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Offlinelester
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B+ and colonization
    #7632015 - 11/13/07 06:17 PM (16 years, 2 months ago)

Using the PFTek guide for BRF cakes.

.. got the substrate and injection done on Friday. Today is Tuesday (4 days after), and I can't see much evidence for colonization yet. (unless the whitening look on the BRF is what I'm after .. but it's kind of looked that way since Friday? ..)

Is this normal? That jars are in a dark place at about room temperature. Any need to switch conditions?

TIA.


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OfflineDrewwyann
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Re: B+ and colonization [Re: lester]
    #7632028 - 11/13/07 06:19 PM (16 years, 2 months ago)

completely normal. Some people don't see mycelium growth until a week or sometimes more.

It usually takes 2-5 days to see some growth. So you just have some slow germinating spores is all.

Be patient, you'll see growth soon.


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OfflinePooter
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Re: B+ and colonization [Re: Drewwyann]
    #7632235 - 11/13/07 07:13 PM (16 years, 2 months ago)

BRF is usually pretty slow.


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Offlinelester
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Re: B+ and colonization [Re: Pooter]
    #7632380 - 11/13/07 07:47 PM (16 years, 2 months ago)

Ok, well I wasn't sure if the temp was too high after the jars sterilized (cooled them a bit with a fan).

I'll keep waiting I guess ... thanx.


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OfflineMarlboroMan
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Re: B+ and colonization [Re: lester]
    #7632443 - 11/13/07 08:04 PM (16 years, 2 months ago)

Yeah, once B+ starts showing, you'll be at 100% in no time. My jars with this strain have always colonized really fast. :rockon:


--------------------
I'll make it to the moon if I have to crawl..


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Invisiblethedefone
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Re: B+ and colonization [Re: MarlboroMan]
    #7632462 - 11/13/07 08:10 PM (16 years, 2 months ago)

Quote:

(cooled them a bit with a fan)




They sat around for like 12/24 hours after sterilization, no? If you just cooled them with a fan, and then knocked them up a couple hours later.....

Start Over.


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Offlineblueshroprachaun
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Re: B+ and colonization [Re: thedefone]
    #7632516 - 11/13/07 08:25 PM (16 years, 2 months ago)

Quote:

(cooled them a bit with a fan)
They sat around for like 12/24 hours after sterilization, no?  If you just cooled them with a fan, and then knocked them up a couple hours later..... 
Start Over.



:thumbup:

IME allowing them to sit, foil and lid fully intact, for 36-48 hours is helpful for identifying contams before you waste spores. I would say 24 hours would be the minimum...


--------------------
"All things are poison and nothing is without poison, only the dose permits something not to be poisonous." - Paracelsus,  the father of toxicology


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Invisiblemycocurious
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Re: B+ and colonization [Re: blueshroprachaun]
    #7632603 - 11/13/07 08:48 PM (16 years, 2 months ago)

Quote:

blueshroprachaun said:
:thumbup:

IME allowing them to sit, foil and lid fully intact, for 36-48 hours is helpful for identifying contams before you waste spores. I would say 24 hours would be the minimum...




If the were steam sterilized through a steamer/canner or a pot of boiling water, there may be some merit to waiting that long for some reason.  However, if they were autoclaved or pressure-cooked sterilized, the best thing to do is to turn the power off the unit, leave it where it is, and walk away for 8-12 hours.

It's very important to let the jars cool slowly within the pressurized chamber so that pressures within the chamber (and more importantly, within your jars) equalize slowly and evenly along with the heat generated by the process.  Prematurely depressurizing the unit will cause the vacuum inside to suck contaminated in very rapidly and throughly...all the way into your jars.

Additionally, pressure sterilized or not, the jars will still be terminally hot at the core of the jar well after they feel cool to the touch.  There's a certain practicality to patience...


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.


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Offlineblueshroprachaun
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Re: B+ and colonization [Re: mycocurious]
    #7632672 - 11/13/07 09:04 PM (16 years, 2 months ago)

Quote:

mycocurious said:

It's very important to let the jars cool slowly within the pressurized chamber so that pressures within the chamber (and more importantly, within your jars) equalize slowly and evenly along with the heat generated by the process.  Prematurely depressurizing the unit will cause the vacuum inside to suck contaminated in very rapidly and throughly...all the way into your jars.

Additionally, pressure sterilized or not, the jars will still be terminally hot at the core of the jar well after they feel cool to the touch.  There's a certain practicality to patience...




Very Very Good Point - The only time I ran into contams after PCing them is when I got impatient because I had more jars to do... :mushroom2: :mushroom2:


--------------------
"All things are poison and nothing is without poison, only the dose permits something not to be poisonous." - Paracelsus,  the father of toxicology


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Offlinegreystealth
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Re: B+ and colonization [Re: Drewwyann]
    #7632912 - 11/13/07 09:55 PM (16 years, 2 months ago)

cultivating will probably be difficult for me then, being as impatient as I am. Heh.


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OfflineBlargIAmDead
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Re: B+ and colonization [Re: blueshroprachaun]
    #7632922 - 11/13/07 09:57 PM (16 years, 2 months ago)

Hmmm I've been steam sterilizing mine and I wait only 7-9 hours before I inoculate...could be the reason nothing has grown on my first four B+ jars. Will let sit longer. Thanks for the unintentional advice guys :smile:.


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OfflineMarlboroMan
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Re: B+ and colonization [Re: blueshroprachaun]
    #7632937 - 11/13/07 09:59 PM (16 years, 2 months ago)

Quote:

blueshroprachaun said:
IME allowing them to sit, foil and lid fully intact, for 36-48 hours is helpful for identifying contams before you waste spores. I would say 24 hours would be the minimum...



I do the same thing, hasn't failed me yet! But then again, I steam sterilize. :rockon:


--------------------
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OfflineThe shroomy 1
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Re: B+ and colonization [Re: mycocurious]
    #7633504 - 11/14/07 12:33 AM (16 years, 2 months ago)

Quote:

mycocurious said:
Quote:

blueshroprachaun said:
:thumbup:

IME allowing them to sit, foil and lid fully intact, for 36-48 hours is helpful for identifying contams before you waste spores. I would say 24 hours would be the minimum...






Additionally, pressure sterilized or not, the jars will still be terminally hot at the core of the jar well after they feel cool to the touch.  There's a certain practicality to patience...




I'm kind of confused at this point Myco....  In my studies, I was under the impression that the type of heat transfer we're dealing with is related to conduction.  If that is the case, Heat would spread from the core outward towards the glass jar.  Remembering that things in life don't get "cold", they get "less warm"  The outer glass of the jar should in fact reflect the core temperature until the core and the outer glass equalized to room temperature.  I could be wrong. Just my  .02


--------------------


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Invisiblemycocurious
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Re: B+ and colonization [Re: The shroomy 1]
    #7633910 - 11/14/07 06:32 AM (16 years, 2 months ago)

I'm not going to get backed into a thermodynamics debate but don't forget to consider...

1. glass is an insulator
2. the energy will be lost from the path of least resistance, which is typically through the top/lid as it's not as good an insulator as glass.

and to put a finer point on it, the energy loss is happening through radiation of the heat, not conduction...which in turn causes the outer edges of the jar to pull heat - at a much slower rate - via conduction from the core.

beyond that you got me, I haven't had a physics class in over a decade, I'm just defining observed phenomenon, you can prove it out however you want but I think ultimately it'll come back to the fact that glass is a great insulator of heat. :shrug:


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.


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Offlinelester
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Re: B+ and colonization [Re: lester]
    #7634010 - 11/14/07 07:52 AM (16 years, 2 months ago)

Kind of frustrating then, since the tek I followed had no idication fo time to wait after sterilization.

Guess I'll just wait and cross fingers ..


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Invisiblemycocurious
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Re: B+ and colonization [Re: lester]
    #7634064 - 11/14/07 08:25 AM (16 years, 2 months ago)

:yesnod: 

(this advice is universal in all research you every do...)
Consider every written article to be suspect.  Because it is written does not make it true.  Consider the source (author) and evaluate his attention and how well he communicates his ideas to you as an indication of the validity of his statements.  Understand the meaning of the word "research".  Research is not the process of looking up the answer to a question.  That's searching.  Researching is the process of validating the evidence and information you've discovered with your peers/colleagues and supplementing it with additional data.

I had an english teacher tell me that, god, a couple decades ago now and it really stuck.  This is especially true in the internet age when we have online encyclopedias that are free edited by anyone who decided they're a credible source...
-----------

Look at this situation as a very valuable learning opportunity.  Let the jars sit at room temperature for the next month.  If some spores survived you'll be able to watch them struggle to colonize the entire substrate and if you get any contaminates you'll be able to watch them grow and process full cycle before pitching them.  I think everyone should purposely contaminate a jar with trichoderma, cobweb and a pin mold just to see how they colonize and what they're life-cycles are like.


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.


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Invisibleblackfir
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Re: B+ and colonization [Re: mycocurious]
    #7634092 - 11/14/07 08:37 AM (16 years, 2 months ago)

:werd:


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Tell me, I pray thee, where the seer's house is.


Edited by blackfir (11/14/07 09:34 AM)


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Offlinelester
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Re: B+ and colonization [Re: blackfir]
    #7648963 - 11/17/07 08:40 PM (16 years, 2 months ago)

It's now an even 7 days and nothing .. I'm thinking spores were lost after innoc. ..

o well.


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OfflineAzurescendence
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Re: B+ and colonization [Re: lester]
    #7649381 - 11/17/07 10:36 PM (16 years, 2 months ago)

Just to be safe it's always best (and this is covered in some teks, but not all) to wait overnight (if possible... maybe even PC/boil/whatever them in the evening so you can let them cool while you're sleeping) to be SURE that the residual heat inside the jars is not a potential problem... if you inoculate shortly after a good sterilization, you will, unfortunately, kill off the spores.  Wait until after the jars are cool to the touch even, as I said, many people have said 8-12 hours or overnight just to be safe. Best of luck! :mushroom2:

:peace:


--------------------
~ Peace, love, happiness, and good health to all. ~


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Invisiblethatamatt
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Re: B+ and colonization [Re: lester]
    #7649639 - 11/18/07 12:09 AM (16 years, 2 months ago)

I had the same problem with B+. It took literally 13 days to see some mycelium. I felt like I was watching paint dry. Incubation temp was 72F. BRF, SF-TEK. Just wait a Little longer. If nothing by 14 days there probably dead.


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Offlineveda_sticks
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Re: B+ and colonization [Re: thatamatt]
    #7650048 - 11/18/07 04:58 AM (16 years, 2 months ago)

Some people have not seen germination until as late as 2 weeks, so u will just have to wait, 4 days is quite early, 3 days is the minimum time that germination will happen.

BRF jars are not slow at germination, i have seen germination in my jars in as little as 3 days but generaly is day 6 or 7 before i see anything.

I have inoced 4-5 hours after sterilising and they have worked.

Its best to let your pressure cooker cool for 12 hours just to be safe.

Also remember that if the moisture content is off this can have an effect.


--------------------

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Offlinelester
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Re: B+ and colonization [Re: thatamatt]
    #7652757 - 11/18/07 09:29 PM (16 years, 2 months ago)

Ok - another thing: I re-covered the innoc. holes with tape before putting them away .. does this slow colonization? Do they need some air now?


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Invisiblethatamatt
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Re: B+ and colonization [Re: lester]
    #7653198 - 11/19/07 12:20 AM (16 years, 2 months ago)

There's enough air for now. Leave the tape on until you see the mycelium and then remove. The first time I did not use tape on the holes. guess what happened?  10 out of 12 were infected with greenish mold about the same time the mycelium appeared. It least a got 2 cakes flushing, as I type this.
Good Luck:smirk:


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Offlinelester
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Re: B+ and colonization [Re: thatamatt]
    #7654556 - 11/19/07 12:29 PM (16 years, 2 months ago)

Thanks m8 - I'll keep you posted ..


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Offlinelester
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Re: B+ and colonization [Re: veda_sticks]
    #7658008 - 11/20/07 07:20 AM (16 years, 2 months ago)

Quote:

veda_sticks said:
Also remember that if the moisture content is off this can have an effect.




I can see moisture at the top of the jar as well as notices the whitening in the BRF where I inoculated, so I think it still has a chance ..


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Offlinelester
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Re: B+ and colonization [Re: lester]
    #7667758 - 11/22/07 06:25 PM (16 years, 2 months ago)

Now I see some pinking .. but really sparse .. 13 days now.

Put the jars on front of a heater to warm a bit .. still around 70 deg.

Is there a shelf life for syringes, as these weren't used for 2 months after delivery? ..


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Invisibleblackfir
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Re: B+ and colonization [Re: lester]
    #7668052 - 11/22/07 07:53 PM (16 years, 2 months ago)

\I would say after a few months you should be okay.I've always read to keep them in the fridge until use. I would try to incubate at 78-80 degrees. I think 70's a tad too cold. Pink might not be so good.


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Offlinelester
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Re: B+ and colonization [Re: blackfir]
    #7670322 - 11/23/07 12:33 PM (16 years, 2 months ago)

Well maybe since I'm such a n00b, does the gradual whitening indicate colonization, or is it just moist BRF? I've seen 100% colonization on pics, but it looks more "spongy" than my jars ..


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Invisibleblackfir
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Re: B+ and colonization [Re: lester]
    #7670457 - 11/23/07 12:59 PM (16 years, 2 months ago)

Yes, as long as coming out of the points at which you inoculated. like this here
not my pic BTW. My friends didn't look so white and fluffy either. I think alot of that has to do with the flash. Once out of the jars they look much better. IMO anyway.


--------------------
Tell me, I pray thee, where the seer's house is.


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Offlinelester
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Re: B+ and colonization [Re: blackfir]
    #7674349 - 11/24/07 12:15 PM (16 years, 2 months ago)

Here they are after exactly 2 weeks:




dead?


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Offlinepazzy
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Re: B+ and colonization [Re: lester]
    #7674376 - 11/24/07 12:23 PM (16 years, 2 months ago)

Why did you only fill the jars half full?

You should have filled them to about a quarter inch from the top with pf mix then used dry vermiculite to fill them to the top.

They don't look dead though they are just colonizing really slow.


--------------------
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Offlinelester
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Re: B+ and colonization [Re: pazzy]
    #7674399 - 11/24/07 12:29 PM (16 years, 2 months ago)

Ya, the tek I was following had the mixtures for half pint jars, and these are bigger (had to borrow them). I looked around and it appears using bigger jars is ok.

So, maybe the colonization is slower because of the jar size?


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OfflineMarlboroMan
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Re: B+ and colonization [Re: pazzy]
    #7674401 - 11/24/07 12:30 PM (16 years, 2 months ago)

Quote:

pazzy said:
Why did you only fill the jars half full?

You should have filled them to about a quarter inch from the top with pf mix then used dry vermiculite to fill them to the top.



:thumbup:

What temperature do you have them at? :rockon:


--------------------
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Offlinelester
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Re: B+ and colonization [Re: MarlboroMan]
    #7674412 - 11/24/07 12:32 PM (16 years, 2 months ago)

Had them in front of a heater set at 64 .. now moved it up to 70


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Offlinelester
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Re: B+ and colonization [Re: MarlboroMan]
    #7684870 - 11/27/07 07:54 AM (16 years, 2 months ago)

Would the jars half full stop colonization - of just slow it down?

Should I crack them and add more dry verm on the top?


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Offlinemad genius
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Re: B+ and colonization [Re: lester]
    #7684979 - 11/27/07 08:41 AM (16 years, 2 months ago)

No do not open the jars, from what ive been told its a high risk for contams. The temperature is probably why your mycelium is movin so slow. Ideal incubation temp. is about 80F.


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OfflineBlargIAmDead
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Re: B+ and colonization [Re: mad genius]
    #7685546 - 11/27/07 11:20 AM (16 years, 2 months ago)

Room temp is fine. 70-75 is good. Yes it goes slower than higher temps but bacteria also like higher temps.


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Offlinelester
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Re: B+ and colonization [Re: BlargIAmDead]
    #7694684 - 11/29/07 12:43 PM (16 years, 2 months ago)

Still nothing .. tomorrow is 3 weeks. They've stayed about the same as above 2 pics of jars ..

Damn, do I need a contained chamber at the right temp?


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OfflineKCizzle
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Re: B+ and colonization [Re: lester]
    #7694851 - 11/29/07 01:27 PM (16 years, 2 months ago)

I would def keep them at 82-86. Bacteria likes it but just keep your area nice and clean. 60-70 was most definitely the reason for slow colonization.


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Offlinelester
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Re: B+ and colonization [Re: KCizzle]
    #7695391 - 11/29/07 03:49 PM (16 years, 2 months ago)

The advice is really helpful - thanks.

I actually had them in a thermos container (for picnics), angled sideways towards a heater which blew warm air into the container. At a couple of points, I felt the jars and they were bordering on HOT, so I pulled them out for a while. My room temp fluctuates between 68-70 depending on the time of the day.

So - should I keep them out of the container at 68-70, or in it? (putting them at less peril by keeping them away from the direct blast of the heater fan ..)

Thanks!


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Offlinemad genius
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Re: B+ and colonization [Re: lester]
    #7696167 - 11/29/07 06:22 PM (16 years, 2 months ago)



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InvisibleOptx
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Re: B+ and colonization [Re: KCizzle]
    #7697020 - 11/29/07 10:10 PM (16 years, 2 months ago)

Quote:

KCizzle said:
I would def keep them at 82-86. Bacteria likes it but just keep your area nice and clean. 60-70 was most definitely the reason for slow colonization.




bad. don't incubate over 78. the jars will be just fine


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please do not take everything i say here so personally. welcome to teh internets!


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Offlinemad genius
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Re: B+ and colonization [Re: Optx]
    #7702516 - 12/01/07 11:42 AM (16 years, 2 months ago)

why not over 79?


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OfflineBlargIAmDead
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Re: B+ and colonization [Re: mad genius]
    #7702627 - 12/01/07 12:23 PM (16 years, 2 months ago)

That's the temperature that mushrooms explode at. That and bacteria enjoy higher heat. This way the mushrooms can still grow but we try to inhibit the growth of bacteria.


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Offlinemad genius
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Re: B+ and colonization [Re: BlargIAmDead]
    #7705683 - 12/02/07 07:17 AM (16 years, 1 month ago)

I've heard from about twenty ppl on here that 80F is ideal.


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OfflineBlargIAmDead
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Re: B+ and colonization [Re: mad genius]
    #7708542 - 12/02/07 09:24 PM (16 years, 1 month ago)

So go with what everyone says. The key to success in this hobby is find what works for YOU. Not what works for SWIM, or your cat, or a FOAF, or the President....Doesn't mean anything if it doesn't work under your conditions. It's when you start getting at really high temps like the 100's you have to worry about killing the mycelium and drying the substrate. So rock the casba and do whatever you want. :smile:


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Offlinemad genius
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Re: B+ and colonization [Re: BlargIAmDead]
    #7710193 - 12/03/07 10:26 AM (16 years, 1 month ago)

Well as a beginner, im doing what is suggested by the majority of successful growers. It just sounds weird to me when someone says something contrary to what about 20 experienced growers have said. Especially when this is the advice that I am following.


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OfflineBlargIAmDead
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Re: B+ and colonization [Re: mad genius]
    #7712049 - 12/03/07 05:39 PM (16 years, 1 month ago)

Then stick to your guns Mr. Genius and be damned!! </pirate> Seriously man, 80F is fine for mushrooms. Good luck :smile:.


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Offlinemad genius
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Re: B+ and colonization [Re: BlargIAmDead]
    #7712411 - 12/03/07 06:55 PM (16 years, 1 month ago)

Thanx :wink:


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Offlinelester
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Registered: 05/18/07
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Re: B+ and colonization [Re: mad genius]
    #7719909 - 12/05/07 02:29 PM (16 years, 1 month ago)

Project is scrapped - buried them outside. No change after 4 weeks.


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