# UPDATE: IT'S FISH TB. ...Help me diagnose this disease that's killing my fish



## FlyingShawn (Mar 4, 2011)

Seems like the pics aren't working. Here they are:


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## wkndracer (Mar 14, 2009)

My best guess would be internal parasite based and Levamisole HCl powder, flubendazole are my 2 choices because fish don't have to eat it.

http://www.plantedtank.net/forums/fish/138137-quarantine.html

good luck OP this stuff when it happens is frustrating


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## FlyingShawn (Mar 4, 2011)

wkndracer said:


> My best guess would be internal parasite based and Levamisole HCl powder, flubendazole are my 2 choices because fish don't have to eat it.
> 
> http://www.plantedtank.net/forums/fish/138137-quarantine.html
> 
> good luck OP this stuff when it happens is frustrating


Thanks wkndracer! I'll order both of those right away.

After the platy in QT died, I was getting scared that it might be Fish TB (based on his symptoms and those other meds not working), so I really hope you're right.


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## beardedcharmer (Sep 24, 2011)

that's a tough one. Could be parasite with a secondary bacterial infection or TB. I like metro combined with prazi for parasites but have also used vermisol and panacur. For bacterial I like kanamyacin and furan-2


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## FlyingShawn (Mar 4, 2011)

beardedcharmer said:


> that's a tough one. Could be parasite with a secondary bacterial infection or TB. I like metro combined with prazi for parasites but have also used vermisol and panacur. For bacterial I like kanamyacin and furan-2


Do you have any recommendations for where to get those meds? I'm probably going to try the ones wkndracer recommended first, but it might be smart to get them and keep them in my medkit so I have them in the future.

@Wkndracer:

Reading through your posts in that quarantine thread, I saw you suggested the possible use of an HP treatment for bacteria/parasites. Do you think that'd be worth trying in the meantime while I wait for the meds you recommended to arrive?


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## Jeffww (Aug 6, 2010)

Is the ONLY symptom bloating? Just bloating seems odd. Really without any obvious symptoms you can't know for sure without some autopsies which require a microscope and some veterinary knowledge. Do you think you could cut one open maybe if one dies? We need to see what's swollen, the swim bladder, intestines, stomach etc. Do they have trouble swimming? When did this start to occur? Add any new fish or plants? 



> The first victims were a platy and some neon tetras


The platy suggests something parasitic. Common livebearers are almost ALWAYS disease ridden animals. 

I want to say parasites but I can't be sure. Try feeding some peas to clear any intestinal blockage and see. Observe their poo. Its coloration and texture (extremely thin, pale etc.) can tell you a lot. 

I can suggest you get some garlic and mash it up a bit to get lots of good garlic juice and soak the medicated foods in the garlic. It should make them take it a little better. Plus garlic is naturally antiparasitic so there's that bonus as well.


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## FlyingShawn (Mar 4, 2011)

Jeffww said:


> Is the ONLY symptom bloating? Just bloating seems odd.


Actually, the bloating is only very slight (no more than you'd see after them having a bigger than normal meal). The _primary_ symptom is the near-anorexic appearance of the body aft of the stomach, where it seems to totally collapse in on itself. Take a look at that first picture again: it's not that the thinness of the aft body just looks that way in proportion to the bloat, it's that the width of the bloat is only _slightly_ more than the normal body shape and the tail has collapsed that much!

Coloration stays near-normal up until death, as does appetite. The infected fish will become somewhat more reclusive and skittish. There are no visible symptoms other than the ones mentioned above that I've been able to identify.

The disease first appeared in a long-established tank that hadn't received any new introductions (plant or fish) for at least a few months. That said, I can't be totally sure that it wasn't introduced by something because of the extremely slow progression of the disease: one fish at a time, dies slowly, then a few weeks later another begins to show the same symptoms.

The first platy infected had been in my tank for 2+ years and the second was actually the ~1.5 year-old offspring of some platies I had at the time, so I think I can safely discount them as the original source.

Thanks for the tip on the garlic!


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## wkndracer (Mar 14, 2009)

FlyingShawn said:


> @Wkndracer:
> Reading through your posts in that quarantine thread, I saw you suggested the possible use of an HP treatment for bacteria/parasites. Do you think that'd be worth trying in the meantime while I wait for the meds you recommended to arrive?


H2O2 works for external issues only to my knowledge from reading threads on TAFF II and the linked study threads included in Garrett's posts on that site. Safe concentrations / ratio data varies not just by fish species but by users reporting to use it so caution going forward with that. After my desperate acts using it I read all that I could find on the topic. (still will use it again too)
Found that Otto's don't tolerate it well either.

Salt baths are also an external option and can help with gill related parasites. 

Kanamyacin is reported (idk) to be about the only thing to help with TB being a strong antibiotic.

Angels Plus sells medicated food that contains it.

also most things are available here (watch price LOL);
http://www.fishyfarmacy.com/products3.html

fyi lots of stuff has a shelf life along with being expensive so research that too stocking a war chest.

good luck OP


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## beardedcharmer (Sep 24, 2011)

aquabid has the best price on kanamyacin (mvp), angelsplus has the best prices on the prazi and metro and jehmco is a good source for the panacur (flub)


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## inkmaker (Jul 11, 2006)

FlyingShawn said:


> Thanks wkndracer! I'll order both of those right away.
> After the platy in QT died, I was getting scared that it might be Fish TB (based on his symptoms and those other meds not working), so I really hope you're right.



Let us know how things go when you get the meds. Pictures if possible.

Charles H


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## inkmaker (Jul 11, 2006)

beardedcharmer said:


> aquabid has the best price on kanamyacin (mvp), angelsplus has the best prices on the prazi and metro and jehmco is a good source for the panacur (flub)


Panacur is Fenbendazole not Flubendazole. Flubendazole is absorbed buy the skin and gills of the fish and does not have to be eaten to be effective.

Charles H


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## beardedcharmer (Sep 24, 2011)

yes my bad its fenb., you can dose fenb in the water column and it is effective against more parasites than flub. you just have to be more careful with the dosing


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## sewingalot (Oct 12, 2008)

*Beware of fenbendazole!*



beardedcharmer said:


> yes my bad its fenb., you can dose fenb in the water column and it is effective against more parasites than flub. you just have to be more careful with the dosing


I strongly, strongly, strongly (I can't emphasize this enough) recommend against *fen*bendazole. I killed at least 43 fish using half the dosage recommended by wkndracer. I had mistaken *flu*bendazole for the same as *fen*bendazole. You can see the damage here (http://www.plantedtank.net/forums/1342425-post1341.html), how it doesn't disolve and also if you continue reading (back up a few pages for the beginning of the deaths), you'll see that it happened within hours and the deaths didn't stop for days. I lost almost everyone of my fish due to using this.

To the OP, that does look at lot like internal parasites, but the sunken stomach makes me worried that it's TB. Not to be gross, but how is their poop? Is it white and stringy (a good sign of pests) or is it normal looking?


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## FlyingShawn (Mar 4, 2011)

I ordered the Leva and Flub from Charlie today. Based on what wkndracer said, I'm not planning on trying the HP while I wait for it to arrive.

If these two don't do the trick, do you have any recommendations for the order to try the others? I don't want to go nuts and order everything listed here (for the shelf life reasons mentioned above), but I need to be thinking of a plan of attack now (both an order of operation and knowing which are safe to combine vs must be dosed individually).

Aside from the Leva and Flub, you guys have suggested Metro, Prazi, Kanamyacin, Panacur, Vermisol, and Furan-2. I'm guessing the Kana should be next (in case it's TB), but I'm wide open to suggestions since I've never used any of them.

Thanks again!


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## wkndracer (Mar 14, 2009)

inkmaker said:


> Panacur is Fenbendazole not Flubendazole. Flubendazole is absorbed by the skin and gills of the fish and does not have to be eaten to be effective.
> 
> Charles H


Very happy to see you post on the forum Doc!

Ladies and Gentlemen Charles H. Harrison, Ph. D.,,,
a man who's saved more fish than my well water has killed.


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## FlyingShawn (Mar 4, 2011)

sewingalot said:


> I strongly, strongly, strongly (I can't emphasize this enough) recommend against *fen*bendazole. I killed at least 43 fish using half the dosage recommended by wkndracer. I had mistaken *flu*bendazole for the same as *fen*bendazole. You can see the damage here (http://www.plantedtank.net/forums/1342425-post1341.html), how it doesn't disolve and also if you continue reading (back up a few pages for the beginning of the deaths), you'll see that it happened within hours and the deaths didn't stop for days. I lost almost everyone of my fish due to using this.
> 
> To the OP, that does look at lot like internal parasites, but the sunken stomach makes me worried that it's TB. Not to be gross, but how is their poop? Is it white and stringy (a good sign of pests) or is it normal looking?


Just to clarify, do you think Fenb is just that dangerous that is should be avoided or do you mean you used Flub dosing guides with Fenb by mistake (so you just have to be careful to keep them straight)?

I haven't had the opportunity to observe their poop due to my work schedule and not knowing what to look for. I'm off tomorrow, so I'll catch him in a breeder box, feed him, and keep a close eye on him. I'll let you know what I find.


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## wkndracer (Mar 14, 2009)

fenbendazole has to be eaten to work and the raw chemical added to tank water while several have said they do it has real risks.
Sewingalot did it by accident others claim it to be practical, cheap, easier to find and safe (imo) not my tank.
Mixed with food products yes (easier to buy it that way though)


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## garfieldnfish (Sep 25, 2010)

It looks like dropsy to me and metro meds may help if your fish will eat it.


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## Jeffww (Aug 6, 2010)

garfieldnfish said:


> It looks like dropsy to me and metro meds may help if your fish will eat it.



Dropsy is marked by raised scales as well and a much faster decline..

Also, what temp do you keep your fish at btw?


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## FlyingShawn (Mar 4, 2011)

Jeffww said:


> Dropsy is marked by raised scales as well and a much faster decline..
> 
> Also, what temp do you keep your fish at btw?


I'm very confident that it isn't Dropsy. The bloating is too minor and it doesn't match the wasting which is the primary symptom.

I keep the tank at 78F.

I just got home (been away a few days for work) and found a couple of interesting things in my war chest (you know, it really helps to have some idea what names to look for):

-Jungle's Anti-Parasite medicated food apparently contains Prazi! I tried putting some in before I left a few days ago and it seemed everyone tasted and rejected it, but I'm hoping that'll change tomorrow since they haven't eaten in a couple of days and I'm going to try Jeffww's trick of soaking it in garlic (I'm also going to crush some of it to make it bite-size for the Cardinal Tetras in the tank).

-I also looked at the ingredients on Tetra's Parasite Guard tablets and discovered they contain "Praziquantel, Diflubenzuron, Metronidazole, and Acriflavine." Unless you guys see a reason not to, I'm thinking I'll go ahead and dose the tank with these while I wait for Charlie's meds to arrive.


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## wkndracer (Mar 14, 2009)

-Jungle's Anti-Bacteria or Parasite medicated foods are really low on my list but hey starve them long enough and fish will eat wet cardboard LOL

I always try not to create a toxic soup with too many meds. at once. Clean water and good O2 is important having it's consideration in treatments too


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## FlyingShawn (Mar 4, 2011)

wkndracer said:


> -Jungle's Anti-Bacteria or Parasite medicated foods are really low on my list but hey starve them long enough and fish will eat wet cardboard LOL


That's exactly what I'm hoping: between the hunger and the garlic, it'll at least get _something_ into their systems.



wkndracer said:


> I always try not to create a toxic soup with too many meds. at once. Clean water and good O2 is important having it's consideration in treatments too


That was exactly my concern when I was asking what to combine vs dose individually. I figure the tablets can't make too much of a soup on their own and Charlie's instructions for the Flub start with "Change out as much water as you can before treatment," so I figure most of what the tablets add will be removed then.

If I begin to get concerned about the O2 levels, I can hook up an pump to the air lines/stones I ran under the substrate when setting up the tank (when it takes this long to set up a tank, you have a lot of time to plan ahead for this sort of contingency!)

BTW, thought you might like to see the tank as a whole. I picked out the Noah's Ark decoration for our first family tank when I was 5 and have displayed it in my main tank ever since (22 years!), so I decided to make it the centerpiece of the new tank's scape and title it "Mount Ararat" (sorry about it looking so bright, I was running out the door when I took the pic and didn't turn the exposure down far enough):


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## wkndracer (Mar 14, 2009)

haha like the story! thanks for sharing that.
The tanks I have that are filtered with a canister I clean the can about a week before I do my worming and the water changes both before and after have the critters belly dragging the gravel. 

hope you get this all resolved so the drama ends.


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## FlyingShawn (Mar 4, 2011)

I guess they're not as hungry as I was hoping: even with the garlic I got little interest in the Jungle food. Oh well, they'll get there eventually.

I put the sick on in a breeder box for a couple hours to observe his poop. It seemed pretty normal to me: 0.75-1.0cm long, greenish-brown coloration.

I've now dosed the tank with the Parasite Guard I mentioned earlier (including letting one of the tablets dissolve in the breeder box to give him a little-bit stronger dose before releasing him again).

Now I guess it's just a wait-and-see until I get Charlie's meds.


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## sewingalot (Oct 12, 2008)

FlyingShawn said:


> Just to clarify, do you think Fenb is just that dangerous that is should be avoided or do you mean you used Flub dosing guides with Fenb by mistake (so you just have to be careful to keep them straight)?
> 
> I haven't had the opportunity to observe their poop due to my work schedule and not knowing what to look for. I'm off tomorrow, so I'll catch him in a breeder box, feed him, and keep a close eye on him. I'll let you know what I find.


To clarify, wkndracer (Mike) gave me the instructions for the flubendazole. I only had enough for half of the dosage recommended, but still used it. However, I made a horrible mistake and it was actually *fen*bendazole that I had on hand. (Messing up two little letters is a bad, bad thing.) Besides the mass posioning, I knew something was wrong just by the undissolved white mess in the tank: http://i252.photobucket.com/albums/hh38/sewingalot/Aquarium/20110329184337.jpg

From what I can gather is many of the fish swallowed the fenbendazole whole. Many of the dead fish had it in their digestive system. I found out in a disgusting manner, the link I gave in a prior post shows the half of a fish that I examined. Plus, you could see the lumps on the whole fish. Between eating it and it being in the water not dissolving, it really did a number on the tank. And after the inital loss of like 35 fish, they kept dying for days. I have used 0.1 grams in a 10 gallon to get rid of hydra in a _shrimp only tank _with success. But, as you can see in the above picture link, it didn't effect the shrimp despite it being everywhere. I can no longer recommend it for anything other than hydra in a shrimp _only_ tank. And even then, I don't recommend it further.

As far as *flubendazole*, the stuff that Mike recommended, I have used it successfully to treat velvet and the fish did wonderfully. I wouldn't hestitate to use it in the future. It was a miracle drug as the stuff I brought for velvet wasn't safe for the rest of the tanks inhabitants including the plants.

I have used Jungle antiparasite medication (the tablet kind that foams) successfully for fish in the past, but it took a couple of treatments. It was really gross to see the worms that came out of the danios that were sick. I've heard it doesn't work for every parasite, though.

(Edit - I just saw your last post that you already checked the fish. You are a ninja! ) Definitely check the fish's poop if you can. If you ever see white stringy stuff, they are definitely sick with parasites. Now the tricky part is seeing good ol' poop and knowing this doesn't mean they aren't sick. 

One more thing. Be careful not to mix medications and to give the fish time to recover before adding more medications and be sure to change the water between treatments. For example, you could treat the fish and they would be free of the parasites, but still have a sunken stomach and a little bloating This is also due to not getting the food they needed. Think of getting a bad cold. You are over the cough and snotty nose, but you are still needing more time to recover from the fatigue. I'd work with only one medication type at a time, wait at least a week or so after treatment. Feeding them, watching them closely and then go from there. 

I like your tank and it's story by the way. Good luck on the fish. Please let us know what happens.


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## driftwoodhunter (Jul 1, 2011)

Sorry for a derail, but I'm finding this interesting - I used fenbendazole on my 55 tank that I had to break down. Not knowing any better, I used it (based on recommendations) for a possible planaria outbreak. Now I think they might not have been planaria. Anyhow, it was days later when my fish started dropping like flies from columnaris. I wonder if the fenbendazole could have weakened the fish's immune system? (They ate a great deal of it, and I treated with it for days)
Again, apologies for the derailment...


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## BostonPete73 (Dec 3, 2011)

Hi all,

I am new to the forums, but I have been searching obsessively to find the answer to this same question. My fish have this SAME disease I have been wracking my brain to figure out what it is. Just like the OP said, my fish have slight bloating in the front, but then their rear is really droopy and wastes way. Color and appetite stay normal until the end, but in the very last stages (right before death) the wasting becomes so pronounced that the fish have difficulty swimming. Some fish will last for up to a month while others will be dead within a week, and it seems like whenever one fish is moved to quarantine another one has become sick.

Some info about my tank, it is 16 gallons up and running for six months now, heavily planted, with three guppies, four platys, some tiny fry, and an unknown number of cherry shrimp and MTS (recently added four assassin snails and the problem began about two months ago). Temperature stays between 73 and 74, Ph is a little bit above 7.2.

I am convinced that it must be something internal, as if it were environmental, it would affect more than one fish at once (imo). I am really worried now that it is fish TB, so I have been trying to remove fish that are showing symptoms as soon as possible, but as my tank is cooler than most tropical tanks, is this even possible? I've heard that _Mycobacterium marinum's _ideal temperature is closer to 30 degrees celsius. I've observed poop and it is definitely NOT white and string, it looks the same as it always has. Anyway, I will try some of the medications that previous posters have suggested, but THIS IS SO FRUSTRATING!


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## FlyingShawn (Mar 4, 2011)

BostonPete73 said:


> Hi all,
> 
> I am new to the forums, but I have been searching obsessively to find the answer to this same question. My fish have this SAME disease I have been wracking my brain to figure out what it is.


Sorry to hear that Pete.

At this point, my plan of attack is to continue with the Parasite Guard tablets, then try the Leva and Flub from Charlie, and then move on to the Kana (since it's rumored to help with TB). I'll keep you posted on what happens, but results are going to be slow in coming.

Since we're both paranoid that it's TB and I've already got the Leva and Flub on the way, would it be worthwhile for you to start with the Kana instead, as a sort of multi-vectored attack?

Let us know what happens.


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## FlyingShawn (Mar 4, 2011)

I've now completed two doses of the Jungle tablets with no noticeable improvements (which is what I was expecting, since I'd tried them with the platy that died before). 

I've given up on the medicated food also. Even after the garlic soak and nearly a week of not being offered anything else, they barely touched the stuff (it was also something that I'd tried with the platy). At this point, I think it'd be better to go back to the regular food (Tetra's tropical crisps are the main staple, with some freeze-dried bloodworms tossed in occasionally) than keep starving them on the medicated food.

Charlie's meds arrived on Monday, so tomorrow morning I'll do the 95% WC and administer the first dose of Flub. I plan on doing two doses of Flub per the instructions on his page, then have sewingalot's recommended break for a few days before dosing the Leva.


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## wkndracer (Mar 14, 2009)

Do you have a plan on mixing the flu?
The powder is a PITA to get into solution without premixing.


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## FlyingShawn (Mar 4, 2011)

wkndracer said:


> Do you have a plan on mixing the flu?
> The powder is a PITA to get into solution without premixing.


I guess I hadn't put too much thought into it. Do you think it would work to put it in a large Ragu bottle about 4/5 full of water, seal, and shake vigorously? I could potentially mix half of it at a time if it's as bad as you're saying.


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## FlyingShawn (Mar 4, 2011)

You were right: that stuff is a PAIN to get into solution. Even with my trick of premixing 1/4 Tsp at a time in a Ragu bottle, I still spent the next day and a half stirring the top of the water to try to mix it in.

I've just finished the first 3-day Flub treatment and started the second stage (5-7 day) treatment. Unfortunately, the Odessa seems to be continuing to go downhill rather than improving. He still has a healthy appetite (although seems to spit food out more than the others), but he seems to still be losing weight.

Also, there's a small area on his side where he seems to be missing scales and the white-spot on his forehead is now more of a grey color and has raised edges (maybe the outer white layer of his skin at that spot fell off?). In some ways, it's reminiscent of hole-in-the-head disease, but I'm wondering if that would just be a secondary infection due to whatever this primary issue is.


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## Jeffww (Aug 6, 2010)

Try adding some sodium hydroxide to get it dissolved....

Mix 40g naoh with 1L of water then add it drop by drop to the dry powder...then dilute the whole thing into some distilled water.

Otherwise you could make a large batch of very dilute solution and perform a water change with the treated water (this method is by far safer and easier lol).


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## wkndracer (Mar 14, 2009)

Dosing into my tanks a HOB method works if you have one. Place the meds behind a floss panel and the water flow will break it into solution in about 2 days. Getting the flu into solution and then into the water column is the biggest challenge of applying the treatment.

I settled on a combination of heat and alcohol after contact with Dr. Harrison.
Dosed dry the first time it took days for it to stop floating in the tank on the waters surface.

Alcohol (vodka) is mixed into the measured dose in a glass container. Only enough to make a paste or slurry out of the flu powder around 1ml more or less, just enough to wet it. Heating a cup of water to boiling in the microwave I wait until the boil settles then add the water in on top of the slurry. Some foaming occurs during the mixing but the majority of the material mixes very well doing this. The liquid is milky in the mixing cup and the tank becomes milky / cloudy but the dilution created by the tanks total volume seems to allow the chemicals to be absorbed into solution and remain suspended by all the evidence following a treatment. My tanks are clear the following morning. Fish have recovered every time I've used the treatment with the exception of two long infested adults. Those fish died with a bloody discharge seeping from the vent. Assuming to much internal damage due to long term exposure to the parasites is my belief. Those losses happened during initial treatment of my tanks when first learning about Flubendazole.

Many find the information being collected in my quarantine thread to be helpful. (this is posted there)


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## FlyingShawn (Mar 4, 2011)

Thanks for the advice guys! As I said, I've already put the 2nd dose into the main tank (and will just have to keep stirring over the next couple days), but I'll definitely keep these ideas in mind when using it in the future. That HOB method sounds especially easy for dosing my new arrivals in QT.


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## FlyingShawn (Mar 4, 2011)

DISASTER!!

The second dose of Flub is still going (nearly over) in the main tank (and everything seems fine, which makes the following that much more baffling):

I decided to use the flub on the new arrivals in my QT tank, as recommended in wkndracer's QT thread above. The QT is a 10gal mostly-bare tank (has some colored-glass pebbles covering the bottom, a couple of fake plants, and some frogbit floating about) with a heater and Tetra internal filter (cycled media from another tank). In QT, I had five ~2" Rainbows (3 Bosemani, 2 Turquoise) that I've had for over a week and were doing excellently by every measure I have (great color, appetite, activity level, normal poop, etc). 50% WC's were done every 2-3 days, with ammonia staying consistently low (not quite zero on my continuous-indication meter, but well below the next increment of 0.05ppm). I was not testing nitrite or nitrate because of the frequency of the WC's and how well the fish were doing.

Last night I did a 90% WC (per the flub instructions) and dosed the flub into the tank (may have been _slightly_ overdosed, but by a really tiny amount like a 9.5-10gal dose in ~9gal of water volume). Took a while to get the flub to dissolve, but it did.

Here's where it gets weird:

On my main tank, after the flub dissolves, the water is clear. After the flub dissolved in the QT, the water was cloudy.

By morning, the water was still cloudy, but the rainbows seemed ok (however, they didn't have much of an appetite when I fed them). Charlie's instructions for the flub say to have "live wiggling or jumping food" for the fish as they recover, so I suspected that flub might cause a loss in appetite and began to wonder if I was doing something wrong with dosing the _main_ tank, not that something was wrong in the QT.

When I got home from work this evening, I noticed the Rainbows were swimming at the top of the still-cloudy water and got concerned. I was trying to figure out if I should abort the treatment and do a WC or if I had time to post a thread and ask about it.

45 minutes later, I saw two rainbows laying upside-down on the substrate and not moving. I immediately jumped to action by netting all 5 fish and putting them into a guppy tank nearby, but it was too late for the two on the bottom. BTW, they look completely fine, even had good coloration when they died. The other three are stressed, but seem to be bouncing back.

A closer look at the water shows that it's not just cloudy, but there are long white stringy filaments floating about and clinging to the outlet of the filter (you can see it hanging off the fake plant in the picture). It's almost like a fungus you'd see grow on food left at the bottom of the tank, but there's way too much of it and it's too long to have grown overnight. None of this stuff was in the tank when I did the WC last night!

The presence of the white-stuff and how dense the cloudiness is leads me to doubt that it's a bacterial bloom from the large WC (and the slight bloom I had when I started the QT was far less than this). UPDATE: I just did a test on the water and couldn't even detect a trace of nitrite.

What could this be?! If it's related to the flub, I'm going to be far more hesitant to use it in the future.


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## wkndracer (Mar 14, 2009)

Wow and sorry to read this reported. Plz send Charles an email and ask him what he thinks but it looks like the dose was higher simply based on the amount of clouding captured in the picture. Every time I've dosed flu the tank has clouded but cleared overnight and the effect on water clarity was more a haze and not the heavy clouding I see here.
Did you have an air stone in the tank?


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## FlyingShawn (Mar 4, 2011)

wkndracer said:


> Wow and sorry to read this reported. Plz send Charles an email and ask him what he thinks but it looks like the dose was higher simply based on the amount of clouding captured in the picture. Every time I've dosed flu the tank has clouded but cleared overnight and the effect on water clarity was more a haze and not the heavy clouding I see here.
> Did you have an air stone in the tank?


That's part of why I'm so baffled.

The dose being too high would explain the cloudiness (but considering there's zero haze in my main tank, this isn't the magnitude of effect one would see from a maybe-10% OD) or a lack of O2 suffocating the fish would explain the deaths (since there was no air stone, just the HOB), but neither would fit the explosive-growth of the white stringy material.

There's enough of it covering the bottom of the tank that I suspect it's the main cause of the cloudiness, but I can't figure out what it could be that it would take over the tank that quickly.


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## wkndracer (Mar 14, 2009)

Five 2" Rainbows is not an insane number of fish but I don't have enough information on whats happened here to do anything but guess.
(and I've thought about it since reading the post)

What is your water source? Is it treated tap, was it aged? Did the large WC done pre-treat go normally?
The stringy material possibly released slim coat off the fish? Stressed fish can produce a ton of slime and any ammonia reading is always a bad thing.
7days plus with low ammonia readings mentioned did it spike at the end after adding the flu and the WC? Parameters can flip fast on small tanks and fungus growth can be explosive covering huge areas overnight so I wouldn't rule that out completely either.

again sorry you lost fish


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## inkmaker (Jul 11, 2006)

FlyingShawn said:


> That's part of why I'm so baffled.
> 
> The dose being too high would explain the cloudiness (but considering there's zero haze in my main tank, this isn't the magnitude of effect one would see from a maybe-10% OD) or a lack of O2 suffocating the fish would explain the deaths (since there was no air stone, just the HOB), but neither would fit the explosive-growth of the white stringy material.
> 
> There's enough of it covering the bottom of the tank that I suspect it's the main cause of the cloudiness, but I can't figure out what it could be that it would take over the tank that quickly.


WOW! I'm surprised as well and everything doesn't fit together. 
The drug does scavenge Oxygen.
How about aeration and tank temperature? Do you keep the water surface moving?

Charles H


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## FlyingShawn (Mar 4, 2011)

After considerable thought and some feedback from Charles, my working theory is that suffocation was the cause of death of the fish. I couldn't find it in any of the articles or instructions that have been linked to, but apparently flub is an "oxygen scavenger."

I think what happened is that the filter on the QT (a Tetra internal model, I think it's the 30i) provides adequate circulation to aerate the water for the fish, but not enough for the fish+flub (especially at higher dosage levels, which leads me to the next point).

I also found out from Charles that the flub instructions linked to on wkndracer's QT thread are outdated, because since publishing them he's switched to a different supplier and is now selling flub in a 10% concentration, rather than 5% as before (he's currently working on an updated set of instructions to give out). Because of that, I ended up dosing my tanks at approximately 2x the normal amount (he mentioned that it's difficult to OD flub because it drops out of solution quickly when the temperature drops).

My main tank is still fine, with clear water and healthy occupants, but the oxygenation rate is substantially higher since it's only half full and my canister outflow is spraying against the back wall of the tank about 6 inches above the water level. The QT however, since it probably only had adequate aeration to begin with, wasn't able to cope with the oxygen scavenging level of a 2x flub dose (BTW, both tanks are kept at 78F).

So, the moral of the story: when dosing flub, ALWAYS use an airstone (even if aeration is normally adequate). Charles, please include this point in your new instruction set.

I still can't explain the white stuff, but wonder if it might be related to a mass death of the nitrifying bacteria due to the O2 levels.

I'll keep you posted on the progress of the main tank.


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## wkndracer (Mar 14, 2009)

What was linked in my QT thread were articles published online by Charles as you say in your post. After reading your last post and what is the probable cause of your losses I reviewed all my posts in that thread with intent on removing ANYTHING that could cause harm or misunderstanding.

In post #18 I did detail what I settled on doing to aid in mixing flubendazole. Reviewing my thread I found he (Charles Harrison, Ph.D.) has already updated the linked information dated 12/16/2011. Included in it also is additional mixing instructions that all (imo) will find helpful.

I am NOT a vet or have I ever attended any medical training classes other than basic first aid and CPR training. Included in my QT thread it's stated that what I shared was based on my personal experiences and input mainly from breeder related websites. That information was spread out within the posts made. I have added an edit to the opening post of the thread in hope of avoiding misunderstandings.

I am truly sorry that trying to share information that I learned through hard lessons has led another person to deal with killing they're pets.


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## FlyingShawn (Mar 4, 2011)

wkndracer said:


> What was linked in my QT thread were articles published online by Charles as you say in your post. After reading your last post and what is the probable cause of your losses I reviewed all my posts in that thread with intent on removing ANYTHING that could cause harm or misunderstanding.
> 
> In post #18 I did detail what I settled on doing to aid in mixing flubendazole. Reviewing my thread I found he (Charles Harrison, Ph.D.) has already updated the linked information dated 12/16/2011. Included in it also is additional mixing instructions that all (imo) will find helpful.
> 
> ...


Please don't take any offense wkndracer: I never meant to imply that you had any role/blame for what happened in _any_ way, shape, or form. I only meant to convey that you linked to the instructions Charles had posted (which were accurate at the time) and that in the time since you made the post, he's changed suppliers and the instructions had changed (which you would have no way of knowing).

Nor to I blame Charles: he'd included a quick summary of the instructions in an email, but I didn't notice the _tiny_ detail of a change in recommended dosage (from 1/4 TSP per 5 gal to 1/4 TSP per 20 gal, which I'll admit I don't quite understand since he only went from 5% to 10%, unless he's also decided a weaker dose is equally effective). He had no way of knowing that I'd use the instructions linked to from your thread, so I wouldn't have expected him to say not to use them.

The responsibility in both cases is mine alone.

I wanted others to know that those instructions were out of date so that they wouldn't follow them like I did and so that they'd know that Charles was working on updating them. I also wanted others to know that flub is an O2 scavenger, since neither set of instructions mentioned it and I thought it an important detail. From now on, I'm always going to use an airstone when medicating (and even when not medicating on the QT) to ensure that aeration is never again a variable at play.


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## wkndracer (Mar 14, 2009)

no offense at all my friend. I just went back and made certain that I had the information posted in that QT thread as clear as possible and added a disclaimer.
There are countless variables in this hobby and nobody ever knows all of it, living is learning.


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## FlyingShawn (Mar 4, 2011)

Time for an update:

I've finished the second dose of flub (full seven days) with no improvement. The original sick Odessa ('1' from here on out) continues to eat well, but has also continued to lose weight and his colors have paled a little more.

After I finished with the flub, I did a three day does of Levamisole, which finished today. '1' seems to have slightly less stomach bloat than before, but is still _very_ thin.

I've also noticed what might be an early symptom in another of the Odessa males ('2'). He hasn't lost any weight yet, but he does seem to have a _slight_ swelling or bloat, but only on his right side. The swelling is farther forward on the body than '1''s stomach bloat, so I'm not sure what's going on. You can see where the bump is located in the first two pictures and the lack of it on his left side in the third.

I've attached the best pictures I could get of each (I had an EXTREMELY difficult time getting even these shots: they were friendly and swam up to me _until_ they saw the camera (flash off) and immediately ran away. It's almost as though they knew what I was trying to do and didn't want to help). '1' was less skittish than '2' around the camera, but it was still hard to get a shot showing how much weight he's lost (the best indication you'll get is looking above his stomach in the last picture and seeing how narrow the body is right under the dorsal fin).

Since the Leva dose ended, I've done another ~90% WC and now have the tank 100% again. Between visiting family for Christmas and other commitments, I won't have enough time at home to dose any more medication for the next two weeks or so, so they're going to have some time to recover from the meds. I'll continue to feed generously to make sure '1' is able to get enough food in spite of the extra chasing-around the others have done since he started losing weight. If the Leva did the trick, he should start showing a visible improvement soon. 

In the meantime, I'm going to be ordering some Kana to try in a couple weeks.


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## FlyingShawn (Mar 4, 2011)

It's time for a long-overdue update (sorry, been really busy with work and some other things):

When I got back from my Christmas trip, I found that the fish labeled as '2' in the pictures above had developed the worst case of Dropsy I'd ever seen. He was so big it seemed like many of his scales were nearly perpendicular to his body! Since I didn't have the Kana yet, I decided to treat the tank with Maracyn II and feeding them with broccoli in hopes of dealing with this new problem.

After a week of dosing the tank with the Maracyn II with no noticeable improvement, I moved both of the sick fish to a 10gal quarantine tank with a divider to keep them from harassing each other.

I've now been dosing that QT with Kana for two weeks using the dosing instructions for internal infections found here (targeting 100ppm active):
http://www.kordon.com/aquavet/kanamycin.htm
I know it only says to treat for 3-5 days, but another site (http://www.fishyfarmacy.com/products2.html) says to use Kana for "up to 30 days" for TB.

My dosage has been 2.25 TSP Kana in a 10gal QT, with 50% water changes and ~1.25 TSP more Kana every three days (slightly ODing on the Kana there, but that first link implies that a little too much shouldn't be a problem and I doubt it'll do any worse to the fish than the disease).

So far, there's been no significant improvement in either fish. Fish '1' still has a good appetite and his weight loss seems to have leveled off, but he's not gaining any either. '2''s dropsy varies in severity from day to day, but averages a little less than where you see him in the pictures below.

At this point, I'm not sure the Kana is doing these guys any good, but until I reach the 30 day mark or decide what to do next, I'll keep using it. I did pick up a bottle of PraziPro at the sorta-local mega-LFS (TFP in Lancaster), so I'm debating trying that next (either by stopping the Kana or by trying both simultaneously).

As always, I'm wide open to any suggestions or insights you may have!

Here are the updated pics. Sorry about the net for the last couple, '2' felt more like hiding than getting his picture taken...


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## FlyingShawn (Mar 4, 2011)

Bump/update:

The fish are still alive and their condition is unchanged from my last post.

At this point, I've pretty much given up on any chance of them recovering, but have continued to treat/care for them in the hopes of learning something that'll help me save the rest of the tank. Currently, none of the fish in the main tank are showing symptoms, but this disease moves so slowly I'm not drawing any conclusions from that.

I decided to look into the possibility of performing a necropsy on them and finally finding out the real problem. After making a thread about it in the Pennsylvania sub-forum and some time spent Googling, I may have found a solution:
Necropsy near Harrisburg?

The quick version is that the vet would perform a necropsy on up to three fish for $100 and look for internal parasites and mycobacterium. If he saw signs that made him suspect TB, it'd be an additional $80 to have samples sent off to a lab and cultures performed.

Steep, I know, but I can't keep up this guessing game of buying expensive meds and hoping they work. If I'm finally able to know what I'm actually up against, that would give me the ability to either treat the real problem or make some very hard choices about the future of this tank. However, if I end up nuking the tank anyway, I'd have $180 less to put into rebuilding it.

What do you guys think? Given all the time and money you've put into your tanks, is that knowledge (good or bad) worth $100-180? At this point, I'm leaning towards having the necropsy done.


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## FlyingShawn (Mar 4, 2011)

I decided to follow through and have the necropsy done on those two Odessa Barbs. The short of it is that the vet is highly confident they were infected with _Mycobacterium_ (Fish TB). In my explanations below, I'll stick to the prior naming convention of '1' for the fish with all the weight loss and '2' for the one who had Dropsy symptoms.

After discussing the results, we decided to go ahead and send off the samples to a histopathology lab to confirm that it was _Mycobacterium_ and not something else (according to the vet, there are very few other things that could cause these results under the microscope). Doing the histopathology will have the added benefit of seeing if there were any other bacterial or parasitical infections going on that we didn't see (including identifying the one nematode parasite he found).

At this point, I should put the disclaimer that all the pictures you're about to see were taken by and are the property of Dr. Brian Palmeiro and are posted with his permission and encouragement. The explanations, however, are based off my recollections of what he told me and any errors can thus be attributed to me (I welcome correction of any such errors, but if you're harsh in doing so, I'm liable to respond with an inverse-McCoy: "Dammit Jim, I'm a pilot, not a veterinarian!").

I'm including more slides than are directly relevant to this diagnosis, mainly because I thought they were really interesting and I thought you guys might appreciate them (but at the same time I apologize for the extra-length post as a result).

How about we start with the only bright spot of the day? Here is a section of 2's gills:

Have you ever heard how the ammonia in an uncycled tank can burn the gills of the occupants? The result of that is a thick coating that develops around the tissue and reduces the gill's ability to exchange gases, which is what makes this image notable: there's no such coating. Dr. Palmeiro said that these are the healthiest gills he's observed in a very long time and that it's evidence that my water quality is very high.

Now, onto the bad news, here's a section of 2's kidney:

The large dark masses are the granulomas that are a characteristic fingerprint of TB. As you can tell, there are a significant number of them. In case you're like me and first noticed the branched dark-spots with hard edges instead of the more rounded granulomas, those are apparently pigment centers and are normal.

Here's another section of 2's kidneys:

Maybe it would be more accurate to say that this is a picture of where 2's kidneys _should_ be. The granulomas have taken over to such an extent that 2 actually had almost no functioning kidney tissue left. This, not a secondary infection, is the reason for the dropsy: because the kidney wasn't functioning to expel fluid, it was building up within his body and causing the bloat I'd been observing (you know, I didn't know that fish pee. It makes sense, I just never stopped to think about it before!).

1's kidneys weren't as extensively damaged, so he was still able to expel that fluid and avoid the bloating. Before today, if you'd asked me which of the two was in worse shape, I'd have guessed 1 over 2 nine times out of ten. Shows what I know...

In case you were wondering, here's a not-so-bad section of kidney to compare to (this is from fish '1'):

Note: the less-brown background color of this one is evidence of what I was saying earlier about a fish needing to be _very_ freshly dead to give good data: it had been barely 1.5 hours since they were euthanized and there was already a notable deterioration of the tissue! The transparent tube- or worm-like structures are "mineralized tubules," an aging change in fish.

Moving on, here's a section of 2's spleen:

There are two things visible here: the same sort of granulomas caused by _Mycobacterium_ that we saw on the kidneys and what's called "inflammatory centers." I can't positively remember which is which, but I believe the darker ones with the harder edges are the inflammatory centers). As you can see, the damage was pretty extensive.

For contrast, here is a spleen section from '1':

He still has the inflammatory centers and granulomas, but not nearly to the extent that '2' did.

Here's a section of 1's GI tract, specifically the intestines:

The circular things you see are actually food particles that were still in his system when he was euthanized, showing that he continued to eat up until the point we put him down. As a somewhat related side note: contrary to some of the articles I'd found in my research that suggested loss of appetite as a symptom of TB, Dr. Palmeiro says the the massive weight loss and continued appetite till death or very near death is actually quite common.

Zooming in on a different section of 1's intestines, we found a single nematode parasite:

A single parasite does not a major health problem make, so the going theory is that this single parasite did little to contribute to 1's health problems. What I didn't think to ask Dr. Palmeiro was if the fact that there was only that one parasite could be indicative as to the effectiveness of the de-parasite/worming medications you guys pointed me to earlier in this thread.

Here's a more zoomed in shot of that nematode:

If you were able to see this on the live-image of the microscope instead of this still capture, you'd be able to see that this nematode wasn't quite dead yet (still moving a little bit). In case you're curious, the mouth is on the end of the body (kind of like a worm). The histopathology should help identify exactly which type of nematode this is.

That's all for the pictures worth sharing. In my next post, I'll talk about some more of the things I learned from him about TB/_Mycobacterium_ and some of my brainstorming on what my strategy should be moving forward.


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## FlyingShawn (Mar 4, 2011)

Dr. Palmeiro seemed to have a fair amount of experience working with TB and was pretty well versed about it, so I wanted to share some of the other things I learned:

In my research, I saw some disagreement about the effectiveness of bleach against _Mycobacterium_. He was of the opinion that bleach _does_ work, but that it likely needs to be in a higher concentration than what you'd normally use (he suggested a 10:1 water/bleach mixture). He also told me how when sterilizing tanks, he often prefers to take a spray bottle and spray isopropyl alcohol around the tank instead of using bleach. His reasons were that it was easier to cover every nook and cranny with the spray bottle and because it evaporates, it doesn't require the numerous rinses you typically do after using bleach (he didn't say if he does any rinses after the alcohol or just waits for it to evaporate).

It is true that the vast majority of all tanks have _Mycobacterium_ in them at some low level. However, it isn't known what makes a fish become infected with it and it seems that certain species are more susceptible than others. Rainbows, most labyrinth breathers (gouramis, bettas, etc), and I think he mentioned some cichlids seem especially vulnerable to it. Koi, on the other hand, seem to be totally immune. He wasn't aware of any cases of Koi infected with TB, even when kept in tanks that were known to have it (I believe he meant "known to have it" as being infected with it, not just having it at the low-level background that is considered normal). So far, no one knows why some species are especially more/less susceptible.

_Mycobacteria_ seem to live primarily in the biofilm on the sides of the tank and in the filter. I also seem to recall (not positive on this one, but it stuck out in my mind) that there typically isn't a lot of it living on the substrate of the tank. There isn't much in the water itself, so UV sterilization of the water is only of limited effectiveness. Ingestion of infected matter is the main way that fish catch it.

He seemed very familiar with ongoing research in trying to treat _Mycobacterium_ with antibiotics, to the point that I got the impression that he knows some of the researchers personally. The point of their research is to treat $5000 sea horses, so there's actually potential value in using very-expensive antibiotics. So far, however, nothing has proven effective.

While we're on the subject, he also added that antibiotic baths (like the Kanamycin bath I'd been using) are so nearly useless that he likened them to the effectiveness of a person taking a bath in antibiotics. Since fish ingest so little fluid, the main way that such antibiotics would enter the system is through the gills. Medicating their food (and injection, of course) seems to be the only effective method. One of the best ways to dose them is to mix the medication into a gel-based food, so that it's easy to mix in and will stay in the food as they eat it (as opposed to soaking a dry food in medication).


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## FlyingShawn (Mar 4, 2011)

*So, now what?*

For the moment, his recommendation was to do nothing (aside from being really careful to protect myself from infection, such as not reaching into the tank with a cut and washing my hands really well after working on it), at least until the histopathology report comes back. In the meantime, he's going to research the latest data about TB and what antibiotics are considered to be the most effective. Following that, we've discussed two main options for what my strategy will be moving forward (and I came up with a third on the long drive home, so I haven't gotten any feedback on it from him yet):

*1)* Nuke everything. Euthanize all of my fish, toss the plants, tear down the tanks, and bleach/alcohol everything. This is the only way to completely eradicate the problem, but would be hugely expensive and the amount of labor required would be astronomical. 

Since I haven't been practicing great isolation techniques, I have to assume cross-contamination and this nuke would also include my smaller tanks (a 20gal, 10gal, and maybe the 1gal betta). For those tanks, the labor is much more manageable (especially since I was about to redo the 20 and retire the 10 anyway), but I'm not sure a full nuke is even doable for the 52gal. I live alone and that tank is too big for my bathtub, so I'm really not sure how I'd perform that level of cleaning (when I first got it, I was able to get someone to help me take it to a friend's house to bleach/rinse in their driveway, but that's not really an option in the winter). 

On top of that, the tank has mineralized top soil for the substrate (obtained at great difficulty from a formerly honest and reputable forum member who tried to scam me, so it's not readily replaceable). So, I'd have to extract the dirt, figure out a way to clean it without damaging it, and order new potash/dolomite to go under it when I redo the tank.

*2)* Attempt treatment. Basically, this would be built around the idea that since nearly all tanks have _Mycobacterium_, it might be possible to fight it back down to normal background levels. Even though antibiotics have not proven effective enough to cure an infected fish, there's a chance they might help a fish who either isn't yet infected or is in the very early stages. This was my idea, not his, but he admitted that it had at least a small chance. If nothing else, it might slow down the spread of the disease in the tank.

If the histopathology shows any other bacterial or parasitical problems going on, we'd also treat for those in hopes of keeping the tank as healthy as possible (and hopefully thus holding off further infections by keeping the fish's immune systems at full strength).

To do it, I'd clean the tank as well as I can while it's still running, including vacuuming up mulm and scrubbing as much biofilm as I could off the sides with an algae pad. The fish would be fed with an antibiotic medication and any fish that began to show symptoms would be immediately removed and euthanized.

On the plus side, this would be my only chance to save my livestock and even fairly expensive antibiotics would still be cheaper than a full nuke would cost me. However, the TB would never totally be eliminated from my tanks and I'd have to always treat them as being infected when working on them (gloves, thorough hand-washing, etc).

For the moment, at least, this is our primary strategy. Once we get the histopathology report and do that further research, that might change, but he made a point of not recommending drastic action just yet.

I'd really like your feedback on option 3, so I'm going to make it easy to reply to by making it a separate post...


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## FlyingShawn (Mar 4, 2011)

*3)* Nuke _in situ_ / EMP

Before I start, I should reiterate that I came up with this idea on the drive home from the vet and haven't discussed it with Dr. Palmeiro yet, so I have no idea what he'd think of it. I should also add that this is very early-stage brainstorming, so I'm really eager to get your feedback (even if you think it's a horrible idea and totally un-feasible).

If the complete teardown/sterilization strategy earlier can be likened to a "nuke," this idea is the equivalent of a biological EMP (Electromagnetic Pulse). The concept is an attempt to answer the question "Is it possible to "nuke" the tank without performing a full teardown of it?" 

At least in its current formulation, here's how it would work:


*Preparation:*

-The smaller tanks would all be given the traditional full nuke (except for the betta). Their occupants (mostly guppies) would be euthanized, plants would be treated as I'll outline below, and all substrate, decorations, equipment, and the tanks themselves would be fully sanitized (probably a two-stage method employing a 10:1 concentrations of bleach and then isopropyl alcohol). 

-My 1gal betta tank would be left running, for the time being. I got him to celebrate a special occasion a year and a half ago and have always practiced good isolation techniques in order to protect him, so I think the chances of him being infected are pretty low. Those techniques haven't been full-quarantine perfect, but his exposure has been very minimal: essentially limited to the occasional claw-tool that had been in another tank, my hands, and a couple transplanted snails that came from the 10gal (which has had potential for cross-contamination, but never shown any signs of it). I'd let him live out his life under strict quarantine and nuke the tank once he eventually dies (I don't think most bettas live too terribly long anyway)

-Depending on your feedback about a later point, the 20gal would be set up with a relatively low amount of substrate/decorations and given a fishless-cycle.


*Fish*

-All smaller/inexpensive livestock would be euthanized: Guppies, Otos, Phantom Tetras, Cardinal Tetras (sorry msjinkzd!), Neon Tetras, even the three remaining Odessa Barbs.

-Fish would be euthanized with clove oil and then placed in the trash, so as not to "share" the TB with other aquarists via flushing them.

-The larger/more expensive/more personally-valuable fish, specifically the three Rainbows, two Dwarf Gouramis, and my German Blue Ram would be placed in VERY-strict, long-term quarantine in the 20gal. The main reason I'd try to save them is that I went to great effort to obtain them (such as flying up to Albany, NY, fighting the store supervisor to sell them to me, and then having to fly all the way down to Charlotte to get back to Harrisburg due to weather) and because of that they have sentimental value.

Once in quarantine, they would be fed with antibiotic-medicated foods.
I would have separate buckets and tools designated for the tank and would practice strict quarantine methods.
If the tank is able to run for a full year without any deaths, I'd assume that although they'd been exposed, they had not become infected with the _Mycobacterium_. Since at least two, if not all three, of these species are considered "especially susceptible" to TB, I doubt they'd be able to go a full year without showing any symptoms if they are infected.
If any of them started to show symptoms, they would be immediately euthanized. On top of that, all of the others would be temporarily moved to another QT, a full nuke would be performed before returning them to the tank, and the one-year clock would reset.
NOTE: this is one of the main points I'd like your feedback about.


*Plants*

-All the plants in the main tank would be removed and treated to a 10:1 bleach dip for 1 minute or 70% isopropyl alcohol dip (haven't figured out the contact time yet), depending on if I think they'd be able to survive the bleach.
-I'm expecting that some of the plants won't survive this sort of treatment, but even if I can only save half of them, that'd still be a huge amount of money that I'd save on getting the tank set up again.


*Tank and Equipment*

-All water would be drained
-Decorations (rocks, driftwood, a couple decorations) would be removed and given either a bleach dip or an isopropyl alcohol dip (need feedback on which one, but leaning towards the alcohol since I'm not sure how bleach would affect the driftwood).
-Inside walls are scrubbed to try to remove as much biofilm as possible
-Depending on your feedback, sterilizing the rest of the tank would happen one of two ways (or a hybrid):

Entire tank is filled with a 10:1 bleach concentration, immediately drained, and then completely filled/drained another 3-4 times to rinse it out. This would be the easier method, but I'm worried about how the bleach would affect the mineralized top soil/flourite cap and if the exposure time would be too long for the silicone seals of the tank. By my estimates, the bottom of the tank would be exposed to the 10:1 mixture for 45min to 2 hours before being diluted by the first rinsing stage (best/worst case based on how long it would take to fill/drain/fill the tank).
Bottom of tank (~1" above substrate) is filled with isopropyl alcohol. Rest of tank is sprayed-down/scrubbed again with isopropyl alcohol. After maybe 15 minutes contact time, the tank would be filled/drained twice to remove the alcohol. I'm also concerned about if the alcohol would have any detrimental effects for the substrate.
-The plumbing system for the filter would be attached to a 5gal bucket containing bleach solution and run for 15 minutes.
-Fill/drain hose would be attached to faucet, used to drain a bucket of bleach solution, then used to fill/drain bucket a few times as a rinse (if I use bleach on the entire tank, this step won't be necessary).
-All other equipment and buckets would be sanitized with bleach solution (or alcohol, if I think the bleach would damage it)


*That's it, everything would be declared "clean" and I'd start the long and expensive process of rebuilding*

*The three main things I need feedback on from this idea are:*
-Does this idea of a "nuke _in situ_" sound feasible?
-Would my one-year QT idea work? (Specifically, would it be safe to call them "clean" after that long?)
-Would either a bleach or alcohol bath sterilize my mineralized top soil and flourite cap without damaging it? How about the silicone seals? Which one would be better?


*Thanks everyone for all your feedback and insight through all of this! I know these last couple posts have been lengthy, but I hope the info in them was worthwhile!*


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## DaveK (Jul 10, 2010)

First, this is an excellent set of posts on the results of the test you had done.

I'd say that there would be some risk, but you could quarantine the fish you consider valuable, and see what happens. As for the plants and substrata, I'd replace them. Plants would likely be killed by any disinfectant you used, and the substrata is just too difficult to really clean. The rest of the stuff, rock driftwood, and equipment should be ok.

Yea, it's a pain to replace substrata, but doable. Plants can be regrown. 

Good luck with this project.


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## msjinkzd (May 12, 2007)

Did you ask teh vet how long the pathogens would survive if you dried otu the tank? As in, instead of draining and nuking then refilling immediately, what if you let your tank and equipment totally dry out, while keeping the fish in a temporary tank. Even a plastic tote can be a great qt while you figure allt his stuff out.

You could then do the spray bottle with alcohol and let that dry, wipe it out, and refill.

I don't know *anything* about mineralized topsoil, so am really limited in what I can offer there. As far as sterlizing filters and equipment (nets, heaters, etc), believe it or not, I run them through my dishwasher on the hot/sterilize cycle (no soap). 

As for if they would be "clean", I think the answer is probably no. From what it sounds like, they will never really be clean. Just because the fish that have been exposed are ok, it does not mean that new additions would be as tolerant, sadly.

Also, should you decide to medicate, I am selling gel foods and would donate a starter pack for you to try, as the vet mentioned, its a great vehicle for delivering medications.


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## FreshtoSalt (Dec 1, 2011)

Excellent job at communicating your findings, thanks for sharing.

I found this thread a little while ago, doing research on the same type of problem you are having. I was hoping that it was parasites, but after this thread, I think I have a problem with Fish TB too. 

So, as far as giving you my opinion. I have a hard time pulling the trigger on the full nuke/euthanasia option. If a low percentage of your fish have been infected I would try to treat the tank with antibiotics, after euthanizing all the fish that are showing any type of symptom, and see what happens for a few months/year (this is what I am doing). So I guess I would go with option 2.
But if I see that I keep losing fish, within a month or two, I will fully nuke/euthanize the tank and start over.

Regarding your specific questions. Again, just my opinion/intuition:

-I would not try to sterilize soil/substrate.
-The one year QT process could work to keep to background levels (as previously stated).
-After this experience, if it gets to that point, I will be sterilizing with bleach, alcohol, and Calcium Hypochlorite. No more guessing...

I'm so frustrated, so much effort to do things well, and this happens. The thing is, when you get a new fish at the store you would have to quarantine it for up to a year to see if it carries TB!? a 4-6 week QT can't be trusted with TB. Are all of us playing Russian Roulette with Fish TB?

Good luck!


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## FlyingShawn (Mar 4, 2011)

DaveK said:


> First, this is an excellent set of posts on the results of the test you had done.
> 
> I'd say that there would be some risk, but you could quarantine the fish you consider valuable, and see what happens. As for the plants and substrata, I'd replace them. Plants would likely be killed by any disinfectant you used, and the substrata is just too difficult to really clean. The rest of the stuff, rock driftwood, and equipment should be ok.
> 
> ...


I already practice a bleach dip on all my new plants, since there isn't a reliable way to quarantine them. It's not as high of a concentration as I'm proposing here, but I've had good success on most plants (Hornwort being the most notable exception: it falls apart at even relatively low concentrations). I have a friend who recently used a 9:1 bleach concentration for one minute on some Ludwigia stems and a couple regular/lace Java Ferns and she's going to let me know how they do (they seemed ok immediately after being planted). She also did a 30 second dip in 70% isopropyl alcohol for some Hornwort and Java Moss and I'm waiting to hear back on the results.

If it's true that the Mycobacterium don't really live on the substrate much, I'm big as worried about getting it perfectly sterile. As long as I can drop it down to normal background levels, I'd consider it a success.

While I understand your concerns about the effectiveness and practicality of cleaning the plants and substrate, my estimates indicate that replacing all of them would roughly triple the cost of a rebuild, so I have a great deal of incentive to explore this possibility. Contrary to what a lot of people believe, we airline pilots really don't make all that much (you're lucky to make $20k your first year in the industry and it doesn't go up quickly from there), so that kind of cost is a MAJOR factor for me.


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## FlyingShawn (Mar 4, 2011)

msjinkzd said:


> Did you ask teh vet how long the pathogens would survive if you dried otu the tank? As in, instead of draining and nuking then refilling immediately, what if you let your tank and equipment totally dry out, while keeping the fish in a temporary tank. Even a plastic tote can be a great qt while you figure allt his stuff out.
> 
> You could then do the spray bottle with alcohol and let that dry, wipe it out, and refill.
> 
> ...


Excellent question, Rachel! I don't know why I didn't think to ask him about drying it out, maybe it seemed like it'd be too simple. That's now my top question for when I talk to him next.

You know, that dishwasher idea is also really good. I'll have to start using that for a lot of my smaller equipment!

Perhaps "clean" wasn't the best choice of words for what I was going for with the long-term QT idea: maybe "as clean as anyone else"?. It centers around the idea that TB exists at some low level in all tanks, so if the antibiotic is effective enough to beat it back down to that level in not-yet-sick fish, they wouldn't be any more hazard for the rebuilt tank than the replacement fish I'd be getting. I figure that since the two (or maybe all three) of the species listed are considered especially susceptible to TB, they're going to get it within a year if they're going to get it at all. I wouldn't medicate them for the full year, probably just the first month (or whatever is recommended for the specific antibiotic) to give them the opportunity to come down with symptoms if it wasn't effective.


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## FlyingShawn (Mar 4, 2011)

FreshtoSalt said:


> Excellent job at communicating your findings, thanks for sharing.
> 
> I found this thread a little while ago, doing research on the same type of problem you are having. I was hoping that it was parasites, but after this thread, I think I have a problem with Fish TB too.
> 
> ...


Good luck with your fish, Fresh! I certainly know that this is a VERY frustrating experience. My initial plan was to go for Option 2, but as all this has sunk in over the last few days I've begun to wonder if some variety of nuke is somewhat inevitable simply on the basis of having to protect my own health. I just don't know how long I'd be diligent about good quarantine procedures and if I was, if it would cause me to burn out on the hobby in the long run. On top of it all, I'd feel bad introducing any new fish to the tank, which wasn't even fully stocked yet, because it'd potentially be a death sentence!

That's not to say that I've ruled out Option 2 yet, I just wanted to share my thought process for what's driving me to consider all the options as thoroughly as possible.

I think I've talked about most of your points in the last couple replies with others, but I do want to talk about your last one for a minute. The reason I'm looking at a one year QT for those guys is because those specific species are known to be especially vulnerable and because I positively know they've been exposed to it.

When it comes to QTing new arrivals, I'm also dealing with the same questions you're asking. I don't think a year would be necessary. A couple months ago I started practicing a 1 month QT, prior to that (when I got these guys) I was only using a week. Now I'm considering 1.5-2 months and might go as high as 3. It's certainly possible for TB to elude you longer than that, but there has to be a point of "acceptable risk" and I'm guessing most fish will begin to show symptoms within that time frame if they have it. Not an expert opinion, of course, but that's the direction I'm leaning.

Please let us know what you decide to do, both for your tanks and QT procedures!


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## esteroali (Oct 24, 2007)

*Uv uv uv !!!*

Prolonged sunlight is the best method of sterilization. UV in tank filters are also somewhat effective. Bleach is useless as is isopropyl alcohol IMO. I have TB is a 20 G tank, verified by necropsy in fish and myself. I exercise caution in this tank. The tank has no ill fish now,I do believe personally it is species specific as far as susceptibility. I did not have the intestinal fortitude to totally break down this tank and I am careful not to trade any specimens from this tank.


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## FlyingShawn (Mar 4, 2011)

esteroali said:


> Prolonged sunlight is the best method of sterilization. UV in tank filters are also somewhat effective. *Bleach is useless as is isopropyl alcohol IMO.* I have TB is a 20 G tank, verified by necropsy in fish and myself. I exercise caution in this tank. The tank has no ill fish now,I do believe personally it is species specific as far as susceptibility. I did not have the intestinal fortitude to totally break down this tank and I am careful not to trade any specimens from this tank.


The implications of the line I highlighted are, to say the least,_ troubling_ for someone in my situation. Since I don't have a high powered UV wand and no single place in my apartment gets sunlight for more than a few minutes each day, this would rule out just about any chance I'd have for eliminating this disease!

I'll be honest with you, at this point I'm not sure what to believe. On the one hand, I have Dr. Palmeiro recommending bleach and alcohol based on his research (and possibly knowing people who are doing active research on it), while on the other hand I have you and your personal experience with TB saying otherwise. If you search the net, you'll find a full spectrum of opinions ranging from "just use bleach, that'll kill anything" to "nothing works! Myco can even repair it's own RNA/DNA so it's even UV-proof!"

Ever since your post, I've spent the bulk of my free time in-between flights and in the hotel actively searching the net in hopes of finding some definitive, scholarly research. What I found may not be definitive, but they certainly ranks among the most scholarly works I've seen on the topic:

Here's a fantastic article by Diana Walstad: Mycobacteriosis− the Stealth Disease.
She suggests that repeated, routine disinfections can actually _increase_ the population of _Mycobacteria_ in a tank! However, that's not to say that she suggests _never_ chemically nuking a tank, in fact it may be warranted when dealing with particularly large outbreaks, just that numerous, repeated nukes could be detrimental. She also strongly recommends UV sterilizers on an infected tank and suggests that TB can actually be beaten back through providing an environment where faster-growing "normal" tank bacteria can out-compete MB for resources! For my specific case, I'm not sure if the take-away of this article is more in favor of one of the nuking ideas or "Option 2", I'll need to re-read it a few more times and think about it. *Side note for FreshtoSalt*: read her QT recommendations, it seems like 2-3 months is a good target.


This article from the University of Florida Extension: Institute of Food and Agricultural Sciences has the following in its "Disinfection" section:


> Mycobacteria are resistant to many commonly used bactericidal agents at standard dosage rates, including chlorine bleach and quaternary ammonium compounds. As much as 10,000 ppm chlorine has been reported necessary to kill mycobacteria. Mycobacteria are sensitive to 60-85% alcohol. In addition, there are special products marketed specifically to decontaminate mycobacteria-infected surfaces. However, if these are used, efforts must be made to remove all residues of disinfectant.



The last link for tonight is from Adrian Tappin's *free* book Rainbowfishes ~ Their Care and Breeding in Captivity, which I've heard referred to as the "definitive work" on the subject. The link is to the relevant section in an _ugly_ web-based version of the book, but you can also download the 200MB(!) book-formatted PDF version on that site. Here's the last paragraph of his "Treatment" sub-section:


> Veterinarians at the National Aquarium in Baltimore, USA recommend using chlorine to clean the tank and substrate, etc., and then spray 65-90% isopropyl alcohol onto the glass, and allow it to dry. They recommend the alcohol as they found that chlorine does not kill all mycobacteria. They use chlorine to remove/oxidise organic material to assure the alcohol contacts all mycobacteria in/on the tank. Remove all residues of disinfectant from the aquarium before reuse. _(Denise Petty DVM, pers. comm. 1998)._



*Based on all this research, here's my tentative conclusion for the night:*

esteroali is at least partially right: bleach is of very-limited effectiveness against _Mycobacteria_ when using concentrations and contact times that wouldn't destroy the aquarium through silicone-seal damage. However, my research couldn't find any reliable backing for his assertion that alcohol is also useless (if anything, the sources I found suggested the exact opposite). He could still be right on that point, I just didn't find it. Given the lack of actual data, I'm not sure there's any way to confidently know for sure, short of actually running the laboratory studies to test alcohol-alone and bleach-then-alcohol protocols. In the end, whatever I do will be as much an "educated guess" as anything else.

At this point, I'd probably lean towards an "Option 3" EMP nuke inspired by the National Aquarium bleach/alcohol "one-two punch" protocol in that last quote. The idea would be to use the bleach to begin the killing process and break down the biofilm in the tank, then use the alcohol to finish it off.

Farther down the line, I'd continue to run UV on the rebuilt-tank and likely get an additional UV unit to use on the long-term QT for the fish I'm trying to save. I'll also probably employ a lot of Diana's quarantine recommendations in the future (especially the use of "Sentinel Fish").

What do you guys think?


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## FlyingShawn (Mar 4, 2011)

What about ultra-high concentrations of salt?

I know _Mycobacteria_ is just as dangerous to marine fish as freshwater, but I'm talking like Dead Sea near-saturation level salt. Would it be able to survive 24 hours in that?

If it worked, I could nuke the filters, tank, substrate, and decorations all at once without having to worry about damaging the seals of the tank. After a few big drain/fill cycles, the tank would be once again safe for occupancy without fear of toxic residues.


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## FlyingShawn (Mar 4, 2011)

I emailed Dr. Palmeiro about some of these questions today, here is what he said:

When asked about how much _Mycobacterium_ resides in the substrate (because I seemed to remember him saying there wasn't as much to be found there):


> I said it concentrates more in the biofilm, it is definitely found in the substrate as well, especially with higher organic loads


With regard to the idea of soaking the substrate in alcohol for ~30min to sanitize it in the tank:


> Bleach followed by alcohol would probably be better, but this would probably work. If possible, i would STRONGLY recommend replacing the substrate.


In response to Msjinkzd's idea of drying out the tank for a couple months:


> Mycobacteria are fairly resistant to dessication, I would not rely on this as a method for disinfection


And finally, in response to my idea of trying ultra-high salt concentrations:


> Salinity alone is not enough to kill the bacteria.


I also found this thread about sanitizing Flourite on the SeaChem support forums:
Cleaning/Disinfecting Black Flourite


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## FreshtoSalt (Dec 1, 2011)

I finally had some time to digest Diana's article and wanted to comment on a couple things.
Once I realized that indeed is probably FTB, the background level concept was what my intuition was telling me made the most sense. Due to how hard it is to completely eradicate these bacteria, how frequent they are in aquariums, and for the following reasons. 

In my case, I acquired a 55g with two fish from a friend. It was fully established (5 yrs or so) and after giving the fish away (cichlids) I started a community tank. Since the tank was cycled I stocked it fairly quickly, within 3-4 months I had most of the fish I wanted in there. 
Except some neons that died after a month in the tank, none of the other fish seemed ill. Water parameters never got out of whack, and I only had one Ich out brake for the next 5 months. Then, one day one of my stigmata loaches developed a curved spine and started to lose weight, another (out of 4) started swimming erratically. Took both fish out, euthanized one and the other died in a hospital tank. That is when I started doing research, and concluded that it was either parasites or Fish TB. So I treated for parasites.
For 2-3 months after that, everything was fine so I acquired 4 GBR's. Again, for another 4-5 months everything went well until one day, with NO symptoms (that I could tell), one of my paired/breeding angel fish died, I didn't understand, maybe I was just overstocked (~20+ growing fish).
Then I moved out of State. All the fish made the 900mi. drive, but after 2-3 weeks the GBR's started dropping like flies and my betta developed the bloated stomach with wasted body sintoms. The GBR's all got progressively worse hollow bellys and after a couple weeks they all passed, while the betta was just hanging in there. I thought for sure parasites, and maybe the new water weakened the fish, while TB was in the back of my mind. So I treated for parasites with quick cure, and after a month I acquired 4 new fish, they all died within 2 weeks, but some had stringy feces, so I kept thinking maybe it is hard core parasites...
At this point I decided that I am not going to get any new fish, upgrade to a 120g to give a better environment to the rest of my stock (~20 fish now), nuked the 55g, and made it bare bottom with decorations to keep as hospital tank until the 120g cycled. I treated with Flub. for parasites and waited, soon enough, I got a nitrite spike on the 55g and lost 2 more fish despite my efforts with WCs, and the betta was so wasted I euthanized him. 
So after a couple more weeks of daily WC and a lot of observation everything was well. I started introducing the fish from the 55g, that looked healthy and never displayed any symptoms whatsoever, into the 120g. At this point, only the fish that had been completely healthy for the 12+ months that I've had them were transferred to the 120g. 
Then I found your thread and the TB diagnose, kicked myself in the head, since I have now introduced "healthy" carriers of the disease to my new tank (potentially) and I am pretty sure it is TB. That is why Option 2 is what I am doing now. I will keep transferring the fish in the 55g little by little to the 120g. 

Conclusion: Basically, I've had a contaminated aquarium for probably almost 1 year now, and most of my fish, about 80%, seem just carriers/survivors. I believe that except in two occasions were deaths happened for "no reason", the flares were always months apart and due to some concrete environmental change in combination with overstocking; and also when new fish with no immunity were introduced.

So, since I am moving again in 6 months, I am going to commit to not getting any new fish, buying a UV sterilizer and see what happens. If the fish keep getting sick I will euthanize right away.
After I get established at a new place, and if my fish survive for another few months, I will seriously consider the "Sentinel" idea for my QT tank and a 2-3 month QT for any new additions. If I lose all my stock, I will sanitize the tank as much as I can (probably with a combo of different sanitizing agents) and turn the 120g into a salt water tank, and do something different with the hobby in order to keep my sanity.

(Sorry for the long post, English is my second language and I was trying to explain everything without confusing people)


FlyingShawn said:


> [...] My initial plan was to go for Option 2, but as all this has sunk in over the last few days I've begun to wonder if some variety of nuke is somewhat inevitable simply on the basis of having to protect my own health. [...] On top of it all, I'd feel bad introducing any new fish to the tank, which wasn't even fully stocked yet, because it'd potentially be a death sentence!
> 
> That's not to say that I've ruled out Option 2 yet, [...]
> 
> ...





FlyingShawn said:


> *Side note for FreshtoSalt*: read her QT recommendations, it seems like 2-3 months is a good target.


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## Rich Conley (Jun 10, 2008)

DaveK said:


> First, this is an excellent set of posts on the results of the test you had done.
> 
> I'd say that there would be some risk, but you could quarantine the fish you consider valuable, and see what happens. As for the plants and substrata, I'd replace them. Plants would likely be killed by any disinfectant you used, and the substrata is just too difficult to really clean. The rest of the stuff, rock driftwood, and equipment should be ok.
> 
> ...


 
I'm gonna have to disagree here.

If you're going to quarantine and keep some of the fish, there's no reason to not keep the plants with them. The fish are the main risk here. The fish are the vector. If you're keeping the fish, you might as well keep as much as you can.



I've had a lot of conversations with people about Myco, and its pretty tough to get rid of. Its something that pretty much every major aquarium out there just kind of lives with. 

Employees of these places regularly fail TB tests because of myco.


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## FlyingShawn (Mar 4, 2011)

FreshtoSalt said:


> Conclusion: Basically, I've had a contaminated aquarium for probably almost 1 year now, and most of my fish, about 80%, seem just carriers/survivors. I believe that except in two occasions were deaths happened for "no reason", the flares were always months apart and due to some concrete environmental change in combination with overstocking; and also when new fish with no immunity were introduced.
> 
> So, since I am moving again in 6 months, I am going to commit to not getting any new fish, buying a UV sterilizer and see what happens. If the fish keep getting sick I will euthanize right away.
> After I get established at a new place, and if my fish survive for another few months, I will seriously consider the "Sentinel" idea for my QT tank and a 2-3 month QT for any new additions. If I lose all my stock, I will sanitize the tank as much as I can (probably with a combo of different sanitizing agents) and turn the 120g into a salt water tank, and do something different with the hobby in order to keep my sanity.


Thanks for sharing the story and what you're thinking!

I'm also seriously considering the "Sentinel" idea. From the literature I've been reading, I've gotten the impression that Zebrafish are especially susceptible to TB, so I'm thinking of getting some of them specifically for the purpose of becoming Sentinels.



FreshtoSalt said:


> (Sorry for the long post, English is my second language and I was trying to explain everything without confusing people)


I completely understand! Even though English is my first language, I tend to write extra-lengthy posts for the same reason!


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## FlyingShawn (Mar 4, 2011)

Rich Conley said:


> I'm gonna have to disagree here.
> 
> If you're going to quarantine and keep some of the fish, there's no reason to not keep the plants with them. The fish are the main risk here. The fish are the vector. If you're keeping the fish, you might as well keep as much as you can.
> 
> ...


Thanks Rich.

For the plants, my real question is if _Myco_ remains on the surface of the plants or somehow gets into the plants themselves. If it remains on the surface, I should be able to sterilize the plant chemically and have it be safe to introduce into the new "clean" (non-QT) tank. If it somehow gets into the tissues of the plant (I have no idea how that would happen, but I'm trying to consider every possible vector), then it could be immune from the sterilization procedure and would have to be discarded.

Whether the plant survives the sterilization procedure is another question entirely, but I'd rather try and see what happens than just throw it away. If I determine the plants are worth trying to sterilize, I'll be setting them up in an emersed-growth rig to keep them alive while I go through the whole "nuke" process on the tanks.


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## FlyingShawn (Mar 4, 2011)

Dr. Palmeiro emailed me a couple of factsheets on _Mycobacterium_:


> I think you had the one from Univ of Florida. The one from SRAC is new and has some of the more recent papers summarized in it.


The first one was the University of Florida one I linked to earlier, the second one can be found here:
SRAC Mycobacterial Infections of Fish

I'm in the process of picking his mind about bleach concentrations and contact times, as well as the possibility of heat-killing the bacteria (such as baking/boiling the substrate and driftwood as a non-chemical or secondary kill method). I'll let you know what I find out when we get a little farther along in the discussion.


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## FlyingShawn (Mar 4, 2011)

The histopathology results are in, here's how the vet summarized them:


> Results were compatible with mycobacterium. The GI tracts looked normal with no evidence of parasites. One fish also had a gonadal tumor, but this was likely an incidental finding and not related to the clinical disease. It was a severe case with numerous granulomas, mostly concentrated in the kidneys.


As far as bleach concentrations:


> <Me> Speaking of the bleach stage, if I use a 10:1 water/bleach concentration (or maybe 9:1 for ease of measurement), what would you recommend for the contact time?
> 
> <Vet> According to the variable papers out there, concentrations upwards of 50000 mg/L may be required. With 5% bleach (50 mg/ml), you would need to add 1000 mls into 1 liter to achieve the necessary concentrations, which equals 50/50 dilution. Bottom line, I am ok with 1:9, but you need to do it for about 24 hours to try to get a prolonged contact time to kill the bacteria


I've asked him about contact times for the higher 50/50 dilution, but have gotten no response. The bigger question on my mind is if the silicone seals on the tank could survive a 1:9 concentration for 24 hours! I've heard anecdotal stories that bleach will damage the seals, but never seen anything concrete on the matter.

The other question is if such a contact time is even necessary. As he said, it would be required to _kill_ the Myco, but if I'm doing the Lysol/Alcohol stages later, don't I really only need the bleach to break down the biofilm to prepare for those other chemicals?

I've also asked about using boiling or baking to heat-sanitize driftwood/rocks/substrate/etc, but he says he has "no clue" as to the answer. I've found some scientific literature that might seem to indicate it would work, but don't have the background to know that I'm interpreting it correctly. To get some fresh input on the matter, I've posted a thread in the "General Planted Tank Discussion" forum: Anyone with biology knowledge: boiling or baking to kill Mycobacteria/Fish TB?


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## unissuh (Jun 5, 2006)

I haven't read this entire thread, but I think you're starting to split hairs here - keep it simple. Going through theories of what is the most effective is only useful to a point, you just want something that is good enough to disinfect the system, it doesn't need to be perfect enough to kill everything on the planet.

It's not necessary to go to such a high bleach concentration, the point of the bleach isn't to kill the Mycobacteria. If you're cleaning out the filters and so forth, 1/10 for a short period, less than an hour will probably be fine. Like you said, the point is to strip the biofilm and other organics. If you're following it up with a good dose of 80% alcohol, that is the part that is designed to kill the bacteria. What is probably more important is to give everything a good scrub before and after the bleach dose to try and remove as much organics and crud as possible. The bleach is only the backup to melt all the little bits that you miss. 

Heat will probably knock it off - marinum is a low temp bug, not as temp tolerant as some others in the genus AFAIK. Boil it for a while and it will probably be fine. Wood and rocks are fine with this or soaking in alcohol, I'd ditch your substrate and plants or keep them in a known contaminated system because I don't think it's worth the effort of disinfecting to a satisfactory degree.

Quite frankly, most people are not set up to prevent Mycobacterium entering their systems (including me), and I wouldn't even bother trying. The main issue is how on earth do you tell when a fish has Myco (or other pathogens for that matter)? Acute infections and disease is one thing, but diagnosing a carrier is flat out impossible without euthanising several in a group and performing histology or molecular based diagnosis. Even in this case, the group would have to be held for a long time before the diagnosis to make sure any possible disease is transmitted. You can knock out most parasites by throwing praziquantel, levamisole, fenbendazole etc at them, but try doing this for antimicrobials and you'll probably end up with all sorts of resistant bugs growing in your quarantine tank over time.

The only way I can think of to make sure your stock aren't carriers is to allocate tanks into "clean" and "dirty" facilities like many scientific animal houses are set up. Trust no one & only transfer disinfected eggs into your clean facility. Even this is not 100% without some good screening, but it's as good as you can get without some specialized equipment and/or reagents.


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## FlyingShawn (Mar 4, 2011)

unissuh said:


> It's not necessary to go to such a high bleach concentration, the point of the bleach isn't to kill the Mycobacteria. If you're cleaning out the filters and so forth, 1/10 for a short period, less than an hour will probably be fine. Like you said, the point is to strip the biofilm and other organics. If you're following it up with a good dose of 80% alcohol, that is the part that is designed to kill the bacteria. What is probably more important is to give everything a good scrub before and after the bleach dose to try and remove as much organics and crud as possible. The bleach is only the backup to melt all the little bits that you miss.


That's roughly what I was thinking. I'm not sure why the vet was recommending the long contact-time when the other chemicals are the "kill" elements.



unissuh said:


> Heat will probably knock it off - *marinum is a low temp bug, not as temp tolerant as some others in the genus AFAIK*. Boil it for a while and it will probably be fine. Wood and rocks are fine with this or soaking in alcohol, I'd ditch your substrate and plants or keep them in a known contaminated system because I don't think it's worth the effort of disinfecting to a satisfactory degree.


I've bolded the key part here. My guess is the same as yours, but I'm really hoping to find someone who can actually say with some authority that this is right (hence why I started the other thread I linked to).

I'll be posting the latest set of plans in the near future, but they essentially consist of a "full nuke" of the existing tanks, with the most valuable fish being moved into a "dirty" (to use your term) permanent QT with high-powered UV. I'll probably start the mass euthanasia tomorrow.

Thanks unissuh!


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## Jadenlea (Sep 15, 2011)

I am sympathetic to your plight here! I too have been worried about TB in my tank. 

I have a beautiful healthy LOOKING planted tank however I have had fish dying with these symptoms also. They start out first separating from their school and just sitting around in one place in the tank. Then they just seem to waste away slloowwllyyyy. They seem to linger for weeks and I just can't bear to kill them hoping they will get better. By the time they are ready to die a couple of them have twisted looking bodies. 

The strange thing is, that this has only happened to the glofish and guppies. 

I have a german blue ram, 2 balloon rams, otos, neons and a redlined shark. Oh and a male and female betta. (Yes they actually live in an uneasy truce together in the tank but it is heavily planted) None of these fish are showing any of the symptoms even though some of them have been in there longer then some of the glofish were. Every glowfish (8) has now died. The guppies died first and fairly quickly after I got them and I suspect were the ones who carried this to my tank. 

I can't bear the thought of just killing all these fish and my beautiful healthy plants so I am watching your thread carefully. It will be interesting to see if now that the last glofish has died if it will affect a different kind of fish now. 


Anyway, the thing that made me post is listening to all your plans for sterilizing your tank. I think you said it was a 55 gal. and expense is a consideration. If you are really thinking about taking it all apart...

My question is.. you do realize that if you just have one of the plain old retangle 55s, that you can toss the one you have and buy a new one at petco right? Their rectangle tanks are 1 dollar per gallon. It seems like it would be worth the 55 dollars to avoid trying to sterilize and possibly damage the seals. The rest of the stuff is smaller and easier to clean but man.. imagine if you damage the seal and refill it and it breaks. Ugh. 

If you have one of the irregular shaped tanks then I guess you are stuck...the petco sale doesnt include their corner, bowfront or other strange shaped tanks. 

Unfortunately for me, mine is a bowfront so it wouldn't work for me


PS. I work in a veterinary hospital.. I wonder how psycho one of the vets would be if I asked him if he would be interested in performing a necropsy on a fish lol. I can just picture his face now.


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## unissuh (Jun 5, 2006)

FlyingShawn said:


> unissuh said:
> 
> 
> > Heat will probably knock it off - *marinum is a low temp bug, not as temp tolerant as some others in the genus AFAIK.*
> ...


Sorry, should have probably qualified that statement a little more. It is more of a fairly well educated guess, not an assumption, but there is no single report which has tested boiling. 

What has been done is a number of studies examining heat treatment of paratuberculosis, avium etc as these are animal Myco that could be contaminating dairy products. Pasturisation (145-160F) for 20-40 min kills all of those, see no reason why boiling for the same amount of time wouldn't work.

The only "assumption" part is that marinum is as heat susceptible as other Mycos which is pretty much a certainty considering that i) it is well known for usually only infecting human hands or other extremities that aren't maintained at core body temp and ii) that one old fashioned way of treating marinum infections (that actually cured the infection alone in some cases) was heat therapy.


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## FlyingShawn (Mar 4, 2011)

Jadenlea said:


> I have a beautiful healthy LOOKING planted tank however I have had fish dying with these symptoms also. They start out first separating from their school and just sitting around in one place in the tank. Then they just seem to waste away slloowwllyyyy. They seem to linger for weeks and I just can't bear to kill them hoping they will get better. By the time they are ready to die a couple of them have twisted looking bodies.


I'm sorry to say it, but those symptoms sound exactly like what I experienced. Different species, of course, but the symptoms seem the same. Doing the necropsy was expensive, but I definitely felt it was worthwhile for knowing what I'm up against. Otherwise, I'd feel much more like you: not able/willing to take really drastic action out of any slim chance that it's not TB and the hope you can still save them.



Jadenlea said:


> If you have one of the irregular shaped tanks then I guess you are stuck...the petco sale doesnt include their corner, bowfront or other strange shaped tanks.
> 
> Unfortunately for me, mine is a bowfront so it wouldn't work for me.


Nor me, unfortunately. The tank is a 52gal Flat-Back Hex. At times like this, I really wish I'd gotten a standard 55 instead, but at the same time I really like the look of the tank and it works really well in the room. 



Jadenlea said:


> PS. I work in a veterinary hospital.. I wonder how psycho one of the vets would be if I asked him if he would be interested in performing a necropsy on a fish lol. I can just picture his face now.


Just imagine if you waited until April 1st to ask him! You'd have that much harder a job of trying to convince him you're serious!

If you have a good relationship with one of the vets and get the chance, could you please pick his brain on heat-sterilization? We've got a good discussion going on that other thread I started (Anyone with biology knowledge: boiling or baking to kill Mycobacteria/Fish TB?), but the main question seems to be centering around if a "dry heat" process can kill bacterial endospores.

Good luck with your tank, I really feel for you. Please let us know what you decide to do!


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## FlyingShawn (Mar 4, 2011)

unissuh said:


> What has been done is a number of studies examining heat treatment of paratuberculosis, avium etc as these are animal Myco that could be contaminating dairy products. Pasturisation (145-160F) for 20-40 min kills all of those, see no reason why boiling for the same amount of time wouldn't work.
> 
> The only "assumption" part is that marinum is as heat susceptible as other Mycos which is pretty much a certainty considering that i) it is well known for usually only infecting human hands or other extremities that aren't maintained at core body temp and ii) that one old fashioned way of treating marinum infections (that actually cured the infection alone in some cases) was heat therapy.


Exactly the same assumption I've been going off of in my research. There are some sources that say that Myco is spore-forming, so I'm currently leaning towards a dry-heat baking process in the oven to account for that possibility. I haven't found anything that would suggest that they can withstand 450F temps in the oven, even in the spore stage, but am still researching it.

Thanks for the input!


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## mmccarthy781 (Jul 7, 2011)

How fast does fish tb kill? I had a betta fish die two days ago and he was only sick for 1 day before that, im trying to rule fish TB out


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## FlyingShawn (Mar 4, 2011)

mmccarthy781 said:


> How fast does fish tb kill? I had a betta fish die two days ago and he was only sick for 1 day before that, im trying to rule fish TB out


It seems that most cases are extremely slow, but TB is one of those weird diseases that don't always exhibit the same symptoms. It isn't very common (the bacteria itself is, but fish actually getting sick from it isn't), so I'd probably assume that it isn't TB until you rule everything else out.


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## FlyingShawn (Mar 4, 2011)

Do you have ANY idea how hard it is to net Cardinal Tetras in a tank this big?!

Sorry, just had to vent, back to work...


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## FreshtoSalt (Dec 1, 2011)

lol, I feel your pain!
Sorry, not funny... good luck.


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## FlyingShawn (Mar 4, 2011)

At the risk of sounding cold, I guess I have to take what little humor I can find in this situation.

I've caught nearly all the fish I'm going to euthanize today (I'm sure there are more than a few tiny guppy fry who escaped my net, but otherwise I think I got everyone) and I'm now waiting for the first dose of clove oil to take effect. I'm using the clove oil + vodka method outlined here: What Is the Most Humane Way to Euthanize a Fish?

I found one cardinal that was definitely showing the emaciation associated with TB and another who might have been in the earlier stages of it. I haven't been able to look closely at the guppies (too many), but haven't noticed any looking unhealthy.


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## FlyingShawn (Mar 4, 2011)

Stage 1 is over: I've now euthanized the bulk of my fish.

I collected them in a 2.5gal and took this picture right before I began to add the clove oil:


The final kill count:
1 Red Flame Dwarf Gourami
2 Ghost Shrimp
2 Black Phantom Tetras
2 Neon Tetras
23 Cardinal Tetras
216 Guppies

I didn't notice any fish with noticeable TB symptoms other than the one Cardinal I mentioned earlier. Just to be safe though, I put them in a ziploc and tossed them in the trash instead of flushing them back into the water supply.

I've decided to keep the three Odessa Barbs and the three Oto Cats until the 20gal Permanent QT is ready and I tear down the main tank. They look healthy at the moment, so I didn't feel the need to put them down just yet. After that, I'll be left with the Betta, three Rainbows, Neon Blue Dwarf Gourami, and the German Blue Ram.

The next step will be the teardown and nuke of the smaller tanks, which I'll have to do progressively over the next few weeks due to time and workspace constraints.

I know the vast majority of the work on this project is still ahead of me, but I think tonight was one of the hardest steps. I have to admit, I think the clove oil and vodka technique did make euthanizing them a little easier, since you add a little clove oil at a time to peacefully put them to sleep and by the time you add the vodka, they don't spasm/twitch/react in any way. In the past I've used the dunk-in-ice-water method to near-instantaneously shock kill their nervous system, but there's still that second or two of violent thrashing right before they go.


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## FreshtoSalt (Dec 1, 2011)

It sucks you had to do this. I've felt like crap when I had to euthanize fish a couple times before. 
You are doing the right thing though. Including all the research that is now consolidated in this thread. 
Thank you!


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## Jadenlea (Sep 15, 2011)

I noticed ghost shrimp in the euthanize list. Does TB effect shrimp also?


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## FlyingShawn (Mar 4, 2011)

Jadenlea said:


> I noticed ghost shrimp in the euthanize list. Does TB effect shrimp also?


I don't know. I strongly doubt it, but I only had two and didn't want to take any risk of them being carriers. Interestingly, they didn't seem to be affected at all by the clove oil and actually were active for a while after I put in the vodka.


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## unissuh (Jun 5, 2006)

Yes, there are reports of shrimp granulomas.


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## FlyingShawn (Mar 4, 2011)

I think it's about time I posted an update. I have some bad news, some good news, and a general strategy/progress report. So, let's get started...

First off, the bad news:

If you'll recall from earlier, the "chemical" side of my cleaning strategy was to be a high-concentration bleach soak, followed by Lysol, and then finished with isopropyl alcohol. I devised that plan according to the information from my vet and the articles I posted earlier. However, I've hit a bit of a snag in the plan. I was having trouble finding a Lysol product with the active ingredient those articles listed ("1% benzyl-4-chlorophenol/phenylphenol"), so I ended up contacting its manufacturer Reckitt Benckiser in order to find the item I needed. After a VERY frustrating series of back-and-forth emails and being directed in circles around the company of people unable or unwilling to put any effort into answering my question (seriously, is incompetency considered a marketable skill now?!), I finally got an answer: in order to unify their product line across all 50 states and their assorted safety regulations, they no longer manufacture any products with the ingredient we're looking for. So, dead end there.

The good news is that I may not need it after all. During my hours of research, I stumbled across this scholarly article:
Efficacy of Common Disinfectants against _Mycobacterium marinum_ in the Journal of Aquatic Animal Health.

The article is paid-access-only for non-subscribers, but I lucked-out in the sense that my brother is a university librarian and has access to it through their system. Based on what I learned reading it, I'm guessing that this article is the primary source used by the other articles I linked to earlier in describing what does and doesn't work against _Myco_. As a result, there weren't many surprises in what I found, except for one fairly significant one:

Bleach _can_ work, even at less-than crazy concentrations.

There are some caveats however: namely that it requires much longer contact times (at least an hour) and, more importantly, this is _not_ a "real world" experiment, so we have to be careful what we try to apply to our tanks. What do I mean by that? These tests were done against pure laboratory samples of _Myco_, not against those hiding in the biofilm that you'd find in your tank. I've heard concerns that such biofilm could serve as a protective shield keeping the bleach from ever coming into contact with the _Myco_, which would explain the generally accepted belief that bleach is ineffective against it.

The article also said that 70% isopropyl alcohol required only a minute contact time to effectively kill _Myco_ (again, in a non-biofilm laboratory setting).

*What does this mean for me and my cleaning protocol?*

Obviously, Lysol is out. It wouldn't surprise me if the current formulation of it was effective, but at the same time I don't know that to be true, so I'm going to assume that it isn't and skip it altogether.

Here's my current step-by-step protocol for each tank:
1) Drain all water and remove EVERYTHING (decorations, equipment, substrate, etc)
2) Thoroughly scrub the tank with a "Dobie" non-scratch scrubbing pad (they're made by 3M and are amazing) in order to remove as much algae and biofilm as possible.
3) Soak the tank for 1-2 hours in a 1:9 bleach solution to start killing the _Myco_ and kill/break-down the remaining biofilm.
4) Drain about half of the bleach solution and re-scrub the entire tank again with the Dobie pad
5) Remove the rest of the bleach solution and rinse the tank with tap water
6) Set the tank out to dry for a couple of days
7) Spray the entire interior and exterior of the tank with a liberal amount of 70% isopropyl alcohol (with an extra focus on seams and frame edges) in a well-ventilated room and leave it to evaporate completely. I repeat this step at least three times to ensure that every surface has sufficient contact time for the alcohol.
8) Rinse the tank again to remove any potential residue from the alcohol (I don't think there is any, but it can't hurt to be sure).

I'm also applying adapted versions of the above process to the non-tank items that I intend to salvage. For example, I can't realistically scrub the gravel I'm saving from some of the smaller tanks, but instead I'm using much longer bleach contact times and actively stirring it during that soak in order to use the naturally-abrasive nature of the gravel to scrub for me. Also, instead of spraying the gravel with alcohol, I'll be placing it in a bucket and soaking it for an extended period of time instead (probably at least a half hour).

For the things I'm afraid to treat chemically (primarily the driftwood and maybe the lava rocks), I'll likely be using a 3+ hour bake at 450+ to heat-sterilize them. I'm still going back and forth on if that's how I want to proceed with those items, so for the moment I'm just setting them aside while I work on other steps of the project (I continue to welcome your input on this!)

I've decided to not attempt to save the plants. As much as I cringe at the idea of replacing all of them, I simply haven't been able to identify a way to ensure that they wouldn't just act as carriers for the disease. That said, however, I won't be discarding all of them right away. Contrary to my earlier plans, I've decided that the 20gal Permanent QT that will be housing my remaining fish will be a planted tank rather than a bare one, so I'll be using plants from the main tank to set up the QT (with a bleach-dip in the middle) instead of just trashing them all. The original plan was that if one of them got sick, I'd euthanize it and re-nuke that QT to minimize exposure for the others. Instead, the new plan is to still remove/euthanize the sick fish, but I'll rely on the UV unit in the tank and Diana Walstad's idea of "good bacteria out-competing _Myco_" in a healthy tank. Will this plan increase the risk of the other QT fish getting infected? Yes, however I'm not so sure that that's a bad thing, since we're already assuming they're carriers and I do have to let them die eventually so I can sterilize that tank and finally be rid of this stuff.

*So, where am I in this whole process?*

Overall, I'd guess that I'm at about 30-35% of the total labor involved (not counting wait-times, like when I'm waiting for the QT to cycle before I move fish and teardown the 52gal). More specifically:

-One of the 10gal tanks and one of the 2.5gal tanks are done with Step 7 of the cleaning protocol.

-The 20gal is almost done with Step 7 (still need to do the final alcohol spray).

-The equipment/decorations/gravel from those tanks are all done with Step 6 and are being prepared for Step 7 (since these chemicals aren't all that cheap on this kind of scale, I'm trying to group things together to get as much mileage out of each bottle as possible)

-The planted 10gal that all those guppies came from is currently in the middle of Step 1 (teardown/basic cleaning).

-The final 2.5gal has not yet been touched. I'm trying to figure out what I did wrong with my dry-start method HC carpet that seems to be failing, so I wanted to give it some more time to "rebound" just to see what would happen before I tore it down.

-My new-and-improved emersed-growth rig is now up and running (using both a larger container and a hybrid of artificial and natural light). I never tore down the original one that I was using to grow HC to start the big tank, so I have some HC, narrow-leaf Ludwigia, and some rather-expensive 'Red' Ludwigia that have never been exposed to the _Myco_ that I can grow out for when I get to the point of restarting my tanks. It's only been set up for about a week, so it's still too early to tell if my hybrid-lighting plan will work. I'd eventually like to shed the artificial light altogether, but I'd like to get it established before I try that.

-I've purchased a larger UV sterilizer of Craigslist to use on the 20gal Permanent QT (the 18w Turbo Twist model), but it's coming from a salt tank and has extensive mineral deposits that I need to clean before I can put it into service.

-I'm not going to do any teardown of the main 52gal until I get the 20gal Permanent QT up and running, so I've done nothing with that so far. The fish inside it are all healthy with extremely strong coloration, so I don't think any of them have active _Myco_ infections at the moment.

Sorry this got so lengthy! I realized it'd been a long time since I did a larger-scale strategy update, so I wanted to be thorough (especially for those who are in this same situation). As always, I welcome any comments/questions/advice you have!


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## KH2PO4 (Jul 18, 2009)

Sterilizing temperature for fish TB.

http://www.who.int/water_sanitation_health/emerging/en/patmycrobact11.pdf


> The resistance of mycobacteria to heat and freezing may influence its survival in
> water systems and in treatment residuals. Certain thermophilic species
> (M. chelonae, M. avium or M. xenopi) survive at temperatures above 55 oC;
> whereas, under the same conditions, M. kansasii or M. marinum are quickly
> destroyed (Merkal & Crawford 1979; Schulze-Röbbecke & Buchholtz 1992).


So heating up the tank to 55 degree celsius (131F) for a few hours might help.

But since there are other species other than M. marinum that cause fith TB. 
76 degree celsius is preferred.

http://www.cfsph.iastate.edu/Factsheets/pdfs/mycobacterium_marinum.pdf


> If trash fish or dead fish carcasses are used as a source of protein
> in the feed for fish, it should be heated at 76oC for 30 minutes to
> kill any pathogenic mycobacteria.


But I don't know how high the temperature our tanks could handle...


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## KH2PO4 (Jul 18, 2009)

KH2PO4 said:


> But I don't know how high the temperature our tanks could handle...


The main interest is probably silicone, right?

Well, I find they are OK with very high temp (like 233 degree celsius).
http://www.americansealantsinc.com/asi-aquarium-sealant/

And the "popular" choice like RTV100 series has continuous operating 
temperature at 200 degree celsius.
http://www.polymax.co.uk/DataSheets/Adhesives/RTV100_Series.pdf

Hmmm. So it should not be a problem to slowly rise the temperature of 
tank's water to 76c ?


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## Diana (Jan 14, 2010)

FlyingShawn,
I have been following this thread, and I commend you on continuing to post your experiences. It must be really hard to have to go through this. Tearing down the whole system, and the worry about the other tanks. 
I appreciate all your research (and the input from others who have posted).

I have a few suggestions about re-stocking. 
Sometimes there are aquarium clubs nearby where you might be able to get plants free or cheaper than buying and paying shipping. 
I do not know if MB lives inside the plant tissues. Perhaps it does live on the outside, in the biofilm that is on the leaves. 
If you get started with some new, young, small plants and grow them out, take cuttings and keep only the cuttings for a few times and keep on growing them in a fish-free environment this might be a way to get plants that are as MB free as possible. 
Plants do not stand up well to sanitizing. But the quick dip in chlorine that removes the biofilm might be all that they need. With no fish in the tank to host the MB, and repeatedly taking only the newest growth and growing it on the plants probably grow faster than the MB spreads. 

Another thought...
Several years ago I bought a product that was supposed to be a collection of beneficial bacteria (not N-cycle bacteria) that would grow in the tank and establish the colonies faster than any 'bad' bacteria. I do not know if this (or a similar product) is still available, but it might be worth looking into. 
The concept is valid: Establish the 'good guys' and maintain conditions for these organisms to thrive, and the 'bad guys' tend to have very low, usually non-threatening populations. Unfortunately UV sterilizer will kill the 'good guys' as well as the 'bad', so you would need to turn off the UV until the colony was established. Still, I think this is a better approach than trying to keep the new set up as sterile as possible.


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## FlyingShawn (Mar 4, 2011)

KH2PO4 said:


> The main interest is probably silicone, right?
> 
> Well, I find they are OK with very high temp (like 233 degree celsius).
> http://www.americansealantsinc.com/asi-aquarium-sealant/
> ...


An intriguing idea, to say the least! Also, excellent find on that WHO article; I'd have never thought to look for a source like that.

The main question on my mind: how? While the tank and silicone may be fine at that temp, I can't for the life of me come up with any ideas for how to safely get the water to that temperature and maintain it there.

The other thing that gives me pause is the spore question. Some of my sources seemed to indicate that _Myco{/I] could enter a spore-stage that would be resistant to those temperatures, whereas the WHO article (at least the parts I've skimmed so far) made no mention of it.

At the very least, this sort of information will greatly increase my confidence in the viability of using my oven to sterilize rocks and substrate. Thanks again!_


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## FlyingShawn (Mar 4, 2011)

Diana said:


> FlyingShawn,
> I have been following this thread, and I commend you on continuing to post your experiences. It must be really hard to have to go through this. Tearing down the whole system, and the worry about the other tanks.
> I appreciate all your research (and the input from others who have posted).


Thanks for the encouragement! And, I can't thank the rest of you who have responded on these threads enough; your input has been totally invaluable as I've tried to wrap my head around these problems!



Diana said:


> I have a few suggestions about re-stocking.
> Sometimes there are aquarium clubs nearby where you might be able to get plants free or cheaper than buying and paying shipping.
> I do not know if MB lives inside the plant tissues. Perhaps it does live on the outside, in the biofilm that is on the leaves.
> If you get started with some new, young, small plants and grow them out, take cuttings and keep only the cuttings for a few times and keep on growing them in a fish-free environment this might be a way to get plants that are as MB free as possible.
> Plants do not stand up well to sanitizing. But the quick dip in chlorine that removes the biofilm might be all that they need. With no fish in the tank to host the MB, and repeatedly taking only the newest growth and growing it on the plants probably grow faster than the MB spreads.


Excellent idea regarding using the cuttings! At this point, I'm not sure if I'm willing to accept any cross-contamination risk through the plants, but I think your idea has enough potential that I need to seriously consider it. I'm especially thinking about if I could adapt that idea to a two-stage emersed-growth idea...

Perhaps I could set up a small, "dirty" (no pun intended) emersed rig outside of my main one and plant cuttings from "contaminated" plants in it (post bleach-dip). After giving them time to grow, I'd make new cuttings from the elevated (airborne) portions of new growth (so no physical contact with the now "contaminated" dirt or water) and transfer them to my "clean" emersed rig to grow out again. The core assumption is that any _Myco_ piggy-backing on the original plant would only spread into areas with direct water contact (ie, the dirt) and not onto the "dry" growth sticking up into the air. Thoughts?



Diana said:


> Another thought...
> Several years ago I bought a product that was supposed to be a collection of beneficial bacteria (not N-cycle bacteria) that would grow in the tank and establish the colonies faster than any 'bad' bacteria. I do not know if this (or a similar product) is still available, but it might be worth looking into.
> The concept is valid: Establish the 'good guys' and maintain conditions for these organisms to thrive, and the 'bad guys' tend to have very low, usually non-threatening populations. Unfortunately UV sterilizer will kill the 'good guys' as well as the 'bad', so you would need to turn off the UV until the colony was established. Still, I think this is a better approach than trying to keep the new set up as sterile as possible.


Do you by any chance recall what that product was called or how it was marketed (ie, what "purpose" it had)? As best as I can recall, I've only ever seen probiotic products related to the N-Cycle. Then again, I can't say that I've ever really looked for them either, so I'll start keeping my eyes out.

My plan for the Permanent QT was to do a complete fishless/plantless-cycle to build up the good bacteria before transferring any "dirty" material over. That way I'll be able to activate the UV unit as soon as I move the fish and plants over and it should have no adverse effect on the already-established good bacteria in the tank. What do you think?


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## Diana (Jan 14, 2010)

I will see if I can dig up any info on the non-N-cycle bacteria. 

Yes, it sounds good to do a fishless cycle, using ammonia, not even fish food or dead shrimp as ammonia source. Then planting, and turning on the UV when the N-cycle is complete. 

I like the idea of growing some plants to the emersed stage, then taking cuttings only of the parts that had no direct contact with the water or soil! Even if MB could hang out in plants somewhat, I would be surprised if it was very mobile in plants. Then a quick dip in chlorine or alcohol, then grow out the cuttings...


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## Diana (Jan 14, 2010)

Here is a pretty long article about using pro-biotics to reduce fish disease. The main focus is on farmed fish, since that is where the money is. However, the concept is valid on a small scale, too (aquariums), the question is where to find the small amount needed. 

http://mmbr.asm.org/content/64/4/655.full

later...

OK. here is an example of this sort of product.

http://www.aquameds.com/medications/aqua-medzyme-dry-concentrate/faq/


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## Diana (Jan 14, 2010)

I am re-reading this thread, and looking into the links more closely. 
In post 85 the link (about disinfectants) specifies that glutaraldehyde is very effective (almost at the end of the article). 
Glutaraldehyde is the liquid carbon source used as a substitute for Excel. 
In low doses it is not toxic to fish. 

If you can get gluteraldahyde, treat the tank and everything else with that, then rinse well. Any trace amount left after rinsing is so low it will not be toxic. 
MAYBE the plants can tolerate the higher dose, too???


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## FlyingShawn (Mar 4, 2011)

Diana said:


> Here is a pretty long article about using pro-biotics to reduce fish disease. The main focus is on farmed fish, since that is where the money is. However, the concept is valid on a small scale, too (aquariums), the question is where to find the small amount needed.
> 
> http://mmbr.asm.org/content/64/4/655.full
> 
> ...


Thanks for finding those articles!

After reading through those, I've spent the last couple hours looking for probiotic supplements along those lines. There are a lot of "beneficial bacteria" products out there, but it seems the vast majority of them are either focused on N-Cycle bacteria or on breaking down waste/mulm in the tank ("Liquid Gravel Vac", etc). However, I have found three that seem like they might be what I'm looking for:

The first is the liquid form of the Aqua MedZyme in your second link:
Aqua MedZyme Liquid
At ~$25 for a 16oz bottle, it sounds pricey, but considering it's supposedly enough to treat 48,000 gallons, I suspect it'll expire long before I run out of it. It's primarily designed for Koi environments, but I'm not aware of any reason why it wouldn't also work for a tropical freshwater tank.

The second is made by the "Dr Tim's" brand, which I believe is fairly reputable for their N-Cycle bacterial products (haven't tried them myself, but I seem to recall seeing that name in a positive light before):
Dr Tim's Aquatics Eco-Balance
A ~$14, 16oz bottle is "only" rated for 480 gallons (quite a difference from the stuff above), but it is specifically marketed for freshwater systems. However, I couldn't find any information the actual bacteria it contains or how many different types there are (according to your links, it's important to have a mix), so I don't know as much about it as I do the first stuff.

The third is this:
Tropical Science Immune-Plus
I've never heard of the brand and it gives no information regarding shelf life, so I suspect that it might not be as high-quality a product as the others. However, it does claim to have 5 different species of bacteria and is supposedly enough for 1700 gallons, so I figured I'd mention it even though I suspect the other two are better options.

I still think the "probiotic" idea is a brilliant strategy going forward (for all of my tanks, not just the quarantine), so I'm definitely going to pursue it. I just need to decide on which product to use for it. Thanks again!


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## FlyingShawn (Mar 4, 2011)

Diana said:


> I am re-reading this thread, and looking into the links more closely.
> In post 85 the link (about disinfectants) specifies that glutaraldehyde is very effective (almost at the end of the article).
> Glutaraldehyde is the liquid carbon source used as a substitute for Excel.
> In low doses it is not toxic to fish.
> ...


Wow, I'm floored by your dedication in looking through all of this again!

OverStocked suggested a Glutaraldehyde-based product in the "Heat Kill"-spinoff thread (Anyone with biology knowledge: boiling or baking to kill Mycobacteria/Fish TB?). While I have no doubt that it's very effective, it's also quite pricey and seems to require some _very_ careful handling, so I felt it was probably overkill for my needs.

To jump back to what you said a couple posts earlier about the "airborne growth" plant strategy:
After some further pondering, I had the thought that even if some _Myco_ were to "climb" up on to the dry growth I want to save, it probably wouldn't find any biofilm to hide in. If that's the case, then that second bleach dip you suggested would probably be enough to clean it off once and for all before transferring the plant to the "clean" emersed rig. Even if some were to manage to hang on, I have to assume that it'd be so little as to fall below "ambient" levels (since all of our tanks supposedly have it) and should be no match for a heavily probiotic-saturated tank environment going forward.

All that being said, I'll probably just use that airborne growth strategy on a handful of my harder-to-find plants (like the Willow Leaf Hygro), since I don't have nearly enough room or patience to grow enough of the other plants to fill-out the main tank when the time comes to start it up again.. For the QT, I'll probably just bleach-dip the plants I want to save and put them in the cycled QT without making any further effort to clean them. After that, it'll be up to the UV and probiotics to keep any _Myco_ at bay.


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## Diana (Jan 14, 2010)

I would tend to trust the Dr. Tim brand. He is the person who isolated the correct nitrogen cycle bacteria when Marineland started marketing it as Bio Spira. 

I cannot be sure if he has gotten on the bandwagon of selling stuff just 'cause it sells or if he is really developing and selling viable options for the aquariums. 
Perhaps a carefully worded e mail to him, including a brief history (like I saw you did for the Seachem question) might help. Here is a link to Dr. Tim's web site:

http://www.drtimsaquatics.com/H2O_PURE_Products/H2O_PURE_Products.html#FW_EB

That Tropical Science label sure looks familiar, but it has been a long time since I used it. Here is a link to their site. I am less inclined to trust this one, because I looked at their cycling product (Nitromax) and it has the wrong bacteria. It has been over a decade that the right bacteria were identified. Any company still selling the old strains is not one I would trust. 

http://www.tsbiolabs.com/product_p/ip-60.htm


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## FlyingShawn (Mar 4, 2011)

Diana said:


> I would tend to trust the Dr. Tim brand. He is the person who isolated the correct nitrogen cycle bacteria when Marineland started marketing it as Bio Spira.
> 
> I cannot be sure if he has gotten on the bandwagon of selling stuff just 'cause it sells or if he is really developing and selling viable options for the aquariums.
> Perhaps a carefully worded e mail to him, including a brief history (like I saw you did for the Seachem question) might help. Here is a link to Dr. Tim's web site:
> ...


I've emailed both Dr Tim's and AquaMeds with specific questions about their products, so we'll just have to see what they say. I suspect that the AquaMeds is a better product overall (just based on the amount of information and research they claim), so my main question for them was if there's any reason it can't be used on a warm-water tropical tank instead of a Koi pond.

Seeing the videos on the Dr. Tim's site about how major institutions (like the Georgia Aquarium) use their products certainly lends them a great deal of credibility in my mind. Like you said, that may not extend to other product lines, but you'd think they'd have major incentive not to risk that reputation.

I think you're right, if Tropical Science is still using the "old" bacteria for the N-Cycle products, that's probably enough reason to doubt their credibility overall. Especially considering the credibility of these other two options, I think it's safe to dismiss them from my consideration.


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## unissuh (Jun 5, 2006)

Might just add a comment while you're on the topic of nitrifying bacteria - a recent paper suggests that the first step of nitrification may not be bacterially driven at all. Rather, the major ammonia oxidizers could actually be archaea. Expect bacterial products to change accordingly within the next few years I suppose...

http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023281

I'm not sure I'd be looking at nitrification cocktails though - looking at heterotrophic bacteria seems like a better idea. These would be more likely to directly compete with (heterotrophic) Myco than true autotrophic bacteria.


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## FlyingShawn (Mar 4, 2011)

unissuh said:


> Might just add a comment while you're on the topic of nitrifying bacteria - a recent paper suggests that the first step of nitrification may not be bacterially driven at all. Rather, the major ammonia oxidizers could actually be archaea. Expect bacterial products to change accordingly within the next few years I suppose...
> 
> http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023281


Very interesting, thanks for summarizing it for us! (I make no secret that I'm not a biologist, so I would have struggled to understand the article without going into it with the "gist" as you explained it)



unissuh said:


> I'm not sure I'd be looking at nitrification cocktails though - looking at heterotrophic bacteria seems like a better idea. These would be more likely to directly compete with (heterotrophic) Myco than true autotrophic bacteria.


Just a point of clarification: we're not looking at nitrification cocktails in this discussion. Rather, we were using our limited knowledge of that field (that Dr. Tim's research identified the correct bacteria involved in the N-Cycle and Tropical Science are still marketing the incorrect/less-correct ones from the prior understanding) as a ruler by which to judge those companies' scientific credibility when comparing their non-N-Cycle "probiotic" products.

Granted, we still don't know much about what kind of probiotic bacteria these companies are marketing (aside from AquaMed, who claims it's a "specilized bacillus bacteria"), but we aren't specifically looking at nitrification products either.


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## unissuh (Jun 5, 2006)

Apologies - I obviously got the wrong impression (that you were looking for nitrification products)! I'm not going to be much help on products that are available, but your approach in picking reputable companies sounds perfectly reasonable for me. If you were in AU, I'd recommend Aquasonic, a local company which makes high class products but strangely has a very low key reputation amongst hobbyists.

An additional small suggestion for when you start up the next tank - if you can find someone who has a well established tank already running, get 'em to do a good gravel vacc and dump all the mulm in your (clean) substrate. I think you will find that you can transfer a heck of a lot of beneficial microflora this way. May help if you have them all in there from the get go.


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## FlyingShawn (Mar 4, 2011)

Just heard back from Dr. Tim's.

Following a brief explanation of my situation, I asked them three specific questions about their product:

1) Does Eco-Balance contain multiple strains of bacteria? If so, how many?
2) After several months of dosing, are the beneficial bacteria in this product able to form a stable self-sustaining population or does the tank require regular dosing to sustain these populations?
3) Has any _Mycobacterium_-specific research been done to see if these probiotic bacteria have a competitive advantage against it when competing for resources?

Here is their response:


> Hello Mr __________:
> 
> Thanks for your interest in our products.
> 
> ...


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## mjbubbles (Aug 26, 2011)

FS, thanks so much for your dedication in sharing all this with us. I really appreciate it. I'm sorry you've had to go through the experience, but your sharing will help others who find them selves in the same boat.

Having just euth'd a white cloud mtn minnow with what appears to be mycobacterium, I am concerned about the other fish. I'll be keeping an eye on them and returning to this thread to check on your progress as well. I hope the probiotics and all your other approaches help and you have no more sick fish.

Take care,
mj


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## FlyingShawn (Mar 4, 2011)

I haven't abandoned you! I'm really sorry that I haven't posted any updates lately, I've been really busy at work and what little time I've had at home has been occupied with a couple major projects (like this one!) and trying to relax a little every once in a while..

There hasn't been much to report either: I've been working on sterilizing a lot of various pieces of equipment and decorations, building my "Clean" and "Dirty" emersed rigs, and setting up the 20gal permanent QT we've spent so much time discussing.

I'll try to make up for the delay with a few pictures of what I've been doing...

First off, I set up my "Clean" emersed rig. This one will only be used for plants that have never been exposed to the TB (at this point that list is limited to the plants that were in my jury-rigged and soon-to-be-decommissioned setup for growing HC emersed before setting up the big tank last time around). Eventually, I'll also move over the "deemed-clean" cuttings from the "Dirty" emersed rig, like Diana and I were talking about earlier.

It's made using a Sterilite tub, cut-up sections of water bottles, and a mix of artificial and natural lighting. I punctured small holes in the bottom of each bottle so that the water could get in and lined one wall of the rig with aluminum foil to try to capture as much natural sunlight from the window as possible. I don't know if the foil is making any real difference to speak of, but what can it hurt to try? Originally, I used the body from a Finnex 26w clip light for the artificial side of the lighting, but between a couple weeks of a lot of clouds and the weakness of the light getting through the lid, the plants were hanging on with little-to-no growth to speak of.



So, I decided to build a new lighting system that would work better for this rig. I purchased a cheap incandescent hood for a 10gal tank and a couple of Walmart fluorescent aquarium lights, mounted the fixture from the hood to the inside of the lid, and added a couple of combination reflectors/bulb protectors made out of water bottles and aluminum foil. Here's how it turned out:




Sorry, I don't have any recent pictures of the interior of the rig, but after a couple of weeks of mostly-sunny days and the new lighting, the growth inside has absolutely exploded! I've used these same bulbs before in a Walstad-style 10gal tank and was also really impressed with how well the plants did under them in there.

Speaking of that 10gal and those other bulbs, I also just finished the "Dirty" emersed rig for the plants I want to preserve from the contaminated tanks. I've cut the bottles a little taller than normal and made clippings of the plants so that they (mostly) fit within the height of each bottle. Assuming they successfully transition to emersed-mode and grow well, I'll make cuttings from the parts that grow out the top of each bottle and move them into the Clean emersed rig (after a quick bleach dip, to be safe). Here's how it looked after I set it up a few days ago:



I'm mainly using it to grow the Willow Leaf Hygrophilia that I like so much (and haven't seen elsewhere), but I had extra room in the tank and decided to toss a couple other plants in there just to see how they'd do emersed.

The last thing to update you on is that I've just begun cycling the 20gal permanent QT tank. I set it up on Friday, added a little ammonia, and then poured in Tetra SafeStart on Saturday afternoon. I've tried using SafeStart before without a huge amount of success, but after reading through this Q&A with Tetra I figured out that my water conditioner was likely also killing off the SafeStart bacteria by detoxing the ammonia solution it comes in:
SafeStart Questions Answered

So far the cycle is off to a very promising start. When last I measured it, it'd converted 1ppm of ammonia into nitrate within the span of 6 hours! My plan is to keep dosing ammonia until the tank is very-heavily cycled, put in plants, and then move the fish over from the big tank. Then I'll be able to start tearing down/sterilizing the 52gal itself.

The teardown of the 52gal will be a hugely time-consuming project and I have vacation later this month (which I'm hoping to get out of town for), so updates will continue to be infrequent for the time being. In the meantime, I continue to welcome any ideas or suggestions you guys may have!


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## FlyingShawn (Mar 4, 2011)

mjbubbles said:


> Having just euth'd a white cloud mtn minnow with what appears to be mycobacterium, I am concerned about the other fish. I'll be keeping an eye on them and returning to this thread to check on your progress as well.


Good luck MJ, I really hope that your minnow had something(anything!) else! 

I think I've concluded that the 2nd worst thing about _Myco_ is that it's so hard to diagnose and, once you learn it exists, you're always paranoid that your fish may have it (the worst thing of course being that it can also infect _you_!).

I forgot to add earlier that the all remaining fish still look healthy. As before, there's one female Odessa Barb that I keep thinking might be in the early stages, but she doesn't seem to be getting any worse and I think my mind is just playing tricks on me.

Never heard back from AquaMed, so I've decided to go ahead and purchase the Dr. Tim's probiotic product and will start dosing that on the permanent QT once it finishes cycling.


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## wkndracer (Mar 14, 2009)

Thought I would go ahead and add some current information and links I've gathered into this thread rather than have another floating on the forum. I won't call it "fish TB" though. 

The person leading me through the information has told me that's yesterday's best guess and not a correct term to use.
Links with correct information:
https://srac.tamu.edu/index.cfm/event/getFactSheet/whichfactsheet/231/
https://srac.tamu.edu/index.cfm/event/getFactSheet/whichfactsheet/235/
https://srac.tamu.edu/index.cfm/event/getFactSheet/whichfactsheet/238/
http://sfrc.ufl.edu/fish/outreach/extpubs/
http://www.peter.unmack.net/archive/rml/rmlaug99/0117.html
http://z7.invisionfree.com/MTSA/ar/t24.htm
http://www.tnfish.org/FishDiseasesParasites_TWRA/files/Mycobacteriosis.pdf

Information can be found under the tab “Health Services” on this site.
There’s a lot of good information there including how to produce clean fish. 
(Unfortunately they also call myco fish T.B. in one or two places.) 
http://zebrafish.org/zirc/home/guide.php
http://zebrafish.org/zirc/health/diseaseManual.php#Mycobacteriosis (Fish TB)

within this reply I've provide all the best primary information reference I have been given that’s not copy write restricted.


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## wkndracer (Mar 14, 2009)

I just finished going through this thread again and all related links today.
Some of the research papers I posted in my post above duplicate some you had already found.

On the bleach testing mentioned it was brand specific.
Cleaning methods with bleach at normally considered ratios won't kill all myco strains. The 10% once reported effective is now considered wrong. Tested bleach washes strong enough to kill myco was also strong enough to damage tank sealants. *Clorox Ultra* was tested as only 'moderately' effective (50,000mg/L) with >20 minutes nessary to eliminate growth. But bleach is still part of it. Bleach will clear the biofilm to help expose the bacteria. H2O2 above 3% can also be effective for that part. But it's the wash of 50-70% alcohol that Doc say's kill it. Alcohol was effective within 1 minute so a spray bottle will work for surface cleaning rather than a soak. (Journal of Aquatic Animal Health 2005)

Glutaraldehyde activates effectively in alkaline solutions. Effective on myco at 2% and above I believe (but need to verify by the paper I have at home). Excel strength is reported to test out as 1.5% in two posted lab tests I've been able to find. 

Wet heat will work according to the Doc I'm following advice from.
Thinking turkey fryer and 30 minute boil to raise large wood cores, gravels, Flourite etc. completely to temperature. Even new soils will be boiled or baked here.

For dry heat 350°F is what was approved in query. 

3-5 individuals from all fish orders received going forward will be sacrificed in the name of safety before the rest leave my entry quarantine tanks.

Yeah you guess correctly,,, a 'friend' brought myco into my entry tanks and I have been evaluating cross contamination potentials for the last month. 

LMAO! I just realized this is my first forum post declaring I have myco positive fish. :icon_roll Those that have PM'ed for plants and fish and haven't received any answer now know why. I won't share crap like this. Like Shawn I'm figuring how far it has gotten and I slammed the door shut on the mailman in December. 
Those that received critters prior to that from me they were clear of it and I'm certain of that.


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## Diana (Jan 14, 2010)

Thank you wkndracer for this update. 
Now back to read those links!


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## wkndracer (Mar 14, 2009)

Shawn and I exchanged posts in another thread on this topic recently and using heat to kill myco (the temperature needed) was one of the things briefly talked about. Locating open source (allowed for copy or link) isn't easy.

Here's one open source lab study protocol excerpt quoted here with the ref. link. 
This is regarding water boiling (212°F) Deactivating = kill. 
I have compiled a number of PDF documents and web links to lab and medical study regarding viable temperature range for a large number of myco strains. Reading several of them at once will send my brain into a spin LOL

_*"For procedures that do not require intact, high quality DNA such as PCR testing, our laboratory depends on the much more expedient lysate method of boiling culture at 100°C for 10 minutes, followed by mechanical lysis for 2 minutes to release DNA. Although crude, this procedure is adequate for our PCR testing needs, has been shown to completely destroy live organism in our laboratory, and is consistent with other studies [7,8]. The study by Zwadyk et al. concluded that inactivating mycobacteria by heat lysing at a temperature of 100°C for 30 minutes did not inhibit its ability to be amplified by PCR or strand displacement amplification [8]. Furthermore, this study has shown that inoculating the boiled lysate alone without mechanical lysis was adequate in rendering the sample non-viable.

The viability testing of the two methods outlined above for DNA extractions were performed by various technicians who routinely follow these procedures, lending interpersonal variability to the study. It was demonstrated that the small nuances to procedure, such as the varying density of culture used, did not affect the method employed to deactivate the organism."

*_Research article tittle:
Viability testing of material derived from _Mycobacterium tuberculosis _prior to removal from a Containment Level-III Laboratory as part of a Laboratory Risk Assessment Program.
http://www.biomedcentral.com/1471-2334/5/4


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## FlyingShawn (Mar 4, 2011)

Wnkracer, what are the latest thoughts on the idea of using "sentinel" fish as part of a quarantine protocol to help screen new arrivals for _Myco_?

If memory serves, I first saw the idea proposed by Walstad in one of the articles I read during my initial research (although I'm afraid I've forgotten which one and am not even positive it was Walstad). The idea was to keep a population of Zebra Danios (which I seem to recall are especially susceptible to _Myco_, potentially even more so than Rainbowfish) in quarantine for an extended period of time (6+ months) to ensure they are healthy, move them into a non-quarantine system like a display tank, and then move a couple of them back into your quarantine tank each time you have a new arrival.

You'd still have to keep the new arrival in quarantine with the Zebras for a long time (I'd guess a minimum of 45-60 days), but their higher susceptibility would theoretically allow you to be more confident of the new arrival's health at the end of that time.

My main tank is still not up and running again and will likely be a while (I'm hoping to get a more detailed update posted before too much longer), so I was thinking that now might be a good time to get the Zebras and put them in the initial quarantine if that's still considered a helpful precaution.

Have you seen anything in the more recent literature that references this concept?


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