TWiV 1343: Batting averages and stolen bases

0:00This Week in Virology, the podcast about viruses, the kind that make you sick.

0:10From Microbe TV, this is TWIV, This Week in Virology, episode 1343, recorded on July 24th, 2026. I'm Vincent Racaniello, and you're listening to the podcast all about viruses. Joining me today from Ann Arbor, Michigan, Kathy Spindler. Hi, everybody. Here it's 79 degrees and partly sunny, partly cloudy. And that is, I didn't get to calculate it yet, but our air quality is good. It's down in the 42 range. Yes. Not bad. Yes. Here it's 28 and sunny. also joining us from Western Massachusetts, Alan Dove. Good to be here. And it is absolutely gorgeous here, very similar to Kathy's weather, 81 Fahrenheit, 27 C.

1:06And I will also comment on the air quality. We had a day a little while ago that was downright apocalyptic looking. It was just the air was khaki colored from the Canadian wildfires, really bad. And from Austin, Texas, Rich Condit. hi everybody uh 92 degrees um uh sunny but i you know i think there's a kind of a high haze it's a summer here in austin we've had our first couple of hundred degree days uh the uh my car was reading 111 while i was uh riding back from something the other day so uh you know here we go a couple of months of this and three months really Well, July is almost over. Yeah.

1:55And then one more month, and that's basically it. Well, it's Texas. Yeah, it's Texas. You know, I've been known to be having frustrated in October. Yeah, it stays warm for a while. It does. It's just not summer. So after this recording, I'm going to head out to Brooklyn. We're going to have a walk of the Gowanus Canal. a bunch of listeners have responded to our call and we're going to, you know, to go on, this is, is an interesting place. I thought they were cleaning it up, but you can walk across it now. Okay. You can walk along. Along. Oh, right. Okay. Not going to walk across it. We're in it.

2:36Not going to swim in it. No, no, don't recommend that. But there's some very interesting, we did an episode on TWIM long ago and decided we should take a walk and people expressed interest. So I'm going to go out there and walk for an hour. and we have a little sheet that has information about it. You can probably leave your fishing pole at home too. It's nice weather for that, yes. Gowanus. Dixon always used to talk about the Gowanus. If you enjoy these science programs, we'd love your support. Go to micro.tv slash contribute. And the Rosenfeld Lab continues to look for a postdoc and a technician to work on viral pathogenesis and immunobiology.

3:18They work on enteroviruses, they're interested in pathogenesis, immune responses, and developing new vaccine platforms. You can send her an email, rosenfelda at mlhs.org. We do not have a update for ASV because it's starting Monday. And by the time you listen, it'll probably already be started. So have a good time, everyone. I'll be heading out as well. I'm actually going tomorrow to Rochester, Minnesota, to attend a measles mini symposium. That should be fun. And do a pod there as well. And one of my listeners, one of our listeners, is going to pick me up at the airport and drive me to Rochester.

4:06and then he's going to drive me to Minneapolis because he said he wants to talk to me. Cool. How long a drive is it from Minneapolis? So an hour to Rochester. It's an hour to Rochester and then it's an hour from Rochester to Minneapolis. Looking forward to that. Okay, everyone enjoy that meeting. You're going to have a twiv with extra mayo. I have a twiv with extra mayo, yes. And then I'm going to have a Twivit ASV. Always fun. Okay, news. We have two news items. We have the death of Susumu Tonagawa, 86 years old. He got the Nobel Prize for explaining how antibodies are produced and such diversity, right?

5:03We did this paper. It's a PNAS paper a long time ago when we were doing sort of classic papers showing that antibody genes are all for the heavy and light chains and the variable and constant regions. They're all arrayed and they're all different kinds and they're recombined together. And then splicing happens and you get all these different antibody specificities. And that was a problem in immunology. No one could figure out how it worked. And he figured it out. He did the experiments. So very important discovery. At the end of this article, they quote an interview with him where he was asked to define creativity.

5:50And he said, I don't feel I am very creative. I wish I could be more creative. But then he comes up with three simple concepts. One, you have to be curious. Two, you have to have an unfailing urge to address the question. And three, you have to combine knowledge in at least two different fields where people don't necessarily interact. I don't think that's a definition of creativity. Maybe it's how to do good work, but it's not creativity. More of a protocol. Yes, that's what he was, a protocol kind of person, I would guess. Anyway. So I have a distant connection to Sasumu Tonagawa. He did his PhD work at UCSD in the same laboratory where I started my doctoral work, that lab of Masaki Hayashi.

6:36So Susumu worked on PHYX-174. And in Wikipedia, he has an autobiography, but I already found three mistakes in it. So one's a typo. He's got UCDS, but it should be UCSD. And then he says that he received his PhD in molecular biology in 1978 and stayed on as a postdoc there until 1979. But those are off by 10 years, so he must mean 1968 and 1969, because then he moved to the lab of Renato Delbecco at the Salk, where he worked on SV40 to define the transcripts during lytic infection. And since this was pre-restriction enzyme and pre-recommoded DNA age, The information he could obtain was very limited.

7:25But being a member of the best laboratory in the field, I glimpsed the excitement of the cutting edge of scientific research. So anyway, probably, and then, yeah, he goes on and talks about stuff happening in 1971. So it's clearly just a decade mistake. But anyway, that's a little more. So he did have some starts in virology. So that's relevant for this podcast.

7:51All right. So this New York Times article showed up just right before we went on. The title of it is Measles Cases Hit a New Record in the U.S. as Vaccination Rates Wane. This is not necessarily, well, it is news. It's big news. Okay. But something we've been tracking. It's big news, but not surprising. Big news, but not surprising. I'll just read the first couple of paragraphs here because it says the whole story. The United States has now recorded more cases of measles in 2026 than in any other year since the virus was declared eliminated in 2000, a grim sign that the country may be headed toward a new era in which disease regularly sickens thousands every year.

8:42CDC on Friday announced there have been 2 ,318 confirmed measles cases so far this year. That tops last year's record. More measles reported in the United States in the last two years than in all the years from 2000 to 2024. Combined. Combined. Reversing course on measles would require a nationwide push to improve vaccination rates, a prospect that experts said seemed unlikely under the leadership of Health Secretary Robert F. Kennedy. He has halted funding for vaccine hesitancy research, downplayed risks of measles, and spearheaded a hunt for evidence that vaccines are unsafe. I would say leadership is a stretch to describe what he's doing.

9:38Yeah. So we are now seeing, we are now harvesting the fruits of this craziness. Make measles great again. Make measles great again. I like this quote by Jennifer Nudzo, director of the Pandemic Center at Brown University. It's going to mean living in a perpetual state of vulnerability and risk until we get vaccination levels up. the more this goes on, the more likely it is you're near someone with measles. Yeah. And, you know, measles is kind of the canary in the coal mine because it's one of the most, if not the most, contagious infectious, microbial infectious disease that we know of. And so it's sort of the first sign that things are going pear-shaped.

10:27But there will be more to come. Yeah. And I mean, this is overwhelmingly affecting people, kids who were not vaccinated for usually bad reasons. But as these cases become more prevalent, the virus is going to be around people who may have gotten vaccinated a long time ago. Their immunity is waning, who can get reinfected and potentially get serious sequelae, even though, you know, they should have been protected because it shouldn't be this common. A particular vulnerability with measles that we have discussed before is that children, it is recommended that children not be vaccinated until they are one year old because they pick up maternal immunity from their mothers.

11:18And that takes a while, at least six months or so, to wane. And if you vaccinate them too early, their maternal immunity compromises the effectiveness of the vaccine. So you've got to try and pick the sweet spot. You don't always get it. But what it means is that there is a population of children who are potentially vulnerable. And what that means is that if you want to protect them, you need to get the whole population immune to measles so there's no transmission. And we've lost that. The other big group that's at risk, of course, are people who are elderly, whose immunity is waning, or anybody who's got other immune issues who may not be as immune from the virus as they should be.

12:00So, folks, you get to vote on this in November. Yeah. All right, on to the papers. We have first a PNAS, Proceedings of the National Academy of Sciences Research Article. Per-os infectivity factors are essential for brachovirus infection and wasp parasitism. And the first author is Ji Wang. Senior author is Ji Hong Hu. The group is from the Wuhan Institute of Virology in Wuhan, Chinese Academy of Sciences in Beijing, and Wageningen University and Research in the Netherlands. And Rich, you're going to summarize this for us. Yeah, this dude is one of my favorites. I love this. It's so weird. so this is our periodic revisit to the world of what are I don't know if it's with a new taxonomy if they're still called polydna viruses but that's what they were originally for polyDNA having genomes of many many circles of DNA

13:10so yeah and we've done at least two previous episodes that somebody put in here, TWIV 372 and TWIV 443. So we haven't been here for a while. Okay. So in order to describe this, this is a summary, but it's going to take a little bit of time because this is so weird. Because it is convoluted, yes. It's convoluted. Okay. So I will start by talking about polidinoviruses, and then I'll talk about the actual paper. So this is a virus that plays a role in the life cycle of a parasitoid, oh, a virus in air quotes, okay? And you'll see why. It plays a role in the life cycle of a parasitoid wasp. That is, this is a wasp that lays its eggs in a caterpillar of some insect, a moss, for example.

14:08And that provides a nourishing environment for the eggs and larva of the wasp, which then hatch by emerging from the caterpillar and maturing outside the animal. The inspiration for Ridley Scott's Alien. Absolutely. Think Alien. Okay? And I do want to add, we'll be talking about a specific virus here, but this is a phenomenon that appears to be fairly widespread among paracetamide wasps. And what we're going to talk about is that it has arisen independently a couple of times, which is mind-boggling. So the caterpillar has an immune system which will compromise the development of the wasp's larvae.

14:57And the wasp has developed evolutionarily a way to suppress that immune system to the benefit of its offspring. And this involves, during the maturation of the eggs in the oviduct of the wasp, the concurrent appearance of virus-like particles that are filled with multiple circles of DNA. These particles are deposited along with the eggs in the caterpillar. The pieces of DNA, in fact, encode immune suppressive factors. The virus particles, virus-like particles, infect caterpillar cells, and the circular DNAs then integrate into the genome of the caterpillar cells and make these immune suppressing factors, which then suppress the immunity of the caterpillar to the benefit of the eggs.

16:03I say virus-like because, in fact, the DNAs that are packaged are not virus DNAs. They are wasp DNAs that have been specially designated by this system that contain flanking sequences that promote their excision and amplification during the development of the eggs in the oviduct to prepare them for packaging and delivery in these particles. The particles come from the remnants of a virus infection way the hell back in evolution. The virus is probably mostly closely related to a nudivirus, which is a double-stranded DNA insect virus. And some of these nudiviruses can integrate into genomes. And apparently there was an incident multiple times with multiple different related systems eons ago, where subsequent to an integration of a nudivirus genome, everything essentially was lost except for the transcription apparatus, because the virus encodes its own multi-subunit RNA polymerase, and the structural proteins.

17:24And this was co-opted by... So this is in the germline of the wasp, and will be contained in all of the somatic cells of both the male and female wasp, but they're quiescent until the female wasp is developing eggs in the oviduct. And at that time, the virus genes are induced and the RNA polymerase transcribes the structural proteins. And that results in the development of these virus particles, which then encapsulate the amplified WASP DNA for delivery into the caterpillars. I mean, this is insane. It's completely insane. All right. Right. So the WASP basically has co-opted an ancient integration to use some virus genes to make a delivery vehicle for genes of its own that are immunosuppressive into the caterpillar.

18:16Including even the integration down on the other end. The WASP got infected with a virus somewhere back there and stole it for its own parasitic uses. These viruses get called brachoviruses. And we'll use that term, I think, as we discuss this paper. But keep in mind, it's a virus delivery system. It's not really a virus. Okay. Okay. So in this, these viruses. The wasp made a vector. Yes. Yeah. It's a vectored vaccine that the wasp, our immunosuppressant that the wasp created. Yeah. So these viruses are also related to a number of other different insect viruses, notably in this case, baculoviruses, which are insect viruses that are acquired in a, these are real viruses, that are acquired in a different fashion.

19:07In baculoviruses, the caterpillars eat the virus off of the leaves that they're eating. And then the virus infects through the gut of the caterpillar and goes through a reproductive process in the caterpillar. And it's been known for some time that important in that infection is a complex of proteins in the membrane of the virus called HIF. What's that? Ur-os infection factors. Okay? It's a complex of proteins that are required to facilitate the entry of the baculovirus into its worm host. And the authors of this paper noted that the capsids of the Brachovirus and the WASP contain proteins. In fact, in the paper they show it contains a complex that's essentially homologous to this baculovirus complex that's important for entry in an oral infection route.

20:20But, as we've already said, this virus doesn't do oral infection, so they're wondering, what does this complex do? Does it serve the same purpose? What's going on? And basically, they show that this protein complex in the wasp virus is embedded in the membrane of the capsid, that it has, if you force caterpillars to eat this thing, it has no effect on infectivity by oral acquisition, oral infection. But it is required for infection during the normal route of the Brachovirus, that is after injection into the caterpillar and release of the virus into the caterpillar so and per os means by mouth yeah per os means by mouth yep so and actually i will um i will say this is preemptive but you know pox viruses have a very complicated fusion and entry complex in their membrane that's been a kind of a mystery for a while and i wonder i'm tempted to send this paper to you've probably already seen it bernie and um Eugene Koonin and say, okay, guys, is there an homology here?

21:47All right, I'm done. Proceed. Okay. So one of these immunosuppressive genes delivered into the caterpillar by the brachovirus is anchoring. It encodes anchoring, which is immunosuppressive. It regulates NF Kappa B, but that's going to be important. We're going to use that as an assay in this paper. So that's one of the genes that is delivered. But so the paper is mostly about these PIFs, right? They've been found in brachoviruses. They are needed for oral infection in baculoviruses. That's the way these are transmitted, these baculoviruses from one insect to another. And so they're essential for that.

22:29That's why they're called per os infectivity factors. And so oral entry is not part of brachovirus infection, so they want to know what are PIFs doing there in the genome. And they use, as a model in this paper, microplitis mediator brachovirus, MMBV, to study the function of these PIFs. So it's just a specific brachovirus that they're to use for these experiments. Okay. So they have genome sequences of these, their M mediator, right? I like that name. And they have, they can see genes encoding PIFs. PIFs. Zero, one, two, three, four, five, six, and eight. So they purify these viruses from the ovaries of female wasps.

23:22They have a nice picture showing that they're pure. It's a cylindrical capsid inside a membrane envelope, and there's actually a tail-like extension.

23:33They make antibodies against these PIFs, and then they can do western blots and other things, and they show that these are membrane-associated proteins. So essentially, the PIFs of grachoviruses, membrane proteins, they form a complex of about 720 ,000 Daltons. Big. It's big. Yeah. Big. But PIF 5-1 and 5-2 don't seem to be involved in this complex, even though they're there. So they do some mass spec to look at the precise components of these PIF complexes. and they find the PIF proteins. But they also find many WASP proteins in these PIF complexes. And they make some antibodies against these proteins.

24:29And only antibody against a protein, which begins with LOC, and I'm not going to tell you the rest because we're going to rename it in a second. Only the antibody against LOC recognizes the PIF complex in Western blots. This is a 52 kilodalton protein, so they call it P52. Yeah. Which is a famous protein in mammalian cells. There's P53. P53, sorry, P53. Never mind. What is P52 that I'm thinking of? It's this. Okay, so it's not a... No, there have been other 52K proteins, and I had the feeling P53 might have been P52 at first. Okay. That doesn't really matter. This is insects. This is P52. So P52 is transcribed exclusively in female wasps within the ovaries, and the expression coincides with the developmental period of the brachovirus in the wasps.

25:27So this is a wasp gene encoding P52. Now, remember in Rich's summary, brachos are derived from ancestral nudiviruses, right? There's no sequence homology between P52 and any nudivirus genes. They use alpha folds to predict its structure, right? So that's a nice program that you can use. You got an amino acid sequence. It has a huge database of structures it can predict. So this protein has a typical beta propeller domain with seven blades surrounding a central cavity, and each blade contains a four-stranded anti-parallel beta sheet. Now, to many of you, that may sound like meaningless, but a beta propeller is a known structural domain.

26:19Also, they mentioned something about a Velcro thing. Yes. But I couldn't see that in the picture. A Velcro mode to close the circle between the first and the last blades, right? Because you've got these blades, you've got to close it up. And again, they don't find this domain in any nudiviral or baculoviral proteins. And also, there are introns in this gene for p52. Unlike viral genes, the virus has no introns. And so all this suggests that it is a host gene instead of a virus-derived gene. They also find p52 in a couple of other wasp species. So it's a wasp gene that's forming part of this complex with the piffs.

27:06That's quite interesting. So then we do know that the brachovirus structural protein genes are driven by viral RNA polymerase, right, in the WASP. And so they want to know if p52 is dependent on the same polymerase. So they knock down a subunit of the RNA polymerase, and they find that PIF0 and PIF3 are downregulated. But p52 is not altered by knocking out the RNA polymerase. So again, it's derived from, that's the viral RNA polymerase that we're talking about here. p52 is derived from the WASP, more evidence that is derived from the host rather than the virus. Okay. So then they decide to knock down individual PIFs with double, You can introduce double-stranded RNA into these cells and knock down individual genes.

28:04So they microinject double-stranded RNA into newly formed WASP pupae. And basically knockdown of any subunit gene, any PIF, results in disappearance of all the others. So they say the whole thing comes apart when you just take one away. So it's not stable unless it has all the components. So the complex relies on every subunit to maintain integrity. That must have been interesting to see that result. Yeah. Take everyone away. Each one one at a time. One at a time, and it breaks it every time. So what's the function of these PIFs in these brachoviruses? They're going to silence PIF, and then they're going to infect SF9 cells.

28:53Spadoptera frugiperida. What are they? Butterfly cells, I think? Army worm. Army worms. And the insects that they're studying here, the specific parasitoid system that they're studying here, this wasp parasitizes also an army worm and another moth, both noctoid moths. What is the other one? Oh, it's cotton bowl worm. I should know that one. So, yeah. Yeah, the SF9 cells are in that same general vicinity. So they knock down the genes again with double-stranded RNA, and they look at the bricovirus morphology and virus yield. And basically, there's no effect on the morphology and yield. The control is a different double-stranded RNA, and they all look the same.

29:51Okay, so then they infect SF9 cells with different depleted brachoviruses, and they quantify virus infectivity. They use the virus-encoded anchor in mRNA, right, which we said before is a marker for the immunosuppression. So they find that anchor in transcripts in the knockdowns are 10 to 25 times lower than in the control group. So viral infection basically is hindered when you knock down the PIFs. So the virus particles that you make are okay, but infection is messed up. And interesting, this PIF-5-1 and PIF-5-2, there's no effect on transcription, which is a measure of infectivity. So they think that they're just paralogs that compensate for each other's function.

30:40Okay, so that is, it's involved in infection, right? Now, they want to know if the piffs could mediate oral infection of lepidopterin hosts. So, bacchuviruses, remember, they depend on oral infection to spread. But the brachos get injected into the insect body. There's no role for feeding there. So, it's wasp oviposition. So, could brachovirus piffs mediate oral infection? And I guess what they're getting at here is, do these PIFs retain their original function? Yeah, that's right. So they ask if you can measure infection with their model virus by anchoring expression, and they inject them into the hemoseal of the hosts, and they can see anchoring expression.

31:33So that's how they're going to measure infection, by measuring anchoring, okay? And then they do an oral infection assay. They feed native or heat inactivated virus to two different larvae. They quantify anchorin 24 hours later and no infection. So these brachoviruses are not orally infectious. You can feed it to them. Nothing happens. All right. So what do you need a piff for then? And then if it does something, it's going to have the wrong name. Doing it for us, yeah. That's the life. So what's the role then of PIFs? So then they look at systemic infection. They inject the virus and they got PIF-depleted brachoviruses into the hemolymph.

32:24And then they measure anchorin transcripts. And so when you do that, anchoring is much lower in the knockouts or the knockdowns with double-stranded RNA for PIF-03, 52, 5-3, 5-1, blah, blah, blah, compared to the control group. So the PIF complex, and as well PIF 5-1 and 5-2, which seem to be separate from the complex, are critical for systemic infection, for brachovirus systemic infection. So they're not involved in feeding because the virus doesn't infect when you feed it. But if you inject the virus, it reproduces, and these PIFs are needed. So they're needed for infection.

33:15How about immune suppression? So the brachovirus infection of the host larvae immunosuppresses them, as we have said before, and that allows the development of wasp eggs, as Rich said. So they ask whether knocking down the PIF genes would affect this wasp parasitism. So they have a parasitism assay, which they use, they do with PIF knockdown wasps, okay? And so they inject these, the wasps are injecting them into the larvae. And if you use, again, the knockdown for these piffs, most of them, the wasps grow faster than the control group. All right. So knockdown of, you know, piff 0, 3, 52, et cetera.

34:04They grow faster than in the control group. the average parasitism rate for the control is above 90%, and if you knock down these PIFs, it drops to 0 % to 8%. So you can't establish very efficient parasitism, in other words, infection in the wasp, if you don't have these PIFs. So they conclude that the PIF complex and PIF-1-2, which are separate, are essential for parasitism of these hosts by mediator, M-mediator. So they're important, but not for feeding. So BIF isn't the right name, but that's okay. But exactly what they do, we don't know. They're per ovipositor infection factors. Oh, yeah, you could modify it to be that.

34:59POFs. POFs. But, well, they would be POIFs if it's per os. So per ovipositor would have the same problem. So some of the take, those are the experiments, straightforward manuscript, take-home messages. So this PIF complex in the brachovirus is a WASP. It's quite a WASP protein, P52. It has a beta propeller domain. And that kind of domain usually mediates protein-protein interactions, which is what these PIFs are. So maybe it's a scaffold for assembly of the PIFs. But they say that this is an example of the domestication of a host gene by a virus. So, you know, several things here. First of all, the interesting thing is that you have a protein complex made by a gaggle of genes that in two different instances, like the baculovirus and the brachovirus, is purposed for catalyzing entry, but the roots of entry are quite different.

36:08And they don't cross-react. It's not like it'll enter anything. They're specific for the virus. So it's interesting that over time you take something for a certain purpose and adapt it to a modified purpose.

36:27I was disappointed that they didn't do knockdowns of P52, the WASP component of this, to see what effect it has on the complex. And I don't think they even did the experiment to see if knockdowns of P52 compromised the infection. So maybe that's coming down the road. I'm always fascinated by methods in particular. in a paper like this. And if you, the method section in this basically, essentially just refers you to a supplementary document that is wonderfully detailed. And it describes how this whole life cycle and all of the insect components for the life cycle are reared in the laboratory and how that all goes on.

37:19It's fun to imagine this. And then just a couple of things. One of the things that they did in this to isolate this complex was to isolate a bunch of virus particles, brachovirus particles, and then solubilize the membrane proteins and show that this thing was in the membrane proteins. So to do that, about 200 female wasps were collected and dissected for ovary isolation. And they harvest the ovaries and do a couple of things to isolate these by sucrose gradients centrifugation. But I'm imagining dissecting wasps for ovary isolation. These are not big creatures for harvesting this stuff. They wind up with a half a mil of stuff, I think.

38:10And I was interested in the knockdown. So they make double-stranded RNA and they get this. They inject pupae and let them mature to adults and then assess the adults. So they've got double-stranded RNA. Wasp cocoons formed within 24 hours. that is 24 hours after they form from the host caterpillar, were selected for microinjection. Cocoon is placed on a glass slide using double-sided tape. A small hole was made with tweezers in the abdominal region of the cocoon. Approximately 400 nanoliters, that's 0.4 microliters, of... Oh, dang, I lost it here. Sorry. Yeah, 400 nanoliters of double-stranded RNA were injected into wasp pupae using a Nanojet 3 injector at an injection rate of 200 nanoliters per second.

39:23Adult wasps emerged four to five days post-injection, and female wasp ovaries were dissected and assessed individually. So that knockdown is done in the pupae and is good enough to the genes stay knocked down through the rest of the maturation of the adult wasp. And so they can look at what the effect on the wasp ovaries. But what tedious work. You've got to really want the answer to do this. Yep. Now, there's a sentence in the discussion that I don't understand. Maybe you guys do. They say, this study demonstrates that in brachoviruses, pifs are dispensable for oral infection, but essential for systemic infection.

40:15But there's no oral infection, so how can they be dispensable? Well, they are dispensable if you orally infect with these viruses. No, no, they didn't get an oral infection. Exactly. Oh, the dispensable. Okay. Dispensable is the wrong word. Yeah. Okay. So there's no oral infection, but they're essential for, you know, because later they say we show that there's no oral infection process. So they can't be dispensable for a process that doesn't exist. I just thought Kathy might have picked that up too. I did have questions about systemic versus oral because they just kind of seemed to sneak that in and I didn't really get it.

40:58But I think I get it now. But the technique that I thought Rich was going to comment on was this blue native page, which is a gentle laboratory method to separate intact protein complexes. instead of using SDS that denatures the complexes, it uses Kumasi Brilliant Blue dye, and that keeps them folded and bound together. So that's how they were able to look at these 720 ,000 Dalton complexes and so forth.

41:33Always interesting stuff. I love this system. I really respect and admire the people who work on it because it's absolutely fascinating. Yeah. Yes, I always like these wasp papers. So as soon as I see them, we pick them. Okay. And if you're somebody who, you know, is always looking for the applications, I mean, these are, in this particular system, these are two major, major agricultural pests. The one of them attacks rice and corn and the other one attacks cotton and some other crops. And these brachoviruses and this wasp parasitoidism occurs in many other agricultural pests as well. Our second paper is in Science Advances.

42:17Periodic shifts in viral load increase risk of hendrovirus spillover from teropus bats. First author is Tamika Lunn, and senior authors are Raina Plowright and Allison Peel. And the group is from everywhere. Okay, so University of Georgia in Athens, Wildlife Health Ecology Research Organization, San Diego, University of Antwerp, Belgium, Montana State University, Bozeman, Texas Tech in Lubbock, the NIH campus in Hamilton, Montana, Mississippi State University, Starkville, Cornell, UCLA, Griffith University in Nathan, Queensland, University of New South Wales in Sydney, Penn State in State College, University of Cambridge, UK, University of Waterloo, Ontario, C-SYRO and Canberra, and University of Sydney.

43:09So, Kathy, you have a summary of sorts, right? Of sorts, yeah. So, I couldn't figure out where to start, and so I'm going to maybe come around and repeat myself at the end, but they're going to be looking at two measures. One is viral load, and the other is prevalence. And I struggled with figuring out what they meant by each of those. But eventually, I found the viral load definition in the materials and methods. And then prevalence, I kind of just got by osmosis. So as you could guess from the title, they use hendrovirus, which is a bat-borne negative strand RNA virus that causes severe disease and death in horses and humans in Eastern Australia.

43:52The virus was first detected in 1994. And the primary host is terropus bats, in particular black flying foxes, although they mention gray-headed flying foxes and little red flying foxes in their study. So they're going to challenge this current paradigm that higher prevalence in viral shedding in reservoir hosts, that is in bats, is a lone driver of spillover from wildlife to humans. So prevalence, okay. And they find, however, the authors find that there's a temporal shift in the distribution of virus loads as the seasons and years change. And if you can include this viral load information over time with prevalence, that will give you a better correlation with known spillovers and therefore might allow you to better predict future spillovers.

44:52overs. So the significance is that if they could do this, it would give you, as I said, better information about virus, infected versus infectious states, virus distribution across landscapes, and the doses that are available for recipient hosts. Okay, and so I'm going to put in two things here that I think Vincent may want to comment on later. One is the statement that it is widely accepted that individuals with higher viral loads are more likely to be infectious, and they gave three references. So in the course of doing this experiment, let's see, they collected pooled urine samples from under bat roosts from five sites over three years, all in Australia.

45:45So that doesn't, At first, I thought all these places that these people are from is going to explain their sample collection. But no, they're all collections from Australia. Okay. Anyway, and it's all along the coast near Brisbane in eastern Australia. Okay. So, yeah. Okay. So what they conclude, we're going to go through this, but that previously reported low-prevalence shedding, so that you can measure this, but this low-prevalence shedding doesn't align with spillover risk. And so this is going to quote them directly, that prolonged low-intensity shedding of Henrovirus outside of the winter months, which is detected only by using the conventional threshold.

46:36And I'll insert here that the conventional threshold to them means a CT value of 40, which means doing many cycles of PCR to detect something that's present in very small amounts. So this prolonged low-intensity shedding may reflect either prolonged shedding of non-infectious RNA or viral loads that are too low to overcome dose barriers to spillover infection. So they're saying that if you have these long stretches of time with low levels of Hendra virus, that doesn't seem to correlate with spillovers. but they do monitor the levels using this high CT value because it's such a deadly virus that you want to be sure that you always detect it if it's there so

47:39what do they mean by viral prevalence the proportion of positive pooled urine samples. Okay. And then what do they mean by viral loads? And this is again where I think Vincent might want to comment because he's commented in the past about the use of the term viral load. But what they say is, we use the term viral load throughout to refer to proxies, in other words, genome copies and CT values. Although we note that the term, quote, viral load is used beyond the historical virological definition, which only uses TCID50 or platforming units to describe viral load. Okay. I thought that line might be in there for us.

48:29Yeah, it might have been. So, okay. So, I'll just interject here. This is why I had a little bit of problem, because they measured viral prevalence as the proportion of positive pooled urine samples. So how did they determine whether something was positive? Well, they did it by doing PCR and calculating CT values. So it seemed a little circular to me. But anyway, at the end, they even say they use this phrase in other words. So I was able to search for it and find it again. In other words, while low viral loads may contribute to transmission and maintenance of hendrovirus within the natural bat reservoir host, they appear to pose minimal risk for cross-species transmission, i.e.

49:23spillover. Peak prevalence periods associated with spillover events, N equals 5, had a higher proportion of samples with high viral loads than periods without spillover. So if you had a period of time, like in the winter, where there was very high prevalence and they had a high viral load, then those would correlate with spillover. So that's my summary, but as I said, of sorts. So I had the same sort of difficulties with prevalence and load, but you have helped me out significantly here. But I want to circle back and see if we can summarize this. Is it fair to say then that prevalence is basically how frequently, that is, what fraction of samples that you take at a given time in a cave contain virus using the really sensitive test?

50:26Is it there at all, even using a very sensitive test? It's plus, minus, yes, no. Plus, minus, yes, no. But using a very sensitive test. And then load is have a closer look and tell me in any given sample kind of how much there is. Yes. By doing a more quantitative assay. Fair enough? Yes. Yes. In fact, you hit on something that I had written in my margin and forgot when I was writing out my summary. that maybe prevalence, in quotes, is a plus-minus measure and viral load is quantitative. Yes. In case they use CT, but they'd be better if they used black assay and TCID50. Well, right, but an important, okay, 6 ,151 unique pooled urine samples from 152 sessions.

51:16No, they didn't use TCID50. They wanted something they could semi-automate. Yeah, of course.

51:50and load is inferring load from data that don't necessarily just give you, you know, a TCID 50. So there's a lot of statistics that has to go on here. Right. So that's basically the background for this paper, that in the past people have used prevalence, plus or minus. So you get a bat, is it infected or not? There's no quantitation, right? And for obvious reasons, you can imagine that that's not good enough, right? So you could be positive, yet hardly any infectious RNA, right? So it's not really useful to include that. So that's what they're trying to do here is to see if we quantify it, at least by CT.

52:32It's not ideal. And they do admit at the end it would be better to have a different quantification. But this is just to see if there's a pattern, how they're going to try to do this. And they use Hendra as a model to do this because they have a lot of data on Hendra. Hendra, you know, this has been going on since 1994 in Australia, in the northeastern part of the country mainly. and there have been 68 spillover events, 110 confirmed or suspected cases in horses, seven confirmed human cases. And this is a high-priority pathogen of WHO because it's quite serious. I have a miscellaneous question.

53:23Am I correct that horses and humans are dead-end hosts for this, that they do not transmit it from one animal to another? So really we're— The humans get it from grooming the horses. Oh, okay. Fine. But humans don't transmit it. So the horses eat contaminated fruit or whatever that the bats have, or they contact urine from bats. I don't remember the spillover. And then people who are grooming the horses get infected as well because there's no contact between the bats and the people. All right. So there's a figure showing. They say despite the alignment of flying foxes and horses across most of eastern Australia, most of eastern Australia, spillovers have only been detected in coastal New South Wales and Queensland.

54:14So coastal regions. And most of these occur in the winter. And they say maybe that's due to increased prevalence in bats in the winter. and previous work also suggests periods of low off-season prevalence in roosts from subtropical Australia and consistent low prevalence shedding from roosts in tropical Australia. Why these don't translate to spillover is not known. We also don't know why cumulative low-level shedding over a long time is the same risk as high-level shedding over a short time. And so they're saying maybe if we do viral load, we can figure this out to a certain extent. The timing also has to do with eucalyptus and food availability.

55:06So, yeah, in winter, there's not very many species of eucalyptus producing nectar. And this coincides with the higher levels of hendrovirus shedding. and that may explain the stronger predictive power of models that use the number of spillovers rather than occurrence. Okay. So I wanted to just tell you how they collect these urines. All right. So these bats go to known roosts. Okay. They go back to the same one every night. They're in trees and you put plastic sheets on the ground. I remember this because when I was in Australia, they brought me to a bat roost. And you could see the plastic sheets on the ground.

55:49You could smell the urine. It's incredible. It smells like a urinal in a bathroom. It's just amazing. And they put the sheets there before sunrise. And then the bats come back at sunrise. They're out all night. And they collect. And the bats are in the trees. And they're peeing. And And they pool the urine on one sheet, about 20 droplets to a—it gives you about 2 ml. And, you know, if there's poop on it, they don't use it because they don't want that contamination. But I remember Lin-Fu Wang used to tell me from one—he used to pipette up a drop of urine from the sheet. He said, I can isolate multiple herpes viruses from one drop of urine from these bats.

56:35It's really amazing. They were gray-shouldered bats. They had actually established a roost right next to the laboratory. Very convenient. So they could go right out and study it down in Geelong. So that's how they collect these samples. And as has already been said, they have they have they have 6 ,100 pooled urine samples from 152 sessions 2017 through 2020. and 4 ,657 of these, they recorded a black flying fox, one or more, so that they want that. Those are the ones they're going to use for the black flying foxes for this. It's the primary reservoir of the Hendra virus. And so you see cycle threshold 40 as a threshold, right?

57:24If you have to, you don't go above 40 because there's just nothing there if you go above 40. It's just noise. So that's the threshold, CT40. 361 were positive for Hendra, 8%. And the mean viral load is 22 ,000 genome copies per mil. So as Kathy mentioned, people now use viral load for PCR, whereas historically it's been infectious. Daniel Griffin always makes a point of saying genome copies, which is what they say here, genome copies. It shouldn't be viral load. It should be genome load. But it's there, and we're not going to change it. We're just going to be complaining. They do have in their discussion a couple of sentences that say, to further explore the relationship between viral load proxies and infectiousness, further studies could formally test a serial dilution relationship between viral detection, genome copy numbers, and virus infectiousness using TCID or a black assay.

58:24Yeah. That would be a good thing to do. And I don't know if it's going to be constant all the time, right? That's the thing. You find a relationship in one study, how do you know? But you have to do it at some point. And it could be variable from lab to lab. That's right. Anyway, so what they find is seasonal and interannual patterns in shedding prevalence. Okay, so that's the original thing. And then they use their calculations of normalized pool's preference, and they find that with the high CT values, the excretion typically peaks in the winter, which is June to August. The low prevalence shedding is apparent through the rest of the year.

59:03And then using lower CT thresholds, they find pooled prevalence is minimal outside of winter and large peaks were not observed every year. So then they want to test whether viral load and prevalence are associated. And what they find is that the divergence between observed and permuted normalized prevalence curves identifies periods when the observed data contain more or fewer positive than expected under a random distribution of viral loads. Right. So it took me a while to get my head around this permutation analysis procedure. They're going to sort of shuffle the data between the controls and the experiments to compare when we shuffle the data, do we get the same distribution as when the data were unshuffled as a test to see if this is a real correlation.

59:59That's my short, possibly a little, you know, wiggly answer to what they did. All right, so this permutation analysis shows strong support for a CT distribution shift toward disproportionately lower values, higher value loads. So the low CT, higher viral loads during high prevalence periods. And higher values, lower viral loads during low prevalence periods. So there is a correlation between the two. And across the CT threshold, high quantiles coincide with high prevalence period and low quantiles coincide with low prevalence period.

1:00:46They also align cases of spillover, right? We know when spillovers happened, when there was an outbreak of Hendra and horses and people. So they align those, and they find that the cases align in these normalized pools, the prevalence estimated using the lower CT threshold value. When this time series is subdivided based on season and spillover, normalized prevalence was highest for peak seasons in which a spillover event occurred. And then for peak season with spillover events, normalized prevalence tended higher for lower CT value. So in other words, there's a correlation, a rough correlation between spillovers and the CT values.

1:01:34And that's an inverse because the lower CT value correlates with the higher viral or genome load. So basically what they have found, they have found periodic shifts in the load shed by the bats. And it skews toward higher loads during high prevalence periods. All right. And this supports the idea that the spillover risk increases when both prevalence and individual viral loads are evaluated. You got to do them together. And so they say the conventional detection at CT40 may not be the best threshold for predicting spillover. And then they go on to talk about what Kathy has already said. You know, this PCR test was developed to say, is there Henry here or not?

1:02:21Because we want to know who's got it and where it is and so forth. It's not meant to assess spillover risk, but people have been trying to do that. And they say, you know, modeling tools often assume that RNA detection equates to infectiousness, but not necessarily. It really depends, right, on the CT.

1:02:45So what they say is that prolonged low-intensity shedding of the virus outside of winter months may reflect either prolonged shedding of non-infectious RNA or viral loads too low to overcome dose barriers because we don't see an association with spillover for this prolonged low-intensity shedding. And maybe it's, of course, it's probably important for maintenance of the virus in bats. Yeah, absolutely. Right, but not for spillover. The bats need to pass it around, and they don't have to have too much virus to do that. But you know, there's 8 % of the bats. It's not a lot, I suppose. It's high enough so that I'm wearing some PPE when I go into that cave.

1:03:21Yes, for sure. Then let's talk a little bit about food because Kathy mentioned this before. So temporal availability of the eucalyptus blossom, right? That's the preferred food of these boxes. Low food availability and utilization of alternate foods may lead to reduced immunocompetence and increased viral shedding. So if they don't eat the right food, we all know that. You got to eat the right stuff to have a good immune response, right? The bats are no different. And in the winter, there aren't enough species making nectar. And that period of scarcity coincides with high levels of virus shedding.

1:04:05So this is the climate-driven food shortages, basically, are probably driving some of this increased levels of virus. And then, yes, we should do dilutions and compare CT values with TCID50. But that could be, as we just said before, that might not help all that much. So basically, they say we challenge the paradigm that higher prevalence of shedding in reservoir bats is a lone driver of spillover from wildlife. You have to also account for viral loads. and that's probably going to be influential in future similar studies, right? So, fun fact that I learned when I was in Australia, I was talking with someone and they were talking about koalas and what they eat and they said something about gum trees and I said, they eat only eucalyptus and they said, same thing.

1:05:13So gum trees, the majority of trees called gum trees are eucalyptus. And so the kookaburra sits in the old gum tree. It's actually, they're sitting in probably a eucalyptus. All eucalypts are considered gum trees, botanically speaking. But not all gum trees are part of the eucalyptus genus. So anyway, that was my fun fact about eucalyptus and gum trees. The California coast is a very similar biome to Australia. So I grew up on a street lined with eucalyptus trees. They're all over Marin County. I love those trees. They smell great. They're beautiful trees. They're messy because the bark sheds all over the place.

1:06:00But they're very cool trees. And not to be confused with the gum gum tree, which is from One Piece. just for the listeners who might know that the the second time i've told this story today and that is that when i was there i was on a tour where he took us to a place where we could see koalas in the wild in the trees and he bent over and he picked up some koala poo and he snapped it open and he held it up to our noses and it smells like eucalyptus it smells beautiful It's the most amazing poo. So there you go. And eucalyptus, if I'm not mistaken, is toxic to a lot of other creatures. Is that right?

1:06:46I think that's true, yeah. The koalas have a way around that.

1:06:53All right, let's do some email. Kathy, can you take the first one? If I can find where my document is. There we are. Okay. Charles writes, hello, Twivers. Cool and wet today in Burlington, North Carolina, 70F21C with rain. We need the rain. Dr. Derek Lowe has another excellent post titled, The Assault on Science Funding Continues. It is well worth reading. And he gives a link to the science article. Another music pick from my generation, not the Who. Cat Stevens just had a birthday, 7-21-1948. One of my first albums was his Tea for the Tillerman from 1970. Later, Charles. P.S., you did ask for more letters.

1:07:45It's fine, Charles. Thank you very much. And I remember hearing Tea for the Tillerman all over my freshman dorm. Absolutely. Yeah. So this is a really nice article by Derek Lowe. I just want to read one line, a couple of lines. The Trump administration hates academic science funding. Full stop. They hate where the money goes. They hate who it goes to. They want to keep all the money for themselves to hand out to favored cronies who can help get them elected and steer more money and more power back into their hands. That's it. They are not putting it in those words, but instead talking about gold standard science and reproducibility crisis and funding bold inquiry for transformational breakthroughs.

1:08:31This is all BS. It's important to realize that. Shameless, hand-waving BS delivered in a how-dare-you-think-otherwise manner is the defining style of the entire administration. You can see it in every appointee, in every part of the executive branch, and in the pronouncements of all of his supporters. I realize this is a simple heuristic to apply and a very unambiguous and unflattering one, but I find it to be all too useful and all too accurate. Give it a try. Gaslighting. Yes. Gaslighting. Gold standard science, as if we haven't been doing that for the last 50 years. How do they know what gold standard science is?

1:09:16No. It's not. No. These are words they're throwing around. This is, yeah. Yeah. They're throwing words around. It's all BS.

1:09:27Well, there's no such thing as honesty or truth in any of this. It's just you say whatever is convenient to promote your agenda. That's right. There's no respect for the truth at all. there rich can you take the next one brian writes hello twiv during the question session with john die someone i think from africa asked three questions of which two were addressed the third was what about vector migrations has this been researched rodents bats etc do not stay put we assume that virus outbreaks happen when mutations overcome immunity but it may be also a function of a slowly spreading vector, which takes generations to move from one region to another.

1:10:16When vectors cross, naive hosts, they may be far from a previous outbreak. And this is from Brian. I am not familiar with the John Dye interview. What's this about? Well, someone said basically, well, first of all, he says, we assume virus outbreaks occur when mutations overcome. No, not necessarily. Yeah. I mean, you have a new virus in a bat or whatever, and it goes into a person who has no immunity has never seen that virus before. So there's no mutation involved. It's just a spillover. Right. And so, yeah, I mean, migrating animals could play a role, but we have no idea what the, what the res, as, as, um, Tom Kysak said a week ago, was it just, was it last week?

1:11:07Two weeks. We don't know what the vector is. So, you know, we can't really speculate on migrations because we don't – if it's a bat, there are different species of bats. So we have to figure that out. Well, speaking of vector, we just did a paper, okay, that says that the virus load in a given vector can vary. Yes. Right? And that contributes to whether or not there's spillover. That's right. Vector I traditionally think of as being an arthropod, but the reservoir. Yeah, exactly. Yeah, but the other thing is, you know, there are many examples of people looking for and finding migrations or animal movements that are directly connected to spillovers.

1:11:49West Nile virus pops to mind. You know, in drought conditions, you get birds in places they aren't normally, that kind of thing. so two comments um alan you have to ask your daughter uh what vectors are that is what yeah it's a very different thing in her world than in mine and um john dye was on that special episode of twiv that vincent did from the asm meeting about ebola virus and andes virus okay good and do you know what you get when you cross a mosquito with a mountain climber uh-oh nothing you can't mix a vector and a scalar oh oh it's a math thing that's a math thing yes uh alan how old is your daughter now she's 20 wow holy crap yeah how does that happen she's she's she's going to be going into her junior year at umass oh i know it's you think you think you're you're mortified yeah you remember little harper yeah who used to yeah you know come on the podcast and then and she's going to college next year.

1:12:58Yeah. Wow. I was going to say something else, but I've lost it. So I hope that answers your question. And I keep – by the way, I do keep telling Sophie that she should kind of pursue, if she goes into research, something in the biological realm. So maybe we could have her on TWIV. Sure. No, but she would not want to do it. No. Come on, Dad. I don't want to do that. Okay. Who's next? Alan, please. Sure. Anthony writes, provides a link, and this is from New York City Parks and Rec. Is that it? Oh, Pigeon Rescue Central. Sorry. So this is an independent organization, I think. Quoting, have you ever seen a pigeon that has missing toes, a foot, or even a missing leg?

1:13:48The chances are that the digits and or foot were lost from string being wrapped around them. Wow. And thank you, Anthony. That is a longstanding mystery. I lived on Staten Island for a couple of years, and on the ferry, there were pigeons. And, well, on Staten Island at our bird feeder, it was all pigeons. And they were very frequently missing bits of feet. And I always thought it was from something like landing on a hot tar roof or a piece of air conditioning or something. But this is interesting to know. Well, Angela, last week, I think her pick was pigeons. And I asked her, why are they missing feet and toes?

1:14:23So Anthony is responding to that. Okay. And Anthony continues with the pigeons, by the way, are fascinating. Darwin formulated the theory of evolution based on his experience with pigeons and artificial selection. His observations of the Galapagos finches were featured to show that the theory had universal relevance through natural selection. Gives another link to the preserved samples. Darwin kept his own fancy pigeons at Downhouse, Kent, and many were later prepared as skeletons. His collection of preserved pigeons eventually comprised around 120 skeletons and skins. Fancy pigeons were especially important in helping Darwin understand how features could be selected and inherited, and how one species could produce many different forms.

1:15:06As a result, pigeons are discussed at length in the first chapter of The Origin of Species. Yes, I remember that. Anthony continues, by the way, doves, pigeons represent, in case anybody was wondering why I have an interest in pigeons, doves slash pigeons represent a milestone in human history. the transition from ice age hunters of large mammals to farmers and herders. The Noah story, with its rising waters and collection of animals, alludes to the melting glaciers and the trial and error of pioneering animal husbandry. Some, like the dove that returned home, were amenable to domestication. Others, for example, crows were not.

1:15:43Cool. All right. Gary writes, the junior senator from Delaware, Lisa Blunt Rochester posted a news article on her personal Facebook about the first confirmed measles case in Delaware in over 10 years. Sadly, almost half of the emoji responses was the laughing emoji. Three hundred and forty out of six hundred and ninety eight. I fear my state is headed to an infected summer. Laughing emojis? I think probably a lot of those were people who feel that, you know, well, I got my vaccine, so this is a funny thing for those anti-vaxxers. I think you're being charitable. I'm being a little charitable, perhaps.

1:16:29I haven't been on Facebook in a very long time. But yeah. And even if that was what they were doing, that also comes from ignorance. Because as we said at the top of the show, nobody's safe with this much measles around. Kathy.

1:17:15That's mine, born almost exactly five years after you. The differences from the boomers are significant. On a recent TWIV, 1341, I think, you mentioned not having gone to Woodstock. For me, Woodstock wasn't even on my radar as a possibility. The two biggest differences between Jonesers and boomers, in my opinion, are the economic outlook when we finished our educations and the Vietnam War. I bought my first house in 1983 when mortgage rates were just over 13%, down from around 16 % a year or two earlier. Five years before that, they had been about 9%. And I was never threatened by the Vietnam War.

1:17:54I've never registered for the draft, and I'm not breaking any law by not having done so. I did have friends whose older brothers drew draft numbers. I never really felt like a boomer, except for enjoying the movement. Music. Here's a good explanation of Generation Jones, and he gives a website and a music pick from my youth, The Guests Who Share the Land, Wikipedia on the album, and he gives the Wikipedia link to about Share the Land. Thanks, Charles. A side note here on draft registration, I'm not entirely sure that you weren't breaking the law by skipping it. I had to register, and I'm Generation X.

1:18:35And in fact, I remember a question ticking a box on a grant application in graduate school confirming that I had, in fact, registered for the draft. Gen X is 60 what? 66? I guess 60. I thought it was from right after the boomers, but I guess it's 66 up to sometime in the 70s or early 80s, and then we get into millennials. Gen X everybody forgets. When do the boomers end? Oh, God. I keep wondering when the boomers are going to end. Oh, no, no, I'm sorry. I'm sorry. No, no, no. That shouldn't have been out loud. I guess according to this, it would be 1953. But I thought that 54. No, it was 64. Yes.

1:19:21Yeah. Or 64, right. Yeah. I mean, we are all boomers except... Except me. You. Yes. So you're a Jones? No, no, I'm a Gen X. You were born in 66? Don't you forget about me. Yeah, so, I mean, we're okay. We're boomers. No, not all boomers are the problem. There are a few that I would point to who are particularly prominent problems. Yes. That's probably true in every generation. It is true in every generation. It's just that the boomers overwhelmingly hold the levers of power right now, still. I don't have any power, Alan. Yeah, but if you look at the people who do, In fact, I would say the three of us here don't really have any power.

1:20:11No, none of us actually have a whole lot of power. We're just doing our things, which, oh well. But I think in every generation you can find people that are problematic. Oh, of course, of course. When was the first boomer? What year? That would have been right after World War II, right? Yeah, 45, 46. So 45 to 54, or no, 60-something, whatever. Okay. um who's next rich can you do the next and last one john writes i expect that you may regret getting into the music of people of a certain age because you may get an avalanche of mail this is okay but here's mine with references to science whenever possible on understanding lyrics i may already mention that i once paid a visit to a lab populated with all indian technicians They were listening to a country music station.

1:21:06I noted on that, and they replied, Oh, yes, it's the only music that we can understand the words to. It's pretty clear. ZZ Top. One evening at a restaurant in Slovenia while at a meeting there in 2010, a waiter saw me and exclaimed, Mr. ZZ Top. Well, I do have a long beard. the next evening at an outdoor concert i ran into him again and he insisted on buying me a beer because of that billy gibbons was also a big fan of a group that you've probably never heard of but if you listen to them you may understand why mando and uh mando and mando and the chili peppers was an early conjuncto group from San Antonio discovered in 1957 by a recording exec who was unexpectedly forced down by a January snowstorm in Denver where they were playing.

1:22:08Their only album, 1957, features the band swarmed around a 1953 Packard limo in front of a Colorado potato field. We're really getting into the weeds here. it looks like a typical album cover from the late 60s but this was a dozen years earlier and a very early LP after listening to it I made a copy for a friend who replied why have we never heard of these guys with the exception of one forgettably novelty track the whole album is an excellent collection of covers in their style you can find most of them on YouTube. I particularly like Cherry Pie, Harbor Lights, Congo Mambo, and South of the Border.

1:22:53Billy Gibbons spent many years searching for the leader of the group, Armando Almendarez, finally finding him in Chicago where he had been playing for many years with Eddie the Chief Clearwater, who was a black blues man who performed on occasion in an Indian headdress. Such were the days. Allman Brothers. You may know that after their demise of their keyboardist, Chuck Lavelle, started playing with the Stones. After their demise, their keyboardist, Chuck Lavelle, started playing with the Stones. But wait, there's more. He's also a Georgia tree farmer and author and a big fan of the American Chestnut Foundation, as is Dolly Parton, and their science-based mission to restore the tree to its native range.

1:23:44Buffalo Springfield. Now we're talking. This is one of my favorites. Buffalo Springfield. You may know that it was because Neil Young was driving a 1953 Pontiac hearse that Stephen Stills recognized him, who he had been looking for, in traffic in L.A., which led to the formation of Buffalo Springfield, which of course eventually devolved into Crosby, Stills, Nash, and Young. Later, the Stills-Young release, Long May You Run, is essentially a memorial to Young's first hearse, a 1948 Buick, something that I only learned years later. Young also, I think, still has a 47 Lincoln Continental named Abraham, But he may be more famous these days for his 59 Lincoln Continental convertible that he converted to electric.

1:24:39If nothing comes to mind visually for a 59 Lincoln Continental, they were enormous, bearing no resemblance to the 52-7 Lincolns that were arguably a continuum, or the 55-7 Continentals, or the Continentals that followed beginning in 1961. The thought of converting one of those to electric seems madness, but Neil Young proved that it could be done, so he deserves a techno salute. And there's a YouTube link here where you can learn more about that. Air quality stats. At one point, we were right in the middle of a swath from Canada here in Greater Braddock, and I believe we hit 323 Thursday evening.

1:25:24It was interesting to follow the band of hay shifting from tracking to the southeast, then just straight east as friends on the fringes of either side in central Massachusetts and northeast North Carolina got into the path and then backed out. It could be worse and may well get worse before the season ends, but for now, the orange clown train wants to sanction the Canadians basically for being victims of the very conditions their train seeks to prolong. At least we weren't closely downwind of a burning Russian oil refinery or a shadow fleet ship. We had microparticulates. I understand that those things generate macroparticulates.

1:26:08Best regards, John. Holy cow, John. What a wrap. Thank you. So if you guys remember, after the Tom Kysak, we were riffing on music, right? Right. And so Tom wrote me and he said, just wanted to say how enjoyable participating in the TWIV last Friday was. I enjoyed the discussion that followed my interview, particularly the discussion of boomer music. Please extend my appreciation to your co-hosts. So he liked the boomer. He obviously went and listened to it because he left and then we discussed that. And by the way, I'm going to throw in a bit of Gen X music link to this. I actually went running to search when you mentioned the Mando and the Chili Peppers.

1:27:05Look up the song Cherry Pie, and I had to see if that was the same as the warrant hit Cherry Pie, but it's not. Okay. Look up Dylan Memorial, Eric Clapton, Don't Think Twice on YouTube. And you will see an Eric Clapton rendition. At a memorial concert, 30-year, actually, anniversary concert for Bob Dylan. This was a long time ago. Eric Clapton rendition of Don't Think Twice. Who knew Don't Think Twice is hard-ass blues? It's amazing. Oh, Kathy's pasted in all the generations. Well, so I went to first, I said Wikipedia generations, and then I got the AI overview. So then I went to Wikipedia, and then you have to disambiguate and find and scroll down to list of social generations, and it gives them for the Western world and other worlds as well.

1:28:06But there's dates given in this simplified version, the lost generation, the greatest generation, the silent generation, baby boomers, generation X, millennials, which are also known as generation Y, and generation Z, and then generation alpha. When did all this silliness happen? you know a long time ago after the baby boomers it was the boomers fault like so many things it was the boomers fault yeah i had this conversation with my family the other day the baby boom was a real thing yep yeah right of course because there were a bunch of kids born after the war but all this other stuff is crazy yeah well it's like naming you know all those eras the mesozoic paleozoic you know Jurassic can you keep any of them straight except when you're in a museum that has some pictures for you those actually have defined inflection points as do bronze age iron age I mean you can point to specific things and sort of so do these you know and there are certainly cultural differences between the generations yeah welcome to TWIV folks here we are Okay, let's do some picks of the week.

1:29:25Kathy, what do you have for us? So I have two links. The first is, I think I've picked this before, but you can sign up for something called Science Advisor. It's a daily newsletter, and it's free. You don't have to be a member of AAAS or anything. And it just puts stuff in your inbox that is really fun to read about. And so the pick that I picked for today is from that. And the gist of it is that there's a bioarchive preprint that describes the fact that goats probably do get brain damage from headbutting. So it's long been assumed that they must be protected because of the shape of their skull and strong neck muscles.

1:30:10but this study in a preprint goes and looks at it and there was precedent for brains of dead musk oxen having an abnormal version of the protein tau which is that protein associated with neurogenic diseases like Alzheimer's but this team only has a sample size of three They had three six-month-old male goats, and they named them Alvin, Simon, and Theo, as in Theodore, from the chipmunks, for those of you who might not know. Got it. Okay. If you're not a boomer, maybe you don't know that. Anyway, so they looked at these three six-month-old male goats and recorded them for six months. They had a whopping total of 227 headbutts a day.

1:31:06Um, each month they sampled, uh, blood saliva and brain fluid to look for markers of brain injury. Um, and they even, uh, at the end of the study took them for, uh, positive emission tomography MRI. So a PET scan basically. They were pet goats. Yes. We put them on the human scanner. It caused quite a bit of ruckus when we went down the hallway and there's like two horns poking out of the gurney. so anyway um it appeared that they had accumulated mild uh brain injury and when they uh euthanized them and looked at their brains they found the abnormal tau proteins wow and so possibility is that maybe goats would make a nice model but then there has to be a naysayer uh an animal behavior and welfare scientist in hong kong who says we shouldn't jump the gun the history of biomedical research is replete with findings from animal models that did not translate to humans.

1:32:07I think we need a high threshold before embarking on this. So. Nice lie. Yeah. And so I haven't gotten into this bioarchive preprint yet, but what was the control group? Yeah, right. Goats with helmets. Well, you could have male goats who you isolate in pens by themselves. I do think that I saw that there were... Or you could strap pillows on their heads, which would make for some good photos. Yeah. Isolation might introduce other confounding factors. It might. These abnormal tau proteins, at least, they found were not present in two younger goats. Okay. Okay. So that was the only control that I saw.

1:32:52I didn't look at the bio-i-cide article yet. I'll wait until it's peer-reviewed. Yeah. Yeah. Rich, what do you have for us? So this is just a fun article that I found in the Times. I think I was drawn to it because of the artist's illustration at the top. It's titled, This Star Just Ate a Planet, and It's Not Done Yet. So this is about a star that has some crazy letter number name that's 1 ,300 light years away, which is not all that far. It's about 30 % larger than our sun. And there is evidence that this thing, that one of its planets fell into it and was destroyed. And it describes the circumstances that might have caused that and how it might happen again to other planets in its realm.

1:33:51By the way, stars eating planets is a normal phenomenon for the end of the star life cycle. They eventually blow up and consume many of the planets in their orbit. Several billion years from now, we're likely to at least get hot, if not consumed. Mercury and Venus are probably toast. At any rate, the way you observe this is by the star having, I don't know if it's a spot or generally distributed, high concentration of lithium. Apparently, if you eat a planet, the planet's got a lot of lithium in it and it leaves a residue. But I think more interesting in this case, the reason is that there is another body orbiting this star that is a brown dwarf that is an object that is somewhere between kind of a planet and a star.

1:34:56It would like to be a star, but it doesn't have enough mass or hydrogen to really burn. And this system has a brown dwarf orbiting this star really close. And get this, it orbits this star once a week. I mean, that's really moving. Plus, it's a short orbit as well. And apparently, because of its gravity, it influences the orbits of all the other planets around and causes them, at least this one, theoretically, to spiral into the star, and others are at risk as well. So it was just a fun read. There is a link, there are several links to the articles that talk about this, but there's one that is a link specifically to the brown dwarf that accesses a NASA site that describes this system and the orbiting brown dwarf.

1:35:57It doesn't really talk about the demise of the planet. But it's a very cool NASA site that you can look at the particulars of this particular system and the orbiting ground dwarf. So there you go. Nice. Cool. Space. Alan, what do you have for us? I have a game. So this is a board game called Schisto and Ladders. And it's themed around schistosomiasis. It's the old Snakes and Ladders game. You probably played it as a kid. Shoots and Ladders. Shoots and Ladders. Right. Well, it was... Okay, so this is a very, very old game concept. Snakes and Ladders was the original... Is that right? Yes. And it goes back to ancient India as the original game, which was to teach moral lessons of Hinduism.

1:36:51and then it was reprinted or stolen probably by the British who made it into snakes and ladders and then it was republished by Milton Bradley as shoots and ladders. The concept's all the same. We've probably all played it. But the innovation here is that it's themed around schistosomiasis, which is a parasitic infection. I'm sure if you listen to TWIP, you're familiar with this. You get this worm by contact with contaminated water. It's not, you don't have to drink the water. You could just go dangle your feet in it. And in tropical countries where it's endemic, it is a very common, very serious health threat.

1:37:29So this is an effort to teach kids about schistosomiasis and how to avoid it. And they adapted chutes and ladders, snakes and ladders to do this. And it's a very simple board. And they show somebody shaking the dice in a little pill bottle. So you can make it with available materials in a lot of places. And I'm also linking to a paper about it where they tested this as an intervention to help control schistosomiasis in school children. And they found that it actually is fairly effective for being a really cheap, easy thing to implement. Cool. Interesting. Yeah. It's nice. my pick is a it's an article in National Geographic how female dolphins know which males to avoid this is great there's also a PNAS the original article in PNAS I'll link to that as well but basically this is work done in Shark Bay Australia they found that female Indo-Pacific dolphins are more likely to steer clear of males with a history of consorting females.

1:38:41So this is where alliances of males work together to keep a female close for weeks so they can mate. And during this, the males can bite them, they can hit them, they can charge them, chase them, and produce threat vocalizations that appear to intimidate them. So the males have these social abilities to do this. But the females remember them. And they say, you got to keep away from that guy and that guy over there and so forth. And the lady, Leila Saig, who is a dolphin communication scientist, said females are using knowledge of individuals. That, to me, is super interesting. These are bottlenose dolphins.

1:39:23They're known for their signature whistles and calls that function like— Some of those calls are saying, that guy's an a-hole. Yeah, exactly. Stay away from Charlie there. He's really a loser. So there's also, I'll put a link to the article as well. We have listener picks from David. The video link below caught my attention as one that would be especially interesting. Given your studio location, along with the micro TV content, the practical engineering YouTube channel videos are one I look forward to viewing upon release. This one gives an overview of the complexities of the New York City infrastructure that lies underground.

1:40:01I hope you will enjoy it as much as I did. Be sure to check out the past content for plenty of other fascinating videos and provides a link for that. Thanks for all you do to advance science education and awareness. And then Ken writes, apropos the frequent defense of curiosity-driven research, I recommend this listener pick. Provides a link to a 1939 essay in Harper's by the founder and head of the Institute for Advanced Study. I've seen this before. The Usefulness of Useless Knowledge by Abraham Flexner. It's a classic, yes. Thank you very much, Ken. So I found one follow-up to mention. Yes, Rich, eucalyptus is poisonous to humans and pets.

1:40:54Deluded products like lozenges are okay, but pure eucalyptus oil, either oily or even through your skin, can cause severe seizures and nervous system depression. So don't try this at home.

1:41:08That's TWIV 1343. You can find show notes at microbe.tv slash TWIV. If you have questions or comments or picks, we want to hear from you folks. TWIV at microbe.tv. If you like our work, we'd love your support. Microbe.tv slash contribute. Kathy Spindler is Professor Emerita at the University of Michigan in Ann Arbor. Thank you, Kathy. Thank you. This is a lot of fun. Rich Condit is an emeritus professor, University of Florida, Gainesville. He's currently in Austin, Texas. Thank you, Rich. Sure enough. Always a good time. Alan Dove is at alandove.com and turbidplack.com. Thank you, Alan. Thanks.

1:41:48It's always a pleasure. I'm Vincent Racaniello. You can find me at microbe.tv. I'd like to thank the American Society for Virology and the American Society for Microbiology for their support of TWIV, Ronald Jenkins for the music, and Jolene Ramsey for the timestamps. You've been listening to This Week in Virology. Thanks for joining us. We'll be back next week. Another TWIV is viral.