Disease diagnosis with the aid of a Microscope

[Alvin Koh]

Dear AroMedics,

I had always wondered how possible it is for a home aquarist to diagnose the various types of diseases via a microscopic view of either the water or a scale scraping.

It would certainly take the guesswork out of identifying the type(s) of diseases that are present in an aquarium. The prohibiting factors would be the microscope - what range of magnification is required to perform a thorough analysis?

It would be ideal if we could ultimately have some form of disease database for non/magnified images for diagnosis purposes.

Regards,
Alvin Koh

[Isna]

It's actually quite feasible Alvin.
Takes a minimum amount of skills n experience to run one of these things. but yeah, cost is a major limiting factor. generally, u'll need a mag of 40 or 60X to b able to diagnose most protozoan bugs. a 100X oil immersion might b useful for bacteria, but not necessary. it's close to a grand to get a decent workable microscope in NZ....would cost about the same i guess elsewhere.

another issue is proper training. u need quite a bit of experience n exposure to b able to recognise n diagnose the bugs. ( there r a couple or so good websites with real photomicrographs of parasites....even video clips...)


But really, at the end of the day, most diseases can b treated satisfactorily without the aid of a microscope. Simply basing your diagnosis on the clinical signs n history will narrow down the list of possibilities greatly.
nowadays, besides skin scrapes/cytology, sick commercial fishes are getting bacterial/fungal cultures, virus isolation, xrays, CT/MRI scans, blood sampling etc..........these r obviously useful diagnostic equipment, but it's really getting a bit complex n specialised. That's why u have fish vets.

so yes, u don't need to have a microscope. it'll b nice if u have one..
:D :D

[Vetduck]

Errr....Isna/Wayne,

Actually the magnification on a microscope is 100x, 400x & 1000x(oil immersion). Although the rotating lens are 10x, 40x & 100x, don't forget the eye piece is another 10x. When you multiply the magnification of the eye piece & the 100x lens, you'll get 1000x magnification.

Alvin, as Isna has mentioned, learning to use a microscope is not difficult. It is being able to identify what you see is what requires training & experience. Experience is what enables one to identify the difference between an artifact & a actual pathology.

Fish medicine is quite a specialised field, requiring proper training. What you see in a fish scrape is very different from that of a dog skin scrape, I can assure you. :D

[Isna]

Hi vetduck,
u r right. the actual magnifications have to include the eyepiece. funny thing is, most pathologists, cytologists, clinicians i know speak of the rotating lens mag only.

anyway, have u had a look at Nauzer's aro's slides?

[^GeNoME^]

where to get those stains to identify those bacteria?

[Nauzer]

Originally posted by Alvin Koh

Dear AroMedics,
It would be ideal if we could ultimately have some form of disease database for non/magnified images for diagnosis purposes.

Regards,
Alvin Koh

Hi alvin,
You mean an image database similar to this:
http://puregold.aquaria.net/disease/bugspics.htm#Chilondonella

though its not an aro-related site, but it does show the magnified parasites that allows some form of comparison and understanding. You are right, if we could begin compiling a database of such parasites by image.....that would be the next big leap for arofanatics.

p.s. the Nikon Coolpix 990, 995 can be attached to a microscope for taking such shots.

[Vetduck]

Originally posted by Isna
Hi vetduck,
u r right. the actual magnifications have to include the eyepiece. funny thing is, most pathologists, cytologists, clinicians i know speak of the rotating lens mag only.

anyway, have u had a look at Nauzer's aro's slides?

Hi Nauzer & Isna,

Had a look at the impression & squashed smears of the nodules.
There were lots of inflammatory cells. Large numbers of macrophages & some lymphocytes. There also appears to be some cocci bacteria. Some epithelia cells & melanin pigments. Large no. of macrophages & melanin pigments would indicate some form of inflammatory reaction going on. Could be of bacterial origin.

I spoke to my friend who is a fishvet & he reckons it could be some form of epitheliocystis or localised bacteria infection. He said that lymphocystis doesn't occur in aros. He feels that localised topical meds, such as topical acriflavin, would solve the problem.

Nauzer, I'll call you over the weekend. We'll try to medicate your baby. :)

[Nauzer]

Hi Vetduck,

Even though i have the slightest clue about the definitions that you have just mentioned, it is comforting to know that it is not the commonly proclaimed anchor worms, fish louse, etc.
I'd honestly rather bacteria infection than parasites!

In all my time of rearing aros, this is truly the highest technical/ medical stage it has ever gotten to, and truly, i appreciate all help you have rendered.

To me, this is a kind of break through news! At least, in terms of arresting problems, this is the most accurate form of logical progression in the manner of treatment.

I will be back from an overseas trip earliest next Tuesday afternoon. Will give you a call then.

Cheers man!

[Isna]

sounds good vetduck!
did u use a dif-quick stain?
perhaps a fresh wet-smear would have been extra useful too just to rule out protozoans etc( can see them moving...), and some authors recommend a routine gill biopsy with every skin scrape.

well, sounds like another nondiagnostic sample to me...really frustrating....i've taken a few nodular smears, hoping to see spectacular pathology......but often times u don't see much.
at least we've ruled out some stuff, but u always wonder about sampling errors etc.

cocci bacteria r exceedingly rare in fishes...most superficial bacteria r long rods---columnaris like bugs. they r often secondary to something else. were these cocci intracellular? could it b nuclear debri from ruptured cells?

i would go for a deeper scraping, and try to debride as much tissue as u can, it'll help with the healing process if it was a localised bacterial granuloma...secondary to trauma? foreign body??

have fun guys!

[Vetduck]

Hi Isna, I just stained it with diff-quik. I'm won't be surprise that there are lots of artefacts on the smear. A canine skin scrape is usually more fulfilling than a fish scrape.

Unfortunately the scrape wasn't very diagnostic. At least it gives us some idea of the nature of the lesion. We'll try to treat it symptomatically & check response to treatment.

I've scraped it quite deep, almost down to the ray. Hopefully it is adequate to make a difference.

Nauzer, sorry for the jargons. What it all means is that we haven't been able to arrive at a diagnosis. All we could suspect is that it is a localised inflammation, perhaps of bacterial origin. What we can do is to try treating & see if it gets better. :)

[Nauzer]

hi vetduck and Isna,

ok.....i see! I guess we would have to eliminate step by step. So what would the treatment require? Acriflavin and a hospital tank?

Is acriflavin a strong potent dose of medication that affects the aros behaviour, appetite,etc.?

[Vetduck]

Originally posted by Nauzer
hi vetduck and Isna,

ok.....i see! I guess we would have to eliminate step by step. So what would the treatment require? Acriflavin and a hospital tank?

Is acriflavin a strong potent dose of medication that affects the aros behaviour, appetite,etc.?

I was thinking more in terms of localised acriflavin application to the lesion. Localised topical application wouldn't affect appetite or filter media.

[Nauzer]

that means another occasion of transmore and treatment? And when we place the aro back into the tank immediately after, doesn't the acriflavin get cleansed away? how does it work?

[Isna]

Hi guys,
how's the aro going Nauzer?

anyway, just wondering about the acriflavine.
i've never heard of topical acriflavine in fishes...but i guess it should work.
povidone iodine is frequently recommended in various texts, but u've got to b careful not to get the healthy skin...use a cotton swab...
Some clinicians prefer panalog to iodine....
personally, i've never tried topicals.....only reserve it for nasty focal infections--possibly HITHD.
have u guys considered a simple salt bath?

[Nauzer]

Hi Isna,
Thanks
Actually i wish i could see my aro! I am currently away on a biz trip. So the closest i could get are verbal descriptions from my wife back home.
I heard some good news from her last night, she mentioned that the infected area seems to have gone down, or hardly an obvious lump. What remains seems to be a only the wound from the scrape. This sounds like good news!
Hopefully, the natural resistance of the fish is combating the bacterial infection on its own. When i get back, i will begin to take some closer shots again and have them posted in this thread.

regards

[Vetduck]

Originally posted by Nauzer
that means another occasion of transmore and treatment? And when we place the aro back into the tank immediately after, doesn't the acriflavin get cleansed away? how does it work?

Hi Nauzer,

The acriflavin would eventually get washed off, but it is not as aqueous as say chlorhexidine, it kinda stains the lesion & stays for a bit longer. Don't worry, don't have to transmore your aro again. Will explain to you when you get back.

[Alvin Koh]

Hi guys/docs, :)

Thanks for the enlightenment, I am sure I'm not the only one with this nagging query with regards to Microscopes and diseases diagnosis.

Nauzer: That database link you provided is indeed the sort of pictures I am referring to. Now if I only I could get my hands on a proper microscope....

Alvin.

[Nauzer]

well as far as i know, Nikon manufactures really heavy duty powerful microscopes as well.......:)

So attach a Coolpix 9 series and off you go.

[Hobbit6003]

Hi Vetduck and nauzer,

haha, histopathology was one of the worst subject I've taken at med school.:D

Vetduck, did you perform an excisional biopsy on the nodule? If so, wonder if they did recurr?

Also, was the fish vet Frederick Chua?

I'll be interested to tag along the next time you're going to do the topical application of acriflavin on the aro, if both of you don't mind.

Cheers,

Kenny

[Vetduck]

Originally posted by Hobbit6003
Hi Vetduck and nauzer,

haha, histopathology was one of the worst subject I've taken at med school.:D

Vetduck, did you perform an excisional biopsy on the nodule? If so, wonder if they did recurr?

Also, was the fish vet Frederick Chua?

I'll be interested to tag along the next time you're going to do the topical application of acriflavin on the aro, if both of you don't mind.

Cheers,

Kenny

Hi Kenny,

Yes the fish vet is Fred Chua. You know him?
Well I just scraped the whole nodule off. Sometimes it solves the problem completely. I haven't seen the aro sinced, if it has healed then probably don't have to do anything.

[Hobbit6003]

Hi Simon/Vetduck,

Yeah, I do know Fred Chua, though not too well. We corresponded briefly while I was doing the 'Ask Naki' section in Straits Times, I've quit since. We also talked briefly when I asked him for advise when I had camallanus worm infestation in my guppy tanks. His brother is also a dental surgeon. I had met his partner Fred Chong, quite a chatty guy!:)

I think you've got good chance of a full recovery when a total resection of the nodule is done. From the description of the histo section, looks like a localised infection of some sort to me. The macrophages, epithelial cells and melanocytes all point to a chronic granulomatous growth.

I guess it is hard to pinpoint the specific bacteria, if it is indeed one, as chances of contamination is high.:)

Anyway, if you are going to visit your patient again, can I tag along as well?:)

Cheers,

Kenny

[Nauzer]

Hi Isna, Vetduck & kenny,

Sure you can join us kenny. After my next few postings and comments from you guys (to treat or not to treat), even if we decide not to treat, we should also meet up to discuss/ view aros and enjoy coffee. :) it would be a pleasure.
Sorry i took ages to reply to this thread, have been away from S'pore. But I am back with some updated images of the nodules 10 days after the scrape process with Vetduck.
As you can see in the diagram below, the nodule seems to have subsided. Previously, it was a formation of small mess of clustered lumps that contributed to a big hump, as vetduck had witnessed. Now its gone down to its original/ initial small nodular swelling.
Please see circled area....

[Nauzer]

another current image, this view gives a better perspective of how raised the nodes are......has indeed gone down a lot......almost flattening out.

[Isna]

hmm, it's definitely still there.
thanks for the update Nauzer. :D
i was thinking, if u guys r planning to knock out the aro to apply topicals etc, might b worth taking another fresh scraping from the lesion, plus some over the rest of the body, and possibly a gill biopsy too. i'm thinking of a wet mount. this way, u can easily see any parasites cause they'll still b moving. we should rule out infectious bugs, cause it'll change our therapeutic approach significantly.
how do u plan to apply the topical without sedation vetduck? might get a bit wet...anyway, have fun guys.:D

[Vetduck]

Hi Nauzer,

Hope you had a good trip. Hmm... it looks like it hasn't resolved completely. Might have to try medicating topically then.

Isna, was planning to restrain it in a bag. Maybe bag it, empty the water gradually, then cut a hole where the lesion is & medicate topically.

Nauzer, will call you over the weekend. Maybe monday afternoon if you're free.