Which Antibiotics can be autoclaved

[Mega]

Hi everybody, 

for our GFP - plant project at the AEC we will need Kanamycin, Tetracyclin and Streptomycin. 

AFAIK, you can autoclave Kanamycin (but never ever ever Ampicillin! ;) ). What about Tetracyclin and Streptomycin, will they degrade @ 121°C and 2 bars? 

Best, 

Andreas

[Dakota]

I don't know if they will degrade but is there a special reason why they need to go through the autoclave?  Just add them to the media and swirl around before you pour the plates, or whatever it is you are pouring.

[Andreas Sturm]

Ok, thanks. A friend of mine ment he would rather autoclave it. 

Happy new year everybody!

[Brian Degger]

You can filter sterilise using syringe and 0.22 filter.

On 1 Jan 2013 10:25, "Andreas Sturm" <masters...@gmail.com> wrote:

Ok, thanks. A friend of mine ment he would rather autoclave it. 

Happy new year everybody!

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[Andreas Sturm]

Yeah, thanks.

But his intention was to put everything into the erlenmeyer flask and autoclave it all to get it totally aseptical.


Actually, the ethanol will kill most/all contaminants anyway...

[Nathan McCorkle]

A good reason to develop a list of antibiotics that can be autoclaved is dual-use... you also know what antibiotics you can autoclave to DEACTIVATE them (i.e. disposing used media) 

On Mon, Dec 31, 2012 at 10:54 AM, Dakota <dko...@gmail.com> wrote:

I don't know if they will degrade but is there a special reason why they need to go through the autoclave?  Just add them to the media and swirl around before you pour the plates, or whatever it is you are pouring.

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[Dakota]

That's true, and something I hadn't thought of outside of ampicilin.

[Andreas Sturm]

An advantage is easier handling and, once autoclaved, you have a perfectly aseptic, yet unopened bottle of LB-antibiotic.

Perfekt for DIY activities, where you might not have (cheap) acces to filter sterilization device.

[Josiah Zayner]

Usually the point of adding antibiotics is to kill bacteria.
I always add antibiotics after autoclaving and in my almost 10 years and growing thousands of liters of cultures have never had a problem.
In a pinch one can even forgo autoclaving and just add antibiotics(have definitely done this when in a time rush).

[Andreas Sturm]

Never heard you can forgo autoclaving, but it does make sense.

Some hard-core antibiotics kill quite everey bug (be it eucaryotes or prokaryotes). But of course there's the danger of having one contaminant who is naturally resistant.

[Cathal Garvey]

Depending on the Antibiotic, this may be more or less of a good idea. If the antibiotic is bacteriostatic, then it'll only delay the growth of contaminants, so you should autoclave and add filter-sterilised antibiotic; this is the case with Ampicillin.

However, for Kanamycin, you can probably get away with adding directly, as it's bacteriocidal. However, it still makes sense to autoclave the medium prior to adding the antibiotic, or even simply boil it in the microwave briefly, to kill wild resistant bacteria. It's a nice shortcut, but adding directly may still allow wild colonies to grow.

As innocuous as it sounds, that can really mess up an experiment if you take it for a positive result. I've wasted weeks on broths I thought were sterile when pure chance yielded a few colonies on the experimental plates and not the controls.

On 5 January 2013 08:50, Andreas Sturm <masters...@gmail.com> wrote:

Never heard you can forgo autoclaving, but it does make sense.

Some hard-core antibiotics kill quite everey bug (be it eucaryotes or prokaryotes). But of course there's the danger of having one contaminant who is naturally resistant.

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[Josiah Zayner]

Agreed. I wasn't advocating shortcuts just telling my empirical experience. However, I have never worked or seen in an academic lab someone filter sterilize an antibiotic mixture. Considering the price of filters these days that seems like a big waste of money.
Antibiotics are stored at -20 and kill or slow growth of bacteria. Usually many also in 70 % ethanol. I hardly see a need for filter sterilization but each person needs to find their own level of comfort in how strict they need to be in their experimental protocol.

[Cathal Garvey]

For most, probably not an issue, but I have teh rage against Ampicillin for being such an awful antibiotic in general. It's only bacteriostatic for about 48h at 30/37 C and beta-lactamase generates large zones of clearance within ~2 hrs, so contaminating bacteria can easily screw up an experiment.

Worse, in liquid culture, contaminants can actually build up serially from culture to culture, so that the proportion of contaminants to desired cells can grow and grow with each generation. With unstable plasmids, this can be a real killer. I've actually lost plasmid stocks thanks to this; hence my nerd rage.

For Kanamycin, I have suffered less trouble, something I attribute to it killing contaminants at the outset of culture, rather than just delaying their growth.

Honestly, I haven't used other antibiotics in years, so I don't know, but as you say; many are stored in Ethanol, which is pretty effective against vegetative or dormant cells, and may be bacteriocidal to boot. Chloramphenicol comes to mind. Incidentally, Chloramphenicol can be autoclaved, and can't be reliably heat-inactivated.

[Cuong Le]

I know this is late, but I was looking for thermal degradation data myself and after I found it I figured I would post it here for anyone else that stumbles on this thread:

Heat stability of the antimicrobial activity of sixty-two antibacterial agents

J. Antimicrob. Chemother. (1995) 35 (1): 149-154.

doi: 10.1093/jac/35.1.149

Ampicillin and tetracyclin are temperature sensitive; kanamycin is heat stable. They did not include streptomycin; however, US patent 2473339 says: "prior to the present invention it was generally known that it was impossible to autoclave solutions of streptomycin and that such solutions were injured by temperatures materially above 75° Fahrenheit" so yes/no for streptomycin but you might not be able to prepare your solutions in the way that they are making them.

[Andreas Stuermer]

oh man. I miss sci-hub^^

On Fri, Dec 6, 2013 at 1:14 PM, Andreas Stuermer <masters...@gmail.com> wrote:

Thanks, very ineresting!!

Kanamycin heat resistant? Didn't hear of that before. Activity remaining after autoclaving 98% ?

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[Andreas Stuermer]

But thanks, very interesting.

Couldn't remember that Kan was autoclavable, but then looked up my first post :D

[Nathan McCorkle]

On Fri, Dec 6, 2013 at 1:47 AM, Cuong Le <cuong...@gmail.com> wrote:
> I know this is late, but I was looking for thermal degradation data myself
> and after I found it I figured I would post it here for anyone else that
> stumbles on this thread:
>
> Heat stability of the antimicrobial activity of sixty-two antibacterial
> agents
> J. Antimicrob. Chemother. (1995) 35 (1): 149-154.
> doi: 10.1093/jac/35.1.149

Thanks for the great reference!

Here:
http://diyhpl.us/~nmz787/pdf/Journal_of_Antimicrobial_Chemotherapy_1995_Traub_149-154.pdf


So I'd think the one's that are heat-labile should be at the top of
the DIY usage list, so any discarded media wouldn't contain any active
antibiotics to aid selection for potential DNA-induced horizontal
transfer. Maybe labs should have some DNase on hand too, or maybe that
should be engineered in a organelle that will do some clean-up on cell
death (programmed or otherwise).

[Xabier Vázquez Campos]

Kanamycin and chloramphenicol are some of the few heat-resistant antibiotics. Indeed, kanamycin-resistance genes are used for selection of transformed cells in studies with Thermus spp. One of the few antibiotics suitable to work with thermophiles