|Rye Grain Tips||-|
|By Ryan Waters (Zerogravity) on Thursday, January 17, 2002 - 08:14 am:|
We have a FOAF that is looking for someone experienced in rye grain spawn and how to prevent trich (green mold). They have already looked in the archives for answers. Apologies if a question is asked for which an answer has already been posted.
Please excuse the following "book" but if you know all the relevant details then it will be easier for you to isolate their problem(s).
Please note that two previous Karo Tek's were conducted using the same technique as below and no Trich developed.
They knocked up 90 - 1 1/2 litre organic rye grain jars after a successful Karo Tek with B+ & Matias. By successful they mean no contams after 14 days. Within 2-3 days around 80% of the jars contained Trich. In fact there was more Trich than Mycelium.
Here is what they did:
- Soaked grain for 64 hrs; stirred occasionally with unsterilized hands.
- Rinsed for 1/2 hour. Let stand for a couple of hours then rinsed again in small quantities for 1/2 hour.
- Let stand for 15 min to ensure all water drained
- Put grain in unsterilized 1 1/2 litre jars 2/3 rd's full. This is 1000ML of grain. Used bare unsterilized hands to fill jars. Wrapped tightly with 2 layers of foil. Lids have fiber filled holes stuffed very tight.
- PC'd @ 15lbs for 30 mins. Prior to putting on weighted scale vented for 5 mins.
- Removed jars. Absolutely no clumping. Looked dry but as this is their first time doing this they don't know what it should look like. Placed jars on counter overnight to cool down to room temp. This counter is up against a bed which is assumed to be a haven for Trich. Unfortunately jars were removed with unsterilized oven mits that have never even been cleaned.
- Created a clean room in kitchen. This involved 20% bleach solution on floors, walls, cabinets, switches & ceiling. Thoroughly cleaned range & range hood. Removed everything not bolted down. Enclosed the small kitchen in Poly and let HEPA filter run for 1 hour before they entered. Unplugged fridge as it creates a draft when it runs.
- Physical preparation included full-body disposable suits with hoods. Face mask & rubber gloves. Sprayed themselves down with lysol especially at their feet & head.
- Once in the clean room they turned off HEPA filter to prevent drafts. They moved slowly and were in the room for almost 3 hours. Sprayed the air with water frequently. Unfortunately the lids were sometimes opened too soon and this watered air got in. Once an hour also sprayed the air with a 20% bleach solution. All jars were wipped down with rubbing alcohol and sprayed liberally with lysol. The person actually knocking up the jars did only one task - touched only the needle & the lid on either the innoculation jar or grain jar - nothing else. The other person did everything else. Instead of inserting the needle thru the fiber-filled hole they cracked lid wide open. The needles were sterilized (boiled for 45 mins) and if it touched anything it was re-sterilized with rubbing alcohol. 15CC of innoculant was squirted in across the whole top layer as well as down the sides. The innoculant had been in the fridge in a vacuum sealed bag that was also wrapped in 2 bags. After injecting, the jars were shaked vigorously. Two new layers of foil were applied. Hands were cleaned frequently with rubbing alcohol.
- Jars were placed immediately in incubation chamber and not touched for 3 days. This cabinet has a HEPA filter.
-Once the contam was discovered all grain was discarded, jars sterilized, and the room incubation chamber sterilized.
They were confident they were doing everything right but to have this big a problem in such a short period of time indicates something is grossly wrong.
So what did they do wrong?
Also, they are looking for answers to each of the following questions:
1. Soaking grain. How much time is too long? (In other words can it contam sitting too long?)
2. Soaking grain. Does everything from the soaking bins to the PC need to be sterilized?
3. Rinsing. Can you do too much?
4. Draining. The grain after PC'ing and letting stand overnight looked real dry. They feel too much water is either PC'ing or evaporating out.
Do you want ALL the water out or is some in the jar good? How wet should it look?
5. Is boiling the grain after soaking redundant?
6. PC time. Is 30 min long enough for the large 1 1/2 litre jars? Can it be cooked too long?
7. When knocking up should the HEPA filter be on or should it be off?
8. Bleach residue. If you clean something that was contamed with Trich but don't clean off the bleach residue is the trich still present?
9. Dry grain. Can the grain jars be re-hydrated after being in the incubation chamber for a while if they look too dry?
10. Is Trich killed by PC'ing?
Once again, thanks in advance for your help. They could not have gotten this far without all the help that has already been extended. They hope to return the favor someday when they too are pro's.
|By relic (Relic) on Thursday, January 17, 2002 - 09:21 am:|
your problem is very simple, 30 minutes is not nearly long enough to sterilize rye. alot of people say 60 min @ 15 psi,
i always do mine for at least 75-90 min. and of course you start timing after the proper pressure is attained.
touching the grain and all that stuff prior to sterilization is absolutely fine.
i preboil/simmer(bring to boil, then lower heat to lowest setting) the grain for no more than 45 min, stirring often and keeping the water level in the pot above the grain by 1/4" or so, adding as needed. then strain the excess water off real good in a colander and then load the jars 2/3 full. p.c. for 75-90 min. @ 15 psi, then let sit in the p.c. overnight. shake the jars before inoculating in the morning or when it has cooled.
that's a real bummer, 90 jars
i feel for your friend. btw, i wouldn't apply new foil and 1 layer is really enough.
now to answer your questions
1. i don't think you can soak it too long, within reason.
4. it shouldn't look too wet after p.c.'ing. if you soaked for 64 hrs i'd say they were fine(keep in mind, i'm not a soaker). the grain can sometimes look too dry when it really isn't.
5. probably yes
6. definitely no and yes you could cook it too long though I've known of people who have regularly p.c.'d rye for 2 hrs w/ success. the big factor here is not to let the p.c. go dry. if it does, that's when things go bad.
7. off, unless it's a flowhood you're talking about.
8. no(if it is, it's dead), though bleach residue can not be a good thing to have in your jars wash them babies good w/ dishsoap and rinse them good with hot water before using them.
9. maybe, i dunno. i've never had a grain jar go dry, but then i don't have an incubator either.
|By Ryan Waters (Zerogravity) on Thursday, January 17, 2002 - 07:43 pm:|
Relic your quick, thorough, & accurate response means a lot. Thanks!
1. So you feel that, even though the large 1.5 litre jars are being used, 90 min in the PC will suffice?
2. Did you recommend not replacing the foil because it is a bad thing to do or merely because it is redundant? The reason new foil is added after knocking up is that it tears when removing it (it was really snug fitting).
3. They are still unclear on how wet the rye grain should look in the jar after PC'ing and while in the incubation chamber. When the jars were opened to innoculate, some of the grains looked very dry. In fact, there was little to no wetness on the jar's walls. It is understood that on the inside of the grain it is fully hydrated. The fiber-filled holes are stuffed tightly and are 5/16 in diameter. Once again they have never seen what it should look like so maybe they are making a mountain out of a molehill. If anyone has any pics that would be extremely helpful. (Too bad they don't have a digital camera so they could post it.)
- What happens if the grain is too wet after PC'ing?
- How fully do you drain?
- What should the glass walls look like after PC'ing?
- What about perlite in the incubation chamber?
- Do you, or anyone else, have any additional thoughts on re-hydrating the grain while it is incubating if required?
|By relic (Relic) on Friday, January 18, 2002 - 04:54 am:|
yeah, 90 min. will do
after you get them out of the p.c., the foil is basically left on as a second line of defense. to protect against falling particles onto the polyfil. i just think it's redundant to put on two layers then replace them with two new layers after inoc.
i will post some pics below to try and show the grain after p.c.'ing. notice in the pics that there is no condensation in the jars.
too wet will cause poor or incomplete colonization and can also contribute to bacterial contamination.
i put the grain into a screen type strainer and shake it up and down until no water comes out the bottom. i load the jars while the grain is still warm/hot, and when you do this you will see immediate condensation in the jars b/c of the warmth of the grain. this usually goes away. after the jars are p.c.'d and cool down. you may see condensation again when the jars are fully colonized much as you do with pftek brf jars.
depends on the temps in the chamber i guess, prolly can't hurt to have perlite in there. i just put my jars back in the box and put them away at room temps. this, of course, slows things down by a few days.
as far as rehydrating goes, I'd try it on one jar and see how it goes. though it really shouldn't be needed. use sterile water of course.
hope I helped.
|By relic (Relic) on Friday, January 18, 2002 - 05:13 am:|
few popped grains, oh well
notice, no condensation
|By Ryan Waters (Zerogravity) on Friday, January 18, 2002 - 05:15 am:|
Ya you've helped big time.
|By relic (Relic) on Friday, January 18, 2002 - 05:17 am:|
same jar as above, other side. cambo cubensis, karo injected 3 days ago.
this jar is prolly ready to be shaken, after that it will completely colonize within a few more days. i usually then shake it up one more time and wait a couple more days.
|By Ryan Waters (Zerogravity) on Sunday, January 20, 2002 - 10:17 pm:|
Wow that looks real good...I can't wait to have the same success!
|By Ryan Waters (Zerogravity) on Sunday, January 27, 2002 - 02:32 am:|
Although I find it hard to believe, my FOAF has again grown Trich. After just 2 days in the incubator green mold is visible in many jars. Too bad Trich doesn't have any market value cause he would be rich by now!
Relic they followed your advice by PC'ing the rye grain in the 1.5 litre jars for 90 mins. (There goes 4 days!) Further, in order to isolate the cause of the original failure, they prepared the second batch exactly the same way as the first batch.
They list the possibly suspicious factors common in both batches as the grain, PC duration, and inoculant. After re-reading the start of this thread, I believe there is nothing they can do to improve their sterile technique. It is for this reason that they do not suspect this. By all means, however, if anyone sees anything that they could do better in terms of sterile technique then they are all ears. Also rulled out is the incubation cabinet because the Trich is uniformly spread out in the jar from top to bottom and the cabinet was sanitized thoroughly just before the jars were placed in it.
They are forced to ask this community questions they feel are ridiculous but they are truly at a dead end street and can go no further without all you guys help.
1. Is Trich visible in liquid inoculant? This is their biggest question. They used the same inoc on both batches. They have studied the liquid as best they can and can see no contams. The myc looks perfect...healthy and lots of it. This inoc is between 3-4 weeks old. That being said, as soon as the first batch of inoc had stopped growing,it was used to knock-up the grain as well as to generate more additional inoculant. They feel that the inoc is the reason for their failure because about 3-4 weeks ago they PC'd some of the same grain for only 30 minutes (albeit it was in only 1/2 pint jars) and didn't inoculate them. Although those two jars have been sitting on the counter all this time there is no contam to report. There is no fiber filled hole, one layer of foil and even that is ripped!
2. Is Trich visible in spore syringes? Having followed everything they have read, they are wondering if the problem started with the original spore syringes.
3. Is it possible that they have "nuclear grade" Trich on the grain that isn't being killed in the 1.5 litre jars after 90 mins in the PC? In other words, are some strains of Trich not destroyed by PC'ing?
4. Are there any other contams out there that are green that this could be?
5. Could there possibly be something wrong with the grain? (Hard to believe this could be the cause when it is considered that those small 1/2 pint jars used the same stuff & have been sitting there without contams for a month now.)
6. Although it has been said that 90 mins is long enough...is it? They ask cause they were told that pc'ing destroys Trich but only when you take into consideration the size of the jar. In other words it is assumed you could PC for less time if the jar was say only a half pint and not 1.5 litres. Please Relic they mean no disrespect by re-asking a question that you were good enough to answer already
Here is what they plan to do.
- Determine if it is the inoc. They are going to use it to knock-up some of the sterilized grain that has been sitting there without contams for 3-4 weeks. If they see Trich within 3 days then that is probably the problem!
- PC some more grain for longer. They are going to PC 4 jars for 2 hours, then 4 jars for 3 hours, then 4 jars for 4 hours. Then they are going to shoot up 6 of them with the inoc. If they still get Trich then it can be assumed it has nothing to do with PC'ing or the grain.
The results of these two tests will be known by this coming Thursday. In the meantime, Quote, Nan, Relic, Ion, and other pros please read the first post on this thread and see if you can come up with what the cause of this problem could be.
Oh and just in case anyone wants to buy a case load of Trich just let me know...real low price!
|By relic (Relic) on Sunday, January 27, 2002 - 05:05 am:|
as you said, there are 3 possible problems. i'd get a known "good" syringe of spores, and try to inoc another grain jar prepared in the same manner. if it works, you'll know it was the other inoculant. if it fails you will know it was the grain(assuming it has the proper water content). are you flame sterilizing the needle before incoulating? i get mine to glow then squirt a shot through to cool, then inoc.
|By Ryan Waters (Zerogravity) on Sunday, January 27, 2002 - 06:41 am:|
Hey Relic...good to hear from ya again.
No they do not flame sterilize the needle prior to shooting each jar. The needle never comes into contact with anything but air (which is in an enclosed clean room that has been HEPA filtered. BTW the filter in the unit is brand new as well.) If it does come into contact with anything then it is cleaned with rubbing alcohol.
What are your thoughts...does it need to be flamed before each jar? This will be important to them considering the number of jars.
Thanks again Relic
|By quote: (Quote) on Sunday, January 27, 2002 - 03:23 pm:|
not really, flaming's only needed if the needle touches something dirty.
i'd sat the odds favor it being the inoculant causing problems.
you cannot see trich in syringes.how did you make your syringes ?
you should always test homemade syringes, i use a jar of pf substrate, and test each syringe by firing into 1 hole. if contams grow, i pitch it.
|By Ryan Waters (Zerogravity) on Sunday, January 27, 2002 - 08:44 pm:|
Quote...thanks again for your response.
Your thoughts are mine...they suspect the inoc.
They purchased the syringes from a reputable supplier.
(They would rather not mention them by name because they recognize that whenever there are growing problems between a newbie and a veteran, it is generally the fault of the newbie. The last thing a newbie needs is to point an inexperienced finger only to burn a bridge. If it is discovered to be the fault of the syringe supplier they would only take it up with that individual. They would however, for the benefit of this thread, mention that that was the problem but, unless you feel it wise for the Nook, not mention the supplier by name.)
1. Is Trich visible in inoc? If so what does it look like? Besides visual clues are there any other ways to detect it? They recognize that any tests would probably render it useless for knocking-up the grain but would provide answers for future reference.
2. Is there such a thing as trich that can not be destroyed by 90 min of PC'ing?
3. Are there other contams that are green besides Trich?
4. Could there be something wrong with the grain?
Please excuse their troubling you with the same questions twice.
|By Ryan Waters (Zerogravity) on Monday, January 28, 2002 - 06:34 pm:|
1. Is Trich visible in inoc? If so what does it look like?
2. Is there such a thing as trich that can not be destroyed by 90 min of PC'ing?
3. Are there other contams that are green besides Trich?
4. Could there be something wrong with the grain?
|By quote: (Quote) on Tuesday, January 29, 2002 - 02:35 pm:|
2. possible, some endospores are very tough.
4. yes, and that's the most likely culprit, assuming your syringes were good.
i'd find a new source.
|By Ryan Waters (Zerogravity) on Tuesday, January 29, 2002 - 06:59 pm:|
Thanks Quote...I'll let ya know when they have answers to conclude this thread.
|By Eatyualive (Eatyualive) on Wednesday, January 30, 2002 - 02:16 am:|
Damn this is a detailed post. Many, many questions. but thanks for the good response guys! i always have a bitch of a time with rye. it seems that even when my jars fully colonize ( sometimes less than a week) i always have the mycelia stall out and stop growing near the end after the last shake. i have had success with koh samui. i actually had 5 flushes and then it contamed but it was hell gettin there.
|By Ryan Waters (Zerogravity) on Monday, February 11, 2002 - 11:12 pm:|
Wow! Here's a puzzle for all you veterans. Of course with every good game there is a catch...you gotta read the "novel" I wrote.
My FOAF got trich for the third time! Could hardly believe their eyes! Hard to imagine this is possible when I consider their procedures and how much time, money, & effort has gone into it. They are definetly confused but no where near ready to give up.
Here's the update:
- They knocked up rye grain that had been sitting for over a month without contams. It had only been PC'd for 30 mins but mind you they are only 1/2 pints. Saw trich at 2 days so they knew it was the inoc.
- Ordered fresh spores from the SporeLab. BTW extemely quick response & delivery...3 days! He emailed me to say when he received the $ and when exactly he was mailing...cool!
- PC'd more grain but this time only for test purposes as per Quote's good advice. 4 jars for 2 hours, 4 jars for 3 hours, & 4 jars for 4 hours. No visible difference between jars. Let these jars sit at room temp for 10 days. No contam developed.
- Made up more inoc using the method described in agonizing detail above . Wow...the jar was FULL of myc within 2 days!
- Knocked up 3 of the jars that had PC'd for 2 hours. Trich detected at 2 1/2 days. Of course just like the other times there was a good deal of myc.
Because the jars were sterile (soaked in bleach then cleaned with soap) & the syringes new, the process of elimination says it must either be
But how could that be? If Max was having a problem I'm sure he would not have sent spores out. Even if he didn't know at the beginning, he would have found out by the time they had ordered 3 times from him cause other people would have sounded the whistle.
It had better be something as obvious as the nose on their face cause it looks to me they're pretty good at this.
They conducted 2 tests with the sterilized grain sitting for 1-4 weeks without contam. Also they are getting it almost fresh from Nunweilers which are a huge & reputable mill/brand.
My FOAF is embarrassed to report these attempts to this community. They feel they will become unwelcome here cause they only take and never give. They want the pros to know they take this very seriously and are very grateful for all the excellent advice all the busy pros have given. Any advice they have been given in the past they have followed. They have followed all the advice they have read here to the letter but still are missing something huge. They have learned 95% of what they know here. They are desperate for any good advice.
Could you please take a quick moment and tell them what you would do if you were them?
|By relic (Relic) on Tuesday, February 12, 2002 - 01:11 am:|
sounds like you have a green thumb ;)
sounds like the grain. you didn't say what happened with the jars cooked for 3 and 4 hours??
i am dumbfounded. rye grain has been a cinch for me since i tried presimmering for 45 min. then p.c.'ing for 90 min. @ 15 psi. i dunno what to say. you tested the syringe and it worked. so, it must be the grain.
|By Ryan Waters (Zerogravity) on Tuesday, February 12, 2002 - 02:52 am:|
Your right relic...it really seems it should be simple. And this is supposed to be the easy part...one wonders what casing will be like!
My reasoning means little, but help me to understand why you mention the grain. The jars that sat there for 4 weeks didn't contam. The jars that got PC'd for 4 hours then sat in the same location as the others for 10 days didn't contam either but the moment they got knocked up they contamed. The jars they shot up had been PC'd for 2 hours. How could anything on the grain survive that?
The jars that got PC'd for 3 & 4 hours are still untouched & uncontamed. Tonight they're going to shoot a test 3 & 4 hour jar with 35 cc's of RO water. That will answer the question if the jars are not contaming cause they are too dry. Note however that they do not look too dry. A long shot but at this point they are considering everything including the position of the planets.
Do I sound as if they're making light of your idea? I am not. They will try another mill but there are only two mills in the country that have this organic rye. They are not buying from a retail outlet. It is just that no one has reported having grain trouble of this nature after PC'ing properly.
Do you see anything they could do better in their sterile technique? When working in the small Clean Room which was HEPA filtered for 1-2 hours, do you see a problem with spraying the air with either straight water or a 10% bleach water solution? They do this cause for the amount of time they are in there with the HEPA off they feel that airbourne contams could develop. But what if the wet air drags down something in the air and onto the syringe or jar? This would not explain a 100% contam rate 3 times. Is this question too desperate?
They have an uneasy feeling that it is the spores. After all Trich is not visible in syringes and that is one of the constant factors. Three different loads of grain all 100% contam within 60 hours of shooting. It seems the most likely. If it was unsterile technique then they had better be commiting some huge mistake for it to affect things 3 times. This particular supplier isn't PF but it is on Quote's or Nan's top 4 list. If that is the problem they are sure that the supplier doesn't know...people are probably afraid of pissing the guy off by telling him. After all telling the guy won't fix the situation. Kinda like "winning the battle but losing the war". However the case could be made that if that is the problem then others should know. Hmmmmmm. They would probably let him him know after they know for sure. This would involve trying spores from another supplier and changing nothing in their technique. Then if they get the same results they can inform him and it will then be his responsibility to inform our community.
At the same time, both B+ & Romero strains were shot...seems remote that 2 different strains in a total of 9 syringes over a period of 2-3 months would all be bad.
What are your or anyone else's thoughts? I am sure that there are a lot of veterans reading this that have comments...please bring it on!
|By relic (Relic) on Tuesday, February 12, 2002 - 04:14 am:|
i thought you said you shot a test jar that germinated good? i am getting a bit confused. are you using a vendor syringe directly, or making some karo type inoculant out of it first? btw, i only flame sterilize before the initial jar, not inbetween jars unless the needle touches something besides the polyfil.
|By Ryan Waters (Zerogravity) on Tuesday, February 12, 2002 - 05:37 am:|
- No, there have been no successful jars...100% failure every time...which is a helpful indicator cause whatever thay are doing wrong they have been doing it since the beginning.
- They are using a vendor syringe and shooting it into a Karo jar to grow it out. Then when there is significant myc they are then shooting the grain jars.
- Ya,they knew what you mean't re flame sterilization
|By relic (Relic) on Tuesday, February 12, 2002 - 08:22 am:|
The karo inoculant could very well be the problem here.
have you modified the karo tek in any way? anything? vented or sealed karo jars?
are you using a glovebox to do the karo?
if so, are you using latex or similar type gloves within it?
have you tried shooting the rye directly with spores at any point?
you said you did the karo twice before and it worked, i assume with brf pf jars?
did you change anything in the karo preparation from those times to these?
|By quote: (Quote) on Tuesday, February 12, 2002 - 03:49 pm:|
one way to test if the grain is the problem or not would be to make a few pf style jars and inoculate them, if they turn green, then it must be the inoculant instead of the rye.
then, to finish the process of elimination, test one of the suspect syringes directly on a jar, no karo tek, if it comes up clean, it's not the syringe but your karo tek to blame.
|By Ryan Waters (Zerogravity) on Tuesday, February 12, 2002 - 08:47 pm:|
Thanks Relic. I see where you are going.
They have done the Karo Tek 3 times and each time the result was the same. They got lots of myc within the expected time frame. No contams were ever visible but Trich isn't visible in inoc anyways. They changed the tek on the 2nd & 3rd time but not in a way that they can see would develop Trich.
But first, here is what they did the same for all 3 times.
- RO water, Karo syrup, and 3 marbles were PC'd for 45 mins @15 psi. The jars were sealed and had 2 layers of foil. It was allowed to cool down on its own while in the PC without lifting the weighted scale while under pressure or lifting the lid when there was no pressure. (Doing otherwise would smash the jars and be dangerous.) Because it is not required, the solution prior to PC'ing was not prepared in a Clean Room. However, many steps were taken towards cleanliness.
- The next day a Clean Room was made. (See original posts above for details.) Even though they are newbies, they have no real concerns re proper sterility for each of the 3 batches. Why? They are fans of the Nook! (Is that considered shameless brown nosing?)
- Injected one whole shaken reputable (but not PF)vendor syringe (B+ & Matias Romero) into a solution of light Karo Syrup & RO water in a 1.5 litre sealed jar. 2 new layers of foil were applied.
- Jars were then placed in an dark sterile incubator set at a stable 84 degrees. The cabinet is in a HEPA room. Jars were not touched, shaken, or looked at for 2-3 days.
This is how the tek varied.
- 1st time
Ingredients: Used the recommended amounts of water & syrup. This is 1000ML water & 1/4 cup light syrup (which is 4%).
Shaking: Every 2-3 days & vigourous.
Results: Records were not taken but if memory serves correctly, the first myc was seen at around 4-5 days (?). It was then shot at the 14 day mark when there was lots of myc.
- 2nd & 3rd times
Ingredients: Things were doubled...500ML water & 1/4 cup Karo. Half the water was used so that there would be more oxygen in the jar & twice the Karo for more food.
Shaking: As per Ion's suggestion, the jars were shaken immediately after inoculating (in the Clean Room) a couple of times with the lids coming on/off so that there would be more disolved oxygen in the solution. Once in the incubator they were of course left sealed but instead of shaking every 2-3 days they shook vigourously everyday.
Results: Lots of myc ready to shoot within 2-3 days.
Please note. The original Karo Tek was followed to the letter and it worked fine. They only were experimenting by changing the amounts. Until they get word from the pros, newbies should follow the recommended tek.
Does this community see any mistakes here?
|By Ryan Waters (Zerogravity) on Tuesday, February 12, 2002 - 08:54 pm:|
ooops forgot. No Relic they have not shot spores directly onto rye grain...but will. Good piece of reasoning.
|By Ryan Waters (Zerogravity) on Tuesday, February 12, 2002 - 08:55 pm:|
That's great advice Quote. A good procedure to isolate whether it is the spores, inoc, or grain by changing only one thing at a time. They will get on that immediately and post the results ASAP.
|By Ryan Waters (Zerogravity) on Tuesday, February 12, 2002 - 10:25 pm:|
Hey, what type of rye grain are you pros using?
I ask cause there 3 types of rye grain according to the Nunweiler's rep.
a.) organic, which is no additives or preservatives
b.) non-organic, which has additives but no preservatives
c.) non-organic, which has additives & preservatives.
They are taking a long shot guess that if people are successfully using type b it may be working well because the additives somehow are trich unfriendly but myc friendly.
Continues in: Pros Debate: Oven Tek vs Glovebox
|Posted by: ninja Mar 19 03, 01:21 AM GMT|
| I have a friend that has
worked with several types of cultures in their life. Recent total disaster from contamination has lead to a fresh start. She had one mush after the episode, a gulf coast, and the cap was only the size of a dime, actually a little smaller. It was placed in a 1/2 pint jar for printing and left for over 24 hours. From that single minute print, a 12 cc syringe was made, using distilled water. The syringe was placed in the refrigerator for 3 days. Previously there were 12 - 1 quart jars pf rye that she had pressure cooked. Being the first shot, she decided to keep the grain a solid measure and vary the water content due to not knowing the moisture content of the grain. 3 of each type of jars were made up. 3/4, 1, 1 1/4, and 1 1/2 cups of water to one cup of rye, vigorously shaken etc before and after cooking.
The 1 1/2 v water jars seemed too wet, and the 1 1/4 weren't far behind, they had overcooked, and some of the rye broke open and most of it stuck together in those jars.
one of each jars were innoculated with a piece of a cake that wasn't expected to make it. The mycellium started growing after 2 days, then the jars started venting peutred stinky vile and vulgar fumes... The growth halted, and has shown no signs of progress since, they are considered a failure.
48 hours ago two of each of the 3/4, 1, and 1 1/4 water jars were innoculated, using the entire 12cc syringe from the miniature print.
The tops of the jars were sterilized with a torch, and so was the drill, then a 3/8 inch hole drilled in them, in a very clean setting, Polyfill was stuffed in the holes, then they were innoculated.
Less than 24 hours after innoculation there was good signs of growth. At 48 hours, there is about 10 percent coverage in the higher water content jars. Just one thing... The stink.. It has returned. The incubator is sealed and has a gentle heating system maintaining a stable 82 - 86 degrees Fahrenheit.
Questions if anyone has been here.
1. Is the bitter sweet stink of th grain normal, pf cakes don't smell anything like this?
2. If it isn't could it be caused from cooking the jars too long?
3. Do the figures of water seem right for the one cup of rye?
4. When cooking at 15 psi, what is the least time you can cook and and kill the baddies in the grain?
5. Should one pre soak the grain ( to germinate any unwanted spores) then pressure cook it, if so, does this reduce the cooking time.
6. when casing the grain after the mycellium has grown properly, what works best, plain verm on the bottom then grain then more verm?
7. With the poly fill in the lids, should the ambient humidity be boosted in her incubator?
For her sake, I would appreciate if only those whom have worked with this in the past, or seen it first hand by someone elses working with rye.If you can help her out. Any suggestions will be forwarded to her though. Thanks for any help people, I don't work with grain and could not answer her questions nor does she have web access. I am sure if she ever does, this is gonna be one of her first stops.
|Posted by: Mycota Mar 19 03, 01:53 AM GMT|
| Not much help, as I hate rye.
The reason I hate it is, the moisture content of rye can vary by about 10%, right out of the bag, or bin. Before you try increasing it, via one method, or another
So, no set amount of water always works. I tried rye spawn jars about 6 times. Of those 6, it worked twice. A couple were to wet, another clumped hard & one was too dry.
Then, I tried white millet & had about the same luck.
Then I came up with a WBS tek. It is easy & forgiving.
Rye has a unique smell, but not like you describe.
Next time try WBS, soak overnight, rinse, drain (well), fill jars 3/4 full, use a tyvek filter, one 3/8ths or two 1/4 inch holes (predrilled) in lid & PC 60 to 90 minutes @ 15 lbs. Shake while hot, cool to room temp & you are good to go.
|Posted by: Mycota Mar 19 03, 02:01 AM GMT|
|Posted by: steveoi812 Mar 19 03, 02:03 AM GMT|
| works like a charm
easy 2! Did I forget to mention dirt cheap?
|Posted by: DirtyWOP Mar 19 03, 08:13 AM GMT|
| If it smells bad, your jars are probably contaminated with some type of bacteria.......
I think rye is fine.
I prefer to hydrate it before-hand, by soaking or simmering in hot water for an hour. Soak for 24h. It is too easy to get the water content wrong when you add water directly to the jars, not to mention all that sticky starch and flour doesn't get rinsed off....
the idea is to get the rye to the point where it is as swollen as it will get before you spoon into the jars. If you can easily mash the grain between two fingers, but the grains aren't bursting yet, it is ready to PC.
And add your filter material before you sterilize next time.
|Posted by: DirtyWOP Mar 19 03, 08:17 AM GMT|
verm is OK, but most prefer to case with a mix or 50/50 verm/peat moss, or 60/40 verm/coco coir
As long as your filters aren't too big, and your incubator isn't dreadfully dry, it should be ok. Keep an eye on the dryness. Mycelium on rye will take on a blue tinge if it's too dry. A trick I sometimes try when jars get too dry (only if there is already mycelium growing) is shooting 3 mL of peroxide into the jar. This will help ward away contams and decompose into extra water and O2 for the culture to utilize.
|Posted by: ninja Mar 19 03, 10:25 AM GMT|
|Thank you to both of you!! Your time is appreciated. Less than 72 hours later after the innoculation, I have 2 jars @ 25 percent or better, it is unreal, just madd. I have't seen anything as aggresive as this in all my life.|
|Posted by: ninja Apr 28 03, 03:50 AM GMT|
| ok peeps, I wasn't meant to make this stuff happen here, for there has beena problem. I figured it out though. About half way through the process of spawning the mycellium, the rye decided to sprout!!!! This is the deal, I know it can't sprout after i pc it, so the stuff was germinating when I was soaking it. Any how, it made a big ole stinkin mess of a bunch of jars, and I am not gonna go on with this till later. I would imagine if I pc the rye before soaking it, it can't sprout, and this shouldn't effect the process with the mycellium. Thanks again for your help people.
|Posted by: OZZ Apr 28 03, 06:47 AM GMT|
|1 cup rye to 3/4 cups water, but no more water than that ..... usually get 5 +/- quarts out of 7 to colonize completely with these figures. Soak overnight, PC for 90 minutes, shake, let cool, inoculate, incubate, spawn or case|
|Posted by: dcyans Apr 28 03, 07:07 AM GMT|
|Posted by: DirtyWOP Apr 28 03, 07:42 AM GMT|
it's just like any grain if you soak it
you can take almost any grain that is large enough
and get the moisture content perfect....
as in 100% success
if you would just soak it before-hand, like birdseed
WBS isn't the only grain that you can soak
You just have to get a feel for how a grain feels between the fingers when it is ready....some take longer than others to swell.....
Wbs takes 1 day, rye takes 2, popcorn takes 2 or 3 days and some boiling to get right
edited to add that a consistant PCing time, and TWO standoff plates is necessary for consistant results as well
|Posted by: DirtyWOP Apr 28 03, 07:48 AM GMT|
| It should not be sprouting.....
you may have soaked for too long
before PCing discard any that have germinated
Inoculation Volumes for Grain Jars : Shroom Glossary