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SmokinErb
Stranger
Registered: 01/21/11
Posts: 193
Last seen: 11 years, 10 months
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Help identifying the source of contamination needed.
#16798934 - 09/06/12 02:34 PM (12 years, 4 months ago) |
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I've ran into a recent bout of trich contamination prior to the first flush. Quick run down of my methods as follows:
Grain jars (milo) are PC'd for 60-90 minutes at 15psi and transferred to a semi-sterile laboratory with HEPA filtered intake and airlock chamber.
inoculation methods varied - some were injected via silicon port, others were open air.
Fruited 3 MS monotubs with good success, cloned a fruit into LC.
Contam rates for grain jars since taking clone have been 1 contam for 60 jars (trich), contamed jar was promptly removed from lab and lab was disinfected.
Substrate is coir and verm, pasteurized in a hot water bath - coir/verm is placed in a pillow case and then placed in a cooler, water is heated to 170f and cooler is filled. A few quart jars are placed on top of the substrate to hold it under the water level for 90+ minutes.
Monotubs are 12 gallon bins, I use 4 quart jars of spawn per monotub and approx 1 1/3 brick of coir. Monotubs colonize quickly, surface is 80% colonized in under a week before I notice the green spots. I do not prepare monotubs in the laboratory as they are not disinfected but merely pasteurized, I mix them on the living room table using standard clean procedures.
100% contam rate in my last 3 monotubs, all prior to 1st flush. I did attempt to case one monotub, casing contaminated, but caught it early and removed the casing. Attempting to salvage this o.e as only a small spot of the sub contaminated and it was cut out and pins are developing. Any tiny spotz of trich are checked for and removed several times daily (only appeared once since initial discovery)
This is not ideal, however I am moving out in 2 weeks and am just trying to salvage one flush out of this monotub to have some kibd of reward for my work. Not the source of contamination anyway as all 3 monotubs were done within 48 hours of each other and are isolated during incubation.
I have always had mixed results with cubes, which is why i built a lab for sterile work. Should I be mixing my coir/spawn in the lab as well? My only other suspicion is possibly a faulty thermometer used to measure the temp of pasteurization water.
Also no ph was ever adjusted as i lost my ph test kit. Possible cause? I believe i should be hydrating/pasteurizing with water that is roughly a ph of 8?
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Johnny Quest
Mycology Lover



Registered: 08/31/08
Posts: 321
Loc: Just moved to the British...
Last seen: 6 years, 6 days
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Re: Help identifying the source of contamination needed. [Re: SmokinErb]
#16799147 - 09/06/12 03:18 PM (12 years, 4 months ago) |
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Perhaps your substrate needs to be cooked differently. A good way to go about it is to bring the sub to field capacity and then heat it on your stove to 140-160 degrees and hold it there for an hour. Otherwise you cant be sure pockets within the sub were left uncooked.
Another possibility is that your grain jars were a little too wet. If too much water is in your jars, parts of your grain won't colonize in the safety of the jar so when you mix it with coir, the grain can support contam growth.
I wish you luck on your current grow...my last monotub had a small contam but I was able to get 4 oz out first! You've got hope.
-------------------- Studying mycology since: August, 2008
A typical monotub grow of a multi-spore rye spawn with some Golden Teachers: inoculated on day one and saw first signs of growth on day 5. On day 17 jars were 15% - 30% colonized so shook em up. On day 25 I mixed the spawn and sub in the monotub and on day 50 harvested the first flush. Just under 4 ounces dry in 1 month 19 days!
One Love!
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SmokinErb
Stranger
Registered: 01/21/11
Posts: 193
Last seen: 11 years, 10 months
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Re: Help identifying the source of contamination needed. [Re: Johnny Quest]
#16799432 - 09/06/12 04:01 PM (12 years, 4 months ago) |
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I've harvested contaminated monos before, I just take em to another location 
A major reason I use the hot water bath method is because I usually pasteurize 4 bricks of coir at a time, and also, wouldnt the coir in contact with the bottom of the pan be effectively sterilized? I suppose I could do frequent smaller batches, but this method has worked for me in the past, so I figured why change it? What if i extended the pasteurization time to say 2 hours? i forgot to mention that I break the coir bricks up into small pieces to ensure they reach field capacity and tempalmost immediately.
I do occasionally get jars a little too wet and they'll have spots not colonize, but those get thrown out. I did use these jars as soon as they appeared fully colonized, perhaps they had spots in the middle that hadn't quite yet been colonized. I've got more jars ready to go, I'll let them incubate a few more days longer to be sure.
Frusturating for sure. I've never had any contaminate other than trich in this past year, built a lab to try to reduce contam rates and so far, they've skyrocketed. I think I'll stick with my water bath method for now, but i'll hydrate the coir prior to pasteurizing and see if that helps. Building a bulk steam pasteurizer is next on my list when I get the funds to do so anyway.
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Johnny Quest
Mycology Lover



Registered: 08/31/08
Posts: 321
Loc: Just moved to the British...
Last seen: 6 years, 6 days
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Re: Help identifying the source of contamination needed. [Re: SmokinErb]
#16805593 - 09/07/12 02:47 PM (12 years, 4 months ago) |
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Oh yes... I didnt realize that you didn't presoak your grains...some folks say the presoak causes trich endospores to germinate thus allowing you to zap them in the pressure cooker. Otherwise the ungerminated endospores can survive the pressure cooker and make their move when the mycelium is weak (like right as you spawn to bulk)....Ive had good results without the presoak but ever since Ive heard about it, Ive done it....I dont know how true all that info is but it sounds reasonable to me.
-------------------- Studying mycology since: August, 2008
A typical monotub grow of a multi-spore rye spawn with some Golden Teachers: inoculated on day one and saw first signs of growth on day 5. On day 17 jars were 15% - 30% colonized so shook em up. On day 25 I mixed the spawn and sub in the monotub and on day 50 harvested the first flush. Just under 4 ounces dry in 1 month 19 days!
One Love!
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HybridprX
Biodegrader of coir



Registered: 01/29/08
Posts: 2,588
Loc: Canada
Last seen: 7 years, 4 months
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Re: Help identifying the source of contamination needed. [Re: Johnny Quest]
#16805662 - 09/07/12 02:58 PM (12 years, 4 months ago) |
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Soaking for 24 hours causes the endospores to germinate, they're bacteria encased in a hard shell capsule called a capsid, its bacterias way of preserving itself for decades. If you submerge them in water you have initiated a environment favorable to their germination and also hydrate the grains at the same time, soaking is a good wise thing to ensure success.
Trich is a mold that floats freely everywhere we go, besides a 100% controlled laboratory with a pressurized air lock system which home cultivators do not have....unless you're RR maybe...
Do you have alot of carpeting in your living room....heavily fabricated rooms will doom mycelium because of the amount of trich present in the air. Fabric holds trich like hydrogen bonds to oxygen.
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SmokinErb
Stranger
Registered: 01/21/11
Posts: 193
Last seen: 11 years, 10 months
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Re: Help identifying the source of contamination needed. [Re: HybridprX]
#16816841 - 09/09/12 11:50 AM (12 years, 4 months ago) |
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I actually do have a pretty good laboratory with an airlock chamber, I just didn't mix the spawn and sub in there because I was concerned that the pasteurized sub shouldnt go into the lab at all since it isnt sterile? I can always mix it in there in the future, i just make a habit of taking nothing that hasnt been disinfected in the lab.
Grains are soaked for 24 hours, didn't mention that. I did just shake my 90% colonized jars, hopefully to distribute any existing mold spores around, allowing them to show themselves before using the jars in the monotub. My logic is that if there is a small contaminated spot in the middle, it will get distributed in the grains which would then spread quickly, eliminating the possibikity of using contaminated grain at the cost of a few extra days waiting.
I'm chalking it up to pasteurization. It may be possible that some of the coir was over the water level in the cooler.
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NamRedruM0016
***THE NOOB***



Registered: 10/17/10
Posts: 460
Last seen: 5 years, 10 months
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Re: Help identifying the source of contamination needed. [Re: SmokinErb]
#16818852 - 09/09/12 06:24 PM (12 years, 4 months ago) |
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that is why i always put my bulk mix into a pillow case and make sure it is well saturated and submerged throughout the entire pasteurization procedure...
-------------------- "Lack of curiosity bored the Cat to death"
MY TRADE LIST:
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