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Thread: UPDATE: IT'S FISH TB. ...Help me diagnose this disease that's killing my fish Reply to Thread
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Topic Review (Newest First)
07-14-2013 09:53 PM
FlyingShawn 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?
07-03-2013 04:04 PM
wkndracer 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
01-26-2013 03:29 AM
Diana Thank you wkndracer for this update.
Now back to read those links!
01-25-2013 08:57 PM
wkndracer 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. 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.
01-22-2013 10:19 PM
wkndracer 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/even...factsheet/231/
https://srac.tamu.edu/index.cfm/even...factsheet/235/
https://srac.tamu.edu/index.cfm/even...factsheet/238/
http://sfrc.ufl.edu/fish/outreach/extpubs/
http://www.peter.unmack.net/archive/...ug99/0117.html
http://z7.invisionfree.com/MTSA/ar/t24.htm
http://www.tnfish.org/FishDiseasesPa...acteriosis.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/dis...s%20(Fish%20TB)

within this reply I've provide all the best primary information reference I have been given that’s not copy write restricted.
04-03-2012 12:55 AM
FlyingShawn
Quote:
Originally Posted by mjbubbles View Post
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.
04-03-2012 12:45 AM
FlyingShawn 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!
03-17-2012 06:31 AM
mjbubbles 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
03-06-2012 08:00 PM
FlyingShawn 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:
Quote:
Hello Mr __________:

Thanks for your interest in our products.

Our probiotic for freshwater has 6 species for saltwater/reef it has 8 species.

Our probiotic strains were isolated by us from aquaria and are grown by us - almost no other company in the Aquatics Industry can claim this.

The beneficial bacteria do not form self-sustaining populations because they are eaten by protozoa and other micro-organisms so regular dosing is necessary.

Regarding mycobacterium specifically - at this point there is no published research but hopefully before the end of the year there will be a paper or two.

Hope this answers your questions but always available should you have more

Cheers
DrTim's Aquatics
03-06-2012 03:02 AM
unissuh 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.
03-06-2012 02:01 AM
FlyingShawn
Quote:
Originally Posted by unissuh View Post
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:...l.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)

Quote:
Originally Posted by unissuh View Post
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.
03-06-2012 01:41 AM
unissuh 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:...l.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.
03-06-2012 12:11 AM
FlyingShawn
Quote:
Originally Posted by Diana View Post
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_PU...cts.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
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.
03-04-2012 06:16 PM
Diana 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_PU...cts.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
03-04-2012 02:20 AM
FlyingShawn
Quote:
Originally Posted by Diana View Post
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???
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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