Project Apis m - Costco Scholarship Awards (S5, E11)

S5, E11 – Project Apis M - Costco Scholarship Awards

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

Kim Flottum: I'm Kim Flottum.

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Jeff: Thanks Sherry and thank you Global Patties. Each week we get to talk about how much we appreciate our sponsor's support and we know you'd rather we get right to talk about beekeeping. However, our great sponsors are critical to help making all of this happen. From the transcripts, the hosting piece, the software, the hardware, the microphones, the subscriptions, the recorders, they enable each episode.

With that, thanks to Bee Culture magazine for continuing their presenting sponsorship of this podcast. Bee Culture has been the magazine for American beekeeping since 1873. Subscribe to Bee Culture today. Hey everybody, thanks for joining us, we're really happy you're here. Before we get started, just a quick reminder to subscribe or follow Beekeeping Today Podcast and give us a five-star rating. It really does help.

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Have you listened to an episode and thought, "That person sounds really interesting. I'd like to know more about them"? Well, now you can. Each episode links to a guest profile. Each profile has a guest photo, bio, contact information, including Instagram and Twitter details if they have them. Check it out.

Finally, share the podcast with your beekeeping friends, email them links or mention it at your next beekeeper meeting. Hey, everybody, thanks for listening. We know you have choices and we are grateful Beekeeping Today Podcast is one of them. When we started this podcast back in 2018, not only did we want to bring you the known and established voices and beekeeping in honey bee research, we also set out to highlight the new voices.

Today's episode, we introduced two new researchers, Project Apis m and CostCo established a PAm-Costco Scholarship Awards in 2013. The students who receive this PhD scholarship award bring new energy, ideas, and expertise to the scientist on the leading edge of honey bee health research. The award program supports outstanding graduate students pursuing a research-based doctoral degrees in fields of enhancing honey bee health while improving crop production.

To date, Project Apis m-Costco Scholarship Program has awarded over $740,000 in the US and over a $140,000 in Canada to new researchers committed to understanding and improving honey bee health. The awards provide up to three continuous years of support for graduate education and research and $50,000 annual increments. Today's guests are two of the 2022 winners. In fact, they are co-awardees, Audrey Parish and Chris Robinson. Audrey is a PhD student in Irene Newton's lab at Indiana University. As you will soon hear, Audrey studies how the honey bee larval microbiome helps predict larvae from various stressors.

Chris is also a PhD student at Indiana University studying the ecology and evolution of a honey bee viruses using evolutionary genomic, computational biology, and classical entomology. Of particular interest and as you will soon hear, his research is exploring the relationship between the endosymbiotic bacteria Wolbachia and the Varroa Mite. Wolbachia has been demonstrated to block the transmission of viruses from mosquitoes to humans.

Chris has research is to explore whether or not it can also block the transmission of viruses from the Varroa mite to honey bees. Wow.

Put on your thinking caps and even plan to check out the episode transcript as this episode is packed full of great information from these two great award-winning researchers. First, a quick word or two from our great supporters.

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Jeff: Hey, and while you're at the Strong Microbials' site, make sure you click on and subscribe to the Hive, their regular newsletter full of interesting beekeeping facts and product updates. Hey, everybody, welcome back to the show. Sitting across the virtual Zoom table right now are the 2022 Project Apis m-CostCo Scholarship award winners, Audrey Parish and Chris Robinson. Welcome to the show, everybody.

Audrey Parish: Hi, thanks so much for having us.

Chris Robinson: Thank you for having us.

Kim: That's nice to meet you guys. I'm pretty impressed with how this has turned out and I hope you can tell us a lot about it.

Audrey: Yes, we're really excited too.

Jeff: It's a lot of work that you guys have accomplished, and working with Project Apis m, or will refer to it interchangeably with PAm, is quite an accomplishment. It's really cool, and they've always been a great resource and information for beekeepers.

Audrey: Yes, we're really honored to be working with Project Apis, especially in conjunction with CostCo because it means that we're going to have a wider audience. That's really exciting for us.

Chris: Yes, absolutely. For me, it's always nice to have something tied into an applied research output, and having both Apis and Project Apis m with Costco really, really helps me feel really good about that, the research means something.

Jeff: We'll talk more about-- Is it a contest? You call it a contest. What's the proper term?

[crosstalk]

Audrey: It feels like a contest. Yes, you can call it a grant proposal.

Jeff: When I hear contest, I think of cornholing. Anyways.

Audrey: Yes. Surprisingly, no physical challenges for this one, which is a relief.

Jeff: All right. Tell us who you are, your background, and your beginning story, if you will, how you got interested in bees and what led you to this moment.

Audrey: My name is Audrey Parish. I am originally from Houston, Texas.I went to the University of Houston, where I worked on fruit flies. Then I moved to Indiana University in 2017 to start my PhD on something completely different, working with the microbiome of honeybees. I've been doing practical beekeeping since 2018, that summer when we set up our first bee colonies. It started off with just two people and five hives. You're looking at one of the co-managers of all of our apiaries here at the lab at Indiana University. I'm not saying I'm the best beekeeper in the world, but I'm certainly learned a lot along the way.

What I do in my PhD is look at how the microbiome, that is all the bacteria and sometimes fungii and viruses, that are living inside of a honeybee colony interact with their health. I focus on the health of larvae and what the bacteria associated with larvae are doing to help them along their developmental way. If you're a larva, you're pretty sensitive.

What I've been looking into in my first chapter is how one bacterium called Bombella apis lives inside the larval diet, which you may not know, but the larval diet is a really hard place to live if you're a bacterial, super antimicrobial, but this one's special bacterium seems really uniquely able to live there. I look at how this bacterium Bombella apis interacts with the diet and changes it, increasing its nutritional content and what the long-term ramifications are for the larvae.

Chris: Hello, everyone. My name is Chris Robinson. I just finished my first year of my PhD at Indiana University with Irene Newton and Curt Lively. These are my advisors. I'm from South Carolina, did my undergrad at the University of Massachusetts, Amherst, graduated in 2015. Then I wandered around for about six years figuring out how to get back to grad school and what exactly I was interested in.

I worked for the USDA, I worked in restaurant kitchens in Charleston, I was in Chornobyl for a Fulbright fellowship. Looking into it, I found everything I was doing was linked to food. Especially in Ukraine, I was working there for a year. I went to a bunch of native beekeepers in the Carpathia mountains and saw their operations and saw how they incorporated all these different elements of beekeeping that they've been doing for, I don't know, five, six, seven generations. It really opened up to me a whole wealth of questions that I wanted to ask.

I pursued that system, applied to grad school like three times, kept getting rejected, and finally, reoriented my questions in the system, "How do I ask cool questions in bees?" and really lucked out and ended up here at Indiana University. Fundamentally, right now I'm really interested in these things you may consider, I guess, mobile genetic elements or little pieces of DNA that move around, these things that act like life. They replicate, they make more of themselves, but they're not technically living in the sense that you'd say a cell or a bee is living, but they do things that are unique to living systems.

Things like viruses, things like plasmids, if you've tour of those, transposons jumping genes, I'm interested in all these different things. This is important because in particular in bees, bees are dealing with this massive pandemic, I should say, of viral diseases, especially exacerbated by Varroa mites. I'm really, really interested in understanding the fundamental evolutionary ecological processes that make certain diseases worse than others or what causes a certain pandemic or not.

Jeff: At some point during the conversation, I do want to hear about your time in Chornobyl.

Chris: Oh, sure. Anytime it's fun to talk about.

Jeff: Very timely right now, too.

Kim: Chris, let me back up a half a sip. When you were overseas, that's where you actually began working with bees, you weren't doing something here first?

Chris: When I was at the USDA Agriculture Research Service on a watermelon research farm in South Carolina, there's a group of master gardeners that would come in and work with Clemson extension, who we shared facilities with, and they would train master gardeners. I would work with them. I was a plant pathologist at the time. We set up a bunch of bee colonies that we would just have to help pollinating the flowers and help manage resources on the farm.

I worked a little bit there with cleaning frames a little bit and just general management, moving things back and forth, but I really didn't get a deep agricultural lesson. The person who was working that was very protective, but in Ukraine, it was very different. The way that these villages interacted with these hives was incredible. It was the first time I could go up to a hive and just see someone pull out a frame and just work with these bees and everything was totally docile, incredibly calm. They could do anything they wanted to do with the bees.

That's when I really, really fell in love with the system and saw it as this really, really beautiful symbiosis between human and insect. That's really deep in my enthusiasm for it, but before that, I really had no exposure. I grew up around farms and no bees at all.

Kim: Are you now working with Audrey in the bee yards?

Chris: I am, yes. She's shown me a few times now. My first couple of weeks here, I went to the University of Illinois and Audrey helped me collect bee bread from their very, very nice bee yard. That was beautiful. Then this past fall, I went and collected mites from a few different beekeepers to start doing a general survey of deformed wing virus infection across Bloomington, Indiana. Then this past summer Audrey showed me around. Before the summer's over, I'm going to do some larval rearing and feed them and raise them from incubation.

Jeff: I'm sorry, what was that word, larval rearing?

Chris: Larval rearing. Audrey's probably the world expert on this. Really, really beautiful to see.

Jeff: Collecting bee bread out of a cell takes a technique.

Kim: Fussy work. It's hard to do.

Jeff: Yes, it is.

Chris: Oh, yes. I tore up that whole frame. I tore it big time.

Audrey: Bees will fix it.

Chris: I know.

Jeff: Especially when you get mad at it and throw it across the bee yard.

Chris: That's what I liked about it. It's like they can feel the energy of the room. If you're in there getting angry, they're going to get angry. You hear that buzz go up in frequency. I learned with Audrey really quick that you got to enter this with cool, calm, collected energy. Don't let any of your bad vibes flow out, which was nice.

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Jeff: Like you, one of the first things I learned is that to be a good presence in the bee yard, you have to totally clear your mind of everything else and just be with the bees. Then it's a lot better because, like you said, they sense everything. That's fantastic. I'm glad you noticed it.

Kim: It'll save you a lot of pain down the road. Sometimes it's hard to do when you've got 10 minutes to do a 20-minute job and you gotta get it done. Then it all goes out the window and you hope you have enough armor on.

Audrey: You listen for that downshift in vibration and you know you're in trouble.

Kim: Yes. What will your work do to help bees and help beekeepers if you get a bigger picture there? Because I know it's a pretty specific topic both of you have, but will this eventually, along with other discoveries that people are chasing right now, make bees and beekeeping better?

Audrey: Yes, I absolutely think it can, especially when you're looking at the larval microbiota. I think there are a lot of really wonderful probiotic products out right now that focus on the adult bee, and while those are really important for sublimating a colony, those may not be the bacterial members that are going to do the best for the really vulnerable larvae. We know that it's really easy to supplement Bombella apis into sugar water and we know that it can survive in the niches where it's supposed to be. One thing that we're looking into, not just in the lab, but how we can apply this to beekeepers because we've surveyed a lot of beekeepers, so we know that their most valuable resource is their time.

We would never propose something to help bees, even though we know we have the science behind it, that means you have to go in every single one of your 200 colonies and spend 30 minutes per week on them. One thing we're really excited about are the probiotic supplements like Bombella apis, and like these Wolbachia and mite treatments is that they're naturally associated with bees. One could essentially plug and play. You could put this bacterial supplement in some sugar water and let the bees take over. We're really excited about the ease of our long-reaching applications.

Kim: When I'm feeding, all I'm going to be doing is feeding. I will have added some of your discovery to that feeding pale. Then I just get out of the way. I'll let the bees feed it to the larva and the larva then are going to benefit from having either some of this or maybe more of this in their diet?

Audrey: Precisely, or just the right strains. Say, we were starving and we had to give you the best supplement that we could, we would make sure that we're screening because there's a lot of diversity, even in just one species of bacteria. From one larva to another, they could have really different strains. It's important to have a superpowered strain that's producing a lot of the things that you want it to be producing. It would be as easy as delivering that in the sugar water. We know that the ones we're working with can hang out for about 72 hours. Yes, it would essentially be like mixing instant coffee into a cup of water, except you're mixing a probiotic into your sugar water for your bees.

Jeff: I will jump in here right now and say that one of our sponsors, Strong Microbials have just released what they call Extend, and it's a water soluble probiotic.

Audrey: Yes, we are not reinventing the wheel at this. There are good people already on some of the same tracks, just with different bacteria.

Kim: This bacteria is going to be raised in bulk someplace by somebody like Jeff just mentioned, and I as a beekeeper, I'm going to buy a package of it at the bee supply store, dump it in my sugar water. Right now, do you have a feel for the cost of this?

Audrey: Yes, that's a great question. It can't be extremely expensive because I have engineered a diet that these bacteria grows in, which is essentially just some cheap vitamins and a couple of different types of sugar and some yeast extract. The process of growing it is really simple. It doesn't require special chambers, it likes oxygen, you just have to shake it up and let it go for 24 hours. The cost would be pretty minimal from the production side.

You would just need to dry it out and get it into a deliverable form. I can't imagine that it would be terribly expensive. We know that, again, just like your time, no one's going to spend $20 per week just to improve the health of your larvae. I understand that that is a consideration.

Kim: To me, looking at this from the outside, it begs the question, once I have put this in my beehive, do I need to do this every week, every month, every year?

Audrey: That's an excellent question. I wish we had the production up and running so I could answer questions like that, but that would be something that we'd really like to look into in this upcoming season.

Jeff: Did you look into the seasonal differences in the larval requirements?

Audrey: Yes. There aren't going to be larvae in the winter, while they over winter and the queen's not laying, and so there's not going be any need to supplement in the winter. If I had to guess, I would say the very best time to supplement is going to be early in the season when you're getting this larvae up and running because you're producing the workers that are going to jumpstart your colony in early season.

If I were to dream up a perfect scheme of supplementing with Bombella apis, it would be probably March through May and then let these that have already established, which is one great thing about using a natural bacteria that's already associated with colonies, is it's going to establish pretty easily, let the established take and run with it. Then don't worry about it in winter because you don't have the larvae.

Kim: There are places that will probably need it year round.

Audrey: Exactly. That's quite true. What's so interesting is, in the parts of the country that have different seasonalities, it could be that you have plenty of pollen because what we want to supplement is the protein that they're getting and the bacteria help with the protein content of their diet. It could be that maybe if you are a beekeeper in Illinois, you want to supplement in the spring to get your colony up off the ground, but it could be if you're a beekeeper, say, in Texas, you'd rather supplement in the fall and there's going to be less pollen available. It could be something that is different depending on where you are.

Kim: I can see a booklet of instructions coming with this.

laughter

Audrey: I'll just refer them to the podcast.

Kim: Chris, what's your role in this?

Chris: My role is very different. I came in and saw this great work, saw Audrey and another graduate student, Delaney, who's also working on Bombella, have built this system and talking with them. I was like, "They're approaching this from understanding these probiotic, these beneficial ones. I want to try to develop some understanding about what's the diseases that are affecting bees, and maybe the combination of those two things can really, really help dampen some of the effect."

Out of my project, I'm hoping to make two goals. The first one is a little less tangible, but I think incredibly important. This is just to understand some of the fundamental processes that drive the evolution of viral pathogens. What makes them more virulent or be mortality? What causes them to transmit easier? What does having Varroa there, Varroa mite feeding on a bee and transmit it to another bee, how does that change what viral strains are in a given viral population? On top of this, how does the presence or absence of Bombella, the bacteria Audrey is working with, effect that?

All these different things, all these different interactions, I'm really curious in trying to understand the basis of-- The idea being is something I learned a lot as a plant pathologist is that we spend so much time reacting to pathogens and pathogen pressure. That takes up so much time and so much resources that it becomes really, really difficult to try to think of how to get ahead of it.

I think working to make a deeper understanding of the evolution in ecology of bee viruses helps you really try to think about how to get ahead of it, what's important in the environment. Does having Varroa there or not really make a difference? Talking with beekeepers, the virus is not a problem, it's the Varroa mite in conjunction with the viruses that are a problem.

That led to the second fork of my project, which requires some context. Irene Newton is a world expert in cell biology of this cellular symbiont called Wolbachia. Wolbachia is probably one of the most successful symbiotic organisms in the world and infects 40% of all known insects. It's maternal transmitted. Every daughter or every mother gives it to her daughter under right conditions. It spreads incredibly fast. Beyond all the interesting biology, it has this really, really interesting effect, where if a mosquito, for instance, is infected with Wolbachia, it then cannot generate viruses that cause diseases of humans.

If I have a mosquito that has Wolbachia, cannot get Zika, cannot get dengue, all these human transmissible viruses, it can't spread it. It has this pathogen-blocking effect. Reading the literature, I saw that Varroa mite, which we know is a major vector of bee viruses, there is some, I guess, variability on the different Varroa mite that if they do or do not have Wolbachia, but it's currently completely unknown if Wolbachia is persisting in Varroa mite or not.

Another part of my project is to go out and do this survey of Varroa mite to see if they actually have Wolbachia, document their persistence, their demographics of Wolbachia in Varroa mite, and then actually do a fine tune analysis just to see if having Wolbachia in Varroa mite has a pathogen-blocking effect. Does having Wolbachia there actually block viruses from happening or the transmission of viruses?

The idea being that maybe out of all of this, the dream finding would be that if you've got Varroa mite infected with Wolbachia, it may feed on your bees, it may cause some stress, but the presence of this symbiont means that they won't be transmitting viruses. Having talking with beekeepers, that would be the best thing that could happen. At current apiculture practices, you're not going to get rid of Varroa mite. Unless you completely change the way that commercial beekeepings going, you're not going to get rid of it. It's just going to keep evolving, being resistant, these highly dense hives. If you can't get rid of it, if you can block the transmission of its viral diseases, I think that would be a massive way for beekeeping.

Kim: What I see happening here is you are applying this Wolbachia to a hive to be consumed by bees that are then fed upon by mites. That Wolbachia then goes to the mite and the mite is blocked. Then?

Chris: A mite that would be infected with Wolbachia would be introduced to maybe an already existing Varroa mite population. Then that Varroa mite with Wolbachia would mate with other Varroa. In the particulars of Wolbachia biology, it can force itself-- If it mates with something that it isn't infected with Wolbachia, it's offspring die. It ensures that only Wolbachia-positive offspring come out. Really, really cool. In a very, very short amount of time, you would get populations of Varroa mite that are only Wolbachia-infected. Hopefully those Wolbachia-infected Varroa mites wouldn't be able to transmit viruses because of this Wolbachia infection.

Jeff: I can just see it now, in the bee catalog a coupon to order a thousand Wolbachia-infected Varroa mites. What a paradigm shift, selling-

Chris: That was one of the things that a Audrey and I were worried about because it seems like the solution coming at us, it's like, "You want us to go and put Wolbachia or Varroa mite in our bee colonies? That's insane." If the Varroa mite is already there and you're just introducing one that can't transmit viruses-

Jeff: They'll be asking what you're smoking there in school.

[laughter]

Chris: I know. It's a crazy idea.

Jeff: No, I think it's actually a very cool idea. I'm having fun with it, but it is very exciting work.

Chris: Oh, no. I think it's got a lot of potential and it's really unexplored, which is really nice. As a grad student coming in, trying to figure out what you want to do, some things, emergence a really, really simple like, "Let's just see what happens," very exploratory. I'm sure beekeepers will love it if we can come up with some cute Wolbachia brand name for the Varroa mite.

Audrey: [laughs] Or gentle mites.

Jeff: Wolbacha, yes.

Kim: I'll go back to the question, is how do those people that I'm going to buy these mites from get those mites infected in the first place?

Chris: That would require the developed research to develop a system to maintain Varroa mite in vivo, either in the wild or something, maybe having a big bee yard, where you're constantly having ongoing generations of Varroa mite feeding on different colonies. Then, just as long as you've got Wolabachia already in the population, it's going to persist. I can imagine something ongoing, just random sampling of your Varroa mite in your colony. You could be a quality control to ensure that you still got ongoing Wolbachia infection.

Jeff: How does the Wolbachia vector between one Varroa to the other? Is it contact? You said it was maternally transferred, but how is it-

Kim: How did mom get it in the first place?

Jeff: How did Eve Varroa get it?

Chris: I would imagine that they could probably get it from bees or anything that they feed on. I'm not entirely sure how maybe the initial- [crosstalk]

Audrey: We're touching on a gap in the literature.

Chris: I really have no idea. I don't know that there's-- There's actually not a lot of work looking at Wolbachia and honeybees either, so maybe they could get it from feeding on bees, but there's already Varroa mite existing that have Wolbachia, so it would just be a matter of finding one and using that to seed a new generation, after you do a lot of the research to figure out which Wolbachia strain's the best, which one has the best pathogen.

What's really, really nice about this approach is that because it's biological, these viruses, as far as we know, they can't evolve, escape this effect. As long as Wolbachia is there, it's not going to evolve resistance to Wolbachia. Wolbachia is just really good at persisting and maintaining this phenotype.

Jeff: I can see why that is exciting for you. The whole area, as you mentioned, but briefly, how it opens up a whole field in area of research, the Wolbachia gap to how do you commercialize it? How you distribute it in an apiary? There's a lifetime there.

Chris: Oh, absolutely.

Jeff: Very cool.

Kim: No poison in my beehive. That's the top of the pile there. I'm not putting poison in my beehive [crosstalk] anymore.

Audrey: Exactly.

Chris: That's one of the things working with Audrey and noticing talking about how little work goes into maintaining these beehives, ideally. Just trying to minimize that as a scientist. People are busy, I want to make sure that whatever solution they come up with is one that's cheap and one that they don't have to do any time. Something that's self-persisting, this kind of feedback is ideal.

Jeff: This is a interconnection between your two research, is that Wolbachia in the larval gut biome.

Audrey: Even a larger umbrella than that, is the microbiome, whether it be Wolbachia, just the general colony. Wolbachia in a Varroa mite or the bacteria in the bees of the larval diet, this bigger umbrella being blocking pathogens via naturally associated symbiocies you leverage what's already there without adding something that could be potentially harmful in ways that we don't understand yet. The big umbrella is protecting from stressors with these natural associations.

Jeff: [chuckles] Very cool.

Kim: Well, I can see then that working with these two organisms are going to be beneficial to the colony. The question that just came up was, can I feed my bees both of these bacterias, that it's going to aid the larva, and if the larva is eating both of these bacteria, is able to eat both of these bacteria, it's going to protect them, and a mite feeds on that larva, a mite goes into cell when it's kept and feeds on that larva, then both will have your bacteria. When it emerges, she'll be unable to transmit any more viruses. The larva will be healthy because of one bacteria and be able to spread the other bacteria so that the mites that emerge can't continue to spread viruses in the hive.

Audrey: That's exactly it. Yes, we build up the larvae with these natural probiotics, we turn perhaps a protein deficient diet into something that is richer and healthier for them, and then we essentially pass the baton as soon as the pupa is capped, and now these Varroa mites which are deficient in transmitting viruses, while they might feed on a larva, it should be healthy enough to withstand that. The next maggot or pupa that that Varroa feeds on isn't going to be transmitting viruses.

Kim: I'll take a quote of this right now.

Audrey: [laughs] We'd be very happy to be in production.

Chris: I do want to clarify that the Varroa mite, it wouldn't transmit the Bombella.

Audrey: It's true. The Bombella is hanging out in the larval period.

Chris: [crosstalk] but I don't think it does.

Audrey: The bee doesn't have the Wolbachia, so they're separated into different organisms.

Jeff: Right, but Bombella would then spread by all the other bees and the shared nutrients and then Varroa mite, it would have eaten, they would have parasitized off a bee, would have offspring, and all those offspring would have-- It already makes sense, if I'm a varroa mite and I'm causing disease in my colony, it might be better in the long run if I'm a Varroa mite, I can parasitize off bees, but I'm not actually killing them with any viral diseases.

That would be true in every case. There's some math you can do to actually figure that out, but for the most part, that seems to hold true. If you can remove that viral pressure, bees don't really seem to mind.

Kim: You come to a healthy plateau of all of the populations of viruses, mites, and bees in your hive. Everybody's happy and nobody is killing anybody else.

Audrey: Everyone's medium happy. [laughs]

Chris: What's really cool is these viruses are there already. This deformed wing virus... they're already there, this kind of base level equilibriums that the bee is happy with, and having these mites there is what really messes with the equilibrium and causes these populations to act all different. Yes, finding a good happy place immediately with everybody without getting rid of anyone, you're not going to eradicate living things, they will always find a way to survive, I think it's the best solution. As a researcher, it's what I'm trying to get-- I'd love to see agriculture move more towards that harmony with living things.

Jeff: If this all goes well and based on the awards and scholarships and the co-presenter of the Costco, I'm sure all this will be available at Costco quantities too.

Audrey: Yes. Isn't that wonderful? We've got a direct plug into a major retailer. I'm sure they'd love that, so would we.

Jeff: There you go. Well, this is really, really exciting. What's the next steps?

Audrey: I have one small vignette that I'll leave with on this project before I talk about my next steps, is our umbrella is the viral vectors, and I will say that Bombella apis doesn't just supplement nutrition, it looks like. I won't give too much away because there will be a paper on this eventually. I'd like to keep it a little closed, but it turns out, I think as beekeepers, we're always hearing about synergistic stressors.

What's the problem with colony collapse? Multiple stressors, and we know that nutrition is one of the leading stressors and so is viral pathogenesis. We know that when honeybees are nutritionally stressed, it's easier for viruses to come in and hurt them. The next step of the project with Bombella apis is looking into how this nutritional supplementation, this buffering of larval health helps protect them against the viral pathogens, specifically Sacbrood virus.

I'm sure you guys are familiar with Sacbrood. It essentially turns the larva into a brown water balloon, it's not exactly ideal, and it's, interestingly, propagating the same niches where you find Bombella apis, so it's in the diet, it's in the nose glands that produce royal jelly, it's in the larvae themselves, it's right there alongside Bombella. We're really curious how these two might interact and whether the presence of Bombella can protect larvae.

Without giving it fully away, it looks like in some preliminary studies, just the presence of Bombella, either through supplementation of nutrients and strengthening the larva or a direct interaction, which would not be unheard of, it actually improves the survival when challenged by Sacbrood by about 25%. The next steps are looking beyond the nutritional supplementation of Bombella apis and seeing how it self-protects against some of these interesting larval pathogens like Sacbrood virus.

Jeff: Wow. That's exciting.

Audrey: It's a silver bullet. Without promising too much, we'll have to do some larger experiments and include some other viral pathogens and track them along their lifespan, but it looks like we have an interesting silver bullet. It's kind of skated under the radar in the microbiome world.

Kim: All of this work, and it may be some extraordinary results here, is coming from the result of a grant provided by Costco, and if I have this right, your project received $150,000 from Costco to fund this work?

Audrey: Yes, that's true. That's absolutely right.

Kim: It says a lot for Costco, in my opinion.

Audrey: Yes, we're extremely grateful and we've been working with Project Apis m over the years and Costco is their number one supporter, and so it's really wonderful to get to work with both of these entities.

Jeff: Absolutely. Thank you Costco.

Chris: That said, not to harp on it too much, but I really admire Costco's approach to research. I know when they wanted the rotisserie chickens they sell there, they couldn't find the actual right size of chicken to have for their rotisserie chicken, to have that kind of right weight size, so they just funded a bunch of basic research and animal breeding to actually find the research for Costco-sized rotisserie chicken, which I think is really, really cool. They enjoy the science, it seems like it.

Kim: I guess they want to keep honey on their shelf too, right?

Audrey: Exactly, and not to mention fruits and vegetables and all those other trifles that we might want to keep around in the future.

Chris: Yes, those niceties.

[laughter]

Jeff: I'll let you start off in alphabetical order. What's next?

Chris: For me, it's a bit different. I'm probably at the opposite stage of where Audrey is at. I've got a lot of the peculiarities of a PhD program to deal with, prelims, these big exams, and stuff, but what's coming next for me is to go out and basically survey beekeepers across Southern Indiana and see, okay, which one of these hives are infested with Varroa mite? Collect Varroa mite and then do some simple presence or absence screening to verifying do you or do you not have Wolbachia infection with Varroa mite? Just get a good idea of the ground-level Wolbachia infestations or infections.

With that, you can really start to kind of build, "Okay, is this prevalent? Do we already see some demographics?" We'll take some recordings. If I find a colony that has a lot of Wolbachia and a colony that doesn't have a lot of Wolbachia but they both have Varroa mite and it turns out that one's maybe healthier than the other, maybe there may be some pattern that emerges just from looking for this step during this kind of survey.

That's step number one, go ahead and drown the actual field that I'll be surveying, and then as that's going along, also work-- What I've been working on through my first year now is learning the computational tools necessary just to really interrogate and analyze these massive populations of viruses. How do you tell one virus apart from the other? How do you determine one that's going to cause disease versus not cause disease? Severity? Is one strain of virus worse than the other? Is that strain present? Figure out how to characterize all of that. The next step would be to combine the two and say, okay, where I see Wolbachia infecting Varroa mite, do these take bees from where there's Varroa mite but no Wolbachia and then use those computational tools to then ask, these viral populations any different from one another? Is there anything going on in their interaction that may be a result that could say, "Wolbachia is causing this"? Then hopefully, all that combined will give us some indication if Wolbachia's presence has some sort of effect on viral population dynamics.

Jeff: Let me ask real quick, Wolbachia, can you sample a hive and determine whether there's Wolbachia in a hive or do you have to collect so many Varroa and test each one and put them in a blender?

Chris: Yes, you'd go out, and Audrey step in at any time because I've actually never done this, but you'd go out. We'd have to think hard about a correct sampling strategy, whether we want to do it by Varroa mite. You could certainly do it individual Varroa mites that you collect from colony. You can pull them if you're trying to save time, but maybe these most sensitive ones, you may go out and collect, say, 20 or so Varroa mites from each colony, you'd grind each one individually, which doesn't take super long, and then you would extract-- Just put them together and it comes out.

You'd extract some DNA and then you may want something like a PCR gel, which looks for a specific sequence that you're interested in, or a western blot, which would look for a certain protein. Wolbachia has characteristic genes and proteins that we could use to tease out. If it's there, you would see a really, really bright big band on what looks like a gel or like a piece of Jell-O. A band being there will tell you, yes, Wolbachia is there, and so we could use that as a cheap, easy way to go ahead and interrogate those demographics.

Kim: You have to start with live mites?

Chris: Or freshly dead.

Kim: Freshly dead.

Chris: Yes. Ideally, live mites. If I could get there right as they fall on the mite paper or the mite pad and collect them there, that could also work, but we have no idea how long Wolbachia would persist.

Kim: That'd be a good question. Can I do an alcohol wash and pick up a bunch of mites all at once very quickly, do the wash, separate the wash from the mites, put the mites in a jar, take them back to the lab and do the things that you want? They're going to be dead, but freshly dead, as you put it.

Chris: That would totally work, I think.

Kim: The next question could be then, if I were you, how long can those mites be dead and me still get an accurate reading? Because I can see you taking samples from all over the US on this and you're looking at different strains of virus. Are they different in the Southwest than the Northeast? That sort of thing. If I take a sample today, I put it in UPS and overnight it to you, is that going to work? I can see that being the next step.

Chris: Absolutely. There's so much tantalizing little basic stuff, so the questions that need to be answered. I don't mean basic for anyone listening. We use that word a lot. It doesn't mean it's simple or anything, it just means foundational stuff. There's so much just things that we know that we need to know or these unknown knowns that have to be answered. Yes, figuring out how long can a mite be dead before we can get Wolbachia out of it is an essential question.

Maybe something we can try next season, find a mite and then just trace maybe the life history of one. A big part of this is going to have to figure out how I can maybe build or rear Varroa mite in the lab, which sounds terrifying, but it might be a good way to answer that question.

Jeff: I can tell you how to do that in a lab because I got a bunch of it in the backfield. It grows just-- No, I'm just kidding. There is so many questions that come to mind, but we don't have enough time. Is there anything that we haven't asked that you would like to say before we wrap it up here?

Audrey: Ooh, that's a good question. The one question that I get asked a lot is, "Well, what can I do? How can I protect my bees or how can I protect the microbes that are associated with bees?" I say one thing that can be really challenging in working with these microbes is how much antibiotics the colony gets treated with. It can cause dysbiosis, it can throw things off and create resistance and things that you don't want. Anytime you can avoid doing things like antibiotic treating your honeybees is ideal.

Anytime you can do things, for just our most basic listeners, like avoiding roundup around your colonies would be ideal. Gentle beekeeping, like we said, they can tell your vibe, be respectful of them and try not to treat them with too many exogenous chemicals. We've got a microbe-minded being on the way for you. Please just be gentle.

Chris: I can't think of any questions, unfortunately. I just do want to say it's a pleasure to speak on this. What I really, really have loved about finishing my first year of my PhD is just how wonderful the beekeeping community has been, and how well the researchers who do beekeeping science, how they're, hopefully, constant interacting with the beekeepers themselves or the people working on their bee yards, and having this kind of back and forth conversation where you're always knowing what beekeepers need and what kind of solutions you need to drive.

I can say, as a scientist, it makes my job so much easier and so much more rewarding, so I guess a big thank you to all the beekeepers out there who work with us on answering these questions.

Audrey: I totally agree. It's such a pleasure working in a system where you have this continuum from big industry-scale beekeepers to people who just started their hobbies this year, and we all connect over something that we really love and care about. We might have totally different approaches, but we all care about honeybee health, and there's always such an open dialogue. It's such a pleasure to interact with this community.

Kim: Well, I can see that when you guys have collected enough questions that you need answering, you have to come back and share those questions with all of our listeners so maybe we can continue to help.

Audrey: That would be wonderful. We look forward to it.

Jeff: We look forward to having you back. Thanks a lot for joining us today.

Audrey: Thank you so much. So happy to be here.

Chris: Thank you both.

Jeff: I really enjoyed having Audrey and Chris on the show. It's exciting to hear about the work that they're doing. It's fun to feel the excitement they have. We've never had anybody on the show talking about either of those. Well, the biome we have, but not the virus research, Wolbachia.

Kim: I hope we live long enough to see this work. It's going to be great not to put more poison in my beehive and to have better-fed bees and not an artificial diet. Their environment will allow them to eat more better and to deal with some sort of standardized population of mites so everybody's living together friendly and happy and no poison.

Jeff: Like Chris said, they find that equilibrium where everything's coexisting. That's one of the big things about the pest, Varroa, is that it's an unnatural pest and it destroys its own host. That's not a good relationship there.

Kim: No, these guys were good. I'm glad we were able to talk to them today.

Jeff: Yes, thanks to Costco and to Project Apis m for doing these scholarship programs that you're providing. Well, that about wraps it up for this episode. Before we go, I want to encourage our listeners to rate us five stars on Apple Podcast, wherever you download and stream the show, your vote helps other beekeepers find us quicker. 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 reviews along the top of any web page.

As always, we thank Bee Culture, the magazine for American beekeeping for their continued support of Beekeeping Today Podcast. We want to thank our regular episode sponsor, Global Patties. Check them out at globalpatties.com. Thanks to Strong Microbials for their support of this podcast. Check out their probiotic line at strongmicrobials.com. We want to thank Betterbee for their longtime support, check out all their great beekeeping supplies at betterbee.com. Thanks to Northern Bee Books for their support of bee books, old new with Kim Flottum. Check out all of their books at northernbeebooks.co.uk.

Finally, and most importantly, we want to thank you, the Beekeeping Today Podcast listener for joining us on the show. Feel free to leave us comments and questions at "leave a comment" section under each episode on the website. We'd love to hear from you. Thanks a lot, everybody.

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