[gandelf]

Would anyone have a tried and true process for settling lacto in a starter?

[majorvices]

I don't think you need a starter, just pitch a vial or smack pack. Unlike growing yeast for a beer lacto, pedio and the like don't need high cell counts and they don't have to have their cell walls in tact to grow. Just pitch it in and let it do its job.

[gandelf]

I don't think you need a starter, just pitch a vial or smack pack. Unlike growing yeast for a beer lacto, pedio and the like don't need high cell counts and they don't have to have their cell walls in tact to grow. Just pitch it in and let it do its job.

I use to have that position also, until I tried mashweasel's process of utilizing a lacto starter on his Berliner Weiss. I have been doing a sour ale or two/year for the last few years and have fund that making a lacto starter significantly reduces the time required to achieve the desired level of lactic acid I'm looking for. I don't sour mash due to the unreliable flavor profile in my early attempts.

I now mash, boil (no hops) 15 minutes, cool to 100 F, pitch starter with a layer of cling film on the surface of the wort in my kettle. Then do a normal boil when my desired level of sourness has happened and then my ale yeast routine.

I'm content with my sour ales; I just want to reduce the amount of lacto starter wort in my batches.

[Jimmy K]

Lacto pitched without a starter can take a very long time to sour. Doesn't really work for quick styles like berliner weisse. Starters really need to be kept very warm - like 100-110F to grow lacto.

[mabrungard]

I could not find Kris England's (mashweasel) recommendations for Lacto starters, so I don't know what those recommendations are.  (Could someone post the link?)  But, there are some very good Berliner brewers in my club that have given me advice on this subject. 

In the case where you are not using a pure Lacto culture and are doing the handful of raw grain in wort trick...you have to let that starter culture ferment completely in order to let the Lactobacillus out compete and kill off the other organisms with Lacto's superior pH tolerance.  So you let the pH of the starter to get very low and for all the 'off' aromas to dissipate to the singularly lactic aroma.  Of course, you need to ferment the starter at a relatively high temp (near 100F) to enhance the Lacto growth.   This starter should be extremely sour.  Those resulting Lacto bugs are then pitched into the beer batch.  If desired, the partially Lacto fermented beer can be boiled to kill the Lacto and then a regular ale yeast added to complete the ferment.

I love sours! 

[gandelf]

I don't have the link handy Martin, but here is what I have on Kris' comments concerning a lacto starter. I post this with all due respect to the weasel.

Mashweasel
Bacteria/yeast at a 3 parts bacteria to 1 part yeast is the pitch ratio? Did some counts on the yeast and bacteria for you all as follows: Wyeast 1007 10^9/ ml Lacto 10^6 to 10^7/ ml It’s about 100-1000 fold difference between the two. NO WHERE near the 3:1 - 5:1 yeast.

Most people don’t understand this. You will definitely need a starter for the Lacto. NO hops in the starter. I do mine at 37C (100F) shaking overnight and that works fine. If you do it at room temp, it will take longer by far.
Starter gravity - 1025-30
Culture size - 10ml to 100ml to 1L is fine
Stir plate - Y-E-S!!!
Starter and fridge - sure
Incubator - They are VERY easy to 'make'. Cardboard box, whole in the top, surround the whole with foil. ANY bare light bulb type lamp will work. I’ve used of one of those 'car work lights'...the ones with the cages. I have a PID set on mine so it doesn’t get hotter than 98F. However if you are doing the cardboard method you won’t get over 98F. Probably around 90F. Which is fine.
Yeast - Yeah, one tube or smack pack should be fine.
The funny thing is, if you use this method I’ve NEVER had a Berliner Weiss that was sour. Don’t worry about pitching to much Lacto. I repitch lacto AND yeast when I bottle condition at the same ratio 3:1, yeast/lacto.

You shouldn't be worrying about how sour your culture is, just the number of Lacto. L. delbrükii is a homofermentive strain which means its major product is lactate. Sugars (mostly glucose) are metabolized via glycolysis into pyruvate and then into lactate. I use MRS media and agar to grow my lacto for stocks. To start for a batch I'll make a starter that is half glucose and half malt extract to about 1.025 (2oz/qrt). You can go higher if you would like, 1.030-35 is fine also. I'll then put on my shaker at 200rpm at 37C overnight (16h or so). This will create more than enough lacto for you to use. The three key points again are:
1. 50% glucose
2. Stir plate or shaker ~200rpm
3. 37C, probably the most important of the 3
I'll then put them in the fridge, add a little silica-based clarifier and let them drop out. They are much wispier than yeast and don't floc near the same. Decant and add a small amount of starter (malt extract only this time) and let it go for a bit on the stir plate at room temp before you pitch; this way you’ll have plenty of lacto AND they will be ready for beer.

[james]

I'm not sure if there is another place that Kris posted all his awesome berliner info, but I had this post at the NB forum bookmarked.

http://forum.northernbrewer.com/viewtopic.php?f=4&t=43587

[wildknight]

To answer you original question, time and refrigeration is all you can really do to settle a lacto starter.

However...

I decanted a sour pack into 500 ml of wort and put it on a stir plate for 24 hours.  Since the starter wort was simple and the same gravity as my Berlinerweisse, I just pitched the whole thing into the fermenter.  After 24 hours on the stir plate, the starter smelled amazing.  All the sour flavors and aromas that were created in the starter, I definitely wanted everything in my beer.  Remember, bugs replicate every ~30 minutes, whereas yeast replicate every  ~8 hours (on average, of course).  You get plenty of growth with bacteria in just 24 hours. 

I think it is more important to get the bugs ramped up and pitch them cranking than it is to increase the cell counts via the traditional yeast starter methods.

[tschmidlin]

Remember, bugs replicate every ~30 minutes, whereas yeast replicate every  ~8 hours (on average, of course).  You get plenty of growth with bacteria in just 24 hours. 

That holds for e coli and probably some other bugs, but for lactobacillus it's more like 2-3 hours at best (depending on your conditions).  I agree though, lots of growth in 24 hours.

[wildknight]

Remember, bugs replicate every ~30 minutes, whereas yeast replicate every  ~8 hours (on average, of course).  You get plenty of growth with bacteria in just 24 hours. 

That holds for e coli and probably some other bugs, but for lactobacillus it's more like 2-3 hours at best (depending on your conditions).  I agree though, lots of growth in 24 hours.

If you make your bug cultures at blood temperature (37C) or warmer, then you will get closer to the 30-45 minute estimate.  I think someone already posted something relative to these temperatures in their protocol above.  However, if you are simply making a starter in your cool basement, then you are right and you probably will be closer to 2 hours per replication. 

[gandelf]

Thanks Wild Knight and Tom for your informative response to the QUESTION OF THE THREAD. All too often,
I see people responding in an obtuse manner; with the thread dying and going unanswered. I agree with
your statements, having brewed many sour ales from blonde to black. Rather than decanting; I'm going to
match the DME as close as possible to the style and just pitch it.

[tschmidlin]

If you make your bug cultures at blood temperature (37C) or warmer, then you will get closer to the 30-45 minute estimate.  I think someone already posted something relative to these temperatures in their protocol above.  However, if you are simply making a starter in your cool basement, then you are right and you probably will be closer to 2 hours per replication. 

Do you have a source for these times?  Because according to the author of this paper (Optimization of growth conditions for intensive propagation growth development and lactic acid production of selected strains of Lactobacilli) they were unable to get under 2 hours with any of the strains tested.  The tests were done at 35C for most strains and 37C for L. delbruckii, where they saw the fastest growth.  I note though, that this paper is from a conference and not peer-reviewed, so if you have a better source of info please let me know.

[wildknight]

Lactobacillus is a facultative anaerobe, so it can switch between aerobic and anaerobic metabolism.  I think the main difference is that in this study, they used pressured tubes to prevent oxygen from entering and forced the Lactobacillus to grow anaerobically.  The growth rate of bugs during fermentation is significantly reduced, as ATP production from anaerobic respiration is up to 100 times lower than aerobic respiration.  In our examples, we are using stir plates aeration devices in our starters (usually), where oxygen is infused continuously, which allows for greater growth rates. 

My experience with Lacto is limited to homebrew use, but with in 24 hours I generally see a lot of growth in stir plate starters, but I do not have any cell counts to back it up.  In our lab, we work with all kinds of environmental bacteria, Pseudomonas, Bacillus, Mycobacterium, etc., and we have no problem growing up cultures with 1-2 million cells per ml overnight, just starting from a very small starting inoculum. 

[tschmidlin]

Good point about the O2, that should speed things up.  24 hours is a lot of time to double, you can hit 1M cells/ml starting with less than 250 cells/ml if it only doubles every 2 hours.  I haven't done any counts either, it would be good to test, or at least find a relevant reference.

[gandelf]

It's been three days since I pitched the 1 liter Lacto starter; it dropped the pH from 5.56 to 3.31. So, I did a boil
to terminate the lacto, added the hops and then the 1056. Hmm, what else can I do to this? Wring hands. Maybe
some organic cranberry, or some homegrown raspberries?