Quick question about sterilisation

[Hillbilly32]

Is using a pressure cooker the only way of killing the bacteria or are there other means, as I do not currently own one.

Also, is spraying dis-infectant into jars a bad idea? I am thinking that it would kill the mycelium, would that be correct?

Thanks

[worowa]

Many ways-e.g. peroxide...try the shroomery, lots of ideas there.

[Hillbilly32]

Already had a quick look, and most teks use pressure cookers. I have dis-infectant, could I use that?

[ENtiTY]

Every thing needs to be sterilised. Not just the jars but the grain or whatever your using to grow the myc on needs to have any nasties that may be in it killed as well. Only way to do this is with heat and pressure, i.e. pressure cooker or auto-clave. If your on a budget try looking at garage sales and markets etc. Amazing what you can find there

Those old thick walled aluminium PC's are better than the new stainless steel PC's anyway. The older ones run at close to 15psi which is what you want, where as the new SS ones only run at about 7psi or so unless you want to pay a lot more. You want one with a capacity of at least around 10L.

Good luck

[apothecary]

Your question is "can I take the lazy way out".

Newbies always make the mistake of thinking they are the first person to think of a "shortcut", if it was a functional and more efficient technique then all those writeups on the shroomery wouldn't bother mentioning pressure cookers?

The answer for all sterility shortcut questions like this is no. You need to follow correct sterile procedure to get anywhere.

[Ace]

I agree Sina, but a lot of the time it involves cutting down on expenses, not neccessarily blatant laziness.

HB, like Harry mentioned, look in market stalls, second hand shops, vinnies/salvos, garage sales etc. They are around and can be found cheap. Keep an eye out and you'll be PCing all your stuff in no time. If you can afford it, then you might want to be lazy and get a brand-spanker, but like it has been mentioned the older ones are far better as far as pressure goes.

That said, I'm sure there have been a few write ups where a regular pot of boiling water was used (for a very long period of boiling) instead of a PC/autoclave, but results were rather poor (as you would expect). If you are too lazy/poor to get a PC (understandable), then perhaps try this as a last resort.

Remember with patience comes fruition.

[Hillbilly32]

In this thread:

http://www.shaman-australis.com/forum/inde...showtopic=14230

Hyphalknot just puts boiling water into an Eski with his jars in it to perform the sterilization. I was thinking, what is wrong with just plunking them in the oven on low for half an hour or so? It would definetly kill the bacteria as the temperature will be up around 160° C?

[Ace]

I havent bothered clicking the link (sorry, a bad mix of laziness and busy-ness ), but I have a strong feeling that Hyphal wasnt sterilising, but actually pasturising (slightly different terms/processes) in the eski tek. Dont quote me, but going from memory and the sound of the tek, that'd be what he's referring to.

BTW, I'm almost positive that there are some bacterias that could survive 160C. Couldnt name any, just something I am pretty sure of

[Hillbilly32]

Yes sorry, you are right, it was pasturising. What is the difference? I know that doing it this way it does not kill 'all' the bacteria, though he also notes that its only good bacteria left over which kill off any other bad bacteria. Isn't that better than killing 'all' bacteria?

[Ace]

What is the difference?

To be honest, I never got that far into things to do the research on pasturising. I think there is a pinned thread (possibly the one you linked) that explains pasturising, bulk substrates and casing. If you cant find it, maybe just have a search for 'pasturising'. Would you mind posting the answer, if you find it?

[ENtiTY]

Yes sorry, you are right, it was pasturising. What is the difference? I know that doing it this way it does not kill 'all' the bacteria, though he also notes that its only good bacteria left over which kill off any other bad bacteria. Isn't that better than killing 'all' bacteria?

It is better, that is why pasturisation is used at that point. You see the pasturised material is going to be used to spawn the colonised grain to. First you have to get to the point where you have colonised grain. You will likely be starting from spores off a print, to grow a healthy uncontaminated grain spawn you have to kill everything before you innoculate it to give yourself the best possible chance of sucess. A healthy, established, colonised grain can out compete contamination where germinating spores cannot.

My suggestion is, before you attempt anything read, read, read and read some more on the subject. You need to clearly understand the processes of growing from start to finish and understand why each process needs to exist. Once you have an understanding based upon actual experience then you can tweak, dabble and experiment with improving growing techniques. You need to have a sound knowledge base before you attempt any refinement, otherwise you are doomed to much frustration and failure.

When you think you have a grasp of whats happening, pick a tek thats tried and true and start from there. I'd suggest the PF tek as thats the easiest to start with but thats usually used for growing cubes which are illegal so that advice is prolly no use to you

Time to become a bookworm Mr Hillbilly

[worowa]

I've had 2 successfull experiments using no PC.

The first was while cooking dinner-I had left over brown rice, not PC'd, just cooked for dinner, and I had the big butt of some "shimeji" mushrooms (probably Hypsizygus ulmarius). Well, in my not very clean kitchen, I half filled a plastic food container with the rice, then shoved in the butt (half the size of my palm), put a lid on, and off she went-no contams visible, and we had small shrooms straight off the rice sometime later. The culture lived in my car while I camped-no special care.

The second was with shiitake on hessian, the hessian was soaked in hot water, drained, soaked in peroxidated water, drained again, then chucked into a pot of boiling water for a minute and left to cool. I had been practicing cloning on agar, using the peroxide method, and put a chunk of myc on the hessian, and she took off. Hasn't fruited yet.

I use a PC sometimes, just got a BIG one , but I really don't think it's essential, just a more reliable option. As for disinfectants, there's so many different sorts, some nasty, some possibly cheap and useful...what have you got to lose experimenting? A couple of bucks for a bit of education. Mycotopia is also worth a look.

[Hyphal]

It is better, that is why pasturisation is used at that point. You see the pasturised material is going to be used to spawn the colonised grain to. First you have to get to the point where you have colonised grain. You will likely be starting from spores off a print, to grow a healthy uncontaminated grain spawn you have to kill everything before you innoculate it to give yourself the best possible chance of sucess. A healthy, established, colonised grain can out compete contamination where germinating spores cannot.

My suggestion is, before you attempt anything read, read, read and read some more on the subject. You need to clearly understand the processes of growing from start to finish and understand why each process needs to exist. Once you have an understanding based upon actual experience then you can tweak, dabble and experiment with improving growing techniques. You need to have a sound knowledge base before you attempt any refinement, otherwise you are doomed to much frustration and failure.

When you think you have a grasp of whats happening, pick a tek thats tried and true and start from there. I'd suggest the PF tek as thats the easiest to start with but thats usually used for growing cubes which are illegal so that advice is prolly no use to you

Time to become a bookworm Mr Hillbilly

You got it in one there Harry - to germinate spores and get them to completely colonise a substrate - you can't have any competition from other spores or bacteria. But, once you have fully colonised jars, you can spawn them to a pasturised substrate and succeed as your introduced culture will then be the dominant fungus and will therefore flourish.

You can however steam sterilise without the need of a pressure cooker, as long as you are only using the pf tek. NB - Grains CANNOT be sterilised this way.

Simply follow the pf tek for simple minds and instead of pressure cooking your jars - steam sterilise them in a pot with a tight fitting lid for 1.5-2 hours.

Method:

Clean you jars well with hot soapy water and a clean dish brush, then rinse well with super hot water and drain upside down on a clean tea towel before loading in your mix and adding the foil lids.

Once the jars are in the pot, just pour cold water in until its half way up the jars before bringing the lot to the boil, then drop the temp down so you get a nice, rolling simmer.

Wack on the lid and make sure its nice and tight - if need be put a weight on the top (eg. a rock or a bowl of water if it will balance) and wait 2 hours.

Check on your jars an hour in to the process to make sure there is still enough water in there, and top it up if necessary. (At this point, if you want to, you can suck up a syringe full of the

boiling water, wrap it in foil and put it in with your jars to sterilise it for later making a spore syringe).

After two hours, turn off the stove and allow the lot to cool overnight. Don't lift the lid until they are cool because as they cool they will suck in a bit of air. Inside the pot is a pretty clean environment after the boiling and opening it while they are cooling is just another opportunity for dirty air to come in, although it probably won't actually make much difference, you may as well not.

Innoculate the next day when cool and incubate!

You should achieve 100% success with this method if you follow sterile printing and syringe making techniques.

Lastly, pf style jars can be used successfully to grow other things than cubensis, eg. 'oyster mushrooms'.

Edited January 29, 2008 by Hyphalknot

[Hillbilly32]

Wow, thanks a lot guys, I really wasn't expecting you to go out of your way and into so much detail for a newbie like me

I actually ended up buying some soft plastic containers to hold the mixture in to sterilise, however, I now know that that was a mistake. I've got a bunch of really really strong glass jars that have a volume of about 750ml, but the neck is narrower than the body, so 'cakes' are out of the question unless I buy some new jars.

I have seen them grown in 2 different ways. In cakes, and in a pan. Now, I reckon a pan would be a lot easier to maintain and take care of, but I'm sure there are some disadvantages to it.

Edited January 29, 2008 by Hillbilly32

[Hyphal]

Follow that link mate, you dont need jars - any drinking glasses will work absolutely fine. Also, you can spoon colonised substrate out of your jars and case it if the necks are smaller than the bodies, but 750ml jars would take forever to colonise pf tek style so go for short, fat 200-250ml drinking glasses.

Read the link fully and you will have a heaps better understanding of what you need to do.

[Hillbilly32]

Just sterilised 3 jars for 140 minutes and injected the "oyster mushroom" spores *wink wink* into 2 of the jars while they were still warm, and the other I am waiting for it to cool down and then will do the same.

A couple of problems though. I only have a 1ml syringe without a needle, so I used Kirof Vodka as a dis-infectant (37% alcohol) which I don't think was strong enough, but everything else was relatively tidy and clean, and when it came to injecting the spores, I used a sowing needle which was also dunked in the alcohol and flame sterilized with a small blow torch to create the first puncture hole.

Now, because of this, I have a few quick questions:

1) Would some of alcohol that got left behind in the syringe have made a difference to the spores or the substrate?

2) Did it matter whether I let the jars cool down after sterilization or not? ie, does the heat destroy the spores?

3) Is methylated spirits ideal for cleaning? It is 95% alcohol so I don't see why not, yet no-one seems to use it?

Thanks

Edited January 31, 2008 by Hillbilly32

[Teljkon]

sdfsd

Edited December 19, 2021 by Teljkon

[ENtiTY]

Just sterilised 3 jars for 140 minutes and injected the "oyster mushroom" spores *wink wink* into 2 of the jars while they were still warm, and the other I am waiting for it to cool down and then will do the same.

A couple of problems though. I only have a 1ml syringe without a needle, so I used Kirof Vodka as a dis-infectant (37% alcohol) which I don't think was strong enough, but everything else was relatively tidy and clean, and when it came to injecting the spores, I used a sowing needle which was also dunked in the alcohol and flame sterilized with a small blow torch to create the first puncture hole.

Now, because of this, I have a few quick questions:

1) Would some of alcohol that got left behind in the syringe have made a difference to the spores or the substrate?

2) Did it matter whether I let the jars cool down after sterilization or not? ie, does the heat destroy the spores?

3) Is methylated spirits ideal for cleaning? It is 95% alcohol so I don't see why not, yet no-one seems to use it?

Thanks

For sterilization of the syringe you were probably better off using 3% hydrogen peroxide which you can get from woolise or pharmacy. Fill your syringe with it then leave it for 10 minues or so to do its thing, squirt it out then wrap the syringe in Al foil and drop it in boiling water for a few minutes. The heat will degrade whats left of the peroxide into harmless water so your spores will be safe. You should then have a reasonably sterile syringe. Never tried it as I use a PC for all my sterilization but if I didn't that would prolly be how I would go about it.

The alchohol may or may not effect the spores, depends on what concentration ended up in with the spores and for how long they were exposed. As a guess I'd say you may have slightly reduced you spore germination count.

Now syringes, go to a horse supply place or a pet shop that supplies for horses and ask for some 5ml and 10ml syringes. Then ask for some 1 1/2 inch 18gauge sharp tips to go with them. Get as many as you need, at least 1/2 a dozen of each.

Your better off leaving the jars to completely cool to room temp. The core of the cakes will be warmer than the outside so you have to keep this in mind. Heat will effect spores but more of a worry is the possible contam introduced along with the spores may get a head start with elevated cake temps at innoculation.

Some do use methylated spirits and it works fine. Works better if I remember correctly at about 70% methylated spirits and 30% water. The added water improves the ability of cells to absorb the alchohol or something like that. I think isopropanol is better at killing nasties but its more expensive and less common. Most of the tek's you will see are written by people in countries other than oz and iso is probably more common over there. Iso isn't really that hard to find if you want to go looking

[Guest Øskorei]

Your question is "can I take the lazy way out".

LOL, I thought the very same thing.

Newbies always make the mistake of thinking they are the first person to think of a "shortcut", if it was a functional and more efficient technique then all those writeups on the shroomery wouldn't bother mentioning pressure cookers?

The answer for all sterility shortcut questions like this is no. You need to follow correct sterile procedure to get anywhere.

Mate, a FOAF grabbed an old PC a couple of years back off E-Bay for about fifteen bucks (if at first you do a search and see nothing, wait a few weeks and try again). Take Apo's advice and run with the tried & tested success stories to home cultivation, you will find that it's not much more work, and you shall yield the rewards.

There ARE some teks that mention a simple saucepan but the success rate will be much lower especially if you cut cormers on the other considerations. Go with the PC mate, and all the best for golden days of happy fungulation Let us know how you go, eh ?

[Hyphal]

Well the first few times I sterilised pf jars was done with steam sterilisation and it worked fine, 100% success rate. I think the key is using smaller jars, therefore heating up more quickly, and the longer boil time of 2 hours seems to get the job done.

Those old style 'vegemite' jars worked a treat, but any small jar or glass about 200ml in size should be fine - at least to get you going until you save up for a good quality PC or find one cheap on ebay or in a second hand store.

To sterilise a syringe without a PC is easy and effective - get a pot of water boiling for ten minutes, then suck up and expell some of this boiling water in and out of a syringe several times (sucking up and squirting out into a sink - NOT back into the pot!), then on the last pull leaving the syringe body full of the hot water, wrapping it in foil and dropping it into the boiling water in the pot with the lid on for 45 mins works fine.

[Vertmorpheus]

Interesting thread... was wondering if anyone had any experience using the microwave? Certainly makes some nice clean prop mix, with very evenly dispersed water content, good penetrative ability and its hardly a single-use purchase (spousal harmony is priceless haha).

Also, has anyone fiddled with their own PC technologies? Retrofitted stock pots with gasket goo valves n taps? Put a 15 pound weight on a well fitting lid? Some mutant kind of double chamber bong theory? band-clamped surgical rubber sheeting over a soup pot?

VM

[Hillbilly32]

Not really what you were after Vertmorpheus, but I am sterilizing as we speak and I have just found that some string coiled around the lid and handles tightly does the job.

[watertrade]

just a thought...

from wikipedia.

Tindalization[1] /Tyndallization[2] named after John Tyndall is a lengthy process designed to reduce the level of activity of sporulating bacteria that are left by a simple boiling water method. The process involves boiling for a period (typically 20 minutes) at atmospheric pressure, cooling, incubating for a day, boiling, cooling, incubating for a day, boiling, cooling, incubating for a day, and finally boiling again. The three incubation periods are to allow heat-resistant spores surviving the previous boiling period to germinate to form the heat-sensitive vegetative (growing) stage, which can be killed by the next boiling step. This is effective because many spores are stimulated to grow by the heat shock.

I have heard of people using this technique for growing mushrooms.

[Teljkon]

Holy Pf tek batman my start up costs just went down considerbally.