rebeccmeister: (cricket)
Projects with the big crew:

Monday night: thorough coverage of our mark-recapture areas. The weather stayed reasonably warm into the night, so teams were able to gather up a lot of crickets. I stayed behind to run a set of 9 pm crickets.

On Tuesday, we concluded that one of the paint types we'd been using for mark-recapture was not good for the crickets. Crickets painted with Testor's enamel paint were noticeably more sluggish than unpainted crickets or crickets painted with the acrylic paint we'd been using in the lab up until then. We also concluded that we were reaching a point where we weren't learning a whole lot more from repeatedly re-surveying our mark-recapture area, so we decided to switch gears for our evening plans. Oh, we also went to the beach in Santa Barbara, which was a much nicer trip than our trip to Pismo Beach last year. Not only was the beach less crowded, almost everyone actually went in the water. I should have gone for a fuller swim, but oh well.

So then, Tuesday evening, we formed four small teams and headed out in four different directions to get a better sense of the broader population structure out here. I also wanted to encourage everyone to help me collect up as many long-winged crickets as possible, because they're much more rare than the short-winged crickets and are a huge limiting factor for the circadian experiments. In order to give everyone extra incentive, we decided there needed to be a prize for the team that collected the most long-winged crickets. Casting about for ideas, I settled on a prize of a cake of that team's choosing.

It worked! Mk and I wound up hiking up a road in a valley along the southwestern part of the reserve. C and P were supposed to hike along the corresponding eastern edge, but when they reached the gate for that road, C observed two sets of predator eyes shining back at her: either coyotes or mountain lions. So they wisely stuck closer to home. B and CH headed north, and L, G, and Ms headed south.

The cake bribe worked. C and P won, and we came in second, but I still feel like a winner given that I wound up with 6 long-winged females with pink flight muscle. We set them up for a noontime circadian experiment.

As is typical for field experiments, we're making a ton of decisions on-the-fly out here, so part of the reason for trying to thoroughly blog about everything is to try and remember why those decisions were made (and also try to retain shreds of sanity because there is major Thought Tragedy of the Commons* out here).

After the noontime data collection, I became concerned that catching crickets, holding them overnight and through the next day, and then running them, was affecting their metabolic rates. The noontime crickets had higher respiration rates that are comparable to the respiration rates I've observed so far with laboratory crickets, whereas the 9 pm crickets tended to have about half to two-thirds the respiration. We aren't really aiming to study metabolism under starvation conditions, so that was a problem.

Thus, last night, we changed things up again. At 9 pm, teams set off to the two locations out of the four that had been the most fruitful on Tuesday night. Larger groups seemed prudent after the creepy nighttime predator eyes (and a note: nighttime fieldwork is a whole different ballgame than daytime fieldwork!!). I stayed back at the ranch house to prep supplies for another circadian experiment. At 10 pm, teams returned with their haul up to that point, and I wound up setting up 1 long-winged female, 6 short-winged females, and 6 long-winged males for another run starting at 10:24 pm.

At noon, I trained Mk how to help me run the experiment, so she also helped me with the evening timepoint and we turned that crank as best we could. [Interesting tidbit: a large proportion of the crickets have some sort of mite hanging out under their wing covers. I need to photograph them.] We wrapped up by around 1 am. Teams went back out at 10 pm for a second search shift, but temperatures dropped substantially last night, so the crickets weren't all that active anymore.

It's tough out here, when all our best efforts just can't quite net the numbers we need for this kind of experiment. I sort of expected that, and figure we're learning a TON out here anyway, so I'm still optimistic we'll be able to get some good papers for our efforts, even if they aren't quite what we set out to do. We shall see.

I am still feeling grumbly about that rejected manuscript, although this morning as I revisit the comments I'm coming to terms with it all. Gotta get up, dust off my fragile, bruised ego, and keep going.




*Thought Tragedy of the Commons is [personal profile] scrottie's term for what happens when one person tries to hold onto their thoughts about what they're going to be doing next, by saying that thing out loud. When they do, they disrupt the peaceful thoughts of the other people around them, who often respond in turn by voicing all their own thoughts out loud. The net effect can be complete disintegration of one's internal monologue and will for doing things, especially if one is a rather sensitive introvert.
rebeccmeister: (cricket)
After Friday's long hike (losing track of days of the week already, here...), it seemed prudent to make Saturday a low-key day. Besides, our conclusion from the hike was that we're stationed in the part of the reserve that is the most crickety, so we don't really need to go too far.

So after morning pitfall trap checks and miscellanea, it was time for another trip into town. On our first trip, L pointed out a sign for U-Pick berries right as we crossed highway 154 to take Edison Street into town. We had enough to do that time that we didn't have the capacity to stop. This time, however, we did.

And now I'm having some strongly conflicting emotions. On the one hand, I'm utterly ecstatic to get to eat fresh raspberries and apricots and blackberries, and to buy farm-direct vegetables. YAY! On the other hand, it's tremendously difficult to quell the urge to Pick All The Berries and Jam All The Things. This has bumped Sedgwick from, "Ehh, this field site is fine" to "OMG OMG I LOVE THIS PLACE." The woman running sales said that with the highway closures for the fire, traffic has been dead and she hasn't had customers, but it isn't a big deal because she has a lot of other things to do anyway.

I am trying to work on a horribly overdue manuscript review, but the exhaustion from fieldwork and dealing with logistics and lots of other human beings is making it very challenging to concentrate. In the evening, two students from a collaborating lab showed up, so there's been a lot of "Getting to know you" and such. Also a lot of intense mentoring of the current undergraduate and post-bac who used to work with me. Those are important, but draining. We're currently at 7 people. On Monday there will be a partial changing of the guard, and we'll be up to 10 people until that Thursday. The good news is that I think we're reaching a point where we'll have fairly quiet mornings and a mid-day break, which I desperately need.

We're starting to converge on a pitfall trap method. The first night, we netted a total of 1 male cricket across 5 traps. The second night, we got 5 crickets, all of which were in the 3 traps that held recorders that played back male cricket chirps and not the 2 empty traps. Last night B had the idea to take some of the males that we collected during surveys and put them out in the traps that don't have recorders in them. A team also went out and set up additional pitfall traps in another location, so we're gradually extending our reach.

Long-winged crickets with pink flight muscle are still incredibly scarce. I want to try and walk around right at dusk, but I won't get to do that tonight because I need to run the cricket oxidation procedure at 9 pm again. I'm still optimistic about working with our ongoing collection of last-instar crickets.
rebeccmeister: (cricket)
The 3 of us make for a really nice fieldwork Dream Team.

The drive down took about 5.5 hours, hitting periodic random traffic congestion, as one does in California. With an extra 1h delay at the car rental, load-up at the lab plus 3 houses, and a stop at REI for a GPS plus lunch, we didn't get on the road until 12:30 pm. Still not too bad.

I am SO GLAD I got to pack all the lab supplies. Last year was a nightmarish giant pile of stuff, whereas this year I know exactly where everything is / is supposed to be. Everything got packed very neatly into the 12-passenger van, with ample room to spare, and didn't feel like a hellish mad scramble. I have a certain hatred of stuff-piles stacked so high that things slide all over.

So far I think the cricket population density is on par with last summer. C and A got here a couple of days before we did, and in one evening were able to finish collecting what they needed, so they offered to help us. In an hour, the 5 of us collected ~80 crickets, heavily biased towards short-winged males and females. All of the 10 long-winged crickets we found had histolyzed (white) flight muscle. So we'll have to keep easter-egg hunting.

Today has involved a debriefing with one of the reserve managers, picking up some additional supplies in town, getting meals and groceries squared away for the next couple of days, setting up the full respirometry rig, and beginning to test out pitfall traps. Tonight we'll repeat our population survey (mark-recapture with last night's crickets) and will hopefully work towards gathering up crickets for some initial metabolic experiment test runs tomorrow.

-

Last year we had to stay in tent cabins because the ranchhouse was undergoing renovations and refurbishment. The cabins had healthy black widow populations (though no one was bitten), and we cooked in an outdoor kitchen adjacent to a classroom space where we worked. Showers and toilets were in a freestanding, rustic structure. It was pretty good for a fieldwork setup, all things considered.

This year, we're the first research group staying in the ranchhouse as the renovations wrap up. And OMG it is POSH. Apparently the UCSB donor who funded the project will be staying in the master bedroom on occasion. It has a panoramic view up the Reserve's central valley. It's fully air-conditioned. There were some interesting decisions during the renovation, such that certain bathroom fixtures are still adorably historic, and all of the new windows are still single-pane. Sad to see so much energy loss.

Still, there's countertop space in the kitchen that is PERFECT for the respirometry rig, and we're using the dining room for staging other projects and plotting and scheming about how to take over the world. The living room contains the most enormous television I have ever seen in my life.
rebeccmeister: (cricket)
My brain is complete mush today.

But I needed to squeeze in a Skype meeting this morning with my long-distance undergrad, who has been determinedly plugging away at learning the basics with R. We have two main tasks left with the current stage of data analysis, which we basically need to translate from English into syntax. The first is figuring out how to read in a bunch of files from a bunch of directories. This should be straightforward.

The second task is one I'd been scratching my head over. When recording cricket activity, I set up the timelapse video to record 4 crickets at a time. Our tracking software assigns individual numbers to individuals, but for various reasons individual crickets wind up having between 1 and 10 different numbers assigned to them. So, how to separate out data for each individual cricket? I'd been thinking of coming up with a method to subset the track files by the assigned ID numbers. This would require figuring out how to import a ragged data file, then figuring out a kind of complicated reference scheme.

But then today C pointed out that because the crickets are spatially separated from each other, it would probably be a whole lot simpler to just subset based on each cricket's xy coordinates. We don't expect cricket 1 to ever show up in the area occupied by cricket 2, unless something went seriously wrong with the setup (nothing did).

Foreheadslap to self. Good job, undergraduate! You rock.

This startup is a VERY good idea. I need to pay a visit.
rebeccmeister: (cricket)
Several photos from around the lab.

First, my lab succulent collection, which makes it nice to gaze toward the windows:
My succulent collection in the lab

This Granny Lamp arrived today. I was going to buy a dissecting scope for fieldwork, but got to thinking that one of these lamps would probably be just fine instead, and it might be nicer than the scope for teaching students how to dissect crickets.
A granny lamp's an entomologist's best friend

Before I headed in to work today, I paid a visit to the Albany Aquarium to see what they had in the way of air pumps. Here's the challenge: when I do nighttime cricket experiments, I need to keep the crickets in the dark / under red light, up until the very end of the procedure. We have a small room that doesn't have too much light pollution, but it's too small for the whole benchtop respirometry rig. See, it's full even when I just have injection/blood collection supplies laid out:

Daytime circadian setup

(nighttime view):
Nighttime circadian setup

Here's the scale of the benchtop respirometry setup, for comparison. Too much stuff to fit in that small, multi-use room.
Li-Cor / Oxilla stop-flow respirometry setup

Previously, I was able to use a second respirometry rig that's built into a Pelican case and designed for field measurements, but a grad student in the lab is going to take that rig out into the field this summer, so it's off-limits now.

The people working at the Albany Aquarium Store were knowledgeable and helpful, and helped me find an adjustable aquarium pump that provides enough air flow, but not too much, for the purpose of flushing dry, CO2-free air through the syringes that house the crickets during the circadian procedures (at ~800 mL/min, in case you wondered - lowest setting on the pump). Perfect!
Aquarium air pump CO2 flush setup

Okay. To round things out, here's a sophisticated gadget I designed myself. It's a cardboard tube, modified to help keep the cricket syringe upright in a hands-free manner, so I can do other things while the syringe is being flushed:
Sophisticated syringe holder

Time to design something similar for the aquarium pump setup.
rebeccmeister: (cricket)
One of my undergrads has just completed her honors thesis on flight in the California wing-dimorphic cricket species, and after a tremendous amount of work she wound up getting some really cool results. Based on a prior publication, we all thought that making female crickets fly for 10 minutes would cause them to histolyze their flight muscles and grow their ovaries.

But that wasn't the case. More of the flown crickets actually retained their flight muscle compared to tethered, unflown controls (the tethering controls for the handling required to get the crickets to fly). There was a small trend towards larger ovaries in the flown vs. tethered crickets, but it was not significant and was much smaller in magnitude than the difference between pink-muscled crickets and white-muscled crickets.

Of course, these findings raise a billion new questions, but this was a fantastic and interesting project by itself.

Somewhat related to all this, something cool happened on Monday night, when I stayed up late to complete a late timepoint and then slept overnight in the lab. At the moment, I'm running assays to check the hemolymph composition of the California cricket species at different times of the day and night. We want to study processes in crickets that are in a post-absorptive state, so I take food away from the crickets 3-4 hours before I start the experiment. Logistically, that means that I transfer crickets from a co-housed sweater box into pint-size deli cups that contain a cotton ball (so they aren't dehydrated). [I'll try to remember to take a photo of the setup this evening.]

Anyway, to estimate a cricket's total hemolymph volume, I inject them with fluorescently-labeled inulin, let them sit for 30-60 minutes, then puncture the exoskeleton and collect the hemolymph. By measuring the relative dilution of the inulin, I can back-calculate to the total hemolymph volume.

One of the things I discovered when I was developing this method for crickets was that the inulin is very difficult to dissolve in water, and it comes out of solution easily. So to ensure that my results are as consistent as possible, I vortex the tube of inulin every time that I go to draw some out for an injection.

I'm using an ancient, inherited vortexer that rattles around a whole bunch, and thus vibrates the containers holding the crickets.

I'm not sure if it was the vibrations from the vortexer, or simply the time of night, but on Monday night I saw at least three of the ten long-winged crickets open their wing covers and start vibrating their hind wings, which is a preparatory behavior for flying.

In comparison, I've never seen the Florida crickets do this.
rebeccmeister: (Acromyrmex)
I'm currently reading the chapter on insect feeding and digestion.

Here's the thing: insects are tremendously diverse. They are one of the most diverse groups of organisms on our planet, and that's a big part of why a lot of people think we need to do a good job of cataloguing them (to say nothing of preventing species from going extinct).

It naturally follows that insects have diverse, amazing feeding apparatuses and digestive systems. I'll just highlight two.

First example: mosquitoes. People like to refer to mozzies as "flying syringes," because they don't simply poke you and then lap up blood as it comes out. No: they inject anticoagulants, and THEN they take out blood.

Then there are xylem-feeding insects, which extract liquid from a fluid-filled plant tube that operates with negative pressure (in contrast, blood flows via positive pressure). To suck out the liquid, these insects have to have a strong, muscular pump inside a region of their head called the cibarium. Scientists think the need to fight against negative pressure explains why there aren't any small xylem-feeding insects: those insects wouldn't have adequately large cibaria to counteract the xylem's negative pressure.

Yeah, crickets are pretty boring in comparison.

There are also lots of exotic modifications to the digestive tract, depending on whether the organism is trying to extract nutrients from an extremely nutrient-poor aqueous solution, like the xylem-feeder, or from a slurry, et cetera. The insects that feed on extremely nutrient-poor liquid solutions tend to have incredibly long and convoluted digestive systems that promote swift removal of water and excess nutrients. Mosquitoes, for example, pee out extra liquid while they feed (starting at 2:02). Again, way more exotic than standard mammal examples.

The thing I really hadn't thought about much is this: digestive enzymes are very effective, but don't necessarily discriminate between the lining of the gut and the food that's getting churned around inside the gut. Therefore, to prevent self-digestion, most organisms very carefully regulate the production of digestive enzymes, tuning production to both the timing and composition of the food they've just eaten.
rebeccmeister: (cricket)
I feel like this piece does a nice job of capturing some fundamental points about the roles of science in society. It not only does that, it also disentangles some points about benefits and drawbacks to large-scale institutions, public and private.

This article makes some related points, although without the rich historical context.

I plan to join the march tomorrow, carrying a sign that says, "Invest in public good!"
rebeccmeister: (cricket)
Two interesting things that popped up on the Tweet-machine today:

A paper was recently published in the journal Nature showing how groundwater depletion is linked to international food trade. The first figure shows geographic patterns of groundwater stress and which crops are grown. Let's just say the American Southwest isn't looking great. Figure 3 shows how much goes where, and Extended Figure 1 shows historic data from 2000. Fascinating.

Secondly, apparently octopuses and other smart cephalopods employ way more RNA editing than researchers anticipated, compared to all other organisms studied to date. To me, this is a great example of why it's important to study a diverse array of organisms. Nature is more creative and clever than you might think.

Agog

Mar. 24th, 2017 01:12 pm
rebeccmeister: (cricket)
Today I learned that the LUNGS make blood.

What.

Just pause and think about that for a moment.

Where did you think blood was made? The bones? Ahaha. Outdated knowledge now.

I'm reminded of learning about a newly discovered organ in the human body.

These are the kinds of things that need to be taught in college Introductory Biology courses. I feel like this could wake students up to the importance of ongoing scientific inquiry.
rebeccmeister: (cricket)
Yesterday: got to lab by 9 am so I could spend regular working hours working with some people from the Lawrence Berkeley Labs who are making a science film about crickets for middle school students. I was a little bummed to learn that the film won't be publicly available because they will sell it as a science curriculum tool. But on the other hand, people creating educational material need to be compensated somehow, so I suppose this is one method of ensuring that happens. And besides, the person in charge of the project said she'd share the macro video footage of crickets eating and laying eggs. They rented a fabulous macro lens for the project. Also, the project was a fun change of pace.

After that, K and I did my normal Thursday activity of caring for our cricket stocks. You can't do experiments if you don't have crickets, after all.

Then we stayed up until midnight watching episodes of The Big Bang Theory, which I'd never seen before. Can I just say it is so weird to see the academic lifestyle turned into a Friends-style sitcom? But also, I laughed.

At midnight, we successfully finished out the cricket hemolymph project. So that felt satisfying. I made it to bed by 2:30 am.

I dragged myself out of bed this morning at 8:15 so I could go to a talk by my friend ZS. He works on the same cricket system at a nearby university, and has also started doing some experiments with ants, so we have a lot in common. Even better, his research questions and approaches are complementary to mine.

Early this afternoon, we got results back from our circadian tracer experiments. K has calculated that since the beginning of October we'd run 99 crickets in a total of 12 circadian sessions, which each last 3-3.5 hours. So that's 36 hours of intense work, not counting the time it takes to sort and set up crickets every day, analyze data, prepare and troubleshoot, and come in at ungodly times to take away food or run experiments.

That said - I made some very preliminary plots of our results, and they look good and interesting. So after weeks of working really hard and not really knowing whether that work was accomplishing anything, we have results.

I really want to rest, but I should really work on a job application instead.
rebeccmeister: (cricket)
Yesterday morning left me feeling sufficiently terrible that I left work at around 1:30, went to Berkeley Bowl to pick up some groceries and cricket food, then went home and curled up on the couch with the cat. Cat therapy.

This is one of those head colds that starts with a scratchy back of the throat. I am pretty sure that the only cure is rum.

[livejournal.com profile] scrottie and L are both sufficiently civilized human beings that they came home with candy to give out for Halloween. I was feeling sufficiently antisocial that I didn't want to repeatedly answer the door, so we set out one of those plastic pumpkin buckets and periodically refilled it. Seemed like a reasonable compromise. I feel bad about missing out on an opportunity to get to know more of the neighbors and see kids in cute costumes, but at least we contributed to the collective sugar-high. And I didn't pass on my cold.

This evening we'll do another 9 pm timepoint, and then we'll ship out a box of samples in the morning. Then we'll take a couple of days off while we wait to find out the results and whether we need to continue re-tooling things for the glutamate and oleic acid tracers. But just for fun, we'll probably stay up until 1 am on another night this week anyway to finish collecting data for a different project (characterizing cricket hemolymph across the circadian cycle). Given that we finally have good cricket numbers, we might as well put the crickets to use.
rebeccmeister: (cricket)
Here is a grid of sample sizes for the circadian experiment:

Sample sizes.jpg

We haven't been able to decide which is worse, the 1 am timepoint or the 9 am timepoint. I'm leaning towards the 9 am timepoint. You'd think 9 am doesn't sound all that bad, right? But we have to make sure the crickets are in a postingestive state, so someone has to take away their food 4 hours before the timepoint. And the 9 am timepoint actually starts at 8 am, so that means getting to campus by 4 am.

Even morning people have limits, and my limit is having to wake up before 4 am.

Also, when the timepoint starts at 8 am, that means we have to prepare certain things right before the experiment itself starts, so the work actually begins closer to 7 am.

To top things off, the person who has been helping on this project, through thick and thin, lost track of things and slept in this morning. As a result, I learned that I *can* run the whole experiment by myself. That said, I really, really, really hope I never, ever have to do that again.

Those glucose samples are crossed out because of a snafu with our respirometry setup. I have to manually set the range for our high-precision carbon dioxide detector, but early on I set the range too low and so our measurements of the amount of carbon dioxide exhaled are underestimates. We might be able to get that info from our collaborator who measures the C-13 sample enrichment, but I'm not certain of that yet.

Also, my Halloween costume is made of polyester, and polyester smells awful, especially when one is sweating from all the extra anxiety and running around.
rebeccmeister: (Acromyrmex)
Full day of talks today.

I set my smart-o-phone for a 6:30 am alarm, to have plenty of time to get up, make coffee, eat breakfast, then head over to the hotel for the pre-conference conference of the North American Section of the International Union for the Study of Social Insects (NAS-IUSSI).

I woke up at 7:20, in a panic because I was so late.

I made it to the first part of the meeting on time, but in my panic I forgot to bring my poster with me. So I had to miss an hour of talks to go back to the hotel and get it.

Other than that, it was wonderful to see so many friendly faces, old and new, and to hear about all the different kinds of questions people are asking and addressing in social insects.

It also makes me glad, though, to be doing a mixture of work in solitary and social insects. Working just with social insects is a bit too insular, methinks.

Status

Jul. 25th, 2016 05:16 pm
rebeccmeister: (bikegirl)
Sleep deprivation is catching up with me, hard.

Friday night/Saturday morning: Up until 3 am to complete our last glucose circadian trial (samples will be shipped off this week for gas analysis). Then up again at 9 am to finish packing and get into the lab, pack more, and head out to Sedgwick.

We reached the field station just before 6 pm on Saturday. Traffic wasn't horrible but we hit a couple of slowdowns somewhere south of San Jose (Gilroy), so we followed the Goog's suggestion to hop off Highway 101 and onto Highway 25, past Pinnacles National Park, which made for some nice sight-seeing en route.

There was an organic grocery store on the highway about a five minutes' drive from the reserve, so we picked up supplies for a simple supper. By the time that wrapped up, it was starting to get dark. As we walked from the dining area towards our tents, I encountered the first female crickets. It was time to start cricket-hunting.

I believe we were up until somewhere around midnight that first night. Apparently, the crickets were much more abundant this year than last, seeing as we were able to collect close to 100 crickets in only two hours between the six of us (five adults, one five-year-old). So we were able to quickly accomplish our first aim of collecting enough crickets to replenish the lab stocks. With that task out of the way, we were then able to focus on a second goal of assaying cricket hemolymph and body composition for crickets in the field.

I managed to sleep in until around 7:45 am on Sunday. We spent the morning regrouping and generating plans for Sunday night, then drove out to Pismo Beach to dip our toes in the ocean in the afternoon.

For night #2, we conducted two activity surveys, one between 9-10 pm, another between 4-5 am. So I slept from around 11 until 4, then from 5-6. At least we learned that we'll never have to get up at 4 am again because the crickets were no longer active at that hour.

At 6, we got up, packed and cleaned up, and hit the road by 7:30 so we could get back to campus by 12:30 and one of the undergrads could make it to her course.

Now I am unpacking and repacking to head to the Midwest tomorrow morning for vacation, so to speak.
rebeccmeister: (cricket)
The science jet lag isn't so horribly wretched at the moment, although I'm expecting some carryover through tomorrow.

We did the first run of the 1 am timepoint last night. That meant hanging out in the lab until 8 pm, when it was time to remove food from the cage, then hanging out until 11:30 when it was time for our final preparations, then running the show from 12-2 am.

To fool the crickets into thinking it's nighttime, we worked under red light. K took some photos at my request:

Nighttime tracer injection setup

Nighttime tracer injection setup

Nighttime tracer injection setup

Nighttime tracer injection setup

We made several mistakes, which could mean having to do a three-peat of this timepoint. At some point, the light to the left of my head fell down and the bulb burned out. While plugging in a replacement light, K must have accidentally jostled the adapter for the fan pump we were using to flush those 60-mL syringes with dry, CO2-free air. Unfortunately she didn't notice quite when that happened, so we had to re-flush all of the syringes and start the stop-flow incubation 30 minutes late. We'll still be able to compare long-winged versus short-winged crickets, but since we don't know the full timecourse for our injected tracer we don't know if these results will be comparable to the rest of our results.

At one point, I also opened the door and exposed the crickets to some incandescent light. Argh.

Then I slept on C's office floor in a sleeping bag until 4 am, when I got up to remove food from another set of crickets for the 9 am timepoint. This system wound up being less hellish than just running one of the two timepoints. Once the crickets were squared away at 4 am, I slept until 7:10 and then walked over to Yali's for a latte and scone. We wrapped up the 9 am timepoint by 10, and I headed home for a couple of hours.

The downside of this kind of arrangement is trying to figure out what to do from 10 am - 2 pm. I'm not very good at taking naps, nor do I want to completely throw my sleep schedule out the window. So I rested and worked on a couple of household chores.

Tomorrow we only have a couple of crickets that will be ready, so we'll take things easy and just run the 5 pm timepoint. We'll be doing another 1 am stint on Friday night. We're actually making very good progress overall, although things can get a little frustrating because we lose data left and right due to various small mishaps. It's just a difficult experiment to complete smoothly.
rebeccmeister: (cricket)
Over the last couple of days, I keep thinking about justifications for various kinds of experiments, and the impact of those experiments. It's necessary to consider justifications and impact, especially when one studies organisms that might be dismissed or ignored.

The circadian metabolism project, to me, is inherently cool and interesting because of the uniqueness of the system we work with. The vast majority of research on hormonal differences focuses on huge differences between males and females, but those differences are very extensive and far-reaching. The other major alternative is to work with a well-developed model system where individual genes can be manipulated, but most such systems lack clear connections to natural settings. The closest related work occurs in honey bees, which are still domesticated insects. We don't want this line of work to be pigeonholed as something that applies only to crickets. So we continually have to ask ourselves about the broader implications.

The circadian metabolism project is moving forward nicely. From my initial round of hemolymph collections, we've learned that there are constitutive differences in the total hemolymph volume between the long-winged and short-winged crickets; the long-winged crickets have a lower hemolymph volume across the day. Hemolymph volume also changes in both morphs over the course of the day, increasing towards nightfall. Right now an intrepid undergraduate is working to measure lipids, carbohydrates, and protein in the same samples, so we'll have a good and detailed picture of how these nutrient levels fluctuate across the day, too.

The feeding experiments...progress. I'm just starting the next round this week. The first round was informative but left some large questions unanswered. At first, I was concerned that it looked like the adult crickets might just be feeding randomly when given a choice between two diets because their intake was pretty evenly divided between the dishes. However, the last-instar crickets definitely made a different selection, preferring carbohydrate-biased food over protein-biased food. Perhaps it is unsurprising that juvenile crickets have different nutritional requirements than adult crickets, but if you were to have askd me before this experiment, I would have predicted that the juveniles would want more protein than the adults, not the other way around.

There have been two major confounding factors, however: out of the 34 juvenile crickets I set up, 14 died, and only 2 crickets (both male) developed into long-winged adults. In contrast, the stock colonies tend towards 50-50 production of long-winged and short-winged adults, and only one of the 54 adult crickets died. Supposedly morph fate is locked in by the time crickets reach the last instar, so this could simply reflect the luck of the draw, especially if the short-winged crickets are slower to develop than their long-winged counterparts. Or it could be that something about the diet was inadequate for the long-winged juveniles; the stock diet is more nutrient-rich in protein and carbohydrate, and contains more fat as well.

I've also been interested in determining how group size affects feeding, so for the next round, we're also setting up groups of four crickets.

There are a lot of large unknowns when it comes to how nutritional needs vary across life stages in different kinds of organisms, and about how flexibly organisms can respond in terms of nutrient allocation when confined to suboptimal foods. And we don't know a tremendous amount about how crickets manage nutrient intake in the wild. All of this would be useful to study from a comparative standpoint, because the vast majority of related work has been confined to vertebrates. To what degree are vertebrates weird and special, vs. sharing things in common with smaller-lived, more ubiquitous organisms? There have been heated debates about this recently in the ageing and lifespan literature.
rebeccmeister: (Acromyrmex)
While faffing around today in between productive bouts of working on the leafcutter manuscript (!), I came across a link to this article by Kieran Healey on social media and sociology. The excerpt posted on the Dynamic Ecology blog has convinced me to read and consider it further, and actually, it's relevant to the leafcutter manuscript in addition to being relevant to the act of blogging. Here's are some of those tantalizing excerpts:

"Here is also a natural connection here to the world of scholarly research. Although by now thoroughly professionalized, academic life has deep roots in the desire to talk about scholarly preoccupations in public, and in one’s spare time. It is in this sense an aspect of civil society. On a personal level, having the desire to go and tell people about your work is a good a sign that you are substantively absorbed by what you are doing. The point generalizes to disciplines. To the degree that thinking, talking, and arguing about research in one’s spare time and in public is a feature your field, it is a sign that your discipline is confident about what it does. Modern social media brings together these shared features of civil society and academic discourse in a new way. Social media platforms facilitate and accelerate the possibilities for talking about one’s
work in public, assuming we want to take advantage of it."

"In “Science as a Vocation”, Weber remarks that although we do not get our best
ideas while sitting at our desks all day doing regular work, we wouldn’t get any good ideas unless we sat at our desks all day doing regular work. In a similar way, successfully engaging with the public means doing it somewhat unsuccessfully very regularly. This fact is closely connected to the value of doing your everyday work somewhat publicly. You cannot drop a lump of text onto the Internet and expect anyone to pay attention if you have not been engaging with them in some ongoing way. You cannot put your work up on your website, or “do a blog”, or manufacture interest in your research like that. There is a demand side as well as a supply side to “content”, and most of the time the demand side does not care about what you have to say. This is why, in my view, one’s public work ought to be be continuous with the intellectual work you are intrinsically motivated to do. It is a mistake to think that there is a research phase and a publicity phase. Your employer might see it that way, but from a first-personal point of view it
is much better—both intrinsically and in terms of any public engagement you might
want—to think of yourself as routinely doing your work “slightly in public”. You write about it as you go, you are in regular conversation with other like-minded researchers or interested parties, and some of those people may have or be connected to larger audiences with a periodic interest in what you are up to."


...and so on.
rebeccmeister: (cricket)
In addition to finishing my first quilted ant, today I set up my first enzyme kinetics assay. We have a whole list of different enzymes we want to examine, but we have to start somewhere, so I figured we'd start with what should be the easiest, citrate synthase.

In preparing for this assay, I came across a paper from 2001 where a group of exercise physiologists set out to look at the effects of acute exercise on citrate synthase activity in trained vs. untrained human skeletal muscle. Here's how the study worked, in a nutshell: they took a set of six guys (yeah, these things are always on guys, right?) who were initially characterized as "sedentary." In the initial phase, they had the participants perform an "acute knee-extensor exercise bout with their left leg," and then right after that they took muscle samples from both legs and measured citrate synthase activity. Then they had these participants complete an individualized 8-week left leg training regimen with their knee extensor machine (1-hour sessions, three times a week). Then they repeated the acute exercise bout and took another set of muscle samples.

I am going to refer to this paper as "the skateboarder study" because I once read about a guy who skateboarded across Australia and wound up with one leg/calf much stronger than the other. I hope those guys were compensated well for the whole procedure.

Anyway. My mentor from Nebraska gave us a ton of advice for how to validate our enzyme assays, but it's one thing to read through the methods for these things and a completely different thing to actually sit down and go through the procedure. Basically, as soon as you go from living tissue in an animal to a sample in a test tube, you have to be very careful about how you're handling things to ensure the sample remains representative of what happens in the living animal. In my case, that meant assembling a game plan for: dissecting crickets, quickly homogenizing tissues to extract enzymes, mixing up the reaction cocktail, and measuring the reaction under precise conditions. Oh, also, all of the reagents involved are expensive and come in tiny quantities, so you have to know how to work with small masses of things without royally screwing up. No pressure, eh?

Long story short, my first attempt was about as successful as it gets. I ran both a negative control that stayed negative (no reaction occurred in the absence of the enzyme), and a positive control that was positive (reasonably good reaction when I added a known quantity of expensive purified enzyme). In addition, I ran some samples from two crickets. The cricket flight muscles behaved beautifully. The cricket fat body samples did not behave, nor did the leg muscle samples. Puzzling, but not insurmountable.
rebeccmeister: (cricket)
Things to be working on:

Experiments:
1. The circadian metabolism project. One of TZ's key discoveries has been finding a difference in circadian hormone patterns between the long-winged and short-winged crickets. Prior to this finding, most people had just focused on studying constitutive levels of hormones. However, thanks to careful and extensive measurements, TZ found that there are important differences in the levels of a key insect hormone known as juvenile hormone (JH) between long-winged versus short-winged crickets, at different times of day. In the long-winged crickets, JH titers in the blood fluctuate and reach a peak right at sunset. The short-winged crickets experience minor fluctuations. When hormone levels differ like this, one logical question is, what is that hormone doing? One logical hypothesis is that hormone levels are tied to flight, which primarily happens at night, and only in the long-winged crickets. Flying is likely to involve shifts in metabolism, so we're working to determine how this happens.

There have been hitches. One big logistical hitch at the moment was something of an unknown demon until a day or two ago. Administrative changes and resulting high turnover are causing serious slowdowns in ordering and receiving supplies. I'm having to track down the right people to prod. I have three separate categories of things on order for this experiment, and am tapping my foot as I anxiously wait for them to FINALLY show up.

2. Cricket feeding and respiratory metabolism project. We're trying to get things up and running with a new cricket species here in California, Gryllus lineaticeps, because of concerns about the consequences of working with cricket stocks that have been kept in the lab for 20-plus years. They are developing slowly, slowing things down. The respirometry part is the most rewarding at the moment, because the respirometry setup is here already and running pretty well. Plus there are interesting differences between this species and the one I've primarily worked with up until now (Gryllus firmus).

Writing:
1. Leafcutter manuscript: I've asked my PhD advisor to take a crack at revising the Introduction to this paper because I've been bashing my head up against it without making much progress. In the meantime, new publications keep appearing that are relevant (need to keep up with the literature!), and I need to work on the Discussion. I keep waking up at night thinking about this one.

2. Cricket amino acid metabolism: This is the one I'm crunching away at the most, at the moment, because I'm in a productive phase of the data analysis process that doesn't yet involve too much thinking.

3. Cricket lifespan/reproduction manuscript: I've put this one on hold for the moment because it's too much to think about and I would lose too much time if I tried to work on it simultaneously, because task-switching.

-

I also need to review a manuscript on a subject very closely related to the leafcutter manuscript, that is making me nervous about the leafcutter manuscript.

So, I have plenty of things going on, but it's somewhat tricky to figure out how to focus and organize my time. Not a terrible problem, really.

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