02/26/14-03/05/14

On this day at Ecovative, I learned how to work the ECA machine. (Electro Chemical Activation technology). Using this machine, I prepared for and carried out chemical treatments for floral foams, a product that is currently in the workings at Ecovative. Put more simply, Electro Chemical Activation is used to make ECA solutions by "mixing readily available food grade salt with water thereafter passing the brine solution through patented reactors...once inside the reactor, the brine is activated by way on an electrical charge and two distinct solutions are produced."

Put in chemical terms:

H20 + NaCl <=> NaOH + HClO
(H+ + OH- + Na+ + Cl- <=> Na+ + OH- + H+ + ClO-)

This machine was used because normally mycelium produces Acly groups that are hydrophobic. However, because we were out to create floral foams (a spongy foam that soaks up water and acts both as a preservative to lengthen flower life and a support to hold them in place), we had to make the very groups hydrophilic. Thus, by treating it with ECA solutions, aka, by using a strong base to deacylate the group, we were, put in layman's terms, "chopping off" the hydrophobic part.

To the left is a diagram of an Acly group. R simply stands for any chemical chain. The hydrophobic part of acyl groups is the double bond between the carbon and the oxygen molecule. Consequently, by using the ECA machine, I deacylated the chain and "chopped" that specific part out to make the product hydrophobic.

The solution I am pouring out here is the solution made from the ECA machine that will be inserted and sealed with the raw floral foams (in bags) as shown in the bottom picture. This is so that the deacylate reaction takes place, making the floral foam products hydrophobic rather than hydrophilic. 

a more close up shot of the required amount of solution for each bag (which contained 4  raw floral foams products)

After inserting the solution into each of the bags, the next and final step was to seal the bags using the Vacmaster as shown above. 

02/19/14

 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.

01/12/14: Different Types of Mushroom Substrate

          This past Wednesday, I spent my time again in the "dirty room" aka the room where Ecovative keeps all of their substrates. A substrate is basically any substance on which mycelium will grow on. There is an endless number of waste products mushrooms can decompose and when cultivating mushrooms, based on the particular substrate one can obtain different characteristics. Each time I come to Ecovative, I, time and time again, become amazed at the fact that mycologists can literally use almost anything we think of as "trash/waste" and turn it into a viable product that, in a nutshell, can help save the Earth!!

For instance, mycologists use most of the major debris that comes from farming or forestry.

       Of course one can not simply pick a batch of twigs from the forest floor and start cultivating. Mycelium grows best on substrates when they have been broken down into smaller pieces, 1/4 to 2 inches. 

Some of the different types of materials that can be processed by mycologists include:

• bambo
• brewery waste
• cacao shells
• cacti
• coconut/ coconut husk fiber
• coffee beans, grounds, hills, & debris
• corn, corncobs, cornstalks
• cotton & cotton waste
• fabrics
• garden waste, grass clippings, & yard debris
• hair
• hemp
• leaves
• manure
• nut casings & seed hulls
• oils (vegetable & petroleum)
• paper products (newspapers, cardboard, money, & books)
• soybean roughage
• straws (wheat, rye, rice, oats, barley, etc)
• sugarcane
• tea, tea waste, leaves, & trimmings
• textiles
• tobacco & tobacco stalks
• trees, shrubs, brush, & wooden construction waste

Stamets, Paul. Mycelium Running: How Mushrooms Can Help save the World. Berkeley, CA: Ten Speed, 2005. Print.

In spirit of Valentines Weekend, here's a cute image for you!

http://cheezburger.com/7485785344

02/05/14

I could not go to Ecovative this Wednesday because we had a snow day.

(Also, this is a very general summary of the steps that is required to create the packaging product)

01/29/14

Last Wednesday at Ecovative I helped make bags in the dirty room. What I did that day is the very first step scientists here perform in order to create their final mushroom packaging product. The dirty room is the room that contains all myriad of compost materials used to create their mushroom packaging product. For instance, there is a container that holds different waste products like saw dust and chopped up corn stalks and husks. It was a bit like cooking—like following a set recipe. After I filled the required number of bags needed to be completed for that day with compost, I weighed out a specific amount of calcium carbonate, CaCO₃ and added it to each bag. This substance, CaCO₃ has the effect of enhancing the myclelial linear growth rate. After this step, I added about a half a liter of water into each bag and thoroughly mixed all the components together. These plastic bags and their contents are then sterilized in an autoclave for about an hour and then allowed to cool. Then the mixture is injected with small pellets of mycelium and left to grow. I may have explained this process once before, but this was my first time that I actually got to start from the very beginning, with nothing but the rare ingredients. The person that I worked with, Christina, my mentor’s partner, showed me the different results/physical and chemical characteristics that one would get from using different compostable materials. When with my mentor, Courtney I mainly worked with analyzing the many different products, today I had fun stepping out of the zone and getting my hands dirty.

This video above may help you gain a better picture of what I meant in my explanation above. 

01/22/14

It’s known that a tip-high gradient of Ca2+ plays a regulatory role in mycelial tip growth in fungi and so I wondered whether potassium would have the same direct results. Unfortunately, as stated in my previous blog post, I found that this was not the case—either because something went wrong during the process of my experiment or whether there is a scientific explanation. As I had predicted after seeing my results, according to “The Effect of Cations on the Growth of Fungi” by E.B.G. Jones and D.H. Jennings, “Sodium stimulates dry weight production at low concentrations but inhibits is at high concentrations.” This statement fits very well with my results as the set of plates that was inserted with a low concentration of potassium bicarbonate exhibited some growth as compared to the plate that was given an excess of the very cation. Consequently, in my opinion, it seems that the reason the set of plates with the greater amount of potassium bicarbonate did not grow at all is because if I had inserted too much of the substance into my agar mix.

While researching for this very explanation, I came across several other published papers concerning potassium bicarbonate and mycelium. More-so-than serving as a growth stimulator like Ca2+, it seems to be that KHCO3 is a very effective natural fungicide. It has been found that increased concentrations of potassium bicarbonate (KHCO3) can act as an alternative to synthetic fungicides. Thus, it was concluded that potassium bicarbonate was an alternative chemical agent for controlling in particular, a natural antagonist: R. solani AG 4 HG-I and S. sclerotiorum.

If I were a researcher at Ecovative that had the goal of researching about potassium bicarbonate and mycelium growth, I would continue to experiment with these two substances, trying to find the ideal percentage of the cation. This is what many scientists like my mentor do, in hopes of bettering their mushroom product for their consumers. However, I do not believe I will be continuing on with this project as my mentor has notified me that mold had started to form on the different plates as a result of my long stay away from the lab.

01/15/14: A New Year @ Ecovative!

This week, I went into Evocative and as usual continued to experiment on my own experiment. For those who do not remember what this was about, the purpose of this experiment is to determine the effect of potassium bicarbonate on the growth rate of the mycelium in plated agar culture. During my last visit, I made 3 sets of plates: one set was made with a high concentration of Potassium Bicarbonate (17.6 g/L), one was made with a low concentration of Potassium Bicarbonate (1.76 g/L) , and a control set (made with your every-day Potato Glucose Agar [PGA]). Thus, all combined, I made a total of 24 plates.

Upon measuring the plates, I have found that the mycelium placed within a medium made with a high concentration did not grow at all. In addition, I was also shocked to find that the mycelium placed within the control agar plates, in general, grew the most. Those in low concentration did how, but only ever so slightly. An added observation that I noted was that there was no mold on any of the plates—thus, I knew my results were not the effect of contamination. This made me come to the conclusion that it is highly possible that the amount of high concentration I used (17.6 g/L) inhibited the mycelium from growing at all. As of now, I do not have a clear reason why this is so and consequently, this calls for more research on my part which I am also excited about! I really love unexpected results like these and rather than being disappointed at my results, this unanticipated outcome is one of the very reasons why I love science and it’s factor of the unknown J!!

Starting from the top down and in the picture in the left (and from left to right in the picture above) are 2 high concentration, low concentration, and control plates.

Although due to the condensation that formed inside the plates the growth is not that visible, the results, as mentioned in my post were very surprising!