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YaMoonSun
The Double Standard


Registered: 10/23/14
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Loc: NY
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Contaminated, again; Six out of Six
#22409771 - 10/20/15 03:18 PM (8 years, 3 months ago) |
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...So I made up 6 pf cakes using rye flour, and all of them smell of spoiled milk. There's life in 4/6 of the jars, but I don't have much faith in them. Where did I go so wrong?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22409797 - 10/20/15 03:26 PM (8 years, 3 months ago) |
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How did you sterilize? Rye flour will contain a substancial number of endospores compared to brf which will have very few. Because of this I would want to PC cakes with rye flour for 90 min to be safe.
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Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22409800 - 10/20/15 03:27 PM (8 years, 3 months ago) |
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Quote:
YaMoonSun said: Where did I go so wrong?
Explain your process. No way to tell you what you did wrong from your post.
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Tehbearz

Registered: 11/04/13
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Re: Contaminated, again; Six out of Six *DELETED* [Re: YaMoonSun]
#22409803 - 10/20/15 03:27 PM (8 years, 3 months ago) |
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Post deleted by TehbearzReason for deletion: I want it deleted
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Handslikehouses
Lion Skin

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Re: Contaminated, again; Six out of Six [Re: Tehbearz]
#22409807 - 10/20/15 03:28 PM (8 years, 3 months ago) |
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Don't give up man. Don't give contams the satisfaction.
-------------------- Considering spores can survive the vacuum of space, I would assume it's safe to say that mushrooms are on every planet waiting for the right conditions to present themselves. What it all boils down to, is aliens are on shrooms.

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Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
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What's the timeline on the contamination? I have had some trouble with steamed rye flour based PF substrate. If you're getting decent colonization (say 50%+-25%) then stalling, that sounds more likely to be endospores. If you aren't getting much of anything, it's probably inoculant.
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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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YaMoonSun
The Double Standard


Registered: 10/23/14
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Re: Contaminated, again; Six out of Six [Re: Pastywhyte]
#22409838 - 10/20/15 03:37 PM (8 years, 3 months ago) |
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I followed my standard procedure of steam sterilization in a large pot for 1h30m to 2h30 depending on the size of the jar. I've yet to acquire a pressure cooker. I should probably put that on my Christmas list
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YaMoonSun
The Double Standard


Registered: 10/23/14
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Re: Contaminated, again; Six out of Six [Re: Tehbearz]
#22409846 - 10/20/15 03:38 PM (8 years, 3 months ago) |
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No, I disposed of that LC, it was filthy.
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Jerkypall
Stranger

Registered: 05/21/15
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22418388 - 10/22/15 01:46 PM (8 years, 3 months ago) |
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Just use BRF with a Pressure cooker. Never have I had one contaminate, so long as you let it cook for a while.
Sincerely, Jerkypall
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YaMoonSun
The Double Standard


Registered: 10/23/14
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Re: Contaminated, again; Six out of Six [Re: Jerkypall]
#22418393 - 10/22/15 01:49 PM (8 years, 3 months ago) |
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 ..Yeah man, been meaning to do that for quite some time now
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YaMoonSun
The Double Standard


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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22418550 - 10/22/15 02:27 PM (8 years, 3 months ago) |
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So should I let the mycelium continue to try to colonize the smells bacterial jars? Or just toss them now and start over? Basically, avoid rye flour until I get a PC, yeah? Or bump up my times to like 3h30m?
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JacobStorm
psychedelic cartel



Registered: 07/21/15
Posts: 1,499
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22418565 - 10/22/15 02:29 PM (8 years, 3 months ago) |
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I personally would start over. I don't think increasing the time is going to help? Something smells fishy here and I don't think it's the stove cooking.
-------------------- Rogger Rabbits Mushroom growing videos Ethnobotanical garden forum Inocuole tea TEK azur said: If you like 6th grade results, then 6th grade projects are great.
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YaMoonSun
The Double Standard


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Re: Contaminated, again; Six out of Six [Re: JacobStorm]
#22418587 - 10/22/15 02:34 PM (8 years, 3 months ago) |
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Smells more like spoiled milk to me. Could really use some LI in my life
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Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22418675 - 10/22/15 02:54 PM (8 years, 3 months ago) |
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If you can't get a PC, can't get BRF (check healthfood stores) it might just be best to drop 10-15$ on a couple bags of sterilized rye. Assuming your inoculant is clean (if it's from a reliable vendor it probably is) you could probably save a lot of time and a lot of vendor quality inoculant.
Add a $5 tote and a brick of coir $2.50, and you can prolly pull at least 4 zips for a total of like 20-30$. Sounds fair.
Better than paying .50c an oz or whatever for stupid Bob's Red Mill Rice Flour. BRF is about 2.50$/lbs at a local healthfood store for me, but I live in a Global Center of Diversificated Multiplural Metrohipster.
 inb4 sponsor.
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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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JacobStorm
psychedelic cartel



Registered: 07/21/15
Posts: 1,499
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I got six pounds of general mill BRF for 18 bucks. But then again I work at a grocery store and got that at cost price.
-------------------- Rogger Rabbits Mushroom growing videos Ethnobotanical garden forum Inocuole tea TEK azur said: If you like 6th grade results, then 6th grade projects are great.
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YaMoonSun
The Double Standard


Registered: 10/23/14
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I generally just buy the whole brown rice and grind it finely in a coffee grinder, but I decided to swap for rye flour, and I've had a fuck-load of contaminates ever since. I was going to blame the vermiculite at first, but yeah. Until I go to the store, I'm going to fuck around with a "new" substrate that I'm curious to see colonized.
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YaMoonSun
The Double Standard


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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22449816 - 10/29/15 04:16 PM (8 years, 3 months ago) |
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Happened again using BRF, so I made a new syringe, and remade the jars for a third imte
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Zuul
Gatekeeper


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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22449914 - 10/29/15 04:37 PM (8 years, 3 months ago) |
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Are you reusing the same jars over and over? With all the different variables it's not going to be easy trying to pin point the cause of contamination. If these cakes don't make it toss everything, get a PC, and get fresh everything. There's a turd in your punch bowl and it's not going to be easy fishing it out.
-------------------- Nothing in this world can take the place of persistence. Talent will not: nothing is more common than unsuccessful men with talent. Genius will not; unrewarded genius is almost a proverb. Education will not: the world is full of educated derelicts. Persistence and determination alone are omnipotent. Calvin Coolidge
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mushpunx
Fungus Punk



Registered: 04/20/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22449915 - 10/29/15 04:37 PM (8 years, 3 months ago) |
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I have never had a cake contaminate before. And Ive never inoculated one inside an SAB only open air.
Id say most of the time cakes contaminate either from wiping the needle down with ISO *after* flame sterilization and the rest of the time its from a bad syringe.
Seeing as you are making your own syringes I would almost gaurentee there is a flaw in your printing or syringe making.
Can you please walk us thru both? Sterile technique included
Working with agar and grain you quickly realize just how often syringes aren't clean enough. Rarely are they clean enough for grains, but can be clean enough for cakes.
--------------------
 Amateur Mycologists United AMU Q&A
Edited by mushpunx (10/29/15 04:40 PM)
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YaMoonSun
The Double Standard


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Re: Contaminated, again; Six out of Six [Re: mushpunx]
#22450015 - 10/29/15 04:57 PM (8 years, 3 months ago) |
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As I was pouring them into the garbage outside, I smelled the dry layer, and it smelled awful, but the substrate below didn't smell as bad.. makes me wonder if I just tossed good jars.. I mean, am I supposed to wait for the jars to turn mushy and gross before I toss them? Maybe using your nose isn't the best idea? I didn't see any signs of mycelium, but it was only a week after inoculation with ms syringe, so 
Quote:
Are you reusing the same jars over and over?
Yes, after a proper washing and steam sterilization 
My print making is pretty basic stuff, I just collect a mushroom that I harvested and twist the cap off gently; Then I put the mushroom gills downward on a piece of foil and cover it with a clean glass for 24 hours.
The syringe making I looked up on a YouTube video; I just don't have a flow-hood, so previously I was making them in my room, and the a/c wasn't working so there was no air-flow in the room if no-body opened the door. Now I have to use a SAB because the damn vents never turn off and people keep the fucking door and windows opened all the time because they love being cold..
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YaMoonSun
The Double Standard


Registered: 10/23/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22450020 - 10/29/15 04:58 PM (8 years, 3 months ago) |
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Watch my cup of noodles be the only one that doesn't contaminate
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mushpunx
Fungus Punk



Registered: 04/20/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22450156 - 10/29/15 05:26 PM (8 years, 3 months ago) |
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Are you printing in your still air box?
You want to prep the SAB by spraying the walls lightly with soapy water. Wear freshly laundered clothes, tyvek sleeves if you have them, gloves, scrub arms like a surgeon, face mask, wipe gloves down with ISO. You need to know how to use an SAB to utilize it properly, you never move anything over top sterile work, use controlled desicisive hand motions to keep the air as still as possible.
For printing you need to tear off a piece to get to the clean foil. Cut the cap with a sterile scalpel, fork the top... place onto foil, after 24 hrs remove cap , let dry in still air box, carefull place in fresh (fresh is sterile) ziplock. Most people fold in sterile foil envelope first.
That you looked up the syringe making on YouTube is not helpful. I asked you to describe your process.
We need to know that you are properly sterilizing your syringes, water, tools etc.
You need to sterilize a shot glass, wrapped up in foil so that it can be opened in your still air box. The syringe should be filled with water and sterilized wrapped in foil as well. A jar of water should also be sterilized and allowed to cool. Some people add a couple drops of sex lube in prior to sterilization.
You carefull open your print in still air box following the tips I gave you. You would flame sterilize your syringe and drop a couple drops onto the print where you intend to use, and with a flame sterilized loop loosen up that section, and pour off the print into the shot glass with however much sterile water in it. Flame the syringe and pull up the water and cap.
--------------------
 Amateur Mycologists United AMU Q&A
Edited by mushpunx (10/29/15 05:39 PM)
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YaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
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Re: Contaminated, again; Six out of Six [Re: mushpunx]
#22450253 - 10/29/15 05:46 PM (8 years, 3 months ago) |
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No, I didn't print in the SAB, the point of the glass was to prevent air flow 

My SAB is pretty whack It probably needs to be redesigned. It's just a temporary fix to prevent contamination from getting into the sterile shot-glass. The syringe is already made
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mushpunx
Fungus Punk



Registered: 04/20/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22450278 - 10/29/15 05:51 PM (8 years, 3 months ago) |
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Quote:
YaMoonSun said: No, I didn't print in the SAB, the point of the glass was to prevent air flow 

My SAB is pretty whack It probably needs to be redesigned. It's just a temporary fix to prevent contamination from getting into the sterile shot-glass. The syringe is already made 
Covering the print is fine, but what about all the contams landing on your foil before you even put the cap down? Printing in open air is only suitable for prints that are going to be cleaned up on agar.
Well Im almost certain your contams are coming from your syringes, friend. Unless you walk us through how you are printing and syringe making step by step I doubt we can help you solve your problem
--------------------
 Amateur Mycologists United AMU Q&A
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YaMoonSun
The Double Standard


Registered: 10/23/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22450313 - 10/29/15 05:59 PM (8 years, 3 months ago) |
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Quote:
I asked you to describe your process.
Usually, with success
• Boil water for ten minutes to sterilize • Draw sterile water into the syringe and expel into nearby glass eight times • Draw sterile water into syringe once more, and store it in the fridge to cool • Use 70% isopropyl alcohol to sanitize a shotglass using half a paper towel, drying with the other half • Flame sterilize a blade, one that won't tear the aluminum • Scrape a few spores into the shotglass • Flame sterilize the syringe and eject the sterile water into the spores, and suck them back into the syringe quickly, but accurately, as not to get any bubbles.
The print making doesn't have but three steps... I already told you what they were. The SAB was made last night; Hopefully that solves all the problems
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Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
Posts: 3,038
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Re: Contaminated, again; Six out of Six [Re: Zuul]
#22450331 - 10/29/15 06:03 PM (8 years, 3 months ago) |
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Quote:
Zuul said: Are you reusing the same jars over and over? With all the different variables it's not going to be easy trying to pin point the cause of contamination. If these cakes don't make it toss everything, get a PC, and get fresh everything. There's a turd in your punch bowl and it's not going to be easy fishing it out.
...Sterilizing the substrate sterilizes the jar...
I agree with punx, it's probably your syringe-making that's the problem. PF tek was designed for open air inoculation. Are you using fine verm for the barrier? If not, you might wanna invest in some silicon sealant and make injection ports and a tyvek covered GE hole. It's only like $3 at ace, tyvek free at the post office.
Definitely sterilize whatever you scrape spores into. The guide I read (off this site's tek section) said to whipe it down with alcohol. Also, when you lower the syringe to suck up spores, tilt the container sideways so anything falling off your hands or the syringe falls on the side of the glass or the floor of the air box. I'm surprised my first grow wasn't a flop. Sounds like you read the same guide I did. Whipe your syringe down as well, or just put it in a piece of foil in your steam sterilizer.
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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22450537 - 10/29/15 06:54 PM (8 years, 3 months ago) |
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YaMoonSun said:
Quote:
I asked you to describe your process.
Usually, with success
• Boil water for ten minutes to sterilize • Draw sterile water into the syringe and expel into nearby glass eight times • Draw sterile water into syringe once more, and store it in the fridge to cool • Use 70% isopropyl alcohol to sanitize a shotglass using half a paper towel, drying with the other half • Flame sterilize a blade, one that won't tear the aluminum • Scrape a few spores into the shotglass • Flame sterilize the syringe and eject the sterile water into the spores, and suck them back into the syringe quickly, but accurately, as not to get any bubbles.
Here's one of your main problems. You have the term "sanitize" correct.. you arent sterilizing the shot glass. If you dont believe me place a drop of ISO on an agar plate and watch what grows. Even if you did sterilize it, touching it with a paper towel to dry it un sterilized it.
I believe that is where the bulk of your contamination is coming from. We usually wrap the shotglass up in foil and sterilize in a pressure cooker cycle, then only open the foil in still air box.
Another less serious issue is scraping spores with blade. It fucks up the spores. Loosen thrm with a few drops of sterile water (on print in still air box) scrape them up with a flame sterilized loop (easy to make) and pour the spores/drops of water into the sterile shot glass.
Any lapse in sterile technique is enough to doom your syringe. Moving your gloves over top your open print or over your sterile shot glass can knock contaminants in. Even disturbing the air too much can move contams to settle into exposed sterile containers.
I dont know if there is a way to steam sterilize your shot glass PF style, or if you need to invest in a PC (which you most definitely should anyways), but if you correct these couple things it should go a long way.
I would toss any syringes you have already made dude
--------------------
 Amateur Mycologists United AMU Q&A
Edited by mushpunx (10/29/15 07:05 PM)
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Supalemonhaze
Spore syringe hater.



Registered: 10/02/15
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Re: Contaminated, again; Six out of Six [Re: JacobStorm]
#22450561 - 10/29/15 07:03 PM (8 years, 3 months ago) |
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Yea id definitely switch to brf if I didn't have a pc. One of the most forgiving substrate there is. Shouldn't have much trouble finding it. If I get it in my couuntry its a safe bet you'd find it anywhere else
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MadSeasonStudent
Enjoying Life



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Re: Contaminated, again; Six out of Six [Re: Supalemonhaze]
#22450618 - 10/29/15 07:17 PM (8 years, 3 months ago) |
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You should of made up some agar or brf to test first. I had a foul smelling cake that smelled like sour milk. I left in water for a week. I wouldn't fruit it. Try leaving it longer on the stove next time.
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YaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
Loc: NY
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Re: Contaminated, again; Six out of Six [Re: mushpunx]
#22450677 - 10/29/15 07:30 PM (8 years, 3 months ago) |
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Bulk contamination came from sterilizing the shot glass wrong. Had never sterilized before and wanted to be extra causious that time around, and I goofed I guess. Had no prior issues with just sanitation. Also, yes, I swapped to brf, in regards to what haze said, and mad has a student? I'll be damned. I know dude, agar, I need it badly.
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Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22450716 - 10/29/15 07:38 PM (8 years, 3 months ago) |
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Probably want to synch that foil down like a doo rag with some string. Can you even do agar without a PC? Maybe there are some microwave methods, I'm not sure. I summon thee lord blackout to lay down your microwave wisdom on all us ignant bitches.
IME grains and cakes are about the same from spore syringe. If your inoculant's good, your grow will be good.
--------------------
I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
Edited by Machiavelliavore (10/29/15 07:39 PM)
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
Last seen: 11 days, 14 hours
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A syringe is rarely clean enough for grains man. Its why we inoculate agar and make transfers u ntill the culture is clean enough to use as innoculant. But BRF is a little more forgiving and will usually do fine from spore syringe.
OP I think you need a PC. Makes sterilizing a breeze. A presto 23qt is a great starter PC holss 10 quart jars at a time, theyre 60-70 bux.
Of course with a PC you could move onto grains and bulk growing and agar work too!
--------------------
 Amateur Mycologists United AMU Q&A
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Machiavelliavore
Vermiculite Hater



Registered: 12/08/14
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Re: Contaminated, again; Six out of Six [Re: mushpunx]
#22450861 - 10/29/15 08:05 PM (8 years, 3 months ago) |
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I've found grains to be more forgiving than cakes. Probably cause I was using rye cakes which are a bit shitty in terms of texture IMO.
Mycellium seems to be very happy in grains, plenty of space to grow through, not sticky.
--------------------
I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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YaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
Loc: NY
Last seen: 5 months, 11 days
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Quote:
microwave methods
You're right, I need that damn pressure cooker first. There's one jar outside that seems to not smell, but it hasn't shown signs of myc yet either
So it's agreed, OP needs a fucking PC
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Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
#22451348 - 10/29/15 09:34 PM (8 years, 3 months ago) |
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Quote:
YaMoonSun said: The syringe making I looked up on a YouTube video
Post the link to this. I've seen alot of terrible syringe tek videos on youtube that have huge flaws.
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