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OfflineL0ki
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Registered: 06/22/06
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Last seen: 16 years, 9 months
PPP - Partial Pressure Pf
    #5927534 - 08/03/06 12:33 AM (17 years, 7 months ago)

Hello All,

Been having probs with endos. I usually grind up organic brown rice, ( in a coffee grinder), with really good success, but lately the batches have been going sour due to endospores (MOST likely in the rice, but can it be in the vermic? the air? or bleached down poly on the walls? how hard is it to kill existing enviromental endospores in a room that housed a tank covered by a rubber seal and glass that held the vermic bedded yet sourly contaminated pf garden?? is this a long parentheses? :eek:).
If i were to use the pf method, say in a 36 250ml jar capacity pressure cooker, what should be used as cooking times using the partial pressure method? I know using this method involves an equation, but I was thinking at about 2 cooks @ 45 minutes (15 psi) with enough time between cooks to add more water to canner so it doesnt dry out. close-to-boiling water at that so jars dont crack. anyways, thats just a guess to kill the endospores, cause for some reason where I am, I cant seem to soak anything over night and kill it off for the life of me. 3 hours (at 15) couldnt do 1 litre of wbs in two jars, soaked over night or not ( i did it twice), so im ready to try partial sterilization, but i really need some clarifications on cook times vs. volume vs. pf method & agar (i was getting away with 20min/20min @ 15 but no longer and im scared of caramelization).

Are the restart times/temps critical to the probability of risk as well, or can i just get it to a handleable temp and start cooking again? 

Also, has anyone know if a hepa filter can become a housing for contamination even if teh cartridge is still white, but ran for a while?

sorry so asking, been at work tooooooo long with sour myc on my mind. :mushroom2:
this is also just waaay too much to query on the search posts, and after fifteen different keyword combinations and hours of reading, ive had enough, and gave into the posting this question, with all its crazy details.

but any help would be appreciated. :crazy:

L0ki

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OfflineRogerRabbitM
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Re: PPP - Partial Pressure Pf [Re: L0ki]
    #5927578 - 08/03/06 12:54 AM (17 years, 7 months ago)

Grains harbor endospores, but rice rarely does, which is why the pf tek can usually be successful without a PC. I doubt they're your problem. An hour at 15 psi should be plenty if there's ample room between the jars for the steam to penetrate. Be sure to let the PC vent steam for several minutes before you put the weight on or close the valve. It's necessary to heat the interior, including the glass surface of the jars, and vent out all the air before building pressure.

That said, there is bacteria in the air everywhere. If your verm barrier is damp, the bacteria can colonize right through it to the rice flour below. Perhaps your rice is too wet, or you're not getting the jars properly covered before sterilization? Remember, when the steam hits the cooler lid of the PC, it turns back to water and rains down on the jars. This is the reason for the foil covers.

Your PC should never run dry even during a three hour cycle. Be sure to reduce heat as soon as it reaches pressure to the minimum setting that will maintain the 15 psi. Very little rattling of the weight should be allowed. If the weight rattles, turn down the heat slightly. If pressure drops below the target 14.5 psi, increase heat slightly.

It actually sounds like either your verm is getting wet/damp, or perhaps your syringes are getting contaminated. Remember also, one of the biggest sources of bacteria is your own breath, so wear a surgical, not dust mask. A close second is your skin and especially under your fingernails, so wear gloves that have been washed with alcohol.

If you're 'incubating' your jars, have plenty of ventilation. If they're in a closed up container, the humidity can cause your filter material to get wet, rendering it useless. I recommend incubating on an open shelf at room temperature for the reduction in these problems. The extra two or three days it takes to colonize a jar is worth the reduced risk of contamination. Good luck in finding and fixing the probelem.
RR


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InvisibleOmnicracker
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Re: PPP - Partial Pressure Pf [Re: RogerRabbit]
    #5927597 - 08/03/06 01:01 AM (17 years, 7 months ago)

RR, when you say "surgical, not dust mask" is there a difference in the material they are made of, or its is just a matter of how they fit/froce air throught the filter?
http://www.uline.com/Browse_Listing_952.asp?pricode=wj896

i use nice fitting dust masks, though it looks like surgical masks might be more comfortable...


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OfflineL0ki
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Re: PPP - Partial Pressure Pf [Re: Omnicracker]
    #5927702 - 08/03/06 02:38 AM (17 years, 7 months ago)

to quote RogerRabbit - "If your verm barrier is damp, the bacteria can colonize right through it to the rice flour below. Perhaps your rice is too wet, or you're not getting the jars properly covered before sterilization? Remember, when the steam hits the cooler lid of the PC, it turns back to water and rains down on the jars. This is the reason for the foil covers."

Hear ya there. I always foil, and honestly, that vermic is pretty dry up there when i poke in to innoc. I found the best weapon is a meat injector. 2 bucks, never clogs, GFS. But even dry, i've found some have gone sour lately. Hence the rice contam possibility.

But, if it is, i need to be absolutely sure all traces of it are gone.

This doesnt explain the stuff dissection cant answer (like agar, STRAW, wbs (mabey the peanuts?)), almost everything in that room has gone the same exact sour, and im scared the source was the rice itself. what would you recommend the cleaning procedures for this type of endospore to reduce the risk at a reasonable level? should i be afraid of nearby carpets, that kind of thing? or will a simple bleachrag do the trick? I've also found spraying lysol alot doesnt help either for this (in fact, it smells WORSE after the lysol... sickly, then later on even more sour, but cutting off lysol dont help it after bleachin none either... konundrum? fuck it, im buying some OUST tommorow). i am serious about all the poly though, and i bleach teh shit out of it 1 a month for good measure. but 3 months now and STILL not gone. man i had it raining in there with bleach water and then bleach-wiped all the equip back in. That HEPA runs all the time, usually on silent or half blast. The filter is a concern too but its HEPA, so again, should it be replaced or is it waistin $$$ (20 bucks i kind of need to know if i should spend). its the oulying area i worry that is tracking in..., but theres nothing alive there of noting, or anything smelly, only at the vicinity of the garden.

Also, from RogerRabbit "Your PC should never run dry even during a three hour cycle. Be sure to reduce heat as soon as it reaches pressure to the minimum setting that will maintain the 15 psi. Very little rattling of the weight should be allowed. If the weight rattles, turn down the heat slightly. If pressure drops below the target 14.5 psi, increase heat slightly."

good point here too, sometimes i've hit 16-17 psi (by sheeeer ignorance), but no issues there... these last few times i was a hawk so it sat really good on 15 - 15.5 thats close enough i should think, but is the added heat a good thing in terms of sterilizing? does the material matter as long as the material wasnt pooling? those wbs seeds had polyfill, with coffee filter cover, and sat shelved with aluminum covering up to the lid. Mason style, 1/2 hole. 3 hours still had water left so that was good too... thinking of tyvek next, but again, scared its the inside enviroment (hence partial pressures...). in tank incubation doesnt help either. neither upside-down mason lid method. i've made it REALLY dry (NONE of the seeds stuck, usually theres one or two) and tried the pasty.. 4-5 cooks in all in just experiments with wbs & PF& agar, in the same room, 1 tank per type, for incubating. 24 dark, at ~80F. all contamed sour and with this type of contam, its hard to see where it starts, cause it reeks before it shows, like all endospores (damn). then youre adjcent cultures contam and you never know which one it is without risking destroying the now dying crop anyways.... sigh. which is why im posting at 4:00 am.

more quoting... - " Remember also, one of the biggest sources of bacteria is your own breath, so wear a surgical, not dust mask. A close second is your skin and especially under your fingernails, so wear gloves that have been washed with alcohol."

the mask i'll try, strange though, never had that prob with the breath before (i always shower and listerine first before innoc), but why not? paranoia pays off as i like to say, and HEPA's the way to go, dustmasks are shit for the resolution for bacteria. whats the expected diameter anyway? less than 20 nm?(again, ive never had to before... sigh).
the gloves, well i usually go with fresh-from-washer-&-dryer long sleeve shirt, "sterile" marked medical gloves and then hose them down while wearing with lysol. get the sleeves too. Repeat after 15-20 innocs. Tank with a sheet of glass over 90% of it. stick hands through smallest hole possible after lysoling (again fuck it OUST tommorow baby) the snots out of the box and a good 30 seconds with the lid completely closed. worked for years for pf, but i've never been able to be successful with anything else. now, its not cuttin the mustard even for pf, so im freaked out. theres is a possibility to rig a flow hood, but that makes no sense if i cant clean the surroundings correctly. how long can bacteria live on dry, soft and hard surfaces? like brick and carpet?? is it possible? or do just gotta hit it with alcohol & oust instead this time?

L0ki

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Invisiblefastfred
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Re: PPP - Partial Pressure Pf [Re: Omnicracker]
    #5928065 - 08/03/06 09:14 AM (17 years, 7 months ago)

Quote:

Omnicracker said:
RR, when you say "surgical, not dust mask" is there a difference in the material they are made of, or its is just a matter of how they fit/froce air throught the filter?

i use nice fitting dust masks, though it looks like surgical masks might be more comfortable...




There seems to be some misconception over the difference between surgical and dust masks. As odd as it may sound dust masks are usually better than surgical masks.

Surgical masks are designed to protect the wearer from splashes of bodily fluids entering the nose and mouth, and spit and mucus from contaminating the patient. They don't seal well to the face and are not terribly effective at arresting particles.

Dust masks are designed to arrest particles and make a better seal to the face than surgical masks.

Here are some quotes from some websites...

"Surgical masks do not have either adequate filtering or fitting attributes to provide respiratory protection for the wearer. They are designed to help prevent contamination of the work environment or sterile field from large particles generated by the wearer (e.g. spit, mucous). Surgical masks may also be used to help reduce the risk of splashes or sprays of blood, body fluids, secretions and excretions from reaching the wearer's mouth and nose."

"Surgical masks are not designed to seal against the face and are not fit tested since the main goal is to help capture large particles expelled by the wearer and to help reduce the wearer's exposure to splashes. During inhalation, much of the air passes through gaps between the face and the surgical mask."

"The Food and Drug Administration (FDA) does not test surgical masks. Rather, the manufacturer provides data and proposed claims to FDA for review. The FDA reviews manufacturer's test data and "clears" the mask for sale."

"The filter media of a surgical mask with a very high PFE (>95%) may be less than 70% efficient when tested with the NIOSH N95 test method. Additionally, because surgical masks do not seal against the face when worn much of the inhaled air passes through gaps between the mask and the face rather than through the filter media."

"Surgical masks are not designed to pass a fit test. Flat surgical masks fit loosely over the face leaving large gaps between the mask and the wearer. It is unlikely that most of the air will pass through the mask material. The air (and any airborne particles) will go through the gaps."

"In the United States, surgical masks are cleared for sale by the Food and Drug Administration (FDA). The FDA reviews data submitted by the manufacturer but does not test the surgical mask."


For dust masks and respirators...

"Look for a NIOSH rating of at least 95 ... this will be normally listed as N95. The 95 represents a 95% efficiency on a rigorous standardized test. N 99 is a higher rating than N95, and N100 represents the highest rating."

"While there are comfort and other differences between brands, any filter or mask with a NIOSH certification will do the job adequately."


What you want is a dust mask that has been NIOSH certified as a respirator. A N-95 certified dustmask/respirator is almost twice as effective as a surgical mask for arresting particles that go through it, and it fits well to the face whereas a good portion of the airflow will go around a surgical mask.

Here is a good one that I've used. A lot of garden and hardware stores will carry them.

3M™ Lawn and Garden Respirator 8659ES N95

The N-95 certification makes them cost a buck or two each.

In one test on filtration efficiency a bandanna folded twice filtered 6%, folded 4 times filtered 10%. A surgical mask filtered 62%, and a N-95 mask filtered 98%.

If you buy a cheap hardware store mask that costs around 50 cents each they will range from about 13% to 95% depending on quality, but you'll never know what you're getting unless they're certified. Sometimes you can get a good deal on masks weren't certified but later got certified. They have to sell those at regular price since certification is not retroactive once it leaves the factory.

The main purpose of wearing a mask is to slow and diffuse your breath so that you're not actually breathing on your work, not to actually filter it. So any mask will do a decent job of assisting your sterile technique. If you actually want to rely on the mask to filter your breath then you need to get a N95, N99, or N100 mask. A surgical mask is no good compared to them.


I hope this helps and clears up any disinformation and/or misinformation that's out there.


-FF


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It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy

The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed

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I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid

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OfflineL0ki
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Re: PPP - Partial Pressure Pf [Re: fastfred]
    #5932257 - 08/04/06 05:56 PM (17 years, 7 months ago)

thanx for the reference on the masks. i wasn't aware of the whole N-guaging, but after a little reasearch it looks worth the shot. can anyone say anyting about kiling endospores and where they can/cannot live or should i leave that to another topic thread?

thnx
L0ki

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OfflineRogerRabbitM
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Re: PPP - Partial Pressure Pf [Re: L0ki]
    #5932389 - 08/04/06 06:52 PM (17 years, 7 months ago)

Wait.

I said you want to use surgical masks, not dust masks. That is correct. Surgical masks are designed to protect a patient from the bacteria being exhaled by the doctor during surgery, which is exactly what we need a mask for. The surgical masks I get from a local drug store are clearly labeled "stops 99% of the wearers exhaled bacteria"

Our mycology projects are not going to attack us with large partical size dust, such as floor sweepers or or someone running a concrete saw are going to be exposed to, but the bacteria we exhale with every breath can easily contaminate our needles, scalpels, inoculating loops and grain jars, as well as the whole general area we are working in. Bacteria are single celled creatures and extremely small. It takes a microscope to see them. The bacteria in your breath will sail right through a dust mask like a gnat through a fishing net. The micron size of a dust mask is simply too large to be effective against bacteria. That's why they're not used in hospital surgery rooms.

Endospores are a survival tactic used by bacteria. They are resplendant in grains, such as rye, wbs and corn. If you'll soak your grains for a few hours to hydrate the endospores(as well as the grain kernels), they'll be easily killed in the PC, even if they haven't germinated yet. Sometimes they survive the pressure cooker if that procedure is skipped. Good luck.
RR

edited because I hate typos....lol


--------------------
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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

Edited by RogerRabbit (08/04/06 07:15 PM)

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Invisiblecappa
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Re: PPP - Partial Pressure Pf [Re: fastfred]
    #5932391 - 08/04/06 06:53 PM (17 years, 7 months ago)

Going to do my own research on this subject.

Thank you both for potentially exposing each side of the issue.


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Edited by cappa (08/05/06 09:13 PM)

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OfflineL0ki
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Re: PPP - Partial Pressure Pf [Re: cappa]
    #5941603 - 08/07/06 06:01 PM (17 years, 7 months ago)

hey, sorry so long, work sucks...

well, looks like a did a few tests, and im taking pics and u/ling them sortly. if the case is definately bacteria (which it seems, but pics tell a 1000 tales), is oust, lysol, bleach and alcohol good enough for the cleaning process? normally i just use lysol and bleach, but its funny as i looked at the ingredients of the aresols today

oust is ethanol & triethylene glycol
lysol neutra-air is just triethylene glycol
lysol is ethanol, Alkyl (40% C12, 50% C14, 10% C16) and finally Dimethyl Benzyl Ammonium Saccharinate 0.106%

now, i understand the ethanol application, cuase thats basically alcohol.

the alkyl is an agent made specifiacally for contams killing, but mainly for surfaces, as they have no way of stayin in the air themselves, and the triethylene glycol seems to be a suspending agent / another form of alcohol (must be some form of sanitizing agent as well, as its the only active ingredient in lysol neutra-air). the rest, not too sure, but light shed on that subject ( or any corrections to my understanding) would be appreciated. This bacterial contamination bullcrap has haunted me long enough. been dying to do a brf -> straw ball but everything i've tried goes sour, so even after bleaching, i get the same results (argh). so its, these three products, and of course the surgical masks (not the dust). Besides this, and wiping everything down with bleach & alcohol (isopropyl, 99% or 70%).

Also, pics coming shortly, need to show you if im doing this right... wish i could change the topic of this posting to "damn im a noob"... sigh


L0ki

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