bacteria contamination and grain to grain.

[Faithjuice]

Hey guys, would appreciate your advice. I have a rye jar that is about 20% colonised with strong my mycelium in one area, it's about right for a shake, however there are small signs of bacterial contamination in noncolonised parts of the jar, there is not a lot of it, but a few grains here and there look slightly jellyfied and weeping a bit.

Do jars have to be perfectly contamination free, or can the mycelium overcome minor bacterial infection? will that bacteria comeback and attacked the substrate?

I have two other jars which also had small signs of contamination, I have shaken a few days ago which seemed to be doing okay.

I am considering salvaging the good mycelium and doing a grain to grain, is that sensible or just a waste of effort. my objective is to get a few good sporeprints

[MLBjammer]

If it's bacterial contamination, it will normally spread like crazy and smell really bad (stinky feet, sour apples). I would give your jar a day or two after the shake and check for any wet spot or bad smell. Sometimes moisture simply pools in the bottom of grain jars, and a shake will redistribute that moisture.

If you do find contamination, you probably don't want to try a grain transfer because that contamination will ride along with seemingly clean colonized grain--and you will just spread the problem.

[Faithjuice]

Originally Posted by MLBjammer

If it's bacterial contamination, it will normally spread like crazy and smell really bad (stinky feet, sour apples). I would give your jar a day or two after the shake and check for any wet spot or bad smell. Sometimes moisture simply pools in the bottom of grain jars, and a shake will redistribute that moisture.

If you do find contamination, you probably don't want to try a grain transfer because that contamination will ride along with seeming clean colonized grain--and you will just spread the problem.

Hi MLBjammer, thanks for your reply, bottom line is my syringe is contaminated, and I've set my mind on overcoming the problem rather than acquiring a new one, I like the idea of keeping my genetic line alive,

I squirted some spores on to a porridge agar substitute, I did some transferring of mycelium in a glovebox to some small jars using peroxide, this was really just a practice run to see what happened, after which I smelt the porridge jar and it had bacteria contamination, the porridge slab was not fully colonised as I was in bit of a hurry.

Anyway I hope these pictures show what I mean, they are all of the same jar, the contamination is at the top of the pictures.

Here are some pics

[MLBjammer]

I would give them a shake. Then you will know what's going on in those jars within a day or two. Sometimes it is hard for me to tell early mycelium from something else. I do not see anything that is obviously bad, and the last two pictures look good.

You can always add a spoon or two of vermiculite in the bottom of your jars to absorb excess moisture.

[BuckarooBanzai]

I don't do rye, but that grain doesn't look great to me. Lots of the kernels are split, though, so I'm not certain if the goo I'm seeing is sugars or bacteria.

Smell is the giveaway - if it stinks, it's bacteria.

It is very difficult to clean up a dirty culture source with grain.

You need some agar. And you don't want the agar to fully colonize.

Inoculate with a few drops of spore water and then transfer the myc within about 2-3 days of seeing it. Myc on agar incubated at 86F will "outrun" bacteria. After 3-5 transfers onto clean agar, you will be left with nothing but healthy mycelia. The transfer cleanup process is extremely reliable, but also extremely tedious and time consuming.

I would consider acquiring a clean print or another syringe.

[red_lenses]

Looks like bacteria to me.
Sometimes if you dont shake it you can let a nice clean area colonize then do a G2G transfer to a clean jar of grain.
Take the myc right off the top though,and don't touch the spoon to any other part of the jar.

Red

[MLBjammer]

I should have read your post more carefully, Faith. It is tough to tell if that jar is skunked. If it is, the bacteria will spread quickly and really, really stink.

I dunno how comfortable you'd be working with agar, but like Buck said, that is the only way to isolate the mycelium from the baddies.

If you need a print to start anew, just PM me.

[Myc]

If the jar is bacterially contaminated
And you shake it
You'll know for sure soon enough

Bacteria will win that battle.

I have some really neat cultures which I've rescued from contaminated jars.
Just do like Buck says,
Agar work.

Give that jar a whiff - it may not be contaminated.
I'm looking hard and what I see could just be sloppy grain - no big deal - just slow things down a bit.

[MLBjammer]

That is what I thought, too, Myc. When I have had jars go bad, I could smell them from several rooms away in my house. And my wife accused me of having smelly tennis shoes . . . I wish that were the case!

[Faithjuice]

Hi, it's rather hard to tell what the jar smell like without opening them, the ones I threw out definitely smelled when I opened the lids but not really before. MLBjammer I've shaken my two other jars, I have some pictures attached, I suppose it's a question of waiting and see. those jars were the first ones I did, and boiled them for too long, after a bit more reading I think I got the technique down. don't like using vermiculite, it makes the jars less pretty.

Forgive my pigheaded, newbe ignorance, but I don't buy it, that it is necessary to use agar. I don't really understand the agar preparation technique, and I am just too dam lazy and too cheap to buy any. What you see in one of the pictures is a strip of mycelium coated porridge, I gave it a peroxide dunk, but my peroxide pooped out of me and stopped fizzing in the glove box. After doing this I smelt my porridge culture and it was funky with bacteria. Everything screams this won't work, but I bet you it does, crazy shit happens, I have six of these little pots and we shall see.

Thanks for your kind offer MLBjammer really appreciate it, but it means a lot to me to revive the genetics that I got from one fresh mushrooms that I bought many years ago, it's almost like family.

[BuckarooBanzai]

Just putting it on agar isn't enough and it doesn't have to be agar. Any flat nutritional surface will work.

The secret is transferring. Cut fast/healthy growth and move to a new jar/flat surface. Do it again. Do it a again. After enough transfers, you will only be moving clean growth. You could do that with grain jars, but it will be more problematic.

Then you inoculate the jars with the clean growth.

A peroxide dip is not going to be sufficient to kill bacterial endospores.

The purpose of using agar is that you can control the nutrients in it and favor fast mycelia growth. If it grows fast, it outruns the contams and a few transfers removes the contams from the culture.

[Faithjuice]

Originally Posted by BuckarooBanzai

Just putting it on agar isn't enough and it doesn't have to be agar. Any flat nutritional surface will work.

The secret is transferring. Cut fast/healthy growth and move to a new jar/flat surface. Do it again. Do it a again. After enough transfers, you will only be moving clean growth. You could do that with grain jars, but it will be more problematic.

Then you inoculate the jars with the clean growth.

A peroxide dip is not going to be sufficient to kill bacterial endospores.

The purpose of using agar is that you can control the nutrients in it and favor fast mycelia growth. If it grows fast, it outruns the contams and a few transfers removes the contams from the culture.

Okay I recant my heretical ideas, thanks for the input, seriously I appreciate it.

[BuckarooBanzai]

No problem, man. I'm just trying to provide constructive input. No need to recant - I'm not preachin at you and you ain't preachin at me. We are just sharing information!

In the end, the grow decisions are yours. I just try to do whatever I can to help folks succeed.

[Myc]

I'll back the "nutritional media" idea
no need to use the word agar here.
Just a two-dimensional surface which allows the easy detection of foreigners

Snag some of your desired cultivar before the contam catches up.
Transfer to another "nutritional medium".

Problem with not using agar is that sometimes the fungal cells become infected. As if you had a cold, flu, herpes simplex virus (cold sore), etc.
I use gent. agar to help them fight off infection upon initial transfer.
Then go to regular media.

Gentamycin can easily be added to any sterile "nutritional medium" prior to pc'ing.
Just use it as a cleanup step and not as a main substrate.

[MLBjammer]

Faith, I guess you don't have filters on your jars. That is how I smell them--polyfill filters on my jar lids.

Good luck.

[Faithjuice]

Originally Posted by BuckarooBanzai

No problem, man. I'm just trying to provide constructive input. No need to recant - I'm not preachin at you and you ain't preachin at me. We are just sharing information!

In the end, the grow decisions are yours. I just try to do whatever I can to help folks succeed.

I was only joking around with the recant remark, I really enjoy reading your comments on the forum in general. MLBjammer yes I have got holes in the lids, just a bit on the small side about 1/16 of an inch. I will let you all know how it goes.

[MLBjammer]

Good luck, man. I hope that you can save your mycelium.

[Faithjuice]

I've found out that I don't seem to have any contamination of any sort, I should have waited and been more patient. The jars I have shaken smell of pure fresh mushroom. Even the porridge slab jars, that I cloned from I think were okay, I was using jars that had pickle in them, and the lids still retained the sour smell, that I thought were bacteria.

I've thrown out eight jars mostly popcorn that I thought were contaminated, but it probably was just that the grain was this rather wet and was leaching carbohydrate and the spores were very old and taking there time. sorry for all the drama.