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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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stupid amounts of bacterial contam
    #1128132 -

My friend has been having serious amounts of bacterial contams in his jars lately. On avg. 2.5 out of 7 jars contam each batch. He thought it was because he was using the same jars for over a year but even when he bought a new box he still had a contam problem. This is with different grains (rye and finch seed). Where do you think this problem could be comming from? He was thinking maybe it's coming from the isolate but then wouldn't all the jars contam? Should he start fresh with new jars and a new isolate? What is the best way to deal with bacterial problems?


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-SumGuy :cool:

Edited by SumGuy (12/10/02 08:08 AM)

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1128150 -

How do you prepare the grain?


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: stupid amounts of bacterial contam [Re: Anno]
    #1128287 -

Well mostly he's tried 3 different things.

a) 240ml of water and 240ml of rye in a jar and then PC at 15psi for 1hr 15min. (origianl recipie that works pretty good) but he found occasionally he'd get bacterial contams and the grain would be too sticky.

He then moved on to plan B looking for something that doesn't collect as much water.

b) 320ml finch seed to 160ml water - this worked well with the grains not being too sticky and not too wet but still had 2 jars out of every 7 contam with bacteria.

so he wanted to try something a little different that may make all the difference. so he tried plan c.

c) Preboil rye grain for 45 mins or so until they've absorbed the water they need, strain, jar, and PC. This lookd very promising, the grains were very loose and the mycelium liked it a lot for a few days but the incubator seems to be posing a problem with this. The water from the grain evaps. in the jar and collects on the walls causing a reflux type thing to be happening where water droplets would run down the side of the jar and catch on the grain.

He has only been using an incubator for the last two methods because of the cold winter.

Losing one or two jars isn't too much of a problem but he's still angry because the only type of contam he has ever gotten is bacterial so if he could eliminate this problem he could be very successful. The results he gets when the good jars spawned to poo are very very pleasing.


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-SumGuy :cool:

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1128483 -

Try soaking the grain overnight before you boil it on the stove top for 45 min, the overnight soaking lets the grain expand and all of the endo(sp?) spore germinate so that when you PC it they are ALL killed. I think that it also coaxes out any type of dormant contams so that they may be killed more easily.

My buddy does his jars like this (soak, cook, PC-15 p.s.i. 1 1/2 hrs) and has only gotten 2 contams in 10+ batches of jars.

Hope this helps
Dave

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1128658 -

Cook your grain for 90 minutes at 18-20 psi.

add 5 ml of 3% peroxide per qt jar of grain when it has cooled from being p cooked. Shake it up and let it sit for an hour then innoc it with your isolate.

Good luck!

Edited by Millet (12/10/02 11:33 AM)

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1130613 -

how do you stack your jars in the pc? just a thought....


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hmmm........

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1131338 -

i find i get more bacerial contams this time of year for some reason. I think it may be that the home environment is just more closed up and kinda like a massive incubator in itself. I also get more bacterial contams if my incubation temps are on the higher side. I notice also that I get much less bacterial contams if I incubate using temps less than ideal for myc growth. It takes a bit longer but for less contams......


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Support bacteria - they're the only culture some people have.

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1131395 -

One rule of thumb I've found to be pretty good is that bacterial contamination by itself is usually a result of incomplete sterilization. . .not environmental.  Aerial contamination can include bacteria, but if it's screwing up that many jars, you'd expect [statistically] to see a lot more mold problems as well.  If it's just bacteria, odds are sterilization is not going to completion. 
Not a hard and fast rule, but generally seems to be the case. 

A possible exception to this rule is the 'nutrient wick effect', where the sludge from shaken grain forms a nutrient film up to the neck of the jar, allowing bacteria to migrate inside----mold won't do this as readily.  Bacillus is a very motile bacterium, it swims around like crazy when fed well.

If it's giving you significant problems, and the various advice here is a bit overwhelming, devote one batch of jars as 'experimental'.  Vary the water content/preparation technique from one jar to the next, keeping a few notes as to what's what.  After completion, you should have one or two that are 'perfect'. . .pitch the others and make a fresh batch using only that recipe. 

Grain prep is an individual thing, the methods I find easiest don't necessarily work well for others.  Find your groove :smile: 

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Re: stupid amounts of bacterial contam [Re: Suntzu]
    #1131433 -

Stunzu, I think that the 'nutrient wick effect' you're speaking of would most likely be the problem. My friend will try your idea. It's such a bugger to be fairly successful and only having one problem to eliminate. It almost made my friend want to go back a few years to use cakes for spawn hahaha.  :tongue:

Thanks a lot for all your thoughts and suggestions.

DIE BACILLUS DIE! 


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-SumGuy :cool:

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Re: stupid amounts of bacterial contam [Re: upupup]
    #1131878 -

i confirm, i get the highest contam rate from sept trough december


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comario


"crusaders against emotional poverty"

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1132692 -

I feel your pain. I gave up on rye because of all of the bacterial blooms I experienced. I now use millet and have noticed a 10x increase in mold contamination over the last week or two. It looks like I will have to re-evaluate my asceptic technique as well. I think I will rekindle my relationship with peroxide.

Joshua


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The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: stupid amounts of bacterial contam [Re: Joshua]
    #1133166 -

joshua, even the millet was giving my friend freekin bacterial problems... maybe his PC and jars just need a damn good alcohol bath.


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-SumGuy :cool:

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1133263 -

Once bacteria form spores they can be very persistant in the environment.

Joshua


--------------------
The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1133716 -

Does you pc work properly? Does the pressure build up
Did you try this test?
http://www.shroomery.org/forums/showflat.php?Cat=&Board=Forum2&Number=1109176


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Grow Mushrooms from Spores - PF-Tek for Simple Minds

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Re: stupid amounts of bacterial contam [Re: Anno]
    #1134215 -

Anno .. Interesting, he's seen water drip down from the steam collected by the vent and it's never boiled like that... infact it doesn't really boil too much at all it does evap within 30-60 seconds though.. you could be very very right on the PC not working propery...

This is the PC being used, anyone have any comments on it?

http://store.yahoo.com/everythingbagel/presprescan2.html


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-SumGuy :cool:

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1134247 -

I was the queen of bacterial contams about a month ago, but I got a tip from someone here that has made all the difference. I measure my seed, pour a bunch of h202 in with it (enough to cover the seed/grain), cover with tinfoil, and let it sit until it stops bubbling (hours). Then I rinse well, simmer on the stovetop, drain, rinse, drain and pack into jars. My problems have GREATLY decreased - nearly all jars are looking good now. The extra step is a pain in the ass, but not nearly as bad as cleaning stinky jars that have gone bad!


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I don't really know anyone who grows mushrooms. I just come here for fun.

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1134769 -

If you use your PC you shouldn't have alot of contamination problems that's if you use those poly-disks.  Those work pretty good.  Chances are that you introduce bacteria/mold in your jars during innoculation of spores.  When you PC, molds/bacteria don't stand a chance.  I never had a contamination problem in my jars yet.  Here's what I do: i bring water in my pc to boiling point.  Turn off the hit and dump Penningtons' finch in it.  I leave that alone for 30 minutes,  stiring occasionally though not needed.  Then i dump some of it in colandar and wash it in cold water.  *put a little bit in the colander so that you wash off more endospores + most of the water dripps out.  Then if you use your quart jars load it half way with the seed  + I put about a third of wate from the bottom to 1/2 of the jar (seed top*) -/+ 7 cc's :smile: 
I use poly disks those wonders.  Let me make an analogy for bacteria/mold  to get in your jar with out a disk is like throwing a rock at lake that is 20 feet away and you get 100 tries.  But with a disk on u attempting to hit a small puddle 70 feet away & you get one try. 
Now, you start counting when PC pressure is at 15 psi.  I usually PC for 45 minutes oh high - mid high.  Another way of of doing is starting on high & moving down  mid range for 45 mins -55 minutes.  When you turn your PC  off as soon as pressure drops take your jars out and shake em, while hitting it on something soft :smile: so that you break up your millet.  It is very important if you want to speed up your jar colonization.  I have one of those hepa filters from sears, my model outputs fresh air toward the ceilling ( *forget oven-tek ).  It's very convinient to inoculate your jars on it.  And yeah don't forget to rub some alchohol on your foil when you put it back on your jar after innoculation and use a lighter on your syrange.  If you do all that chances are you'll  reduce your contam. rate. 
p.s. use Anno's water drop tek  to check your PC. 



 

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Re: stupid amounts of bacterial contam [Re: SumGuy]
    #1135614 -

Sumguy; I also have that Presto pressure cooker, and am also having more problems than I have in the past. However, I have seen teks written with pictures that show that very unit, so I think it is okay? Not sure, wish someone knew about this. I know that the Presto p/c is very cheap for how large it is, so I have been worried to say the least.

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Re: stupid amounts of bacterial contam [Re: babyshroom]
    #1135620 -

Funny, I heard that using hydrogen peroxide was totally pointless as a pre-soak prior to pressure cooking. I honestly thought that I was having better luck using it just like you, but you could say I was convinced otherwise in this thread (not very long ago at all)
"Some words on effective sterilization" thread

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Re: stupid amounts of bacterial contam [Re: Pugslee_Atoms420]
    #1136298 -

peroxide is pointless in the presoak. The increase in sterility is attributable to some other source. The peroxidiase naturally found in the seed will destroy the peroxide well before it has any sanitizing effect on the inherrent bacteria population, especially the inherent endospores.

That said, the original post should consider using a little less water in his jars. Put the grain and water in the jar, and let them soak overnight. This allows the bacterial endospores (the ones so resistant to heat sterlization) to hatch and become susceptible to sterilization. Simply shake, the sterilize as normal. Assuming that the pc is working, this should show a notable increase in the rate of success.

This is common and is attributable to the source of grain in most cases. The inherrent spore load or bacteria load in a grain usually increases over time. Depending on your source, depends on what time of year you see this, and also depending on how and how well you store your own grains. My bet is that either your source for grains is using some that have been stored for some time at this time of the year, or you are using grains that you have personally had stored for a while. Very common. Oft overlooked.

Do the presoak, and you are using a little too much water anyway.


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"From a certain point of view"
-Jedi Master Obi Wan Kenobi

PM me with any cultivation questions.

I just looked at my profile and realized I had a website at one point in time on geocities, it's not there anymore and I have no idea what I had on it. Anybody remember my website from several years aga? PM if so please.

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