Feline Haemobartonella/Mycoplasma

Re: Feline Haemobartonella/Mycoplasma

Charlotte Marshall — Thu, 05 Sep 2013 22:23:18 GMT

Sorry, forgot you have already done FeLV/FIV test but I would still suspect something else going on as well.

Re: Feline Haemobartonella/Mycoplasma

Charlotte Marshall — Thu, 05 Sep 2013 22:21:47 GMT

There are 3 (I believe) different types of haemoplasma and one of them is more likely to be causing clinical illness than the others. I forget which is which, I always have to look it up as I still think of it as Haemobartonella as well. It is also often found in association with other illnesses so if you are not getting improvement with doxycycline I would suspect a concurrent illness such as FIV.

Re: Feline Haemobartonella/Mycoplasma

John Flynn — Wed, 04 Sep 2013 23:08:08 GMT

[quote user="robloxley"]Are the lab doing a manual or automated retic count?[/quote]

I never thought to ask - had assumed manual prior to you asking.

[quote user="robloxley"]IIRC NMB once made up can degrade relatively quickly.[/quote]

I could be going wrong there then as I don't use it often and purchase a 100ml bottle of NMB 1% in isotonic phosphate buffer ready-to-use retic stain probably only every 2 to 3 years... (£12.90 from http://www.clin-tech.co.uk/catalogue/Clin-TechUk2013.pdf at the same time as getting a new batch of "Speedy-Diff")

[quote user="robloxley"]Obviously don't count heinz body cells or other artefacts (e.g. stain precipitates) as retics! [/quote]

I don't tend to filter prior to use (adds more fiddle and time...), but think I can spot the difference between the stain precipitates and actual retics.

I'd rather rely on a lab, but 20mins to look at blood smears/retic counts on a Saturday evening can be well spent if actually gives some answers (if really lucky I might save 20mins on a discussion with the owner about various differentials and tests for those and what I might find if I was to do these tests etc... sometimes it's just 20mins wasted though for no gain!) that lead to treatment/non-treatment decisions sooner than waiting 3 days for a lab result to return. When we hit the Christmas post, I usually regret not being more familiar with basic lab work...

Re: Feline Haemobartonella/Mycoplasma

Miriam Lodewyks — Wed, 04 Sep 2013 21:11:15 GMT

Thanks guys for your super enthusiastic replies! However, I have neither methylene blue, nor "new" methylene blue, and least of all 20-30mins to prep the slide and examine under a microscope! Assisting the nurses with the venipuncture and filling in the relevant lab-form is as much time as I've got to spare. So I'm afraid I rely on Idexx' pathologist to provide me with relevant information. Still, your posts make for fascinating reading, so keep em coming! PS: have resorted to prednisolone, and Kitty is still alive... For now.

Re: Feline Haemobartonella/Mycoplasma

Rob Loxley — Wed, 04 Sep 2013 00:24:30 GMT

[quote user="John Flynn"]Alternatively there might be quite a bit of inter-observer variability with these manual counts? Or the quality control in the lab I use mightn't be that amazing?[/quote]

Are the lab doing a manual or automated retic count?
IIRC NMB once made up can degrade relatively quickly.
If your sample is anaemic, you'll need proportionally less dye for the same staining
Obviously don't count heinz body cells or other artefacts (e.g. stain precipitates) as retics!

Re: Feline Haemobartonella/Mycoplasma

John Flynn — Tue, 03 Sep 2013 23:35:28 GMT

[quote user="Michael Woodhouse"]If there is significant blue - like in the pics. Not if just an odd spec.[/quote]

Same as myself (other than I mix and incubate my blood and NMB drops in a urine sediment tube for 15-20mins, usually while I clean the microscope lens and examine the fresh blood smear for morphology, before using to make a standard-type blood smear, rather than mixing straight on to slide)

That said, I do wonder about how well I do this as on occasions as when I've sent a blood away and got a retic count, but did one myself first for more immediate info, the counts are sometimes pretty far apart - enough to draw different conclusions quite regularly.

I suspect this just reflects that I don't do these often and counting 1000rbcs (a process I find rather laborious) is neither what I nor my microscope are built for? Alternatively there might be quite a bit of inter-observer variability with these manual counts? Or the quality control in the lab I use mightn't be that amazing?

Re: Feline Haemobartonella/Mycoplasma

Michael Woodhouse — Tue, 03 Sep 2013 23:19:33 GMT

[quote user="John Flynn"]Do you count all blue granuled rbcs in cats as retics for instance? I know some vets that do.[/quote]

If there is significant blue - like in the pics. Not if just an odd spec.

Re: Feline Haemobartonella/Mycoplasma

John Flynn — Tue, 03 Sep 2013 23:03:41 GMT

[quote user="Michael Woodhouse"]It is new methylene blue we have. I thought old methylene blue was the McFaydean's anthrax stain. Doesn't it simply oxidise or something with time.[/quote]

I was just trying to save someone taking up your tip from ordering the wrong stain.

"New methylene blue" has a different chemical structure from "methylene blue" (an extra methyl group on each of the outer benzene rings according to wikipedia!). They are effectively different chemicals, with different staining indications, but confusingly similar names.

In years gone by, I was disappointed that my polychrome methylene blue (yet another variant... McFadyean's stain: an oxidised product of "methylene blue" as I understand it) didn't really give me reticulocyte staining like I expected. That said, I never found anthrax with it either

[quote user="Michael Woodhouse"] I wasn't aware I had to go into intricate detail of the procedure as I thought we all did this?[/quote]

But we might all do it differently!

Do you count all blue granuled rbcs in cats as retics for instance? I know some vets that do.

Re: Feline Haemobartonella/Mycoplasma

Michael Woodhouse — Tue, 03 Sep 2013 22:16:30 GMT

It is new methylene blue we have. I thought old methylene blue was the McFaydean's anthrax stain. Doesn't it simply oxidise or something with time.

You definitely get blue granules in reticulocytes. I wasn't aware I had to go into intricate detail of the procedure as I thought we all did this?

Re: Feline Haemobartonella/Mycoplasma

John Flynn — Tue, 03 Sep 2013 19:01:37 GMT

[quote user="Michael Woodhouse"]

Staining to look for retics is easy. Drop of EDTA blood, mixed with drop of methylene blue. Smear on a slide. The blue RBCs are erythrocytes.

Screamingly obvious if there.

[/quote]

I think you need new methylene blue (or an alternative supravital stain) to stain up reticulocytes.

I expect this is a typo (as is "blue RBCs are erythrocytes"?)

Just trying to save anyone reading money and frustration on a bottle of "plain" methylene blue

Re: Feline Haemobartonella/Mycoplasma

Richard Carter — Tue, 03 Sep 2013 18:29:14 GMT

Isn't methylene blue for staining up heinz bodies?

Also H.felis or M.felis can be incidental finding if rbc non-regenerative anaemia as like most parasites, keeping under control depends on an intact immune system. If is a case of haemobartonella, > 80% rbc have bodies attached. Also make smear from fresh ear tip prick, not edta or heparin as can wash parasites off with anti coag. Stain deposits refract light when focussing up and down, H.felis remains constant blue and should be variations in numbers, some cells with multiple inclusions.

Re: Feline Haemobartonella/Mycoplasma

Michael Woodhouse — Tue, 03 Sep 2013 16:44:00 GMT

Staining to look for retics is easy. Drop of EDTA blood, mixed with drop of methylene blue. Smear on a slide. The blue RBCs are erythrocytes.

Screamingly obvious if there.

Re: Feline Haemobartonella/Mycoplasma

Richard Carter — Tue, 03 Sep 2013 09:10:19 GMT

Vit B inj? If kitty circling looking for the exit would second preds as well as continuing doxy or marboflox if no other money left.

Re: Feline Haemobartonella/Mycoplasma

Andrew Kent — Mon, 02 Sep 2013 21:19:34 GMT

It would be worth another external haematology now to assess for regeneration as it dictates where to go from here. In the absence of any other tests it is difficult to see a good outcome. If they will really do nothing else then could try prednisolone for a pure red cell aplasia but would be doubtful it will help!

Re: Feline Haemobartonella/Mycoplasma

Miriam Lodewyks — Mon, 02 Sep 2013 21:03:56 GMT

PCV today 16%. Clinically unchanged. FeLV/FIV negative. Unlikely clients will pay for any more tests.

Re: Feline Haemobartonella/Mycoplasma

Andrew Kent — Mon, 02 Sep 2013 19:50:08 GMT

I wouldn't count visible inclusions as particularly sensitive as they can be seen with other things (eg artefact, Heinz bodies etc). Would funds stretch to a pcr? This would need to be run on a sample taken before tx and is pretty sensitive.

the absence of response to doxy makes haemobart unlikely so would be thinking of other things. Have you repeated the haem now? Is it still non-regenerative?

If so then you have waited long enough for it to be pre regenerative so would look at the bone marrow and consider felv, neoplasia etc. are any other cell lines reduced?

Andy