! And, NO, I won't be getting kisses from any dog!

Last year in the news was a warning from the CDC to wash your hands every time you've held and petted your cat. The concern was about getting Bartonellosis even if one was not scratched! All it took was a lick....in the face or on an open wound!

Here is one article I pulled up real quick from that time. It also talks about a woman dying from sepsis and multiple organ failure all as a result of a lick from her dog. So, Blue, you are right on!

http://www.dailymail.co.uk/health/article-3797101/Cuddling-kitten-KILL-CDC-warns-adorable-felines-carry-deadly-bacteria-causes-brain-swelling-heart-infection.html

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Lymedin2010
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TNT, thanks for the testing info.

Yea, I suggested to the FB group they check their dogs/cats tartar a few days ago & someone actually did just did. I had a gut feeling that we will find tons of spiros there. I also believe that one will find tons of spiros in wild animal mouths, including squirrels, deer, & mice.

I got bitten by a squirrel when I was young, as I thought they were cute & wanted to catch one...serves me right though! I also know someone who got bit by a squirrel a few years ago & she now has Parkinson's & is in a lot of pain with many Lyme symptoms. Her non-Igenex test showed negative though.

I just got a report from someone in the hospital that they just died recently of multiple organ failures after a tick bite & Lyme.

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Lymedin2010
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I hear you about the microscopy, I wish I did not waste my money on the tests, which I could have bought an awesome high-end microscope with all the testing money I spent.

Even if you test positive, can't tell if it was a past infection or current & you cannot keep testing & spending money on tests each 6-12 months.

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TNT
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quote:
Originally posted by Lymedin2010:
I hear you about the microscopy, I wish I did not waste my money on the tests, which I could have bought an awesome high-end microscope with all the testing money I spent.

Even if you test positive, can't tell if it was a past infection or current & you cannot keep testing & spending money on tests each 6-12 months.

AMEN! That's right.
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