Faecal examination technique

[tonymaccs]

I have a sick RT female I bought as part of a pair about 2.5 months ago. She is around 5 inches and they were very robust and preparing to spawn when I bought them. Stupidly I skipped QT as I was limited for tank space at the time and put them in a 75 gal with four others. Within a week all were ill with flukes followed by other infections and after several weeks of care all recovered fine except this one. Her only initial symptom was long white trailing faeces identical to photos I've seen on this site which seemed to appear when I treated the tank with Prazi, so I assumed it may have been tapeworm. I noticed she was off her food around this time. These are her only visible symptoms with dark colouration and gradual emaciation.
I've had her alone in a 12 gal hospital tank with a large sponge filter for over a month and she hasn't eaten in that time despite being tempted by a range of frozen and live food. I treated her with Metro (as bath) for two weeks with no visible improvement. I followed this with anti-biotics for two weeks which seemed to improve her general activity level and appearance but still not taking any food.
Water parameters are- stored rainwater, very soft, pH 6.4. Ammonium registers occasionally but I do 70% WC daily. Nitrites not detected, nitrates very low up to 5.
I have a semi-decent microscope but I haven't done a faecal slide as she doesn't pass very frequently and by the time I see it it has been lying around the tank for a couple of hours. Will there be anything lurking behind after this time? Any advice on the best way to do this please?

[tonymaccs]

I forgot to mention- her eyes appear to be very large and bulging but she has lost a lot of mass around the forehead etc which might be accentuating this.

[Graham]

Hi How did you know they had flukes?

Good thing is that you have a scope...so catch the fish and take a slide or the cover slip and draw it along the fish directly behind the gill and down along the side of the fish, You want to pick up some mucus all along the way ....but don't mark the fish. Then add a drop of water to the mucus and add the cover slip to it....set it aside

Then take a second slide or cover slip and draw it down through the chin area, directly below the mouth. Then draw it across the anus.Again add a drop on water and the cover slip.

You've now got scrapes from all the high potential areas...start at 40X then to 100X and then to 400X...flukes are very visiable at 40X. A healthy fish wil have nothing cruising around within the slide

The same thing can be done with a poop sample, but is has to be taken as fresh as possible. There are tons of ''critters'' that will attach it as soon as it comes out of the fish.

This will give you a bit of an idea as to what might be going on

[tonymaccs]

Thanks Graham, I've done the slides and can't find anything of interest externally.

[pcsb23]

Tony,

The fish does not look well. A lot of what you describe is symptomatic of an internal parasite infection such as nematodes or an internal bacterial infection. Prazi won't deal with them, nor does metro in most cases. As its not eating you will need to use something like levimasol or flubenol to treat for nematodes. However the feces suggests an internal bacterial infection. What anti biotic did you use?

[tonymaccs]

Thanks Paul,
I've been heading down the path you suggest since the metro treatment didn't show any improvement. I didn't treat her specifically with Prazi, she just happened to be in the comunity tank I was treating for external parasite symptoms.
The antibiotics I used were erythromycin and cephalexin. Her appearance and activity has improved since, but by no means cured as she is not eating and still passes the white faeces.
I will try a faecal slide, and have been trying to get my hands on Levamisole and flubedazole. The latter is not generally available in Australia but I think I can get levamisole from farm suppliers. Years ago I used to use fenbendazole for internal worms with fish but I'll have to read up on it's relationship with flubendazole etc.
Tony

[tonymaccs]

I managed to get a reasonably fresh poop sample from the sick fish and make a couple of slides. I'm a rank amateur at this by the way so I cant identify what I'm looking at, but I managed to get some reasonably clear pics- see how they turn out after cropping and resizing etc.
The poop is teaming with bugs of two kinds easily seen at 50X mag. The most populous bug is pear shaped and uniformly round. It moves around freely but not so quick. The closest picture that I can find on the Net is that resembles it is Tetrahymena but this is only going on general shape etc.
The second less popular bug but still in numbers easy to find is about 3 times bigger. It is oval shaped with a sharp pointed tail spike and a flattened body convex/concave when viewed from the side (copepod??). It moves much more quickly and erratically than the other so it was difficult to get it clear in photo, but pic 2 shows a couple of them fairly clearly.
Did a couple of comparative samples of poop from my "healthy" discus. The most common bug in the "sick" sample is present but in much lower numbers- I had to go searching for them. Couldn't find any of the second bug at all.
Any thoughts thanks?

[tonymaccs]

A couple more pics at 200X mag showing the bug with the sharp tail.

[Graham]

Hopefuly this willl work. At 50x I don't think that you're seeing anything pathogentic. This tetrahymena

http://www.fototime.com/74215ADE17B3C96/conv.wmv

[brewmaster15]

Hi Tony,
I agree with paul's thoughts here as well.. That kind of fluffy white feces in discus is usually attributed to bacterial infection, at least as a secondary cause. Worms may be underlaying.. Look for barrel shaped eggs for capillaria worms in the feces.

You can use the fenbendazole for worms, but it absolutely can't be added to the water... since the fish isn't eating... gavage (tubing it) is the best way with that med... levamisole can be added to the water.. another possible choice is piperazine.

I'd try to clear up the possible infection first..
Can you get any of these..?.

oxytetracycline
minocycline( found in maracyn2 by mardel)
Kanamycin sulfate
gentamycin

-al

[tonymaccs]

Graham,
That's what I am seeing so it's pretty conclusively Tetrahymena. I gather it's non-pathogenic. Curious though why it's teeming in the "sick" samples taken on different days, but scarce in the "healthy" samples taken under similar circumstances for comparison.
Any ideas what the other bug with the spiked tail is?

[Graham]

Tony at 50X I don't think that what you're seeing but I'm not there seeing it;) I also don't think it's your main problem

This is the post by Johnson concerning the video...

================================================== ==

As replied to by Dr. Johnson:
------------------------------------
That's GREAT footage of a not-too-common pathogen of Koi. It's Tetrahymena. Very cool, because people can mistake it for swarmers of Ich.
Ich swarmers are the organisms which come from the Ich packets on the pond bottom, in the usual presentation, there would be swarmers AND adult encysted Ich in the scrape. In the video
there are not.

Secondly, these organisms demonstrate VERY CLEARLY the highly refractile macronucleus positioned in their wide, tail end.
(They move forward by their "pointy" end.

So, this is superb video, rare-excellent video of Tetrahymena.

I looked on NI and did not see the post - I wanted to ask for
permission to use it on Koivet.com and Fishdoc.

Erik


Tetrahymena is functionally on par, in several ways, with
Trichodina -

Likes warmer water.
Likes water with higher organic loads.
Hates salt usually.
REALLY, RE-e-e-e-Ally irritating to the fish skin - causing
it's damage more by the fish scratching itself on rocks - than
by actually 'eating' the surface.

So, it will make skins pink and fish to scratch and flash.
Other than that, it's not a killer, in my experience.

Of course, forward to Mr. Law and I'd be pleased for my
response to go the NI way. It's not a bad bunch at all. No
worries.

Best regards

E

[tonymaccs]

A bit (lot) more research might be necessary to reduce my ignorance to this issue.
Graham,
I wonder what magnification the video is at? Gives me the idea to use the video function on my camera.

Al,
Haven't yet seen any thread-like worms or eggs/cysts but will keep taking samples.
I've acquired some Levamisole, Piperazine, oxytetracycline and tetracycline. The others you mention are probably prescription only in Oz so will take me a day or so to get if I want to avoid standard Vet consultation fees.

Can the wormers be used in conjuction with the anti-biotics?

thanks, Tony

[Graham]

Tony, , the odds are it was at 400x. For it to be compared to ich swarmers it's small. Here's ich at 40x

http://www.fototime.com/5B3C65AD156B73C/standard.jpg

[brewmaster15]

Good info Graham!:)

Tony,

although I doubt that a deity would have either male or female genitals. Theres really no need.... Worms can cause problems like these, but worms are rarely in high enough numbers to rush into a deworming..

My thoughts on the issue... treat with oxytetracycline for 10 days and see how the fish responds... I have no idea if this is bacterial..but have treated similar successfully. Hopefully the fish starts to eat and then you can add the oxy to the food.

Thats just what I would do, hopefully its of use to you.

take care,
al

[paletka]

Al,
Wher I can get levamisole?.

Chris

P.s.
I need liquid form of levamisole and Metronizadole, do you know if I can get this in USA

Thank you

[tonymaccs]

Graham/Al,
Thanks for the continued help/interest with this. After considering your advice, I did quite a few more microscopic slides last night including a much fresher poop sample and it appears the bugs I was seeing at 50X are probably attaching and multiplying after the faeces exits from the fish. Up to 500X is not yielding anything of interest apart from the odd solitary organism.
It's a pity that a lot of the information on the internet is incomplete or not comprehensive, eg The informative IFAS pictorial guide states the size of the organism in micrometres but would be so much easier if the magnification of the photos and sketches was mentioned.

I will start a course of oxytetracycline which raises the question of doseage for bath. 750-3780mg/10gal is mentioned in one IFAS paper. The dosage needs to increase with water hardness so I presume the lower end for discus water. Does this sound right?
I am very interested in administering by injection as I can access the equipment. Problem again is dose rate. One source mentions 5mg/kg body weight IM, another 10mg/kg as a single dose.
This one mentions 200mg/kg in 50:50 propylene glycol/saline administered IP.
http://www.springerlink.com/content/t28xu71xw766624j/
I wont do this unless I can find something more definative.

Tony

[brewmaster15]

Hi Tony,
You can't inject all forms of antibiotics...The oxytetracycline should be absorbed by the gills okay.. its best in feed though.. .. I have used 500 mg/10 gal... dose daily after 75% wc .. Lights out as this med is oxidized by light...

If you have very hard water you may want to up the dose a little.

One other note..the fish can be skittish... keep the tank well covered.... hth,
al

Chris,
pm sent.

[tonymaccs]

Went the in vitro route, probably a little high on the dose rate at 2000mg/50litres. Hasn't bothered the fish so far, but it oxidised pretty bad by the third day so I will reduce the light levels.