that was a pretty good intro i hope i can live up to it so uh like dr laflin said i am daniel whitefield uh the six years that i spent here were some of the best of my life uh and i'm really excited to be back and i'm excited so many of you all came more nervous but excited so uh like you said i work in the lab of dr zach edelman at a m in the entomology department um i had zero experience with entomology before i started so i've been learning a lot and it's been a lot of fun so uh yeah i guess i'll uh start explaining how we're engineering mosquitoes to suck less so all right so why why do mosquitoes suck uh well it's not for food is not for food uh because they uh mostly feed on nectar so males and females both feed on nectar um it's so that they can make eggs so this is one of the egg cups it's about like size of a shot glass which none of y'all know what a shot glass is it's a small glass about this big uh and so so we put some paper in there and then some water in the bottom so it gets the paper wet so they'll lay on a wet surface just above the water line and so that means only females suck blood um and it's because blood has a lot of protein in it and so they use the extra protein from the blood to actually make the to generate the eggs uh all right so how do mosquitoes suck uh so they have fairly complex mouth parts uh and so the first part is the labium which is this gray the thicker gray part in the back and so you can see that back here it's flexible it kind of bends back and then the more stiff part of the proboscis actually goes into the skin so that's the labium back here and then the labella are these pads or lips here that make contact with the skin and then they have these a m colored maxilla and so these are they have little saw teeth right there and so those are so sharp that you don't even feel them cutting and so that's that's what saws into the skin and then their mandibles uh hold the skin and other flesh and tissue apart so that the other two parts can go in and so that's the hypopharynx which is this bright green color and so that has the salivary gl duct and so that injects the saliva which has anti-clotting factors it has i think it's it's got like a painkiller so you don't really feel it until they fly away and different things but it also sits on top of this trough that's the labrum and so it seals the top of this uh and makes that a tube and so that's the actual blood feeding tube so that's that's what uh so that's what stabs you that's that's what uh they suck the blood out with uh so that's that's the business end um so how bad do mosquitoes suck uh spoiler alert pretty bad uh so malaria is uh transmitted by mosquitoes and it killed 627 000 people just in 2020 that was down a little bit because we've gotten better at mitigation strategies and decreasing mosquito populations and increasing protection against mosquitoes and malaria and different mosquito-borne diseases but it's back up since then because coven 19 has interfered with some of the mitigation strategies and so the numbers for 2021 and probably 2022 will be higher than that um before some of the mitigation strategies got started i think it was up uh it could have been as high as like two million a year um and uh some of the some of the symptoms of malaria are splenomegaly and hepatomegaly so the spleen and the liver will swell and so you can see a child here um with a swell a swollen spleen and swollen liver so it's a pretty horrible disease if untreated and it's actually it's estimated that five percent of all the people that have died in history have died from malaria so mosquitoes are probably not even probably i think i can be pretty sure mosquitoes are the most dangerous animal on the planet they kill a lot of people every year the next disease is dengue or dengue virus 40 000 people a year there's a pretty characteristic rash from dengue virus some of the other viruses that i'm about to talk about have very similar rashes associated so yellow fever virus called yellow fever because it affects the liver and so it can cause jaundice and so eyes turn yellow skin turns yellow and so that that's been a problem for a long time zika virus you may remember from several years ago when i was still here actually and so in pregnant women infected with zika virus that can cause microcephaly in babies and so you can see that this baby the cranium and the brain hasn't had the cranium hasn't fully developed and so the brain hasn't had enough room to fully develop and then we also hear a lot about west nile virus and chikungunya virus and then something not in humans but i think everybody probably has a heart for i didn't plan that to be a pun um but uh dyrofilariasis um which is heartworms and dogs um and so something that any any pet owners are really concerned with that's also spread by mosquitoes so just some quick facts about mosquitoes there's 3 600 different species but they're they're mostly selective feeders and so uh specific mosquito species will usually feed on specific uh target species and so only a handful of the 3600 actually are selective to humans and so there there's mosquitoes for uh it's usually specific species of mosquitoes will be targeted to snakes frogs birds dogs uh all that different things so there's only a a small fraction of the 3 600 that are targeted towards humans and then only a even smaller fraction of that number actually spread disease in humans and so those are mosquitoes from the genera 80s anopheles and culex and so 80 spreads most of the viruses so dengue yellow fever all of the viruses are spread by 80s anopheles spreads the four different subtypes of malaria and then culex spreads falaria which is a round worm disease and then certain bacterial infections also and so this is aedes aegypti which is the particular species of mosquito that we study most commonly in the lab we have i think we have some monopolies and maybe some coolex but we don't do a whole lot with those 80s are just much easier to work with in the lab and so we we generally study 80s and we'll generalize that to the other species and so i try this isn't necessarily to scale but i tried to eyeball it to where the relative size between the female up here and the male is about what the relative size difference is because the females are larger but other than the size you can tell females and males apart from their antenna so the females they have some hair coming off of the antenna but it's fairly sparse and so when you when you look at the female antenna they usually just look like uh just one kind of prong coming out but the males the hair is much more pronounced and so they kind of look like feathers and then these organs here just above the proboscis are much smaller in females and then here you can see that they come out and sort of makes their proboscis look like a trident so you can tell that fairly easily and then the size of the abdomen obviously females have to fit eggs and blood meals and so they're much larger and the males are uh much more anemic and then something that's unique to 80s species of mosquito is they kick their back legs up when they land like this and so they use those as extra sensory organs and so they can they can sense air movement and some other stuff and so it's sort of like they turn their back legs into an extra pair of antennae um and they also don't need those um though their back legs aren't that necessary so when we do genotyping we just rip their back legs off yeah uh so this is the global range of 80s aegypti and so you can pretty you can see that they're pretty much everywhere that is wet enough and uh warm enough for mosquitoes uh so you know not many in this area where there's no water uh not many uh in siberia where it's too cold but pretty much everywhere else uh you're gonna get 80s aegypti all right so the life cycle of 80's agyptai so it starts with a blood meal um and mosquitoes will lay 40 to 50 eggs per blood meal so think about all the times that you've been bitten by mosquito and multiply that by 40 to 50 and that's how many mosquitoes are made of your blood and so so 80s in particular will lay their eggs just above the water line in whatever container they can find and so when it rains the water level rises and when the eggs get wet then they'll start hatching out pretty soon after that and so that's why you see a huge population explosion of mosquitoes after it rains and then there's four different larval stages so they'll do a molt and get bigger each time and then they'll turn into pupa oh i have pictures too so this is a line of embryos that i lined up myself uh i've gotten better at it so they're straighter now but this one's a little crooked and then just just so you can see how big these are that's my finger for scale so we do that under the microscope and then these are larva these are l4s so these are about a centimeter maybe a little bit longer than a centimeter um and so we put those on petri dishes and look at them under the microscope and then pupa so uh i think it's in my other talk tomorrow so if you want to know why their eyes are different color come tomorrow but it's pretty cool all right so how do we make them suck less that's what we're all here for uh so uh so there's different prevention initiatives and so these are these are ways to control or to sort of organize uh the different control strategies and one of them is just public sanitation infrastructure so a lot of the places that are the most affected by mosquito-borne diseases have poor public sanitation so this is an open sewer and so just designing uh just health sanita sanitation infrastructure having close sewers having just just clean water supply etc helps with mosquito-borne diseases and then public health education so just educating people on how mosquitoes reproduce what their life cycle is how they spread disease etc uh that that can really help and then community engagement is usually organizing communities to go uh check for stagnant water reservoirs and just dump uh spare tires i tried to get the nastiest one that i could find uh and then uh but not just spare tires obviously like tarps that can hold water or you know buckets or any any sort of thing that can hold water just getting the community involved to check around their house check around their neighborhood and just remove those places where mosquitoes can spawn and then surveillance and monitoring is usually more like scientists doing genetic testing to see what species of mosquitoes are entering certain areas keeping an eye on mosquito-borne diseases prevalence and uh just uh that so that's what surveillance is is uh looking at the types of mosquitoes looking at the types of diseases and keeping track of how those numbers are changing and so all of these uh are just ways to organize people so that we can go through the control strategies for mosquitoes so there's four different main uh categories of control strategies so the first is mechanical control and so that's things that mechanically keep mosquitoes away or keep them from reproducing so mosquito nets to protect people while they sleep removing stagnant water building design so this one other than just getting a picture of a door or a window i didn't have a great picture for it but it's it's basically just having doors that seal well when they're closed so that mosquitoes can't fly in in a gap underneath them windows that seal also in a lot of places where mosquito-borne diseases are really rampant they'll just have corrugated steel roofs and a lot of times there's gaps in between the top of the wall and where the corrugated steel roof is so just sealing those holes so it's it's mainly just designing buildings and making sure that they're difficult for mosquitoes to get in to bite people while they're asleep or while they're you know sitting and listening to some crazy guy give a lecture and so then there's chemical control so obviously that's insecticides and those can come in a couple different forms so you can put it in the water and it'll kill larva as they're spawning and then adulticides is pretty self-explanatory but the scale there's there's several different scales that that can happen at so that can just be at the scale of the household so you can go to the store by insecticide it could be industrial scale where it's a guy in like a hazmat suit with a fogger just spraying huge areas with insecticide it could be at the governmental level with literally just trucks with foggers spraying insecticide through a whole neighborhood i wouldn't recommend ddt um this is an old picture uh with it's not used very much anymore uh but uh and then the next two uh so there there's different categories within or there's there's different end goals i guess for the next two control mechanisms one is population suppression and so that's of it to reduce the population so that that's generally stuff that kills the mosquitoes keeps them from reproducing and so those are usually things that are lethal to the offspring and then there's population replacement and so that changes the population usually with the goal of making them less pathogenic all right so the third the third control mechanism is biological control and so uh so you can just add a fish in the genus gambusia because they feed on larva so that would be population suppression because they're eating the larva keeping them from becoming adults then there's wolbachia bacteria which is a intercellular parasite so it's bacteria that gets not just in bodies but inside the individual cells and so this can actually be population suppression or population replacement and so if you uh infect just males with wobachia and then release only uh male infected wolbachia or male mosquitoes infected with wolbachia then when they mate with females there is a cytoplasm mismatch between the sperm and the egg and so when the mosquitoes lay eggs they just don't hatch and so that's essentially making the males infertile and then you can also do population replacement and so you can deliver genes with the wolbachia and so the wabaki will produce whatever protein or compound you want in the mosquitoes and then that can you can target that to have to make them less pathogenic so it could kill the pathogens it could kill the mosquito if they get infected with the pathogens there's various different strategies but so that one you can release females or well females are more useful so you can release females and so when they mate with wild males then they infect everybody and so then it just changes the population to a population that's resistant to whatever pathogen you're targeting uh and then there's bacillus phtharingians israelinsis bacteria which acts as a biological pesticide and so this would be a population suppression mechanism and so what that is is it's a bacteria that infects the protists and other things that the mosquito larvae eat so that all the dots inside this are this bacteria and so then these things are that and so the larva eat their natural food but their their food has been contaminated with this bacteria and then what this does is it will cause perforations in their uh digestive system and kill the larvae and so it's basically just it's a biological pesticide um and then there's genetic control and so you hear a lot about uh this is something that's actually being employed um so sterile male release and so they usually just breed huge amounts of mosquitoes in a lab and then irradiate the males to sterilize them and then release them and so when they mate with females then the eggs that the females lay are just infertile and so they don't hatch and then there's also a genetic modification so we're getting uh closer into what i actually do in the lab and so the and similar to wolbachia this can be either population suppression or replacement um the problem with normal genetic modification is you release genetic or one of the problems is you release genetically modified mosquitoes and then just because of normal inheritance that genetically modified gene is diluted once they start mating with wild mosquitoes and so our next strategy to prevent that is gene drive and so that causes the modified gene to push through the population at a higher rate than it would naturally um so if that sounds scary you're not alone because what if what if it goes wrong uh so uh what our lab specifically focuses on is making gene drive quote unquote biodegradable and so what that means is that the modified gene deletes itself over time and so what you can do is field test it because laboratory testing can only get you so far it's small populations and so you can't it's it's much more difficult to see exactly how it's going to propagate through the population and so you need to do field testing but if you're testing it it may not be ready and so you want it you want the field testing to be temporary so if something's not quite where you want it to be if there's off target effects or you know something bad happens you want that to be temporary and if it's out in the wild you can't just go around with a butterfly knight and catch them all and so uh so making it biodegradable is a way of making the gene drive delete itself and be temporary over time so that in this is completely without our interference we just design it to do this automatically and so it makes uh the genetic modifications uh delete themselves so that they don't stick around forever so let's talk about the science um so uh so i know a lot of you probably have taken dr laughlin's class taken other people's class but uh i'm gonna start basic and then get actually fairly technical with this talk so we're going to start slow and build from there so so what is dna it's a molecule that stores the information needed to make proteins and so uh so we learn uh i mean most people just intuitively know that they need protein in their diet but that's sort of any protein uh works for you know dietary reasons for nutrition but if you zoom in to specific proteins each different type of protein performs a specific task so there's a bunch of different proteins doing a bunch of different things and what i have focused on with my research is uh the proteins that do dna repair because a third of your entire proteome is dedicated to protecting the information that uh is used to make your entire proteome and so a third of all the genes that you have code for dna repair proteins um all right so getting into dna damage so there's many different causes some of them you can prevent some of them you're just stuck with so the natural ones that you're just stuck with are from when dna is copied you can just have replication errors and so you can't avoid that that's just going to happen but that's why a third of your genome is for repairing there's also just natural metabolism generates some harmful different chemical species mostly different types of reactive oxygen that'll just oxidize whatever they come into contact with i can't get away from that either because you can either deal with this or not eat and die so and then there's radiation which is one of them more avoidable certain types of radiation like just sunlight is hard to stay away from but not drinking from the water fountain at chernobyl you can avoid that and then there's chemicals so these are more avoidable so don't smoke and then and you know there's various different carcinogens that you can avoid there seems to be more in california but that's another story also chemotherapeutics so the way that most chemotherapy works and this is more related to what i did in grad school but the way that most chemotherapy works is it damages dna hopefully more often or more severely in cancer cells than in normal cells but most chemotherapeutics are targeted at either reducing your cancer cells ability to repair dna or they directly cause dna damage so there's different types and they fall into a few main categories so there's breaks so you have single strand breaks that just break uh through one of the backbones so dna is double-stranded so it just breaks through one of the backbones double-stranded dna as you might have guessed breaks through both strands and so you have a full disconnect and so these are just you know free independent moving if you have a double strand break so that's the most uh damaging no it's the most uh deleterious kind of dna damage and then some of the other ones are cross links and so you can permanently bind the two strands together which you want them to be able to unzip so you don't want them permanently bound together and then there's also you can bind base pairs within the strand and so that just that's keeps proteins that need to move down the dna strand from moving down the dna strand or bind binding to and then there's modification so that's things like from oxidation from reactive oxygen uh there's other things that it's just like chemical boondoggles that just get added to the dna that's useless and it it prevents the proteins that read the dna from being able to do their job uh and so what i focus on what i focus on in grad school what i'm focusing on now is uh double strand breaks and then there's several different ways that double strand breaks can be repaired and so get into some of the more technical stuff so there's multiple pathways and and so you want you want multiple safety nets to make sure that the damage gets repaired and so having multiple pathways gives the cells multiple tools to use to be able to repair the damage and so some of these particularly the double strand break repair pathways can be used by scientists that do genetic engineering and so some of the pathways will focus on because they directly have to do with the research that i've been doing so one's called single strand annealing or ssa and then the other is called homologous recombination which is hr so single strand annealing so you have this double-stranded dna and then these these purple boxes are just sequences in the dna that are the same as each other so so this is the exact same sequence as this so the way the single strand annealing works is if some damage happens in between these two repeats and causes a double strand break then there's proteins that will bind and then chew up one half of each of these strands and so you get the single strand so the single strand annealing so it chews it up gets the single strand and then these repeats since they're complementary they'll stick together and so the the repeats will stick together and then you'll have these single strand pieces just kind of sticking up out and just in the nucleoplasm and then they will be clipped off by specific proteins and so then you'll have that and then that's a fairly easy thing for other proteins to come in and stitch back together so you've gotten rid of the double strand break so that the dna is repaired but as you can see there's a fairly significant deletion so you're losing information and then the second one is homologous recombination so this is the most difficult for the cell to actually do because very specific conditions have to be met so you have to have two copies of dna so this will only happen during specific times within the cell cycle because you need two copies because it uses one as the template to repair the other so if you have some damage event causes a double strand break then proteins will come in bring bring the extra copy into close contact and there will be enzymes that there will be proteins that read the template copy and just copy that over to fix whatever got deleted or damaged here and so then once it's repaired then you have fully intact there's no deletions there's no insertions there's no mutations so it's the most accurate but it's also the most difficult to do and then so this is not dna repair this is one of the more recent genetic engineering tools and so this technology has only been around for about a decade and it's only been super popular since i was started grad school so uh so crispr cast nine you can think of it as a set of scissors with a mat and so the scissors are a cas9 protein which cuts dna and then the mat is sgrna and the sg is a single guide so single stranded guide rna because it guide because it's a map and then rna is a very similar molecule to dna and so it has a sequence on the single guide rna that will bind to a very specific sequence in the dna and so you can you can tailor this to target a very specific place within the dna so if you want to cut something or insert something at a particular place in the genome then you just give the cast 9 the right map and then it'll go and it'll bind to that particular sequence and cut the the dna and so if you just have these two things together then you'll just generate a double strand break at that just after that sequence but if you get it some sort of cargo gene if you're trying to do genetic engineering then when it cuts it'll insert that cargo gene and some of the cells natural dna repair mechanisms will make sure that that gets fully incorporated and is just part of the genome now all right so getting into gene drive so normal inheritance so if we just made genetically modified mosquitoes and release them and didn't have any sort of gene drive mechanism so normal inheritance you have this genetically modified male and we usually use males just because they're it's much easier for them to spread their genetic material they can mate with three to five females before they need a break and then it's usually three four days later they can make with three or three to five more females and so you make sure that uh their uh genome from their dad or their their set of genes from their dad and the set of genes from their mom is both this both has this genetic modification that you've given them so when they mate with wild females that have just the normal wild type uh set from their dead in the normal wild type set from their mom then they'll make a hybrid uh it'll be male and female but i just use male uh because it's easier for the for the figure so uh so you'll have hybrid mosquitoes um and so obviously the uh from their dead it'll be genetically modified and then from their mom it'll be wild type and so then they'll mate with more wild-type mosquitoes and so you'll only have 50 percent of those will be hybrid and then 50 of the grandchildren will be wild type and so you can see uh the next generation it'll only be 25 percent and then 12 and a half percent etc and so you can see that this is washing itself out of the population it's not really pushing through the population with gene drive you start with the same thing so when it mates with a wild female and you get the same hybrid but with the gene drive what it does is it'll copy the genetically modified copy of the gene from the dad over the mom's copy that's wild type and so you'll go from just a single copy out of the two to a double copy from both essentially from both parents and so then when it uh mates instead of 50 hybrid 50 wild type they're all hybrid and then the same thing will happen so you have both copies again and so that just keeps pushing through the population and so that that's just gene drive uh in general and so uh what we want to do is make that biodegradable and so uh the way that gene drive actually happens uh or well one strategy i should say um is with our old friend homologous recombination and so this is the one that it's the [Music] it's the most uh it's the most difficult but it's also the most accurate and so uh so we add something to the modified gene that will target the wild type gene that we want to replace and so we'll add something that will make a double strand break in the wild type gene and so you have this double strand break here and homologous recombinations the one that only happens if you have two copies so it goes and it reads the other copy and it uses that as a template to repair this damage and since it's using our modified gene as the template for repair then it just copies it over and so the wild type becomes the modified gene and so that's that's how we do uh that's our strategy there's a few other strategies but that's our strategy for gene drive and then uh the way that we make it biodegradable is our old friend ssa and so when we build our modified gene we'll also make sure that part of what we're inserting is repeat sequences as bookends to the modification that we've made and so since there's these repeat sequences then ssa can happen and so we'll also put in something that attacks itself and so it'll cause a double strand break on itself and so when this is repaired then ssa will happen and so it chews it up makes it single strand brings the repeats together and so everything that we've added in that's modified is on is since it's in between these repeats it's in these overhangs they get clipped off and so we've gotten rid of all of the genetic the genetically modified stuff and then when it's repaired then the genetic modification is just deleted and so just sort of zooming out just looking at a population of wild-type mosquitoes okay so we we add in a small population of genetically modified males and then they spread through the population and then sort of a random the deletions will start happening and it'll send them back to wild type and so they go back to essentially wild type and so it's temporary so all right so um i guess wrapping up i just wanted to talk about some of the public the published research from the lab and so i just have just some screenshots of the titles and some of the authors and so these two papers are about the gene drive safety because when you start messing with genes and making uh genetically modified stuff and then you start talking about releasing it into the wild uh that that's a serious thing and i know i've tried to add in some jokes and make make the the uh conversation a little more light uh but that's just to hold attention and make it enjoyable uh we do take safety and uh just making sure that we're not you know irreversibly damaging the environment seriously and so there's a few papers and i'm sure you can find more about safety precautions and some of the some of the safety precautions are not even necessarily like scientific strategies it's working with local governments it's working with local charities just community leaders and just just talking to them and trying to be explain exactly what we're doing uh be open and transparent about what we're doing and uh you know different uh precautions that we're taking so uh if you wanna read more about that there's these two papers and i'm sure many more out there but these two are from our lab so and then if you want to learn more about the specific mechanism of making the gene drive biodegradable this is excellent paper and so that was that was out of our lab also and then and then i mentioned that we could do some genetic modifications but i didn't really talk about exactly what we're changing and so if you want to read more about those uh here's a three papers um this one some people in our lab discovered uh the protein that if it's expressed the mosquitoes are male and if it's not expressed then they're female and so uh and then a later paper this nix is transcriptional control over that protein and so if you make sure that nyx is expressed then the male determining factor is also expressed and so these two are basically if we push that through a population with our gene drive then all of their offspring will be male and so male mosquitoes spread the gene but they also don't bite people so they don't spread disease and they also don't lay eggs and so that's that's one of the things another thing is uh the proteins that make up the flight muscles of mosquitoes are different based on uh male and female and so we can selectively knock out the female flight muscles so the females are flightless and so a mosquito that can't fly is a mosquito that can't drink very much blood and spread very much disease but the males are still can still fly and so they can push the gene through the population and so yeah so just want to acknowledge everybody that's working on this research in our lab so our principal investigator uh boss as we call him uh is zachary edelman and then there's several post-docs in our lab i mostly work with kuhn but everybody else has helped me a lot learning the ropes teaching me about mosquitoes and learning all the all the new stuff that i didn't know before and then we have some phd students our master's student is actually working double duty because he's also one of our paid research assistants our new lab manager other research assistants our undergraduate and then also uh people that pay pay uh pay the bills and buy my dog dog food for me the national institute of health so all right so thank you so we've got a couple of minutes here that we're going to open up the floor to have some questions if you have some of those i want you to hear me say something that i hope is valuable to you everything is cumulative you heard science you heard english you heard communication you heard from his biblical perspective and an introduction you don't know that he is musically inclined everything is cumulative there is not a class that you will not take that is not meaningful and you just saw it unfold in about 45 minutes please if you have any questions let's field those at this time yes but what would be bad about completely eradicating all mosquitoes um okay so the question is what would be bad about completely eradicating all mosquitoes uh so the the joke among entomologists is the only thing that good or the only thing that mosquitoes are good for is making more mosquitoes and spreading disease but nature is complex and so uh the obvious things that mosquitoes do uh are the bad things that we know about but uh if if we just completely eradicated there may be a lot of off-target things that could happen um so i mean mosquitoes play a role in the food web they're they're food for other species some people might argue that other things can be food for for other species bats betsy more than just mosquitoes fish eat more than just mosquito larvae but we it's one of the situations where we don't know what we don't know and what we don't know may collapse an ecosystem and so we don't necessarily want to just eradicate them even though everybody just wants to eradicate them yes this doesn't really have anything to do with what you said but are they pollinators yes that yeah they are uh so since they some so the question is are mosquitoes pollinators um so since they mostly drink nectar they are pollinators so yes is so i don't know the specific science um but i i know enough to say i think so um i think it's so there there's definitely some people that mosquitoes are more attracted to than other people some people get eaten alive and some people hardly ever get mosquito bites i think it's probably more complicated than just blood type but yeah there's there's definitely some people that are much more attractive to mosquitoes than others so yes how many mosquito bites do you get and it's really part of a question tell us kind of what you do on a daily basis do you end up with actual mosquitoes around you and give us some tasks yeah so how many mosquito bites do i get like in the lab or just in general in the lab not that many but it happens so one of my first days actually being in the lab after i'd gotten finished with onboarding a mosquito landed on my arm and since it's in the lab and it's data i wasn't sure if i should kill it and so i just kind of watched it suck some of my blood and then like when i realized it was sucking my blood it was just reflex and so the curiosity left and i just swatted it and i was like actually i don't know what's the proper procedure do do we just kill him no like yeah if they're out of their enclosures just kill him no no questions just so uh it's happened a few times uh the we we don't let them just you know fly around the lab we have uh homemade stuff it's actually uh popcorn cups we'll just hot glue them together cut some holes in the side put some mesh and some different things to kind of make our own enclosures but uh yeah it's it's pretty uh homemade the i mean we have incubators that keep them at the right humidity and temperature that are what you'd expect in a science lab but the the individual enclosures that we keep the mosquitoes it it looks like a concession stand but so we we try to keep them from flying around the lab but some get out because you're dealing with hundreds thousands of mosquitoes and so it's gonna happen but we have like the electrified tennis rackets all around the lab and those are quite fun to use oh and then you asked about day to day um so uh so we rear the mosquitoes ourselves and so you saw the egg cup and so there's paper in there so uh we'll take those out and let them dry and so so this is something i sort of hinted at but didn't fully explain so the reason that we use 80's aegypti instead of some other species is they're perfectly fine if their eggs get dried out and so we'll dry out the egg papers and then seal them in like a plastic sleeve and so those are good for like three months you could probably stretch it further but we try to get a we try to hatch them out at least after three months um other species of mosquitoes like anopheles or kulex you have to just constantly just keep those going and so it's it's just it's a lot of work and it's annoying especially if it's a line that you're not actually doing experiments on and so 80s is just much easier to work with in the lab because you can store the egg papers um okay so we will dry the egg papers we'll store them we'll hatch them out so we just have like pans of water it's about an inch deep and we just drop the egg paper in there a few hours later you'll see l-ones swimming around and then usually about five-ish days later they'll start pupating and so uh we got the mosquito life cycle down like clockwork and so we usually hatch on wednesdays because then they start pupating on monday and then they'll pupate all week and we can pick the pupa and then they'll pretty much be done by friday so we don't have to come in on the weekends sometimes their life cycle is altered and so we got to come in on the weekends which is annoying but usually it's my fault anyway because i just hatched them at the wrong time or i let them get too dense because if they're if they're in the pants and there's way too many in a pan they'll grow slower so uh yeah pick pupa uh sort them by sex so the pupa are also different sizes so we have a a a machine not like a mechanical machine not like a powered machine a simple machine scott but it'll it'll sort them by size and then i have not gotten very good with that so i usually have females in with my males and males and with my females females in with the males is not so bad because you just don't use those but if a male gets in with a female pop then they're contaminated and so you just gotta throw the whole thing so i usually sort them by hand so you can tell larva you can't tell the sex but starting with the pupa and then certainly the adults you can look at the last segment on their tail and tell tell the difference between males and females and so i especially for ones for experiments i will sort those with a paintbrush and a microscope and it takes a long time but it's more accurate so so yeah just in general taking care of mosquitoes um so some of the experiments that we run we'll do injections so the way that we actually get the genetically modified stuff into the mosquitoes is uh we'll have these micro needles that we make in-house and inject embryos and so if you come tomorrow to my presentation you'll see more about how that works and then we'll do just some normal uh just lab stuff like the genotyping i mentioned we rip off their back legs we'll put that in a solution that will uh release their genomic dna into the solution then we can pipe that pipette that into a 96 well plate and do genotyping um [Music] yeah we feed them sucrose so we just have a sack of sugar and a one liter bottle and just make a 10 sucrose solution and just we have little cotton balls that we put on top of the mesh that's on the top of the enclosure and soak that in sucrose and that's how we feed them our time here has come to a close but before we uh say thank you to dr whitefield i want to remind you some of you are going to go eat lunch with us and so we're going to head that way here shortly you have the opportunity to ask many more questions there is another presentation that he will give that will be more technical so i'd encourage you to consider that i'd also consider uh hope that you will consider going to the other speakers there's a another one tonight at seven o'clock i think it would be great if you could participate in that thank you for your participation help me give dr whitefield another round