Due to the fact that all of our living creatures died prematurely, our group has decided to nix the mysid shrimp and midge fly assay and for the past few weeks have been spending all of our time working on the making the YES assay work. We have slightly altered the methods from the original protocol from Xenometrics, using centrifuge tubes instead of 96 well plates. In the beginning, our tests were working quite nicely and slowly but surely began producing odd results. Taking a step back and thinking about it, Dr. Wilcox realized it was most likely because we had been using old stevia AND old β-estradiol, which just happens to be air sensitive. So. Much to our dismay we will be redoing our tests after Christmas break and crossing our fingers in hope that we will end up with results before our first competition!
Due to reasons unbeknownst to us, the 100 Mysid shrimp that had been placed into premium condition habitats died. We assume that these reasons pertained to a lack of food and the nature of these beasts that leads to cannibalism. Duh! Duh! DUUUH!
On a side note, we began preliminary testing for the YES assay to see if our Stevia extract would kill the yeast and luckily, after letting the yeast grow in our extract with a growth medium, we had yeast! Yay!
Cameron, Erin, and Sam have officially started one of their 3 assays, using the Chironomus Riparius!As mentioned in the previous post “ Construction Zone” these midges are used to test agricultural chemicals. This is because their hormone that controls the larval stages, ecdysone, is so similar to estrogen that chemicals that activate estrogen receptors will also activate ecdysone receptors. This is what made these bugs such an assay for testing for estrogen in stevia!
After weeks of building cages, gathering dirt, and preparing habitats, our group is finally going to be able to start with testing by tomorrow, seeing as we will be getting our most important element of our project; Stevia. Hopefully, now that our prep work (for the most part) is done, fly cages being the main exception, we will be able to dry down our plants, and make extracts to prepare for our assays.
Cameron, Erin, and Sam are now building Breeding cages. This is part of an assay to test for the presence endocrine disrupting chemicals using Chironomus riparius, a non-biting midge. The hardest part is making sure the sides are completely closed, as these midges can be as small as 1 mm! They used these midges specifically because they are used very often while testing endocrine disrupting effects of commercial agricultural chemicals.
Cameron, Erin, and Sam have started to create the intricate set up that will be needed to find any Endocrine Disrupting Chemicals in many different extracts of Stevia and it’s compounds. Since Stevia is such a large player in the sweetener world it was quite confusing as to why the whole plant was be banned by the FDA… for many years. The student researchers are trying out three different tests that are all very time consuming and tedious, and are very excited to get started! They recently got the news that the Swiss company Xenometrix is giving them a $1600 kit for nothing other than shipping costs, to help them jump start one of their assays. With the news of really being able to get started these three are looking forward to being able to solve this puzzle once and for all.