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Old 01-20-2013, 02:35 AM   #891
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Is it too cold here in south eastern New York to do this right now ? It ranging between 28-38 degrees F.

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Old 01-24-2013, 03:55 PM   #892
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One of the 3 wild Brett I have growing just formed a pellicle.

These are the yeast that formed this pellicle.

5-funk-.jpg   bm-5jpg.jpg  
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Old 01-24-2013, 04:10 PM   #893
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I've captured wild Sacc, but I've only been interested in Brett so I've never kept any strains or tested them out. Based on my wild collections, I seem to get more Sacc (and possibly other non-Bretts) when the temp is warmer outside. Not sure if there is a real correlation since I've only done a handful of collections. It's help if you have some way to make selective plates/media or can put your yeast under a microscope.

I'll ask another question. I have 3 different wild Brett strains fermenting a simple wheat extract. They are about 2.5 months old and in the past week, 2 of them started to get really dark. I'm talking about a light wheat beer to a vary dark amber colored beer. I did a bit of searching and haven't found any answers. Anyone ever seen anything like this? I'll post pictures tomorrow.
I did sample them a few weeks back and maybe the air exposure caused some oxidation or some other reaction? I've never seen this with other beers I've brewed, but this is the first time I've done such small volumes.

Here is a photo. All started like the gallon on the right.
The color difference is not due to lighting, although all three are a bit darker due to the poor quality photo.

Any ideas?
Thanks.
I have seen the opposite, going to very light. Are you sure they are Brett strains? How did you profile them?
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Old 01-24-2013, 04:15 PM   #894
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thanks a lot. very helpful information. i'll try both suggestions (wln media and less drugs). i have been growing liquid cultures at 30 but plates at room temp (trying to be a bit inconspicuous at work) which i guess is ~21.
i haven't seen that 3 week plate suggestion, like you it goes against my microbiology instincts.
unfortunately i'll have to go open a new bottle. girardin or cantillon?? or oud beersel? such choices.
cheers!!
Take it very easy on the antibiotics, the cells are very weak when they are dorment and at the bottom of a bottle for a long time. Sometimes its even better to leave them out completely and go by visual inspection of colonies. 3 weeks is not absurdly long, especially for half-dead yeasts.
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Old 01-24-2013, 04:19 PM   #895
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thanks a lot. very helpful information. i'll try both suggestions (wln media and less drugs). i have been growing liquid cultures at 30 but plates at room temp (trying to be a bit inconspicuous at work) which i guess is ~21.
i haven't seen that 3 week plate suggestion, like you it goes against my microbiology instincts.
unfortunately i'll have to go open a new bottle. girardin or cantillon?? or oud beersel? such choices.
cheers!!
But you are in Utrecht! You should be able to get those bottles pretty cheap and easy!
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Old 01-24-2013, 09:48 PM   #896
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I have seen the opposite, going to very light. Are you sure they are Brett strains? How did you profile them?
Huh, that's interesting.

I putatively identify Sacc (and maybe others?) vs Brett first by collecting everything on an agar plate with either no drugs or just antibiotic to inhibit bacterial growth. Then I streak out individual colonies on a plate with cycloheximide and bromocresol green. The Brett guys are going to be cycloheximide resistant and white to light blue in color.

I then grow them in liquid culture and look at the morphology under a microscope. I've asked a few friends that study Sacc, and they've all said 'definitely not Sacc'. I showed one to a guy that studies Brett and he agreed that it's more than likely a Brett strain.

Most of what I collect seems to be sensitive to cycloheximide and grow blue/dark blue on bromocresol green. More than the opposite. It also seems to be a bit temp and location dependent.

Sure this doesn't definitively identify these guys as Brett, but I think very few other yeast will fall into this profile. Haven't done any PCR yet, but may once I find a strain that makes a good beer that I like.

All that plusthey don't taste like a Sacc beer!

I'm not a yeast geneticist, but this is what I've gathered from talking to yeast people and doing a bit of reading of the literature.
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Old 01-25-2013, 07:45 AM   #897
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Take it very easy on the antibiotics, the cells are very weak when they are dorment and at the bottom of a bottle for a long time. Sometimes its even better to leave them out completely and go by visual inspection of colonies. 3 weeks is not absurdly long, especially for half-dead yeasts.
ok, thanks for that. the last time i plated dregs i had only one plate left and (embarrassingly) no control plate, so i just spread them to see what i got. are chx/chlor even still active after 3 weeks @RT? or is early selection sufficient to weed out bugs/sacc/etc? anyways i'll do it properly next week with low drugs and -drugs.

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But you are in Utrecht! You should be able to get those bottles pretty cheap and easy!
yeah, i know. the things i do for this hobby.... but imagine my dilemma, even as we speak i have kriek or gueuze from 5 different breweries in my fridge... how do i choose??
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Old 01-25-2013, 07:48 AM   #898
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hey bluedubbed which continent of our bluegreen earth do you inhabit?

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Old 01-25-2013, 04:33 PM   #899
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I'm in NA - Utah. We live at about 8000 ft and most of my samplings have been in the mountains between 7-10,000 ft. Probably has something to do with what yeast I'm getting.
I'm not sure about chx, but I use chlor plates (for bacterial work) that are 5-8 months old.
Let us know what happens next time you plate.
Such a dilemma! We only get Lindemans in UT, which is great the first time you try a lambic...
It's always fun going out of state and seeing all the different lambics/sours that are available.

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Old 01-25-2013, 04:44 PM   #900
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I'm in NA - Utah. We live at about 8000 ft and most of my samplings have been in the mountains between 7-10,000 ft. Probably has something to do with what yeast I'm getting.
I'm not sure about chx, but I use chlor plates (for bacterial work) that are 5-8 months old.
Let us know what happens next time you plate.
Such a dilemma! We only get Lindemans in UT, which is great the first time you try a lambic...
It's always fun going out of state and seeing all the different lambics/sours that are available.
I cannot get my hands on chx here and use ampicillin to keep bacteria at bay.
Chloramphenicol is pretty stable, you can even autoclave it I believe, and that is not the case for ampicillin for sure.
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