Trials Have Begun! ... Several Have.
- KatyAnn Dudley
- Jul 7, 2017
- 8 min read
Updated: May 15, 2020
I haven't written anything about the trials yet, but this is the third trial I've completed.
Before we get into data, I'll show you what I used for containment units in these trials. In case some of you were worried I'd be conducting experiments on entire hives... I'm not! :)
I used a containment unit idea I got from this published paper:
Evans, Jay D., et al. "Bee cups: single-use cages for honey bee experiments." Journal of apicultural research 48.4 (2009): 300-302.
Here's a link to read it online if you want direct access to the article: http://www.tandfonline.com/doi/pdf/10.1080/00218839.2009.11101548
My method was modified however. I had to use 16 oz cups because apparently none of the stores here have 14 oz clear plastic cups... none.
So, since 16 oz cups were all I had to work with, I also couldn't use the base of a Petri dish to make the bottom of the containment unit. These lucky people who got a hold of a 14 oz cup (the authors of the paper), could use the base of a Petri dish (the smaller side) and it fit right over the rim of their 14 oz cup, kind of like a lid.
As you can see from the paper, they turned these cups upside down. Before nestling the cups within the Petri dishes, they placed a layer of incontinence pad between the Petri dish and the rim of the cup. Not only would this keep the entire unit secure, it would also keep the liquid in the scent vial from making the Petri dish below slippery or sticky (since they were using sugar water, it would end up being really sticky).
You can skim through the rest of that paper if you want to know more about how they put it all together, but I stated those specific parts because mine had to be a bit different.
Since, I couldn't use the smaller side of the Petri dishes, I had to use the larger lid. The problem continued with these 16 oz cups, however, because even the lid of the Petri dishes fit inside the rim of the cup, which isn't exactly ideal... but it worked so I was willing to go for it.
Instead of using incontinence pads, I bought paper towels... because they're cheaper. I then wrapped the Petri dish lids in paper towels and taped them with lab tape.
Below are pictures of step by step what I did to accomplish this:
P.S. I'm sorry this photo section is so long
Initially, when I had a "Trial phase" set up, I was comparing nurse bees with foragers. Nurses are one of the youngest work forces within a hive, so I hypothesized they would live longer than the foragers. I was correct in my "Trial", but in order to get nurse bees, I have to access the part of the hive where they're actively taking care of larvae.
In my current hive set up, I have a deep, another deep, a queen-excluder, and then a super. (Although, neither hive is using the super yet.)
My hives are technically still "small" so the only place they're currently taking care of larvae is in the very bottom deep box, but there are a lot of bees in the second deep, and on some unlucky chance: the queen. So, I really don't want to have to move that upper deep box off into the grass while I'm taking nurses from the lower deep for 45+ minutes.
So, instead of taking out nurses, I left the top deep on and just took whatever bees were crawling on the top bar of the frames in that top deep box.
Admittedly, this creates a lot of potential variability in my data collection because a forager from the neighboring hive may come in for a landing and that could be the one I snag. So, while I try to be as careful as possible and only grab bees I know come out of whatever hive I am working with at the time, there is that potential for error. Also, since I'm not specifically taking out nurses, now I could be getting workers and foragers in each cup. It's not really possible to tell the difference between a worker and a forager when everyone is crawling around on the top of a frame, so this may also account for some of the early mortality as well as the number of mortalities seen in the Control group.
For anyone who isn't aware:
Mortality: the quality or state of being mortal. Example: Her husband's death reminded her of her own mortality.
So, really, it's a fancy word for referencing to the potential for death of any living being you're discussing. In this case, it's bees.
So! Getting in to the more technical side of things:
For the first two experiments I had 40 total cups with 5 bees in each cup.
Experiment 1: Copper Sulfate, Concentrations: 0, .1, 1, 10, & 100
Experiment 2: Copper Sulfate, Concentrations: 0, 1, 10, 50, & 100
Copper Sulfate in solid form:
For the third experiment, we eliminated one of the concentrations so there were only 24 cups with 5 bees in each.
Experiment 3: Zinc Sulfate, Concentrations: 0, 1, 10, & 100
Zinc Sulfate in solid form:
To put the bees in the cups, I used round-tipped tissue forceps like the ones below:
There are many different types of forceps. For these trials I thought these would be the easiest to use and have the highest potential to cause the least amount of damage to the bees.
For the first trial I tried one method of capture that for the subsequent trials I decided I didn't think was the optimal method.
In trial one, I used the forceps to grab at the very base of a bee's wing, right along their back.
Previously, this arrow indicates where I would pick up the bees to put them in the cups. I never tore a wing off this way, but it did, from what I now know, caused more damage and it was more difficult to get the bees into the cups because they would use their legs and other wing to try and grasp at something else to get away.
In order to get them into the cups this way, it made me force them in a bit more harshly and sometimes injure another part of their body as well as their wing.
I hadn't known better at the time, since I was still nervous and wasn't sure what worked and what didn't yet.
My second trial went much better. Instead of grabbing the pivoting, and probably really painful, wing, I went for sliding the forceps along the top of the frame until I could secure two or three of their legs as shown here:
This was far less painful looking, but it was also really helpful in more than one way. The bees, when being seized by the legs, would curl up into tight little balls in an attempt to sting the forceps. This tight angry ball of fury was far easier to work with than the previous method.
This, also I think had a hand to play in reducing the amount of control mortality in my second trial.
I have just begun my third trial today with the Zinc Sulfate as mentioned previously.
Fun Fact: not a lot of people really think about it, but yes, bees do defecate. It seems to be similar to how birds poo, how the liquid and solid is all put into one disgusting concoction... except in the case of bees, it's as bright yellow as they are.
And now... more pictures and a harrowing tale about how I got them all back to the lab...
Oh! but first: These first two are of my first Nurse Vs Forager trial. Left is their first day in the containers, you can see they're drinking from the vial at the top. The one on the right is a little vial I put them into after they died. If anyone is wondering whether a bee you see sitting still is dead or not, if the legs are curled up underneath it at a sharp angle (kind of like how spiders die), the tongue is sticking out, then you know it's dead. The front two-four legs are the most common ones that are curled up underneath. The back legs may not.
Note; Any of the strange dark marks you see on my hands are probably from Sharpie®. Like that dot on my finger below.
^ That guy in the background is Joel, one of the other grad students that works in Scott's lab. I've talked to him once(?) and he's worked here for almost a year. He works with snails. That's all I know about him.
Hey... most scientist people just kinda do their work and talk when they have to, so don't look at me like that. It's almost considered rude sometimes when you're working in the same lab to interrupt someone's music (he always has his ear buds in... respect the ear buds guys) and their time doing their work, so yeah... I'm asocial, but I'm not being rude by not saying hello.
Below: This is what the final set up looks like, when they're all just sitting there... being slowly poisoned... for the greater good! For the contribution of science!
As a side note... I am really not fond that my research right now is in toxicology. I don't like killing things in the name of science. I understand why it has to be done, but I don't like it. Even insects... The goal here is that, by understanding their tolerance thresholds for what they're exposed to in the environment, we can use this data to keep more of them alive in the future. It's the in-between part of seeing how long it takes them to die after ingesting heavy metals that isn't fun.
Initially, transporting these guys back from my professor's house to the lab, I placed them how you see them in the picture above, with the lid side down. Unfortunately, on the bumpy car ride, it caused the liquid inside the vials to fall out and soak the paper towels as well as the bees below. In some really bad cases, the car ride caused over half the vial to empty into the paper towel below.
Much to my dismay, I couldn't really think of a better way to transport them with the materials I had available to me. It did, however, make sense that next time I should place them upside down so the vial wouldn't shake its contents out onto the bees below... but how to do that?
I tried wrapping each cup inside a paper box with this stuff:
but that was an absolute nightmare...
I was way too tired that morning to be patient with figuring how how to wrap them so well they all finally stayed upright on those tiny little glass vials.
Every time I do this trial, I start at around 8:00 and end somewhere around 16:00. So... at 8 that morning, I was exhausted from staying up the previous night to also do physics homework and finally in my distressed state and with a sad-and-frustrated-puppy-like expression I glanced to my right and BEHOLD...
KERR JARS.
I have never been more excited about canning jars in my entire life, and I've eaten a lot of really delicious food out of Kerr jars.
So, thanks to noticing that, there became this method:
This image shows them all already set up on the testing bench, but they all fit inside a really large cardboard box when I'm driving them from Scott's house to the lab.
Finally!!! I had a way to transport them so that they would stay steady and the concoctions in the vials wouldn't spill all over the place.
So, I know this blog post has been really long and there have been a lot of pictures... I hope that doesn't bother you for whatever reason. Even if it did... um *shrugs*
Anyway, as with every experiment, you figure out what doesn't work and then you start to find things that do and that's always pretty exciting! I hope if you ever pursue some kind of scientific research, you don't get frustrated too quickly. It does require patience, but like I said earlier, sometimes in your most frustrated and helpless moments, you find a solution!
Hope you've enjoyed this journey so far! There will be more to come soon.
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