Small fish, big questions - using self-guided guppy kits to teach evolution

Observing how predators preferentially chose
guppies from certain environments
(guess which ones?)

I have been involved with a major outreach project to create self-guided kits for teaching evolution in K-12 classrooms. This project is a collaboration between me (as part of my broadening participation efforts on my NSF grant), Dale Broder (a senior graduate student whose work we are building off of), Dr. Cameron Ghalambor (who has helped Dale implement a similar program with local 7th grade classrooms), Dr. Lisa Angeloni (who has helped with several other similar outreach projects), and the CSU College of Natural Sciences Education and Outreach Center (yes, we have one of those!). The goal for these kits is three-fold: 1) to be able to expand our program beyond what we can reach ourselves (this is where the EOC comes in!), 2) to increase classroom engagement by using live animals, and 3) to use an authentic science approach to increase scientific self-efficacy (help students realize they CAN actually do science) and learn about evolution the same way scientists have done. We are particularly interested in implementing this program in schools that serve low-income and underrepresented students. To address potential psychological barriers these students may have (which they may not even realize), we have made a significant effort to include scientists representing several aspects of diversity, both in an introductory video the students will watch prior to starting the activity, as well as with an interview included in the final pages of the instruction booklet. These kits will be directed toward middle and high school students initially, but could also be adapted for other age or ability levels as well.

Follow along here as I post updates on how these kits are coming along!

UPDATE 9/17/15
Well, as far as an update goes, this is a little late as we have made significant progress on the kits since we first met with the EOC back in May. We have also submitted an abstract for a poster at the upcoming Society for Integrative and Comparative Biology meeting in Portland, OR (see my previous post).

The activities for the kits include:
1) Observe the variation in physical differences between males from three populations of guppies (pet store, wild low predation, wild high predation).
2) Understand how color patterns are passed on between parents and offspring using punnett squares.
3) Observe guppy predator avoidance behaviors and quantify survival when exposed to predators (this will be done using pre-recorded high-speed videos).
4) A thought exercise to address how color patterns change over time due to various types of selection.

In the last few weeks, Dale and I have figured out how to assemble small acrylic tanks that don't leak (not a minor task). These will be used as small observation tanks.

Dale gluing a tank

The finished product!

I have also been filming Catalina, one of our pike cichlid predators, capturing guppies. She has a choice between representative males from each of the populations. I used a go-pro to film at 120 fps but the video below is from my Nikon DSLR (not high speed).

We have also been working hard on assembling the introductory video, with the help of a local videographer. This included recording an interview with each of our scientists using some local news crew equipment that Dale borrowed.

Interviewing Cameron in the breeding lab

If that wasn't enough, we recruited an undergrad, Ty Fiero, to help with 3D printing some guppy stencils, so the students can draw a fish that they can then add fins to and color in.

Yes, those are glow-in-the-dark!
I drew them myself!

We have also been working closely with Andrew Warnock, from the EOC, to compile the instruction booklet that the students will use. This included coming up with a name for our kits.

It's coming together! Our goal is to be finished in time for a teacher workshop in October.

UPDATE 09/18/15

We now have predator silhouettes for students to test how each guppy might respond to predators. They will then make predictions about what they think will happen with real predators. I made a range of sizes to figure out which one works best.

UPDATE 10/29/15

So we didn't make our October deadline...

We ran into some hurdles with IACUC approval as well as trying to figure out better options for the observation tanks. We've decided to change the teacher workshop to January, which will actually work out better for when they will teach evolution in their classes. Plus it gives us a little more time to make everything just right.

Today we were finally approved to move fish to the Education and Outreach Center!  We had to set up the tanks first without fish so they could inspect the room. Now that we're approved, I added some living things today - some nerite snails and some aquatic plants, Vallisneria and Ludwigia. Next comes guppies...

20g tank for petstore guppies

10g tanks for wild guppies

UPDATE 12/10/15

We're making the final push for our guppy kits! Dale and I have put a lot of work into the instruction booklet, and just wrote the final pages outlining the 4 components of Evolution the students worked through (Variation, Inheritance, Selection, and Time). We also provided some FAQs in case students have more questions about the projects they do.

We also finalized our introductory video that the students will watch before they start. This will be used to introduce the guppy system and let them hear from real scientists.

We encountered a hurdle recently regarding the observation tanks in the kits - do we ask the students to observe one guppy at a time and rotate, or do we ask the students to observe multiple guppies at a time, and if so, how do we facilitate that? Since variation is an integral concept for students to understand, we decided we wanted the later option, so then the question became do we put multiple fish in one tank or do we include multiple tanks. One of our collaborators had the idea to make nesting tanks, so we gave that a shot! They turned out well, so I think this is what we will go with.

Two nested observation tanks

 We also encountered a problem with the most recent set of 3D printed stencils - that a pencil wouldn't fit in the lines. So we asked Ty to print it one more time, hopefully for good this time. As of 10am this is what they looked like:

3D printing guppy stencils (Photo by Ty Fiero)

Here are the completed stencils, and what the traces look like:

The guppy stencil finished product!

This is going to be AWESOME!

The last piece of this puzzle that I have been working on is the poster that I will present at SICB this January. Dale and I will be co-presenting our poster (abstract #P1-7) on

Monday Jan. 4 from 3:30pm-5:30pm in Exhibit Hall A

Also, if you're interested in hearing about the work that led up to the projects we included in the kits, Dale is presenting a talk (abstract #110-3) on

Thursday Jan. 7 at 9am in room B115 

UPDATE 12/18/15
The first full draft of the instruction booklets have been printed!

Andrew did the illustrations!

We also now have an "official" kit pelican case! Blue is the perfect color for something with fish!

It looks like a kit now!

Everything fits inside!

AND...the poster is finished! SICB 2016: ready or not, here we come!

Well I guess you'll just have to come see our poster!

Updates have been moved to a permanent page, which can be accessed here: http://emilyakane.blogspot.com/p/guppy-kits.html

SICB 2016 Portland, OR

I will be contributing to two abstracts at the Society for Integrative and Comparative Biology meeting this January in Portland, OR. Both are less traditional for me, so I look forward to hearing feedback/comments. Here are the abstracts (dates/times will be updated when they are available):

Dale presenting about guppy outreach
at "Science on Tap" Fort Collins

First, Dale Broder and I will be presenting a poster in an undergraduate biology education session describing our most recent endeavor with guppy outreach:

Using self-guided “guppy kits” to teach adaptation and evolution with authentic science

Emily A. Kane¹, E. Dale Broder¹, Andrew C. Warnock², Courtney M. Butler², A. Lynne Judish², Lisa M. Angeloni¹, Cameron K. Ghalambor¹

¹Department of Biology, Colorado State University

²College of Natural Sciences Education and Outreach Center, Colorado State University

The concept of evolution is central to our understanding of organismal biology, but the United States has a poor understanding and acceptance of evolution compared to other countries, which is potentially influenced by the limited availability of evolutionary biologists that can assist teachers when covering this subject. We want to fill this gap by making our knowledge and resources easily accessible to teachers. Additionally, we want to use authentic science and hands-on experimentation to reach students who are not comfortable with the concept of evolution. We have designed a self-guided activity that utilizes live Trinidadian guppies (Poecilia reticulata) to explore adaptation, selection, and evolution. These “kits” include a video introduction by the researchers, activities observing differences in color and survival between 3 populations (domesticated and wild caught low- or high-predation), and a booklet providing guidance on the formation of hypotheses and conclusions. These kits can be used by the researchers for outreach events sponsored by the university, but can also be borrowed by local K-12 teachers to be performed independently, thereby enhancing the ability to reach a greater number of students while maintaining the benefits of a small-scale program. Similar activities performed previously demonstrate an increased retention compared to traditional lessons. Therefore, our goal is to use these kits to supplement local K-12 education, particularly at schools with under-represented populations. 

***UPDATE: Our poster will be on display Monday 1/4 from 3:30-5:30pm***

Delaney posing with the first bullfrog
she ever caught in the wild

Second, I have been working with an undergrad honors student, Delaney Laughlin, who will be presenting a poster of her honors thesis work:

Complexity, flexibility, and success: The role of feeding behavior on competition between native leopard frogs and invasive bullfrogs

Delaney N. Laughlin¹ and Emily A. Kane¹

¹Department of Biology, Colorado State University

Invasive species are a global epidemic that has a significant impact on the survival of native species. Bullfrogs are native in much of the United States, however, they have been introduced in a variety of habitats both west of the Rocky Mountains as well as on other continents. We seek to understand one potential mechanism by which they are able to succeed in these variable habitats, specifically how foraging success is influenced by the coordination and flexibility of feeding.  We predict that there will be a greater difference in success rate between bullfrogs and leopard frogs when capturing prey from variable substrates. These differences between species might be because bullfrogs exhibit a greater complexity and coordination in movement, and this specialized prey capture behavior may increase success across multiple habitats. Alternatively, reduced complexity and coordination of movement could lead to increased success in bullfrogs because this generalized prey capture behavior permits flexibility. To test this idea, we will use high-speed video to record foraging behaviors of leopard frogs and bullfrogs from a terrestrial and aquatic substrate as they capture a suspended live cricket. Coordination will be calculated as the correlation among timing variables describing movement of body parts (i.e. hindlimbs, forelimbs, mouth, tongue, etc.). Success rates between the two environments will indicate the flexibility of each species.  This information could be vital for providing ecologists and conservationists a new perspective of the mechanism contributing to how invasive species are able to persist in novel environments.

***UPDATE: This second poster has been cancelled due to unforseen circumstances***

Society for Experimental Biology meeting, Prague, Czech Republic

That's Prague Castle in the distant left

I have just returned from Prague, where I presented my research on modulation of integration in bluegill sunfish. I have been told that SEB is "the European SICB", and I wanted to go to this meeting because there are lots of researchers in Europe that never make it to the meetings in the US, so I wanted to learn more about what's going on over there.

The first thing I noticed about Europe was from the air - there are lots of farms, but none of them are squares/circles like in the US.  It was interesting arriving in Prague, where the Czech language was so different from most of the ones I am familiar with.  However, some words were still recognizable, like "toalety", which, based on it's location, I assumed meant "toilet".  There were also other things that seemed strange to me, like how the bed covers didn't span the entire bed, and how the shower only had a partial glass door so water got all over the floor. However, some things were still quite familiar, like the american pop music playing in the driver's car on the way to the hotel from the airport. In terms of food, the one thing I noticed was that Czech food = meat, potatoes, and cabbage.  There is little in the way of vegetables, especially the leafy kind. I found myself craving a salad when I got home.

Flight over Czech Republic, nothing is square

Flight over Colorado a week earlier,
everything is square/circle

Yes, that comforter is the only bed covering,
and it's too small

Shower with only a small,
partial glass door

The conference lasted 4 days, and included events like the "Women in Science Dinner" and several keynote lectures.  There was a large Biomechanics presence, and each day had a fair number of talks. I was really proud of my former labmate and current friend Kathleen, who gave a stellar talk on modulation of muscle mechanics in anoles! I also gave my talk, which received some interest on Twitter, mostly because people liked my cool high-speed videos. There were also some interesting talks in other sessions about things I didn't know I would like, including icefish genetics as it relates to anti-freeze proteins.

Kathleen Foster, presenting at SEB

A tweet about my talk

Another great thing about the SEB meeting is that I met so many people from so many different countries, including Australia, France, England, Scotland, Portugal, Lebanon, Canada (including Quebec, which is still Canada), South Africa, Germany, and Poland.  I was joking that I was going to leave Prague with an accent, but I wasn't sure which one. It was really interesting hearing everyone else's perspective on challenges that women (and others!) face in academia, as well as other aspects of academia.

Great group of scientists chatting over dinner

The conference kept me extremely busy, and it wasn't until the third night that I actually made it out of the hotel.  Since breakfast was served at the hotel, and the conference served lunch and several dinners, there was no reason to leave.  It was nice to finally get out and explore the city a bit with Ola, Kathleen, and some students from Cambridge and the RVC. We grabbed some dinner at a place that served local cuisine and then wandered down to the Charles Bridge (after acquiring some ice cream, of course!).

The Prague metro

Imagine that, meat and potatoes

Having some ciders,
that's right, out of a straw

The Atomic Clock

On Friday, Ola, Kathleen, and I decided to skip the morning talks to have some catching up time for the three of us, since we hadn't seen each other in almost a year. We headed back downtown and spent several hours walking around the main part of the city.  I couldn't believe what amazing architecture there was!  It felt like I was in a movie, but I knew this was actually what it looked like here, cobblestone streets and all.  This was also my first chance to try the Trdelnik, a traditional cinnamon/sugar (+ other tasty bits like chocolate upon request!) pastry that is wrapped around and cooked on a wooden spit. I was not disappointed!

The "Tim Burton" castle, as it was dubbed

Gateway to the Charles Bridge,
over the Vltava River

View of the Prague Castle from the Charles Bridge

View of Prague

Cathedral at the Prague Castle

The conference ended on Friday, and the conclusion was dinner at a local restaurant.  A map was provided, but it took 30 minutes and running into a friend I met previously (who was also lost) to finally find it.  It was in the basement of one of the municipal buildings, and there were almost 0 signs for it. The streets are also terrible to navigate, as they seem to belong in Willy Wonka's factory - just when you think you know which direction you're headed, you end up somewhere else.

Conference dinner

It was fortuitous running into Suzanne at the dinner, because we soon learned that both of us had late flights the next day, and coordinated a trip on Saturday to see more of Prague than either of us had time for earlier. I was really glad she let me tag along because she knew of some places to see that I wasn't aware of, including the old castle where some famous composers are buried, and Emauzy which is a 16th century church with original paintings on the walls.  We also stopped at a riverside farmers market and got some strawberries to munch on as well as another local restaurant, this one cafeteria-style.

Cathedral at the Old Castle

Farmer's market

Emauzy

Original painting

One of the hallways in Emauzy

More meat and potatoes

I am so thankful for my NSF funding which allowed me to finally attend this meeting.  I had a great time meeting new people, learning about new science, and exploring a new part of Europe I had never been to.  However, jet lag has not treated me well and now I will spend a few days recovering...

Another trip to Trinidad

Scarlet Ibis returning from Venezuela

I have just returned from Trinidad (the country, not the city in Colorado) with Cameron Ghalambor, Craig Marshall (a new PhD student), and Travis Klee (a new Bachelor of Science). Unfortunately, we didn't have internet so I couldn't post about the trip as it happened, so instead, I will have try my best to recall the details and events.

Cameron and Craig arrived in Trinidad on June 10, after almost missing their flight from Denver, being delayed in Houston for about 4 hours, and arriving in Trinidad after the rental car place had closed. The next day, the plan was for them to obtain permits, scope out field sites, and purchase some groceries before Travis and I arrived in the evening. They were mostly successful except in regards to the permits (they arrived at the permit office at 3:40 only to be told they closed at 4 and were currently closed, welcome to "Trini time"). At least they managed to track down a rental car by the time Travis and I arrived at 10:30. We also had delays in Houston because the pilot couldn't get one of the engines to start (kind of a big deal!), but we did manage to get upgraded to the front bulkhead row and got free TV. Other than the delay, we made it to the field station with relatively little trouble.

The Reznick guppy facility near Arima, Trinidad

The plan for collections was to get three guppy species from a range of salinities for Cameron and Craig (Poecilia vivipara brackish, P. picta brackish, P. picta fresh, P. reticulata fresh) and 2 high predation/low predation pairs of P. reticulata for me. We started by collecting the brackish fish at a boat dock in the Caroni Swamp on the western side of the island.  We were very successful at collecting fish, but the vivipara turned out to be more difficult to catch with our butterfly nets than we anticipated.  We also tried seining but our net was small and we didn't catch any guppies (though we did catch other things, like eel/leptocephalus larvae). We also managed to find a site close to the boat ramp that was freshwater and were able to sample the freshwater picta. They were densely concentrated under the vegetation along the shore, which made sampling go very quickly. This is ideal since we don't like keeping the guppies in the transport bottles longer than necessary.

The boat canal in the Caroni Swamp

Cameron (left) and Craig (right) catching guppies

Cameron sorting guppies into bottles to be
transported back to the lab

The swamp is about an hour drive from the lab, so unfortunately we had some mortality when we got back.  But being scientists, we took this as a learning opportunity to try to verify the species we caught and distinguish vivipara and picta females.  Because guppies are small, sometimes it is easier to tell these differences on a fish that isn't swimming around.

Craig and Cameron sorting guppies

Examples of some of the fish from the Caroni swamp.
Male picta were obvious and are grouped on the right.
The middle column is most likely picta females, with the
exception of the first two, which are viviparafemale and male.

The next day we headed to the north slope of the mountain range to collect some freshwater reticulata from HP and LP sites.  We stopped quickly at one of the beaches along the Caribbean Sea then headed to our sites on the Yarra River. We sampled the HP site first, and saw lots of other cool fauna, including a large cichlid guarding its young, shrimp, and schools of characins (look like large tetras). Then we headed back up the Blanchisseuse Road (not for those with sensitive stomachs) to the Yarra LP site, which took some exploring to find the dirt road leading down to the river.  When we got there, we were greeted by a group of about 5 guys with a dead goat tied to a tree that was in the process of being butchered.  I couldn't make this up if I tried. They were very cordial though and had no problem with us collecting guppies upstream from them.  They did try to help by telling Cameron about some "big fish" near where they were working, and when Cameron asked if there was blood in the water, the response was "I just killed a goat". We did not sample there.

Las Cuevas Beach

Sampling the Yarra HP site

Sampling the Yarra LP site

We were very fortunate in that the rain held out most of the time we were sampling, so the next day we headed to the Aripo River on the south slope, which drains into the Caroni Swamp eventually.  We sampled a LP site that is also referred to as the Naranjo River.  Though this site has traditionally been sampled extensively, we found a healthy population. However, we did make sure to only remove a small number of individuals from each pool. We found a lot of killifish (Rivulus/Anablepsoides hartii) at this site as well, which only prey on juvenile guppies and are considered a minor predator. On the way home, we also stopped by the site of the Endler LP introduction about 40 years ago.  Again we found a healthy population that could potentially be a site for future studies.

The Naranjo/Aripo River LP site

Looking upstream

Killifish, Rivulus/Anablepsoides hartii back at the lab

After taking the fish back to the lab, we headed back out to the Caroni River again, this time further upstream so I could catch some freshwater reticulata from an HP site to pair with the Aripo LP site. We checked a site that I had sampled the last time we were in Trinidad.  However, that time was in the wet season and the river was difficult to sample so we were left with sampling one of the drainage ditches nearby.  These fish ended up being very unhealthy so not many survived when we returned to Colorado.  This time, however, we were able to sample the river.  The population wasn't as high density as the one near the swamp that we sampled a few days previously, so it ended up being the site that took the longest amount of time to finish, but it was still only about an hour. We had to be quick because just upstream someone was cutting down bamboo stands and were getting closer and closer to us and we didn't want them to fall on us.

Cameron looking for guppies in the Caroni River

Cameron and Craig also needed some freshwater reticulata since their previous freshwater site ended up being about 90% picta, so we went searching for a nearby site they could use.  We ended up near a road crossing in a popular spot for "liming" (aka, partying). Once we located a good vegetation patch with lots of adults hiding in it, it only took a few tries to get all the guppies we needed.  However, in the meantime, Cameron managed to find a fire ant colony that apparently he was sitting on.  He was a trooper because he brushed them off and continued sampling guppies. At this point, we had now collected everything we were targeting, which is pretty good for only 3 days!

Sorting guppies on the Caroni River (post fire ant attack)

Travis and some guppies caught at the Caroni HP sites

After a hard day of exploring, collecting, and fire ant attacks, we decided not to cook dinner, but to instead go out for some Trini street food called doubles.  They're sort of like a taco, with two pieces of bread overlapped on the bottom, then doused with a chickpea sauce and potentially other sauces.  If you're brave, you can ask for "plenty peppa".  I was not brave but the guys were. Each doubles stand is unique and we found some that serve them with other ingredients like ground chicken or a mango sauce.

Eating doubles

Looks like what you might
see in a baby diaper

Cameron had to head back to Colorado the next morning but the rest of us stayed another 2 days.  We used this time to try to catch some pike cichlids (Crenicichla frenata).  These fish are piscivorous and are one of the main predators of guppies in the HP sites.  We needed to catch a few to bring back to Colorado so we can use them for a "predator" treatment in the flow-through systems. We were told the best way to catch these is using hook and line with guppies as bait, so we headed to a few HP stream sites to see what we could do.

The morning started with a downpour and was the official start of the rainy season. We weren't able to get out for the first few hours, but once it cleared up we headed out to the El Cedro River.  We got (vague) directions to the site from a few people, with the warning "DO NOT go to the dump".  Apparently this is where very desperate people live and will attack people looking for anything they think will be valuable to them.  We thought we were going the right way, then saw a man in an orange vest that looked like he was checking in cars at a gate.  Immediately I realized it was the dump, so we had to turn around quickly and head back.  We eventually found the river in what looked like someone's driveway, but after poking around for about 20 minutes, we didn't see much of anything, including guppies.

The El Cedro River

Next, we headed to an Aripo HP site to try again for pikes.  We had to do a little more exploring to find the trail leading down to the river, but eventually found it.  It was kind of fun to actually do more of a hike to get to a site, rather than just pulling over on the side of the road.  This was the first site where I expected to see a fer-de-lance or bushmaster, the two venomous snakes found on the island, but we didn't see any. I guess that's actually a good thing. The river was beautiful when we got to it, and we had a clear view of the waterfall separating the HP and LP sites (predators have a difficult time scaling these, but somehow guppies and killifish can do it). Not long after we arrived at the site, we were overtaken by some kind of flying insect.  They didn't bite, but it was annoying because it made it difficult to concentrate on trying to fish.  I even had one fly into my eye.  When we got back to the house that evening, we learned that the rain earlier in the morning caused the termites to emerge.  We did see a lot of other fish at the site, mostly the larger Characins, but were unsuccessful at catching a pike.

Aripo River and waterfall

The rains brought out a lot of amphibians, and we decided that evening to go on a herp walk to see what we could find.  We probably saw at least 5 species, including ones that were inflated floating in a puddle.  We also saw some cane toads and some other frogs that we weren't sure exactly what they were.

The next (and last) day we tried for pikes again in the Oropuche River.  In contrast to all the other sites we had been to, this river drains to the Niriva Swamp in the east. We tried for a while and were unsuccessful at catching anything using our guppy bait.  This site is also a popular liming spot, so one of the locals came over to help us.  He suggested using some crackers (which he donated to us) to draw the small fish in, and the big fish will follow.  While this seemed like a great suggestion, the river was flowing a little too fast for this to work well.  However, Travis managed to figure out a way to bait the smaller characins and scoop them up with one of the butterfly nets.  Then we were able to use these for bait, which were much bigger than the guppies. After about 2 hours of trying, we had several bites, but were unsuccessful at catching anything. Then we met another local who actually collects fish and works for the zoo in Port of Spain.  He told us they catch them using a seine that spans the width of the river. We didn't have one that big, but we did try walking upstream a bit to find one of the spots he suggested.  We never did catch any pikes.  Fortunately, one of the other researchers went out to El Cedro for killifish that night and was able to net one for us while it was sleeping, so we were able to bring one back in the end.

Travis and Craig fishing for Pike

Me fishing for pike

Marianne, the pike cichlid

We wanted to make sure we did one thing for fun while we were in Trinidad, so the last evening we took a boat tour of the Caroni Swamp to watch the scarlet ibis return to their roost after foraging in Venezuela for the day. We saw lots of other wildlife too!  I'm really glad we had a chance to get out in the swamp a bit.

Me, Travis, and Craig

Four-eyed fish, Anableps

Tree boa

Silky anteater

Scarlet ibis

So in all, I would say it was a very productive trip.  Only about 5% of our fish died on the flight back to Colorado, and all are looking healthy and happy in the lab. Now I have no excuses, time to collect some data!