Paradox
©
Fisana

Jump to content


Photo
- - - - -

Blue Meanies on Rye


  • Please log in to reply
18 replies to this topic

#1 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 04 April 2008 - 06:34 AM

6 days ago I knocked up a jar of organic rye berries from a local health food supplier. This is my first attempt with rye.
I began with a 24 hour soak in water, rinsed and cleaned afterwards (perhaps to much?)
I then boiled the rye for some 20 minutes (to much?) as I got caught up on the phone and left it boiling away.
After this I drained the rye and let the excess evaporate for a while before placing into the jar with a silicone injection port and polyfill FAE on the lid. At this point I was supicious as to wether or not I had let the berries get to soft...err cooked. They aren't mushy or soggy but are quite soft to the touch and squish very easily between my fingers.
I then placed it into the PC for 60 minutes at 13 psi (the maximum of my pc). Upon completion IMHO the berries appeared too bulging and many had popped.
Upon cooling the following morning I innoculated the rye with 2ml of spores from a flame sterilized syringe and mixed up the rye inside a bit by rolling the jar.
Four days later there was already the first signs of mycelia growth but on the fifth day there was a clear contamination from Trichoderma.
The first spot appeared just below the injection port on the surface so I began to speculate that perhaps the port had not sealed properly and thus allowed the contam to enter and fall directly to the surface below the port. However, on day six I could see green spots appearing in three other locations on the sides of the jar as well and well isolated from the original location. Upon emptying the jar the contam was also beginning to show inside the rye also, not just on the sides and top...now, I am doubtful as to the cause of this contamination.
1. The injection port or polyfill filter were not properly sealed?
2. The syringe was contaminated?
3. Not sterilized thoroughly and cooked to much?
The attached photos are after emptying the jar into a tray for further inspection...
Any feedback will be most appreciated!
Thanks

Attached Thumbnails

  • DSC00005.JPG
  • DSC00001.JPG


#2 onediadem

onediadem

    Insidious Drivel

  • OG VIP
  • 16,392 posts

Awards Bar:

Posted 04 April 2008 - 06:53 AM

Actually, your grains do not look as tho they were overcooked at all. Ive cooked them much more and had a 1/4 explode with no issues. As long as they are sterile, that isn't a big problem. After cooking, and then pc'n, I would have to say your contam issue came from your syringe solution. All I can suggest is doing up a pf style jar, or a brf paste and test the syringe before wasting any more grain.

#3 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 04 April 2008 - 07:22 AM

After cooking, and then pc'n, I would have to say your contam issue came from your syringe solution. All I can suggest is doing up a pf style jar, or a brf paste and test the syringe before wasting any more grain.


Thanks for the feedback.
My only doubt to this end is that the same syringe was used to innoculate LC's also and these seem to be doing fine (see attached link)
http://mycotopia.net...-lc-s-cool.html
I believe you've already posted in this thread as well.

#4 onediadem

onediadem

    Insidious Drivel

  • OG VIP
  • 16,392 posts

Awards Bar:

Posted 04 April 2008 - 08:16 AM

Bummer... at least it was just one jar and not twenty...:dead:
If it did come from your lc then other possible contamination points are added like the spore syringe or by its injection. Trich can sporulate in an lc. I have seen it happen years ago and Lazlo was able to take a pic of it happening.
You could let the LC sit undisturbed for a few days and if you see green spores then you have your answer for sure. It sucks to lose jars, that's why I always do a test with any syringe I use. It's hard to tell some contaminates apart from the mycelium that we are trying to grow.

Did you shake the jars after you injected the LC? That could be a reason why it's in random locations or is it just next to the glass where the liquid ran down?

Edit: are you saying that you did not use the lc but did use the same spore syringe that was used to innoculate the lc?


#5 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 04 April 2008 - 08:31 AM

Bummer... at least it was just one jar and not twenty...:dead:
If it did come from your lc then other possible contamination points are added like the spore syringe or by its injection. Trich can sporulate in an lc. I have seen it happen years ago and Lazlo was able to take a pic of it happening.
Did you shake the jars after you injected the LC? That could be a reason why it's in random locations.

Edit: are you saying that you did not use the lc but did use the same spore syringe that was used to innoculate the lc?

The rye jar was innoculated directly from the multi-spore syringe and not from the LC. So far the LC's appear fine and no trich evident anywhere - this is the same syringe I used to knock up the rye. I applied 2ml to each jar of LC and then the remaining 2ml was used on the rye experiment.
Yes, I did shake up the jars after innoculation.
As a side note...the syringe has been in the fridge for about three months since it was purchased. I used 4ml on other innoculations and capped it to be stored in the fridge since then. I can assure you there is often trich growing on shit on my fridge...usually a slice of lemon or something left over if I go away for a few days or am overly lazy will show it. So I wouldn't disregard a possible contamination of the syringe. Although...the LC's look good so far???? I'm going to test the LC this weekend and observe over the next days to see if perhaps the trich is dormant at the moment in LC but maybe will come to life upon innloculation. Is this possile or would the trich most likely appear alongside the mycelia growing in the sweet water?

#6 Mermaidia

Mermaidia

    Sober Sister

  • Expired Member
  • 4,173 posts

Posted 04 April 2008 - 08:35 AM

Maybe try pc'ing your jars longer.

I usually go 90 mins or so and when using Lazlos wbs tek i go 105min as he instructed.

Hope this helps in some way,
Merm

#7 onediadem

onediadem

    Insidious Drivel

  • OG VIP
  • 16,392 posts

Awards Bar:

Posted 04 April 2008 - 09:04 AM

Most instances you cannot see a contam in an LC. It is always advisable to do a test jar if anything is in question. In the long run this saves a lot of time and supplies.

Good luck!


#8 Lazlo

Lazlo

    old hand

  • Honorary Former Staff
  • 7,620 posts

Posted 04 April 2008 - 09:56 AM

LC's should be inoculated with agar wedges.

#9 shoe

shoe

    met 2012^23 little dudes

  • Expired Member
  • 84 posts

Posted 04 April 2008 - 02:48 PM

whoops; Did spores -> LC.

dunno if they'll germinate. I don't see why not...

#10 wildburr

wildburr

    Wildest of Burrs

  • Honorary Former Staff
  • 4,657 posts

Awards Bar:

Posted 04 April 2008 - 05:12 PM

Definitely would have PC'ed it longer (90 - 105 min) especially since your PC only gets to 13psi. I just usually soak the grain overnight, let drain thoroughly then load jars and PC, I dont cook them. Havent had a problem yet with that method.

#11 oldiebutnewbie

oldiebutnewbie

    Mycophage

  • Expired Member
  • 112 posts

Posted 04 April 2008 - 11:31 PM

As a side note...the syringe has been in the fridge for about three months since it was purchased.
-----
Presumably, these were OK.
-----
I can assure you there is often trich growing on shit on my fridge.... So I wouldn't disregard a possible contamination of the syringe.

---
Nor would I. And the contam is maybe on you and everything else in there. And concentrated too, which is the big deal I think with contams. If there is only one in the whole room, it ain't likely to ruin the project, but if there are gazillions of them, then almost certainly they are gonna get into the goodies and screw you up.
---
After 3 months, they are all over the outside of the syringes, on your hands, head, shirtsleeves, you-name-it.
------
Clean,scrub, use Betadine, build a glovebox.
----
Good luck,
OBN

#12 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 08 April 2008 - 05:21 AM

Although the syringe was most definitely surrounded by mold spores from the fridge at one point or another (By no means am I a pig but from time time something will go bad in the fridge), the syringe was sealed inside a ziploc bag and inside one of the compartments.
I also flamed sterilized the needle before innoculation.
For the sake of this trial (I ordered new syringes to start fresh), I have to assume the problem was either from an improperly sterilized grain or the polyfill filter was not thick enough and allowed contams to enter from the lid.
Since I already have fully colonized LC's from the same syringe I will be using these to innoculate the next test jar. Hopefully my assumption is correct and there are no dormant contams mixed in the LC, if not then I'll be starting from scracth with the new spore syringe (although I'll be doing this anyway).
I've soaked the rye for 24 hours, boiled till puffy but not exploded, drained and cleaned under hot water, allowed excess moisture to evaporate to an acceptable field capacity, and PC'd for 90 minutes (my pc doesn't have much pressure so I've decided to go longer).
Now my LC is awaiting it's first test this afternoon as the Rye cools down. I'm using a new syringe to pull the LC from the jar and my airport syringe is sterilized also for this process.
Also...the jar no longer has a filter as there is already a 50% volume of air contained within the jar. On my previous grows with PF type cakes in 1/2 pints I didn't use FAE and have never had a problem so I thought I would return to this principal for this trial. Upon studying some of the various teks I see there are both those who use filters and those that do not - Hippie's Rye tek for example does not have them.
Time will tell.

#13 oldiebutnewbie

oldiebutnewbie

    Mycophage

  • Expired Member
  • 112 posts

Posted 08 April 2008 - 09:18 PM

Although the syringe was most definitely surrounded by mold spores from the fridge at one point or another (By no means am I a pig but from time time something will go bad in the fridge),
------
It doesn't make you a pig, we all simply have, carry, and spread an abundance of all manner of spores,etc. I've been told that inside one's house reside vastly greater concentrations of potential contams than one will find outdoors even in the most densely populated and polluted urban areas.
-----
the syringe was sealed inside a ziploc bag and inside one of the compartments.
--
Good. It's fine. I would just wipe off the outside of ziplock with Betadine ( or Chlorox) before I opened it. Another level of caution. Probably unnecessary but.....
----
I also flamed sterilized the needle before innoculation.
---
Very good. Always works. I'm experimenting with wiping with Betadine instead but so far, it's not as reliable. Maybe my technique isn't quite good enough or maybe the wiping (which doesn't sterilise the inside of the needle) simply isn't sufficient to do the job.
-----
For the sake of this trial (I ordered new syringes to start fresh), I have to assume the problem was either from an improperly sterilized grain or the polyfill filter was not thick enough and allowed contams to enter from the lid.
---
Your logical approach is admirable. This is how each of us must struggle to incorporate the info available to us at this site into our own unique situation to reach our common goal. You'll succeed because you are narrowing down the possible problem until you isolate it. Keep on!
I had problems flaming needles and then inserting them into polyfil. I think what happened was that the hot needle created a tube (as it were) of melted poly, which allowed the meanies into my jar. I also had problems with poly plastic gunking up my needle. When possible I then switched to micropore tape and find that it works better for me. I still use poly to stuff the neck of ovenbags. Works great and prevents PC's from bursting the bag. Just didn't work well ( for me) in jar lids. Perhaps your experience will be similar. Perhaps different.
----
Since I already have fully colonized LC's from the same syringe I will be using these to innoculate the next test jar. Hopefully my assumption is correct and there are no dormant contams mixed in the LC, if not then I'll be starting from scracth with the new spore syringe (although I'll be doing this anyway).
---
So, if I understand you correctly, essentially you'll be having a controlled experiment.( I DO hope you are writing all this down somewhere to assist in your recollection.) IF your LC's are Ok, THEN it was not the previous syringe that introduced the contam. Keep eliminating.
----
I've soaked the rye for 24 hours, boiled till puffy but not exploded, drained and cleaned under hot water, allowed excess moisture to evaporate to an acceptable field capacity, and PC'd for 90 minutes (my pc doesn't have much pressure so I've decided to go longer).
----
All that sounds exactly correct. ( Well, I use cold water to rinse but......) And if onediadem thinks your grain is OK, continue as you have been doing.
-------
Now my LC is awaiting it's first test this afternoon as the Rye cools down. I'm using a new syringe to pull the LC from the jar and my airport syringe is sterilized also for this process.
---
All sounds good.
----
Also...the jar no longer has a filter as there is already a 50% volume of air contained within the jar. On my previous grows with PF type cakes in 1/2 pints I didn't use FAE and have never had a problem so I thought I would return to this principal for this trial. Upon studying some of the various teks I see there are both those who use filters and those that do not - Hippie's Rye tek for example does not have them.
----
If Hip says it, so it is.
----
Time will tell.
-----
Yup. Then we try, try again.
-----


Best of luck,
OBN

#14 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 09 April 2008 - 04:35 AM

Many thanks for those encouraging words.
One fundamental problem with these ilicit hobbies is the lack of sharing one's experiences with others. All though there may be sympathy from friends they still remain baffled by the vocabulary, my form of thinking, and of course my goals of this research. There has been many a raised eyebrow from those whom I least expected to doubt me. At best there are the chuckles at what appears to be my return to an adolescent's individualist dream. A spurt of rebellion if you will.
But then they remember who they are talking to and look at my education, my professional achievements, dedication to family, and an adherence to the mundane. At this point confusion settles in as they realize I am serious. The smile turns to a grimmace and the head tilts back ever so slightly with a doubtful eye that peers down and forward. The brow crooks upward, the nostrils open, they swallow deeply, and utter those skeptical words..."You're serious, aren't you?"
I've since realized that my adventures are not to be played out on casual listeners at a backyard barbecue!
My point is that this forum serves as a good brainstorming and idea sharing context where most of us are similarly inclined but cannot exhibit our traits in just any place. So, here's to Mycotopia and everyone who participates in this adventure!

#15 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 09 April 2008 - 05:34 AM

I've just innoculated a 1 quart test jar of rye with 4ml of live culture as specified in previous posts. I also changed my location from the kitchen to the spare bathroom for this procedure as it is much cleaner. I'm putting together a glove box for the next procedures but for now lot's of flame and alcohol wipes.
Here are some pics...note the LC close up, it apprears as a sunset seen from a distant voyage...:bow:
I'll keep you all informed as to the progress of this test jar.

Attached Thumbnails

  • DSC00012.JPG
  • DSC00010.JPG
  • DSC00007.JPG


#16 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 10 April 2008 - 05:51 PM

After 18 hours I took the following photo - colonization is already clearly underway in several locations...what a difference it makes using a live culture in place of a spore syringe!
Hopefully the meanies willn't be back to haunt me on this run.
So far so good. Godspeed ahead my fine fungal friends!

Attached Thumbnails

  • 83375d1207867442-lc-s-cool-dsc00013.jpg


#17 Hippie3

Hippie3

    DUNG DEALER

  • Founders
  • 40,642 posts

Posted 10 April 2008 - 06:44 PM

:thumbup:

#18 Triptolemus

Triptolemus

    Revivalist

  • Expired Member
  • 66 posts

Posted 14 April 2008 - 04:38 AM

Four days into incubation of the test rye jar - all is fabulous.
No signs of contamination so far. Colonization is very fast with this LC and grain. As long as there are no contams I think I'll be staying with rye as my spawn grain. To this end I have a few quesitons...
1. Does Rye typically colonize faster than PF cakes?
2. Does Rye contaminate easier than PF cakes (I've read about presoaking and endospore germination prior to pc'ing)?
3.Does rye break up easier to mix in bulk trays?
4. Does Rye grain as spawn tend to colonize bulk substrates faster than broken chunks of PF cakes?
Here are some pics of my Rye at 4 days (if there is no sign of problems by day 7 I'm going to noc up some more jars with this LC and procedure):

Attached Thumbnails

  • DSC00003.JPG


#19 sooshane

sooshane

    Hi!

  • Expired Member
  • 142 posts

Posted 14 April 2008 - 06:34 AM

Looking good on those jars - the Lc and grain.:eusa_danc What is your lc recipe?
To answer your questions.
1. Given the same volume and incubation perameters, I think that grain jars colonize a bit faster. With a Lc, 3-8 days for grain and 6-14 days for BRF. Rough estimates and depends on jar size - colonization times increase with jar volume.
2. No, Given proper sterilization, inoculation and incubation both BRF and grain jars will outgrow a contamination before it germinates equally if one is present. (Example: a bacteria).
3. Yes it does as long as you leave enough headroom in the jars. BRF and verm jars break up also if you leave out the verm barrier and only fill the jar about half way. It takes a little muscle but can be done.
4. Not really they both perform very well but the whole grain will provide more nutrition. See Brf powdered spawn and TvCasualtys Brf slurry tek for grain.




Like Mycotopia? Become a member today!