Brood Analysis Post

        This is just going to be a quick post about frame analysis. First: what is it? Frame analysis is an important step in determining how much the mass of a hive has changed over a period of time. During the hive analysis (where we weigh every part of the hive) we also take pictures of the front and back of each frame (I'll insert one of those pictures below).

This frame has no brood in it, but it has plenty of cells full of honey.

           When we weigh a frame, we cannot tell how much of that weight comes from adult bees, honey, or brood. We conduct the frame photo analysis to separate the brood mass from the mass of other beehive components. Determining the mass of a colony's brood lets us determine how healthy it is by measuring the amount of brood it is producing. Now, how is frame photo analysis done? There are several ways to do it, depending on how the brood is positioned on the frame. 

All the capped cells in this photo contain brood. There is no honey on this frame.

The first step is always to determine the size of the frame in pixels. We do this by selecting the entire frame with the selection tool, and then letting the program (called ImageJ) determine the area (in pixels) of the entire frame. Then, depending on the frame's configuration, we can do one of two things. For the frame above, I simply had to trace all the brood with a "freehand selection" tool and count all the empty cells in the middle with a "multi-point tool" (which counts points). Here's what I ended up with: 

Notice the faint yellow lines tracing the brood. This is how we determine brood area.

However, some frames are a lot harder to handle because the brood is scattered widely across the frame (what we call "shotgun brood") or simply because the shape is impossible to trace. 

A prime example of a frame with "shotgun brood". 

 

This frame's brood was too hard to trace, so I had to count every cell with the counter tool. 

           After finding the area of the brood (in pixels), we input it into Excel. The spreadsheet then calculates the percent of pixels traced from the entire frame. Since all frames are of a standard size, this allows us to estimate how much of a frame's mass is composed of brood. In cases where I am forced to count every cell with the "multi-point" tool, I also input the amount of cells counted into an Excel spreadsheet. However, this spreadsheet takes the average mass of a brood cell and multiplies it by how many cells were counted. Taking all of these steps into consideration, frame photo analysis is probably the most time-consuming part of my data analysis. While most of the data we collected simply require an Excel spreadsheet to be analyzed, this data requires hours (no lie) of painstaking tracing and counting. Even worse, if I accidentally deselect the "freehand selection" tool, I have to start all over again. I can only hope that I'll be finished in time for my presentation. Anyways, that's all for this post. I will probably post again soon about my presentation, or about data analysis if anything new happens.

Mini-Update

       Hi again everyone, this is just going to be a small update post on what I have been doing in the past week. I've mainly been working on continuing to process the data from the experiment, but I've also begun work on my presentation in preparation for May. The brood analysis is going particularly slowly, since a lot of the frames I am processing have what is known as "shotgun brood", where brood cells are distributed widely across the frame. I'll put a picture down below to demonstrate.

The brood cells are the cells of the comb covered with brown caps. In order to assess brood area, I need to trace every cell using a computer program called ImageJ. The "shotgun brood" makes it very difficult to do this.   

        And that's it for this mini-post. I'll try to include another post (as I've said before) showing the kind of data we got and explaining the brood analysis process.

Data Analysis Post

         Hello again everyone, here is the long discussed data analysis post (finally!). In this post, I'll be discussing the kind of data we got from this experiment.
        As I mentioned in previous posts, we observed the temperature, humidity, weight, and carbon dioxide concentration of six hives over about a month. We used iButton sensors for the temperature and humidity data, scales for the mass data, and 5000 ppm NDIR CO2 sensors for the carbon dioxide concentration data. The temperature and relative humidity data from the iButtons were recorded in centigrade and percent humidity, while the CO2 concentration and mass data were recorded in mA, or milliamps. Most electronic scales measure mass by converting the pressure put on the plate by an object into electrical signals. While some electronic scales convert the electrical signals into a mass readout on a display, the scales we used to weigh our hives sent the raw information (in mA) to our data logger where we would download the data later. Likewise, the NDIR CO2 sensors that we used take the spectra of the gasses within the sample space and light detectors at the end of the tube convert that data to mA.
        So, how is all this information relevant? Well, in order to properly analyze our mass and CO2 concentration data, I need a way to convert them from mA to grams (for the mass data) and parts per million (for the CO2 concentration data). To convert the output from the scales into grams, Mr. Meikle usually uses a "weight calibration curve", which is a linear conversion of mA to grams. A similar method is used to convert mA to ppm. However, last time I was at the lab (Monday 4/11) Mr. Meikle couldn't find the weight calibration curve or the conversion from mA to ppm. He hasn't used them in a while but says that his assistant Milagra knows where they are. Long story short, until I get the calibration curves I can't properly analyze my mass or CO2 data.
        We have also run into an issue with our iButton sensors. The iButton sensors are (as I mentioned in a previous post) small temperature and RH (relative humidity) sensors. They look like watch batteries and aren't connected by a wire to a data logger (which has nearly unlimited memory for our purposes). Given their size and the fact that they are not connected to a larger database, the iButtons have short memory spans relative to other data logging equipment. This is something that we neglected to account for. We set our iButtons to record temperature and RH data every 600 seconds (or ten minutes). We "missioned" the sensors (which just means we set them to start recording) on February 29th, 2016. When we checked the data output from the sensors on Monday, we discovered that they had stopped recording on March 9th, 2016. Each sensor had taken about 2000 data points before stopping. When we "missioned" the sensors, we set them to "no rollover", which means that they are set to stop recording once their data limits are reached instead of replacing old data with newly recorded data.
         Between these two dates (February 29th and March 9th), we managed to put in all of our amitraz treatments. However, we only managed to put in one of the three necessary thymol treatments in the thymol hives (each hive requires three treatments) by March 9th. This is a slight setback for our experiment, but we still have 2000 salvageable data points from each iButton sensor. In addition, each thymol treatment is quite powerful so we are at least be able to observe the short-term reactions of the bees to thymol.
        That's all for this post. In my next post I'll be discussing how we determine the mass of brood in our hives, as well as any updates on the data situation. Expect that post sometime this weekend!    

Update Post

          Hello everyone, here's a quick update on what I've been up to for the past few weeks. I was supposed to meet up with Mr. Meikle on Wednesday March 23rd to put in the last treatment of thymol into the hives, but he had a last minute meeting and we were unable to meet up. However, Mr. Meikle put the treatments in later the same day so the experiment was able to continue on schedule.
          On Tuesday March 29th, we counted the Varroa mites on the sticky boards again to compare the figures to those we obtained at the beginning of the experiment. In addition, I began determining brood mass in our hives by analyzing pictures of frames (a technique I will talk about in another post).
          Finally, on Wednesday March 30th, we did our final hive evaluations (which is where we weigh each part of every hive to determine changes in brood mass, honey mass, and adult bee mass). This was also the official end of our experiment. For the remainder of my senior research project, I'll be analyzing data and creating my paper and presentation. However, Mr. Meikle and I are meeting up next week to discuss some of the conclusions I could draw from the data and also some programs that could streamline the data analysis process for me. Expect a post on data analysis soon.

Background Post Numero Uno

          Hello everyone, this in this post I will be discussing some background information on my project. Although I've already discussed some of these details in other posts, I think it would be good to have my entire experimental setup laid out in one post. So, here goes.
          The basic goal of my experiment is to apply two types of miticides (thymol and Apivar) to four hives and observe their effects on honey bee colony homeostasis using CO2, temperature and humidity, and weight sensors. An additional two hives are attached to sensors but will receive no treatment at all (they are our control group). I will insert an image of our work-site down below. This lot contains ten hives, four of which we are not using (they are involved in other experiments). All of the sensors are attached to data loggers which have been put inside lock-boxes for protection from the elements and the bees. I have circled one of the lock-boxes in red in the picture below.

        The orange wires connect the CO2 sensors to the data loggers, and the areas underneath the hives covered in blue tarp are the scales ("weight sensors"), which also connected to the data loggers. The unpainted wooden areas below the hive bodies (which are painted white) contain the bottom boards (which hold the sticky-boards). The temperature and humidity sensors are called iButtons. They are wireless and will have to be taken out in order to have their data read. I will attach an image of an iButton below.

An iButton temperature and humidity sensor.

     Since the iButtons are so small, we placed two in each hive (one at the bottom of frame 4 and one at the top of frame 4). We also put them in small cages so that the bees wouldn't cover the sensors in wax or propolis. Although I don't have an actual image of our sensors' placement within the hive, I will demonstrate their position by using a schematic of a frame.

         The area highlighted in blue is where we placed our nondispersive infrared carbon dioxide sensor. It is nestled between the comb and the top bar of the frame. We decided to place our CO2 sensors here because although CO2 levels in a beehive can exceed 5000 parts per million (and our sensors are only sensitive to CO2 levels below 5000 parts per million), CO2 is heavier than other gasses and usually collects in the bottom hives. For reference, the average concentration of CO2 in our atmosphere today is 400 parts per million.
          The areas highlighted in green and red are where we placed our iButtons (the temperature/humidity sensors). To secure them, we attached their cages to wires and stapled them to the top and bottom of each frame. In addition, we pressed them into the comb so that they wouldn't disturb airflow within the hive as much as they would if they were in between the frames.
         And that's all for this post! My next background post will be about the type of data we will obtain from our sensors and what the data will tell us about the hives. Expect some more update posts soon.  







       IButton EEPROM 256-bit (1). Digital image. Rapidonline.com. Rapid Electronics, n.d. Web. <http://www.rapidonline.com/ibutton>. 

      Cushman, David A. Dummy Frame. Digital image. Dave-cushman.net. David A. Cushman, n.d. Web. <http://www.dave-cushman.net
             /bee/dummyframe.html>. 

Pardon Me if I'm too Blab-bee

          Hi everybuzzy, here is another update post about what I did during the week. I went in to the CHRBC twice this week- once on Monday and once on Wednesday. On Monday, Mr. Meikle and I did hive evaluations on all the hives involved in our experiment. In hive evaluations, hives are taken apart piece by piece, and each piece is weighed individually on a scale. Pictures are also taken of each frame so that the amount of brood in each frame can be counted and an estimation can be made of their weight. Since the hives are already mounted on scales, the weight of all the components can be subtracted from the hive's total weight to obtain the mass of all the adult bees in the colony. The parts we weighed were: the hive lid, the box, the entrance reducer (which controls the amount of bees that can enter and exit the hive), the tarpalet (a small tarp that covers the hive scale), the strap (which keeps the lid fastened when hives are being moved), the frames, the riser (which raises the lid about an inch to make room for sensors and/or pesticide treatments) and the bottom board (a small drawer with a sticky-board fixed on top that catches all the detritus that falls from a hive).
         After weighing the components of all six hives, we removed the sticky-board from the bottom boards and brought them back to the lab. A sticky-board is a thick piece of paper covered with Vaseline that catches everything that falls from the frames of a Langstroth beehive. The purpose of putting the sticky-boards in place is to catch Varroa destructor mites that are removed from recently vacated brood cells by bees or fall off their hosts. Beekeepers and researchers can obtain two key pieces of information from sticky-boards- they can see how heavily infested with mites a beehive is by counting the mites on its board, and they can determine how many mites are falling per day by dividing the total number of mites by how many days a board has been in place. In short, we counted the amount of Varroa on our hives' sticky-boards once we got back to the lab. These hive evaluations are crucial to our experiment because we need to know where the hives are at before we apply the treatments so we know how much the treatments affected the hives.
        On Wednesday, we applied the treatments (Apivar and Apiguard, whose active ingredients are Amitraz and thymol). I'll include a picture of what they look like. Apiguard is a gel that is placed onto cards which are put on top of a beehive's frames, and Apivar comes spread onto plastic strips that are inserted in between the frames.

Apivar strips.

Apiguard gel. There is about 25g of gel on the hive tool (yes, that's what it's called), which is the recommended dosage for an average beehive.

            That's it for this week's update- I'll try to get in some "background" posts soon (but I'd like to get some pictures on-site first).

Packshot_apivar. Digital image. Apivar ®. Veto-pharma, n.d. Web. <www.veto-pharma.com/products/varroa-control   /apivar/>. 
Oliver, Randy. Apiguard Gel, Bulk Pack. Digital image. Scientificbeekeeping.com. © Randy Oliver, n.d. Web. <http://scientificbeekeeping.com/ipm-7-the-arsenal-natural-treatments-part-2/>.

Update

          Hello again everyone, this has been a very buzzy week for me (which I'm eager to tell you about!). But first I'd like to clarify some things about my future blog posts. From here on out, I am going to start doing two different types of blog posts. The first kind will be updates on my SRP, and the second kind will be general information about bees and background on my project. However, today's post is simply an update.
           Before I talk about what I did this week at the lab, I want to discuss the purpose and the methods of the experiment that we intend to carry out. Using CO2 sensors, temperature and humidity sensors, and weight sensors, we intend to determine the effects of different miticides on bee colony homeostasis. We will be testing two miticides: thymol (a substance extracted from thyme) and Apivar, a miticide commonly used by beekeepers because it kills Varroa mites without leaving significant residues in honey or wax. Thymol is often applied as a gel, while Apivar comes embedded in cardboard or plastic strips.  I have six hives at my disposal for this experiment. Two of the hives will be given the Thymol treatment, two will be given Apivar, and two will be given nothing at all (they will act as our control group).
          Now, about what I did this week. In preparation for Wednesday (when we installed our CO2 and  temperature and humidity sensors in our hives) I went to the lab on Monday to familiarize myself with the CO2 sensors. This was also Dr. Meikle's first time working extensively with these CO2 sensors, so he had a sensors specialist talk to us about how they work. We are using Nondispersive Infared (NDIR) CO2 sensors, which are spectroscopic sensors that detect CO2 in a gaseous environment by its characteristic absorption. The key components of NDIR CO2 sensors are an infrared source, a light tube, an interference filter, and an infrared detector. In the sensors that we are using, gas diffuses into the light tube through a membrane and the electronics determine how much CO2 is present by measuring how much absorption of CO2's characteristic wavelength of light there is. We have at our disposal four 5000ppm CO2 sensors and two 2000ppm sensors. After learning about how these sensors work, we set them up in the lab and tested them. Here is a picture of what we did: 

This is our CO2 sensor setup. The gray tube is a 5000ppm CO2 sensor.

          On Wednesday, we got into our bee suits and installed the CO2 and temperature and humidity sensors (I will go into more extensive detail on our setup in my next post). We also installed a gridded board in the bottom of the hive that will catch falling Varroa mites. By counting the number of mites in each square and dividing by how many days the board has been in place, we can determine mite fall per day and, as a result, mite concentration. Expect more updates soon- I will be going to the CHRBC (Carl Hayden Bee Research Center) twice next week.

This 'Mite' be a Problem

          In 2006, seemingly healthy bees began simply abandoning their hives en masse, never to return. This phenomenon, called colony collapse disorder (or CCD), endangers the livelihood of the beekeeping industry and agriculture today. Many researchers point to crop pesticides, global warming, habitat loss, parasites, disease, and even cell phone signals as potential causes of CCD. Although the cause of CCD remains a mystery, one thing is for certain: parasitic mites are a very real problem for bees and beekeepers around the world. Although mites like Varroa destructor and tracheal mites often don't kill bees directly, they often disable them and render them unable to perform the daily tasks needed to keep a hive healthy.

The Varroa destructor mite.

        Many beekeepers use pesticides (or miticides as they are sometimes called) such as Apistan, Hopguard and thymol in order to reduce or eliminate mite infestations. However, the amount of pesticide that will kill mites isn't much lower than that which can hurt or kill bees. This poses a problem for beekeepers, who are unsure whether they are hurting or helping their hives by applying miticides to them. 
        The objective of my project is to evaluate the consequences of pesticide exposure on colony growth and brood production, forager activity, and colony homeostasis to determine if pesticides are indeed having a detrimental effect on honeybees’ well-being. I intend to monitor the vital signs of two colonies exposed to miticides (with two given blank miticide strips and another two with nothing at all) using temperature, humidity, weight, and CO2 sensors, with the help of William Meikle at the Carl Hayden Bee Research Laboratory in Tucson. I am very excited for my project- if all goes well I will have some neat results. I'm going in on Monday to familiarize myself with the CO2 sensors, and we will be installing the sensors on Wednesday. I will post updates as they happen, but I 'hive' to go now; we'll talk again soon.