Author Topic: Please help me identify this organism.  (Read 1402 times)

Offline enso

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Please help me identify this organism.
« on: February 13, 2011, 08:07:58 AM »
I realize I won't be able to determine exactly what it is, but ballpark would be nice.  I am hoping someone will recognize the characteristics and appearance.  

A brief history:

A couple of years ago I made up a batch of my heather ale.  This time I decided to add some dry heather to the ferementor at the end of fermentation.  In essence "dry heathering"  ;)

Well, in a few days it appeared to be fermenting again.  I had a bit of a krausen along wit all the heather blossoms floating at the top.  Eventually I kegged it.  It started getting funky.  I debated dumping it many times.  Eventually it started to develop some definite pineapple aromas (eventually I came to the conclusion it was a strain of brett...?) and a bit of sourness.  Many times I opened the keg and started to prepare to dump it, but something told me to keep it.  Eventually I bottled what was left from the keg.  Since that time I have become more interested in wild and sour beers to the point where I realized my infected batch was actually a good thing and I want to recreate it somehow.  I had 2 bottles left.  

About 1 week ago I poured the dregs into a small vial of sterilized starter wort and built it up from there whatever it is.  In the beginning it had the pineapple aromas somewhat but it developed into a much more earthy (fresh wet dirt) aromas.  I also streaked a plate for kicks to see what would grow.

I will post the pics in a bit.  I have to find a host that will let me link here.  Flickr doesn't seem to work.

Any of this sound familiar to anyone?
« Last Edit: February 13, 2011, 08:15:39 AM by enso »
Dave Brush

Offline enso

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Re: Please help me identify this organism.
« Reply #1 on: February 13, 2011, 11:11:06 AM »
Okay, I have a slow connection so it taking a while to upload photos.

Here are some shots of the plate I streaked from the starter.  I hope...

Edit: This is mold.  Still trying to culture the organism described above.  Will post updates




« Last Edit: February 15, 2011, 09:28:34 AM by enso »
Dave Brush

Offline hokerer

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Re: Please help me identify this organism.
« Reply #2 on: February 13, 2011, 01:59:32 PM »
Okay, well I tried google picassa but it is not loading the picture on my connection, so I have no idea if it showing up.    :-[

Is this the one you're trying to show?...


Joe

Offline enso

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Re: Please help me identify this organism.
« Reply #3 on: February 13, 2011, 02:02:38 PM »
Google seems to be having issues, at least for me.  I can't seem to access anything related to google, however other sites seem to work okay...

I am thinking what I have plated is mold.  If I am correct in my understanding, wild yeasts and bacteria are harder to grow on plain malt/agar solutions.  It was not looking like mold at first but I know see some green and it is getting fuzzy.  I am not sure what is in the starter solution.  Perhaps I was not successful in culturing the dregs...
Dave Brush

Offline enso

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Re: Please help me identify this organism.
« Reply #4 on: February 13, 2011, 02:29:24 PM »
Okay, well I tried google picassa but it is not loading the picture on my connection, so I have no idea if it showing up.    :-[

Is this the one you're trying to show?...



Yes that is it.  Thanks Joe.
Dave Brush

Offline enso

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Re: Please help me identify this organism.
« Reply #5 on: February 13, 2011, 02:36:18 PM »
Okay, so without further ado...

The starter.





I set it in the fridge to see if it would settle out.  So far this is all it has done.  Very fluffy.  It had a krausen in the beginning and left some dirty sludge on the jar.  After I added a second half liter of wort it no longer had any kraeusen and that is when it started smelling more earthy.

I am wondering if perhaps I should have attempted to culture whatever it is in beer itself rather than wort.  Thinking that perhaps the lower pH and the presence of alcohol would better recreate the conditions in which I originally obtained this critter.  I am also concurrently attempting to make a mini batch with wyeast 1728 and heather blossoms simultaneously to see if it happens again.  Unfortunately it is not the same heather as the first time.  The first time I had dried heather I bought.  This time I am using some heather that I grew and dried.  It has been in the freezer for over a year...
« Last Edit: February 13, 2011, 02:42:00 PM by enso »
Dave Brush

Offline tschmidlin

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Re: Please help me identify this organism.
« Reply #6 on: February 13, 2011, 03:48:35 PM »
The plates look a lot more like mold than yeast to me.  I've never seen yeast look anything like that on a plate.
Tom Schmidlin

Offline punatic

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Re: Please help me identify this organism.
« Reply #7 on: February 13, 2011, 04:18:59 PM »
I found this picture here:
A 6 - 7 day - old Trichoderma viride mold culture grown on malt extract agar

It looks like it could be a more mature version of your culture.

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Offline enso

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Re: Please help me identify this organism.
« Reply #8 on: February 13, 2011, 08:16:16 PM »
I agree, I now believe it is mold on the plate.  The starter itself I am not sure what I have.  Again, I think whatever it is was too slow to grow on the plate before the mold took hold.  I will try and streak a few more plates and see what I get.  Hopefully I will have a sterile plate the next time.

I am certain whatever "infected" the batch of heather ale was not a mold.  I just may not be able to propagate it.  I have one bottle left...

Do the starter pictures look like mold in the wort?  Generally I think of mold as having some growth on the top of the beer, floating.  This did not have that.  Although I had it on the stirplate for a while.  Perhaps it just settled out when I took it off?

Again, i may attempt to inoculate some beer rather than wort to see if that has better effect.

On another front, the heather infused wort with Wyeast 1728 seems to be doing well.  It is only about 1 pint.  It is definitely fermenting and smells mostly floral and heather like so far.  I added the heather a few days before I pitched the yeast.  One way or another I hope to recreate this accident!
Dave Brush

Offline roffenburger

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Re: Please help me identify this organism.
« Reply #9 on: February 13, 2011, 09:07:14 PM »
What was your procedure for the starter wort. I made some all grain starter wort and sterilized in a pressure cooker and it looked very similar to yours. I am assuming its hot break. Bacterial contamination will generally make your wort cloudy with a few exceptions.

From your pics, I would say that its break material.

When you make your plates, its good practice to leave them at room temperature for about a week. This serves as a sterility check. If your plates are contaminated to begin with, they should show growth within the first week at room temp.
Travis R.

Offline tschmidlin

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Re: Please help me identify this organism.
« Reply #10 on: February 13, 2011, 11:41:28 PM »
It's hard to say what it is in the starter, it could be a flocculent yeast and those are just big chunks, I've seen similar stuff with some brewing yeast.  The low pH and ethanol could be suppressing the mold, so maybe a liquid culture is the way to go. 

Was that really streaked?  Because the growth pattern doesn't seem to reflect streaking.  It's possible the plates are just bad like someone mentioned above.  If not, dilute it before you streak it so you get fewer cells on there, that should make it easier to isolate individual colonies.  Watch it like a hawk, and if you see something that looks like yeast pick the colonies and re-streak to their own fresh plates.  You might get lucky.
Tom Schmidlin

Offline enso

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Re: Please help me identify this organism.
« Reply #11 on: February 14, 2011, 06:59:18 AM »
Pretty sure it is not hot break.  I did make a small all grain batch for the starter.  I let the hot break settle out though.  I cooked it in a pressure cooker and yes, that did result in additional break material.  I then let that settle and filled my mason jars and sterilized them in the pressure cooker.  So, the starter wort was pretty clear t begin with.

I agree, the plate does not look like growth from streaking.  In fact if I am remembering correctly the first streak was actually on the opposite side of the mass of growth.  I remember accidentally cutting the agar on that side (bit soft) when I made the first pass and I can see a cut on that side.

I have been rereading Wild Brews and Chris Whites Yeast book and discovered that plain malt agar is not the ideal media to favor wild yeasts or bacteria, so, I may not be able to grow whatever it is on a plate anyway without the proper growth media.

This is fun.  May not work out but I feel like I am learning a bunch.   :o
Dave Brush

Offline enso

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Re: Please help me identify this organism.
« Reply #12 on: February 15, 2011, 09:44:22 AM »
I restreaked 2 plates yesterday from the starter.  When I dipped the inoculation loop in it came out covered in brown slimey goop.  I am doubtful it will grow anything on the plate.

I have since read some more information.  I believe I may have stepped the bottle dregs up too quickly and allowed some other contaminant (possibly mold) to overtake the starter.  I will have to do some more research before another attempt as I only have one bottle left.   :'(

Anyway, I have not completely given up on this attempt yet.  I have drained off the separated wort seen in the photos above and diluted the gunk in the bottom with a liter of sterilized distilled water.  I then put about 200 ml of some Scottish 60/- I have on tap (2.6 ABV) in a pint mason jar and added ~50 ml of the "gunk" (I guess I will refer to it as such for now...   ::) ) to the jar.

In another related experiment I also added a few grams of heather blossoms to a similar amount of 60/- again in a mason jar to see if I can infect an already brewed beer and get a desirable result.
Dave Brush

Offline enso

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Re: Please help me identify this organism.
« Reply #13 on: February 26, 2011, 01:24:06 PM »
I set this aside for a bit.  I needed my temp controller for some saisions I brewed.  Of the 2 plates I streaked the first was again moldy.  I am fairly confident that what I had in the starter was mold.  It smells like mold.  Here is what a nice cloud of mold floating looks like:



I am confident that the 2 new plates were sterile to begin with.  The plates sat out for over a week @ ~70F after I poured them and nothing had grown before streaking.  The second plate I streaked from from the first without dipping back into the starter.  I left this one in the makeshift incubator (cooler) without heat (again I needed the heating pads for my saisons) for about a week.  It had a small round growth when I left it.  A week later the small growth had developed into what seems to be a lovely mold flower.  I am guessing that is what it is because it appears to be furry.  Seems different from the other mold in that it grew in a floral shape?  I had given up hope on this culturing, however, I noticed some light brown tiny colonies on the plate as seen below circled in red.



I am not sure what these are but they resemble something more akin to yeast growth.  I have added a few of the colonies to a few ml of sterile wort to see what grows.  On another front, my experiment with adding heather blossoms and yeast to some wort is smelling and tasting promising, so I did up a five gallon batch which is fermenting away.
Dave Brush

Offline tschmidlin

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Re: Please help me identify this organism.
« Reply #14 on: February 26, 2011, 09:44:45 PM »
Let us know what you figure out.  Those little brown colonies look more like bacteria than mold to me, but it is really hard to say.  I've never seen yeast colonies that color under normal growing conditions.
Tom Schmidlin