# Anyone with biology knowledge: boiling or baking to kill Mycobacteria/Fish TB?



## OverStocked (May 26, 2007)

The decorations/lava rock/wood will be fine with the chemicals. I highly recommend Metricide 14, as well, which is used as an excel replacement by many here. It is a surgical disinfectant 

Metricide 14 has a TB kill time of 3 minutes. Any parts you're not attached to should be disposed of. Substrate, for instance.


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## FlyingShawn (Mar 4, 2011)

OverStocked said:


> The decorations/lava rock/wood will be fine with the chemicals. I highly recommend Metricide 14, as well, which is used as an excel replacement by many here. It is a surgical disinfectant
> 
> Metricide 14 has a TB kill time of 3 minutes. Any parts you're not attached to should be disposed of. Substrate, for instance.


Wow, that stuff is PRICEY!! While the 3 minute TB kill time is certainly attractive, I simply can't justify $28/gal when I have this much to clean (52gal main tank, plus multiple smaller tanks)

Regardless of which chemical I use, I'm still concerned about it soaking into driftwood or the lava rocks and either taking excessively long to soak back out or potentially being toxic to the tank.

The substrate is MTS capped with Flourite. Seachem actually recommends boiling Flourite as a way of sterilizing it. So, if I can confirm that heat-killing will work, doing so would save me a _substantial_ amount on replacing enough Flourite for a 52gal tank!


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## OverStocked (May 26, 2007)

In a spray bottle that 1 gallon woudl last you all day. Look it up on dealmed. Boiling sanitizing, but does not sterilize. It requires an autoclave to sterilize.


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## FlyingShawn (Mar 4, 2011)

OverStocked said:


> In a spray bottle that 1 gallon woudl last you all day. Look it up on dealmed. Boiling sanitizing, but does not sterilize. It requires an autoclave to sterilize.


For cleaning the tanks themselves, a spray bottle would work. However, I wouldn't trust a spray-application for getting into all of the little pores of lava rock or the driftwood. Regardless, I'd still be concerned about toxic chemicals remaining in the rocks and driftwood after I was done.

After doing a little reading, it seems that dry heat also works for sterilizing, which takes me back to the other option I mentioned of using the oven to bake these materials.


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## FlyingShawn (Mar 4, 2011)

unissuh said this in the original thread:


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: as far as he (and I) know, _marinum_ is a low-temp bug. At least on my part, that's only a semi-educated guess, so I'm really hoping to find someone on here who can confirm that with some confidence/authority.

Please chime in if you have any of the education/knowledge to help me figure this out!


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## mistergreen (Dec 9, 2006)

http://en.wikipedia.org/wiki/Mycobacterium_marinum



> The inhibition of growth of M. marinum at 37°C


I'm pretty sure 100°C will kill it.


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## Sindawe (Oct 5, 2008)

mistergreen said:


> http://en.wikipedia.org/wiki/Mycobacterium_marinum
> 
> 
> 
> I'm pretty sure 100°C will kill it.


A quick search turns up this article that indicates _Mycobacterium marinum_ is a spore forming organism.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2705590/

Its been awhile since I worked in the Biosciences, but IIRC you're going to need to autoclave soft materials like drift wood at 121 Celcius, 15 PSI pressure for not less than 20 minutes to get effective sterilzation of spores.


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## mistergreen (Dec 9, 2006)

We're not going to find an autoclave cheaply. Try a pressure cooker.


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## mistergreen (Dec 9, 2006)

FlyingShawn said:


> Regardless of which chemical I use, I'm still concerned about it soaking into driftwood or the lava rocks and either taking excessively long to soak back out or potentially being toxic to the tank.


It's also toxic to you. You need training to handle the stuff.


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## Chlorophile (Aug 2, 2011)

mistergreen said:


> It's also toxic to you. You need training to handle the stuff.


And let me tell you those training videos are awful.
I had to get my pesticide handler/worker training a few weeks ago and then I decided never to handle pesticides because they made it clear that you're going to get sick and get cancer and get nerve damage =[


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## kwheeler91 (May 26, 2009)

I would ditch all things except the tank and filter housings, sterilize tank and filters, replace media and everything else including substrate, wood, plants, fish. I know its a pain but I have heard fish tb can be contracted by humans. Even if thats not the case you dont want a reoccurence once you get set up again. There are many generous members here who might be able help you get back on your feet.


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## DogFish (Jul 16, 2011)

OverStocked said:


> ....Any parts you're not attached to should be disposed of. Substrate, for instance.


roud:

I'd bleach out the tank & filter. I can't believe much would survive a boil or a bake at 200 degrees. 

Sometimes practical is the better path than OCD. If you need to spend $30 in meds& chemicals to save a $3 pc of drift wood, the math on that really doesn't make much sense.


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## mallorieGgator (Jan 25, 2012)

Unfortunately, heat as in boiling or baking won't work. You would have to autoclave everything (121.21 degrees celcius) for about 90 minutes to be 100% sure you killed it. If it forms spores, you will have to be careful about even using chemicals as some bacteria's spore can actually resist things such as bleach and alcohol. I know with bacillus bacteria they form spores and contaminate a lot of things in the micro lab I used to work in. If we heated a sample to less than 81 celcius, then bacillus would live in a spore form but everything else that was non-spore forming would die.


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## DogFish (Jul 16, 2011)

mallorieGgator said:


> Unfortunately, heat as in boiling or baking won't work.


This seems to contradict that http://textbookofbacteriology.net/control_3.html

I can understand the consistency & effectiveness of the autoclave. I don't understand without documented study of how to destroy Mycobacterium marinum, how can we say that Boiling at 220 or heat in a oven at 200 is absolutely not going to work?


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## TexasCichlid (Jul 12, 2011)

Obviously you are pretty worried about it. Fish TB is not a joke. Why not have some peace of mind and just get a new setup? Bleach the tank, and start completely over. Your biggest culprits will be the filter materials, hardscape and substrate. With the amount of work and worry you have put into this, your opportunity costs have probably already exceeded actual costs of purchasing the replacement bits you need.


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## mallorieGgator (Jan 25, 2012)

I just know that I work with bacteria daily and the only sure-fire way we destroy it is in an autoclave.


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## FlyingShawn (Mar 4, 2011)

DogFish said:


> This seems to contradict that http://textbookofbacteriology.net/control_3.html
> 
> I can understand the consistency & effectiveness of the autoclave. I don't understand without documented study of how to destroy Mycobacterium marinum, how can we say that Boiling at 220 or heat in a oven at 200 is absolutely not going to work?


Although an autoclave would seem to be the best method, I'm afraid I don't have access to one.

Take a look at the "Dry Heat" section of this article:
Wikipedia - Sterilization (microbiology)

Yes, I know it's Wikipedia, but after doing a LOT of other searching online, the other sources I could find seemed to be consistent with what it said: A "standard" dry heat application is 160C/320F for 2 hours (starting after the item is thoroughly heated), with some sources recommending closer to 6 hours. It also seems that increasing the temp by 10C roughly halves the required time.

To be safe, I'm thinking of more like 400-450 F for a couple hours. For the substrate, I'd only do 25% at a time to make sure it's thoroughly heated in a reasonable amount of time.

I've also seen references to an "intermittent boiling" process that's designed to kill spores by boiling for 30 minute increments spaced 24 hours apart for several days. The idea is that the end of each heat cycle encourages the bacteria to come out of their spore-stage, so the next heat cycle kills them. I'm thinking of using this method for a couple of irreplaceable decorations that I'm not sure would be safe at the temperatures used in the dry heat method above.


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## FlyingShawn (Mar 4, 2011)

Chlorophile said:


> And let me tell you those training videos are awful.
> I had to get my pesticide handler/worker training a few weeks ago and then I decided never to handle pesticides because they made it clear that you're going to get sick and get cancer and get nerve damage =[


I've seen enough training videos to know how painful they are, and these sound _much_ worse than the ones I've been subjected to!


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## FlyingShawn (Mar 4, 2011)

TexasCichlid said:


> Obviously you are pretty worried about it. Fish TB is not a joke. Why not have some peace of mind and just get a new setup? Bleach the tank, and start completely over. Your biggest culprits will be the filter materials, hardscape and substrate. With the amount of work and worry you have put into this, your opportunity costs have probably already exceeded actual costs of purchasing the replacement bits you need.


Unfortunately, a completely new setup isn't an option due to cost reasons.

By the time you factor in the time/labor to do a full tear-down of the system and purchasing/preparing new components (ex: rinsing Flourite), the actual time difference between a "cleaned" setup and a "new" setup isn't that substantial. The only real way it'd make a difference was to replace the tank itself (rather than the components), but since it's a 52gal flat-back hex and not a standard 55gal, even that doesn't make sense economically.

Even then, I don't make enough for my "lost time" opportunity cost to make the difference (contrary to what most believe, flying isn't very lucrative: airline pilots start out around 20k and it doesn't go up quickly from there)


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## FlyingShawn (Mar 4, 2011)

mallorieGgator said:


> I just know that I work with bacteria daily and the only sure-fire way we destroy it is in an autoclave.


I understand that's what you use and it's probably the "best" method, but do you have any reason to believe that a "dry heat" process liked the one I linked to above wouldn't also be effective? Yes, it'd require more heat and take significantly longer (which is why an autoclave is the preferred method), but I couldn't find anything that would imply that it wouldn't work altogether.


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## mrdavco (Jan 2, 2018)

The following might be helpful. 

Dr Julie Ball of Extreme Sportfishing has some interesting takes on Myco. Due to the fact this is my 1st post, I can not place the Link as part of this post as per Planted Tank only allows posting a Link AFTER you author 5 postings on the forum? 

Just Google "fish handlers disease Dr Julie Ball" and it will take you directly to article. 

I Copied n pasted from Dr Julie Ball's forum on marine injuries and Myco, The Fish Handler's Disease and why Myco marinium is usually only found in the extremities and not the core body. 

Dr Julie's statement: "The most frequently affected areas tend to be the extremities, which include the arms, legs, hands, feet, fingers and toes. This is the case for two reasons: first, because the extremities are the most likely part of the body to be exposed to an injury while handling and maneuvering around infected organisms or objects, and secondly because the Mycobacterium marinum bacteria cannot proliferate in a warm body environment, therefore the cooler temperature of the arms and legs lend to a more suitable home for the unwelcome visitors."

I live in Miami, my family eats raw and roasted oysters, crab stew along w/fried turkey and smoked brisket for Thanksgiving and Christmas. Being and avid fisherman and waterman I deal with Myco a lot- usually in a finger but sometimes the hand. Never had it in my arms or any other places.

To get rid of the infection I make a really hot coffee cup of super-saturated salted water (1/4cup of salt minimum) and then soak my affected finger for about 30-40 minutes, frequently reheating the salt solution in the microwave. I try to get the finger as hot and red as I can stand it, to elevate the finger temperature. This does 2 things: 1. eases the pain and 2. I think, destroys the Myco bacteria. 

After soaking I smear a copious amount of 20% strength of Ichtammol Ointment over the affected area, smear some more ointment on a 4x4" gauze sponge along the edge and place the edge over the area , wrap the sponge around the finger and then tape it firmly- not tight, into place and leave it undisturbed for a day or two. Some people will remember the black and greasy smelling goo as the stuff their mom put on boils to draw them to a head. The ointment seems to draw out the infection and all is well in a couple of days. I do this soaking and dressing at night while watching TV. The next morning, all pain and most of the swelling is gone and a couple days later all is good. I've yet to go to a doctor for treatment. 

You might find this is way out in left field as for your problem but the point is- Myco can't live in body-core temps so if you can get Myco hot, it dies. Normal Core body temp is usually between 96-98F degrees. 

Seems to me if the bacteria can't survive in the core body and usually thrives in the extremities because of the cooler tissue then if you elevate the finger temp higher than core temp, the bacteria should die. All I can say is it works for me. 

I would try to elevate temp of aquarium to 110 degrees or higher and leave it like that for awhile and then do a culture test for Myco. If you are a good handyman you can jury-rig a hot water heater element from HomeDepot and you might have your solution to your problem. Be careful and hang the element in the aquarium all set-up the way it is and just keep it hot for a couple of days. You can then test it by having a lab grow cultures for Myco.


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## Hyue (Oct 11, 2017)

This is a long scholarly article but it describes how a university zebrafish facility dealt with an M. marinum outbreak. They apparently used hypochlorite solutions along with hot water to sterilize their tanks. Look at page 16 onwards. 

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4931754/


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## roadmaster (Nov 5, 2009)

FlyingShawn said:


> The short version of the story is that my fish are confirmed to be infected with _Mycobacterium Marinum_ (aka "Fish TB") and I'm looking at a full-system teardown and nuke. For the tank and filter system, I'll be using a chemical warfare protocol (high-concentration bleach, then Lysol, then 70% isopropyl alcohol), but am afraid a chemical strategy would be damaging to decorations/lava rock/driftwood. Will heat work?
> 
> It seems to be an accepted procedure to sanitize driftwood by boiling or baking it, but I have not been able to find a reliable answer on if it'll work on _Mycobacteria_.
> 
> ...


See Vinegar for killing mycobacterium and cleaning the tank,filter parts.
Get new substrate .
Boil the wood.
My two cent's.


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## roadmaster (Nov 5, 2009)

Would also be of help to other's to know how the determination of TB was arrived at?
Without gill scraping, or necropsy,might it just as easily be one of a few varieties of viruses such as Herpesviruses,Rhabdoviruses,Iridoviruses, or one of a few gram positive/negative bacterium other than TB which is rare, but often misdiagnosed.??
Tried to find thread that OP linked to but draw a blank page.
Tried going back through a few pages of post's in Fish sub forum but no luck.


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## FlyingShawn (Mar 4, 2011)

mrdavco said:


> The following might be helpful.
> 
> Dr Julie Ball of Extreme Sportfishing has some interesting takes on Myco. Due to the fact this is my 1st post, I can not place the Link as part of this post as per Planted Tank only allows posting a Link AFTER you author 5 postings on the forum?
> 
> Just Google "fish handlers disease Dr Julie Ball" and it will take you directly to article.


To save everyone a step, here is the article in question: Dr. Julie Ball - Extreme SportFishing 

It's a worthwhile read, both because it presents another perspective on _Myco_ from outside the bubble of the aquarium community and because it gives you some idea of what a human treatment regimen might look like if you get a _Myco_ infection (something we often gloss over in our focus on treating our tanks). Thanks for sharing it!



mrdavco said:


> To get rid of the infection I make a really hot coffee cup of super-saturated salted water (1/4cup of salt minimum) and then soak my affected finger for about 30-40 minutes, frequently reheating the salt solution in the microwave. I try to get the finger as hot and red as I can stand it, to elevate the finger temperature. This does 2 things: 1. eases the pain and 2. I think, destroys the Myco bacteria.
> 
> After soaking I smear a copious amount of 20% strength of Ichtammol Ointment over the affected area, smear some more ointment on a 4x4" gauze sponge along the edge and place the edge over the area , wrap the sponge around the finger and then tape it firmly- not tight, into place and leave it undisturbed for a day or two. Some people will remember the black and greasy smelling goo as the stuff their mom put on boils to draw them to a head. The ointment seems to draw out the infection and all is well in a couple of days. I do this soaking and dressing at night while watching TV. The next morning, all pain and most of the swelling is gone and a couple days later all is good. I've yet to go to a doctor for treatment.


For the sake of clarity, it should be noted that neither of these treatments (hot salt solution and/or Ichtammol Ointment) are mentioned in the quoted article or any other research literature I've come across. While both of those may help to alleviate your symptoms, the only treatments mentioned or accepted within the medical community seem to be antibiotic-based (unless there's some new research I'm unaware of).



mrdavco said:


> You might find this is way out in left field as for your problem but the point is- Myco can't live in body-core temps so if you can get Myco hot, it dies. Normal Core body temp is usually between 96-98F degrees.
> 
> Seems to me if the bacteria can't survive in the core body and usually thrives in the extremities because of the cooler tissue then if you elevate the finger temp higher than core temp, the bacteria should die. All I can say is it works for me.


Dr. Ball: "...and secondly because the Mycobacterium marinum bacteria cannot proliferate in a warm body environment, therefore the cooler temperature of the arms and legs lend to a more suitable home for the unwelcome visitors."

There is a significant distinction that needs to be made between slowing/halting the proliferation of _Myco_ and actually *killing* it; _Myco_ is an uncommonly hardy pathogen and eradicating it requires minimum temperatures more in the 60C/140F range (see the article Hyue shared a few posts up).


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## FlyingShawn (Mar 4, 2011)

Hyue said:


> This is a long scholarly article but it describes how a university zebrafish facility dealt with an M. marinum outbreak. They apparently used hypochlorite solutions along with hot water to sterilize their tanks. Look at page 16 onwards.
> 
> https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4931754/


Thank you, Hyue, this was a fascinating read! The fact that they had to go to such extreme measures just to *manage* _Myco_ without eradicating it is an excellent example of why hobby aquarists should take possible Myco cases seriously and not share fish/plants/equipment from potentially-infected tanks with other hobbyists! 



roadmaster said:


> See Vinegar for killing mycobacterium and cleaning the tank,filter parts.
> Get new substrate .
> Boil the wood.
> My two cent's.


Vinegar is certainly useful in this hobby, but I am not aware of any research indicating it to be effective against _Myco_, care to share your source?



roadmaster said:


> Would also be of help to other's to know how the determination of TB was arrived at?
> Without gill scraping, or necropsy,might it just as easily be one of a few varieties of viruses such as Herpesviruses,Rhabdoviruses,Iridoviruses, or one of a few gram positive/negative bacterium other than TB which is rare, but often misdiagnosed.??
> Tried to find thread that OP linked to but draw a blank page.
> Tried going back through a few pages of post's in Fish sub forum but no luck.


You'd have to go MUCH farther back to find it; all of the threads related to my TB infection have been dormant for 5-6 years (you can imagine my surprise when I got the notification that I had new replies!)! Here is the original thread that led to the necropsy diagnosis of TB in my fish:
UPDATE: IT'S FISH TB. ...Help me diagnose this disease that's killing my fish


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## JusticeBeaver (Oct 28, 2017)

FlyingShawn said:


> Thank you, Hyue, this was a fascinating read! The fact that they had to go to such extreme measures just to *manage* _Myco_ without eradicating it is an excellent example of why hobby aquarists should take possible Myco cases seriously and not share fish/plants/equipment from potentially-infected tanks with other hobbyists!
> 
> 
> 
> ...


Mycobacteria are a bane to most science labs as well. We frequently test our cell cultures for mycobacteria contamination. What makes it particularly difficult to deal with is that it can replicate within the host cell so it's very hard to treat since you'd need to kill the cell or have a delivery method which can penetrate both the mammalian cell and affect the mycobacteria. Also many mycobacteria are acid-fast so I'm not sure vinegar would be very useful anyway. 

As for sterilization techniques people could use, the simplest one is to get a pressure cooker and boil your stuff that way. It only takes about 15-20 minutes for a good sterilization unless your material is incredibly porous and dense. It's basically a mini autoclave.


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## roadmaster (Nov 5, 2009)

I happened upon the use of vinegar while reading study from American Society for Microbiology some time back.(with regard to iridoviruses)
Sorry,my skills are limited with regard to sharing links but one might google.."Vinegar to kill Mycobacterium"
Tried to link to thread provided, "Full details microscopic slides" but draw blank page.


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## FlyingShawn (Mar 4, 2011)

roadmaster said:


> I happened upon the use of vinegar while reading study from American Society for Microbiology some time back.(with regard to iridoviruses)
> Sorry,my skills are limited with regard to sharing links but one might google.."Vinegar to kill Mycobacterium"
> Tried to link to thread provided, "Full details microscopic slides" but draw blank page.


Here is the research study your recommendation led me to:
mBio: Acetic Acid, the Active Component of Vinegar, Is an Effective Tuberculocidal Disinfectant

It seems acetic acid (in higher concentrations than found in common vinegar) is effective against several species of _Mycobacterium_, but I could not find any research testing it against _M. Marinum_. Maybe it works, maybe not (especially given JusticeBeaver's comments about being acid-fast), but given the severity of the threat _Myco_ poses I would strongly caution against relying on it to treat _M. Marinum_ without more data.

It's strange that the link is broken. Here is a permalink to the post in question, but it seems that imgshack dropped their free tier sometime in the last six years. If I get a chance, I'll try to find a new platform and edit that original post with new picture links (UPDATE: done!).
Post #49

I'd also recommend reading posts #50 (more info about _Myco_ from Dr. Palmeiro) and #61-62 (regarding high salt concentrations and Dr. Palmeiro's opinion that it'd be ineffective against _Myco_).



JusticeBeaver said:


> Mycobacteria are a bane to most science labs as well. We frequently test our cell cultures for mycobacteria contamination. What makes it particularly difficult to deal with is that it can replicate within the host cell so it's very hard to treat since you'd need to kill the cell or have a delivery method which can penetrate both the mammalian cell and affect the mycobacteria. Also many mycobacteria are acid-fast so I'm not sure vinegar would be very useful anyway.
> 
> As for sterilization techniques people could use, the simplest one is to get a pressure cooker and boil your stuff that way. It only takes about 15-20 minutes for a good sterilization unless your material is incredibly porous and dense. It's basically a mini autoclave.


Do you happen to test for _M. Marinum_ specifically or just _Mycobacteria_ in general? If you happen to find an _M. Marinum_-contaminated culture, is there any chance you'd have the opportunity to culture it and try some of the acetic acid protocols from the mBio article above?

The pressure cooker is a great idea! I finished my Myco cleanup efforts a long time ago, but I imagine it'd be perfect for sterilizing driftwood! I'll have to think about picking one up!


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## JusticeBeaver (Oct 28, 2017)

FlyingShawn said:


> Here is the research study your recommendation led me to:
> 
> Do you happen to test for _M. Marinum_ specifically or just _Mycobacteria_ in general? If you happen to find an _M. Marinum_-contaminated culture, is there any chance you'd have the opportunity to culture it and try some of the acetic acid protocols from the mBio article above?
> 
> The pressure cooker is a great idea! I finished my Myco cleanup efforts a long time ago, but I imagine it'd be perfect for sterilizing driftwood! I'll have to think about picking one up!


Marinum would be highly unlikely in a research lab but I think the test is PCR based instead of smear based (I never do it because it's a pain to do). It's usually faster and cheaper to nuke any cell cultures that are going, clean the incubator and move forward from there, rather than treat it. 70% ethanol is effective against most of the species. Marinum is free living though. Most of the mycobacteria live in dirt and the environment so the source could just as easily be from there rather than somewhere else. 

The only issue with the pressure cooker is that they're typically not very large, but you theoretically sterilize rocks in it if you were inclined, but I would never recommend to heat up rocks. If you buy one you'll probably have one of the best kitchen tool you'll ever buy.


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## roadmaster (Nov 5, 2009)

@FlyingShawn
Thank you very much for the link it was very interesting for me.
Is unclear to me how M Marinum diagnosis was arrived at over several mycobacterium strains found to affect fishes/humans .
Many found in natural or processed water's.
(See).. M fortuitum.MFlavescens,M Chelonae,M Gordonae, M Terrae,M trivale,M Diernhoferi,M Celatum,M Kansasii,M Intracellulare,and Marinum .
Nasty stuff to be sure, or to determine definitively.
My own purely unscientific method for treating fishes might be to lace food they like with vinegar,and then freeze small portions assuming (always a bad thing), the fishes would eat it.
May have to disguise the foods a bit with something else they find palatable.
I agree that for possible dip in solution of 6 % to 10 % vinegar against ?? water ,that more info is needed.
Would need to account somehow for residual in water from possible uneaten medicated food, or waste passed by affected fishes.
Possible water change after 12 hours?
Would have no reservations however with cleaning hardscape/ equipment ,with 10 % or 20 % vinegar (amazon).
Would still toss the substrate and begin with newly rinsed substrate.
Still believe vinegar could be of use considering ever increasing resistance by mycobacterium to antibiotics which are more expensive.
My own unscientific research on the matter began with the German blue Ram's I was caring for and what I firmly believe was mycobacterium or Iridovirus and generally poor imported stock.
Thanks again.
Lee.


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## Surf (Jun 13, 2017)

> This is a long scholarly article but it describes how a university zebrafish facility dealt with an M. marinum outbreak. They apparently used hypochlorite solutions along with hot water to sterilize their tanks. Look at page 16 onwards.


Hypochlorite is bleach. Since they didn't want to kill the fish they use 30ppm solution with 10 minute exposure to kill any bacteria on the embryoes. Since you are not trying to keep fish allive you can use a much stronger solution of bleachand more exposure time to kill everything in the tank. It is very effective if the bleach stays in the tank for 6 hours. Then drain and rinse out the tank with a mixture of baking soda and clean water 2 times and then do a final rinsewith just water. 

The baking soda will react with bleach neutralizing the chlorine by converting it to sodium chloride ( table salt). The bleach kills the bacteria by destroying the cell wall. Hikers use bleach, iodine, or chlorine tablet to treat drinking water all the time although it can take 2 hours to kill the some resistant parasites. It also kills algae. Most plastics will handle beach very well and will not be contaminated by it in any way.

Be sure to use bleach without any perfumes or soap in it. These may not be rinsed out as easily as the bleach and baking soda. The university used lab grade bleach to avoid perfumes and soap contamination.

For the substrate and lava rocks put them in the oven on the clean setting (which is over 500F) for several hours. That will covert ant bacteria into dead charcoal. This is 100% effective as long as the entire rock reaches the oven temperature.


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