Last Wednesday, for the first time in quite some time, I did not work in the "dirty room" but rather spent my time working in the lab. The week before I had made the compost and sterilized each of the bags. This week, consequently, I took these same bags and proceeded with the next step, which is to take apart the fully-mixed substrate which had been "glued" together by the ingrown mycelium. A new thing that I had not previously known was that after completely separating the substrate into bits and pieces, a certain tailored amount of flour was added to the mix. The flour was said to help speed colonization once the substrate we took apart were inserted into packaging molds.
The material I was working with last Wednesday and the week before was twine. My mentor and I found this to be a very tricky material to work the week before as we have to laboriously take apart every single clump of twine before inserting it into the bag of substrate we were making. Sometimes we would need something like 35grams of twine and that was a pain. With other materials such as corn husk, all I would have to do is measure out 35grams of the material, but with twine, I had to carefully pick out and come up with 35 grams worth of pieces (or take apart the lumps) of twine that were not clumped together.
The difficulty with twine did not stop from there. This week, my mentor and I found that of the 7 bags that we had painstakingly put together, about 3 to 4 of them had formed mold. Thus, long story short, we were not able to use them and they had to go to the trash. (such a waste, my heart broke upon throwing of throwing the very bags that I had spent so much time working on). My mentor explained to me that the very bags most likely became contaminated due to the process in which they handle the material. After we had made the bags and put them in the heat chambers for sterilization, the next step was to take apart the substrate in the bags and give them a thorough mix via gloved hands. This is when she suspects the substrate became contaminated. (due to the bacteria in the air--despite it being done in the sterilized lab). She informed me that this had to be done given the particular characteristics of twine. Unless someone comes up with a different way, this was, unfortunately, the protocol when dealing with twine. This incident had me realize yet again how susceptible materials are to contamination and consequently how important it is to be sterile and take that extra precaution no matter how silly it may seem. It also made me realize that there are times when the whole batch prepared a week before becomes contaminated and when scientists have to start again from ground 0.
I can imagine how upsetting this was to see your bags mold! The process you described seemed so meticulous, and I can only imagine you felt thought about all the time you spent on those bags. It is true, however, that what you learned from the experience is valuable. I'm interested to hear amore about how this relates to your own project? Do you think this experience will affect it?
ReplyDeleteThis is a different result of sterile technique. You have learned how to inoculate with specific strains, and now you have to deal with a persistent infection! Both situations are challenging, but this one seems especially so.
ReplyDeleteAs noted before, please add a few sentences about how your weekly work fits into the larger aspect of your project. This will help you refine your ideas, and it will allow your readers to follow along easily.
Christi, remember to blog every week, even if you miss a trip.
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