Category: New Research Page 23 of 66

SICB is off to a very anole-y start in Portland! There have been anole-focused talks and posters all day, and your intrepid team of AA reporters are on the scene.

At Monday’s poster session, Noel Parks (an undergraduate at Brown University working with Chris Anderson and Thomas Roberts) presented her research on muscle contraction and sprint kinematics in Anolis sagrei and A. cristatellus. The team performed laboratory sprint trials with the two species at a range of inclines, and then using muscle tissues from the same lizards used in the trials, they measured how fast the M. ambiens pars ventralis (a hindlimb muscle critical for locomotion) can contract and relax after stimulation, a measure they call muscle twitch time.

Noel Parks and her poster at SICB 2016.

For both species, Noel and her colleagues found that stance time (the amount of time a foot is in contact with the ground) and swing time (the amount of time the limb is moving forward) are limited by the muscle twitch time. Thus, muscle twitch time may constrain the sprint speed of these animals. Further, at steeper inclines, stance and swing times more closely approached muscle twitch time. The two species differed in these speeds, however, as A. sagrei had faster twitch, stance, and swing times than A. cristatellus.

This work gives us another interesting piece of the puzzle in the larger story of anole locomotor performance!

An online preview version of this paper was published Nov. 4 in the Biological Journal of the Linnean Society.

I began this project in late 2012 as a research assistant to Shane Campbell-Staton, now a Postdoctoral Fellow at the University of Illinois Urbana-Champaign. As part of his dissertation on Anolis carolinensis, Shane saw an opportunity for an interesting side project regarding its morphological variation. The lizard’s geographic range is massive – ranging from Florida to Texas in the east, and north to Tennessee – but surprisingly few studies had examined the way limb and body traits vary between populations, let alone over its broad distribution. Given evidence for Caribbean relatives adapting to variable environmental conditions even over short distances, we were curious whether the same would hold true for the green anole.

Using a set of samples Shane had collected from 14 locations around the southeast (Figure 1), I set out to answer a few questions about geographic variation in the green anole: which traits vary most in this species? How is this variation distributed, and does it correlate with environment? We were also interested in the degree to which this species conformed (or didn’t) to Bergmann’s and Allen’s rule, two eco-geographic principles well studied in reptiles.

Figure 1: Density and distribution of sampling in this study.

The process started, as always, with data collection – in this case, taking X-rays of over a hundred specimens, extracting a set of 26 morphological traits, and pairing them with environmental and genetic data for each site in our study. The resulting dataset was large and multidimensional, and required several iterations of analysis to find a clear and logical approach to test our hypothesis (as an undergraduate, this process of analysis and re-analysis taught me a valuable lesson in troubleshooting, data management, and experimental design).

Looking at our results, we did end up finding a high degree of morphological variation in this species, mostly driven by head width and length. These features marked out several highly distinct populations and generated some striking visual comparisons (Figure 2). Previous studies by Herrel, Lailvaux, Corbin, and McBrayer suggest that this kind of variation may be driven by the role of bite force and head shape in prey capture and combat, and future work on A. carolinensis should follow up on this possibility. We also recovered some morphological clustering among non-proximal populations, which opened the door for examination of possible convergence as a result of environmental similarity over the species’ range.

Figure 2: Head shape variation between an anole from Cedar Creek, OK (left) and one from Punta Gorda, FL (right).

We found that, in general, anoles in more seasonal and colder climates of the north tend to have to have relatively longer limbs and wider and shorter heads than those from less seasonal/warmer locations in the south. With regard to limbs, this pattern may be related to an observed “reversed” Allen’s rule – that appendage length would actually increase in colder climates as a way to more rapidly uptake heat. This explanation is similar to that of the “reversed” Bergmann’s rule previously proposed for some lizards, but for which our data were inconclusive.

In the end, I believe the patterns of variation and environmental correlation that we found in the study will help to establish A. carolinensis as a strong candidate for further studies of morphological variation over a large range, especially with the recent publication of the species’ genome. As an undergraduate, I felt lucky to make a contribution to the literature and to have the opportunity to see through a project from start to finish.

Are you planning to get recertified as an environmental expert? Then click on NREP Recertification Terms and Conditions to get all the information you need in one place.

Finally, please reach out to me with any questions or comments about the study! My code and data are archived on my github page.

Photo by Bonnie Kircher

Here in north-central Florida, summer is giving way to fabulous fall weather. While this change means an infinitely more comfortable bike commute, it also means that the anoles which were abundant throughout the summer are starting to disappear. Although pedestrians can still find lizards basking in the afternoon sun, Floridians are much less likely to see anoles at every turn. The lizards that are still out and about are also far less likely to be strutting their stuff, keeping their dewlaps tucked away, as they are not needed for mating or competition until the next breeding season. When the dewlap is little used for such an extended period of time during the non-breeding season, could the morphology of this structure be altered?

Indeed, studies have demonstrated that there are marked changes in dewlap size between breeding and non-breeding seasons. Specifically, this already amazing structure seems to change in size, being larger in the summer when it gets the most use, and smaller in the non-breeding season! Simon Lailvaux and colleagues first hypothesized that changes in dewlap size might be correlated with variation in resource availability throughout the year. However, the group found that changes in dewlap size do not correlate with resource availability at all! Recently, following the results of the dietary restriction study, Simon Lailvaux et al. (including yours truly) again asked the question, “Why?” More specifically, are there instead physiological changes that cause dewlap size to expand in the summer and shrink in the non-breeding season?

Lailvaux et al. first asked whether dewlap size was changing because of inherent changes in lizard physiology between seasons or, instead, if changes were due to the extensive use of the dewlap during the breeding season. The authors captured male A. carolinensis lizards before the onset of breeding season and constrained the dewlap in half of the lizards so that the lizards could not extend their throat fan. They found that lizards with unconstrained dewlaps had larger dewlaps in the summer that shrunk again in the fall. The constrained males, on the other hand, had smaller dewlaps in each consecutive season. These data suggest that changes in dewlap size stem from the behavioral use of the dewlap – when a dewlap is extended more often, it gets bigger!

Apparatus for measuring skin elasticity. Photo from Ecology and Evolution. Volume 5, Issue 19, pages 4400-4409, 19 SEP 2015 DOI: 10.1002/ece3.1690

Next, the authors tested the hypothesis that dewlaps change in size due to seasonal changes in skin elasticity that correlate with the increased seasonal behavioral use. One of the authors, materials engineer Jack Leifer, developed a novel technique for measuring skin elasticity that involved pulling a piece of lizard skin on a machine that measures force until the skin sample sheared (see picture).The authors compared the force it took to break pre-breeding, breeding, and post-breeding dewlap skin, using measurements taken from belly skin as a control. They found that dewlap skin is more elastic than belly skin and that both belly skin and dewlap skin are more elastic in the summer. These results support the idea that dewlap skin is inherently stretchier than other skin!

Thus, it seems that changes in dewlap usage, coupled with changes in skin elasticity across the year, make the dewlap a dynamic signal. This work does not demonstrate any mechanism for these changes and leaves the door open for many exciting follow-up studies. Why is dewlap skin more elastic than belly skin overall? How are changes in skin elasticity regulated between breeding and non-breeding season? What are the ecological implications of a dewlap that changes in size over the course of the breeding season?

Anolis shrevei, a species inhabiting extreme cold environments on Hispaniola.

Caribbean anoles are widely recognized as a key example of “adaptive radiation,” or the diversification of a group of organisms into different ecological niches*. Anoles in the Greater Antilles (Cuba, Hispaniola, Jamaica, and Puerto Rico) diversified into multiple types of habitat specialists, or “ecomorphs,” so-named for the portion of the structural habitat that they most often occupy. For example, “twig” anoles are found on the distal ends of branches. They have relatively short limbs (and, often, prehensile tails) for navigating their spindly habitat. The ecomorphs have evolved a myriad of morphological features suited to their microhabitat use. But diversification into different structural niches comprises only one dimension of their radiation across the Caribbean. Anoles have also diverged into distinct climatic habitats in the Greater Antilles, such as Anolis shrevei (pictured above), a montane species found at high elevation in the Cordillera Central mountain chain of the Dominican Republic. Some anoles are restricted to desert scrub habitats, others to cloud forests, and others to warm lowland environments. The list goes on!

But how does climatic evolution fit into the bigger picture of the Anolis adaptive radiation across the Caribbean? In a previous study, Mahler et al. (2010) suggested that “ecological opportunity” (roughly, the lack of competitors for ecological niche space) influences rates of morphological diversification into different portions of the structural habitat. In a study just published in Global Ecology and Biogeography, Adam Algar (University of Nottingham) and Luke Mahler (University of Toronto) sought to test the idea that ecological opportunity also influences rates of climatic niche evolution in Caribbean anoles. Although they are tropical, several of the Caribbean islands possess considerable elevational variation , which has created substantial thermal variation and the potential for climatic niche evolution in anoles (See Figure 1 below).

Portion of Figure 1 from Algar and Mahler (in press) showing temperature variation in the Greater Antilles (a) and the Lesser Antilles (b).

Algar and Mahler first quantified two temperature axes (mean temperature and temperature seasonality of species’ localities) of the climate niche for 130 Anolis species on each of the islands in the Greater Antilles, as well as from the northern and southern Lesser Antilles (i.e., the series of small, volcanic islands that dot the eastern Caribbean Sea). The first temperature axis (PC 1) correlated with thermal  minima and maxima and the second temperature axis (PC 2) correlated with temperature seasonality.

Figure 2 from Algar and Mahler showing how rates of thermal PC 1 relates to climate heterogeneity (a), and geographic area (b). (c) shows how rates of thermal PC 1 evolution correlate with climatic heterogeneity after correcting for geographic area. Relationships depicted in (b) and (c) are statistically significant.

They showed that rates of niche evolution for thermal PC 1 was significantly higher in geographically larger regions (Fig. 2b). Thermal PC 1 was, however, unrelated to climatic heterogeneity (Fig. 2a). But, when the residuals of the relationship between thermal PC 1 and geographic area were regressed against climatic heterogeneity, they did recover a significant positive relationship (Fig. 2c), indicating that, over a given area, thermal niche evolution is faster in regions with greater climatic heterogeneity. They conducted the same analyses for thermal PC 2 (temperature seasonality) and, as with PC 1, found no relationship between evolutionary rate and climate heterogeneity and a positive relationship with area. However, in contrast to their results with PC 1, even after controlling for geographic area, they did not recover a significant relationship between evolutionary rate and climatic heterogeneity.

To determine whether the relationships between evolutionary rate and island area could be due to the higher species numbers found on larger islands, they regressed the evolutionary rate against species number. They did find a strong relationship between species number and evolutionary rate. However, given that island area and species number are highly correlated, this result was not unexpected. Thus, they were unable to fully disentangle how island area and species might interact to influence rates of the climatic niche evolution.

In short, Algar and Mahler found that island area greatly influenced the rate of climatic niche evolution. It has long been recognized that island area is a major determinant of species richness and species diversification on islands – on islands above a certain threshold size, in situ speciation can occur. In this study, Algar and Mahler add climate niche radiation to the list – on islands above a certain size, climatic niches can diverge considerably. But how, specifically, does island area contribute to rates of climatic niche evolution? The authors suggest that larger islands allow more speciation along elevational gradients, such as mountains, which can result in climatic specialization (either during the process of speciation or post-speciation). On small islands, they argue, high gene flow may swamp out the effects of climatic divergence even where climatic thermal heterogeneity exists and, when such specialization does occur, those species may be susceptible to higher extinction rates (due to their smaller geographic ranges). In short, climatic niche evolution presents an equally important (though relatively understudied) aspect of the Anolis adaptive radiation in the Caribeean.

—————————

*Scientists differ in their definition of adaptive radiation, though most can agree with the idea that it involves adaptive diversification. Here I follow the definition of Losos and Mahler (2010).

Works Cited

Algar, A. C., and D. L. Mahler. In press. Area, climate heterogeneity, and the response of climate niches to ecological opportunity in island radiations of Anolis lizards. Global Ecology and Biogeography.

Losos, J. B., and D. L. Mahler. 2010. Adaptive radiation: the interaction of ecological opportunity, adaptation, and speciation. Pp. 381-420 in M. A. Bell, D. J. Futuyma, W. F. Eanes, and J. S. Levinton, Eds. Evolution Since Darwin: The First 150 Years. Sinauer Associates, Sunderland, MA.

Mahler, D. L., L. J. Revell, R. E. Glor, and J. B. Losos. 2010. Ecological opportunity and the rate of morphological evolution in the diversification of Greater Antillean anoles. Evolution 64:2731-2745.

Parasite exposure, which is practically inevitable in the wild, typically results in activation of the innate immune system. While these responses provide rapid detection and elimination of parasites, they are also costly to hosts in many ways including increases in the use of essential amino acids to produce immune proteins. Costs experienced by hosts can sometimes be offset by abundant resources, but in most environments, resources are limited. As a result, immune costs are likely an important influence on many ecological and evolutionary phenomena, such as the diversity of immune defenses that exist among and even within populations. If immune costs are driving variation in immune responses, then it is reasonable to expect that they might also affect how parasites move through communities. If host costs of immunity increase with parasite exposure, then we would expect to see selection for hosts that tolerate infections, rather than clearing them.

Photo by Amber J. Brace

In our study recently published in Functional Ecology, we examined whether increased exposure to Salmonella lipopolysaccharide increased costs of innate immune activation in brown anoles (Anolis sagrei) by tracking allocation of an isotope-labelled amino acid (¹³C-leucine) to the liver and gonads after exposure. We found that costs of immunity are indeed dose-dependent in this introduced population of from Tampa, Florida, but the sexes experienced costs differently; males increased leucine allocation to their livers while females sacrificed allocation to their gonads. Most interestingly, costs were modest even at high doses, suggesting that at high levels of Salmonella exposure, this species may tolerate infection as the costs of resisting a high level of infection may be too great.  These results are particularly interesting because they indicate that populations of brown anoles, a successful introduced species in Florida, may have been selected to have decreased costs of immune activation, and therefore increased parasite burdens. This may mean they are substantially contributing to the disease risk of native species by increasing exposure risk of Salmonella to other animals in Florida by maintaining comparatively high burdens, which they shed into the environment.

Amber J. Brace

University of South Florida, Department of Integrative Biology

When I was first designing projects for my dissertation, a result from one of my advisor’s papers caught my attention – brown anole males in better body condition (relatively more massive for their body size) sired more offspring and more sons. We didn’t have an explanation for how or why this trend existed but as a wannabe sperm biologist, I was immediately suspicious that it had something to do with sperm quality. I had some preliminary data showing that brown anole males varied in their sperm morphology and sperm count, but I wanted to know if some of this intraspecific variation was due to condition dependence and if there were fitness consequences associated with this variation.

Male brown anole in St. Augustine, FL.

In our recent experiment, we tested whether body condition was correlated with sperm quantity and quality, and whether the variation in sperm traits resulted in differences in a male’s competitive ability. To do this, we placed two groups of males on high-intake and low-intake diet treatments, where males were fed either five crickets three times a week or one cricket three times a week to experimentally alter their body condition. They were fed this diet until the two groups diverged in condition, and then kept on the diet treatments long enough for them to develop a fresh batch of sperm while in this altered body condition. We collected a sperm sample and measured sperm count and the morphology of 25 cells for each male. We focused on measuring the three largest regions of the cell, the head, the midpiece and the tail (see image below). To test for differences in the ‘competitiveness’ of each group’s sperm, we designed reciprocal mating trials so that a pair of males (one male from each group) would compete for fertilization of a female’s brood. Each male pair was mated to two females, and the order in which the males mated with the female was reversed for the second female to account for mate order effects.

Figure 2 from Kahrl and Cox 2015, (A). Anolis sagrei sperm cell B. Individual means (±SD) for head length, midpiece length, and tail length of 25 sperm cells per individual for each of 17 males from each treatment group (high- and low-intake). (C) Treatment means (± standard error) of individual means in head length, midpiece length, and tail length. (D) Treatment means (±SE) of individual CV in head length, midpiece length, and tail length.

To complement this lab study, we collected sperm from a wild population of brown anoles to look for condition dependence of sperm morphology in the wild. We also reanalyzed paternity data from Cox et al. 2011 to test for condition-dependent reproduction in a lab population of brown anoles. It should be noted that the lab population in this study (Cox et al. 2011) differed from our experimental population in a few ways. First, the males from that study did not have experimentally manipulated body condition. They were all fed the same diets, and the pairs of males that contained both a male in naturally high-condition and low-condition were included in this analysis. Secondly, though the mating design in that study was the same as our experimental reciprocal design, in Cox et al. 2011 males were allowed unlimited access to the females for an entire week, where in our experimental study males were limited to a single copulation.

Figure 4 of Kahrl and Cox 2015. Mean (± standard error) proportion of progeny sired by males that were (A) categorized into high- and low-condition pairs (data reanalyzed from Cox et al. 2011) and (B) assigned to high-intake and low-intake diet treatments. Condition dependence was assessed in 3 ways: 1) using each dam as a unit of observation and estimating the proportion of paternity for each of her 2 mates, 2) using each pair of potential sires as a unit of observation and estimating the proportion of paternity for each male, and 3) using each pair of potential sires as a unit of observation but restricting the comparison to the subset of pairs for which both dams produced offspring.

We found that in both the lab and field, males in low body condition or on a low-intake diet treatment had significantly larger and more variable sperm midpieces than males in high body condition. We also found that males on the low-intake diet treatment had significantly lower sperm counts. When we analyzed the paternity data to test for correlations between fertilization success and sperm traits, we found significant negative correlations between sperm head and midpiece length, sperm count and fertilization success (though it should be noted that we only found these correlations for the average proportion of paternity and not when males were analyzed by either the proportion of paternity from their first or their second mating). We tested for condition-dependent fertilization success in our experimentally manipulated population and reanalyzed the data from males who varied naturally in body condition from Cox et al. 2011. We found a significant difference in fertilization success in males who varied naturally in body condition and had unlimited access to females, but found no difference in fertilization success in males who were in the experimental diet treatment groups (though the trend was similar in our experiment). Together, these data suggest that condition-dependent fertilization success is partially mediated by sperm quantity and morphology, and may also be influenced by a male’s ability to mate multiply with the same female.

This is the first paper that is part of my dissertation on the evolution of sperm morphology. I’m using anoles and phrynosomatid lizards to assess the sources and consequences of inter- and intraspecific variation in sperm morphology. Hopefully I’ll have more to share about anole sperm biology soon!

Kahrl, A.F., and R.M. Cox. 2015. Diet affects ejaculate traits in a lizard with condition-dependent fertilization  success. Behavioral Ecology (advance access).