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Can substrate and casing that never colonized be reused after...


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#1 Redrock35

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Posted 21 July 2020 - 08:30 AM

I have 4 substrate bags out of 6 that did not colonize and if I were making my own instead of spending $$$ for them, I would probably toss them with question. Can I sterilize them in a pressure cooker and bring it back to field consistency to be reused and does this have drawbacks from purchasing fresh bags? 100% of my jars colonize, but having little luck with bags. I've also lost 2 bags to the break a bag up with 1/2 colonization to make 100% of the bag colonize quicker. Not doing that again soon.

My other query is this, My A+ 100% colonized in the jars, I broke up the mycelium and cased it and waited 10 days to expecting see bits of white mycelium poking through the surface, no go. So I wait another 10 days looking in and misting, still no growth. I decided that after that amount of time, I added light and moisture to see if anything was going to pop up... but still nothing. Can this be reused after sterilization? 

 

If this is in the wrong area, please advise. Thanks for your time.

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Edited by Redrock35, 21 July 2020 - 10:10 AM.


#2 PJammer24

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Posted 21 July 2020 - 10:59 AM

Long ago, when I attempted to purchase bags like this, I had very little success... At the time, I suspected that they had been made in advance and after sitting for a time, were no longer adequately hydrated. You could potentially re-pasteurize, you pasteurize substrate rather than sterilize, but you would have to remove it from the bag and go from there... rehydrating it and pasteurizing in a pillowcase on the stove is possible... I steam pasteurize but that would require and investment and some setup on your part. I would suggest simply doing it on the stove in a pot of water with the substrate in a jersey knit or other porous pillow case... I have also used mesh laundry bags.



#3 coorsmikey

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Posted 21 July 2020 - 06:07 PM

I would be more concerned on why you have a substrate and casing laid out in a tub not colonizing than trying to repeat the same results. Why did they fail to colonize in the first place? Did you forget to add spawn. Did you try to short cut spawn and shoot a spore syringe directly to the substrate. There is something critical missing here that you should figure out before recycling or even investing in new bags.


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#4 Redrock35

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Posted 21 July 2020 - 08:05 PM

Long ago, when I attempted to purchase bags like this, I had very little success... At the time, I suspected that they had been made in advance and after sitting for a time, were no longer adequately hydrated. You could potentially re-pasteurize, you pasteurize substrate rather than sterilize, but you would have to remove it from the bag and go from there... rehydrating it and pasteurizing in a pillowcase on the stove is possible... I steam pasteurize but that would require and investment and some setup on your part. I would suggest simply doing it on the stove in a pot of water with the substrate in a jersey knit or other porous pillow case... I have also used mesh laundry bags.

Would a slow cooker work at 145 F for about 30 minutes?



#5 Redrock35

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Posted 21 July 2020 - 09:12 PM

I would be more concerned on why you have a substrate and casing laid out in a tub not colonizing than trying to repeat the same results. Why did they fail to colonize in the first place? Did you forget to add spawn. Did you try to short cut spawn and shoot a spore syringe directly to the substrate. There is something critical missing here that you should figure out before recycling or even investing in new bags.

You know, your right. I need to invest in a laminar flow hood, use more 70% isopropyl alcohol instead of 90%,(who knew less is more because of the water content) and even though I have put up a 4'x8' cleanroom grow tent, using  seedling heat mats and digital thermostat combos to keep temps exact, and a humidity control set up to keep it 75% to 90% and I'm now filtering more air that before, I still have a ways to go in equipment and knowledge. Jars work great for me, but they are a bit expensive in large numbers. (but now have lots of jars for future use) That casing has 9 of the 1/2 pint jars of A+ colonized mycelium layered into it, and in retrospect I should of used at least 12 for the 5 pounds of casing I used. I'm trying to step up my game, and the answers I get from reading and asking questions here is where I'm learning a lot of it from. I can't exactly pinpoint why the majority of my bags don't make it.  More than likely I've been overdoing it in the amount of spores (liquid) I'm injecting because of the opaque milky brownish liquid in the bottom of the bags. For sure I've lost 2 bags in separate incidents from breaking up the mycelium and reforming at 1/2 colonization to speed it up, which was extremely disappointing. I'm not exactly a patient guy, but I'm sticking to what I know works and pushing the envelope making my way slowly but surely up to mixing my own substrate, casing and cloning decent spores. 

 

Any advice you have would not only be welcomed, but my spores will likely thank you as well.


Edited by Redrock35, 21 July 2020 - 09:47 PM.


#6 TVCasualty

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Posted 22 July 2020 - 01:19 PM

Don't try to re-use bags of grain spawn if they don't take the first time. I wouldn't re-use bulk substrates if they failed to take, either (straw, manure).

 

FWIW, when I pasteurize straw or manure I aim for ~160℉ and try to keep it there for at least 6 hours. It ends up hovering between 155-165 as the heating element turns on and off. Most of the time I pasteurize for 12 hours (with the heat on, and I don't start the timer until everything has warmed up to 160) and it usually stays at pasteurization temp for another 4-5 hours (after I turn the heat off) and then takes another 10-12 hours to cool all the way down to where I can spawn it. You can find success with less time and a lower temp, but success is more consistent and the general results seem to be better with longer times and higher temps (maxing-out at ~170℉). I find the extra time and effort worth it.

 

One thing that will improve results with the grain bags is inoculating them with a liquid culture that you grew from spores rather than the spores directly.

 

You're lucky to have any success at all with shooting spores directly to grain since in most cases something else (usually bacteria) takes over before the spores sprout and the mycelium can gain a foothold. Speed is an important variable in growing fungi (there's a reason -several actually- why Stamets titled his last book Mycelium Running). Starting spores in a liquid culture (LC) and inoculating the grain with that instead of spores gives you a head start of about a week, which is easily enough time to make the difference between success and failure. Granted, it takes a week or so to grow out the LC, so you're not shortening the time it takes to do a grow. The "head start" refers to the amount of time a LC takes to colonize grain vs. straight spores.

 

Also, making LC's extends your spores exponentially, which is important when you're not making your own prints (yet). So with a few LC jars you can use as little as 1 cc to inoculate 4 LCs and they can in turn be used to inoculate several bags of grain each (the trade-off is between higher inoculation rates per bag vs. more bags, and higher rates mean faster colonization while more bags means more total volume if they all succeed). If your sterile technique is well-practiced and your location is really clean you can go for more bags in hopes of a higher total yield but if you're still getting everything dialed in and your grow space isn't ideal (most aren't) then going for faster colonization is the way to go to maximize your chance of success (your total yield will be lower, but 5 bags that successfully fruit is arguably preferable to 10 that don't).

 

It's also worth considering the possibility that growing strictly from cakes could be the best approach, depending on your situation. I used to grow exclusively from cakes made from wide-mouth pint jars, which can provide more mushrooms than most would assume and while making a bunch of jars took a lot of work, it also spared me many potential points of failure (no whole grain, no agar, no LC, no pasteurization) since I shot spores direct into BRF jars. I also didn't need a flow hood or glove box to grow "bulk" quantities (a dry pound or more). It required making my own spore syringes since it used a ton of spores, but that's pretty easy compared to mastering the use of bags of sterile grain spawn.


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#7 PJammer24

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Posted 22 July 2020 - 02:12 PM

reading TVcasualty's comment, I was under the impression that we were talking about bulk substrate that wasn't colonizing rather than grain spawn... I would also not suggest reusing grain spawn. You shouldn't need any of the fancy equipment and unless you are adding an entire syringe worth of spore solution, that shouldn't be the issue. Even if you use an entire syringe, the excess water shouldn't keep it from colonizing....

 

If we are talking about bags of substrate rather than bags of grain spawn, you should not be injecting the spore solution directly into it. You will not have good or even any results from putting the spore solution directly into a bag of substrate. You need to colonize grain spawn first and then mix the spawn into the substrate...

 

Just to help be sure we are on the same page...

Grain spawn = hydrated and sterilized grain that we inoculate with either a multi-spore solution or a liquid culture. You can also do grain to grain transfers,

                        add a wedge from an agar plate, etc...

 

Substrate = a semi-nutritious medium that is mixed with grain spawn and then fruited. This can include any number of materials including coir,

                   straw,   poo, verm, etc....

 

casing layer = non-nutritious layer on the surface of the substrate. This layer helps maintain moisture content and creates a                                          

                     micro-climate    that    promotes     fruiting. I rarely, if ever, use a casing layer for cubes when growing on bulk substrates.

                    I have never seen much of a benefit or superior results of any kind when applying a casing layer. It is not typically considered necessary with 

                   bulk substrates.

 

 

Cubes are really forgiving. You can achieve great results without a flow hood, in temperatures ranging from 67/68F to 76/77F, and with a wide RH range... You can achieve great results without a flowhood, I don't currently have one.

 

I break up the mycelium in every one of my spawn jars after approximately 15-20% colonization. When shaken, they will colonize more quickly...

 

To help you figure this out, we need to get some more details. I think there is a disconnect due to your not having the vocab quite right yet... You mention jars but then you also mention adding spore solution to your bags that are not colonizing. If you are using spawn jars, the spore solution would be going into the jar rather than the bag... Only once the jar has been fully colonized will you be mixing it with the substrate you purchased in bags. I would mix the spawn directly into the bag rather than moving it into a different receptacle. I would then roll the top of the bag, clip it, and throw it on the shelf... I use a 1:1 spawn to substrate ratio and get full colonization of the substrate in 2 weeks or less...

 

I will try to check back before I go home for the day... I do must of my Topia posting while at the office... See if you can clarify your use of the terms as I have mentioned... Describe your spawn type and whether it is in a jar or bag... Describe your substrate and if it is in a jar or a bag... Describe your casing layer...

 

fyi... you would not be layering a casing layer with anything. The casing layer, which is not needed for bulk substrate with cubensis, is a single layer that sits on the substrate's surface when the substrate is reaching full colonization... The spawn, once fully colonized, gets mixed in with the substrate and then you wait until it has been colonized...

 

If I was you, I wouldn't worry about casing layers, not right now at least... Just figure out your substrate and your spawn, mix them, and you should have good results


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#8 Redrock35

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Posted 22 July 2020 - 02:40 PM

Don't try to re-use bags of grain spawn if they don't take the first time. I wouldn't re-use bulk substrates if they failed to take, either (straw, manure).

 

FWIW, when I pasteurize straw or manure I aim for ~160℉ and try to keep it there for at least 6 hours. It ends up hovering between 155-165 as the heating element turns on and off. Most of the time I pasteurize for 12 hours (with the heat on, and I don't start the timer until everything has warmed up to 160) and it usually stays at pasteurization temp for another 4-5 hours (after I turn the heat off) and then takes another 10-12 hours to cool all the way down to where I can spawn it. You can find success with less time and a lower temp, but success is more consistent and the general results seem to be better with longer times and higher temps (maxing-out at ~170℉). I find the extra time and effort worth it.

 

One thing that will improve results with the grain bags is inoculating them with a liquid culture that you grew from spores rather than the spores directly.

 

You're lucky to have any success at all with shooting spores directly to grain since in most cases something else (usually bacteria) takes over before the spores sprout and the mycelium can gain a foothold. Speed is an important variable in growing fungi (there's a reason -several actually- why Stamets titled his last book Mycelium Running). Starting spores in a liquid culture (LC) and inoculating the grain with that instead of spores gives you a head start of about a week, which is easily enough time to make the difference between success and failure. Granted, it takes a week or so to grow out the LC, so you're not shortening the time it takes to do a grow. The "head start" refers to the amount of time a LC takes to colonize grain vs. straight spores.

 

Also, making LC's extends your spores exponentially, which is important when you're not making your own prints (yet). So with a few LC jars you can use as little as 1 cc to inoculate 4 LCs and they can in turn be used to inoculate several bags of grain each (the trade-off is between higher inoculation rates per bag vs. more bags, and higher rates mean faster colonization while more bags means more total volume if they all succeed). If your sterile technique is well-practiced and your location is really clean you can go for more bags in hopes of a higher total yield but if you're still getting everything dialed in and your grow space isn't ideal (most aren't) then going for faster colonization is the way to go to maximize your chance of success (your total yield will be lower, but 5 bags that successfully fruit is arguably preferable to 10 that don't).

 

It's also worth considering the possibility that growing strictly from cakes could be the best approach, depending on your situation. I used to grow exclusively from cakes made from wide-mouth pint jars, which can provide more mushrooms than most would assume and while making a bunch of jars took a lot of work, it also spared me many potential points of failure (no whole grain, no agar, no LC, no pasteurization) since I shot spores direct into BRF jars. I also didn't need a flow hood or glove box to grow "bulk" quantities (a dry pound or more). It required making my own spore syringes since it used a ton of spores, but that's pretty easy compared to mastering the use of bags of sterile grain spawn.

Thank you, I've got a Liquid Culture Kit - Easy Spore Germinating & Mushroom Cloning System coming Friday, I had considered this before when I ordered it, but wasn't 100% sure, but from the enlightening info you I feel confident now I'm on the path. Hands down the best advice I've gotten so far, likely because this time I provided enough information for a more in depth analysis, you have made my day TVCasualty, again I thank you. One the biggest reasons I started to go bigger is that the potency of my grow is awful, Im air drying and the using desiccant to get them cracker dry before coffee grinding to a fine powder then using 00 capsules. I've worked myself up from 1 to 6gms of APE and GT. I did a GT 9+ gm recently and barely got a lift and I checked to make sure I don't have a medication interaction. 2 to 4 gms used to unlock the secrets of the universe, now days I cant unlock the secrets of the garden shed! Don't know why I can't seem to peak, but it won't be from a lack of trying. Have you ever seen such poor potency before?

 

BTW, How long will LC stay viable and at what temp?


Edited by Redrock35, 22 July 2020 - 02:48 PM.


#9 Redrock35

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Posted 22 July 2020 - 05:38 PM

reading TVcasualty's comment, I was under the impression that we were talking about bulk substrate that wasn't colonizing rather than grain spawn... I would also not suggest reusing grain spawn. You shouldn't need any of the fancy equipment and unless you are adding an entire syringe worth of spore solution, that shouldn't be the issue. Even if you use an entire syringe, the excess water shouldn't keep it from colonizing....

 

If we are talking about bags of substrate rather than bags of grain spawn, you should not be injecting the spore solution directly into it. You will not have good or even any results from putting the spore solution directly into a bag of substrate. You need to colonize grain spawn first and then mix the spawn into the substrate...

 

Just to help be sure we are on the same page...

Grain spawn = hydrated and sterilized grain that we inoculate with either a multi-spore solution or a liquid culture. You can also do grain to grain transfers,

                        add a wedge from an agar plate, etc...

 

Substrate = a semi-nutritious medium that is mixed with grain spawn and then fruited. This can include any number of materials including coir,

                   straw,   poo, verm, etc....

 

casing layer = non-nutritious layer on the surface of the substrate. This layer helps maintain moisture content and creates a                                          

                     micro-climate    that    promotes     fruiting. I rarely, if ever, use a casing layer for cubes when growing on bulk substrates.

                    I have never seen much of a benefit or superior results of any kind when applying a casing layer. It is not typically considered necessary with 

                   bulk substrates.

 

 

Cubes are really forgiving. You can achieve great results without a flow hood, in temperatures ranging from 67/68F to 76/77F, and with a wide RH range... You can achieve great results without a flowhood, I don't currently have one.

 

I break up the mycelium in every one of my spawn jars after approximately 15-20% colonization. When shaken, they will colonize more quickly...

 

To help you figure this out, we need to get some more details. I think there is a disconnect due to your not having the vocab quite right yet... You mention jars but then you also mention adding spore solution to your bags that are not colonizing. If you are using spawn jars, the spore solution would be going into the jar rather than the bag... Only once the jar has been fully colonized will you be mixing it with the substrate you purchased in bags. I would mix the spawn directly into the bag rather than moving it into a different receptacle. I would then roll the top of the bag, clip it, and throw it on the shelf... I use a 1:1 spawn to substrate ratio and get full colonization of the substrate in 2 weeks or less...

 

I will try to check back before I go home for the day... I do must of my Topia posting while at the office... See if you can clarify your use of the terms as I have mentioned... Describe your spawn type and whether it is in a jar or bag... Describe your substrate and if it is in a jar or a bag... Describe your casing layer...

 

fyi... you would not be layering a casing layer with anything. The casing layer, which is not needed for bulk substrate with cubensis, is a single layer that sits on the substrate's surface when the substrate is reaching full colonization... The spawn, once fully colonized, gets mixed in with the substrate and then you wait until it has been colonized...

 

If I was you, I wouldn't worry about casing layers, not right now at least... Just figure out your substrate and your spawn, mix them, and you should have good results

I have jumbled several terms here lately, and the followed PDF & video methods recommended by the company I purchase supplies from, (hence the successful grows) I've made mistakes with quantity and adhesion to the included instructions (hence the fails) and I have had some success with automating to be here when I'm not. I should of posted separately for each type of grow, that added confusion too. My mind wonders sometimes and my dialogue can be hard to follow. I really appreciate your patience and your help and although I would like to go more in depth, my day has to be cut short. I'll have overnight hospital stay tonight, so I need to get my ducks in a row (getting older eventually sucks BTW) . Thanks again & have a great night.



#10 PJammer24

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Posted 23 July 2020 - 08:32 AM

@TVcasualty... What is your reasoning for not re-pasteurizing substrate? I don't typically have substrates that do not colonize,  but in the past, i have run short on time and eneded up repasteurizing and using it... I don't see any reason not to attempt to use old substrate materials. Spawn is a different story all together. If it doesn't colonize, I typically suspect bacteria, and I start over.



#11 TVCasualty

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Posted 23 July 2020 - 07:12 PM

It's probably more of a problem for me than most growers thanks to where I live, which has an extremely high endemic mold spore count in the air year-round. So I'm already starting at a serious disadvantage in the fight against contams and every little bit helps (or hurts).

 

If I pasteurize straw or manure but don't use it right away it will be getting a nice head start on growing other stuff before I can re-pasteurize it, and the success of pasteurization (or sterilization for that matter) is partly dependent upon how "dirty" the material is before pasteurizing or sterilizing it. I'd re-pasteurize straight coir however, since it's very slow to contaminate even without pasteurizing it first. But I don't use coir anymore.

 

 

I started getting much better and more-consistent results when I took extra care to only use the freshest, cleanest straw I could find vs. whatever I grabbed out of the back of the truck at Home Despot. The straw sold at hardware stores or a feed-and-seed was almost always moldy with lots of black splotches on it while the stuff I got at Tractor Supply that was sold inside the store wrapped in plastic was bright gold/yellow and had no sign of any funk on it at all (and also happened to be pre-chopped, which is awesome). That pre-chopped stuff has been the cleanest source of straw I've found anywhere so far, and I typically get 5-6 flushes from my 100% straw bulk subs with it if I let it them go that long. And I don't even have to chop it up anymore.

 

That said, I've only had to toss some pasteurized substrate on one occasion because I couldn't use it soon after pasteurizing, so my policy of tossing it is more about my being extra-careful about cleanliness because I live in a microbiological nightmare therefore the risk/reward ratio of re-pasteurizing vs. starting over with new material is not worth trying it in my case. But it would probably work okay for those who live in a less contamination-saturated environment, especially if using manure.

 

Some aspects of how I approach growing would probably seem over the top and excessive to people who live in more forgiving climates, but if they moved into my neighborhood they'd understand why I do things the way I do real fast. And I usually don't mention this when i post advice or answers since I'm basically posting the most conservative approach that's intended to work in the most difficult environments because if they work for me where I live, they'll work for anyone living outside of a tropical climate.


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#12 Redrock35

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Posted 24 July 2020 - 10:34 PM

It's probably more of a problem for me than most growers thanks to where I live, which has an extremely high endemic mold spore count in the air year-round. So I'm already starting at a serious disadvantage in the fight against contams and every little bit helps (or hurts).

 

If I pasteurize straw or manure but don't use it right away it will be getting a nice head start on growing other stuff before I can re-pasteurize it, and the success of pasteurization (or sterilization for that matter) is partly dependent upon how "dirty" the material is before pasteurizing or sterilizing it. I'd re-pasteurize straight coir however, since it's very slow to contaminate even without pasteurizing it first. But I don't use coir anymore.

 

 

I started getting much better and more-consistent results when I took extra care to only use the freshest, cleanest straw I could find vs. whatever I grabbed out of the back of the truck at Home Despot. The straw sold at hardware stores or a feed-and-seed was almost always moldy with lots of black splotches on it while the stuff I got at Tractor Supply that was sold inside the store wrapped in plastic was bright gold/yellow and had no sign of any funk on it at all (and also happened to be pre-chopped, which is awesome). That pre-chopped stuff has been the cleanest source of straw I've found anywhere so far, and I typically get 5-6 flushes from my 100% straw bulk subs with it if I let it them go that long. And I don't even have to chop it up anymore.

 

That said, I've only had to toss some pasteurized substrate on one occasion because I couldn't use it soon after pasteurizing, so my policy of tossing it is more about my being extra-careful about cleanliness because I live in a microbiological nightmare therefore the risk/reward ratio of re-pasteurizing vs. starting over with new material is not worth trying it in my case. But it would probably work okay for those who live in a less contamination-saturated environment, especially if using manure.

 

Some aspects of how I approach growing would probably seem over the top and excessive to people who live in more forgiving climates, but if they moved into my neighborhood they'd understand why I do things the way I do real fast. And I usually don't mention this when i post advice or answers since I'm basically posting the most conservative approach that's intended to work in the most difficult environments because if they work for me where I live, they'll work for anyone living outside of a tropical climate.

Seeing your particular environment having disadvantages, its got me thinking. There seems to be as many ways from spore to flush as there are watching or listening to Terence McKenna right now, so I probably need to brush up on the reason and mechanics of propagation, ideal environment for spores and substrate, and contamination. The high plains desert that surrounds me is quite opposite but not without it problems. Though I'm thinking it would be tougher there to fight contamination. I appreciate you help & enjoyed reading your reply, thank you.


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