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 Teknion
Interloper
Registered: 02/28/03
Posts: 110
Last seen: 20 years, 2 months
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 BRF Colonization Problem.... 
#2144095 - 11/29/03 10:14 AM (20 years, 4 months ago) |
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So, AFOAF gave me this scenario to post, and wants to know what might be going on....
1) On two separate sessions, he made some BRF jars. He labeled the jars from A-G on the first batch, then H-N on the second. Both batches were PC'ed for approx 80-90 mins.
2) He cleaned his kitchen, lysoled and cleaned everything with alcohol wipes, etc etc.
3) Using 2 new syringes, with the needles still sealed, he innoculated the jars, with the top off. There was a quarter inch of verm to protect the cake itself.
4) The first syringe did 90% of the jars, and then he used the second syringe to finish off the rest. Once done, he added 1/4 of a CC to the middle of each cake to finish the syringe off... This part was done with the tops on since his jars had a hole in the middle.
5) He placed all the jars in an incubator, using water, a tank heater and a bubbler to make the surrounding bin to 84 degrees (bins are not see through). And he let that sit for 14 days.....
6) Since the 6th day, jars F and G have been showing really nice mycelium growth. But all other jars have done nothing in 14 days.
He was just wondering how long he should wait before calling the other jars a write off?
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"A drug person can deal with things like seeing they're dead grandmother crawling up they're leg with a knife in her teeth but no one should be asked to handle this trip"
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 soochi
Chef
Registered: 08/13/02
Posts: 2,420
Loc: The Richest County
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 Re: BRF Colonization Problem.... [Re: Teknion]
#2144147 - 11/29/03 10:45 AM (20 years, 4 months ago) |
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First, you could have added too much water (spore solution), second, when you said you opened the jars, you could have introduced contams (smell the jars that don't show coloniztion) Did you drill holes in the lids? Did you flame sterilize your needle and wipe it down with alcohol after each injection? did you stick the needle into the rice cake? I usually drill four holes around the circumference of the lid (making sure not to drill through the rubber seal) and when injecting, staying as close to the sides of the jars as possible.
Give the jars a couple more days, if they start to smell then chuck em.
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Wee, sleeket, cowran, tim'rous beastie,
O, what panic's in thy breastie!
Thou need na start awa sae hasty,
Wi' bickering brattle!
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Teknion
Interloper
Registered: 02/28/03
Posts: 110
Last seen: 20 years, 2 months
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Re: BRF Colonization Problem.... [Re: soochi]
#2144187 - 11/29/03 11:06 AM (20 years, 4 months ago) |
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I flame sterilized and wiped at every few jars....
When I innoculated the center, yes I did stick the needle into the cake.... For the other innoculation I stayed as close to the glass as possible.
I'm using brittle plastic tops, so I couldn't drill on the sides unfortunately. But the kitchen was uber-clean.
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"A drug person can deal with things like seeing they're dead grandmother crawling up they're leg with a knife in her teeth but no one should be asked to handle this trip"
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GregHimself
Just a guyobssesed withshroomies.
Registered: 07/18/03
Posts: 936
Loc: British Columbia, Canada
Last seen: 6 years, 3 months
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Re: BRF Colonization Problem.... [Re: Teknion]
#2144194 - 11/29/03 11:11 AM (20 years, 4 months ago) |
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How long did you allow the jars to cool before inocculation?
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"The key to successful cultivation is to match the skills of the cultivator with the right strain on the proper substrate under ideal environmental conditions."
Paul Stamets
Growing Gourmet and Medicinal Mushrooms(Third Edition)
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Teknion
Interloper
Registered: 02/28/03
Posts: 110
Last seen: 20 years, 2 months
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Re: BRF Colonization Problem.... [Re: GregHimself]
#2144207 - 11/29/03 11:22 AM (20 years, 4 months ago) |
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The batch from A-G was cooled for several days....
H-N was cooled for 10-11 hours.....
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"A drug person can deal with things like seeing they're dead grandmother crawling up they're leg with a knife in her teeth but no one should be asked to handle this trip"
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Teknion
Interloper
Registered: 02/28/03
Posts: 110
Last seen: 20 years, 2 months
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Re: BRF Colonization Problem.... [Re: Teknion]
#2144213 - 11/29/03 11:27 AM (20 years, 4 months ago) |
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I'm wondering if it could be this.....
He had covered the middle holes with masking tape. After he was done PC'ing.... And started to innoculate, he noticed the middle of the verm layer was slightly damp, indictaing some moisture crept in during pc'ing.... But it didn't look too bad....
But now Im thinking he may have been wrong.....
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"A drug person can deal with things like seeing they're dead grandmother crawling up they're leg with a knife in her teeth but no one should be asked to handle this trip"
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turtlelamp
n00b
Registered: 11/27/03
Posts: 93
Loc: The Midwest
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Re: BRF Colonization Problem.... [Re: Teknion]
#2144256 - 11/29/03 12:04 PM (20 years, 4 months ago) |
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I doubt it was too much water. I've had jars that were saturated to the point of being muddy that completely colonized (although I wouldn't recommend it.) However, I have been struggling with a similar problem myself. Do your jars themselves come in contact with the water in the incubation chamber? That's a no-no. The relative humidity around the jars is much too high and this seems to inhibit colonization... I'm not certain why, but I'm trying to prove this right now through a controlled experiment. My jars, however, indicate an unwillingness to germinate by generating a lurid, bacterial stench.
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Fingerprints on file.
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Magash
Da Bud Guru
Registered: 07/25/02
Posts: 5,876
Loc: Near Hilo
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Re: BRF Colonization Problem.... [Re: Teknion]
#2144363 - 11/29/03 12:57 PM (20 years, 4 months ago) |
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Next time try PC ing for way less time.
Grow Log
Tek : PF
Strain : South American [SA]
Spores : Came from homemade prints. 36 total jars to be injected.
Substrate :Brown Rice Flour [BRF] and Vermiculite [Verm]
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10:15 PM 3/4/03 : Project is started. Jar prep is just putting small holes in the lids for the spore syringe and then taping them closed until inoculation. Tape used is the standard black electrical tape. First batch of jars is mixed. The mix used is 2 parts verm, 1 part water, and 1 part brf. [ This mix is the max fruiting formula by PF.] Mix is put into 1/2 pint canning jars lightly, not packed in, and left it airy. Put substrate in until it reaches the threads of the jar, then add dry verm the rest of the way.
Jars are PCd at 10psi or 240f for a period of 45 minutes. Then jars were left to sit overnight to cool.
8:39 PM 3/5/2003 : First batch of jars injected. (24 jars)
10:04 AM 3/6/2003 : Incubator is running to low 77F. Going to turn it up today.
5:45 PM 3/6/2003 : Second batch of jars is in the PC will be injected tomorrow. Turned up incubator runs at 83F. 86F is the best and if the incubator is 83F and myc growing in the jars is suppose to create a little heat I figure this should work out well.
1:11 PM 3/7/2003 : Second batch of jars is done. (12 jars)
12:10 PM 3/9/2003 : Jars are doing good, no signs of contams
2:54 AM 3/19/2003 : Tape removed from the holes today and coffee filters put over the top of the jars for added protection. Holes were opened so the CO2 in the jars could escape. I gave 12 jars to a friend who kept fucking up. 24 jar?s about 75%done.
12:13 PM 3/22/2003 : Well it's Saturday and the jars will probably be done by the 25th. So it was 36 jars. 12 were given to a friend, 24 done by Tuesday.
10:26 PM 3/23/2003 : 15 jars were done enough to put into dunk. I dunk and cold shock before birthing and after every flush. The other 9 jars have a spot the size of a pea that has to be covered so I'll do those on the 25th.
7:17 PM 3/24/2003 : 15 jars put into the pod. Put in and put a layer of moist verm on top. With geolite I just put the cakes on it. With some other chambers I put the cake on a lid filled with verm. This is done to give the shrooms on the bottom and the top of the cake some added moisture.
The other 9 jars just finished so I'm going to give them a couple more days to make sure the middle of the cake is done.
4:57 PM 3/27/2003 : 5 More cakes birthed today, 4 left going to do them tomorrow or 3/29/2003
3:58 PM 3/29/2003 : Well I lost another cake. My wife's little 1 cake pod made from a 2 liter and a small fish pump and a little geolite used up her last cake so I gave her a SA to grow. The last 3 are in the cold shock dunk.
So the total is 36 jars done all colonized with no contams. 12 given to a friend, 1 to the wife, 23 mine
12:08 PM 3/30/2003 : Last of jars put in
12:02 PM 3/31/2003 : The first 15 cakes are pinning nicely and some have shrooms coming from the bottom.
11:01 AM 4/3/2003 : Things look to be going well.
1:02 AM 4/4/2003 : Took the first flush from four of the cakes. More will be done later today.
5/8/2003 : Total 23 cakes 164 grams dried.
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All creatures tremble when faced with violence. All creatures fear death, all love life. If we can only see ourselves in others, then how could we possibly hurt another creature?
 Join us at the Growery!
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GonzoCool
Registered: 09/30/03
Posts: 726
Loc: 
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Re: BRF Colonization Problem.... [Re: Magash]
#2144487 - 11/29/03 01:40 PM (20 years, 4 months ago) |
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Magash, for those, were you just using coffee filters on top? or was that some other material? isnt that to let the breathe after theyre a good bit colonized, without risking contams?
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Magash
Da Bud Guru
Registered: 07/25/02
Posts: 5,876
Loc: Near Hilo
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Re: BRF Colonization Problem.... [Re: GonzoCool]
#2144841 - 11/29/03 04:22 PM (20 years, 4 months ago) |
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Naw, the verm handles that. I just use them for a little added security and dust covers.
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All creatures tremble when faced with violence. All creatures fear death, all love life. If we can only see ourselves in others, then how could we possibly hurt another creature?
Join us at the Growery!
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weedies
Stranger
Registered: 11/15/03
Posts: 19
Last seen: 20 years, 4 months
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Re: BRF Colonization Problem.... [Re: Teknion]
#2145030 - 11/29/03 05:40 PM (20 years, 4 months ago) |
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DUDE, Ive done this before.... you said you PC'ed them for 80-90 minutes? way to long bro. brf only needs 25 minutes at 15 psi. you destroyed the nutrients in the brf.
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weedies
Stranger
Registered: 11/15/03
Posts: 19
Last seen: 20 years, 4 months
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Re: BRF Colonization Problem.... [Re: weedies]
#2145038 - 11/29/03 05:43 PM (20 years, 4 months ago) |
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forgot to say something. When you overcook the brf, some of the jars will colonize. but you wont get any production out of those cakes. I had 10 cakes like that in a Hydropod with perfect conditions, and only produce like 5 mushrooms total because of the lack of fungi food.
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soochi
Chef
Registered: 08/13/02
Posts: 2,420
Loc: The Richest County
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Re: BRF Colonization Problem.... [Re: Teknion]
#2145397 - 11/29/03 08:24 PM (20 years, 4 months ago) |
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Bingo! did you cover the holes after pcing? it should be done before and the lids covered completely with foil to prevent moisture from seeping in. Too much moisture will hinder growth and harbour contams, if not now then definitely later on.
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Wee, sleeket, cowran, tim'rous beastie,
O, what panic's in thy breastie!
Thou need na start awa sae hasty,
Wi' bickering brattle!
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trev
comming out of retirement
Registered: 04/02/03
Posts: 871
Loc: Aussie
Last seen: 12 years, 11 months
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Re: BRF Colonization Problem.... [Re: weedies]
#2147777 - 11/30/03 07:53 PM (20 years, 4 months ago) |
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Is that true? about the over sterilizing coz I use fractional sterilisation 1/2 hr then next day 1/2 then the next 1 hour and all seems well but I have not got a Pc yet.
Oh yah magash my sisters brothers mothers son has spent a bit of cash on the poor mans pod but its not set up yet.
May need some advice some time.
Cheers
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Anonymous #14
[quote]There are billions of people on this planet. The world does not revolve around the united states, moron. I hope terrorists crash their collective cocks into your asshole. [/quote]
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