Just as November disappeared into December, our “preliminary” experiments have become a vital part of our project! As the past two months flew by, Selina and I spent the majority of our time developing a (successful) protocol for the extraction and analysis of DNA from some of last year’s bee gut samples. After amplifying the extracted DNA using PCR (polymerase chain reaction) and visually analyzing the samples using gel electrophoresis, we were left with some varied results and conclusions. First, not every gel we ran was “successful.” Successful in this context refers to the appearance of DNA bands (whether or not DNA from our gut samples could been seen). A variety of potential factors could have influenced the appearance of the results. Among others, some of these might include the presence of PCR inhibitors, the span of time between the initial preparation and the use of the master mix (at later dates), the dilution (or lack thereof) of samples, issues with the DNA amplification, etc. Despite our lack of consistent results, we can conclude that Nosema ceranae is the most prevalent, if not the only strain of the microsporidian present in our county. This was not unexpected, as local beekeepers have not reported signs of Nosema apis (fecal matter on the outside of the hive). Now that our primary testing is complete, it’s time to get creative! Who knows what’s next?
This week, Ingrid and I have been brainstorming ideas on where to take our project now. The one thing we have both been wanting to attempt is to keep honey bees (yes, live bees) in the T.I.M.E. room. So we asked Mrs. William’s and she was all for it! With the help of Mr. Tuckey, Ingrid and I constructed a makeshift “home away from home” for our little friends. I’m sure you’re wondering, “Why on earth would they want or need to keep bees in the T.I.M.E. room?”. Like I said earlier, we are trying to come up with some way to take our project to the next level. One way that researchers have stated that Nosema is transmitted is through fecal matter. Our goal is to collect the feces of the honey bees and test it for the presence of Nosema. The only problem with this idea is that bees have such cleanly habits, that it is difficult to even find traces of any kind of fecal matter within or outside of the hives. That is why we plan on keeping them in the classroom. We (along with the creativity and help of Mr. Tuckey) designed and built our bees a “home” which will prevent them from cleaning up their feces. All it took was a plastic jar with a lid, an Exacto Knife, some mesh wire, a hot glue gun and some cardboard. It didn’t cost us anything and was super easy to make. We cut out a small window and placed the wire on top of it to allow airflow into the “home” and a small hole with a lid in the top which will be how we put the bees into the “home”. The lid will be what we hope the feces is collected in, and inside of the jar, at the very bottom, we glued another piece of wire to allow the feces to fall into the lid. Once the feces falls through the wire and into the lid, the bees will no longer be able to get to the feces which will prevent them from cleaning it up. If this is successful, we will then be able to test the fecal matter for Nosema and allow us a better insight on how and where Nosema is being transmitted.
As the end of October is rapidly approaching, not only are Transylvania County honey bees preparing for winter, but TIME projects are in full swing! Among the monotonous tasks of drafting research plans, organizing stations, and completing a myriad of forms comes the excitement of beginning this year’s research! Honey bees presented countless directions for a project, but ultimately, we decided to further investigate the microsporidian parasite, Nosema. Although Colony Collapse Disorder (CCD) appears to be the most publicized of threats to honey bee health, Nosemosis (the disease caused by Nosema) is perhaps one of the most detrimental conditions a hive can contract. There are two types of Nosema currently plaguing honey bees, Nosema apis and Nosema ceranae. Last year, we were able to identify some kind of Nosema in Transylvania County honey bee hives, but little did we know just how different the two species were and the respective dangers they impose upon honey bee populations. This year, my partner Selina, and I will be using PCR and LAMP techniques to identify and differentiate the two different species and maybe even determine the quantities of each that we have collected. It is our hope that the data we collect this year could help us to later evaluate the most appropriate and effective treatment method(s) for local honey bee hives, so as to ensure healthy and Nosema-free hives for years to come!
This past week, Ingrid and I have been working hard and doing a lot of research to prepare us for our experiments. We have diligently emailed multiple doctors in the field of apiculture (bee keeping) , asking for Nosema spp. DNA and FINALLY we heard back from Dr. Deanna Colby (along with Dr. Kate Aronstein) and Dr. David Tarpy. Dr. Colby is very graciously sending us both N. apis and N. cerenae DNA from Dr. Aronstein’s supply! So while we were waiting, Ingrid and I decided to attempt extracting DNA on our own Nosema samples. We began by filtering our spore samples in order to get rid of all the extra junk we don’t need. Afterwards we centrifuged all samples, removed the supernatant (or the actual DNA), bead beat two of the four samples, and began making gels for our PCR analysis. Next week, we plan on beginning our PCR analysis with both our DNA we have collected and with the DNA Dr. Colby has given us. If all goes as planned, we will know what species of Nosema is present here in Transylvania County!
If you were to look down ¨the bee groups” microscopes these past couple of weeks you would have seen exactly what is pictured. We spent a day preparing the extracted guts for examination and counting by creating a gut and water solution. We then pipetted the 30 milliliter solution into the fantastic disposable hemocytometers we ordered! Then we began the counting which was much more tedious than we expected. Who knew bee guts could hold so much stuff! It was very interesting to look at all of the stuff in the guts that were not nosema spores like small pieces of pollen floating around. We have worked really hard and have now successfully gathered all of our data and are ready to get to a competition!
The past three weeks have been extremely busy for the bee group! Due to the cold weather conditions of the past week, we have decided that our sampling is complete. Upon completing the collection, we have visited six hives total. These hives were scattered all throughout Transylvania County. One of our biggest successes was the Suck-A-Bee, as it worked both efficiently and effectively. and greatly aided in the collection of the bees. The accelerated mite drop test also went very well–it was quite amusing to watch the sugar coated honey bees fly around. Fortunately for the beekeepers in the area, hives have looked very healthy! Overall, the experience was great! We visited some really neat places, met some wonderful people, learned some very interesting information and had a lot of fun doing it! Mite counts are complete and now we intend to begin to the Nosema tests. Aaron has led the group in the gut extraction process and we are all looking forward to getting some results. Hopefully, the spore count will be as low as the mite count and we can conclude that Transylvania County has some very healthy bees!
Lately, we have been removing guts from bees. We are taking out the gut to look for nosema spores located in them. The main section where the spores would be found are in the middle part of the gut where it is darker and larger. We are removing 25 bee guts per hive and mixing them together to take a random sample of guts to look for nosema spores in a hive. The darker the gut, the more nosema the bee is likely to have. We will be testing 10 hives in the process when looking for nosema spores. They must be removed by pulling lightly on the stinger and cutting off the middle gut to keep for testing. As of now, we are in “the middle” of testing our project. We hope to finish soon so we can get results and data on the health of beehives in Transylvania County.
After a week of tediously completing spreadsheets, map questing locations and sending countless emails, Brevard High School’s bee people are ready for some action! The Scorpion vacuum has arrived and the completion of the Suck-A-Bee is in full swing. The Suck-A-Bee is a modification on a hand-held vacuum that we are going to use to collect the samples of bees. After a few minor mishaps and some good laughs, the Suck-A-Bee is finally ready for service. Now, we are focussing on acquiring some last minute bee gear. Fortunately, Transylvania County is home to many friendly beekeepers who have offered to help. With our Suck-A-Bee charged, bee bonnets secure, and pencils poised, we are ready to begin!
Over the past couple of days, Hannah, Ingrid, and Aaron (aka HIA) have been at work completing the “suck-a-bee” in order to collect bees. We have cut out and applied tubing and such in order to collect bees for nosema testing. It seems to work perfectly with no problems. The bees will be sucked up into it and be trapped in a mason jar (Beez in the Trap). Along with it, we have now acquired bee suits to go visit and collect bees inside of hives. We are now prepared to go out into the field, we just need to know where to go. Until we have our hive sites chosen, we plan to do further research and prepare for testing. HIA is ready to proceed to the next level!
On Monday October 13 we presented our groups (Hannah, Ingrid, Aaron) project proposal to the attendees of the Transylvania county bee keepers meeting. It went really well! We gained many more community contacts and we also found many bee keepers willing to let us test their bees in our project! On Tuesday Dr. Wells came to the school and taught us how to properly administer an epi pen in case of anaphylaxis. Now we are going to start contacting the bee keepers and decide when we are going to start heading to their hives and testing their bees!