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Invisiblepsyphon
mneumatic device

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Registered: 11/27/01
Posts: 565
Help with stalled cake colonization
    #479933 - 12/04/01 07:50 PM (22 years, 3 months ago)

My pet Ling Chih wanted some cubie friends so he tried the PF and dunk teks and has had significant success but has recently run into some problems. Here is the current situation: 4 out of 6 jars are surviving in an incubation chamber constructed out of a sterilite container, water, hardware cloth to hold the jars above the water level and a heating pad. The temperature is 80-87 whenever the jars are checked. H2O2 and ethyl rubbing alcohol are occationally added to the water to ward off contams. It has been six weeks since inoculation and the jars seem to have halted growth at about 75% since the third week. At 50% the jars were turned upside down and the tape removed from the holes. Also in some of the jars I have noticed something strange...the myceilium has turned an off-white color and then bright white patches have grown over the darker ones but have not grown further into the uncolonized substrate. WTF?

What should my Ling Chih friend do? It seems to me he has three choices: 1. Using sterile procedure (oven tek and common sense) remove the substrate and case only the colonized portion of the cakes and spraying with water/H2O2 solution.

2. Using sterile procedure remove and case the entire cakes, mixing the colonized with the uncolonized and spraying with water/H2O2 solution.
Or
3. Sterilizing a syringe filled with honey-water and injecting it into the jars in the hopes that this will encourage complete colonization.

Thank you for reading this long post, any help is appreciated.

P.S. What services/programs do you all use to post anonimously? Are any free? Thanx for any info.


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"The real voyage of discovery consists not in seeking new landscapes but in having new eyes."
- Marcel Proust

I wish you all ceaselessly flowing moments of happiness.

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OfflineFreakShow
3n+hu$|@$+
Registered: 11/14/01
Posts: 439
Last seen: 21 years, 3 months
Re: Help with stalled cake colonization [Re: psyphon]
    #479958 - 12/04/01 08:02 PM (22 years, 3 months ago)

what did you use for your substrate mixture? Sounds like you may not have put in enough water.

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Invisiblepsyphon
mneumatic device

Folding@home Statistics
Registered: 11/27/01
Posts: 565
Re: Help with stalled cake colonization [Re: FreakShow]
    #479992 - 12/04/01 08:19 PM (22 years, 3 months ago)

1/4 cup brf
1/2 cup verm
~40cc water
per each 1/2 pint jar.

Problems occured in the past with colonization being slow from too much water when 55cc was used so it was reduced to ~40. Also being in a very humid environment (the incubation chamber) I figured they'd lose little moisture.


--------------------
"The real voyage of discovery consists not in seeking new landscapes but in having new eyes."
- Marcel Proust

I wish you all ceaselessly flowing moments of happiness.

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OfflineFreakShow
3n+hu$|@$+
Registered: 11/14/01
Posts: 439
Last seen: 21 years, 3 months
Re: Help with stalled cake colonization [Re: psyphon]
    #480002 - 12/04/01 08:28 PM (22 years, 3 months ago)

well two things would be most likely slowing colonization of the jar, 1st would be not enough water in the substrate mixture, and 2nd would be lack of air in the jar. I'd up the water to 45-50 cc and you can also try turning the jars upside down for a day. (tap them to make the substrate fall down to the lid.)

actually, i am in the middle (still boiling) of making 1/2 ping wm jars. I'm experimenting with a couple sets of 5 jars w/ various mixtures (mostly water amounts). I logged everything and i'll let ya know how they turn out.hopeuflly i'll be getting the spores from sporeworks by friday.

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Invisiblepuscle
genius of love
Registered: 01/06/01
Posts: 4,539
Loc: NY
Re: Help with stalled cake colonization [Re: psyphon]
    #480687 - 12/05/01 12:50 PM (22 years, 3 months ago)

They're most likely too dry. I would cut away the uncolonized substrate and case. Don't inject honey water. If you want to experiment, inject sterile water.

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Offlinejonnyshaggs420
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Registered: 08/08/00
Posts: 1,965
Loc: Mid-West
Last seen: 18 years, 5 months
Re: Help with stalled cake colonization [Re: psyphon]
    #480706 - 12/05/01 01:01 PM (22 years, 3 months ago)

Could be lack of water. 45-50 cc's is a good mix for the pf tech, but it all depends on the substrate.

Could also be the addition of alcohol to the water. The alcohol may have evaporated and upon entering into the jars killed the mycelium (this is just a theory)

You could also have a bacterial contamination, which is sometimes a problem with humid incubators.


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Invisiblepsyphon
mneumatic device

Folding@home Statistics
Registered: 11/27/01
Posts: 565
Re: Help with stalled cake colonization [Re: jonnyshaggs420]
    #480904 - 12/05/01 03:50 PM (22 years, 3 months ago)

Thank you everyone for your replies. Since the incubation chamber seems like it might be the problem. (Success was had when the weather was warm enough to not need one.) What kind of chamber can beI used that will keep the jars around 80deg and be sterile enough to provide safe gas exchange without the cakes getting dried out? Or is there something that can be done to be sure the water stays sterile enough? I was thinking of adding large amounts of salt or something similar that should prevent microbial life from growing but will not evaporate into the chamber.

Thanks again for your help.


--------------------
"The real voyage of discovery consists not in seeking new landscapes but in having new eyes."
- Marcel Proust

I wish you all ceaselessly flowing moments of happiness.

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