Loss of second generation yeast?

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ObsidianJester

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Well, I'm not sure really how to do a search for something similar so I'll describe what I observed.

Few weeks ago I brewed up a cream ale type of brew, heavily hopped and used Wyeast 1056. Got some great yeast to harvest out of it with very little trub to speak of.

So I saved this and used it for the same grain bill the second go around, only used different hops and a bit more water. Everything else was the same, the temps, boil time, irish moss addition, using a yeast starter. This time around I suspect due to the bigger water amount I had yeast krausening out of the air lock in the bucket. No biggie, just clean it up and replace the air lock. The fermentation took longer which I would attribute to there being about half a gallon or so more wort than the previous time. Let it sit for the two weeks, and got on with bottling. However when I went to harvest the yeast by a normal addition of some cooled boiled water I did not see much yeast. Quite a lot of hot/cold break material, trub but not a lot of the visible yeast.
The amount I pulled was three quart mason jars worth of material. The first was nearly all liquid with perhaps a few scant millimeters worth of settled yeast. The second jar had much of this trub material in it, about a centimeter at best of the typical yeast layer and half full of the liquid.
The third jar had all trub, a very thin layer of the yeast and then liquid.

I understand that there are studies and experiments that state active yeast is in all three layers, though adding the trub is more dead yeast, and you don't want to have too much of that in your next brew. Decanting the liquid layer is personal preference but to keep off tastes out of it it is generally decanted or the large bulk of it is.

I also understand that I can grow up a starter with what I can harvest out of the two viable looking jars. I plan to do so with this first (Second?) generation on Friday the 21st with anticipation of brewing a rye style ale on Saturday afternoon.

I'm just curious as to what happened to change the dynamic up. What could have happened to the yeast? The ale seems to be bottle conditioning normally. Even if something had changed in the brewing to create more of the possible hot break material I would think the yeast would still be showing in abundance. Anyone experience this before? The ABV is not different either if that helps.

What happened?

What the heck happened?
 
I am also curious what people have to say because I also just washed a wyeast 1056 and wondered how many times I can rewash each pint of the washed yeast I harvested and if it'll still work the same
 

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