Hi all,
Firstly let me thank you for taking the time to read this all. It is a bit of a lengthy post, but in order to get the best help, I think a great deal of information is needed. So again thank you in advanced.
Its been awhile since I've been on this site, and I'm coming back because I'm having a problem. I ended up going to a music festival awhile back in June, and was gone for about five days. Ever since I've been having nothing but problems. I'll start at the top:
Before leaving for the festival I had one or two cakes left, mainly used up, so I threw them out. I was thinking that I would have ended up loosing them anyway, so I would just start over when I came home.
When I came home, I used up the remaining syringes I have had stored in a cardboard, takeout, box in the fridge. I saw this as a perfect storage box because it would keep the syringes in a dark area regardless of the fridge being opened and closed. I had enough spore syringes to knock up about eight jars. I followed all of the usual techniques for knocking up jars(normal BRF cake recipe; top, dry layer of verm; the heating of the syringe needle; you get the idea). I then placed them all in my incubator (two five gallon buckets; one inside the other, with a water heater in the bottom one). Now some of the jars got contaminated almost instantly. Some didn't take off at all. Of the eight, I think roughly 3 made it past the 50% colonization point. I lost one around around the 65%, another around 80%, and the last around the 95-98% mark. The contamination that I've experienced has ranged from the common trich, to black mold, to an off grey, to a rust/yellowish color. I don't have pictures as the camera on my phone is damaged and does not work.
After this loss, I made up some new syringes from some spore prints I have from roughly a year ago. I followed the techniques for making up syringes(boiling water and sucking up/squirting out the water 5 times, and holding the hot water inside the syringe until it cools to room temp, etc.). After making up the syringes, I knocked up my whole set of jars (around 48 or so). I again followed all of the above step and what have you. I got nearly the same exact results as the first batch with the colonization, the loss of cakes, and the mold. Today I lost my last cake, which prompted this post.
So my questions for the community are: What in the hell is going on? (I was thinking that if my spore print was contaminated that I would loose all of the jars right off of the bat. Is this incorrect?)
Can having my incubator on the ground increases my chances of contamination?
Would boiling all of my jars before trying again help?
Thanks for any advice in advance guys.
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There are many vectors of contamination in the situation you have described, I will describe a few of the problems I see.
1) A fridge is a nasty place full of mold and contams. Keeping syringes in the fridge is a bad idea, they will last for years at room temp. Keeping them in a takeout container is an even worse idea, was it clean atleast? lol
2) Incubators are a bad idea too, especially one with stagnant water. If the water was sitting in that bucket while you were away it is defiantly a contam zoo by now. Some people swear by them, but I think incubators are a bad idea. Most myc is happy at room temperature. A poorly constructed incubator (especially a closed environment with standing water) is a death sentence. The day or too you might gain during colonization isnt worth the risk.
3) On your second attempt the spore syringe you constructed is likely the culprit, but its hard to say because you only tested it on 1 jar.
It might be a silly question but you are doing all your work in a still air box right? What is your sterile procedure like?
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