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OfflineLeastham
Mycologist
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Registered: 06/15/09
Posts: 126
Last seen: 6 years, 3 months
Trich... Trich... Trich Again... Please Help
    #23428970 - 07/10/16 10:46 AM (7 years, 6 months ago)

Alright so here is the deal.
ill number the attempts

1. GREAT Success (Spores to Grain 2 Grain)
had done a PE Bulk Grow a while back and had great success 12 tubs no issues.

2. SUPER FAIL
now i had tried the same process, with 6 tubs from some LC from the first run it looked healthy in WBS jar, they finished colonizing and then i mixed the coir and verm and used my steam pasteurizer that i used the first time...
All ended up getting Trich... So i tossed them.

at first i thought it was the LC so i got some NEW APE spores

3. FAIL
made some new jars and grain 2 grain to have a bunch more, they all colonized and before i rush in to making 6 tubs that fail again i go and get a NEW mini tub drill the holes taped them up and used the same coir i had made before since it was an extra and never was opened after... so i pasteurized it again. mixed 1 quart of spawn with about the same amount of sub, sat in dark place for about 10 days and it was fully colonized... about 3-4 days after i get Trich.

Pretty frustrated now, i think back on what i could have done wrong this time, knowing almost immediately that the old coir was probably the issue, and maybe i didnt run my pasteurizer correctly, as the jars look healthy as can be.

so i do some forum searching find some teks for coir and pasteurizing.

4. Fail
this time... i mix water, coir, and verm in 2 small 2mil bags i got from uline that someone suggested works great added a square of ductape stabbed it through the center with a meat therm that was washed and cleaned with alcohol(it was new), used my pc pot stuck them in put a plate over the top(as suggested in the TEK) put it on medium till the internal temp got to 135 then killed the heat and let it sit till it was cool, used the same tub(i was told it would be fine) i scrubbed with soap and let soak in a ridiculously concentrated bleach/water bath for hours then rinsed and dried and taped up

i shower, them get my 1 quart spawn birth it into a freezer gallon bag, break it up. put latex gloves on, rip open the sub then mix the two together, 1 quart spawn, 1 quart sub, put in a dark place 72-74 degrees... Colonizes super fast and looks great then 10 days later, i add polyfil loose on top holes and a bit more stuffed on the bottom holes... Turn the 12 on 12 off in a 72-74 Degree room,

3 days in it shows signs of trich and i spot the trich YET AGAIN.

I am not to happy with myself, as i had done twelve 64 quart tubs prior easily and without a problem. flushed 3-4 times each... and have done multiple others that worked Great.

.:NOW:.
i will do a 5th run these are my thoughts.
1. Try the Bucket Style of Pasteurizing (using this Tek)
other than that my pasteurization process i cant figure out what could possibly be the issue.

???Questions???
1. What are the chances its the tub after a super scrub down and a bleach bath? Because it did fail before the tub had trich in it when it was new.

2. Is it possible my coir was not mixed right the first couple of times? maybe more Verm?

3. Could my Mixture Be to Wet? I usually Squeeze it until nothing comes out

PLEASE HELP ME :frown:





Edited by Leastham (07/10/16 10:56 AM)


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OfflineHybridprX
Biodegrader of coir
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Registered: 01/29/08
Posts: 2,588
Loc: Canada Flag
Last seen: 6 years, 4 months
Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23428982 - 07/10/16 10:53 AM (7 years, 6 months ago)

It's your spawn op

Your syringe must have been a bit dirty....you'll need to isolate out a few healthy sectors on agar and test them before doing another G2g.


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


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OfflineLeastham
Mycologist
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Registered: 06/15/09
Posts: 126
Last seen: 6 years, 3 months
Re: Trich... Trich... Trich Again... Please Help [Re: HybridprX]
    #23429002 - 07/10/16 10:58 AM (7 years, 6 months ago)

Quote:

HybridprX said:
Your syringe must have been a bit dirty....you'll need to isolate out a few healthy sectors on agar and test them before doing another G2g.




even with a NEW Spore Syringe? from a vendor?


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Invisibleelasticaltiger
Like Tigers in Coitus
Male


Folding@home Statistics
Registered: 06/24/13
Posts: 8,070
Trusted Cultivator
Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23429536 - 07/10/16 02:44 PM (7 years, 6 months ago)

How are you inoculating your jars?

Do you use a SHIP or do you open the jar and just squirt the spores in?


--------------------
First time growing cakes? DON'T make a Shotgun Fruiting Chamber

The Shmuvbox. - The Old TC's Like it :shrug:

Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger!
No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit.

"The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT

Pastywhite's Easy Agar Tek (PastyPlates)

Tiger Drop Video Demos By munchauzen

Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson

EZEKIEL 23:20


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OfflineHybridprX
Biodegrader of coir
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Registered: 01/29/08
Posts: 2,588
Loc: Canada Flag
Last seen: 6 years, 4 months
Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23429569 - 07/10/16 02:57 PM (7 years, 6 months ago)

Quote:

Leastham said:
Quote:

HybridprX said:
Your syringe must have been a bit dirty....you'll need to isolate out a few healthy sectors on agar and test them before doing another G2g.



even with a NEW Spore Syringe? from a vendor?




It's not common for vendor syringes to be contaminated but yes...could have been interduces by accident t by the vendor or it could have accidently cross contaminated at some point...

Agar is your best hope.


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


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OfflineTheDeathryderIsBac
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Registered: 12/25/15
Posts: 319
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Last seen: 3 years, 6 months
Re: Trich... Trich... Trich Again... Please Help [Re: HybridprX]
    #23429581 - 07/10/16 03:01 PM (7 years, 6 months ago)

Definately the syringe.  Keep in mind that there is no such thing as a sterile print, some contaminate spores will always be present.  This is why spores to grain/LC is generally discouraged.  Agar will save you a LOT of time and headaches.


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Invisibleelasticaltiger
Like Tigers in Coitus
Male


Folding@home Statistics
Registered: 06/24/13
Posts: 8,070
Trusted Cultivator
Re: Trich... Trich... Trich Again... Please Help [Re: TheDeathryderIsBac]
    #23429594 - 07/10/16 03:06 PM (7 years, 6 months ago)

The syringe may have been clean but your technique might be contaminating your work.

Are you using an SAB or are you doing open air?


--------------------
First time growing cakes? DON'T make a Shotgun Fruiting Chamber

The Shmuvbox. - The Old TC's Like it :shrug:

Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger!
No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit.

"The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT

Pastywhite's Easy Agar Tek (PastyPlates)

Tiger Drop Video Demos By munchauzen

Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson

EZEKIEL 23:20


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OfflineLeastham
Mycologist
Male


Registered: 06/15/09
Posts: 126
Last seen: 6 years, 3 months
Re: Trich... Trich... Trich Again... Please Help [Re: elasticaltiger]
    #23430113 - 07/10/16 06:58 PM (7 years, 6 months ago)

Quote:

elasticaltiger said:
Are you using an SAB or are you doing open air?




i use self sealing lids, made from silicone, with tyvek on both sides of the hole on the lid. i will be trying again my LC looks healthy and nice, and i got a few more spore syringes, also got some agar going from the original clone of the first 12 tubs, i just transferred 3 slices into some nice trays.

i dont understand how it could go so well, then with different jars and different ways of inoculation i can get the same issue everytime.


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InvisibleMad Season
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Registered: 09/16/12
Posts: 12,666
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Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23430127 - 07/10/16 07:07 PM (7 years, 6 months ago)

Quote:

Leastham said:
i dont understand how it could go so well, then with different jars and different ways of inoculation i can get the same issue everytime.



As said by every person who got into this in the history of this hobby.

IDK any vets who didn't start off doing open air g2gs successfully, and now they don't even bother with slightly bacterial jars.

Consider this the moment when you have to be diligent. Now that you have a huge spore load, you HAVE TO HAVE clean spawn. Not just like half clean spawn. ALWAYS start every project on agar, spores and clones. DEFINITELY do all inoculations in a SAB, PC grains for a minimum of 2 hours, NEVER trust vendor syringes, ONLY trust clean cultures on agar, ALWAYS test LC cleanliness on agar, get some better filters too.. Tyvek is shite and the pores get bigger after a couple PC cycles, and please for the love of god read up on sterile procedures. I got some of my agar procedures in the improve your sterile techniques link in my sig. Munch has some epic videos, bod(TC) has a great agar resource too.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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InvisibleSupalemonhaze
Spore syringe hater.
Male


Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
Re: Trich... Trich... Trich Again... Please Help [Re: Mad Season]
    #23430187 - 07/10/16 07:28 PM (7 years, 6 months ago)

If that LC was made with spores, it's no wonder you saw green. I wouldn't even g2g a spore inoculated jar, you're already starting out with an unreliable inoculant, last thing you want is the bacteria that accumulates during g2gs. Not to mention that 1 slip up with your master jar and every recieving jar gets fucked.

Agar wedges are your friends. There is nothing else I would rather use. The folks who complain about the time it takes for wedges to colonize a grain jar have no idea on how to rotate your tubs and how to inoculate batches of grains routinely. My contam rate has never been lower ever since I started doing just that.


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OfflineLeastham
Mycologist
Male


Registered: 06/15/09
Posts: 126
Last seen: 6 years, 3 months
Re: Trich... Trich... Trich Again... Please Help [Re: Supalemonhaze]
    #23430554 - 07/10/16 09:18 PM (7 years, 6 months ago)

Quote:

Mad Season said:
IDK any vets who didn't start off doing open air g2gs successfully, and now they don't even bother with slightly bacterial jars.

Consider this the moment when you have to be diligent. Now that you have a huge spore load, you HAVE TO HAVE clean spawn. Not just like half clean spawn. ALWAYS start every project on agar, spores and clones. DEFINITELY do all inoculations in a SAB, PC grains for a minimum of 2 hours, NEVER trust vendor syringes, ONLY trust clean cultures on agar, ALWAYS test LC cleanliness on agar, get some better filters too.. Tyvek is shite and the pores get bigger after a couple PC cycles, and please for the love of god read up on sterile procedures. I got some of my agar procedures in the improve your sterile techniques link in my sig. Munch has some epic videos, bod(TC) has a great agar resource too.




i use a glove box with alot of oust.


Quote:

Supalemonhaze said:
If that LC was made with spores, it's no wonder you saw green. I wouldn't even g2g a spore inoculated jar, you're already starting out with an unreliable inoculant, last thing you want is the bacteria that accumulates during g2gs. Not to mention that 1 slip up with your master jar and every recieving jar gets fucked.

Agar wedges are your friends. There is nothing else I would rather use. The folks who complain about the time it takes for wedges to colonize a grain jar have no idea on how to rotate your tubs and how to inoculate batches of grains routinely. My contam rate has never been lower ever since I started doing just that.




i cloned one of the nicest specimens from my previous grow to make the lc.


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InvisibleSupalemonhaze
Spore syringe hater.
Male


Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23430560 - 07/10/16 09:20 PM (7 years, 6 months ago)

Cloned to agar?


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InvisibleMad Season
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Registered: 09/16/12
Posts: 12,666
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Re: Trich... Trich... Trich Again... Please Help [Re: Supalemonhaze]
    #23430571 - 07/10/16 09:23 PM (7 years, 6 months ago)

SAB > GB hands down. Oust is like super expensive compared to water and soap. Is it flammable? (I really don't use it) How do you even flame shit with a glove box?


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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InvisibleSupalemonhaze
Spore syringe hater.
Male


Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
Re: Trich... Trich... Trich Again... Please Help [Re: Mad Season]
    #23430589 - 07/10/16 09:32 PM (7 years, 6 months ago)

I think every aerosol is flammable. Oust is kind of like foam though, it won't saturate a box with flammable gas, it settles pretty quickly.

I agree though, good sterile technique is all you need. Depending on something like oust will not get you far.


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OfflineLeastham
Mycologist
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Registered: 06/15/09
Posts: 126
Last seen: 6 years, 3 months
Re: Trich... Trich... Trich Again... Please Help [Re: Mad Season]
    #23430617 - 07/10/16 09:45 PM (7 years, 6 months ago)

Quote:

Supalemonhaze said:
Cloned to agar?




yes

Quote:

Mad Season said:
SAB > GB hands down. Oust is like super expensive compared to water and soap. Is it flammable? (I really don't use it) How do you even flame shit with a glove box?




curious to what you mean by SAB


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InvisibleSupalemonhaze
Spore syringe hater.
Male


Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23430618 - 07/10/16 09:47 PM (7 years, 6 months ago)

Still air box. Basically a glove box without the gloves. It's far better at keeping the air still, which is the only reason why folks use GBs and SABs.


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InvisibleMad Season
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Registered: 09/16/12
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Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23430626 - 07/10/16 09:50 PM (7 years, 6 months ago)

Quote:


This is a picture of my 110 qt clear still air box:

Notice it's wet/condensated inside. That's what it's supposed to look like the whole time you're working with it.


I took the cut outs of the holes and taped them to make little doors. This is to keep the air still if I'm waiting for things to cool, or if I want a smoke break.

My SAB holes are 6 inches. Honestly this is the MINIMUM diameter. I took incense and lit it in my box (kudos to kizzle for the idea) and found the arm holes moved the most air (smoke) when moving my arms around, and in and out. By increasing the size of the arm holes, and working in a fairly still room with the vents turned off .. I've been finding that moving your arms in and out will cause no air movement. And thus very rare contams




Also you flame outside the box, keeping the air inside the box still.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


Edited by Mad Season (07/11/16 01:33 PM)


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OfflineLeastham
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Re: Trich... Trich... Trich Again... Please Help [Re: Mad Season]
    #23430651 - 07/10/16 10:05 PM (7 years, 6 months ago)

almost the same setup i ditched the gloves of my GB. so if my spore syringes are bad then can i get a hint to a non contaminated vendor?


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InvisibleSupalemonhaze
Spore syringe hater.
Male


Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
Re: Trich... Trich... Trich Again... Please Help [Re: Leastham]
    #23430665 - 07/10/16 10:09 PM (7 years, 6 months ago)

There is no such thing as a sterile syringe. Agar is the way to go if you want the best chance of getting clean spawn.


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OfflineLeastham
Mycologist
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Registered: 06/15/09
Posts: 126
Last seen: 6 years, 3 months
Re: Trich... Trich... Trich Again... Please Help [Re: Supalemonhaze]
    #23430687 - 07/10/16 10:20 PM (7 years, 6 months ago)

alright so i got some agar dishes going... how you know if its clean or not it looks white and good.

i used pasties tek with the mini rounds but i can see through them.


Edited by Leastham (07/10/16 10:27 PM)


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