February 19

One of my flow charts

Today, I was supposed to work with Sarah, but she was at an appointment.  So, I learned how to sort corn substrate.  The substrate is sorted by size, and it was done so by a vibrating machine of sorts.  There were holes of different sizes in the machine so that when the substrate passed by, the different sized particles would fall through the holes accordingly.

After sorting corn, I went to work on designing the procedure of my experiment.  From my research, I knew the general outline that I had to follow, but the details were vague.  I didn't know what equipment was available to me, and since I want to work with a pathogen, how was I going to do this without contaminating the whole lab?  Courtney clarified many of these details and suggested that I utilize a flow chart to help organize my thoughts and details.  The flow chart especially helps with figuring out the timing.  What else could I be doing when the mushrooms are drying or how long does it take to centrifuge?  Since I have limited time, this is crucial.

My final experiment will be comparing the antibacterial properties of Pleurotus ostreatus and Coprinus cinereus on Aspergiillus niger.  

My first DoE, the one looking to compare light and dark reactions came out with data.  Based off of the data for the fruiting bodies wet weight, the light mushrooms had the greatest average wet weight.  This is not what we predicted but it is helpful to know nonetheless.

February 12, C. cinereus

I worked with Courtney this time, and we spent time transferring plates (the first thing I did way back in October).  It was a good review of lab skills, especially involving contamination!  I had to remember not to open the pipette outside of the hood or let my hand pass over open containers (even if it is gloved and sprayed with alcohol).
I also made observations for the DoE on lighting regimes.  Here are some of the things I saw:

LIGHT SET
All 8 sets are fruiting
5 are in the beginning stages of fruiting
1 in the medium stage
1 in the middle/end stage, not quite ready for harvest
all 8 blocks fully colonized

DARK SET
All 8 sets are fruiting
Five are still in the early stages of fruiting
Two in the middle stages of fruiting
One nearly ready to be harvested

Next week the mushrooms should be ready to harvest.  Then I'll be able to weigh them for my data and write the rest of my DoE.

For my final experiment, I think I'm going to simplify my experiment by just comparing the antimicrobial effects of two mushrooms for the mold Aspergiillus niger.  One of these mushrooms for sure will be oyster mushrooms as we have a lot of them coming from Sarah's experiments for the self-growing mushroom product she is helping to develop.  The other mushroom might be one called Coprinus cinereus, also called the gray shag.  It is an edible mushroom.  This mushrooms short life cycle (two weeks in a lab) and easy cultivation makes it a useful organism to study and analyze genetics and molecular studies.  It has also been proven that this mushroom has antimicrobial properties against A. niger.

C. cinereus before spores are released.

C. cinereus after spores are released.

February 5!

Today was spent working with Sarah.  I helped out with preparing regrind and rehydrating blocks for other experiments apart of Sarah's coffee grind product for Back to the Roots Company.
My own experiment on lighting regimes for oyster mushrooms has come a long way.  I believe in a few weeks we will be able to harvest them.  Here are some pictures that summarize my experiment.

These are all part of the dark treatment.  A few blocks did not colonize, at least on the outside, such as the one in the middle.  However they did still grow mushrooms such as the one below.  From observation, the dark treatment blocks that did colonize seemed to colonize more. 

Dark Treatment, the largest of the oysters.  To answer a question Peggy asked during our meeting, we know when to harvest to mushrooms when they begin to produce spores.  This you can see, and it looks like fine dust.  Then we'll cut and weigh the mushrooms.

Dark treatment.  These mushrooms are nice.  

A light treatment.

These are baby mushrooms growing on the sides of the block that has no hole cut in it.  Since there is no air hole, these mushrooms will not last long, but as of this picture, they are trying to grow along the edge of the ziploc bag and up.
  

For my next experiment I'm still looking for possibilities.  Since this is really the first experiment I am designing myself, I don't want to complicate it too much.  Still thinking along the lines of mycofiltration, but I'm looking up pathogens.  Many of the pathogens in my book however were really dangerous, they trigger tuberculosis and cholera, but there was one that I might really be able to work with.  It is called Aspergillus niger.  This microbe causes black molds which is commonly found everywhere in the world.  Aspergillus niger has proven to be helpful in biotechnology and waste treatment but it can be harmful to human health if not treated carefully.  My mentors said they can also easily obtain this pathogen.

January 22 and 28: Dehydration, Rehydration, and Mycorestoration

On these two days, I alternated between working with Sarah and Courtney.  Both days I performed a variety of tasks.  Those that involved my project included dehydration and rehydration.  After these two processes, the mushrooms will begin to fruit and I will be able to collect wet weight data.  According to Sarah, all my mushrooms are doing well!  A handful of the light experiment blocks are contaminated with a mucor called Rhizopus which is also known as bread mold.  Luckily, this contamination is not too severe and  all my mushrooms are growing, healthy white.  Look forward to pictures in the next post!
When I was not working on my experiment, I helped perform other tasks such as making regrind for other projects (including one for Courtney involving engineered wood) and hydrating blocks for Sarah.

Both Sarah and Courtney gave me readings to begin looking at ideas for my own experiment.
In a book called Mycelium Running: How Mushrooms Can Help Save the World by Paul Stamets, I was introduced to the idea of mycorestoration.  This is a process by which fungi repair or restore the environment.

The following is a quick summary of four practices of mycorestoration:
Mycofiltration: filtering water, where the mushroom can act as a net to capture or digest toxins and contamination
Mycoforestry:  sustaining forest environments through an understanding of the mushroom's role in an ecosystem.
Mycoremediation: using mushrooms to break apart toxins and even heavy metals from the land.
Mycopesticides: Creating biopesticides from mushrooms

It would be interesting to pursue one of these topics, especially mycofiltration.  However, since these practices are new and much more needs to be discovered in these areas, this could make for a challenging experiment for me.  This just depends on what I want to focus on and the experiment purpose and design.

Here are some awesome mushrooms I discovered from the book Mushrooms of Northeast North America by George Barron for your viewing pleasure.

Panellus stipticus,a mushroom that looks ordinary by day but glows at night.

Marasmiellus candidus, they look like flowers.

Lycoperdon perlatum, Gem-Studded Puffball