Histology of Apisotos

[Noddy65]

Hi all...
My lovely wife has been processing one of my apistos for me and sending me pics. I was thinking I might include the histology slide pics as I get them. This may take a little while as they slides take a while to process and strangely this isn't a high priority for my wife.

1. Gill biopsy. nothing unusual here. The dark staining spiky things are melanophores. The dendritic processes can be very large and almost neuronal like. If you get fresh ones they can have fine cytoplasmic granules that are motile – easy to confuse with pigmented fungal spores. They have phagocytic activity so are part of the normal defence mechanism. Large numbers are an indication of chronic inflammation.
http://i15.photobucket.com/albums/a378/Noddy65/gills1.jpg

[fishgeek]

what power maginification is that?

i am assuming it is the tip of a single gill lamellae?

nice shots it would be nice to accumulate a few of these in one place ..

andrew

[Robdog]

There were a lot of big words there Noddy, so how about a bit of laymans stuff. What are the little black dots that look like freckles or peppering?

And get your wife on the program! Tell her that there are strangers all around the globe that are waiting on these slides. :roll: Sheesh Some people :roll: :wink:

Great photo too. Not really sure what it is but great photo. Was that taken with a disposable camera? :lol:

[Th0mas]

Rob,

These are the kind of word used by doc - Mike (Noddy) is a vet, and my guess is the missus also in similar field of work.

Anyway, great pics - problem is mortal like myself won't understand the meaning of them. Probably better to have a drawn up diagram indicating there the section is from and uses simple word for those who don't have a medical dictionary handy.

My first impression of the photo without reading the text - "What on earth is that has to do with apisto??" :oops:

Cheers,
Thomas.

[Noddy65]

Sorry guys...
Not too sure on the magnification...Ill have to ask the wife.

Ok...its a sample of a gill. Under the operculum (the gill on the outside) is a collection of very fine true gills. What we call the gill isn't really the gill, its the operculum. There are heaps of these very small and fine gills under the operculum, they are responsible for extracting oxygen from the water and exchanging it with CO2 (and I assume a few other waste products). The main thing you can see is that dark thing running down the left of the slide, that is one edge of the gill. The spiky things coming off it are normal gill structure. The dark line ending in a curve on the right side of the slide is the 'inner' part of the gill. The little dark things are the melanophores. I didn't know what they were so had to ask a friend.
Melanophores are used to 'gobble' up infections, they are a normal part of the fishes defence system. If in large numbers, and I think there a large numbers here, it indicates a chronic infection/inflammation.
The clear bubble like things on the right? Hmmm...not sue about those, Im assuming theyre just normal side effects to death ie. tissue leaking fluid.

I'm a small animal vet (dogs, cats, birds etc) and not a pathologist so this is all unfamiliar to me. I just believe what my wife tells me.

She has cut the fish up to do the rest of the body, so hopefully there will be some more nice pics in the next weeks or two.

Mike

[Robdog]

That's as good an explanation as I could get Mike. Thanks.

I'm guessing that due to the size of these little fellas a biopsy such as this would be a fairly tricky procedure and require a very steady hand?

[Noddy65]

Yep...anaesthesia would be preferred, magnification, sharp instruments and a very steady hand.

Mike

[Robdog]

So that fish is still alive?

[Noddy65]

Nope...that fish is deceased..
Wife is sitting next to me...magnification is 200x.
Mike

[Ben]

Mike, great photo mate and please keep them comming!
Very interesting to see this kind of fish photography!

Cheers
Ben

[fishgeek]

mike i would assume that the photo is of a single secondary lamellae?

to me the gill is like combs lying flat on top of each other (these are known as the gill arch's and there can be upto 4)
the teeth of these combs are the secondary lamellae
then i think the hair like projections are gill filaments

other than that i cant make much of it either
andrew

[KillieOrCory]

Wow! Great pic and info!

Can I ask for a favour? Please :wink: If the head is pretty much intact would you ask your wife if she can take photos of the otoliths of your apisto. She would be very busy and have tons to do, so only if she has the time.

They should look something like this:

http://i63.photobucket.com/albums/h139/KillieOrCory/guesswhatthisis.jpg
This set I got out of Aphyosemion australe (a killifish).

If possible seeing the otolith crystals would be very interesting.

[Robdog]

Nope...that fish is deceased..
Mike

Well if the fish is already dead, and that is what I thought might of been necessary to get a photo like that, why is an anaesthesia preferred?
Respect for the dead :lol:

[Noddy65]

HEHEHEHE...no..gills biopsys can eb atken with the fish alive..anaesthesia is required.

[Robdog]

Gotcha!

[Grae]

Thanks Mike
that really is amazing stuff
Thanks to Zoe to.
Now all we need is some EM shots

Graeme

[Noddy65]

More photos soon guys...the wife was very excited about the slides...she cant believe the fish lived as long as it did, apparently is is full of what looks like TB granulomas, for those in the know she is doing a ZN stain today.
Hopefully Ill post some pics tonight.

Mike

[leamos]

Keep 'em comin', reminds me of uni, i 've spent many an hour looking at fish slides, that is the nature of pracs when you're studying aquaculture / marine bio.

[Noddy65]

Sorry everyone...it will be another few days. The ZN stain will be at least a few days and the normal slides? Well she has sent copies off to a friend first before committing to any kind of diagnosis...Im not even allowed to see them...

Mike

[Noddy65]

On another note, I have wriggelrs from the fine fish in my sig...very excited about it too..

Mike

[fishgeek]

if it is tb then get changing all your substrate
and strat isolating any fish with any signs of any illness

good old mycobacterium are a pig to remove
fingers crossed it is cancer or something non communicable anyway

[Noddy65]

Thanks
Ive gone one further and removed all substrate, Im gradually removing it all from my apisto tanks.

All tank furniture is being soaked in hot water.

As an interesting side note, hot water is great for getting rid of snails but Java moss doesn't like it one little bit, even a 10 second dunk cooks the plant :D

Mike

[Th0mas]

I've heard tanks needed to be wiped down with phenol to ensure the mycobacterium is completely gone. Getting hold of these chemical shouldn't be a problem for you though.

Hopefully it's not TB, wish you luck mate.

Thomas.

[Noddy65]

Might have to throw out the sponge filter as well.
A pathologist friend of mine used to keep apistos a few years ago but he eventually gave up. Appears TB is endemic in apistos (at least the ones we get here in Aus). His all eventually succumbed to the disease so he moved on to other things.
I read somewhere that the lower ph can initiate clinical signs of the disease.

Mike

[Th0mas]

Yeah - it seems to be quite common, but how low is low pH?

Thomas.

[fishgeek]

Tb in all aquarium fish has been reported various studies from anywhere from 25% to 60% prevalence

I have not read in any texts the story about water pH having an effect on clinical outcome nor signalment of this paricular infection, prehaps a rumour started by the reading of something about staining techniques(?acid fast stain?!)

Tb is a slow progressive problem and usually takes years to show(lifespan of an apsto?)

best guess therapy that i can find in referenced text would probably be doxycycline and rifampicin as a combination treatment
then again things as varied as cyclosporin!(an immunosuppressive drug?) have been cited in literature

basically the very variable answers for what may work in accepted medical texts makes me believe we just dont know of an effective treatment

not sure prehaps the aqis or whoever is responsible for fisheries in aus may have some more recent data, apparently a lot comes from really old french tb institute work

options for disinfection include
potassium permanganate, hydrogen peroxide and alcohol
peroxide is pretty simple for tanks


andrew

[Noddy65]

I think the low pH thing may have been anecdotal, I read it somewhere on the web (and we all know how reliable that info can be).

Its an interesting disease process from what I can gather.

The clinical course can run from slow weight loss and ill thrift resulting in death right through to sudden death with no/mild previous clinical signs. It makes sense though as it would depend on where and how many granulomas form.Apparently some fish can show cutanous lesions that spontaneously resolve and never reapper.

Although I dont have any evidence I would imagine that husbandry/water quality/temp/breeding status/individual variation would have an impact on the ability of the disease to progress from a latent/carrier stage through to exhibiting clinical signs of the disease.

On an aside...Andrew is that a A. trifasciata in your sig on Apistogramma.com and if so, which type. It looks just like mine and Im trying to ID my fish.

Mike

[fishgeek]

yep trifasciata , and i dont go for all the local bunf.. i generally try for aquarium bred fish , thats my attempt to reduce wild catching

so all the collection locality is just stuff i dont get involved in and subsequently dont know

speaking with a friend whom imports he says he cant see the difference between rio guapore and other trifasciata that are being sent to him, so not sure it is even reliable

andrew

[Noddy65]

Hi all
well the wife has come through alright. The pics are really nice and she's even labelled most of them (she's into training first and second year students at the moment).

Ill try and describe and explain them to the best of my ability, if I'm being too vague then speak up. Remember, I'm not a pathologist.
Andrew may be able to make a contribution here as well.

The diagnosis is about 99% Tb now, she is just waiting for some special stains that show up the TB bug better. Its up to her lab to process the slides and its not a priority.

1. This is a general layout of the slide. Remember that microscope slides are essentially a two dimensional view, there is no/very little depth. This is a slice taken of the fish laying on its slide. If you read the labels you'll get a better idea.
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-label.jpg

2. This is a slide of its head, again just a two dimensional view. The various colours are the stain that shows different tissues. Looking at this view there are already some TB granulomas (TB infection often results in a granulomas which is essentially a walled off area in the body surrounding by fibres with the bacteria living inside. this wall an make treating difficult as antibiotics done penetrate the granulomas walls very well and so cant kill the bacteria). the dark purple bits in the eye are a TB granuloma. Theres a better view to follow.
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-Face-label.jpg

3. Apisoto eye. The dark purple areas are the centre of the granuloma, if you look closely you can see a sort of thin wall around it. The pics arent very detailed as I had to shrink them for the web so don't be too disappointed if you cant see it.
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-Eye-10x.jpg

4. Moving further back into the fish we have its abdomen. The circles area in the middle is a huge area of granulomas, either one large one or numerous small ones in a cluster. this granuloma is taking up an enormous part of the fishes abdomen. About a week before it died it had a 'lump' protruding from its side that shrank over time, I wonder if it could have been this granuloma.
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-Abdomen1-label.jpg

[Noddy65]

5. Out of interest these are the fishes gonads full of eggs :(
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-Gonad-label.jpg

6.Heres is the poor girls kidney with another nice juicy granuloma circled.
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-kidney-label.jpg

7. And some muscle with more granulomas circled.
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-muscle-label.jpg

8. This final one is a close up of a nice granuloma. If you look closely in the darker pink areas there are some patches of paler blue, sort of at he bottom right of the large circular pink thing. this is almost certainly a huge area filled with the Tb bacteria, this should stain really nicely with the special stains that are being done.
http://i15.photobucket.com/albums/a378/Noddy65/Apisto-bacteria.jpg

Well that's all for the moment, Ill include the ZN stain slides when I get them.

To sum up: this female A. panduro almost certainly succumbed to TB. She has large TB granulomas in muscle, kidney, abdoman and even the eye.

[fishgeek]

nice series mike
do you know how they process such large sections?
i though that the microtomes were smaller than that?

andrew

[Noddy65]

Hi Andrew
The specimen was small but the wife has assured me that microtomes can cut quite large section, big enough to fit on a slide anyway.

The fish was large enough to fit on one normal microscope slide.

Mike