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Surfer77
Stranger

Registered: 03/27/14
Posts: 2
Last seen: 9 years, 11 months
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Epic Failure - Holly Contamination - Please Help
#20263135 - 07/12/14 02:21 PM (10 years, 6 months ago) |
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Please provide some guidance. I’ve done my homework and read hours upon hours forum posts but I’m still failing miserably after 6-8 attempts. I keep getting a greyish mold in my monotubs right before/when the tubs are ready to fruit and it destroys everything. I’ll layout my process step by step but I’m following the teks I’ve read on this site. Sorry for the long post but I want to provide as much detail as possible.
Preparation: As my substrate I’m using birdseed that I bought from the grocery store. I soak the seed in a bucket of water overnight then rinse in a kitchen screener until the water is clear. I then put the seed in a pot and steep on the stove for 30 minutes. I do not let the water boil but get it hot enough that heavy steam is raising from the water. Next I rinse the seed in a screener until clear water is running through it and let it drain in a colander so the exterior can dry (approx. 1 hour) I then load 7 wide mouth quart jars half way with the prepped seed. I stuff poly fill in the hole of the lid pretty tight, cover with foil and pressure cook the seed for 1.5 hours. I then let the cooker lose pressure, drain the water, shake the jars and put them back in the cooker with the lid locked overnight while the cool.
Inoculation: I originally use spore syringes to inoculate my WBS. I don’t have a hood so I use the half bath in my house. Prior to starting inoculation I remove everything from the bathroom, spray copious amounts of Lysol, and whip all surfaces with bleach cleaner, close pour bleach in the toilet, close all vents, pour bleach down the sink drain and close the drain. I then place all my materials in the bathroom along with a Hepa/Ionic air purifier and let it run for at least an hour. I change into clean long sleeve shirt, long paints, and a hat. Once I enter the bathroom I put on rubber gloves and a face mask and wipe all the flat surfaces I will be working on/near with alcohol as well as all the items I’ve placed in the bathroom. Once I’ve cleaned everything I turn off the air purifier, open the pressure cooker, remove the foil and wipe the jars with alcohol and do the same to the spore syringe. I keep a paper towel saturated in alcohol close while I get the tip of the needle red hot then immediately put the needle on the paper towel and fold the towel over the needle and squirt out a small bit of spore solution. I then pick up the syringe while the needle is still covered with the paper towel and place it by the polyfill in the jar lid. I add a bit more than 1cc to the jar and pull out the syringe immediately covering it again with the alcohol paper towel. I repeat 6 more times. Once I’ve inoculated the jars I pour alcohol on coffee filters and cover each jar with a filter and wrap with a rubber band. I then spray more Lysol into a cardboard box and place the jars in the box and close the box and store until the jar is fully colonized with mycelium.
I’ve also used the same process for G2G innoculations once the jars are fully colonized minus the syringe.
Storage: I’ve removed everything out of the closet in my office, taped/stapled thick plastic over the carpet and wiped the walls and shelves with bleach water or alcohol, spray Lysol and put the box of jars in on the shelf. The closet is approximately 10x3 with two sliding doors. I’ve made a gasket that overs the vertical gaps in the doors except where they meet to open. There are still openings at the top and bottom of the doors but I keep the hepa/ionic air purifier in the room on medium. The purifier is rated for 300 square feet so it should be plenty for a small closet. This is also where I keep my monotubs. I work in my office everyday where the closet is located but do not open the doors unless necessary to tend to the operation. I have two cats who walk into the office but they do not get in the closet. Each time I enter the closet I spray Lysol before and after entering. I have plants in my house but not in the office. Unfortunately I don’t have a better area to store everything other than my office closet but I make every effort to keep it sterile. I’m border line OCD so my house always stays very clean.
Mycelium Growth: I have had no signs of contamination with any of the WBS jars I’ve inoculated. They are all bright white and smell like fresh shrooms.
Bulk Substrate: Originally I was using wheat straw as my bulk substrate. I have a friend that has had great success with straw and I’m a huge fan of his end products so I wanted to mirror his process. I bought straw from a craft store to use as my substrate. He was mentoring me but had to travel out of the country for work and I have not been able to communicate with him on a regular basis. I’ve spawned six tubes with wheat straw and each time they get a grey mold on them as soon as they are fully colonized an start to pin. I’ve been able to yield some shrooms twice but the mold takes over the tub before it can grow a full yield. After six failures I stopped my operation I’ve switched to 50/50 coir verm using Damon’s 50/50 tek. I read on the site that straw is very susceptible to contamination so I’ve switched to the coir verm to see if that was the problem but my first tub has become contaminated right before it was completely colonized. It has taken close to 20 days to reach full colonization and at that time the grey mold appeared.
Spawning the Substrate: I’ll skip my wheat straw spawning process because I plan on staying with the 50/50 tek moving forward. After my failures with the straw I shut down the operation and did a serious scrub on the entire office and closet. I wiped down every single item in the office and walls/floors of the empty closet with a strong bleach water solution. I then started the process over again with new spores.Once my WBS jars are fully colonized I place prep my coir verm following Damon’s 50/50 tek. I thoroughly clean my bucket with bleach water and let it dry. Prior to starting the process I take the same sterilization steps in the half bath listed above. I Place a block of coir in the bucket along with 2 quarts of verm. I then boil. 4-4.25 quarts of water and pour it into the bucket and close the lid. I put the bucket in the bathroom and keep the air purifier running. After 30 minutes to an hour I open the bucket and stir with a sterilized grill spatula with a long handle. After the coir/verm has cooled I wipe down my tub with bleach water. I’ve been using the same tubs but clean the ever living shit out of them. I also wipe the entire tub with alcohol immediately before I open the bucket while wearing rubber gloves and a face mask. Once I have my jars in bathroom and they’ve been wiped down with alcohol I open the bucket and pour the majority of the coir/verm mixture into the tub and mix 6 quarts of fully colonized WBS jars (all smell like shrooms) and cover the mixture with the remaining coir/verm approx. ¼ inch deep. All the holes on the tub are taped shut except for the top two holes on the long sides of the tub and those are stuffed tightly with polyfill. I use the clear plastic window insulation to seal the top of the tub because plastic wrap was proving to be a pain in the ass. The window insulation is clear and seals nicely once I press it down and run the hair drier over it quickly. I then put the lid on the tub and place it in the same office closet. I set it on top of a plastic container that’s been wiped down with bleach water to keep it off the ground and didn’t touch it for 10 days. After 10 day I’ve looked at it and when it was starting to get close to full colonization I took off the lid (white) and turned on my 6,500k fluorescent light on a 12/12 timer. I looked at it a couple of days ago and though I saw contamination but wasn’t sure so I gave it a few more days. The pictures are what it looks like now. Obviously not what I’m aiming for.
I can’t figure out what I’m doing wrong but I’m starting to get really frustrated and could really use some guidance. Any advice you can offer would be greatly appreciated.



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Sanguin3
Optimist

Registered: 10/19/13
Posts: 2,273
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Re: Epic Failure - Holly Contamination - Please Help *DELETED* [Re: Surfer77]
#20264157 - 07/12/14 07:18 PM (10 years, 6 months ago) |
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Post deleted by Sanguin3
Reason for deletion: .
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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,870
Last seen: 2 months, 8 days
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Re: Epic Failure - Holly Contamination - Please Help [Re: Sanguin3]
#20264385 - 07/12/14 08:13 PM (10 years, 6 months ago) |
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Quote:
I’ve also used the same process for G2G innoculations once the jars are fully colonized minus the syringe.
This is a problem. You may be able to get away with syringe inoculations without a SAB but not g2gs.
Also alcohol doesn't kill mold spores, don't touch the syringe needle with anything after it's been flame sterilized or you risk contaminating it again.
The coffee filters over jars don't help anything either.
Oh and remember that just because a jar looks clean doesn't mean it is. That's Trichoderma and it's notorious for growing in spawn jars but not showing itself until after it's been spawned.
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Surfer77
Stranger

Registered: 03/27/14
Posts: 2
Last seen: 9 years, 11 months
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Re: Epic Failure - Holly Contamination - Please Help [Re: Kizzle]
#20269736 - 07/14/14 06:03 AM (10 years, 6 months ago) |
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Thanks guys,
I plan on making a still air box to work in from now on. Any additional suggestions on inoculation process that I should change? Do you think that the contamination is taking place during the inoculation process or when I spawn the bulk substrate? I have jars ready to be spawned into bulk but they are a G2G from the jars that contaminated in the pictures above should I throw them out and start over from a new syringe?
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TheEaglesGift
The Nagual


Registered: 04/10/11
Posts: 10,554
Loc: Ixtlan, Mexico
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Re: Epic Failure - Holly Contamination - Please Help [Re: Surfer77]
#20272097 - 07/14/14 04:44 PM (10 years, 6 months ago) |
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If you did your grain to grain transfers in open air, throw that spawn out. Don't waste your time on it. You really need a SAB for g2g.
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Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,870
Last seen: 2 months, 8 days
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Re: Epic Failure - Holly Contamination - Please Help [Re: TheEaglesGift]
#20272180 - 07/14/14 04:58 PM (10 years, 6 months ago) |
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During the inoculation definitely with that amount of mold. Always smell your jars before you spawn and do it after you shake the grains apart. Trich can have a coconut odor or a musty odor and with species no odor at all so be careful.
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Donsanpedro
Stranger

Registered: 07/05/14
Posts: 268
Last seen: 10 years, 5 months
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Re: Epic Failure - Holly Contamination - Please Help [Re: Kizzle]
#20272191 - 07/14/14 05:00 PM (10 years, 6 months ago) |
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Rubbing alcohol does not kill mold spores. Alcohol is better as a germicidal. Also, the CDC has this to say "Uses. Alcohols are not recommended for sterilizing medical and surgical materials principally because they lack sporicidal action and they cannot penetrate protein-rich materials. "
from http://www.cdc.gov/hicpac/disinfection_sterilization/6_0disinfection.html
Its a good read, i highly suggest it.
DSP
Live long and cultivate
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