Human Practices

BeeVAX: […] We would like to start right away and first ask if you could introduce yourself and explain to us a little bit how far you were involved or included in the processes of veterinary medicine for honey bees.

Dr. Maus: Yes, I'd love to. As you know, my name is Christian Maus. I am an entomologist by profession. I have been working at Bayer for 23 years now. I am mainly concerned with environmental issues, including pollinators and pollinator safety in agriculture. In the years up to 2019, I was the scientific director of our Bee Care Center for a long time. That was an institution that made a connection between our bee or pollination activities in the field of veterinary medicine and agriculture. So, those from the former area of animal health controlled, and there we also had some varroacid, for example, in the population, also new products developed, and as I said, to bring the whole thing a bit holistically under one roof, the Bee Care Center was established. That was resumed after we divested the animal health area in 2019 or the following years. And that's why we're no longer working in this field. In the years where we worked closely with animal health, that was my contact point with the field of honey bee health.

BeeVAX: […] We would like to know how involved you were in the research, or rather in the sales?

Dr. Maus: We did some cooperation studies with external research partners on the topic of bee health, on the topic of Varroa control, for example, so for example research on other topics that belonged to the beekeeping practice, or how I think my bees are healthy, the question, for example, but we were also involved in the development of a new product, that is no longer a new product, which was called the Varroa Gate. That was a plastic strip with punched holes that were attached to the flight hole and attached with different varroatids that you had to rotate annually to prevent resistance, where the bees, when they were going in and out of the hive, got a small dose of varroatid. That was something where we were actually able to develop one of these gates with flumetrin. The other two could not be developed there because, as I said, we stopped the business. But that was also a concrete development work where I was involved.

BeeVAX: [...] You also talked about Varroa-resistant strains. Are you talking about genetically modified organisms, or do you talk about natural recombination that should have led to this resistance?

Dr. Maus: Exactly, it was not about genetically modified genes. We had an investigation with the University of Wageningen, where they conducted a kind of resistance breeding program, which we also supported. We also supported a scientist who apparently examined Varroa-tolerant tribes and wanted to find out what this Varroa-tolerance is based on.

BeeVAX: [explanation of our project]

Dr. Maus: That definitely sounds like an exciting approach. And I wouldn't know if they actually tried to develop it commercially. I don't know if there was research for it. We didn't do any research with entomopathogenic fungi. I don't know how it looks with others, but I don't think that anything has ever been developed close to the market.

BeeVAX: Do you think it is an approach that could have a future as a product, further developed beyond this project?

Dr. Maus: It depends on two aspects, which you should look at later, which I can't answer right now. But these are questions that you encounter with any kind of Varroa product. That's why it's good to look at them. The first is, could there be possible side effects? Could any other organism be affected by the treatment? That would have to be analyzed, in whatever form. And then again, the at least equally important and often more difficult point. The big problem with the development of a varroatid is ultimately the return on investment. The development and approval of a veterinary medicine product is relatively expensive compared to other veterinary medicine products. What you get when you put it on the market is relatively limited. So hardly a beekeeper will pay a very high price for a Varroa drug. That's why there are so many medications, for horses, dogs, or cats, where the owners of the animals are willing to pay higher amounts. With Varroa, the big hurdle is to make it cost-effective. I don't know if you can really give a value to that. I would estimate that, from beekeeper to beekeeper, more than 10-20 € per year will probably be too expensive. Because many farmers work, in addition to the conventional synthetic products, also with organic acids, which are cheap. Sure, you have to be able to deal with that, and not everyone can. But to really bring enough to the market to make it a success, it would have to be relatively cheap to offer.

BeeVAX: Yes, we thought that two advantages of our product are that it could be much better for the environment if you put it on the market. Besides that, it could be much easier in the application. [...]. Our basic idea was to vaccinate the queen with Vitellogenin and ensure herd immunity. This would be easier to use, but it would require some practice because it is not so easy to vaccinate a bee. We came up with the idea that the best way to use it would be to genetically modify the Vitellogenin protein. Because of German laws, there is a lot of thinking about the future. And a third possibility, which we had considered, would be to spray the Vitellogenin on the beehive. Here the problem would be that studies have shown that the social behavior of workers is influenced by the Vitellogenin and with higher concentrations, higher aggressiveness is correlated. And that would be less advantageous.

Dr. Maus: Well, that will certainly lead to a kind of disruption, and then the death rate will increase, but it can also be that a hive recovers quickly from it. The question is how to treat a whole hive with a spray treatment so that the large majority of the bees not only get something, but also everyone gets about the same amount. It is very difficult to distribute. Probably it would be something to talk about to vaccinate the queen if that works like that. The question is who would do that? Would the single beekeeper do that? Or would the queen breeder do that? Where would that be introduced?

BeeVAX: Probably the beekeeper should do that because the effect doesn't last forever if you just put the protein dose in place.

Dr. Maus: That's an important question. How long does the effect last?

BeeVAX: A certain period of time, but unfortunately not forever. So, that's just the way it is, the offspring that lays, then for a certain period of time, I think that was for a few weeks, but of course you would have to repeat it for the next year or maybe even twice a year.

Dr. Maus: Then the question is when to do it. You would probably have to do it at the sensitive times of the year when you want to reduce the Varroa pressure.

BeeVAX: We also thought about feeding it, but we were afraid that it could be digested. Maybe you have ideas?

Dr. Maus: Not really because I have no idea how stable it actually is and how far it will go through the digestive tract or not.

BeeVAX: We read studies that state that [Vitellogenin] will be digested. Our idea or our approach would be, [...], to encapsulate the Vitellogenin, even if it is relatively large so that it cannot be digested by the enzymes and as a result maybe be absorbed into the hemolymph. Do you know any possibilities or have you ever tried this kind of thing?

Dr. Maus: No, I don't think we have done anything with it. Yes, there are these micro-encapsulation options and that can certainly work, but we don't have any experience with that. I don't know if something like that has ever been done with bees.

BeeVAX: And now to come back to the application, [...] what do you think would be possible for the user? What could you imagine for an application that would be possible with our product?

Dr. Maus: I would recommend thinking about the geographical framework. In Germany, we have mostly hobby beekeepers. I think the average beekeeper has around 8 hives. Not in all parts of the beekeeping industry are there any innovations. There is one fraction that is very progressive, others are more on the traditional side and prefer to treat with organic acids. That can also work very well. Some innovations, I think, would be more likely to take place in markets where we have a more pronounced hobby beekeeping scene. For example, in the USA, where small-scale beekeeping starts with a few hundred hives and not with two or three. Large-scale beekeepers have thousands, tens of thousands of hives. In the USA, beekeeping is mainly for crop pollination. And there, more bees are used as a pollination aid than for honey production. And there you are also much more interested in developing innovative methods. We also have a completely different mentality than here. So, I would at least take a look at the US market. Approval is also perhaps a little less complex than in Europe. So that would be a thought not to specifically focus on Germany because I'm not sure if the beekeeping structure would support such a new product as much as a scenario where we have many professional beekeepers.

BeeVAX: In which areas would Bayer have done something with entomopathogenic fungi? Do they know anything?

Dr. Maus: We have a natural products line where we produce crop protection products derived directly from natural products, so that would have been in the range. However, I was not involved in detail and therefore cannot say much about the mechanisms of action. Normally, however, mechanisms of action tend to be more complex. But that varies from case to case.

BeeVAX: There also seems to be a competition between wild and honey bees. Could you imagine what could happen?

Dr. Maus: I don't know how far this is being investigated, how exactly one would incorporate them into the bee population, and whether other organisms could be affected by this. That would be something that has to be investigated, of course, also for the approval of the remedy. There must logically also be possible environmental effects investigated.

BeeVAX: Yes, if you talk about how that could affect other organisms, do you mean other organisms that also live in a wild bee population or organisms outside of the hive?

Dr. Maus: It is important how far that could come out of the bee hive and whether it could have any effects. In principle, other organisms that still live in the hive are either other pests, the little cloud beetle, wax moth, or just some commensals, but I think the concerns are more about what happens when the drug gets into the environment.

BeeVAX: Although, as far as I know, it is true that these traces are omnipresent, right? The traces of entomopathogenic fungi are still almost everywhere. Could the concentration of these spores be a relevant factor?

Dr. Maus: Exactly. That's usually the key point. And normally, even if we have a plant-based pesticide, for example, or plant-based on the market, with an active ingredient that also occurs in plants, for example, then it must also be proven that the product is safe. Because, as you say, it's not just about the presence of a substance, but also the concentration and the way an organism is exposed. I don't want to say that this has to be a problem, I just want to say that this is something you have to keep in mind.

BeeVAX: So finally, could you tell us which aspects of our approach you like? And where are some points where you say, okay, you have to look at it again, if you could find other alternatives.

Dr. Maus: So first of all, I like the idea of a completely different approach with a completely different mechanism. We have two basic problems in the Varroa fight, there are two basic problems of the inherent nature. The first is the specificity. The bees, as insects, and the mites, as spiders, are due to evolutionary relationship in many systems, could be the target for a Varroa fight product. So, you have to find a remedy that controls one and does not harm the other. It's not that easy. We see even with humans how difficult it is to find a malaria remedy because of plasmodium and the human being because of the eukaryotes. How difficult it is with a spider vs. an insect? So that's a new approach. And the other thing is, we have very few Varroa-type active substances on the market, of a synthetic nature, which leads to slight resistances. And the more different mechanisms of action are there, the better you can counteract resistance. To the things that probably have to be thought through well and maybe actually should be reflected again, I don't know if we ever talked to someone from a bee institute, is that the step toward commercial product development is quite large and quite demanding due to numerous regulations, numerous regulatory requirements that have to be met, but also the consideration of how to market it, how to produce it, how to sell it. It's all an incredibly complex thing and it is probably good to work with someone who has experience in the area or can already incorporate infrastructure. And the other point is the cost. So, the key point will be that if you develop a product based on the methodology, that you can offer it at a price that would be attractive for the beekeeper.

BeeVAX: So, you would say from your experience, in terms of production costs, our product would be too expensive for the everyday market at least in the early stages?

Dr. Maus: I don't know that because I don't know what the costs of production or development are. So, you have the development costs and the production costs. And that's something, because I don't have a marketing background and no production background, where I only got what our colleagues told me. But that's a whole different world than the scientific one. And it is really one of the main reasons that we have so many Varroatides. This market does not have a big return on investment like for some other animal species, that would be something to consider. I wouldn't say that this is a showstopper, but that's something you should keep an eye on from the start. [...]

BeeVAX: Thank you very much for the interview, we wish you all the best.

Dr. Maus: Likewise, good luck and success with the further work. I was pleased that we could exchange ideas.

Lobmeyer: Maybe you can tell me what your concern is and what you do.

BeeVAX: Our intention is to get your opinion, especially from an environmental point of view, on our project. Our project is about trying to improve an immune protein of honey bees, so that it can bind better to certain PAMPs of entomopathogenic fungi, and thus make the bees immune to the fungi. And thus find a way to treat the Varroa mite more efficiently, more easily and hopefully more ecologically, by spraying the fungus on the beehive or by doing it differently and kill the Varroa mites with it and the bees survive. Bees have a naturally low immune-defense against fungi because of the low evolutionary pressure. Their reaction is stronger against Gram-positive bacteria. Fungi are not a problem for the bees, because in the stock it is too warm for them. The immune system works theoretically against them, but not really well. We try to improve this natural system. Maybe you can tell us what your impressions are about our approach.

Lobmeyer: So first of all, I'm neither a biologist nor a chemist, nor a Bee expert. As NABU Münster, we are of course interested in the fact that the honey bee is doing well. But our main focus is on wild bees that live solitary and insects in general. And that is of course a completely different topic. There are two main causes of death. One is the insecticides in agriculture and the other is the light pollution. These are our main issues. Before we continue, have you already talked to the beekeepers in Münster?

BeeVAX: We already met beekeepers and talked to different beekeepers. The problem in Germany, we already had an expert interview about, is that most of them are hobby beekeepers. And our product is probably too expensive, especially in the beginning. But for many beekeepers it is the case that they would like a simple solution. Because at the moment a lot of organic acids are being treated, which are not that positive for the environment and which are difficult to use, because you always have to find the right time to have a good yield. And that's why there is a real interest in a more simple and sustainable replacement product. Especially viruses are a big problem. And these viruses are then, for example, transferred from the honey bees to the wild bees and thus also damage the wild bee population in large quantities. But it is of course very difficult for us to intervene in the wild bee population. And now as a human being, of course, I have no access to it. But our adjusting drew are honeybees. We can reach a kind of herd immunity, through the immunization of the honey bees if we manage to control the virus outbreaks in the honey bees, they will be carried less to the wild and protect this population by proxy. This is also a very important aspect of our project.

Lobmeyer: I can't help you in detail.

BeeVAX: The field we work in is synthetic biology, that is, the generation of proteins, organisms or DNA sequences that do not occur in nature, they can be based on nature. In our case, we work with a bee protein itself, that is, not a sample of foreign DNA. What is the position of the NABU when it comes to the application of this method. Or how you would assess the public opinion or the standpoint of the nature conservation authority. Is there a general position that the NABU takes on such GMOs? Or do you do it differently depending on the case?

Lobmeyer: I can't remember hearing that the NABU is positive about genetical interventions. I would assume that there is a general skepticism about gene manipulation at NABU. Maybe you should do some more research on the NABU side, on the government's side, whether they are taking any actions. I can't do that now, I wouldn't know. So, on a local level, it's not an issue at all. It's more like, let's say, I respect your work, I think it's enormous what you do. But it is of course a work, if I see it correctly, a work on the symptom, which is not bad, but the NABU is rather inclined to go to the causes. And when we talk about pathogens, there are of course, besides such virological things, the main causes are, as I said, insecticides and light pollution. We have a very intensive agriculture and so on. We have too little water in the landscape. We have a shortage of species. Not only in the grasslands, but also in our gardens. These are our issues. We have to keep more water in the landscape and so on. We have to get away from our way of agriculture of our whole.

BeeVAX: So, maybe again just as a question of understanding, you would say, especially now, your regional association is not so much concerned with the topic of honey bees, but mainly with wild bees, right?

Lobmeyer: Yes. We want to protect wild bees, of course, because they have a very important factor as a breeder, we can't do without it. That's why we are also very interested in protecting the wild bees. That's for sure.

BeeVAX: Yes, we are aware that the wild bees play a very important role in nature. But the honey bees are closer and easier to influence. Of course, general improvements in economic conditions, renaturalization and so on. Of course, steps that are actually necessary. Of course, this can be implemented very quickly for such a small person.

Lobmeyer: Yes, the insect repression is a key issue. We have somehow lost three quarters of our insect stocks in the last 20 to 25 years. Because of the pesticides and the lack of food for insects, like wild bees.

BeeVAX: Is there any information about the impact of climate change on disease spread? Maybe that diseases from more southern regions migrate further north and meet unprotected populations.

Lobmeyer: Especially among insects, which is probably not the right way to address this, but I know about bird populations, such as the Zootovirus, mainly in the Amsersail. And of course the northern migration of insects. That is clearly noticeable, for example, in different species of butterflies. But as I said, we are a local association and we mainly deal with volunteer work, except for a few exceptions. Scientific work is of course still being done at our NABU nature conservation station in Münsterland. This is the biological station, the network of biological stations of the state of North Rhine-Westphalia, which is responsible for the city of Münster. They have their own extensive website and they receive various orders for cattle from the state or from the federal government or from the EU. I have no idea how far they have gone with the research on insects. You should ask the station. They are voluntary, and the station has 35 voluntary scientists. Biologists, landscapes, zoologists. We are more of the association that represents the members. We have 2,500 members in Münster. We are more there to make us unpopular with the administration and so on. If something goes wrong, raising our finger and complaining is our job. We also participate in various mappings, but the main scientific work is located at the natural resource protection stations. These are not always NABU stations. In Coesfeld, for example, this is not a NABU station. But for Warendorf and Münster, this is the NABU natural resource protection station. Also carried by us and also by the NABU district association Warendorf.

BeeVAX: Alright. Thank you very much for your time.

Lobmeyer: Thank you.

BeeVAX: Maybe do you want to just introduce yourself a bit, what you do, what are your current projects. What are you working on?

Dr. Hopkins: Yeah, sure. I'm an assistant research professor here at Washington State University. I've been working on honey bees for, I don't know, 10 or 12 years or something like that. And I kind of, you know, done a variety of things. I worked initially on like cryopreservation of honeybee semen and doing some breeding efforts, like importation of semen into the US from like native range of honeybees. But the, well you guys probably saw this or assuming that's why you're focusing on Varroa mites, there's like a big pressure from industry and you know worldwide efforts for Varroa control and so a lot of my work in the last eight years or something like that has been more on Varroa control and commercial scale beekeeping management. I do a lot of work with commercial beekeepers and trying to improve management practices in part related to Varroa control. Just overwintering strategies, nutritional supplementing for these folks that are on migratory routes for commercial beekeeping. It can be very difficult to keep bees alive when they're moving the bees all the time on different pollination tracks.

BeeVAX: So you really advise the beekeepers what to do basically or what to improve?

Dr. Hopkins: Yeah, a little bit. Beekeepers are quite innovative and so they try different things. And so, some of them find success in certain strategies or management practices, but they don't have the ability to kind of test that. It's really more like they feel like things are working. And so, then a lot of what I do is really just take their ideas and then put it through a little bit more rigorous testing to see if it's actually improving health of the colonies or improving survivorship or whether it's just that they feel good about what they're doing.

BeeVAX: [description of our project]

Dr. Hopkins: Yeah, I think it's very cool. I mean, I'm assuming you're quite familiar with that work with that Dayland company and the mean priming against foul brood still. I don't know how far along in this process you guys are.

BeeVAX: We are trying, we have like different approaches. We have one approach where we just bind a zymosane, so the pamp of the fungi we're using is zymosane. We're trying to bind that just with the Vitellogenin and then just link a protein to it that binds to the zymosane. That would be like the most straightforward approach, but the construct gets huge after some time and we don't know how well that works in the bees. Then we also are working on directed evolution via yeast surface-display. So, you display a library of mutagenized domains on yeast cells and then sort them via fluorescent activated cell sorting, so you can identify positive variants. With that project, we're pretty far along, so we have the yeast strains presented with the proteins, we have the libraries, and we went sorting two days ago, I think. The cells are currently growing, and we can sequence them later this week. And the last group we have is working on rational design, so looking at the protein sequence, but they have the problem that the protein isn't that well researched, so we don't know the pockets where it binds, we don't know which residues are involved, so it's kind of a collaborative effort between the direct evolution team and the rational design team to figure that out, and hopefully get some ideas there. So, they try to use some binding prediction programs and docking programs, but they don't really work so well with our protein because the binding part is so huge, you know.

Dr. Hopkins: So the idea is that you're using the yeast cells to produce the protein of the entomopathogenic fungus in order to prime the honey bees?

BeeVAX: No, no, the yeast cells are just a tool for developing or for engineering the proteins. So we are trying to engineer the bee protein to better bind the fungus particles. So, the yeast are just presenting the binding domains, and we can mutagenize the binding domains, and then you have this library of different domain variants, and you can screen that library and pick out improved protein variants.

Dr. Hopkins: Okay, gotcha. Yeah, that's very cool.

BeeVAX: So, like, our project probably won't, definitely won't get to the stage that we can try it with bees. That's the point we've gotten to until this point.

Dr. Hopkins: I mean as far as the approach I think yours is much cooler you know you you're familiar with like the literature of using the entomopathogenic fungus to control Varroa mites like Kanga wrote a bunch of papers back in like the 90s or whatever.

BeeVAX: But so, there's still a problem with using this fungi, because they also affect the honeybees or not?

Dr. Hopkins: Well, that's what I was gonna ask you is that, you know, the work that Kanga did, and I can't, so I can't think of his first name or whatever. But he used to work in the US, USDA. And now I think he's in like, Florida or something like that. But so he wrote a few papers. They even, as far as I was told, started trying to develop a commercial strain of that fungus for Varroa control. And then they had difficulties like generating the fungus in the lab. But, you know, the story that we heard, and then we also, you know, for the last four or five years have been selecting a strain of Metarhizium for Varroa control. And in both cases, I mean, to be honest, at least the fungus that we're working with is like not really very deadly to the bees themselves. And we put a lot of that fungus directly in the hive and we did not see a lot of honey bee mortality. And it's interesting that you guys are working on this. I think it's very cool because part of that is a little bit of a mystery in some ways, because it should probably be more pathogenic to honeybees. But so some of the like theories that have been suggested is that the bees have sort of that like static charge on them. You know, they've got all these fine hairs that like the pollen sticks to these hairs and the entomopathenic fungus has to like attach, like touch the actual cuticle of the insect in order to start like generating the tube, you know, the hyphae tube to infest. And so if all those sort of Metarhizium spores are out on the hairs, they're probably not gonna be deadly to the bees in some ways. The biggest problem we have with the entomopathogenic fungus is that it's a soil fungus and so it does not grow well at the temperatures that a hive exists at. A hive is like 34, 36 degrees Celsius when they're raising brood and that soil adapted fungus, in our case which is Metarhizium, doesn't develop well at temperatures really above like 25 degrees Celsius or something like that. Basically, it's not very deadly to the bees. But then in turn it also struggles to be virulent against the mites or stay for very long periods of time. So what we see is a lot of Varroa death within the first two to three days. The Metarhizium is killing Varroa mites for a few days and then it stops killing mites.

BeeVAX: Yeah, we also saw Metarhizium anisopliae, but we also saw something about Beauveria bassiana as an alternative. Hurt the bees definitely a bit more. Like I think a tenth of the colony died when you aplply Beauveria. And that's not really viable for beekeepers to choose that. We chose it because it's more prone to grow under higher temperatures. The exact longevity, we are not quite sure, but maybe we'll need to look into that. The Metarhizium, it would enable, like if we managed to immune prime them better, it would enable us to use more competent fungi in the hives, which maybe might affect the bees more, maybe enable you to use different different fungus strains or maybe like different species, like maybe that would open a door to towards new applications like with Beauveria, for example.

Dr. Hopkins: The other thing I was going to ask, and I'm certainly not trying to change the direction of your project or make it more complicated, but it would be interesting to me to look at if it's possible to, like you said, maybe combine some of these things. They have Beauveria that is maybe more heat-tolerant. I do know people have been working with it, but I don't know that much about it and we haven't. So what we did was more like what you're talking about directed evolution so we grew the Metarhizium in the incubator at higher temperatures and put it with, you know, challenged media growth and then selected strains that grew better at higher temperatures and so we continued to select for Metarhizium that survived at higher temperatures in the incubator but it's pretty slow and it's very labor-intensive and all that. So, you know, is it possible to, you know, modify the Metarhizium or Beauveriaum with, you know, like, some kind of different heat shock proteins or something like to make it more stable at higher temperatures or look at why maybe Beauveria survives at high temperatures and Metarhizium doesn't. And so that way you could have, you know, instead of just doing one side of the story. But can you modify the Metarhizium or the entomopathogenic fungus at the same time there? Because if you make the Metarhizium maybe more pathogenic and you protect the bees at the same time then you get like a more virulent fungus but protecting the bees at the same time so kind of like modifying both organisms in conjunction to achieve like a better Varroa control.

BeeVAX: Yeah, it would be great if both things could work together. One question we always have is like if the fungi work in the hive, is the honey production affected by it? Do you find like decent or like problematic traces of fungus in the hive, which maybe may make it difficult to market or cause allergic reactions or stuff? We know that yeasts are a problem if the honey contains too much water, that yeasts grow in them, but do you have any knowledge if like fungus contamination is a problem?

Dr. Hopkins: We haven't looked for the spores but it's not that it's not an issue I think it's something that should be looked at probably it would certainly be in the spores but like most Varroa control stuff beekeepers don't generally use it while they're making honey anyway. So they apply it in the colony and then stop applying it, and then they put the honey supers on and make honey. Take the honey off and then apply the stuff after they've taken it off.

BeeVAX: Yeah, because that would make it a great option for using it while producing honey. If the fungus isn't a problem, then you could treat a Varroa infestation while still producing honey. And then you would have huge advantages over the other methods, you know?

Dr. Hopkins: It'd be interesting to look at it, and actually fairly simple or straightforward, especially if you have, you know, the, whatever fungus that you're using, have some type of primer ability to look at it, because, you know, there's lots of methods for screening honey for sort of, I think, eDNA. I mean, you could screen it and look for the traces or whatever the presence of the entomopathogenic fungus in samples of honey. It would be pretty straightforward to look at that. We just haven't yet tested whether or not the Metarhizium is showing up in the honey. But it most certainly is. I mean, the honey carries traces of like even Varroa DNA. So they use like, there's a lab in North Dakota that screens, you know, the eDNA of honey and finds, you know, Varroa DNA in the honey and, you know, pollen sample, you know, there's all sorts of pollen grains in the honey and all that stuff. I'm sure it'll show up in there.

BeeVAX: Yeah. One implication of maybe using an improved fungus strain would be that it maybe infects wild bees when the honey bees have contact with wild bee populations. Do you think that could be a problem when you're using like a very aggressive strain?

Dr. Hopkins: I don't. I mean, only because there's a lot of concern about, and this is more related to pollination, and assuming that bees carry the pollen around on the hairs of the bee, and so the concern would be that the bee leaves the hive with this Metarhizium on them and they fly out, and then on a flower they interact with the wild bee and the metarizum kills the thing, the wild bee. I would be less concerned about that, in that they've done a lot of work with looking at whether or not. Like let's say you move a colony from one pollination, like say broccoli, and you move it to go pollinate cauliflower, and they're the same species that cross-pollinate, right? One is just a brassica that's green and one's a brassica that's white. But it's a big problem if you're producing seed, seed broccoli and seed cauliflower. And so they've shown that bees are very, very good at cleaning themselves and they clean themselves before they leave the hive. So, you know, bees' goal is to bring pollen into the hive. Their goal is not to take pollen out of the hive. And so once they come in the hive, they clean themselves and they clean themselves when they leave. So my guess is that bees leaving the hive do not have a lot of this fungal spores on them because they certainly don't have a lot of pollen on them when they leave the hive. And so if they're cleaning the pollen off, I'm assuming that the fungus spores will be cleaned off as well. That would be my hypothesis, but again, it would be fairly easy to test this and you could just capture bees as they're leaving the hive and then, you know, grind them up and look for fungal spores or wash them and then look for fungal spores on the bees. See what the concentration is.

BeeVAX: So we're also thinking about like real-world implementation of our project if it maybe ever gets to a point where you actually use it in a commercial sense. So we have like different approaches. Maybe for the EU we have the problem with GMOs. So we can't just make a GMO bee that produces the improved Vitellogenin. So we have to look for something else. With like, injecting it, it's all like, not really realistic. So, the question would be, do you think that in America there would be an option to introduce bees like that to the market?

Dr. Hopkins: GMO bees? No. I think that yeah, I think I think it's maybe not as much demonized maybe in the US as it is in Europe, but I think honey bees and like flying insects that you can't control so much as like much scarier to the population. GMO bees, I mean, we've heard of this before and there was like really bad publicity. I think it was a lab in Germany that had done some of this work and they'd only done it like in the lab and never produced an actual bee, but they were like injecting DNA into honeybee eggs or something like that and showing the possibility or potential of making GMO bees and it was like, got very, very bad publicity here in the US at the time.

BeeVAX: Oh, I understand. Yeah, because we talked to a beekeeper club here, and they were all pretty convinced by it. They said, like, if it works, give it to us right now.

Dr. Hopkins: I heard the same thing from a beekeeper here, who was saying, you know, I don't really like the idea of genetically modified bees, but if it could solve my Varroa problem, I would take it. I don't think the problem is selling it to beekeepers, but the problem is the public. The public only is like 1% beekeepers, but the 99% of the other people would not understand, I don't think.

BeeVAX: Do you have any questions for us?

Dr. Hopkins: No, but I mean I think it's a very cool project and I would like to... I'd love to hear, you know, some updates or how things go. It'd be great to hear about that.

BeeVAX: Yeah, of course. Great. Thank you very much.

Dr. Hopkins: Yeah, yeah, thanks. Good luck, you guys.