Today we took the sclerotia out of the incubator. It didn’t look very promising. Most of it had died and looked infected. We cut the small area of ‘normal’ looking sclerotia out and kept in in a petri dish in hopes it would dry out completely and be usable in the future. We also changed the oats in our living culture.
Please see my inquiry partner’s blog entry for more details: http://annicah.inquiryhub.org/2013/03/05/march-4th/
On Thursday we checked on the progress of making sclerotia and transferred the filter paper from nutrient agar to empty petri dishes.
Yesterday we practiced our presentation and presented our project.
Today we worked on our PowerPoint presentation and pretty much finished it. We also plated petri dishes with nutrient agar and started the sclerotia process again. We are converting all our cultures except for one (the new one we cultured from our maze experiment) into sclerotia. The cultures (2 petri dishes of soon to be sclerotia) are now in the incubator and we will need to take them out about 24 hours from now.
Today we started a PowerPoint presentation for our inquiry project. Some of the dying/infected cultures are in the freezer – maybe we can bleach them tomorrow (to kill them).
When I checked on the cultures on Monday, the two that were dying looked like they had a bit of hope. I saw a bit of yellow slime mould in each, but one of them we overrun by another kind of mould. We did our best to scrape out the other mould and changed the oats in both cultures.
The slime mould finished solving the maze last night at around 7:30 pm. I got a picture of it at 4 pm, then forgot about it until 7:25 pm, when I checked on it and found that it had solved the maze, so I’m not sure exactly when it finished, but it was sometime just before 7:30 – maybe 6:30 or 7:00.
Today we transferred the filter paper from our second sclerotia attempt to an empty petri dish, cutting out sections that had other mould growing on them. We put that culture back into the incubator (along with the others) to let the filter paper dry out. We also changed the oats in the other living cultures.
Today at 9:24 am we put the slime mould into the maze! We took a small piece of it with the agar it was on (from one of our first sub-cultures) and placed it upside down at the start of the maze. We put several oat flakes at the end of the maze. After watching it and taking photographs periodically every half hour, it didn’t look like it was growing much so we gave it an extra oat flake at 12:50 pm, thinking maybe it didn’t have enough food to be able to get all the way to the finish. It has grown minimally since this morning (less than a centimeter in the last 5 hours). Considering the top growing speed we read about online (1-1.8 cm/hr.), we are wondering if a) it didn’t have enough food or b) we didn’t put enough slime mould in to start with. My partner is planning to take it home and keep photographing it as time progresses. Hopefully it won’t grow too much during the night. The maze is quite large in comparison to the organism, so it could take a while.
We have been keeping the rest of the cultures in the incubator as they seem to grow better in there, and have started the process of making sclerotia again with our original culture. We changed the oat flakes in what was left of the original culture and put it back in the incubator to keep warm.
After discovering that a couple of our cultures didn’t look like they were doing to well this morning, we decided to put off the maze test until Tuesday. 2 of the cultures appear to have retracted their veins and formed small crusty balls near the edges of the petri dishes. Perhaps the clean-up job we did yesterday was a bit too much for them, but in any case, we want to allow the cultures the weekend to grow and recuperate before putting them into the maze we created.
Today we started the process of reviving some of our sclerotia. We selected a petri dish prepared with non-nutrient agar, sprinkled a few oat flakes on it, added a piece of dried filter paper containing sclerotia, and applied a drop of water to the filter paper. We sealed and labelled the petri dish, then covered it in foil to keep it dark.
The slime mould kit we ordered arrived on Wednesday afternoon, but we are sill awaiting the incubator and other supplies. The mould will be left in the fridge for the weekend, and hopefully on Monday the incubator will arrive and we can start growing the mould. Once the culture is large enough, we hope to subculture it, and that way if one plate dies or becomes contaminated, we’ll still have some to work with. First we’re going to experiment with growing the mould, keeping it alive, sub-culturing it, and transferring it to and from its dormant stage. Then we’re going to try to work out how to set up the maze so we can test how long it takes to get through it.