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 SpitballJedi
Ancient Astronaut
 
Registered: 10/13/12
Posts: 8,598
Loc: Nibiru 
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 Re: Endospores in Cerial Grains Discussion  [Re: blindingleaf]
#21605817 - 04/27/15 07:59 PM (8 years, 11 months ago) |
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Quote:
blindingleaf said:
Quote:
I think we don't see signs of bacteria on dry grains because there just isn't enough moisture for favorable conditions to germinate. They won't just germinate just because they are in the presents of moisture.
but, if they need to germinate, then isn't that either an endospore, or something like it? the grains we get are often stored for half a year before we receive them. if there is bacteria on there that does not form endospores for long term survival in less than ideal conditions, i'd be curious to learn what kind of bacteria that is
I didn't clarify in that quote, my bad, but I was talking about endospores not germinating.
As far as the other stuff growing in the water, I think of it like this: There's so much shit floating around that does not produce endospores. I think we are so inundated with live particulates, that there is just no way to keep your grains from being covered by the time it gets from the farm to your bucket. I think a lot of stuff can still survive in the slight amount of humidity in the air, even the stuff that doesn't produce endospores, but it flourishes in the presents of abundant moisture.
I'd be interested in the ratios of endospore producers vs non-producers that are growing in the soak water. Bodh has the tools to do it I think. You up for a challenge?
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blindingleaf
blue collar underworld
Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Endospores in Cerial Grains Discussion [Re: SpitballJedi]
#21605854 - 04/27/15 08:05 PM (8 years, 11 months ago) |
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yea, i'd be curious too!
already got the sugar water, and sugar water/peptone i mixed with my hands (no heat except from tap water) started
next, ill dump some grains into a few sterilized qt jars in front of hood.
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A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!
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bodhisatta  
Smurf real estate agent
Registered: 04/30/13
Posts: 61,891
Loc: Milky way
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Re: Endospores in Cerial Grains Discussion [Re: blindingleaf] 1
#21605878 - 04/27/15 08:10 PM (8 years, 11 months ago) |
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 just want to thank BL and you too SBJ for being able to discuss without butthurt. A lost art
Im glad you're doing this experiment BL
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blindingleaf
blue collar underworld
Registered: 07/19/13
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21605894 - 04/27/15 08:13 PM (8 years, 11 months ago) |
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yea, it will be cool to see!
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A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!
The whole is greater than the sum of its parts
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SpitballJedi
Ancient Astronaut
Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: Endospores in Cerial Grains Discussion [Re: SpitballJedi]
#21605906 - 04/27/15 08:14 PM (8 years, 11 months ago) |
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Quote:
SpitballJedi said:
I'd be interested in the ratios of endospore producers vs non-producers that are growing in the soak water. Bodh has the tools to do it I think. You up for a challenge?
Is this something you could do if you had the time?
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bodhisatta
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Registered: 04/30/13
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Re: Endospores in Cerial Grains Discussion [Re: SpitballJedi]
#21605988 - 04/27/15 08:29 PM (8 years, 11 months ago) |
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Quote:
SpitballJedi said:
Quote:
SpitballJedi said:
I'd be interested in the ratios of endospore producers vs non-producers that are growing in the soak water. Bodh has the tools to do it I think. You up for a challenge?
Is this something you could do if you had the time?
I could use plate count agar for total enumeration but it would be hard to get data you can translate into something meaningful. I'll ask around I still can get in touch with some people that might be able to give some pointers on things like that
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Pastywhyte
Say hello to my little friend
Registered: 09/15/12
Posts: 37,843
Loc: Canada
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21606027 - 04/27/15 08:39 PM (8 years, 11 months ago) |
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Can't read it now but I will be back to cheak out what I'm sure is a great thread.
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SpitballJedi
Ancient Astronaut
Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21606168 - 04/27/15 09:06 PM (8 years, 11 months ago) |
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Seems like it would be a lot of work trying to identify species and getting a count. It would be useful to know though. I think it would help shine some light on some of the debates related to endospores and soaking.
We mostly go off what other smart people say, what we know about general endospore reactions, and then fit that in to why we think we are supposed to do stuff.
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bodhisatta
Smurf real estate agent
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Re: Endospores in Cerial Grains Discussion [Re: SpitballJedi]
#21606181 - 04/27/15 09:08 PM (8 years, 11 months ago) |
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I think a good starting point is doing some grain jars sterilized for 30/45/60m (genrally less than recommended) using cereal grain with a "endospore soak" and without. but the grains are both going to be hydrated appropriately first. I do suspect the "endospore soak" grains contaminate before the no soak grains do.
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blindingleaf
blue collar underworld
Registered: 07/19/13
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21606193 - 04/27/15 09:12 PM (8 years, 11 months ago) |
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yea, i agree, thats the best starting point.
I'm down with shorter cycle too, at least first. say, 60 min, then done, for each pre style (soak/simmer, simmer only, dry grain + water), see the results, then increase to 90, and see what happens.
incubation at 80 u think? just to make things go faster?
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A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!
The whole is greater than the sum of its parts
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bodhisatta
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Re: Endospores in Cerial Grains Discussion [Re: blindingleaf]
#21606213 - 04/27/15 09:17 PM (8 years, 11 months ago) |
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I don't want to build an incubation chamber but they'll at least both be same temp. Its almost summer so hot soon
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SpitballJedi
Ancient Astronaut
Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21606244 - 04/27/15 09:22 PM (8 years, 11 months ago) |
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I may try to set up that experiments this weekend with the rye I have left. I've done soak, no-soak, boil, simmer, hot bath with rye and WBS and left all of them out on the counter for weeks after PC. They all did the same thing, dry without contamination.
I've already got my method for no-soak rye, so I know I can get it hydrated properly. Well, it's not a long soak for endospores at least. It's based on foomans WBS but with a longer time at a lower temperature.
I'll do it at 70-75F because that's my cult room temp.
I think the short time it takes to rinse simmer/boil, drain, and get the PC up to pressure is enough to hydrate endospores enough to get them soft enough to make them easy to destroy in the PC, especially for 120mins at 15psi. That's just theory based on personal experience.
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bodhisatta
Smurf real estate agent
Registered: 04/30/13
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Re: Endospores in Cerial Grains Discussion [Re: SpitballJedi]
#21606596 - 04/27/15 10:32 PM (8 years, 11 months ago) |
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My temps are 66-68 right now.
I usually do my rye throw in a pot of cool water rinse once bring to a simmer on the lowest setting takes about an hour then bring to a boil strain and don't make as dry as you normally would for long soak grain
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 Sarah Tonin
Gardener Gone Astray
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21607020 - 04/28/15 12:27 AM (8 years, 11 months ago) |
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 nerdy stuff to watch 
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blackdust
Registered: 02/28/09
Posts: 8,327
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21607855 - 04/28/15 07:01 AM (8 years, 11 months ago) |
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added for reference
 
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blackout
Registered: 07/16/00
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21609492 - 04/28/15 02:51 PM (8 years, 11 months ago) |
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Quote:
bodhisatta said:
my understanding of tyndallization was that yes they are short intervals of 12 hours between treatments but the overall time of all three treatments is greater than 48 hours to ensure that all endospores have germinated.
If you go 48hrs with only 3 heat treatments then at least one of them has got to have been over the 12hour stated time frame, after which was said would lead to too much bacterial growth...
If any side by side tests are to be done I would be interested in seeing the protocols involved, it drives me mad watching the likes of mythbusters and screaming at the tv how it was doomed from the start, or a completely unfair comparison. Fully hydrated grains was mentioned, I first thought the comparison was to be against grain with just water added before the boil. If not how is this hydration done? You can easily monitor hydration levels by weighing. However if you soak grains and then simmer to get to say 50% moisture content, then if you use unsoaked grains you would typically have to boil longer to get to the same moisture content -so it has undergone a longer heat treatment prior to sterilization. As I said before the recommendation of soaking is not solely about endospore germination, we must not jump to conclusions about the benefits which might skew test methods.
There is lots of advice on soaking grains for human consumption these days, many recommend changing water to stop any overgrowth of bacteria. I have soaked grains for over a week with regular enough changing of water. I would often drain and pour on boiling water.
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bodhisatta
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Re: Endospores in Cerial Grains Discussion [Re: blackout] 1
#21609717 - 04/28/15 03:53 PM (8 years, 11 months ago) |
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well what would you do and how would you set it up blackout?
I get fully hydrated grain in under 2 hours when I do a no soak but the simmer and boil are a little longer.
We do know from empirically gathered evidence that
soak or no soak you can cook grains effectively thus the PC kills endospores just fine.
all I'm trying to figure out is
if no soak grains have less or more endospores than soak grain right before you are about to sterilize.
not about tyndall and his method. but if you read tyndall's method it does lend evidence to the side of the argument that you very well could have more endospores in the soak grain since soaking grain for the explicit purpose to germinate endospores doesn't coincide with tyndall's suggestions to ensure that you don't cause that problem. Tyndall really has nothing to do with this but since you brought it up eariler I just thought it was funny that if you read his real methods and not other people's interpretations it does basically say that endospores can take up to 2 days to germinate and if you soak longer than 12 hours you end up creating new ones. is this true? I don't know but if we assume it's true tyndall is more on the side of the argument that soaking for the explicit purpose of germinating endospores is moot, and if the soak is doing anything beneficial it's not beneficial for that reason("endospore softening").
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blackdust
Registered: 02/28/09
Posts: 8,327
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Re: Endospores in Cerial Grains Discussion [Re: bodhisatta]
#21609843 - 04/28/15 04:31 PM (8 years, 11 months ago) |
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Log in to view attachment
I wanted to add some research I found. I started to read the review but havn't finished it yet.
Here it is in pdf format. I found it through my schools article search
Source: Nature Reviews Microbiology
Date: January 1, 2013
The Bacillus subtilis endospore:assembly and functions of the multilayered coat
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blindingleaf
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Re: Endospores in Cerial Grains Discussion [Re: blackdust]
#21610254 - 04/28/15 06:23 PM (8 years, 11 months ago) |
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the main thing i don't understand is
if u think the soak is not enough time for endospores to germinate (i disagree), then how are there more endospores at the end of the soak if they are being created in the process of bacterial reproduction?
are there any references to how long it take to germinate (i would guess based on temp)?
in the black dust article above it says "Spores are capable of quickly germinating and resuming vegetative growth in response to nutrients"
which i would agree with based on soaking grains.
i would guess not all of them germinate as fast as each other, but by hour 4-6 with a hot water soak, i'd guess many would be germinated.
then the question is, how long does it take the process of replication to take place?
in the other thread in MC (locked) i included a quote/site that said endospore formation takes anywhere from 8-10 hours.
so 8-10 hours, plus 4-6 hours (initial germination) leaves us at btw 12-16 hours, basically as long as a standard overnight soak.
i still don't think that there are more endospores at the end of a soak. and if there are, I'm not sure why i get less bacteria now then when i first started.
i also still think the grain + water should be included in the comparison, because the grain is going to be submerged (standard ratio in this case for 1 qt is 200g dry rye:220ml water).
and since u think the autoclave PC is capable of destroying endospores, then i would think that would count? the bacillus strips u mentioned in the OP, i wouldn't think those are wet, are they?
I'm curious why u choose 2 hours for a PC time instead of less? what happened when u did one hour, or 1.5 hours? surely something was surviving.
i think we each may have different experiences with bacteria.
i had a lot of bacteria at first when i started.
i think there is variation in endospore counts from grain to grain, and source to source.
when someone hasn't had many bacteria problems, i think the endospore thing doesn't seem as big a deal as it could be.
i'd like to hear what pasty has to say as well, because i know we both shared many a bacteria problem (we referred to them as endospore problems though, i guess you would not agree)
but to think a soak does not germinate endospores just sounds wild to me, i dunno.
i'd really like to see something that talks about long endospore germination times that you are referring to.
the water plus sugar was still the same, no smell or anything (45g sugar:200ml water)
same with the peptone one
both were started with hot tap.
--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!
The whole is greater than the sum of its parts
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drake89
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Re: Endospores in Cerial Grains Discussion [Re: blindingleaf]
#21610544 - 04/28/15 07:18 PM (8 years, 11 months ago) |
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I think there's a big difference between feed vs seed vs human food grade grains. The feed grade grains are dirty. Seed grade is a bit better but human food grade is best and cleanest. I think you may have some bacteria all the time but at some point they can take over. Just my take on it.
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