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Offlineaktar
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Registered: 02/27/11
Posts: 58
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first grow... off to a bad start
    #14038275 - 02/27/11 08:19 PM (12 years, 10 months ago)

First of all, hello — good to join you all here on Shroomery.


Check out my latest grow here.
Keep up to date with my grow log.



I have been familiar with this site for a while and finally decided to sign up because I need some good old advice from the experts. I got my hands on a few syringes; after rounding up materials, I decided to get some wide-mouth jars, load them up with substrate and inoculate them.

the substrate was made with the following (for 9 pint jars):
— 9 c. horticultural vermiculite
— 4.5 c. brown rice flour
— 4.5 c. water
— a couple tsp. bee pollen + nutritional yeast (suggested here)

these were mixed homogeneously and the mixture was loaded into jars. I checked the wetness and it was damp (felt kind of wet when you squeezed it, but no water came out). the jars were topped off with verm and closed up. the lids had 3 inoculation sites drilled into them, which were covered with 3m transpore medical tape (i wasn't sure how well it would substitute for micropore, but thats what had).

the jars were pasteurized in a water bath for 45 minutes. seven of them were inoculated with MS. the syringe was flame sterilized and swabbed with isopropyl with each injection, then was pushed through the tape, as deep into the jar as far as it could go, angled just slightly outward, and 1cc was injected for each site (3cc per pint jar). Was this too much?

the jars were kept wrapped in a blanket with a heating pad, temp between 68-85°, the vast majority of the time it was 76-82°F. If cubies can't tolerate fluctuating temps, then this might have been an issue. I was probably not patient enough (had to check them frequently), and the jars have seen consistent, intermittent light over the past few weeks.

Five days after inoculation I started to see signs of growth. Small, whitish spots began to appear on some of the jars:


Oh, the joy in my heart when I saw the first indication of success! Two weeks after inoculation, most of the jars were coming along at a very slow pace. One however, was very promising:


3 weeks and after inoculation, I had little hope for 5 of the jars. But two were coming along, and particularly the champion (same one as last picture):


It has been 25 days since inoculation and most of the jars look disgusting. though I am a noob, I've not seen anything that seems to be contamination (edit: at the time i did not know what bacterial contamination looked like). I did notice some yellow and orange spots, but assumed that this was pigmentation from the bee pollen, since the spots have not seemed to change size at all.

it was between 2 and 3 wks after inoculation that they started to turn ugly. the substrate became darker, sank toward the bottom of the jar and developed cavities of air throughout. I tried to eliminate these by hitting the tops of the jars. with time they got worse. some spots of mycelial growth on the jars turned brown and died. there are lots of bits of substrate stuck to the sides of the jars, but there is airspace between the cakes and the glass. this is what the majority of jars look like now:




I have tried turning the jars upside down on a rack to allow for co2 to escape, and that is how they are now. still, nothing seems to be getting better. in the case of one jar (which wasnt growing) an extra .5cc was added as an experiment.. no success. (edit::facepalm:)



what worries me now is that the only 2 cakes that are colonizing seem to be degenerating. WHAT IS HAPPENING TO THESE JARS? I havent noticed anything at all that looks like fungal growth, only a sort of wetness and darkness of the substrate.. kind of looks like its rotting.


Edited by aktar (12/09/14 12:10 PM)


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InvisibleDoc_T
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Re: first grow... off to a bad start [Re: aktar]
    #14038317 - 02/27/11 08:24 PM (12 years, 10 months ago)

Hey man, the tall half pints are notorious for not finishing.
You get better results with the wide mouth ones.

Also, you've got bacterial contamination, most likely from the syringe.


--------------------
You make it all possible. Doesn't it feel good?


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Offlineaktar
nerf herder

Registered: 02/27/11
Posts: 58
Last seen: 8 years, 4 months
Re: first grow... off to a bad start [Re: Doc_T]
    #14038391 - 02/27/11 08:34 PM (12 years, 10 months ago)

Can the jars in the last images be saved? They have extensive mycelium growing in them.

But if thats a contam in the other jars, then its gotten into the two fastest colonizers, and I expect the mycelium will be killed if nothing is done (CAN anything be done?)

the solutions that suggest themselves to me are these:

1 — open the jars, get rid of the filthy substrate, rinse the mycelial growth in distilled water, then break up and use to spawn grain jars, OR

2 — use a syringe filled with distilled water, stick it into my jar (along the glass) and scrape it around on the myc.. then inject a little bit of water and suck it back up. this mycelium-water could be used to inoculate the 2 jars which have not been used yet. granted, if they are contaminated then this might not work, but from the looks of it there is a contam-free area that is accessible to the syringe.

What do you think of these 2 ideas? Any better ones?


Where do you think I went wrong? Unless the room they were noc'd in was filthy with contaminants, I think the syringes were sterilized enough. How long should they be held in flame for sterilization?

EDIT: what sort of contam do you think that is?

ALSO: the jars have smelt of margarine or fake butter.. what are they supposed to smell like? it is a sweet smell, but has started to become more sour or fermented as time passes.


Edited by aktar (02/27/11 09:35 PM)


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InvisibleDoc_T
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Re: first grow... off to a bad start [Re: aktar]
    #14038437 - 02/27/11 08:40 PM (12 years, 10 months ago)

Toss, start over.


--------------------
You make it all possible. Doesn't it feel good?


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Invisiblepenhed
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Re: first grow... off to a bad start [Re: Doc_T]
    #14038861 - 02/27/11 09:31 PM (12 years, 10 months ago)

what doc said


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InvisibleDoc_T
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Re: first grow... off to a bad start [Re: penhed]
    #14039056 - 02/27/11 09:54 PM (12 years, 10 months ago)

Blanket, heating pad, bad. Room temp is where it's at. And the right jars.
And following the tek exactly- modern PF Tek, not some outdated version.

RR shows it in his free vids: http://mushroomvideos.com/ and there's a written version here you can print out for easy reference.


--------------------
You make it all possible. Doesn't it feel good?


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Offlineaktar
nerf herder

Registered: 02/27/11
Posts: 58
Last seen: 8 years, 4 months
Re: first grow... off to a bad start [Re: Doc_T]
    #14039197 - 02/27/11 10:09 PM (12 years, 10 months ago)

Quote:

Doc_T said:
Blanket, heating pad, bad. Room temp is where it's at. And the right jars.




yeah. i had a growing suspicion that the incubation method was not suitable; i only settled for that setup because i didnt have a proper incubator — obviously, that was stupid. i should have just gone out and spent some money.

after about 1.5 wks the jars were moved into a more isolated, dark incubation chamber (though, i still used the heating pad... :crazy: but its winter where im at and i dont think room temp will be warm enough.) later i rearranged the setup so that the jars were not in direct contact with the heat source (above it but not touching. ambient temps have been much better since, with a pretty stable ambient 76-80 degrees)

i will certainly toss the 5 jars with nothing good in them. i had been considering it for a while and by this point i have no hope for them (and i'm sure you know your stuff, too)

BUT, for obvious reasons, i really don't want to toss the 'good jars.' now, i understand that just because i 'dont want to' does not mean that throwing them out isnt a good idea.. but there is at least one jar with contamination only on one side and a large chunk (probably 50% of the jar) of healthy-looking, ropy mycelium.

seeing as i am almost out of spores, i'm thinking it would be a waste to toss them without trying my absolute hardest to make it work. better to get the most out of it, instead of just scrapping it, right..? i will hold off on inoculating with the PB syringe (and the last cc of GT) until i find out whether this will work:

probably tomorrow, i will dump out the cakes, and get those foul jars emptied and cleaned. then i will put on some clean gloves, get some alcohol swabs and a knife, and remove all uncolonized substrate. perhaps i will also remove the parts of the cake closest to the contaminants. once the mycelium has been isolated i will rinse with sterile water, put it into a clean ziploc bag and break it up into fine pieces. then i'll add some fresh, non-contaminated substrate, seal off the bag and incubate.

yes? or am i still being stupid?


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OfflineMicroppose
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Re: first grow... off to a bad start [Re: aktar]
    #14039277 - 02/27/11 10:23 PM (12 years, 10 months ago)

Pasteurized in steam for 45 minutes? I hope you sterilized them and what you said was just a mix of words. Your contamination may have been from the medium itself if not :frown:


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Invisiblebiologys
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Re: first grow... off to a bad start [Re: Microppose]
    #14039294 - 02/27/11 10:25 PM (12 years, 10 months ago)

also, 45 minutes isn't even suitable for PC'n, let alone steam sterilizing

I myself dont do cakes, but i'd recommend sterilizing longerrrr


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OfflinePsilocyentist
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Re: first grow... off to a bad start [Re: biologys]
    #14039362 - 02/27/11 10:34 PM (12 years, 10 months ago)

Sterilize for 90 min, use wide mouth half pints, 4 innoc. points, no incubator (unless your house is SUPER cold i.e. <55, at this point, put them in a cooler with a jar of hot water which is changed occasionally), don't wipe syringes after flaming, and don't turn your jars over.

The biggest things I saw which could be causing contams/stalling is your insufficient sterilization time, turning over of your jars, and pint jars. when you turn it over, it disturbs the dry verm layer and can allow contams through. With tall pint jars, the substrate can compress and suffocate the lower part, stalling growth.


--------------------
I know, I know you probably scream and cry

That your little world won't let you go



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Offlineaktar
nerf herder

Registered: 02/27/11
Posts: 58
Last seen: 8 years, 4 months
Re: first grow... off to a bad start [Re: biologys]
    #14039511 - 02/27/11 11:01 PM (12 years, 10 months ago)

Quote:

biologys said:
45 minutes isn't even suitable for PC'n, let alone steam sterilizing




well, i have read a great number of viewpoints concerning this. some swear by the pc for an hour, others have said 30 mins in a steam bath is enough. in my defense, the 2 jars which have not been opened or touched since pasteurization look perfectly normal, and do not show signs of contaminants.


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InvisibleRahz
Alive Again
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Re: first grow... off to a bad start [Re: Psilocyentist]
    #14039528 - 02/27/11 11:03 PM (12 years, 10 months ago)

Quote:

yes? or am i still being stupid?




Everything else being done well, 45 minute steam pasteurization is a good bet for contams. The problem now is that mycelium has a very poor chance of fighting off contams in uncolonized flour. And if you attempt to save part of the cake, it's very likely that the contams will follow. Not really stupid, but trying to save them is effort that will almost certainly be wasted. Do it if you have to, but I would recommend starting your next grow. As said, you need to boil the jars for 1.5 hours because 45 minutes isn't enough. 45 minutes has been recommended when pressure cooking BRF jars, "PF tek for Simple Minds".


--------------------
rahz

comfort pleasure power love truth awareness peace


"You’re not looking close enough if you can only see yourself in people who look like you." —Ayishat Akanbi


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Offlineafrosheen
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Re: first grow... off to a bad start [Re: Rahz]
    #14039647 - 02/27/11 11:21 PM (12 years, 10 months ago)

I don't think anyone noticed this critical detail in OP's post:
Quote:

the substrate was made with the following (for 9 pint jars):




Pint jars. That's the definition of failure with BRF teks.

Use wide mouth half-pints for the win. You'll get a chance to use those pints later for grains or something else if you so choose.


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OfflineMicroppose
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Re: first grow... off to a bad start [Re: afrosheen]
    #14039674 - 02/27/11 11:25 PM (12 years, 10 months ago)

Quote:

afrosheen said:
You'll get a chance to use those pints later for grains or something else if you so choose.




Like sclerotia, wide mouth pints are great for that.


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Offlinei GrOw StUFF
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Re: first grow... off to a bad start [Re: Microppose]
    #14039969 - 02/28/11 12:24 AM (12 years, 10 months ago)

45 minutes isn't long enough and its called sterilizing, not pasteurizing....

and another important thing is don't flip your jars at all.  Make sure they are always pointed up right so you don't disturb the dry layer of verm, it is the filter.  And don't move them much.....Never use pint jars for BRF tek....always the short fat 1/2 pint jars


--------------------
The mushrooms, which grow only during the season of torrential rains, awaken the forces of creation and produce an experience of spiritual abundance, of an astonishing, inexhaustible constitution of forms that identifies them with fertility and makes them a mediation, a means of communion, of communication between man and the natural world of which they are the metaphysical flesh.


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OfflineCaribou_Lou
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Re: first grow... off to a bad start [Re: i GrOw StUFF]
    #14040504 - 02/28/11 06:46 AM (12 years, 10 months ago)

Use grain spawn. You will never use brf again.


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InvisibleDoc_T
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Re: first grow... off to a bad start [Re: Caribou_Lou]
    #14040515 - 02/28/11 06:51 AM (12 years, 10 months ago)

Quote:

aktar said:
well, i have read a great number of viewpoints concerning this. some swear by the pc for an hour, others have said 30 mins in a steam bath is enough. in my defense, the 2 jars which have not been opened or touched since pasteurization look perfectly normal, and do not show signs of contaminants.




90 mins minimum, 2 hours is ok too.


Quote:

Caribou_Lou said:
Use grain spawn. You will never use brf again.[/quote

Grains are awesome, but you need a pressure cooker and clean inoculant.


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You make it all possible. Doesn't it feel good?


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Offlinejacobensis
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Re: first grow... off to a bad start [Re: Doc_T]
    #14040549 - 02/28/11 07:09 AM (12 years, 10 months ago)

i used the tall half pint jars and had great success. 16 jars done- 16 jars colonized but, i PC`d mine for an hour and follow`d RR`s tek strictly, i will also say i put them in a water incubator at 80` and left them alone. they were exposed to light once 2 Weeks in and thats it. i added nothing to the tek- no bee pollen, nothing. Never try to improve something that comes from the MASTER RR.


--------------------
There are 2 types of people, mycophobes and mycophiles R.Wasson


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OfflineKingMob
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Re: first grow... off to a bad start [Re: afrosheen]
    #14040560 - 02/28/11 07:16 AM (12 years, 10 months ago)

Quote:

afrosheen said:
I don't think anyone noticed this critical detail in OP's post:
Quote:

the substrate was made with the following (for 9 pint jars):




Pint jars. That's the definition of failure with BRF teks.

Use wide mouth half-pints for the win. You'll get a chance to use those pints later for grains or something else if you so choose.




I was gonna say...

Either those are pint jars or this guy/gal has the tiniest baby hands I've aver seen!!!!

We got ourselves a 9 yr old mush grower. :laugh2:

Like the guy in the BK commercial. 
"Guys I would sterilize for longer but...    I just have these tiny hands. Half pints make my hands look even smaller! I don't think I could load them in again."

"Dont worry,  I'll hold your halfpints for ya"

"Really! You'd do that!"  :awejeez:

For real tho, I've used pints before. They hardly ever colonize completely unless you have a fast growing strain.

Not to say they won't work(I have had 3/4 of the car colonize and I've cut off the uncolonized part and used the rest) but it's another step that could cause contamination, which you don't want, especially since you are starting and thus itching for success!

So as most have suggested, get smaller jars or switch to grains. Now if you can't get a hold of widemouth half pint, regular mouth ones will work but will take longer to colonize.

And if you can't get those (I cant get any kind of canning jar where I'm from) then at least sterilize for longer.

Remember that most instructions for sterilization of brf jars with steam refer to the halfpint jars, so if you are using pint jars it will help to leave them longer in the steamer.

I would follow RR's suggestion (seen in his videos) of steaming for 90 mins.

But since you are using pint jars I'd say 2 hrs. This might not be necessary, but it cant do any harm either and could maybe, possibly help. :grin:


Edited by KingMob (02/28/11 07:21 AM)


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OfflineKingMob
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Re: first grow... off to a bad start [Re: jacobensis]
    #14040584 - 02/28/11 07:26 AM (12 years, 10 months ago)

Quote:

jacobensis said:
i used the tall half pint jars and had great success. 16 jars done- 16 jars colonized but, i PC`d mine for an hour and follow`d RR`s tek strictly, i will also say i put them in a water incubator at 80` and left them alone. they were exposed to light once 2 Weeks in and thats it. i added nothing to the tek- no bee pollen, nothing. Never try to improve something that comes from the MASTER RR.




I agree that it would be good to leave it simple and use what works with the least materials. I think that the simpler the procedure and materials, the least problems you will run into.

Once you have had success with the simple recipes and methods, you can add and change as you like.


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Offlineaktar
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Registered: 02/27/11
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Re: first grow... off to a bad start [Re: KingMob]
    #14042894 - 02/28/11 03:20 PM (12 years, 10 months ago)

Quote:

jacobensis said:
i used the tall half pint jars and had great success. 16 jars done- 16 jars colonized but, i PC`d mine for an hour and follow`d RR`s tek strictly, i will also say i put them in a water incubator at 80` and left them alone. they were exposed to light once 2 Weeks in and thats it. i added nothing to the tek- no bee pollen, nothing. Never try to improve something that comes from the MASTER RR.




yes, i suppose i shouldn't have tried to deviate from the tek on my first attempt.. :blush:

Quote:

KingMob said:
So as most have suggested, get smaller jars or switch to grains. Now if you can't get a hold of widemouth half pint, regular mouth ones will work but will take longer to colonize.

And if you can't get those (I cant get any kind of canning jar where I'm from) then at least sterilize for longer.

Remember that most instructions for sterilization of brf jars with steam refer to the halfpint jars, so if you are using pint jars it will help to leave them longer in the steamer.




well, sad to say that i can get half-pint jars; the reason i didnt was because i thought 'bigger cakes, more mushrooms, yeah?'

hehe, i will definitely follow those videos more closely (i mean — perfectly) next time. it's pretty heartbreaking to watch these babies deteriorate :frown: i will keep you all posted on my efforts to salvage the mycelium.


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Offlineafrosheen
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Re: first grow... off to a bad start [Re: aktar]
    #14044303 - 02/28/11 06:45 PM (12 years, 10 months ago)

Quote:

it's pretty heartbreaking to watch these babies deteriorate




Trust me, we've all been there brother. Losing a cake sucks, but losing a monotub to a giant spot of trich before the first flush is a heartbreaker.


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


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OfflineKingMob
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Re: first grow... off to a bad start [Re: afrosheen]
    #14047028 - 03/01/11 05:45 AM (12 years, 10 months ago)

Quote:

afrosheen said:
Quote:

it's pretty heartbreaking to watch these babies deteriorate




Trust me, we've all been there brother. Losing a cake sucks, but losing a monotub to a giant spot of trich before the first flush is a heartbreaker.



:whathesaid:


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InvisibleSillicybin
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Re: first grow... off to a bad start [Re: i GrOw StUFF]
    #14047263 - 03/01/11 07:39 AM (12 years, 10 months ago)

Quote:

i GrOw StUFF said:
and another important thing is don't flip your jars at all.  Make sure they are always pointed up right so you don't disturb the dry layer of verm, it is the filter.



QFT.  You absolutely need to keep jars without a filter upright.  The 'flipping to drain CO2' advice is bunk.  You're just inviting contams to the party.  Even with a filter, I try to keep uncolonized substrate off the lid to help prevent wicking contams off the filter.

Especially on PF style cakes, you want to disturb the jars as little as possible, while being as clean and sterile as possible when you are near them.


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