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OfflineMonkeycyst
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Help! Just CANNOT get growth in my jars ..
    #4050210 - 04/13/05 02:18 PM (12 years, 16 days ago)

Greetings all. Any help will be welcome regarding my problems with getting growth in my jars. I have been trying for an eternity it seems to get my mushroom cultivation going the way it should. I will try to document my trials as clearly as possible to give you an outline that will hopefully give some of you an insite into where I may be going wrong. Up until now, I have hesitated posting this request for help possibly because I am stubborn but mainly because there is such a wealth of knowledge here already that SHOULD have provided me the know-how. For the life of me I just can`t work it out. Here we go...
Maybe starting with where I am now and then going backwards will be best. This is how I`ve been trying:
Cubensis spores ordered from Ryche(Mexican/Costa Rican) .. Made my own spore syringes following the standard shot glass technique making sure to allow the sterile mineral water to cool for a couple of hours in the syringes before sucking up the spores to avoid killing the spores. The fact that I have never had ANY contaminations of cakes is perhaps evidence of the lengths I go to to keep everything sterile. I can`t see anything wrong will the spore syringe-making process.
I`ll move on .. I`ve been sure to allow the spores to hydrate, allowing the syringes to sit for at least 48 hours on all occasions before using for inoculation. Here is my cake prep rundown .. Now, I live in an asian country were English isn`t the used language but have managed to get what I think is the correct ingredients for the basic cake. I`ve kept it simple using flour/vermiculite/mineral water mixed to the recommended amounts. First I hade trouble finding brown rice flour and tried rye flour with no success. Then I found some organic brown rice and bought a coffee grinder to make my own brown rice flour to eliminate the problem of additives ..still with no success. Should be noted here that I have experiment endlessly with slightly varing water content. Using half pint canning jars, I sterilize them using the boiling method for one hour (I don`t have a pressure cooker.. but again, I have never had a problem with contaminations). At times, I had water getting into the jars while boiling and ended up with slushy cakes so to avoid this, I used electricians` tape on both the holes in the lid AND around the rim of the lid then wrapped the whole jar in foil before boiling. After that I seemed to have avoided the problem of water getting in during sterilization. OK, the sterilization is done.. contents of the jar are looking good seeming to have a nice water content with some slight condensation on the inside of the jars. Now the jars are left over night to cool. Pretty good so far, right!?
Moving on to inoculation. Wiping the needle with alcohol then slightly flaming, again wiping with alcohol, the jars are inoculated. Again, I have experimented with varing amounts at each of the five nail holes(one in the centre) from 1/4 cc to almost 1 cc in each hole. I have also tried re-taping the holes after inoculation AND leaving the holes untaped before putting the jars in the incubation chamber.
OK, the incubation chamber.. The set-up I have made leaves the least reason for failure in my opinion since it really is great. Inside a big wooden chest with a lid that seals quite well allowing no light to enter (but air to flow in and out) I have a large plastic tub with no top. Into that plastic tub I half fill with water and maintain the temperature using a fish tank heater at 80-85 degrees F. I`ve played around with the temperature a little here too. Into that water bath I put another medium-sized plastic container that holds the jars and thermometer(placed in the centre of the jars). It has a lid but I have also tried making holes in it to allow for better gas exchange AND keeping it fully closed. Of course that container floats on the water bath so I have it tied down to make it half submerged. Then the wooden chest is closed. Sounds good doesn`t it? I have waited more than two weeks at this optimum temperature to see next to no growth. There have been a few occasions when I have seen mycelium growth after about 3 days that continues for a day or two then stops completely. Having done some reading in The Shroomery, reasons for halted growth seem to be either the wrong water content or lack of gas exchange. With that in mind I have experimented with those two variables .. still with no success at getting a cake to fully colonize. MAN, WHAT A NIGHTMARE !!
Doesn`t it sound like I`ve done my homework, been careful to keep the process sterile, and dealt with possible faults??
Going back a bit, here are some results.. I tried using steeped bird seed combined with verm and BRF. This was BEFORE I realized that grains are a big no-no WITHOUT a pressure cooker.. BUT I did get a couple of jars to half colonize before contamination set in. I ended up combining the colonized parts form the two jars together to make one complete cake, put it in the fruiting chamber and actually grew some shrooms! That has been the extent of my success to date. Now I`m contemplating getting a pressure cooker and trying bird seed again.
Taking into account the fact that my process seems good, I can`t help but think that either the vermiculite or the flour is no good. But how? My BRF is home made, the vermiculite IS vermiculite after all.. it says VERMICULITE on the packet!? I`m also thinking of trying the POPCORN method since it seems to avoid any problems associated with verm/BRF..
Seeing next to no growth at optimum temperature also makes me think the spore solution is no good. I`m very careful here though. One question though.. I use cleaning alcohol to wipe the inside of the shot glass before scraping the spopes into it but always allow the alcohol to evaporate first. I have wondered if any residual alcohol in the shot glass might kill the spores. Is this possible and could it be that I`m using the wrong alcohol?
Well that`s the rundown of my attempts to date. Popcorn looks like my next chance.. I don`t know.
Everyone, any advice, suggestions, or recommendations will be really really REALLY appreciated! Apologies for the length of this post but I wanted to make the best possible chance for getting some help. It really is quite sad that I should have such little success after the amount of effort I have put into it. I deserve better!
Thank you all. I look forward in anticipation to ANY replies..
Peace.


--------------------
"Worry is preposterous. We don't know enough to worry."


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InvisibleOJK
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Re: Help! Just CANNOT get growth in my jars .. [Re: Monkeycyst]
    #4051346 - 04/13/05 06:25 PM (12 years, 15 days ago)

Firstly, let me congratulate you on your excellent research! You display a depth of knowledge that is admirable, and have obviously approached this challenge in a methodical and well-informed manner.

While I have no definitive answer for you, I'll try to run through a few factors that might be effecting your grow.

1) Spores: making one's own spore syringes from a spore print can be difficult. While you have had no problems with contamination, perhaps you aren't using a large enough section of the print for each syringe?

Additionally, as you live in Asia, the spores you order will be shipped by air-mail, and possibly irradiated during transit or customs! That would explain your low germination rate.

Possible fixes for these problems might be to order a spore syringe and use higher volumes of spore solution to try to encourage germination.

Alternatively, maybe try to find some kind soul from your own country or a nearby country to send you a print. Maybe post in the trade forum?

Perhaps to ensure that your spores are germinating properly, you could try a liquid mycelium culture. It's not the easties technique in the world, but it would at least let you know if your spores are alright or not.

2) Materials: you mention that you are using organic brown rice. However, does perhaps your vermiculite contain some kind of fungicide? As it is used in horticulture, this isn't impossible. Take a look at the packet, see if it's got anything in it that it shouldn't.

3) Method: well actually, everything you've posted about your method seems absolutely fine. Just for the sake of completeness though, just make sure that at every stage, your substrate is light and not too water heavy, your spores or substrate never get too hot, and there is always adequate gas exchange. Are you compressing the substrate into the canning jars too densely? (the jars should be loose and full of air) Is your vermiculite filter layer too dense? Is your incubation box aerated enough?

As I say, nothing in your post leads me to believe that your doing anything like this wrongly, I mention it merely just in case.

Anyway, that's about everything I can suggest. I wish you the very best of luck, because it seems you've done alot of research and are deserve a great deal of success.

Good luck!


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OfflineAnnoA
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Re: Help! Just CANNOT get growth in my jars .. [Re: Monkeycyst]
    #4051361 - 04/13/05 06:32 PM (12 years, 15 days ago)

I know you said you didn't have contamination, but do the jars develop any unpleasant odour when colonizing?


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OfflineAbermelin
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Re: Help! Just CANNOT get growth in my jars .. [Re: Anno]
    #4051956 - 04/13/05 09:20 PM (12 years, 15 days ago)

Quote:

The fact that I have never had ANY contaminations of cakes is perhaps evidence of the lengths I go to to keep everything sterile.




( i didnt read the post entirely, so correct me if im wrong )

how can you claim to never have contams when you've never had ANY growth at all? A common strain of bacteria called baccilus is what keeps cakes from growing, or at least this was my problem for a whole year before i knew that contams can be invisible and prevent growth.

as far as dealing with fungicides goes... call the manufacter of whatever products your using ( if it says organic, then dont worry bout it) and ask if they put any fungicides on their product. most of the time they will tell you.

oh, and you NEED A PRESSURE COOKER! boiling doesnt do shit. so your in china eh? isnt most of the shit we use come from over there? im sure you could ask a little 6yr old to make one for you, oh, and ask them if they can make me a new pair of shoes, ill pay em $1 in labor.


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OfflineMonkeycyst
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Re: Help! Just CANNOT get growth in my jars .. [Re: OJK]
    #4053781 - 04/14/05 10:22 AM (12 years, 15 days ago)

Odiumjunkie.. Thanks a lot for taking the time to read my post and add your valuable thoughts. Here are my reponses to your suggestions:

When I`ve made my spore syringes I think I`ve always been quite generous with the AMOUNT OF SPORES I scrape into the shot glass. As an example, for a medium sized print, I would use between a third and a half of the print for one syringe. From what I know, this seems plenty, does it not?
What you said about the possibility of my ordered prints getting IRRADIATED when going through customs or the like scared me. That`s possible I guess and would explain the little to no growth. How does that happen?.. I mean, what process would irradiate the spores on their journey to my mail box? I`ve never heard of that .. just curious to know. ORDERING A SPORE SYRINGE would be better? Wouldn`t the syringes go through the same process that would render the spores worthless(irradiation)? Does the fact that the spores are in solution prevent that happening? I have always ordered spores through Ryche who will not post syringes to this country. I`ve never ordered from anyone else but I imagine there is a good reason for not posting them here. You`re right, getting spores from someone local would be the deal. May be hard but will look into it..
Your suggestion to try a LIQUID MYCELIUM CULTURE sounds great and provided some real hope for me. That would be a sure way to find out whether or not the spore prints I have are viable or not. Great idea! I`m off to the home brew store tomorrow.
With regard to the LIGHTNESS OF THE SUBSTRATE, even though I`ve had so much trouble, I think I have become somewhat of an expert mixer with all the practice I`ve had to date. I`m very careful with the water content- Never too sloppy/slushy nor dry.. it is moist without being too heavy. Always well mixed and `fluffed up` the best I can before loading into jars. And, I have never PACKED the substrate into the jars. I basically let the substrate drop into the jar from a spoon. DEPTH OF VERMICULITE LAYER is about half an inch. In addition, I have experimented extensively with all the variables.
The one point you made that sounds the closest to solving this painful riddle is that concerning the possibility of the VERMICULITE CONTAINING PESTICIDE. Because I have had(on a very few occasions) a little mycelium growth in the jars that halts completely very soon, I`m picking that the spores are fine but are killed by something in the substrate- FUNGICIDE! It makes sense. I can`t read the characters on the package but I can get a friend to do so. In the case of the vermiculite containing fungicide, I wonder if thoroughly washing the vermiculite before using it to mix up a substrate would help. That sounds troublesome and probably not so successful in getting rid of it.. but worth a try? Guess I could order verm from another country.. but first I`ll get someone to read the package of the stuff I have now. That would be a good start. I`m thinking, to eliminate that concern(fungicide), I could try the POPCORN TECHNIQUE which doesn`t require the use of vermiculite. In fact, I bought some popcorn tonight..
OK, wrapping it up.. Going by what I know and feel in conjunction with your wise thoughts, it appears quite clearly that the problem can be narrowed down to one of TWO probable factors. 1). My spore syringe is crap, or 2). Fungicide on the vermiculite is choking any attempt by the mycelium to grow. Sound good? With this in mind I`ll try your suggested liquid mycelium culture technique to find out whether the spores I have are good AND have the writing on the vermiculite package translated. May put down a couple of jars of popcorn substrate jars as well.
Thanks again Odiumjunkie.. really do appreciate your help. I feel I`m closer to sorting out this mystery. Any further ideas welcomed. Take it easy.. talk to you again soon I hope. I `ll keep you informed..
Peace. The Cyst.


--------------------
"Worry is preposterous. We don't know enough to worry."


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Invisibletripndicular
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Registered: 08/25/02
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Re: Help! Just CANNOT get growth in my jars .. [Re: Monkeycyst]
    #4053959 - 04/14/05 11:16 AM (12 years, 15 days ago)

Welcome aboard first off.  :thumbup: :cool:
And like ANNO asked have you noticed any foul odors coming from the jars ?

This would tell us if you are battling comtams or flat out stalling jars.
If you are getting germination and some growth this rules out the theory of not enough spores in the syringes. Try buying a pre made syringe and see if it does any better, that would tell you if it is your technique making them (I doubt that , not as tough as many make it sound ).
Also 85*F for colonization it a tad hi imho, but not deadly, but it can and will slow growth down considerably in many cases.
Now the point you made about "fungicides" doubtful but possible , make damn certain that none of the products you are using have this dastardly product in them , because it will stop growth dead in its tracks , but odds are that is not the case , otherwise you most likely would never see any myc growth at all. As far as spores being radiated by the postal system and customs , I doubt very seriously that is what is happening. I have received spores that I know for a fact were radiated and they were just fine.
Well hope you find your answers ....
OH YEAH PATIENCE is the key to successful mush growing , do not give up , in time you will find the answers and get many grows under your belt and this will all just add to your exp. GL
"May The Mushroom Goddess Bless All Your Endeavors!"

ps Not that it matters a whole lot , use "distilled" sterile water for making syringes instead of mineral water .Perhaps mineral content is a problem for you.


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


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OfflineMonkeycyst
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Registered: 09/27/04
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Re: Help! Just CANNOT get growth in my jars .. [Re: tripndicular]
    #4054081 - 04/14/05 11:57 AM (12 years, 15 days ago)

Thanks Tripndicular.

Foul odors? Well havimg aborting jars that haven`t shown any growth after 2 weeks, the substrate smells. I wouldn`t say foul, slightly pungent I guess. But I imagine anything organic and moist left for that long would start to smell. ??


--------------------
"Worry is preposterous. We don't know enough to worry."


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OfflineMonkeycyst
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Re: Help! Just CANNOT get growth in my jars .. [Re: tripndicular]
    #4054097 - 04/14/05 12:02 PM (12 years, 15 days ago)

.. I`ve been using boiled mineral water for my syringes. I`ve been using what I thought is the best quality- Volvic. Mineral content might be a problem? And, distilled water.. could I get that from a drug store? And why is that preferred over mineral water?


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"Worry is preposterous. We don't know enough to worry."


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Amazon Shop for: ½ Pint Jars, Brown Rice Flour, Herb Grinder, Pressure Cooker, Rye Grain, Vermiculite, Wild Bird Seed

Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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