Microbe Heat Resistance and Autoclave Testing

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    Heat Resistance
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Discussion Overview

The discussion centers around the heat resistance of microbes, particularly in relation to autoclave testing and validation. Participants explore the effectiveness of autoclaving in killing various microbial species, including those with extreme heat tolerance, and share references and practical advice for ensuring autoclave efficacy.

Discussion Character

  • Exploratory
  • Technical explanation
  • Debate/contested
  • Experimental/applied

Main Points Raised

  • One participant seeks references for heat resistance of microbes, specifically asking for D-values and z values related to autoclave effectiveness.
  • Another participant mentions that certain archaebacteria, like Pyrolobus fumarii, are not killed by standard autoclave conditions, highlighting their extreme heat tolerance.
  • It is suggested that standard autoclaving protocols likely eliminate over 99% of typical contaminating bacteria, but extreme-environment tolerant species may not be addressed.
  • A reference to Fedegari's write-up on F0 calculation is provided as a resource for understanding sterilization processes.
  • One participant shares their experience with spore strips for validation, noting that they found sufficient references to confirm that the microbes on their list are easier to kill than B. stearothermophilus.
  • Another participant points out that testing kits for autoclaves are primarily for operational verification rather than specific bacterial survival assessment.
  • There is mention of tardigrades being able to survive extreme temperatures, which raises questions about the limits of microbial resistance.

Areas of Agreement / Disagreement

Participants express a range of views on the effectiveness of autoclaving against various microbes, with some highlighting specific species that may survive autoclaving while others suggest that typical protocols are sufficient for most applications. The discussion does not reach a consensus on the effectiveness of autoclaving against all potential microbial threats.

Contextual Notes

There are limitations regarding the assumptions made about the effectiveness of autoclaving for all microbial species, as well as the specific conditions under which certain microbes may survive. The discussion also reflects varying levels of expertise among participants regarding microbial heat resistance.

jamesrc
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Hi,
Does anyone know of a good reference for heat resistance of microbes? What would really help would be any listing of decimal reduction times (D-values) at a given process temperature and z values. I'm just trying to make sure our autoclave kills everything it's supposed to for someone, so I've got a list of microbes with initial # of colony forming units from some bioburden studies. If anyone can point me towards a good reference, that would be a big help. Thanks.
 
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Autoclave doesn't kill certain archaebacteria

Edit: Ok I dug up some info for you:

Pyrolobus fumarii holds the current record for heat stability, with a 106 deg C temperature optimum and 113 deg C maximum - it is so heat tolerant that it is not killed by one-hour treatment in an autoclave (121 deg C)!

Also there are pH-tolerant archaebacteria that grow in highly acidic (pH=0.7) and highly basic (ph=11) environments.

And then there are pressure-tolerant archaebacteria that have been isolated from ocean depths and requiring pressures of 300 atm just to survive, and tolerating up to 800 atm.
 
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In terms of practical use of autoclaving for the destruction of your typical contaminating species I would guess that the standard protocols remove 99%+ of the bacteria on your surfaces. Unless you, or your colleagues, are directly working with them, the chances of you running into any of the extreme-environment tolerant species is minimal.
If you are looking for some sort of laboratory-based testing or verification to reach some clinical threshold you might try a company like http://www.namsa.com/index.asp" for you to test the efficacy of your autoclave. You can either try some of their products or maybe call their tech support for some free advice.
 
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Fedegari has an excellent write up on F0 calculation. You can access the document at http://www.fedegari.com/techno.asp#fo

I read somewhere that Stain121, which generally stays in the ocean depths, can resist the moist sterilization temperatures of 121 deg.C
 
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Thanks for all of the help everyone. We had tried using spore strips for the validation, but the FDA wasn't satisfied. I found a few books that were helpful, especially Thermobacteriology in Food Processing by Stumbo. I think I've managed to find enough references to show that all of the microbes on my list are easier to kill than B. stearothermophilus, so I'm OK. Then again, I don't know jack squat about microbes and had never even heard of an autoclave a month ago. Thanks again.
 
quark said:
Fedegari has an excellent write up on F0 calculation. You can access the document at http://www.fedegari.com/techno.asp#fo
I read somewhere that Stain121, which generally stays in the ocean depths, can resist the moist sterilization temperatures of 121 deg.C

And tardigrades can survive +150 degrees without water,they are pretty much indestructible as bacterias.
 
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Usually the testing kits for autoclaves are intended to detemine if your autoclave is actually working properly, not whether it is supposed to kill certain bacteria at a particular setting.

If you're working with a specific bacteria that you're concerned about surviving extreme environments, I would think the safest approach would be to autoclave media known to contain the bacteria, and then culture it after autoclaving to see if they survived.
 

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