Nectarine Blossom Yeast Isolation

Discussion in 'Homebrewing' started by jbakajust1, Jun 8, 2017.

  1. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    Put a few Nectarine blossoms into 50ml starter vial a few weeks ago. Added slurry to another 50ml vial for step 2. Then plated from the vial. Ended up with some bacteria, yeast, and mold. Took 3 individual colonies each to 3 new plates. No mold on any of the 3 new plates. Took a single colony from each plate and put into 3 new 50ml starters. Swiped a double colony from one plate to look at on a slide. Looks like uniform cells at 250x.

    Plan is to build up from here, run fermentation tests for attenuation and flavor profile. Then choose the best for a full batch.

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    Initial plating at day 4
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    Individual plating of single isolate from plate one.
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    The three new plates
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    250x
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    Initial plating at day 8
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    New vials of individual isolates
     
    #1 jbakajust1, Jun 8, 2017
    Last edited: Jun 8, 2017
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  2. GormBrewhouse

    GormBrewhouse Aspirant (256) Jun 24, 2015 Vermont

    Cool, question, how do you know the bacteria is not harmful to humans? I am guessing there is a way to tell.
     
  3. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    No way for me to tell at home. My buddy has a lab at the brewery and I am going to streak the bacteria to see what grows and take it to him for better identification. Hoping it is a wild Lacto.

    The bacteria formed mostly in quadrents 1 & 2, while the yeast formed mostly in 2 & 3. I took 1 isolate from 2 and 2 from 3. I stayed away from the bacteria as best I could. I don't see any bacteria colonies on the 3 new plates. Any pathogen harmful to humans shouldn't make it through alcoholic fermentation. Need to get a little closer look in the scope but it doesn't look like I see any bacteria cells in with the yeast cells, but 250x won't get close enough for a definitive result.
     
  4. EvenMoreJesus

    EvenMoreJesus Initiate (186) Jun 8, 2017 Pennsylvania

    Morphology/microscopic examination is one way to get a broad idea of what you have, but unless you spend a great deal of time looking into a microscope DNA sequencing is the only way to properly do that.

    The good thing is that most microbes that are involved with fermentation are facultative anaerobes, which tend to be very safe. Couple that with fermentation happening in an environment without oxygen and it's even safer.
     
  5. EvenMoreJesus

    EvenMoreJesus Initiate (186) Jun 8, 2017 Pennsylvania

    This bit's not 100% accurate, but it IS pretty close.
     
  6. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    That's why I said "shouldn't". Always need to be careful when building up wild caught.

    Working on DNA sequencing for a future step...
     
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  7. EvenMoreJesus

    EvenMoreJesus Initiate (186) Jun 8, 2017 Pennsylvania

    As long as you have signs of active fermentation, it smells good, and you've got no visible mold, it should be OK.

    Never had a problem with wild captures in the past, but there's always a chance. There are some VERY good links to the problematic microbes. I can post them, if you like, but I sense that you already know.
     
  8. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    Feel free, others may like the info if they decide to venture into isolating.

    I shake my vials to let out CO2 as they grow. The original step smelled like vanilla. We will see what comes of it in these future steps.
     
    EvenMoreJesus likes this.
  9. GormBrewhouse

    GormBrewhouse Aspirant (256) Jun 24, 2015 Vermont

    Thanks to all for the replys. I always wondered how one could tell good from bad bacteria.
     
  10. hoptualBrew

    hoptualBrew Defender (609) May 29, 2011 Florida
    Beer Trader

    @EvenMoreJesus , welcome to the forums. You're posts and knowledge sharing are great! Cheers
     
    EvenMoreJesus likes this.
  11. DrMindbender

    DrMindbender Champion (893) Jul 13, 2014 South Carolina
    Beer Trader

    Carolina Baeurnhaus's house strain was a wild isolate from Nectarines from upstate SC. They love the yeast and it ferments like a beast! I can taste a little nectarine in many of their beers as well.
     
  12. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    It is really interesting how that happens. My first venture into this type of yeast wrangling was blackberry and peach yeast skin starters that were added to my first real sour beer. After 2.5 years it was judged in a BJCP comp and peach and berry were descriptors. I rebuilt the dregs and fermented a Belgian Blonde ale that had a lot of peachiness to it as well.
     
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  13. DrMindbender

    DrMindbender Champion (893) Jul 13, 2014 South Carolina
    Beer Trader

    I've isolated yeast from local wild blackberries that I use for making meads...always throws a little blackberry nuance in the final product! Carolina Baeurnhaus/SouthYeast also uses a strain from local blueberries that adds just a hint of blueberry that's perfect for a saison! I think they have most of their yeasts available to homebrewers right now if anyone wants to give them a try.
     
  14. Lukass

    Lukass Champion (899) Dec 16, 2012 Ohio
    Beer Trader

    Care to do a yeast trade? :wink: I took your advice last year and harvested from peach skins, and the beer turned out fantastic. It was a blonde ale base. Keep us posted on what you end up brewing, this looks like a fun project
     
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  15. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    For sure. I'm getting ready to step the 3 isolates up to 350ml, hopefully tonight, and take a gravity sample before and after to see what kind of attenuation I get. Going slow on these since I have 3 isolates and 2 stir plates. I'll then bump them up to 2L batches for flavor/aroma/mouthfeel/etc.
     
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  16. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    Last night I did a double pressure cooker run on 400ml of 1.020 wort for the next step. Pitched the three vials this morning into their own starters. Will watch the fermentation for a week or so then give it a taste when I step it up to roughly 1500-1800ml fermentation in a lightly hopped wort. That step will get attenuation tested as well as carbonated and tasted for flavor and aroma. The spent starter wort from the vials didn't give off a distinct aroma, mildly funky, nothing off putting.

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  17. EvenMoreJesus

    EvenMoreJesus Initiate (186) Jun 8, 2017 Pennsylvania

    How much hops? IBUs?

    What are you wanting to inhibit, if anything?
     
  18. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    Probably around 15 IBUs and 1.040 OG. Not really trying to inhibit anything. Want to see how the yeast isolates do with a beer as opposed to simply starter wort.
     
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  19. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    The starters have progressed nicely! Good active fermentation, nice krausen, and clear flocculation. Will step up to 1500ml hopped wort for flavor/aroma/attenuation tonight.

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  20. NiceFly

    NiceFly Aspirant (275) Dec 22, 2011 Tajikistan

    Cool project.

    In the future if you do not want to isolate bacteria you can add an inhibitor of bacterial growth to the agar.

    Ampicillin, Kanamycin and Chloroamphenolcol (spellling?) come to mind. They are usually cheap and a little goes a long way.
     
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  21. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    Thanks. I actually pulled some super small colonies from quadrant one and streaked a new plate. I've got some bacteria in there and also some Brett (I believe, has the umbient nipple). Going to grow them up and see what I get. I plan to gram stain the bacteria and see what I have. I want to grow both up, if I can get the wild Sacc that is already fermenting away, and a viable LAB and Brett it will be super fun to play with a blend.
     
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  22. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    Update: I had stepped up the single isolate starters to 1200ml fermentations for evaluating attenuation, aroma, flavor, mouthfeel. I started with a 1.049 OG, Pils DME, and 12 IBUs from Magnum. I kept it at 65*F for 5 days. Had nice krausen, thick yeast slurry at the bottom of the growlers, and began to drop clear on day 4. Day 6 I cold crashed. Day 7 (last night) I pulled tasting and gravity samples from each of the 3. They all had varying degrees of a cidery note, and pear, and banana. One had a tartness but was very cidery and finished at 73.5% app att. One had a semi-full rounded mouthfeel at 77.6% app att. The stand out was the #2 isolate: Pear, Banana, low cider, low phenols, dry finish, light to medium body, 81.6% app att. I plan to brew up a 12 gallon batch with the #2 isolate on vacation in a couple weeks and use 3 fermenters at different temps to see how it responds to higher and lower temps. I'd say I got a good culture going thus far. (My torch shattered the top of the growler, oops!)
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    #23 jbakajust1, Jun 27, 2017
    Last edited: Jun 27, 2017
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  23. GreenKrusty101

    GreenKrusty101 Defender (675) Dec 4, 2008 Nevada

    Just a thought, but has anyone tried to cultivate bugs growing amongst hops? The cones themselves would likely inhibit most everything...but what about other parts of the hop bine?
    Just musing
     
  24. jbakajust1

    jbakajust1 Moderator (1,062) Aug 25, 2009 Oregon
    Subscriber

    I posted this on the MTF FB page and one response was from someone that isolated from hop cones. I'm going to try it in a couple months when hops are in full swing.
     
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  25. EvenMoreJesus

    EvenMoreJesus Initiate (186) Jun 8, 2017 Pennsylvania

    Just about any fruiting or flowering plant is going to give you something. Whether it's useful or not is the bit that is in question. Most things that are outside and away from heavy human intervention are good bets.

    Anxious to hear what character you get from the culture if you do end up doing the capture.
     
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