Spore cleaning without agar or centrifuge (pasteurized spores)

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Stipe-n Cap
Full retard

Registered: 08/04/12
Posts: 9,113
Loc: Canada

Spore cleaning without agar or centrifuge (pasteurized spores)

23
#29130723 - 02/18/25 08:05 PM (1 year, 7 months ago)

Spores can survive 160°F for 36 hours: Spore revival from desiccated fruits , which means spores can survive pasteurization temperature.

If spores survive pasteurization temps, that means dirty syringes can be submerged in a pasteurization bath or run through a dehydrator to neutralize most pesky contaminants, thereby making them more reliable as inoculum. Infected prints can be loaded into syringes, vials, or jars, to be pasteurized in the same fashion. This is a great opportunity to play with those known contaminated syringes or prints that've been giving you a headache, and to break out your sous vide. :lol:

Depending on how effective this process is, we may be able to eliminate the necessity for agar when directing new growers how best to get a foot in the door. This will obviously not replace culture work, for obvious reasons, no matter the level of effectiveness.

I've proven the concept for cube spore survival at 160°F at least, hyperlinked above. We know vegetative bacteria, yeasts, and molds die at pasteurization temperatures, obviously, otherwise pasteurization wouldn't work; so, with these basic facts combined it seems reasonable we could get away with this. Dropping the temp down from 160°F to 145-150°F should provide a greater likelihood of active spore survival.

145°F for 60 mins will be my recommended minimum basiline, with adjustments made if necessary. The sous vide/dehydrator max temp should be between ~ 185-195°F (85-90°C), lot's of room for adjustment. We're not looking for sterility, just a cleaner more reliable starting material. I'm not sure what the upper thermal limit is for active species spore survival but, we can get it figured out. Should you not own a sous vide or dehydrator, regular stovetopor hot plate pasteurization with a thermometer will be the next best option.

Keep in mind we don't know how many of our target species spores die off when exposed to these temperatures, this is why I'm suggesting an initially short exposure, and low temperature. This will definitely be a consideration when attempting to preserve heterozygosity with wild spores, So, what we should be focusing on is who can get the best results at the lowest temp and exposure time. I wouldn't go lower than 30 mins, but 1 hour seems reasonable.

Remember we're attempting to pasteurize, not sterilize. Higher temperatures means higher likelihood of destroying the very spores we're trying to salvage. Our aim is to kill vegetative bacteria, mold, and yeast. The likelihood we'll incubate any endospores is unlikely, as they shouldn't be present, but even if they were, they're not what we're concerned about here.

The objective is to destroy heat sensitive vegetative competitor organisms while inflicting minimum damage on our active spores. Having said all of that, there will be absolutely zero difference between 145°F and 160°F, or even 190°F where endospores are concerned, they might as well be the same temperature at these exposure times. Endospores will not reactivate without a growth medium even at preferred incubation temperatures.However, all data points are valuable so even if someone chooses longer exposure at higher temps, the results will help plot and dial our preferred outcome.

Granted, the proof of concept was done with dry heat but I doubt the difference between wet and dry heat will matter much at these temps; all we need noe is for enough people to test the hypothesis.
My cult equipment is in storage and I don't currently have any spore stock. I'm posting this for the community to play with.

Post any results here.

Extras:

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