Honey Bee Micro-Nutrition and Pollen with Dr. Ramesh Sagili (308)

308 - Honey Bee Micro-Nutrition and Pollen with Dr. Ramesh Sagili

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Jeff Ott: Welcome to Beekeeping Today Podcast presented by Betterbee, your source for beekeeping news, information, and entertainment. I'm Jeff Ott.

Becky Masterman: I'm Becky Masterman.

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Jeff: Hey, a quick shout-out to Betterbee and all of our sponsors who support allows us to bring you this podcast each week without resorting to a fee-based subscription. We don't want that and we know you don't either. Be sure to check out all of our content on the website. There, you can read up on all of our guests, read our blog on the various aspects and observations about beekeeping search for, download, and listen to over 300 past episodes, read episodes transcripts, leave comments and feedback on each episode, and check on podcast specials from our sponsors. You can find it all at www.beekeepingtoday.com. Becky, you know what I want for Christmas?

Becky: What's on your list, Jeff?

Jeff: Listener openers.

Becky: Oh my gosh, what a great new year would be if every single week we had a listener opener.

Jeff: Yes. I think that would be good, and I think our listeners would enjoy that little stocking stuffer as well.

Becky: I totally agree. I agree. Let's make that happen. We will wish it for Christmas. How's that?

Jeff: It's out there. It's out there now.

Becky: Very good.

Jeff: Becky, how are your bees doing there? Have you been out to check on them?

Becky: I wish I knew how they were.

Jeff: Little stockings hung on the front of the hives.

Becky: No. I've resisted the urge to go and do any peeking and checking. I'm without any knowledge right now, but I'm hopeful and I know I certainly did my best to prep them for winter. I told them, you've got these girls and I'll be back when the weather's appropriate.

Jeff: They said, please don't come knocking.

Becky: Right. They're like, you can leave us alone too. We'll see you when everything warms up a little bit.

Jeff: Good. This time of year is always very difficult for beekeepers and hoping that the bees get through the winter okay, especially those in the northern tier states. Those in the southern tier states, the bees are probably still moving about. Here, we'll have a cold snap and we won't see a bee for several weeks. Then if January holds like many Januarys, we'll have warm spells and the hazelnut will start bloom, and the pollen pods will come out. We'll see bees working pollen, the hazelnut in January. It's always like, oh, it's nice.

Becky: It's usually early March. That's our first pollen coming into the colony, so we have to wait a little bit more. They do start rearing brood in January. We know that from previous data. Hopefully, they've got that pollen stored in the comb that they're in so they can use that protein to raise their new bees. It's December, so there's probably not a lot of brood in there right now.

Jeff: I'll tell you though, I typically see several of the colonies based on where I have a sensor located, that they keep that area pretty warm throughout the winter and some of the colonies. I always figured it's either all the colonies or select colonies have a little pocket of brood going, but it's fun to see that.

Becky: Definitely, I've caught them raising brood in December for sure. It can definitely happen, and I think it just depends upon the colony genetics and what the conditions are. I think some of the answers, we don't have as far as why they will continuously raise brood rather than shut it off during the winter dearth. It's a mystery.

Jeff: Something yet to be decided. On today's guest, I'm looking forward to talking to Dr. Ramesh Sagili of Oregon State University. We had the pleasure of meeting him in person at the Washington State Beekeepers Association meeting back in October. He and his team are providing some great information to the nation's beekeepers.

Becky: So impressive. It's a great, great group. You know what really impressed me about his graduate students and his undergrad team, they were just so happy at that meeting. They just seemed really grateful to be in a room full of beekeepers and talking about the research they're doing, and it's just always so impressive to see that kind of a team get out there and do their best to support bee research.

Jeff: We're going to be talking to Dr. Sagili right after this quick word from our sponsors.

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Jeff: Hey, thanks a lot to our sponsors. Hey, everybody, sitting across our great big Beekeeping Today Podcasttable is Dr. Ramesh Sagili from Oregon State University right down the road for me in Oregon I5. How you doing, Ramesh?

Dr. Ramesh Sagili: I'm doing well, Jeff. Hi, Becky.

Becky: Hello. Nice to see you again.

Jeff: We had just seen Dr. Sagili at the Washington State Beekeepers Association meeting back in October and we're really happy to have him here today. Ramesh, why don't you tell us a little bit about yourself, your background, and how you got interested in bees?

Ramesh: It's a long story, but I'll keep it short. I grew up in India and then I earned my bachelor's and a master's degree in agriculture. Then I came to the United States around 2002 to pursue a PhD in entomology, focused on honeybee research. Mostly I was doing a brood pheromone research, looking at foraging behavior of honeybees. I stuck around another year and a half for a short postdoctoral position at Texas A&M as well. Then I was fortunate to get this assistant professor position at Oregon State University around 2009. I've been here about 15 years now.

I'm in the full rank of professor here at OSU. My program focuses on three primary areas, honeybee health, which is very broad. Anything from Varroa mites to Nosema to viruses. Honeybee nutrition has been a serious focus for the last 10 years, I would say. Then pollination, because my position is supposed to work closely with the stakeholders, being our commercial beekeepers and farmers. I closely work with some pollination projects as well. I also say sometimes CCD or colony collapse disorder is a blessing in disguise for the bee industry.

As well as some people like me who were hired after CCD around 2009 because my predecessor had retired, Dr. Mike Burgett, around 2002, and there was no bee position for almost seven years. There was a lot of push from the industry, both the pollination industry that needs bees for pollination. Of course, beekeepers, as you know, it's a small sector, they don't have that much of power like farmers, like cotton farmer or soybean. I think this is all great. CCD, I know it was unfortunate, but as I said, it's a blessing in disguise. There were many positions that were awakened at different universities, were hired back, including the one I'm at currently.

Jeff: What made you interested in honeybees?

Ramesh: People are surprised when you say you are doing honeybee research. They say, oh, can you just focus on one insect and do research on honeybees? My story is, as I said, I have a background in agriculture degrees, but my background goes to high school when really at that point, I don't think I decided that I would study honeybees. My grandparents were farmers and so they were growing or cultivating hybrid sunflower. In those days, hybrid sunflowers, as you know, you need a lot of cross-pollination. For some reason, they didn't have enough honeybees and even other native bees around.

Could be because of indiscriminate use of pesticides or whatever. the reason was. They would hire these people, and they would wear gloves, and then they would touch each flower head and go to the next one. They're doing the job of a bee, technically. I would say, wow. 10 years old, I didn't really understand. I said, wow, why do we do this? They said, because we don't have enough bees. That was my first impression, how critical bees are for pollination, and sometimes we take them for granted.

In some countries, that's a real situation where you don't have enough bees for many reasons. Again, I didn't decide to study any bees at that time, but when I took an apiculture class as a part of my Bachelor of Science in Agriculture, that's what I thought probably-- Entomology was always my passion among all those different subjects they teach. Again, bachelor of science is a little different in India than here. You can't focus on just one entomology or one plant breeding, or something. They make you jack of all trades, so you learn everything from--

I took a lot of entomology classes, plant breeding, soil and crop pathology. Ultimately, in those countries, the goal is that you serve the farmer after you become an agricultural graduate if they come up with a problem. Farmers would bring a rice or the paddy total plant and say, I have this root worm or whatever, how do I deal with this? You need to have this extensive knowledge of entomology. Again, coming back to entomology, that was my passion. I thought I would do higher studies in entomology, and I thought bees would be the real good ones to do so.

Jeff: Bringing a lifetime of interest and exposure to what you do today is really fun.

Ramesh: Absolutely, yes.

Becky: Ramesh, did your college class take you to an apiary or did you work bees?

Ramesh: Not extensively. It was just an exposure just for the class time. You just go for that one hour class and just do some manipulations initially, but no, not an extensive beekeeping.

Becky: That's how I got started at beekeeping, that one visit to the apiary in my college entomology class. We share that in common.

Jeff: You run the bee lab at Oregon State University. It's a really, really strong program, especially after talking to your students in October, how excited they were about being part of your program and the bee research that was going on. You mentioned that one of your areas is nutrition. That's an area that is starting to get a little bit more focus these days. What aspect of nutrition really inspires your curiosity?

Ramesh: Great question. When I started looking into the subject of bee nutrition about 10 years ago, I would say, seriously, I would not find enough literature on micronutrients. We talk about macronutrients and micronutrients for bees. Macronutrients, for your audience, would be your carbohydrates. Your honey or nectar is your carbohydrate source. Then you have pollen, which is their only protein source. We have adequate knowledge in that front. Then there are micronutrients that are present in pollen and nectar, which are really critical for the growth of an insect, and in our case being honeybees, which surprisingly, we have barely scratched the surface.

Sometimes I joke about this because at Texas A&M when I was a student in a PhD program, there was another scientist who was studying grasshoppers, and they were studying grasshopper nutrition. I was surprised when I came into bees here that we know more about grasshopper nutrition when compared to honeybees, especially in terms of micronutrients. There are reasons. It's not like honeybee has been studied very well, at least in the last 200 years in the modern world. We know a lot about honeybees, and it's a very important beneficial insect.

Grasshoppers, it's easy to study, because I tell about this complexity in nutrition with honeybees, is there is a colony-level nutrition, there is an adult, then there is a larva. Larva is eating something very different when compared to what your adults are eating. That complexity makes it hard to study very focusedly on some aspects. For grasshopper example, you have the adult that's feeding on a plant, some specific species, and then the nymphs that come out also feed on the same thing. The grasshopper is not feeding a very specialized diet like your honeybees are having those hypopharyngeal glands and mandibular glands in their heads that are producing that brood food, which is critical for larval survival.

Again, that's the reason why I think we have not studied that well honeybee nutrition because it's also very complex to study as well in a more controlled setup. Coming back to your questions, I'm glad that there are many people are interested in-- When I started doing 10 years ago, there were a couple of other labs that were interested in bee nutrition, but now I see there is a growing interest in this area.

There are more labs involved, and I'm glad you guys are doing this podcast on habitat as well, which is really critical to understand because honeybee nutrition is so important. I know we have bigger problems, Varroa, other things too. If we deal with nutrition, which probably we can do as beekeepers or farmers, or conservation groups, that would be really cool so that at least our bees can take a little more stressors. I'm not saying we should have more stressors, but I'm just saying at least whatever we have can be at least mitigated to some degree if we have better nutrition.

Jeff: Other guests have said a well-fed bee is a healthier bee and can, like you said, withstand the stressors a lot better. If you go into the winter with bees that are undernourished, they're certainly going to have a hard time through the winter months and being ready for spring.

Ramesh: Yes, because there is this vitellogenin. For the audience, it's a glycolipoprotein, vitellogenin, very critical for longevity of bees, and pollen, which is the important source of sterols and everything. That's when you don't have pollen, then you don't have sterols, and then you don't have vitellogenin as well. That's really critical. Your bees have to be really well fed. At least they should be in a better nutritional state by September or October so that that's when they raise those winter bees. We use this scientific jargon, diutinus bees, which is a Latin word for long-lived bees.

Those bees are usually raised in late fall. If your nutrition is compromised at that time, then those bees probably-- I get this question all the time. Hobby beekeepers will call and say, my hive was strong, beautiful, 25 frames or whatever of bees in October, then in December, I went and they all died. It could be because of Varroa for sure as well, but even nutrition as well. Those bees were not raised on an optimal diet.

When you don't have that optimal diet, then probably those bees look good physically. I may look physically okay if I'm not eating right, but I will get sick soon. It's the same thing with honeybees as well. If you are not eating right when you are being produced, so that's probably is compromised immunity as well as the longevity of life as well. That's how you see those bees all die in December or January, and you don't have enough bees left.

Becky: Ramesh, I've just got to ask, what does it look like if they die because of nutrition versus die because of Varroa at that time of year?

Ramesh: Great question. I wish I had a good answer for you.

Becky: I didn't think there was one, but I decided to ask.

Ramesh: That's great. What we have been trying to do-- Hannah, one of my technicians here in the lab, so we are trying to come up with a trehalose assay. Trehalose is an insect sugar. For the audience, if you're confused what trehalose means, it's an insect sugar. It's a hemolymph. It's a blood sugar for insect. We measure our blood sugar with anything like your meters that you have at home, those who are diabetic or whatever. We can measure the trehalose in the hemolymph, which is the blood of the honeybee.

We are trying to come up with a good protocol where we can see if there is less of this, then probably these bees are starving. Maybe we can also measure protein levels in the hemolymph as well, not just the sugars as well. That could tell if the bees are dying because of starvation, versus they have died because of something else. Again, these are still under works, but Becky, great question. I wish we could give a simple answer to our audience that they can just take 10 dead bees out and say these died because of starvation versus mites.

Becky: It sounds like you're thinking of some kind of a fall assay where you can actually check for mites and also check for nutrition.

Ramesh: Yes.

Becky: That's exciting.

Jeff: That'd be really good. If you're going to sacrifice 300 bees for a mite wash or something like that. Also get, if you could, in a nice world, it would be nice to be able to also get nutritional information, status from those.

Ramesh: In general, people have this idea that, ah, the tongues have stuck out. Sticking tongues is not really a good analysis. Sometimes, I know, you'll see bees stuck their heads in. Yes, that's a typical starvation because they didn't have enough nectar or honey during winter. Then some people will just make a assumption that-- even pesticides, people think the tongue protruding out and dying is an indication of a pesticide kill or starvation. It's not that black and white. Some cases probably is. Yes, you need a better, more physiological test for those rather than just looking at the symptoms.

Becky: It's kind of what you said about grasshoppers, too, but if you compare what we know about honeybee nutrition to what other livestock industries know about their animals, we're still not quite there. There's a lot of work we have to do, right? You have to do it, actually. You've taken the challenge on.

Ramesh: Not just me. We have other colleagues at other universities as well. That's what I appreciate is again, one of my postdocs, you probably know Priya. She's been doing some of the physiological work as well with nutrition at Mississippi State University. There are other colleagues in Tucson and even Penn State and other locations where they're studying nutrition. I'm really happy that in the next 10 years we'll have more knowledge that would really benefit beekeepers in terms of bee nutrition.

Becky: I remember a commercial beekeeper said to me once. He said, "Did you see my bees? Did you see how fat they are? My fat bees are really healthy bees." I thought, oh my gosh, is that something? Bee size can definitely be, depending upon the cell size, but it could also be nutritional. He might have been onto something?

Ramesh: Yes.

Jeff: Let's take this quick break. We'll hear from our sponsors and we'll be right back with Dr. Ramesh Sagili, Oregon State University. We're talking about nutrition and we're going to get onto some other subjects.

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Becky: Welcome back, everybody. Ramesh, you mentioned Priya, and Priya is one of your collaborators on the pollen national database. Can you talk about that a little bit?

Ramesh: Yes. Priya was a postdoc in my lab almost four years, five years. I lost track now. She's a assistant professor in Mississippi now. At that time we started this work. I think we submitted a USDA grant in 2020 when she was here. It's an exciting project because what happened, this pollen nutritional composition database, and it really dovetails with your podcast of improving habitat for bees is, you probably have heard from a lot of other researchers and others. They have some recommendations what to plant for bees maybe in summer, spring, winter.

Not winter. Late fall or whatever for bees, but most of these recommendations are coming from attractiveness, if you give a thought. Some people will ask you, probably both of you, Jeff and Becky have gotten questions and even the audience probably has gotten questions about, give me top 5 plants I can put in my backyard or top 10 plants I can put as a farmer somewhere to help bees. How are these coming? They're coming from some prior experience, from someone-- We consider that as an attractiveness. Someone has looked, oh, I have this lavender, there are hundreds of bees coming onto this.

Or, I have bachelor's button or borage or Phacelia or Sainfoin, or whatever. As far as my knowledge, it's coming from attraction, but attractiveness, so think about this. I always give this lame example of French fries. You give me French fries and salad next to each other. What would you guys eat? You'll probably try, like me, some French fries first and then start eating salad. French fries are so attractive. That's the same. I know it's not a great analogy, but think about that. You don't have an option there and just will go for the French fries. If something, is attractive doesn't always equate to better nutrition.

That's the argument I'm going to make is, I'm not saying what we have this time, all this beautiful databases. USDA has a plant database. You've probably heard of that. They have list of plants. Similarly Portland, that there is a [unintelligible 00:24:01] society which is an invertebrate conservation group. They have a bunch of plants for different regions that you can plant. We wanted to go a step further from there and say, these plant lists are there, but wouldn't it be nice to put all these pollens together? Collect those pollens, look at the nutritional composition, just for pollen.

I'm not talking about nectar yet. That's another bigger endeavor in the future. Just pollen, you collect that and look for all the micronutrients that we are worried about. Phytosterols, look at crude protein. That's not a micronutrient, it's a macronutrient. Then look for amino acids. Look for omega-3 and 6 fatty acids, which are really important as well. All these different things that you can look for in the pollen. Then wouldn't it be nice if I put another column next to it? By the way, the USDA NRCS and all these plant database people are really excited.

They're waiting for our information to be available so that they can put it next to their list of plants. A column where it says, you have Phacelia, crude protein, amino acids, lipids, omega-3, 6 fatty acids, everything that we can do comprehensive. There really so many that we'll miss, but we'll try to do as best we can. That being a huge endeavor. The first step is to collect pollen. Collecting pollen is a painful task if you have tried before. We are not collecting from bees. Only in case where it's hard to collect hand collection we go with bees. We have citizen scientists across the nation.

Probably we have recruited at least close to 100 I guess at this point. They are collecting pollen for us. First six months or a year, one of the graduate students, it took time to really design a vacuum. We have a vacuum with three partitions. Pollen comes in different sizes, could be anywhere from 200-plus microns to 30-micron size. You want to collect all those different sizes. If you imagine this is a 2.0 for the audience. I'm making some hand gestures here to back in. If you are imagining the tube in the front for a vacuum, so we make three partitions.

Each partition has a different mesh on it. When you go and put onto the flower and start the vacuum, the pollen will be sucked into that. If it's a smaller size, it'll go deposit on the last one which is about 30 microns or so. The bigger ones like 200-micron size, they'll stick to the front of that mesh. Then you take the brush and it's a very painful process. You have to do thousands of flowers sometimes to collect, but someone has to do that. That's a hard work. That's the reason it's been three years since we got that. The grant is done as well, the money we already spent, but still we are trying to analyze all that part because it took so long to collect 100 species.

Currently we have about 100 species that we have collected. These include anything from natives to crop pollens, from blueberries to cherries to apple, whatever you can think of. It would be a very robust data set eventually once it's available. That might be helpful for stakeholders. All these conservation groups, beekeepers, farmers, whoever wants to plant something. Now they can choose those plants based on a more scientific way rather than just randomly selecting some that might be suitable for their area or whatever. I think that's why we are really excited about this project.

Becky: Is your assay work separate from the nutritional database work or are you informing it?

Ramesh: You mean the assay for all the nutritional companies?

Becky: Right.

Ramesh: Collection is one part. Then we have a mass spectrometry lab downstairs here. I'm on the fourth floor here in this building where I'm talking from. Priya's lab has helped to prep the samples, and then they come back to OSU. Then the mass spec lab downstairs. We have given them all the instructions, what we are looking for. They have made standards for all the phytosterols and other things that we are looking for. That part happens here. We are halfway probably on the nutritional analysis, but to get everything done, maybe another six months and we should have 100-plant list available. Then probably we'll ask them to put on some. We'll have our own website too, but I think USDA NRCS is excited about having this composition data for their website as well.

Jeff: Using that information that you develop and you publish, how would beekeepers use that then? Would that be the beekeepers or would that be the people who are designing habitats? How do you propose that be used?

Ramesh: It's for all. All the audience that you mentioned. As I said before, conservation groups because they are more interested in large acreages to plant. Even Pam has given a lot of seed to plant in California almonds and other places. That's a nice effort. Maybe once we provide all this information, even those type of agencies that are interested in improving habitat can use this data. We have a plant list from different agencies. We are providing additional information that might be really useful to make a more scientific choice rather than just randomly selecting something.

Coming back to your question, beekeepers, if they want to plant something in their small acreage, 5 acres or whatever they have for their bees, if they're commercial beekeepers, then they can choose. They can use the same list and look for-- suppose, just for example, I'll make here something. You have 10 plants I'm looking at here, and there is something like buckwheat or maybe here as alfalfa or some of these other natives, or Russian sage or something, Sainfoin. Then you look and say, these are all suitable for my region. Then you look for the nutritional composition.

Right now you have additional thing to look for. Then you look at crude protein, 20% phytosterols, this many nanograms, whatever we gave, the milligrams per gram. Based on that they can say, oh, looks like this is low and then they go to the next crop. They can make a combination of those. Even if you want to plant only five, then you can choose those top five ones that have the optimal crude protein, optimal sterols, and those important things that we are looking for.

Jeff: I could see where it also come in handy where if you had a choice of bee yards, you could say, oh, there's a lot of flowers, but you look at the chart and it's not that nutritional where this yard would be much more nutritious for my bees.

Ramesh: I'll add one more thing, Jeff, for this. Probably Becky has read a lot of nutrition as well. We talk about protein-lipid ratios as well. Different times of the year. Logically thinking, bees are going into wintering phase. Close to September or October. We would imagine that they will feed on proteins naturally available if they are able to detect, which I haven't seen any studies that honey bees can detect protein concentrations and lipids, but some bumblebee work has been done that they claim that bumblebees can detect somehow the protein content and other things. Again, I want to still see more work on it, but I'm not really very confident at this time.

Those are nice studies that were done. What I'm saying is, so the whole concept is you probably need more lipids. Because bee are storing that vitellogenin and more fat bodies in their bodies to survive the winter. Logically thinking, you should expect that the protein or the pollens that they bring in and fall or late fall should have high lipid content and probably still decent amount of protein. Those things can be useful as well. Once that science is available, then maybe this dataset can be used to see, this plant species has high lipids, so maybe let's plant this for bees to be available in fall or late fall so that the bees would benefit. I can see a lot of different directions this can go.

Becky: Ramesh, how important would a nectar nutritional database be?

Ramesh: Oh, that's, again, that's huge. As I said, we are still already drowning in pollen, so I don't know when.

Becky: I don't want to rush you.

Ramesh: I don't know when the nectar will start, but nectar has a lot of phytochemicals as you know. They're really important. I think one of the labs, I think May Berenbaum's group, they published something on phytochemicals, how they're important like the p-couramic acid and those things. Nectar is really important as well. We are not discounting the-- pollen, why we chose is it's critical for colony growth. That directly is impacting your brood rearing.

That's why we thought let's start with pollen, nectar will do deal at a different time. Maybe someone else is already doing, I don't know, but someone has to do. I'm not saying we have to do that. Someone else can pick that up as well because nectar also has a lot of important nutrients, not just your carbohydrate. Some phytochemicals might be really critical that are present in nectar too.

Jeff: Related to nutrition is an early paper you did, and we talked about it briefly before we started recording, but the early paper you did on the larval nutritional status versus the genetic relatedness the nurse bee and breeding or raising bees and maybe the queen rearing. Can you talk a little bit about that? Because that's something that's really not discussed a whole lot.

Ramesh: I'm glad you dug into my previous work, which I had totally forgotten about this last paper until you mentioned.

Becky: It was a serious vetting process to be a guest on this podcast, Ramesh.

[laughter]

Ramesh: Thanks, Becky. This was my last project, my PhD work and I didn't find my enough time after I came to Oregon State University because we had so many other projects going on. It was still in my ladder, but not-- We published that work. It's really intriguing. You probably heard of kin selection. For the audience, to give them some context here. We are talking about emergency queen rearing here. As some of you probably understand, emergency queen rearing is when there is a disaster, something happens to the queens all of a sudden.

It dies because of a virus or some other disease, or maybe as a beekeeper you are not careful and you kill the queen. Then probably the bees will realize. They will know there is no queen pheromone. Within several hours, they'll figure out that they have to raise a new queen and they start making, because that's not an ideal scenario for them. They really want when they do make queen cells for swarming time, they're very meticulous in selecting if there is an emergency queen rearing or whatever. When they are suddenly asked to choose, they will choose till some--

As you all know, they are totipotent. The larva can be raised until they're three days old. After that, probably you'll not get a good queen. This study, what we were trying to really prove is, because previous studies in the '80s and probably some in '90s, they had this hypothesis where they thought it's kin selection. Again, to give you some background, if your queen met with about a dozen drones, so there are a dozen patrilines in the hive. If I am a larva that is related to 1,000 other sisters I have, so we all may have some bias to select-- if we are the bees from one group, same dad, then we all have this bias towards some larva that are related to us.

That means the queen has laid, our mom has laid eggs, and there are larva and there are some related to us. We may have this urge to pick larva that are related to us and make a queen out of this because now it's an emergency scenario. Yet all many of those bees that are inclined to do this have a choice to make. What we wanted to dis screw is, or at least to prove that kin selection is not-- even those publications that were on kin selection, they had said there is a very small chance, they were not very conclusive studies. We thought maybe nutrition plays a role because nutrition is so critical for bees, so we thought let's starve some larva.

I know you can't starve larva right there. You are compromising the design if you're starving. With all our previous work on larval pheromone, that was my entire PhD was on larval pheromone, we knew what age to starve and then without impacting their fitness. Fitness means-- you probably understand. We chose the larva and only start for four hours because our previous study it has shown if four hours, the size doesn't change. We take a frame, again, I know I apologize you can't visualize what gestures I'm making here to Jeff and Becky. You have a frame, just visualize a frame of full of larva.

What we do is we insert a mesh, so goes into a square mesh on this side or right side and left side. You're having an equal number of larva that are pushed in. One mesh will have a bigger mesh where the bees can go and feed the larva and on the left side will be a mesh, which is a smaller dimension where the bees can't reach the larva to feed. Those will be start for four hours. After that you remove those. It's very neatly done. Jeff, I'll send you the link for your audience. They can read by themselves, but I don't have time to really explain everything well. Actually we cut those frames into half as well.

Some of them had the nails on the top, so we would swap them into different hives that are queen-less so that everyone gets one set of larva that is starved and one set without starvation. Then we allowed those foster colonies that were queen-less to raise their own to see which larva do they pick to make a queen. It's not like they totally ignore the four-hour starved larva. They have same size. We looked at several of the parameters. Again, read the paper if you're interested. It was a neatly done study where we didn't compromise anything except that there might be some compromisation in the larva pheromone.

We think the larva that were starred for four hours, they were somehow releasing the pheromone, which could tell the bees that these are inferior. Whereas the well-fed ones probably had the full pheromone blend, they were indicating that we are the fit ones. What we saw, again, to summarize, we saw less number of larva that were selected from the starved group to become emergency queens. There were more significantly higher in the non-starved group. That's how we figured out that nutrition is involved and bees can recognize a larva based on their nutritional status that we should not pick this one and we should pick this one to become a future queen.

Becky: Very interesting and elegant.

Jeff: Goes to show how, not only in overwintering, which was our example when we started the discussion on nutrition, but it's also if anybody's raising queens to make sure that all their queens are well fed, well nutritioned.

Ramesh: That's another lesson. I know it's not something like rocket science for queen breeders. They already know much of this, but I think it's just another reassurance that nutrition is really important for those bees and those larva that are being picked for grafting to make a new call.

Becky: You also have interesting information about European fall brood and nutrition that you've been working on. Can you maybe share that in our last minutes together?

Ramesh: Sure. This is another exciting project. It's a multidisciplinary multi-state. I lead that project, but my colleague, Dr. Andony Melathopoulos, next door here, he's a big part of the grant as well. We have been getting these reports, and like Jeff, you probably heard you are in some locations where blueberry, I know Minnesota doesn't have, but Michigan next door has a lot of blueberries. That's been a big problem for many commercial beekeepers. They have been complaining for the last, at least six or seven years that the bees don't look good and they get a lot of EFB, European foulbrood disease, it's a bacterial disease, for your audience.

I know some Michigans, some other groups are also leading this effort to some degree, but I haven't seen any comprehensive effort to look at a lot of different factors that we think. This is a longitudinal study. We are following about 1,500-plus colonies from 4 different states, including Washington, Oregon, California, and Mississippi. We are looking for triggers that are responsible for, or what are the triggers for EFB when they're in blueberries. We don't think it's just blueberry has something magical that is creating the problem. I think there are several factors.

It creates a perfect storm when they go to blueberry pollination. We are tackling a lot of different factors, including nutrition. I'll stick to nutrition because of the time here. Nutrition has been hypothesized as a big factor in the past as well. Again, there is no real conclusive evidence that has been shown. We are trying to gather that. What we do, again, I'll make it very brief for you. It's a longitudinal study. It'll take a long time to explain. We are taking a lot of different things. We score the hive when they are in blueberry. The study starts in California. We have all the 1,500 per sides, they have a tag.

They say OSU1, beekeeper, name, date, everything. We longitudinally follow when they come to blueberries, they go to Oregon, then Washington. Brandon is from Washington. His group is following those sets of his hives. When I say his means his beekeepers. There are four beekeepers from each state that are a part of this project. We look for everything. We score EFB with a 1, 2, 3, depending on how many larvae are infected. Then we also collect nurse bees from brood area to correlate their nutritional status. Again, I just got some basic information, our Oregon conference is this weekend.

On Saturday I'm going to present some of the similar slides, what you Jeff, and Becky saw there but with more data now I have already enough data that I can show them. What we are seeing is that the bees, that there is some correlation, at least now with some preliminary data we are showing is. We wanted to see if a score of EFB 3 means high. Then I'm also looking at vitellogenin levels at that time. If the vitellogenin levels are changing, suppose for 3, we expect the levels to be low. That's why the disease is increasing here. We are seeing some very cool patterns already with the nutrition part, but again, it's very preliminary.

I don't want your audience to take it very seriously at this time. Maybe next time I speak I can give you more robust information. Looks like nutrition also has a major role to play as we discussed in the beginning of the podcast, that nutrition can help you mitigate a lot of other stressors, including pests and pathogens. This is a good example here. The EFB probably could be mitigated if our data shows that nutrition is compromised in those hives when they are in blueberries because blueberry doesn't produce enough pollen. Mostly the climatic conditions, like Jeff can attest to this is the conditions in April, May are not ideal for bees to forage. They're not getting enough pollen into the hive and that causes nutritional stress as well.

Jeff: Looking forward, I know there are so many beekeepers in this part of the country, and I'm sure as you pointed out in other parts of the country too, looking forward to the results of this study to help them with their annual battle with European foulbrood as they pull their bees on the blueberries. It's a very important crop for many beekeepers and for the blueberry growers.

We only have a few minutes left. If you can, and maybe we can ask this at a different time, but if there was someone, I've not asked this of anybody else who's run the bee lab, if someone was wanting to pursue an advanced degree in beekeeping, what's the process? What would be the first step and second step and third step without getting too far into details, how should they prepare their plan?

Ramesh: Looks like this is my opportunity to get some students now.

[laughter]

Ramesh: I always say you have to be passionate. You don't have to be a brilliant scientist. I don't think you become a brilliant scientist in a day. It's a process. It takes a long time to really learn. I have a couple of new students, I tell them, even I was like you. The writing, the meticulous nature of designing experiments, everything comes with experience. The first thing is you have to be very diligent and passionate about bees. Otherwise, probably, it may not be because there'll be a lot of fieldwork. People think I have some molecular biologists next to my lab here.

Their work is very different. They're always sitting mostly in the lab or bench work. Honeybees are different. You need a mix of both. You have to be passionate about being in the field. For that EFB project, we were in California for almost 10 days. We were waiting for the rain to stop some days. Everything goes differently. Biological experiments in the field are not easy. I talked about this pollen collection thing. I know we have some volunteer citizen scientists helping, but it's a painful part. You have to be ready for those. It's not an easy endeavor.

It's a commitment. You can't take graduate studies-- it's not like your undergrad where you just take classes and pass. They have to be mentally prepared for that. Then after that, yes, it's a process. You try to reach out to all the bee scientists that are across the nation and that's the easy one. You don't randomly apply to a university and hope someone will look at your application. What I advise them is look for Google or whatever your sources are. Look for the bee labs across the nation. Send them an email, not random email. Look at the profile.

I see sometimes I get some international emails. They will not even change my name. I know that this guy has sent emails to 100 more people, then I don't even read that email. It's for your audience, if they are interested, read their profile, see what type of publication they have. If they're not interesting to you, don't apply. Just look for whoever you have interest in. Maybe you are into breeding program, so don't apply to my program. I don't do any honeybee breeding. You can see and look at what your areas of interest are. Of course, nutrition is one that will be area of focus for the next 20 years as well, because there is so much to be known.

You can look for those interests and then send a short email, maybe a paragraph with your CV and your statement of interest saying that these are my interests and do you have any assistantship. I don't want them to pay and study, get a graduate degree. Most of the time we have assistantships available because we keep writing grants. Most of PIs have some sort of funding. They can ask, can I get an assistantship to study a master's or a PhD and then wait for those responses and then be persistent. Sometimes they may not read your email because they're busy.

Send a friendly reminder after two or three weeks say, can you please if you have anything? I have seen some very persistent students. They will not lose heart. They will keep trying and that tells me that they're very passionate, they really want to get in. If I have money, then I would try to take them. That's a basic process, Jeff. I know I can keep going on giving tips, but I think that that's enough.

Jeff: I appreciate that, because we talked to so many researchers. For someone who has an interest in, wants to pursue advanced studies in bees and bee biology or nutrition or wherever, we've never really discussed where do you start.

Ramesh: Age is not a factor, Jeff. I know there are people who want to do-- I did a little non-traditional not too late, but there are people who really are passionate when they are in their 40s or so as well. Again, it's all about passion and whether you have time commitment for doing that. Again, can't be just your freshly minted bachelor's degree students here but anyone, it's the same process.

Jeff: Older than 40?

Becky: I was going to say, Jeff, are you now asking about yourself?

Jeff: Older than 50 maybe?

[laughter]

Ramesh: Yes.

Becky: Ramesh, have any of your undergrads moved on from working as an undergrad student to becoming a graduate student for you?

Ramesh: Even the same lab? No. From a master's to a PhD, yes, but not from undergrad. I always encourage them to go somewhere else. I don't want them to stay at the same university unless they're really passionate about some project and that's happening only here. Most of the undergrads, I think they have gone to some other labs and pursued something non-honeybees as well. There are one or two that I can think of that have done master's and then started a PhD.

Jeff: Dr. Ramesh Sagili, we really appreciate having you on the show today. You've really enlightened us and our listeners on the importance of nutrition for honeybees. When this episode airs in December, it's getting your bees ready for the winter, and getting them ready in the springtime for the important summer, nutrition is key to that. Thank you for helping us.

Ramesh: Thank you, Jeff, Becky. Thanks for the opportunity.

Becky: Great way to spend an hour. Thank you very much.

Ramesh: I didn't realize.

Jeff: No, Becky, I am not planning going to graduate school. I was asking just in general. As I said, we've never really asked that question. Of course, I could have asked you.

Becky: It's probably better to ask somebody who actually has a lab, and who's accepting graduate students. I think you made the right choice. Honestly, the recommendation I tell people is if they are still an undergraduate, go get a job in a bee lab, because not only will you get a sense of what it's like and know whether or not this is the kind of work you want to do, but they all know each other. All those professors, they all know each other. They talk at conferences and they're going to be able to give you a nice recommendation if you want to go someplace else, or an introduction to another lab. Just get into one of those lab families. That's a good start.

Jeff: Ramesh was talking about all of the difficulties and challenges of working in the bee lab. He's talking about the hours and the fieldwork, and everything. Most people, when they think about getting into bees like that, they think, the stings. You must get a lot of stings if you're going to be a PhD in entomology and work with bees. His whole point was he didn't even mention that. He was talking about the hours and the weather. It was a good discussion.

Becky: I used to have to collect data every four hours. I'd have to go get up every four hours, drive to the bee lab, go to my observation hives, collect data, and then go back home, sleep for a little bit. When some of those experiments run, you have quite the schedule. Your field season, that is the most important part because it can delay your graduation by a whole year if you screw something up. You have to set up the genetics of your bees and the colonies. Everything, of course, isn't going to go just right, but boy, is it an adventure.

Jeff: Sounds like fun. I'm glad we had Dr. Ramesh Sagili on the show talking about nutrition and graduate work in his bee lab.

Becky: I feel like I learned so much from him, so that was great.

Jeff: That about wraps it up for this episode. Before we go, I want to encourage our listeners to follow us and rate us five stars on Apple Podcast or wherever you download and stream the show. Even better, write a review and let other beekeepers looking for a new podcast know what you like. You can get there directly from our website by clicking on the Reviews tab along the top of any web page.

We want to thank Betterbee and our regular long-time sponsors, Global Patties, Strong Microbials, and Northern Bee Books for their generous support. Finally and most importantly, we want to thank you, the Beekeeping Today Podcast listener, for joining us on this show. Feel free to leave us questions and comments on our website. We'd love to hear from you. Thanks a lot, everybody.

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