Behavioral and TRPA1 Heat Sensitivities in Three Sympatric Cuban Anolis Lizards


I would like to introduce our recently published paper on Comparison of Behavioral and TRPA1 heat sensitivities in Cuban Anolis lizards. In Cuba, three sympatric species of Anolis lizards (Anolis allogus, A. homolechis, and A. sagrei) inhabit different thermal microhabitats (above). Different thermal habitats, that is shade, edges of forests and cleared forests, are occupied by A. allogus, A. homolechis and A. sagrei, respectively. Anolis allogus is non-heliothermic, while A. homolechis and A. sagrei are heliothermic species. Our previous study found that these species showed distinct gene expression patterns in response to temperature stimuli, suggesting the genetically distinct thermal physiology among species (Akashi et al. 2016. Mol.Ecol.).

For lizards, heat avoidance behavior is crucial for limiting their body temperatures within thermally safe margins. We predict that the temperature that elicits heat avoidance behavior would differ between these three Anolis species, and the differences might be related to different heat sensors among the species. Organisms perceive various temperatures via biological temperature sensors, such as thermosensitive transient receptor potential ion channels (thermo-TRPs). Among known thermo-TRPs, transient receptor potential ion channel ankyrin 1 (TRPA1) in non-mammalian species has been reportedly heat sensitive (Saito et al. 2012).

In our paper, we first conducted behavioral experiments to analyze the temperatures at which the three Anolis species escape from heat source (i.e., hotplate; Fig. 1) to examine whether the Anolis species inhabiting locally distinct thermal habitats diverge their thermal tolerances.

Then, for each of the three species, we isolated cDNA encoding of TRPA1, and performed electrophysiological analysis to quantify activation temperature of Anolis TRPA1. We found that temperatures triggering behavioral and TRPA1 responses were significantly lower for the shade-dwelling, non-heliothermic species (A. allogus) than for sun-dwelling heliothermic species (A. homolechis and A. sagrei).

The ambient temperature of shade habitats where A. allogus occurs stays relatively cool compared to that of open habitats where A. homolechis and A. sagrei occur and bask. The high temperature thresholds of A. homolechis and A. sagrei may reflect their heat tolerances that would benefit these species to inhabit the open habitats.

Akashi, H., S. Saito, A. Cádiz , T. Makino, M .Tominaga, M. Kawata. (2018) Comparisons of behavioral and TRPA1 heat sensitivities in three sympatric Cuban Anolis lizards. Molecular Ecology  https://doi.org/10.1111/mec.14572

Anoles versus Geckos: The Ultimate Showdown

Two green lizards in Miami, one of each variety.

Two green lizards in Miami, one of each variety.

History is rich with great rivalries; David versus Goliath, Red Sox versus Yankees, Alien versus Predator, but one of the greatest match ups of our time is anole lizards versus gecko lizards. For readers of this blog that are unfamiliar, for which I assume there are few, geckos and anoles are well matched competitors because of their morphological and ecological similarities. Geckos (infraorder Gekkota) are the earliest branch on the squamate tree (sister to all other lizards and snakes) with over 1500 species around the globe, whereas anoles (genus Anolis) appeared roughly 150 million year after the origin of geckos (nested within the Iguania infraorder). The roughly 400 species of anoles can be found primarily in Central and South America. Geckos and anoles both independently evolved very similar hairy adhesive toe pads that help them adhere to and navigate vertical and inverted surfaces. While anoles can likely trace their toe pads to a single origin (and one loss in A. onca), toe pads likely arose and were lost multiple times within Gekkota, although we are still sorting out the exact details (Gamble et al., 2017). Nearly all anoles are arboreal and diurnal, with only a handful of terrestrial or rock dwelling species. Conversely, geckos can be found thriving in arboreal as well as rocky and terrestrial microhabitats day and night.

While anoles tend to get all of the attention from evolutionary ecologists, with decades of amazing research quantifying their habitat use in the Caribbean, geckos are actually older, with more ecological and morphological diversity. As my prior PhD advisor Luke Harmon can surely confirm, I have been interested in knowing how or if insights from Caribbean anole ecomorphology can be applied to geckos. How similar is the evolution and diversification of geckos and anoles? Do geckos partition their habitat along similar dimensions as Caribbean anoles?

In this post, I’d like to share some of my previous work comparing and contrasting gecko and anole diversification and habitat use and then solicit information and opinions from the anole community for an upcoming field trip in which we will be looking at habitat use of sympatric introduced geckos and anoles.

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Fig 1. Our reconstruction of gecko (blue) and anole (green) ancestral toe pad performance based on our best fitting weak OU model of trait evolution. Horizontal bars below the X-axis represent the region in which we constrained the origin of toe pads for each clade. Detachment angle (y-axis) represents our measure toe pad performance (the maximum ratio of adhesion and friction a species can generate). The generation of more adhesion for a given amount of friction results in a higher detachment angle. Shaded bands represent our estimated OU optimum value for each clade. Figure modified from Hagey et al. (2017b).

In 2017, we had two great papers come out investigating the diversification of toe pad adhesive performance in geckos and anoles, and the ecomorphology of Queensland geckos. In our diversification paper (Hagey et al., 2017b), we found that while geckos are an older and larger group than anoles, their toe pad performance does not appear to be evolving towards a single evolutionary optimum. Instead, we found that Brownian motion with a trend (or a very weak Ornstein-Uhlenbeck model) best modeled our data, suggesting geckos have been slowly evolving more and more diverse performance capabilities since their origin approximately 200 million years ago (Fig 1). These results assume a single evolutionary origin of Gekkota toe pads, which was supported by our ancestral state reconstructions, but ancestral state reconstructions are far from a perfect way to infer the history of a trait. And so for now, the true history of the gecko toe pad’s origin(s) remains a ‘sticky’ issue. Conversely, adhesive performance in anoles appears to be pinned to a single optima in which anoles quickly reached after their split from their padless sister group (i.e. a strong Ornstein-Uhlenbeck model, Fig 1).

Given these results and the fact that geckos are such a morphologically diverse group, living on multiple continents in many different microhabitats, our results suggest the adhesive performance of geckos may be tracking multiple optima, and when pad-bearing geckos are considered together as a single large group, could produce the general drifting pattern we observed when we assume their ancestor started without little to very poor adhesive capabilities. On the flip side, we can imagine multiple reasons why anoles appear to be limited in their toe pad performance. Perhaps anoles lack the genetic diversity to produce more variable toe pads or they are mechanically or developmentally constrained to a limited area of performance space. Alternatively, since anoles are nearly all arboreal and diurnal in new world tropical environments, it is possible that they are all succeeding in the same adaptive zone and there isn’t the evolutionary pressure or opportunity to evolve more diverse performance capabilities. Closer studies of the adhesive performance capabilities of the few anoles species that have branched out from arboreal microhabitats, such as the rock dwelling aquatic species would be really interesting!

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Fig 2. Our gecko and anole residual limb length calculations suggest geckos (grey triangles) generally have shorter limbs then anoles (black circles). Figure modified from Hagey et al. (2017a).

In our second paper from 2017 (Hagey et al., 2017a), we quantified microhabitat use and limb lengths of geckos across Queensland, Australia and compared these patterns to those known from Caribbean anoles. We found some interesting differences and similarities. Our first result arose as we tried to calculate residual limb lengths and realized that geckos, as a group, have shorter limbs than anoles, which resulted in us calculating residual limb lengths for geckos and anoles separately (Fig 2). We then compared microhabitat use and limb length patterns and found that Strophurus geckos may be similar to grass-bush anoles. Both groups have long limbs for their body lengths and use narrow perches close to the ground. We also found other general similarities such as large bodied canopy dwelling crown-giant anoles and large bodied canopy dwelling Pseudothecadactylus geckos. Unfortunately, we didn’t focus on sympatric Australian geckos and so we couldn’t make direct habitat partitioning comparisons to anoles. We hope to fix that in our next project and would really love to hear from you, the anole community.

Later this spring, I am planning a fieldtrip with John Phillips and Eben Gering, both fellow researchers here at Michigan State University, to Hawai’i (Kaua’i and O’ahu) to investigate habitat partitioning of invasive geckos and anoles, specifically A. carolinensis, A. sagrei, and Phelsuma laticauda. Jonathan Losos one claimed that Phelsuma are honorary anoles! These three species are all diurnal, arboreal, have adhesive toe pads, and are commonly seen in Hawai’i and so we expect them to be competing for perch space. This has been on some of the greatest anole minds since at least 2011 with Jonathan wondering which group would win when the two clades collide in the Pacific. Previous studies of anole ecomorphs across the Greater Antilles and invasive A. sageri in the southeastern US give us a good expectation of how the trunk-crown A. carolinensis and the trunk dwelling A. sagrei should interact and partition their arboreal microhabitat, with A. sagrei pushing A. carolinensis up the trunk. The wild card is P. laticauda. There hasn’t been much microhabitat use work done with Malagasy geckos, and definitely nothing compared to the extensive work with Caribbean anoles. As a result, I don’t know much about exactly what part of the arboreal environment P. laticauda uses in its natural range or how it will fit in with its new pad-bearing brethren in Hawai’i. The best information we have to my knowledge is a study of other arboreal Phelsuma by Luke Harmon in Mauritius (Harmon et al., 2007). He found that while the Phelsuma geckos of Mauritius also partition their arboreal habitat by perch height and somewhat by diameter, they also partition by palm-like or non-palm-like perches. I’m not aware of any anole observations suggesting a palm/non-palm axis of partitioning and so this may be a novel axis that P. laticauda is using in Hawai’i to live in amongst the anoles.

Anoles, geckos, and Hawai’i have come up repeatedly here on Anole Annals

Reproductive Biology of Introduced Green Anoles in Hawaii

JMIH 2016: Anolis vs. Phelsuma in Hawaii

Amazing Green Anole Battle In Hawaii

More On Anoles And Day Geckos In Hawaii

Anoles And Banana Flowers In Hawaii

Fighting Hawaiian Anoles

Brown Anoles on Hawaii and Battle of the Intercontinental Convergents

Many Hawaiians Don’t Like Brown Anoles

SICB 2018: Unraveling Natural and Human-Mediated Founder Events in Anolis carolinensis

Factors Restricting Range Expansion for the Invasive Green Anole Anolis carolinensis on Okinawa Island, Japan

Anole Watercolor Available on Etsy

A Failed Anole Predation Attempt

This Is Not A Madagascan Day Gecko

Battle of the Diurnal, Arboreal Exotics in Florida (the Anole Loses)

Strange perch mate

Green Anole Mayhem

and so we know folks have been thinking about these species and specifically this invasive set of species for a while. We are especially excited to see Amber Wright’s research suggesting P. laticauda was perching above A. carolinensis in her enclosures. We want to know what the anole community has to say. We also don’t want to duplicate or intrude on any projects that are already under way.. If this is something you’ve already started, or started to wonder about… let us know! We would love to collaborate, partitioning interesting questions, if there are already labs working in this arena. We would also be grateful for suggestions, field site recommendations, or relevant publications we may have missed.

 

Herp Review Bonanza: Green Anoles Mating with Browns, Crested Anole Cannibalism, Brown Anoles Eating a Snake, and Communal Green Anole Nests!

From article by Sater and Smith in March 2018 Herp. Review.

From article by Sater and Smith in March 2018 Herp. Review.

The March 2018 issue of Herpetological Review is chockful of fascinating Natural History Notes about anoles. Highlights: A male carolinensis mating with a female sagrei (we’ve seen that before!), a cristatellus eating a smaller member of the same species, a sagrei eating an anole, and communal nesting in green anoles. You can read all these stories and more, now that Natural History Notes are open access and downloadable! Click on volume 49(1), Natural History Notes.

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Miami Exotic Lizard Safari

A very ambitious brown anole

A very ambitious brown anole

I arrived a day early for the 2018 Anolis Symposium. When it became clear I was not needed to help get things ready, I did what any red-blooded anolologist would do: I headed off for All-America Park, the hottest of hotspots for Miami anoles.

And what a day it was. Two minutes after leaving my hotel, I saw what I’m pretty sure was an Ameiva, though I didn’t get a good look. Then red-headed agamas underneath the monorail on the Dixie Highway.

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Plenty o’ iguanas. How many can you find?

How many green iguanas can you find?

How many green iguanas can you find?

And curly-tailed lizards!

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Along the way, I also saw three introduced anoles: (A. sagrei, A. cristatellus and A. distichus).

Anolis distichus

Anolis distichus

Finally, I got to All-America Park and immediately met five-term South Miami mayor Phil Stoddard, who doubles as a crackerjack neuroethologist at Florida International University, and is a great naturalist to boot. We walked around the park looking for Anolis garmani, the Jamaican giant anole, but without success. There were plenty of other anoles at the Park, all the same ones I’d already seen, but also a knight anole and 13 green anoles–they definitely are doing just fine despite all the invasives. The lizard search was conducted to a soundtrack of screeching peacocks and the occasional flock of parrots flying back. I love Miami! Sadly, no Jamaican giant anoles, A. garmani. That’ll have to wait for a return visit.

Anolis equestris at All-America Park

Anolis equestris at All-America Park

Anolis Symposium Tomorrow!!!

Fairchild Botanical Gardens, site of the 2018 Anolis Symposium

It’s not to late to hop on a plane and get to beautiful Fairchilds Botanical Garden! Abstracts are posted. Here’s the schedule:

9:00 – 9:30 James Stroud Florida International University
Introduction and Welcome

9:30 – 9:45 Michele A. Johnson Trinity University
Physiological mechanisms underlying behavioral convergence in Caribbean anoles

9:45 – 10:00 Tony Gamble Marquette University
Anolis sex chromosomes, past, present, and future

10:00 – 10:15 Rosario Castañeda Universidad Icesi
When did anoles diverge? An analysis of multiple dating strategies

10:15 – 10:30 Colin Donihue Harvard University
Hurricane-induced adaptive shifts in the morphology of an island lizard

10:30 – 10:45 Leo J. Fleishman Union College
Why are there so many yellow dewlaps?

10:45 – 11:30 Coffee Break

11:30 – 11:45 Graham Reynolds University of North Carolina Asheville
Genetic and Morphometric Diversification in the Brown Anole Suggest Early Pathways of Anole Colonization and Evolution in the Caribbean

11:45 – 12:00 Nathalie Feiner Lund University
Transposable elements, Hox gene clusters and genome evolution– How special are Anolis lizards?

12:00 – 12:15 Thomas J. Sanger Loyola University Chicago
The Mechanisms of Thermal Stress Induced Craniofacial Malformation in Lizards Developmental biology

12:15 – 12:30 Sozos N. Michaelides University of Rhode Island
Invasion history of four Anolis lizard species introduced to Bermuda Invasion biology

12:30 – 14:00 Lunch

14:00 – 14:15 Kristin M. Winchell University of Massachusetts Boston
Performance Consequences of Urban Morphological Shifts

14:15 – 14:30 Kenro Kusumi Arizona State University
Comparative Genomics Reveals Accelerated Evolution in Conserved Pathways during Anolis Diversification

14:30 – 14:45 Sean Giery University of Connecticut
Some thoughts on the trophic ecology of Anolis lizards

14:45 – 15:00 D. Luke Mahler University of Toronto
Land use and the restructuring of anole communities across an elevational gradient

15:00 – 15:45 Coffee Break

15:45 – 16:00 Ivan Prates Smithsonian Museum of Natural History
Genomic signatures of adaptation associated with a history of range expansions in South American anoles

16:00 – 16:15 Oriol Lapiedra Harvard University / CREAF
Predator-induced natural selection in behavior Behaviour

16:15 – 16:30 Caitlin C. Mothes University of Miami
Using South Florida’s exotic lizard community to evaluate the use of ecological niche models in predicting biotic invasions

16:30 – 17:00 Neil Losin Day’s Edge Productions
The Lizard’s Tale and Anole Annals v2.0: An enhanced platform for Anolis outreach

17:00
Social & Poster Session

Saturday, 17 March 2018
9:30 – 9:45 Douglas B. Menke University of Georgia
Genome editing methods for the production of genetically modified anoles

9:45 – 10:00 Sarin Tiatragul Auburn University
A shady way to beat the Miami heat

10:00 – 10:15 Joanna O. Palade Arizona State University
Anolis carolinensis satellite cells have expanded musculoskeletal potential

10:15 – 10:30 Gregory C. Mayer University of Wisconsin-
Parkside Using archival DNA to elucidate anole phylogeny Systematics and/or taxonomy

10:30 – 10:45 Liam J. Revell Universidad del Rosario and UMass Boston
Can we detect differences in the rate of discrete character evolution between clades of anoles?

10:45 – 11:30 Coffee Break

11:30 – 11:45 Amber N. Wright University of Hawaii
Predicting the outcome of species interactions in a novel species assemblage: Anolis vs. Phelsuma in Hawaii

11:45 – 12:00 Andrew C. Battles University of Rhode Island
The other Miami Heat: Urban areas alter thermal biology and influence persistence and spread of two invasive Anolis species.

12:00 – 12:15 Nathan W. Turnbough I
Covariation in arthropod community composition and dominant anole identity on dredge spoils islands in Florida

12:15 – 12:30 Cindy Xu Arizona State University
Tail Regeneration in Anole Lizards: Insights from Comparative Genomic Analysis and Reformation of the Peripheral Motor Nervous System

12:30 – 14:00 Lunch

14:00 – 14:15 Michael L. Logan Smithsonian Tropical Research Institute
Using experimental islands to explore evolutionary dynamics under climate change Thermal biology, ecology, or evolution

14:15 – 14:30 Christine Rose-Smyth Verdant Isle Orchids
Role of a sweet-toothed anole in orchid pollination Species interactions

14:30 – 14:45 Christopher J. Thawley University of Rhode Island
Let There Be Light: Widespread Use of Human-Produced Light at Night by Anoles and Its Consequences

14:45 – 15:00 Sean Doody USF St. Petersburg Environmentally Cued Hatching in Anoles Behaviour

15:00 – 15:45 Coffee Break

15:45 – 16:00 Winter A. Beckles University of Miami Signal divergence and habitat partitioning among non-native bark anoles in South Florida

16:00 – 16:15 Stephanie L. Clements University of Miami
Non-native species dominate herpetofaunal community composition in both native and non-native habitat patches in Miami-Dade County

16:15 – 16:30 Zachary A. Chejanovski University of Rhode Island
Predators influence prey body size variation in an urban landscape

16:30 – 16:45 Joshua M. Hall Auburn University
Does season-dependent reproductive value of offspring drive the evolution of life-history traits in Anolis lizards?

16:45 – 17:00 Jonathan Losos Washington University in St. Louis
Concluding Remarks

A New View on Anole Territoriality and Social Structure

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One of our marked lizards for this study. Photo by Jon Suh.

Ambika Kamath has written a synthesis of her two recent papers in territoriality in anoles. It appeared on March 12, 2018 on her blog, Behavioral Ecology, Natural History, Science and Society and is reprinted below:

In my major Ph.D. project, I questioned the idea that territoriality is a good or useful description of Anolis lizards’ mating systems. When I began working on this question, I planned to primarily use an empirical approach, measuring the movement patterns and mating patterns of a population of Anolis sagrei in a way that didn’t depend on territoriality. But anticipating future criticism, I realised that because I’d be working in one population of one species, my empirical work could readily and reasonably be dismissed as an aberration without a broader foundation on which to place it.

This realization led to the historical review in which my Ph.D. advisor Jonathan Losos and I examined the history of research on Anolis territoriality. I’ve written about this historical research quite a bit before, but haven’t said much about the empirical work, leaving the two complementary halves of this project unintegrated. That’s partly been because the empirical work wasn’t published until recently. But it’s also because in contextualizing the problem tackled by the empirical paper, I have to basically recount the whole of the historical review. There really hasn’t been room to talk about both in a single venue, and there still isn’t, but I’m going to tell you a bit more about the empirical paper to balance things out. You’ve heard a little about it before–I wrote field notes about one of the males in this study (interesting addendum: U131 fathered none of the offspring of the females he encountered!) and about a tiny survey of green anoles that we conducted concurrently.

The empirical paper is now published, in the Proceedings of the Royal Society B! Here’s an awesome press release about the study from UCSB that will give you the gist of it, but in short what we did was:

  • Catch and mark almost every lizard we saw, and then measure the spatial locations of as many lizards as we could by repeatedly surveying as big an area as we could.
  • Make a map of all the trees within our sampling area.
  • Measure the body size and estimate the population-level growth rate of males
  • Collect a subset of the females, bring them into the lab, and collect the DNA of their offspring.
  • Devise a mathematical approach to estimating encounters between males and females from data on their spatial locations. Combined this with the growth-rate estimate to calculate the size of males at their encounters with females.
  • Use DNA sequencing to figure out the likely fathers of the females’ offspring; we leaned on the estimates of male-female encounters to do so.
  • Use a clever and (I think!) pretty original approach to quantifying sexual selection on body size and movement patterns by comparing the traits of males that encountered females to the traits of the subset of those males that actually fathered offspring.

In sum what we found was that male and female movement patterns spanned larger areas and were more dynamic than many of us had previously imagined, that females encounter multiple potential mates, that at least 60% and possibly up to 80% of females  mate with multiple males, and that sexual selection acts on male body size as well as males’ spatial extent and the timing of male-female encounters. I’ll let you read the press release and the paper itself to learn more about what we found (here it is on BioRxiv, essentially the same paper but freely accessible)!

Viewed together, I hope the historical and empirical papers make a convincing case that we’ve been looking at Anolis mating systems in a limited way for a long time, and that other, newer ways of quantifying mating systems in ways that don’t depend on territoriality can yield both interesting and sensible results. I see this work as opening up an arena of questions, both in Anolis and in other taxa where mating systems have been described in a static way for a long period of time.

I’m very proud of this paper. I remember a phase of grad school when I found it impossible to convince people that this work would turn out interesting, or maybe it was just that my own self-doubt prevented me from seeing others’ interest and support for this research. It remains true that this is one study of one population of one species, and it may well be that I turn out to be all wrong. Perhaps new explorations of Anolis mating systems will eventually lead us back to territoriality. But even if that’s the case, I feel confident that, thanks to this work, we’ll be able to approach that or any description of Anolis mating systems with clearer, more skeptical, and more discerning eyes.

This won’t be the last you’ll be hearing from me on this subject of lizard mating systems; for one, there are responses to our historical review that are in the process of being published, and we’ll have a chance to respond to them. I’m very excited to engage in an actual scientific dispute, and will do my best to do so respectfully and productively, especially since I have on-the-record views about what makes such disputes annoying. But in terms of research, I seem to be heading in other directions, which I think will be related to this work but maybe not directly. So I wanted to make sure that I put down here, all in one place, what I see this project as and what I hope it will achieve. Let me know what you think!

A Case of Matestaken Identity: Hybrid Mating between Crested and Brown Anoles!

Somebody needs to work on their anole species identification skills.

Somebody needs to work on their anole species identification skills.

Breeding season is heating up for anoles in Miami, and at least one male crested anole (A. cristatellus) is a little…confused. While collecting some baseline data for my post-doc work looking at impacts of artificial light at night (ALAN) on brown and crested anoles, I noticed a commotion on a nearby cycad. Upon closer inspection, I realized that a male crested anole was pursuing and subsequently mating with a female brown anole (A. sagrei) who was decidedly unhappy about the situation.

In case you’re wondering about the colorful jewelry at the base of their tails, both of the anoles in the photo/video are bead-tagged to allow me to reidentify them from a distance. The copulation here lasted 3-4 minutes a portion of which I managed to capture on video.

While previous reports on AA have documented coupling between A. carolinensis and A. sagrei, I haven’t seen any reports of interspecific mating between A. cristatellus and A. sagrei. Has anyone else observed this phenomenon? The two species do encounter each other quite frequently in the Miami area, so this might not be a rare occurrence. Hybridization seems unlikely given the divergence between these two species, but you never know!

Congratulations Dr. Kristin Winchell!

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Yesterday afternoon Kristin Winchell successfully defended her Ph.D. dissertation at UMass Boston…during a nor’easter! The university was officially closed due to the inclement weather (I myself got stuck in a stairwell for several minutes because I didn’t have card swipe access), but that didn’t stop Kristin from delivering a fabulous talk about her work on urban evolution in anoles.

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Congratulations Kristin!! And can you believe this cake?!?!

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James T. Stroud, PhD!

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The latest Dr. Anole…but not for long! Stroud wowed a packed house at Florida, regaling the audience with four chapters of research, two of which are already published in Annual Review of Ecology, Evolution & Systematics and Invasion Biology (2 of the ca. 40 papers he published during his time at FIU. The self-proclaimed highlight of his work? Publishing on T. rex!

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Congratulations, James!

Big Day Tomorrow: Two New Anole Doctors!

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In an event probably unprecedented in anole history, two new anole doctors will emerge tomorrow. After a multi-year incubation, AA stalwarts Kristin Winchell and James Stroud will hatch tomorrow almost simultaneously. James will get the festivities rolling at 11 a.m. eastern time in Miami: 

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Kristin follows shortly thereafter at 1 p.m. in Boston:

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By this time tomorrow, the ranks of Dr. Anolis will be increased by two. Congratulations Kristin and James!

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