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Offlinepsychotic777
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Registered: 08/16/13
Posts: 20
Last seen: 9 years, 1 month
First timer, having hard time with contams
    #18807968 - 09/06/13 04:06 PM (10 years, 5 months ago)

Hi all,

So hypothetically lets say I'm a first timer at this. I've been having an insanely hard time with contams. I don't know if it's because my apartment that I'm renting is already mold-prone from previous renters or if it's because I live in a tropical South Florida climate.

My first 8 jars were all failures, but that's because I tried doing all of them in the kitchen (dumb) without a glovebox (more dumb). While dealing with my failures I discovered this site and for my next batch I used sponsors for everything. I also built a glovebox and used the genius trick of coffee cans to make the arm holes.

Since I've had so many problems with contams, and money is not an issue for me (within reason), I decided that I would only use stuff that was pre-sterilized from a sponsor to reduce the likelihood that any problems are my own doing. I got 4 quart rye jars and 1 grow rye bag from a sponsor just to see which one I would have the most success with. My biggest priority was getting jars and bag with self-healing injectors because I'm seriously pissed about contams.

Well, as for the actual inoculation process, I made so many dumb mistakes. The first jar I didn't realize which part was the injection port and which was the filter so I tried shoving the needle down the filter. Figured I was doing something wrong when it wouldn't go through so I went back and looked online and realized the injection port is the flat thing. So first jar is already probably going to be screwed up. While going between the 2nd and 3rd jar I realized I had forgotten to heat the needle inbetween jars so I figured another jar was messed up. And by the last jar I realized I didn't use an alcohol swab on the injection port for most of the jars except for the last one.  So really only 1 jar and the bag were done properly, the first 3 jars were iffy or just wrong.
I was so paranoid about contams that I decided to just leave them in the glovebox and put a blanket over it instead of moving them to a top shelf in a closet or something.

I injected 1CC to 1 1/2CC in each jar and about 3CC in the bag.

Anyways this was all done about a week and a half ago. One jar I had to throw out on the 3rd day because I saw green coloring in the jar. Another 2 jars seemed to have growth but also seemed to be drying out so I decided to pull them out and move them to two different closets and observe their growth.  1 jar looks really healthy and I feel like crying in happiness that I at least got one right.  And the rye bag shows absolutely NO growth. Oh yea and I've shaken the bag and shaken the dried out jars a lot to see if it changes anything. Maybe I shouldn't have done that? I dunno.

Attached are pictures of the 3 jars I have and the no growth rye bag. Let me know if you see anything wrong with the jar that I think is healthy, and if I should toss the other 2 jars that seem to be drying out. Also let me know what I should do about the bag.

All pics were taken today 09/06/13, inoculated on 08/28/13

Rye Bag No Growth:






Jar I'm hoping is healthy:





Jar dried out maybe contam #1?





Jar dried out maybe contam #2?




Edited by psychotic777 (09/16/13 10:38 PM)


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Offlinepsychotic777
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Registered: 08/16/13
Posts: 20
Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: psychotic777]
    #18811241 - 09/07/13 11:26 AM (10 years, 5 months ago)

rawr! help me!


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Offlinekrypto2000
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Registered: 12/05/06
Posts: 11,579
Last seen: 4 years, 3 months
Re: First timer, having hard time with contams [Re: psychotic777]
    #18811333 - 09/07/13 11:58 AM (10 years, 5 months ago)

That dried out jar is definitely contaminated. I think you already know what your problem is though, sterilitiy, or the lack there of. Flame sterilize your injection needles next time and wipe down every injection port with alcohol. I'm assuming you pc'd the jars ok and have proper water content, but it's hard to tell from pics alone. Also I would just use .5cc next time too, no real use in using anymore than that and you're just introducing more possible contaminant spores as well as competing mycilium which may or may not get along, plus you simply have more solution for future projects. I would also not put the jars in the SAB or a closet, just put them on a shelf in the open air, light and a little air circulation are good for them and will promote growth and help with gas exchange. Good luck man.

Oh, and your bag, I'm not sure what's up with that, it might be bacterial contamination, it might just be you were shooting blanks :shrug:. Squeeze it so the air comes out through the filter patch and give it a smell check. If it smells funky you definitely have bacterial issues, if not you might want to try to try reinoculating it. Not sure if you want to re pressure cook it to be on the safe side as well or if that would mess up the moisture content, but maybe wait for other members opinions there.


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Offlinepsychotic777
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Registered: 08/16/13
Posts: 20
Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: krypto2000]
    #18812012 - 09/07/13 03:42 PM (10 years, 5 months ago)

Sigh about the dried out jars, but I know what I did wrong...

"I'm assuming you pc'd the jars ok and have proper water content, but it's hard to tell from pics alone."

Actually no I bought them premade from a sponsor because I've had so much bad luck so far that I wanted to minimize the risk of me myself messing something else up. I figure once I can get a clean healthy growth with stuff that is premade for me, then I'll start trying to do that part myself too.

The bag I'm gonna wait another week and if nothing then I'll try shooting another CC and see if anything happens. It spells like normal rye when I squeeze some air out the filter and sniff.


And okay I'll pull the healthy jar out of the glovebox.


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Offlinekrypto2000
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Re: First timer, having hard time with contams [Re: psychotic777]
    #18812019 - 09/07/13 03:46 PM (10 years, 5 months ago)

Prepping grains isn't too hard. If you have any jars already prepped left over I'd suggest sacrificing one and play with it to see what you're going for. Note the moisture content/feel and the weight of the grains. Realize that is how it feels after going through the pressure cooker mind you, but it should give you a good idea of what you're going for. I still over or undersaturate my grains sometimes, but there's enough room for error that it generally doesn't matter; so long as you're sterile throughout the process.

edit: If you got the grain already prepped how could it have dried out?


Edited by krypto2000 (09/07/13 03:49 PM)


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Offlinepsychotic777
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Registered: 08/16/13
Posts: 20
Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: krypto2000]
    #18812063 - 09/07/13 04:02 PM (10 years, 5 months ago)

"If you have any jars already prepped left over I'd suggest sacrificing one and play with it to see what you're going for."

Well yea with the rye in the bag I moved it around and could see how it should basically feel. Maybe when I'm ready to prep my own jars I'll do what you suggested. And I'd have to get a pressure cooker to start with... looking on amazon the price range is all over the place.


"edit: If you got the grain already prepped how could it have dried out? "

I'm guessing the bacteria or w/e contam that is in there dried them out after inoculation. They didn't look like that at all when I started.


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Offlinepsychotic777
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Registered: 08/16/13
Posts: 20
Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: psychotic777]
    #18828732 - 09/11/13 04:55 PM (10 years, 5 months ago)

09/11/13

Update. So I tossed out the unhealthy 2 jars. Rye bag still has no growth.



my only happy jar is still colonizing. This last section is taking forever to colonize it seems. How much longer do you all think it will take before it finishes?

Also I have some questions about the next step. I'm using perlite in a large blue plastic bin (light can still shine through the top and shines blue) with an air filter connected to it.  I had soaked the perlite with water and hydrogen peroxide and then strained it all based on the recommendations of another site. Anyways my humidity reader shows that it's still consistently at 99% humidity. Is too much humidity bad? I can use fresh perlite and try not to get it so soaked. I plan on using 3-4 inches of perlite.


Anyways pics:


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OfflineMush4Brains
LOOL HACKED!!!

Registered: 07/31/13
Posts: 4,419
Last seen: 9 years, 3 months
Re: First timer, having hard time with contams [Re: psychotic777]
    #18828754 - 09/11/13 04:58 PM (10 years, 5 months ago)

Have you shaken that jar?  If you haven't, if you do it now that will be done in two days tops, plus, it will help bring out possible contams before you spawn it.


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Offlinepsychotic777
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Registered: 08/16/13
Posts: 20
Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: Mush4Brains]
    #18828767 - 09/11/13 05:00 PM (10 years, 5 months ago)

Okay! Shaking it now!

edit: it broke apart when I shook it, so I hope it's okay :<
also it really looks like it needs more time because a lot of the inner areas didn't have any mycelium.


Edited by psychotic777 (09/11/13 05:04 PM)


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InvisibleHypnotoadCroaked
Retired, but will check MSGs

Registered: 01/05/13
Posts: 1,168
Re: First timer, having hard time with contams [Re: psychotic777]
    #18828815 - 09/11/13 05:10 PM (10 years, 5 months ago)

Keep the bag.  I got into this hobby with a pile of pre-made bags.  I had one bag in particular take well over a month to show any growth at all.  I did a happy dance when it happened.  Give it time.  It takes more effort to throw it in the trash than to let it sit there for a few more weeks.


Quote:

psychotic777 said:
Okay! Shaking it now!

edit: it broke apart when I shook it, so I hope it's okay :<
also it really looks like it needs more time because a lot of the inner areas didn't have any mycelium.



If you injected against the glass as prescribed, you will see this.  As above, be eazy.  These living things move at their pace, not yours.  Put them into "time out" and make yourself only check them once or twice a week right now.  Checking ever few hours only sets you up for continual disappointment.


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InvisibleNice Ol Bud
Apprentice


Registered: 09/11/13
Posts: 162
Loc: Mystical Maze of Mushroom...
Re: First timer, having hard time with contams [Re: HypnotoadCroaked]
    #18828833 - 09/11/13 05:13 PM (10 years, 5 months ago)

Your doing good.
Don't shake it again.
Don't listen to whippy. You can literally watch the mycelium grow on rye.
It's pretty fast man.
Anyways they look great. Keep it going.


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


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Offlinepsychotic777
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Posts: 20
Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: Nice Ol Bud]
    #18828851 - 09/11/13 05:15 PM (10 years, 5 months ago)

Yay :smile: I'm excited.

Ummm nobody really answered my question on the humidity though. Will spawning them in 99% humidity be bad? Or is that fine?


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Offlinemason man
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Registered: 06/01/13
Posts: 69
Last seen: 9 years, 11 months
Re: First timer, having hard time with contams [Re: Mush4Brains]
    #18828871 - 09/11/13 05:17 PM (10 years, 5 months ago)

Quote:

psychotic777 said:
Okay! Shaking it now!

edit: it broke apart when I shook it, so I hope it's okay :<
also it really looks like it needs more time because a lot of the inner areas didn't have any mycelium.




It's alright, after you shake it normally looks like it's 0% colonized. It takes a day or so to recover, but when it does, you'll be surprised at how far along it is. Don't forget to consolidate for a few days after it appears 100%.

Also, I noticed that you took someone's advice on here within one minute of reading it. Not that there is anything wrong with that advice or the person who offered it (I second the shake idea), just be sure to research things on your own instead of just taking what one person says and doing it right away, just to make sure you're getting valid advice. The search feature is everyone's best friend here.

Quote:

Mush4Brains said:
Have you shaken that jar?  If you haven't, if you do it now that will be done in two days tops, plus, it will help bring out possible contams before you spawn it.




Who's the frequent poster on these forums that swears by shaking his jars like 5 times, and colonizes full jars within a few days? I can't remember his name, but I kind of want to read through his posts.


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InvisibleNice Ol Bud
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Registered: 09/11/13
Posts: 162
Loc: Mystical Maze of Mushroom...
Re: First timer, having hard time with contams [Re: psychotic777]
    #18828880 - 09/11/13 05:18 PM (10 years, 5 months ago)

Quote:

psychotic777 said:
Yay :smile: I'm excited.

Ummm nobody really answered my question on the humidity though. Will spawning them in 99% humidity be bad? Or is that fine?




Hell naw that's great :smile:


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


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Offlinepsychotic777
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Registered: 08/16/13
Posts: 20
Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: Nice Ol Bud]
    #18840833 - 09/14/13 11:43 AM (10 years, 5 months ago)

09/14/13

Whelp update. After shaking several days ago on 09/11/13 it hasn't changed. It looks exactly the same way it did after shaking it. Only difference is that the cloudiness at the top of the jar is gone and the mycelium is either all clumped together tightly in different bunches or just scattered around thinly.

I was reading the boards and some people were saying stalled growth could be due to a lack of gas exchange, and they solve it by flipping it upside down and loosening the lid. Thing is this jar has a polyfill filter at the top so I thought that already solves the problem of gas exchange.

So my guess is maybe when I shook it there was some bacteria/contam that has stopped it maybe? Or maybe there is a problem with gas exchange and I should flip it upside down and loosen the lid? or maybe I'm just unlucky and mine is extra super slow? lol. hm..


anyways I have some new jars that I'm going to be inoculating to try again today, but I'll just be leaving the uncolonized bag and this jar on the top shelf to see if anything magically changes in the next week or two.



Edited by psychotic777 (09/14/13 12:33 PM)


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Offlinepsychotic777
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Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: psychotic777]
    #18841652 - 09/14/13 04:59 PM (10 years, 5 months ago)

So yea I just got done inoculating 4 new jars and this time I did it perfectly ^_^ So I have high hopes for these. If something still goes wrong with them, then I'm just going to start boiling the jars before using them even though they are supposed to be sterilized and ready to use right out of the bag (well after using alcohol swab on the injector port + flame needle).

I still don't know what's going on with my previous jar that I shook. Will leave it alone for another week I guess.... Or should I flip it and loosen the lid?


I also had ordered some extra perlite from the sponsor but the perlite is totally different than the one I had ready. The one I had ready were very small and all white, these that came in the mail are much larger and actually have some weird brownish/black stuff in them. It looks like wood or some other rock to me or maybe I'm crazy. I looked up pictures and couldn't find anything that looked like it anywhere so I thought I'd post pictures here and ask what the heck they are? So yea, is this perlite safe to use or should I toss it?






Edited by psychotic777 (09/14/13 05:10 PM)


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OfflineEarthshaker
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Registered: 09/10/13
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Last seen: 3 years, 9 months
Re: First timer, having hard time with contams [Re: psychotic777]
    #18844758 - 09/15/13 02:23 PM (10 years, 5 months ago)

Hi psychotic777,
When I started, I had the same kind of problem like you. It took me about 6 months until I understood what my problem was, and from what I have learned, I could give you the following advices:

  • Generally, I don't agree with spore syringes as a beginner, because it wastes a lot of substrates until you know how to work in a sterile way. I recommend switching to petri dishes and trying to inoculate them with spores, using a scalpel and an alcohol flame. Contaminants are easier and quicker to detect, and you need only one tenth of a square millimeter of spores per petri dish. Once you have mycellium going on a sterile petri dish, you can use this petri dish to inoculate substrate jars and other petri dishes. This means, you need spores only once, and then you work with petri dishes. You can store them in the fridge for a long time (I do it for up to a year). You should get pyrex glass petri dishes, because you can sterilize them over and over again. 10 dishes would be sufficient. Agar comes from the local organic food store :laugh: and you can just boil a couple of potatoes with the skin and use the water plus dextrose to make the agar nutrient media.

  • Your Glove box doesn't need to be fancy, it can be as simple as you want. I just use a frame on which I place some see-through plastic tarp (for painting). Then I spray a 70% isopropanol spray inside and wait until it has settled down (2-3 mins). I spray the same isopropanol on my disposable vinyl gloves and wait 2-3 minutes as well. Gloves sanitized with isoprop are mandatory.

  • In my experience, a sterile environment is not important, as long as you use your vinyl gloves, the alcohol spray and the glovebox properly.

  • Prepare the lids of your jars with polyester filling (I got it from the inside of a pillow). This avoids contaminants to enter the jar while oxygen exchange is granted.

  • And most important of all:
    Don't buy anything from somewhere else. If you can't make everything yourself, your method is not ready yet. It is important that you master every single step, because if you have sterility issues yet, growing fungi can be a big health risk.



I hope my advice is not too devastating. Remember, there is a steep learning curve, and once you master each step, you will suddenly realize how easy it is.


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Offlinepsychotic777
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Registered: 08/16/13
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Last seen: 9 years, 1 month
Re: First timer, having hard time with contams [Re: Earthshaker]
    #18850181 - 09/16/13 05:02 PM (10 years, 5 months ago)

Thanks for the great advice Earthshaker. Honestly you've inspired me that for my next batch I will try doing everything from scratch and not rely on buying anything from anywhere else.

I'm still hesitant about using petri dishes over syringes since I've heard many others argue it's harder, but the rest of that sounds good or more or less what I already have.



09/16/13 - Also as an update, out of nowhere it struck me that part of my problem with having such slow signs of growth (in particular with the rye bag and my healthy jar not showing any changes after shaking) might be due to the temperature of my apartment... I have blackout curtains over all my windows and I keep my place ice cold freezing while I'm home which got me suspecting I might be keeping my place way colder than what most others would consider room temperature :P (as a fyi, blackout curtains are a huge A/C energy cost saver since they keep sunlight and heat out and keep the inside cold).

Anyways I threw my extra humidity/thermometer in the closet where I was incubating all my jars and was shocked to find that it was mostly at 69-70 degrees Fahrenheit with a brief daily high at around 73 (i.e. when I'm not home and give the a/c a break). After doing some research it seems that they should be kept at around 75-80F for ideal growth since anything above that they might overheat (since the jars give off heat and you don't want it over 85)

Using the search tool I found this great homemade incubator:
http://www.fungifun.org/English/Pftek#a5
I already had the perfect two containers ready for it, I used 2 soup cans for the weights on the bottom, and pulled out my fish tank's heater (the fish never use it anyways). Now all my jars are sitting at a happy 77-79 degrees :smile:  I will be keeping a close eye on it, but I suspect this will solve a major problem I've been having with extremely slow to nonexistent growth. Or at least I hope it will!


edit: also I read somewhere else that the weird brown/black perlite stuff is still safe to use, but I'm waiting for more comments on if it's safe or not.


Edited by psychotic777 (09/16/13 05:12 PM)


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InvisiblePestile

Registered: 05/02/13
Posts: 875
Loc: Northern Europe
Re: First timer, having hard time with contams [Re: psychotic777]
    #18850225 - 09/16/13 05:12 PM (10 years, 5 months ago)

You're supposed to shake the big clumps loose. They will stick together with mycelium, just give them a good bashing and spread those colonized grains around. In a couple of days you'll see the effect of the shaking.


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


The Corbett Report
Open Source Intelligence News



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OfflineDeadPhan
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Registered: 05/05/04
Posts: 5,260
Last seen: 4 years, 3 months
Re: First timer, having hard time with contams [Re: Pestile]
    #18851174 - 09/16/13 08:44 PM (10 years, 5 months ago)

What method of making lids did you do?


--------------------
 


Big Gulps!  Alright!  Well,  See ya later!
And if i claim to be a wise man, well, it surely means that i dont know!


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