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Offlinejason9086
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Contamination on rye berries *DELETED*
    #25195912 - 05/10/18 03:51 PM (6 years, 8 months ago)

Post deleted by jason9086

Reason for deletion: Shared too much personal information

Edited by jason9086 (05/10/18 03:52 PM)

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InvisiblebodhisattaMDiscordReddit
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Re: Contamination on rye berries [Re: jason9086]
    #25196025 - 05/10/18 04:38 PM (6 years, 8 months ago)

rye takes at least 90m at 15psi(121C) some people like to do 2 hours.
if you soaked it 3 days first you loaded it up with bacteria(may not smell or appear like it) which makes it harder to sterilize.

what went wrong? basically everything unfortunately

autoclave tape is useless. all it does is change color when exposed to 121C steam. this doesn't mean anything is sterilize it only means something has been exposed to the inside of a sterilizer. they use autoclave tape in industry and medical so that things don't get autoclaved twice. not to verify if the sterilization was successful. To validate sterilization you need to use a biological indicator.

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Offlinejason9086
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Re: Contamination on rye berries [Re: bodhisatta]
    #25196043 - 05/10/18 04:51 PM (6 years, 8 months ago)

Thanks,

The autoclave tape I mentioned was not to imply that the sterilization was successful. Obviously it wasn't. I am new to cultivating mushrooms and sterilizing non-liquid materials, but I have worked with conidia-forming ascomycetes and preparing media for culturing them. The point of mentioning this was simply to say that I didn't simply leave them open in between the autoclave and the laminar hood.

It sounds like my sterilization time was too low based on your information. I am a little surprised that people would do up to two hours. Disposing of even regulated organisms, it is only necessary to autoclave for 60 mins at these temps.

Thanks for the input

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Offlinejason9086
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Re: Contamination on rye berries [Re: bodhisatta]
    #25196050 - 05/10/18 04:55 PM (6 years, 8 months ago)

I just checked the autoclave settings. It was actually at 18psi, not 15. The temp was set to 121c. I think I should have done longer than 30 mins for sure, though

Pretty sure its an Aspergillus species

Edited by jason9086 (05/10/18 05:01 PM)

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Re: Contamination on rye berries [Re: jason9086]
    #25196147 - 05/10/18 05:41 PM (6 years, 8 months ago)

You need more time for the temperature to heat through. I doubt your autoclave is designed for sterilizing 5 lb bags of rye.

I always sterilize at 15 psi for 2 hours minimum.

Definitely don't go below 90 minutes.

You might get away with 30 if you were just sterilizing agar or maybe a small flask of grains. In this hobby 90 minutes to 2 hours at 15 psi is standard or 8 hours at 100C.


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Edited by elasticaltiger (05/10/18 05:42 PM)

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Re: Contamination on rye berries [Re: elasticaltiger]
    #25196159 - 05/10/18 05:45 PM (6 years, 8 months ago)

grain is far harder to sterilize than tools and liquids. grains trap air and air is hard to remove and air is the no1 reason for failed sterilization. let alone the fact that they're just plain hard to sterilize being organic nook and cranny objects. vs say a scalpel or some used scrubs or agar.

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Offlinejason9086
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Re: Contamination on rye berries [Re: bodhisatta]
    #25196294 - 05/10/18 06:49 PM (6 years, 8 months ago)

Dumping all of em and retrying.

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