Microscopy importance in veterinary medicine

[fastuno]

How important is a microscope in veterinary medicine for diagnosis of pathogens?

If a dog/cat owner comes in and tells you that their pet had been bitten by ticks & that they are just not acting like themselves, with malaise & perhaps joint pain/stiffness, do you attempt to diagnose by microscope?

Can the Borrelia burgdorferi spirochetes be visualized via microscopy?

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[Jamr0ckin]

Are you trying to do a homework assignment?

 

[fastuno]

No, I am just trying to determine if real world practice relies heavily on microscopic examination of live blood samples & in particular with Borrelia b. (Lyme Disease).

 

[Glammyre]

Just curious, but why do you want to know about Lyme disease and veterinary medicine? Your questions seem oddly specific and aren't something I'd expect a medical student to ask without cause. I'm all for One Health, but I'm confused about what you want to know and why.

 

[WhtsThFrequency]

No, I am just trying to determine if real world practice relies heavily on microscopic examination of live blood samples & in particular with Borrelia b. (Lyme Disease).

We usually use immunological tests for Lyme disease. Look up the 4dx snap test. Although You can definitely diagnose things like Anaplasma spp, Mycoplasma spp, etc, from a smear. Borrelia too with a nice Giemsa stain. And histologically, if it is a necropsy case and you sample kidney, a silver stain will make the spirochetes light up. But the snap tests are the first line of defense.

Microscopy is obviously essential in veterinary pathology as a specialty. But the average practitioner may do basic cytology or blood smear depending on their expertise and comfort level as well

 

[Minnerbelle]

My microscope is the number 1 used in house diagnostic tool by far, though not for Lyme. We run dozens and dozens of ear and skin cytologies, urine sediments, fecal direct smears, and needle aspirates on any given day.

 

[CalliopeDVM]

How important is a microscope in veterinary medicine for diagnosis of pathogens?

If a dog/cat owner comes in and tells you that their pet had been bitten by ticks & that they are just not acting like themselves, with malaise & perhaps joint pain/stiffness, do you attempt to diagnose by microscope?

Can the Borrelia burgdorferi spirochetes be visualized via microscopy?

A Google search will tell you that Borrelia isn't easily seen with a light microscope. In clinical practice diagnosis (of Lyme) is done with serology, not microscopy.

How important is the microscope for diagnosis of pathogens? Very important, though it doesn't yield a specific species of bacteria or fungus, just the presence of it. Different clinics will use it a lot of a little, depending on their location, as well as the number and skills of the staff. Usually not for blood-borne pathogens (that's usually done with serology) - it's used more for urine and fecal samples, ear swabs, skin sampling, and aspirates.

 

[dyachei]

I use my microscope multiple times a day. skin and ear cytologies, fecal floats and directs, blood smears, urine, masses, skin scrapes, etc etc. I have never used it to diagnose lyme disease but I am a GP.

 

[fastuno]

Anecdotally, I have heard that veterinary medicine microscopy is far more advanced & veterinarians are more likely to incorporate it as a diagnostic tool (as compared to the human counterpart).


I was hoping that diagnosing Lyme via microscopy would be standard practice behind closed doors in the veterinary world.


I have a vested interest because I have always been fascinated with pathology. My family & I have also been bitten by ticks & some of us have manifestations of Lyme Disease. After I asked around I discovered that there were 20 people just on my block who were bit & some who have Lyme, & the numbers grow outside our block.


I started to develop a few unsuspecting Lyme symptoms & I thought to myself that I should be able to see the spirochete in my blood, if Lyme was what I had. After all we are dealing with a bloodborne disease right? A tick does not hatch with Lyme (or so the experts say), it has to bite a mammal or reptile & it has to pickup Borrelia burgdorferi from the blood meal. Which means that the spirochete should present itself in the peripheral blood in fairly large & scattered quantities, in order to propagate the infection through the vector?


So I broke out my microscope & I started checking my own blood for days & weeks, but I did not find a single spirochete, where I was so sure that I would. It just so happens that I was checking while I was on IV Rocephin for Lyme. I thought to myself "they" must be right & that the bacteria are deeply imbedded in the tissues.


When I stopped the IV Rocephin, week after week I would develop new symptoms & I started to see these spirochetes in my blood at last. Each month I was in worst condition & each month I saw more & more spirochetes in my blood. They always remain hidden & invisible within the Red Blood Cells (RBC's). After a few hours the spirochetes burrow out of the RBC's & they can finally be seen in the blood plasma. In the plasma I have also managed to perform time lapse video of them undergoing pleomorphic transitioning so as to prove they are not artifacts. I have caught them in the act of cysting, burrowing out of RBC's, thin borrelia birthing, formation of "String of Pearls" or blebbing/spore formation, & looping and spiraling into each other to form a double strand. I did all this work in 2011 & then started making some of my video & work available publically.


I also set out to find others who might have seen the same thing, I couldn't be the only one who was witnessing this? After months of searching I finally found some people from all over the world, who were sick as I was & who are finding the same thing in their blood. And finally I found two scientists who went publically with a paper & who went on film stating that Bb can be found in the blood. Where my technique was to allow the blood to sit & conditions to change (PH drop, temp drop, & O2 levels drop) and force the bacteria out of hiding, their technique allowed them to "blow up" the RBC's & make them translucent.


Here is the paper on their work.

http://counsellingme.com/microscopy/MysterudAndLaane.pdf


I was hoping to learn more than I knew about Borrelia & microscopy from a veterinary stand point.

 

[dyachei]

from a veterinary standpoint...we do ELISA tests for lyme in combination with other tick borne diseases as part of a screening panel. Then, if the dog is displaying clinical signs, we treat for lyme disease (and send in a sample for titer confirmation). If the patient is not showing clinical signs and there is no current history of tick exposure, we also ask about vaccine status because it can cause a positive ELISA.

 

[WhtsThFrequency]

I was hoping that diagnosing Lyme via microscopy would be standard practice behind closed doors in the veterinary world.

Light microscopy diagnoses of Lyme disease in peripheral blood is massively inefficient in terms of both time and accuracy, unless 1) the particular patient has high levels of circulating organisms, which is rare anyway - many infections may show little to no circulating presence because they have settled in to the deep tissues 2) you are very lucky and just happen to catch said patient above with circulating organisms, because dissemination via peripheral circulation is a transient event. Can it be done? Yes, with enough luck and the right stain. Would I use it as a primary method? Hell no. That is why other methods such as ELISA (or even better Western Blot) are preferable - there will be far too many false negatives with microscopy for this particular pathogen.

You should also know that chronic Lyme disease is a fairly rare condition that many people with other issues such as chronic fatigue, etc. tend to think they have because of stuff they read on the internet. I highly doubt tons of people on your street actually have true chronic Lyme disease.

 

[fastuno]

Thanks for all the responses & I appreciate all the info. It really gives me a sense of how Lyme is dealt with in the veterinary arena & goes against my initial speculations.

Are the ELISA & Western Blot notoriously know to produce a lot of false negatives in the animal world, as they are in human testing? I would imagine so, or perhaps since animals can't speak of symptoms that more owners are accepting of a negative diagnosis?

Elisa & WB tests for only ONE single variation of Borrelia, where in fact there are 100 Borrelia species in the USA & over 300 in outside the US. Not all of them are reported to cause diseases, but neither was Borrelia miyamoto, which was just newly discovered, & which has made its way into the USA. So many go around with a false negative test & yet who have Borreliosis.

I find it easy to find borrelia in my blood & in some of those with Lyme. Some barely have any symptoms & yet there is a plentiful supply in their blood, which then forces one to surmise that Lyme is not Lyme until additional factors are also present. Perhaps cytokine & inflammatory responses, an auto immune system turning against itself, or perhaps co-infections or opportunistic tertiary infections. Perhaps even contributions from environmental toxic factors as well? It reminds me of the whole notion of when HIV+ becomes AIDS & there are immune factors that place Lyme from a passive disease to a more active & debilitating disease.

There has been work now from Advanced Labs & a prominent MD turned researcher that puts live blood culture of borrelia in the high yield zone from a simple blood sample. In fact a standardized culture test has been in the works. Here is a video of a colleague of mine who is culturing blood directly from blood samples using a very simple method.




Some of the newest work with bb involves highly specific DNA Hybridization & probing techniques, & is particular and highly reliable for a Borreliosis diagnosis.

 

[CalliopeDVM]

It seems to me you're mistaking infection with disease.

 

[fastuno]

Nope, I am not mistaking it, but simply stating an observation. I don't know the answers to this & neither do other better trained scientists & researchers yet.

As always, they need more funds to further do more studies.

Keep in mind that sometimes a simple observation (such as mine) can send another researcher into a very valuable tangent.

 

[dyachei]

This is starting to sound a lot like those naturopaths that find "chronic Lyme" in everyone's blood. Not saying it is, but it reminds me of that

 

[DVMDream]

 

[fastuno]

I am not, but I could understand how it could seem that way.
Again thanks for all the insight!

 

[WhtsThFrequency]

OP is determined to be purposefully obtuse. Of course ELISA and WB have failure rates. But so does microscopy for the reasons I stated above, unless we are talking about an acutely infected person with unusually high numbers in the bloodstream, or a badly relapsing person with the same.

 

[WhtsThFrequency]

Two other issues of note. While there are indeed non-pathogenic species of Borrelia, implying that we should desire a test that could detect positivity for all species and/or strains is silly. Because you can bet that what follows will be a lot of unnecessary treatment at the insistence of uneducated patients and paranoid doctors. OP has been reading too much Horowitz. Diagnosing "chronic Lyme" is still extremely controversial.

Secondly, one of the biggest differences between human and veterinary medicine is cost. You can do all the fancy diagnostics on the humans and insurance will foot the bill. We use ELISA snap tests because they are cheap and have a decent measure of sensitivity when the results are put in context with exposure and clinical signs. We can't expect our clients to shell out hundreds or thousands for culture, Western, PCR, FISH, etc nor can we rely on microscopy alone for the reasons I stated above.

 

[fastuno]

"Of course ELISA and WB have failure rates."
Thank you for at least admitting that, as many are still steadfast in their belief that the test is definitive (& yes, human docs especially).


"But so does microscopy for the reasons I stated above, unless we are talking about an acutely infected person with unusually high numbers in the bloodstream, or a badly relapsing person with the same."
Here is a study that takes blood from Lyme victims early on & off season. They are able to use the blood for RNA PCR & confirm a borrelia infection. This means it has to be in the blood, albeit perhaps not in large quantities & not well suited for microscopy. I agree that early infections are most likely not worth the time. It is the long term & chronic ones, that have been sitting around with a negative diagnosis, that can possibly holster the rewards of microscopy.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4139787/


"Two other issues of note. While there are indeed non-pathogenic species of Borrelia, implying that we should desire a test that could detect positivity for all species and/or strains is silly. Because you can bet that what follows will be a lot of unnecessary treatment at the insistence of uneducated patients and paranoid doctors."
I agree. BUT, lets take this example.
-Patient comes in with odd ball symptoms that overlap Lyme & no joint pain.
-Patient (animal or man) receives negative or inconclusive blood tests & you deem safer not to treat.
-Patient comes back in months with degrading health & more overlapping Lyme symptoms & still no apparent joint pain & a 2nd Lyme test mimics the 1st.


As a veterinarian you are not sure of a Lyme diagnosis, do you still prescribe antibiotics at the risk of causing resistance & damage?
Reports are actually coming in from all over the world that the spirochetes can be seen via microscopy if you vacuum seal the wet drop preparation & wait a few hours, then they start exiting the RBC's. There is now even a readily available & low cost preparation to balloon the RBC's in order to see the spirochetes directly within the RBC's (I provided you with the paper & recipe). Does your curiosity not inspire you enough to even check via microscopy? It seems like it is easier to just send them out with their Doxycycline or tell them your sorry since the test came out as it did?


Take a gander at this scientist who was stricken with mysterious symptoms & a negative Lyme tests that did not afford him abx. As is typical with many Lyme sufferers they were bounced around from doctor to doctor with no answers. He had nowhere to turn, but toward his microscope & look at what he is finding. Two pages worth of symptoms & a plethora of bacteria in his blood that he can culture. This is not rocket science, & he can easily see that his Lyme symptoms is caused by massive amounts of spirochete bacteria present in his RBC's. So rather than being in the dark without microscopy, it afforded him the ability to take matters into his own hands & take the next step.



"OP has been reading too much Horowitz. Diagnosing "chronic Lyme" is still extremely controversial."
Many paradigm shifts are controversial in the beginning & many don't really care to look past what they have been told is the correct protocol to follow. It is the easier road to follow.



"Secondly, one of the biggest differences between human and veterinary medicine is cost. You can do all the fancy diagnostics on the humans and insurance will foot the bill. We use ELISA snap tests because they are cheap and have a decent measure of sensitivity when the results are put in context with exposure and clinical signs. We can't expect our clients to shell out hundreds or thousands for culture, Western, PCR, FISH, etc nor can we rely on microscopy alone for the reasons I stated above."
I can agree with expensive tests on animals, but how expensive is it for you to look under a microscope the next time this type of case (as I listed above) comes along? Would you have the curiosity to check?

 

[LetItSnow]

Many paradigm shifts are controversial in the beginning & many don't really care to look past what they have been told is the correct protocol to follow. It is the easier road to follow.

B.S.

The truth is that most paradigm shifts are nonsense and only the occasional one turns out to be valuable. So for anyone actually practicing medicine, it's incredibly inefficient, and poor medicine, to invest heavily (in terms of time, energy, etc.) in "out there," widely unaccepted view points.

 

[Minnerbelle]

I can agree with expensive tests on animals, but how expensive is it for you to look under a microscope the next time this type of case (as I listed above) comes along? Would you have the curiosity to check?

I think you need to realize that Lyme manifests differently in dogs than it does in people. "This type of case" like yours isn't clinically relevant in dogs. So no, I do not have the curiosity to check. And please don't try to tell me that it is clinically relevant, because it really really isn't. In the grand scheme of things, if that were the case, I'd have to check blood via microscopy on every single patient that walks through the door. Like half of my patients as it is are Lyme or Anaplasma positive on an ELISA at any given time and asymptomatic as it is.

Sealing a slide of blood and waiting hours and looking might sound easy and cheap to you, but in our setting I can assure you that is not the case. I would charge at least $80 for that kind of service, and it just wouldn't happen, plus it sounds like a huge pain in the ass. I literally have no time for that. If I have a pet with clinical symptoms of the general tick-borne illness ( fever, lameness, malaise), the workup without microscopy is already at minimum $200 if I strongly suspect tick-borne and I'm not testing for other things. It's actually one of the cheapest workups I can do for a sick pet as it is (Mini CBC/Chem profile plus Lyme/Anaplasma/ehrlichia). You have no idea how hard it is to convince owners to already let me do these things when "they know it's Lyme because all their other dogs have had it." Even if neg, if I still strongly strongly suspect it, the patient will go on a therapeutic abx trial. Tick-borne dogs respond rapidly, like usually within the first 48 hrs of treatment if not the first 24. I don't think my choice to do a therapeutic trial will change regardless of whether I see spirochetes in blood under microscopy or not. And I would under no circumstances use that instead of ELISA because I'm looking for other tick-borne illnesses as well.

 

[Minnerbelle]

I think you should maybe take up your idea to the medical forums and have it entertained there... I know you came here because you'd figured it would be more welcomed since vets use microscopes more often, but clearly you're not getting much traction here. Like I said, "chronic Lyme" in people is not relevant to dogs. Borrelia is actually a very fascinating organism and there is a lot we don't know about it. While doing a lab animal externship at MIT as a student, I heard a leading borrelia researcher talk about what we know and don't know about the org. But a lot of the mysteries are much more relevant in people clinically than they are in dogs.

 

[CalliopeDVM]

"

Reports are actually coming in from all over the world that the spirochetes can be seen via microscopy if you vacuum seal the wet drop preparation & wait a few hours, then they start exiting the RBC's. There is now even a readily available & low cost preparation to balloon the RBC's in order to see the spirochetes directly within the RBC's (I provided you with the paper & recipe).

So what? Again, you are mistaking infection with disease. If I have an ill patient without signs of Lyme disease but with Borrelia spirochetes, it does not follow that the those spirochetes are causing the illness, nor does it follow that the patient has Lyme disease.


As has been pointed out, Lyme disease in dogs is different Lyme disease in people. And our ability in time and money is different than it is in human medicine. If only I could do whatever I am curious about! Oh, what a fantasical joy that would be! The truth is that vets don't get to do whatever they like just because we are curious. We can only do what our clients allow us to do and what we have time to do - it's human medicine that has the time, money, and staff to do whatever they want. We're not House MD, with unlimited money from insurance companies and unlimited lackeys to spend their time satisfying my curiosity. We work without definitive diagnoses all the time in veterinary medicine because of the limited patient cooperation, time, and money (mostly money) that we have. I'd like to see an episode of House where he had just $300 to work with for the work up and treatment of a problem.

 

[WhtsThFrequency]

Does your curiosity not inspire you enough to even check via microscopy? It seems like it is easier to just send them out with their Doxycycline or tell them your sorry since the test came out as it did?....

I can agree with expensive tests on animals, but how expensive is it for you to look under a microscope the next time this type of case (as I listed above) comes along? Would you have the curiosity to check?

I don't understand why you continue to fail to see my point. I would not prescribe doxycycline in the absence of clinical signs. My own dog tested positive for Lyme via ELISA but she has no signs and I'm not throwing doxy at her.

It has nothing to do with curiosity. It has to do with the fact that microscopy is not, and should not be, a screening test with any sort of predictive value in Lyme disease in the absence of confirmation by other methods. If you happen to catch it in the blood, great. But if you don't that doesn't mean anything. It is not consistent enough.

In addition since you brought up PCR - of course Lyme does transiently appear in the bloodstream. No one here is arguing against that although you seem to think we are. But do you have any idea of the tiny number of organisms required to be able to detect with PCR? A number that can very, very easily be missed by a single visual inspection of a blood sample. It sounds like you are trying to advocate blood smears as a primary diagnostic tool for Lyme diagnosis.

 

[CalliopeDVM]

I don't understand why you continue to fail to see my point. I would not prescribe doxycycline in the absence of clinical signs. My own dog tested positive for Lyme via ELISA but she has no signs and I'm not throwing doxy at her.

Exactly - we treat the patient, not the lab results. If the patient doesn't have clinical signs, you don't treat it!

 

[fastuno]

Thank you so much for all the wonderful comments & I understand the vets point of view a whole lot better now.

The only issue I have is with the comment that I am mistaking infection with disease, which I am not really. I see the spirochetes in some of my family's blood, although they are asymptomatic overall. It has been known for quite some time & new evidence comes trickling in that many humans have Lyme, get treated & still continue to harbor the spirochetes. Some have no or very little symptoms. One mother in particular "cured" herself of Lyme (in remission) & had no symptoms. She ended up passing the disease on to her newborn some years later after her treatment.

Thanks again, this has been very valuable to me. I will come back to elaborate some more & perhaps ask a few more questions.

 

[CalliopeDVM]

...I see the spirochetes in some of my family's blood, although they are asymptomatic overall. It has been known for quite some time & new evidence comes trickling in that many humans have Lyme, get treated & still continue to harbor the spirochetes. Some have no or very little symptoms. One mother in particular "cured" herself of Lyme (in remission) & had no symptoms. She ended up passing the disease on to her newborn some years later after her treatment.

You still haven't shown me that they have Lyme disease, only that they have spirochetes.

 

[WhtsThFrequency]

You still haven't shown me that they have Lyme disease, only that they have spirochetes.

And not even Borrelia. Could be syphilis, for example. If they even ARE spirochetes he is seeing, given that he has minimal medical training thus far.

 

[Minnerbelle]

And not even Borrelia. Could be syphilis, for example. If they even ARE spirochetes he is seeing, given that he has minimal medical training thus far.

maybe it's stain precipitate.

 

[cowgirla]

maybe it's stain precipitate.

or maybe it's not on the slide at all... Eye floaters, anyone?

 

[d2305]

One has to have a dark field microscope to see TP.