|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
deadjesusrodeo
Stranger

Registered: 10/21/13
Posts: 26
Last seen: 9 years, 6 months
|
Contamination (pics) and advice for a rookie.
#19187487 - 11/25/13 03:12 PM (10 years, 2 months ago) |
|
|
Hello everyone,
I'm writing my first post after a few months of growing. Check out this pic from my very first PF cakes:

Don't worry, my tek has improved significantly since this was taken! Read on!
And one of the larger fruits after a dunk and roll. 

Just like everyone else, I'm getting addicted to the hobby and I'm starting to scale things up a bit.
I've had a number of contamination issues recently and it's making me pretty upset. I need help finding the culprit!
After browsing here for several weeks, I'm going to try to not make any rookie mistakes. So here it goes.
Strain: Golden Teacher Substrate: Organic Rye Berries (90 minutes in the PC) Inoculation method: Grain to Grain (4th Generation from original master jar) Bottom casing: 50/50 perlite/vermiculite mix (sterilized in PC) Top casing: Jiffy Mix seed starter (sterilized in PC)
* Inoculated the 2.25 quarts of substrate on 10/20 into a single six quart container. (G2G transfer done in still air box) * Incubated at 80F until 11/4 (I have to incubate, my house stays at 65 through the winter) * Cased on 11/4 and put back in the incubator for 3 days. * Fruited into a greenhouse on 11/7. Greenhouse is well regulated with 99% RH on the bottom shelf and 85% RH on the top shelf. 72 degrees F during the day, around 68-70 at night. Light 12 on 12 off (daylight during the day, artificial during the evening). * Sterile procedure was used throughout the process using chemicals such as H2O2, lysol, 70% rubbing alcohol, and Bacteriostat (for my impeller humidifier). * I clean the greenhouse every Sunday with rubbing alcohol and I clean the humidifiers (cool mist and ultrasonic) with H2O2. * I misted twice daily with a pump sprayer until primordia/pinning was noticed on 11/19. * Cobweb mold was found on a sister container and eradicated with H2O2. * On 11/24, potentially severe contamination was noticed and treated. * Today, 11/25, about 6 very fat, dense fruits were harvested and the rest was buried outside. Upon dissection, well, you can see the rest.
TOP VIEW

SIDE VIEW

CROSS SECTION

So as you can see, pinning was less than ideal. Growth halted on the 4th day and contamination began its hostile takeover. I've read that when doing grain to grain transfers, the strain can only be stretched so far before it becomes weaker and weaker. Some recommend going from 1 jar to 10, then 10 to 100. Essentially, this batch was one step further and would fall in the 100 to 1000 range. As we speak, I am successfully growing with the 5th and 6th generation of G2G transfers.
These grains were NOT soaked and boiled before the pressure cooker, a practice that I began using on the batch following the one pictured.
The three sister containers show no signs of contamination despite coming from the same master jar. However, they all show sporadic pinning and stunted growth. Flushes from earlier generations yielded 20-30 grams, dry. These will only yield around 5-10 grams.
Trich was found on in a different container from an earlier batch but was isolated and buried before it turned green and sporulated.
I've been having major issues with contamination after the first flush and I can't seem to figure out why. Not a single tray (either in my greenhouse or in a SGFC has survived the second flush. I've tried leaving them alone, recasing, as well as dunking with the same results. My PF cakes managed to give me 4+ flushes using the dunk and roll tek.
Right now (not pictured), my recipe is 4 parts WBS, 4 parts Rye, 2 parts vermiculite, some gypsum, and 1 part BRF. Special care is taken to get the moisture level just right including rinsing, simmering, drying, then pressure cooking. I've followed multiple teks and now have a really good system that seems to work great for me. My in-jar contamination rate has been about 10%, all from wet spot bacteria. Recently, this has not been an issue, particularly since soaking and simmering grains before the PC step. I'm not sure where I read it, but someone had vermiculite in their substrate recipe so I've been doing that for a while to see how it goes.
I always use one quart of bottom casing, two quarts of substrate, then one quart of top casing. I'm fully aware of the pros and cons of casing, I prefer to use it.
Finally, I'm also having some issues getting spores to germinate on my grain. Using Z-Strain, for example, I haven't seen any signs of mycelial growth after more than a week since inoculation with 1cc of spore solution purchased from a vendor on this website.
I'll be moving to bulk substrates in the next couple weeks, but if anyone has any information, comments, or questions for me then please feel free to respond. I'm open to critique, so don't be shy! Please help me figure out why my fully colonized, cased substrates keep getting contaminated. And help me identify what's growing in the pictures above.
Thank you!
|
bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Contamination (pics) and advice for a rookie. [Re: deadjesusrodeo]
#19187690 - 11/25/13 04:04 PM (10 years, 2 months ago) |
|
|
I have some questions.
What did you spawn to, was it pasteurized, and what's your pasteurization procedure?
Did you do your g2g transfers in a SAB or LFH? How's your sterile procedure.
Why do you do this?
Quote:
Right now (not pictured), my recipe is 4 parts WBS, 4 parts Rye, 2 parts vermiculite, some gypsum, and 1 part BRF. Special care is taken to get the moisture level just right including rinsing, simmering, drying, then pressure cooking. I've followed multiple teks and now have a really good system that seems to work great for me. My in-jar contamination rate has been about 10%, all from wet spot bacteria. Recently, this has not been an issue, particularly since soaking and simmering grains before the PC step. I'm not sure where I read it, but someone had vermiculite in their substrate recipe so I've been doing that for a while to see how it goes.
That's a horrible spawn recipe(for disaster).
Take a look at these The best damn WBS prep tek. (I use rye but since you've been doing WBS just do this) Substrate tek how to pasteurize that substrate How to g2g in a SAB spawn your spawn to bulk
|
deadjesusrodeo
Stranger

Registered: 10/21/13
Posts: 26
Last seen: 9 years, 6 months
|
Re: Contamination (pics) and advice for a rookie. [Re: bodhisatta]
#19188263 - 11/25/13 06:27 PM (10 years, 2 months ago) |
|
|
Okay, this post was in regards to using rye berries as substrate (cased) without spawning to bulk. You can see from the pictures, it's just straight rye cased on top and bottom. I know the nutes aren't there for great flushes, I'm moving to bulk on my next batch.
The recipe that you quoted is for spawning to bulk, which I haven't done yet. The jars are colonizing right now (except for the spore issue I'm having) and they'll be spawned once their done. I just wanted feedback on my recipe, which is apparently a recipe for "disaster". Would you mind elaborating? I'm eager to really learn the biology behind everything, so just saying it's bad isn't going to help me understand . They seem to be colonizing just fine, I think they'll spawn well, even if it's not the best recipe.
My G2G transfers are done in a SAB. It's just a tote box. I followed this tek to the tee http://www.shroomery.org/12670/Building-A-Glove-Box-With-TheNewGuy
Sterile procedure is as good as I know how to get it. I've worked for 5 years as a paramedic, so I know a little something about sterile technique and I have all of the supplies handy. My jars have a hole drilled in the top and are stuffed with polyfil.
|
silverstem
Caps & Stems



Registered: 10/12/13
Posts: 900
Loc: jordan
Last seen: 6 years, 5 months
|
Re: Contamination (pics) and advice for a rookie. [Re: deadjesusrodeo]
#19188338 - 11/25/13 06:40 PM (10 years, 2 months ago) |
|
|
what did you case them with? mold? lol jk but really what did you use for a casing layer because growing on str8 rye berry's shouldn't look like that.
--------------------
Shroomery needs a gun forum!!!!!!!!! CAN WE HAVE ONE?????
|
deadjesusrodeo
Stranger

Registered: 10/21/13
Posts: 26
Last seen: 9 years, 6 months
|
Re: Contamination (pics) and advice for a rookie. [Re: silverstem]
#19188610 - 11/25/13 07:23 PM (10 years, 2 months ago) |
|
|
It's all written in the OP.
Bottom casing: 50/50 perlite/vermiculite mix (sterilized in PC) Top casing: Jiffy Mix seed starter (sterilized in PC)
It's just a pH balanced Vermiculite/Peat moss mix.
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 3 years, 1 month
|
Re: Contamination (pics) and advice for a rookie. [Re: deadjesusrodeo]
#19188732 - 11/25/13 07:44 PM (10 years, 2 months ago) |
|
|
My money is on either your spawn having a contamination quietly riding along on every G2G or your sterilized casing layers getting contaminated.
Were I you, I would use a bulk substrate made out of coir and/or straw and/or herbivore poo *AND* vermiculite, and if I chose to use a casing material I would pasteurize it rather than sterilizing it, which may invite contaminations due to a loss of bacterial colonization to fight off mold. I would use only one casing layer on the top of the substrate, if I used one at all.
You will find that a bulk substrate will do these things for you:
1. Improve yields per quart of spawn by stretching your grain in the same way you can use grain to stretch hamburger meat; 2. Improve yields per quart of spawn by holding more water than colonized straight grain ever could; 3. Confer contamination resistance by means not yet fully understood.
|
bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Contamination (pics) and advice for a rookie. [Re: deadjesusrodeo]
#19188942 - 11/25/13 08:35 PM (10 years, 2 months ago) |
|
|
Quote:
deadjesusrodeo said: Okay, this post was in regards to using rye berries as substrate (cased) without spawning to bulk. You can see from the pictures, it's just straight rye cased on top and bottom. I know the nutes aren't there for great flushes, I'm moving to bulk on my next batch.
The recipe that you quoted is for spawning to bulk, which I haven't done yet. The jars are colonizing right now (except for the spore issue I'm having) and they'll be spawned once their done. I just wanted feedback on my recipe, which is apparently a recipe for "disaster". Would you mind elaborating? I'm eager to really learn the biology behind everything, so just saying it's bad isn't going to help me understand . They seem to be colonizing just fine, I think they'll spawn well, even if it's not the best recipe.
My G2G transfers are done in a SAB. It's just a tote box. I followed this tek to the tee http://www.shroomery.org/12670/Building-A-Glove-Box-With-TheNewGuy
Sterile procedure is as good as I know how to get it. I've worked for 5 years as a paramedic, so I know a little something about sterile technique and I have all of the supplies handy. My jars have a hole drilled in the top and are stuffed with polyfil.
what I'm trying to say is here's how to do things right. You're not saving what you have so you might as well sit down and read the links I gave you.
I can't do your learning for you, have a read and do things in a way that works.
|
Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,855
Last seen: 14 hours, 5 minutes
|
Re: Contamination (pics) and advice for a rookie. [Re: bodhisatta]
#19189261 - 11/25/13 09:52 PM (10 years, 2 months ago) |
|
|
You're not supposed to sterilize casing material it should be pasteurized.
There's no reason for a casing layer on the bottom.
In the future if you're going to take pictures of contamination you should do it before you spray peroxide, preferably when it first appears. That would be most helpful. It's hard to tell much on a substrate that's also massively bruised and torn apart.
--------------------
|
bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Contamination (pics) and advice for a rookie. [Re: Kizzle]
#19190634 - 11/26/13 06:59 AM (10 years, 2 months ago) |
|
|
Quote:
Kizzle said: You're not supposed to sterilize casing material it should be pasteurized.
There's no reason for a casing layer on the bottom.
In the future if you're going to take pictures of contamination you should do it before you spray peroxide, preferably when it first appears. That would be most helpful. It's hard to tell much on a substrate that's also massively bruised and torn apart.
|
deadjesusrodeo
Stranger

Registered: 10/21/13
Posts: 26
Last seen: 9 years, 6 months
|
Re: Contamination (pics) and advice for a rookie. [Re: bodhisatta]
#19191535 - 11/26/13 11:33 AM (10 years, 2 months ago) |
|
|
Bodhisatta: Thanks for the links, I've read through them all and feel smarter.
Van der griegen: Thanks for the straightforward response, just what I was looking for!
Kizzle: My understanding is that a bottom casing layer acts as a reservoir for mycelial waste. With such an abundance of information on here, there are bound to be some conflicting teks. The pictures of the substrate are straight out of the container, no bruising what-so-ever. At least not from rough handling. And it was cut in half with a kitchen knife, not torn apart at all. Whatever it is is straight contam. Oh well.
So even a 100% colonized substrate is susceptible to contamination? And a non-nutritive casing layer is too?
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
|
Re: Contamination (pics) and advice for a rookie. [Re: deadjesusrodeo]
#19191923 - 11/26/13 12:58 PM (10 years, 2 months ago) |
|
|
Quote:
deadjesusrodeo said: Bodhisatta: Thanks for the links, I've read through them all and feel smarter.
Van der griegen: Thanks for the straightforward response, just what I was looking for!
Kizzle: My understanding is that a bottom casing layer acts as a reservoir for mycelial waste. With such an abundance of information on here, there are bound to be some conflicting teks. The pictures of the substrate are straight out of the container, no bruising what-so-ever. At least not from rough handling. And it was cut in half with a kitchen knife, not torn apart at all. Whatever it is is straight contam. Oh well.
So even a 100% colonized substrate is susceptible to contamination? And a non-nutritive casing layer is too?
no need for the bottom casing layer and there should be no "waiste"
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,855
Last seen: 14 hours, 5 minutes
|
Re: Contamination (pics) and advice for a rookie. [Re: cronicr]
#19192464 - 11/26/13 02:57 PM (10 years, 2 months ago) |
|
|
Quote:
So even a 100% colonized substrate is susceptible to contamination? And a non-nutritive casing layer is too?
A casing layer, even a properly made one, is more susceptible to certain molds when the spores or fragments of a nearby mold are present in large amounts. A fully colonized substrate isn't 100% immune either basically because on a microscopic level the substrate will never be absolutely 100% colonized with mycelium but contamination is rare on colonized substrates. Usually when it happens it's before full colonization is reached even though the mold may be visible until later.
--------------------
|
Psilicon
Really Nice Guy


Registered: 08/26/12
Posts: 7,057
Last seen: 3 years, 1 month
|
Re: Contamination (pics) and advice for a rookie. [Re: Kizzle]
#19193573 - 11/26/13 06:45 PM (10 years, 2 months ago) |
|
|
Quote:
Kizzle said:contamination is rare on colonized substrates
I leave all my subs flushing until they contaminate, which always happens eventually. If you're a two-flush-and-chuck kinda guy, I could definitely agree.
|
Kizzle
Misanthrope


Registered: 08/30/11
Posts: 9,855
Last seen: 14 hours, 5 minutes
|
Re: Contamination (pics) and advice for a rookie. [Re: Psilicon]
#19195232 - 11/27/13 06:38 AM (10 years, 2 months ago) |
|
|
That's a good point. I don't see much reason to keep substrates beyond the 3rd flush as yields after that are pretty inisignificant. At least if you're one who allows for consolidation before fruiting the nutrients will be pretty much used up by then.
--------------------
|
j3544956
UV



Registered: 10/23/13
Posts: 11
Loc: By the lake
Last seen: 1 year, 21 days
|
Re: Contamination (pics) and advice for a rookie. [Re: deadjesusrodeo]
#19407278 - 01/12/14 08:03 AM (10 years, 19 days ago) |
|
|
Hi
I think it might be your sub straight recipe, over kill with wbs, rye, and brf. I am currently using rye alone and that seems to work, casing I use organic vermiculite. mixing all of your ingredients can spawn dormant microbes in the grains. The rubbing alchole part, i recommend 90percent and germicidal bleach mixed with water in a spray bottle is enough. Also please do remember to wipe in one direction. It also help to pc jars, I recommend letting warm bleached water sit in your jars for an hr, than throw them into the pc.
--------------------
|
bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
|
Re: Contamination (pics) and advice for a rookie. [Re: j3544956]
#19407729 - 01/12/14 10:31 AM (10 years, 19 days ago) |
|
|
Quote:
j3544956 said: Hi
I think it might be your sub straight recipe, over kill with wbs, rye, and brf. I am currently using rye alone and that seems to work, casing I use organic vermiculite. mixing all of your ingredients can spawn dormant microbes in the grains. The rubbing alchole part, i recommend 90percent and germicidal bleach mixed with water in a spray bottle is enough. Also please do remember to wipe in one direction. It also help to pc jars, I recommend letting warm bleached water sit in your jars for an hr, than throw them into the pc.
organic vermiculite? It's a mineral that's a leftover byproduct
Mixing grains doesn't spawn microbes, time, temperature, and moisture do.
Rubbing alcohol works better at 71% than 90%. 71% is much better at killing bacteria.
You don't need to use alcohol or bleach before cooking your jars otherwise the sterilization process would be pretty useless. The PC does the work.
|
|