SICB 2022: Ecological and Genetic Basis of a Sexual Signal

This year at SICB, I had the great opportunity to talk about part of my work as a postdoctoral researcher in the lab of Dr. Michael Logan at the University of Nevada, Reno. In collaboration with John David Curlis (University of Michigan), Christian Cox (Florida International University), W. Owen McMillan (Smithsonian Tropical Research Institute), and Carlos Arias (STRI), we have been studying the Panamanian slender anole Anolis apletophallus, which has a dewlap polymorphism: males either have a solid orange dewlap (solid morph) or a white dewlap with an orange spot (bicolor morph). Preliminary results from John David Curlis’ PhD dissertation research suggests that, in our mainland study population, the frequencies of these morphs change in conjunction with understory light levels—the solid morph is more frequently observed in brighter areas where more light reaches the understory, whereas  the opposite is true for the bicolor dewlap, which is more frequently observed in darker areas of the forest. Thus, it seems possible that selection is maintaining this polymorphism following the predictions of the sensory drive hypothesis, which states that sexual signals should have characteristics that make them the most transmissible given the physical characteristics of the local habitat.

As part of an effort to understand how this trait is evolving in the wild, I set out to understand the genetic basis of this dewlap polymorphism. To do this, my collaborators and I first assembled the full slender anole genome which we then used as a reference for a pooled population sequencing (Pool-Seq) approach using half individuals with solid dewlaps and half individuals with bicolor dewlaps to identify the genomic region underlying this dewlap polymorphism.

Our genome assembly showed pretty good results (Scaffold N50 154,613,287). The Pool-Seq results presented a clear peak of differentiation between solid and bicolor morph groups that corresponded to a region on Scaffold 3. We have a promising candidate gene within this region that may underly the dewlap polymorphism, but will continue to explore these data further to understand the genetic basis of this charismatic trait.

Making the Fancy Feet of Anoles and Geckos

A mourning gecko (Lepidodactylus lugubris) climbing vertically on glass with the help of its impressive toe pads.

I think most people visiting Anole Annals could argue that the adhesive digits of anoles are some of the most fascinating aspects of their biology (or maybe I’m just biased). Digital adhesion is accomplished through toe pads: a collection a broad, modified plantar scales which bear thousands upon thousands of microscopic, hair-like structures (i.e. setae). Through frictional and van der Waals forces, these collections of setae allow toe pad-bearing lizards to easily access vertical surfaces and exploit habitats many lizards cannot. Shockingly, adhesive toe pads have independently evolved several times across lizard evolutionary history (at least 16 times by recent estimates) — once in the common ancestor of anoles, once in a clade of southeast Asian skinks, and 14 times in geckos. Both within and between the different evolutionary origins of toe pads, there is substantial variation in toe pad size, shape, number of scansors/lamellae, and position of the adhesive apparatus.

In our recent study, my collaborators and I took the first steps to characterize how embryonic development is modified to achieve this incredible diversity. Using embryonic material my coauthor Thom Sanger collected as a postdoctoral researcher in Marty Cohn’s lab, in addition to embryonic material I collected over the course of my Ph.D. training in Tony Gamble‘s lab, we aimed to compare embryonic digit development of ancestrally non-padded lizards with that of anoles and padded geckos. We used a model clade approach to broadly sample anoles and geckos, although some species breed more easily in the lab and have more embryological resources than others. All together, we sampled a range of toe pad morphologies in both clades (trunk-ground and trunk-crown Anolis ecomorphs and leaf-toed and basal pads in geckos). To help polarize the developmental changes leading to the origin of toe pads, we also included two ancestrally padless species in our comparisons. After the collection of these diverse embryos, we used scanning electron microscopy (SEM) to characterize scale morphology of the digits throughout embryonic development.

By comparing embryonic material of anoles and geckos, we essentially span the diversity of squamates in a single comparison.

Because of the ~200 million year divergence between anoles and geckos and dramatic differences in adult morphology, we anticipated that we would see stark differences in the developmental origins of toe pads in these species. To our surprise, we found striking similarities in toe pad development between all of the pad-bearing species we examined. We found that toe pads develop after digit webbing recesses. In all pad-bearing species, ridges that become the adhesive scansors and lamellae first form in the distal half of the digit. Throughout development, new ridges begin forming in the proximal direction while the previous ridges begin to grow laterally. Elaborations and derivations in toe pad form, such as bifurcation, occur in the latter-half of embryonic development. The presumably ancestral pattern of plantar scale development we observed in our leopard gecko and fence lizard embryos (both species lacking adhesive digits) demonstrated that scale ridges form all at once along the length of the digit. These differences are similar to those documented between developing non-padded gecko tails and padded tails of crested geckos. This means that anoles and geckos have converged on a similar developmental process! We suggest that toe pads are initially formed through a major repatterning of digital development and then variation is achieved through relatively minor “tinkering,” through either timing or location of developmental patterns.

Scanning electron micrographs (SEMs) of embryonic lizard digit development, progressing from early development (left) to late development (right). The pad-bearing brown anole (Anolis sagrei) and mourning gecko (Lepidodactylus lugubris) have converged on scansor ridges forming in a distal-to-proximal direction, while the paddles leopard gecko (Eublepharis macularius) has scale rows forming all at once along the length of the digit. Lizard photos courtesy of Dr. Stuart Nielsen.

This is by no means the end of this story. We’ve just scratched the surface and there are a several directions to head in. A logical next step is to characterize histological organization through toe pad development. From there, characterizing the genes involved in toe pad morphogenesis, in tandem with the possibilities of new gene editing technologies, would allow us to test mechanisms of toe pad formation and how variation is generated. And, of course, characterizing toe pad development in other species (such as the secondarily padless Anolis onca) may elucidate further conservation or derivation from the trends we found. This is an exciting time to be a toe pad biologist!

Joe Macedonia

Where do you work and what do you do?  

 I’m a retired Associate Professor at Florida Southern College in Lakeland, FL. While at FSC from 2007 through 2016, I conducted research and taught courses in animal behavior, zoology, evolution, and ecology, as well as a capstone course in undergraduate research. I also took students to Jamaica and Bermuda to conduct field research on anoles.

What aspects of anole biology do you study, and what have you learned? 

 I’ve worked mainly on the production and perception of color and motion displays. Most of this research has been conducted collaboratively with my colleague Dave Clark and has been experimental in nature, e.g., video playbacks and anole robots. Leo Fleishman’s work on Anolis sensory ecology has been a major influence on my thinking about how lizards perceive their color displays. I’ve learned that working on anole behavior can be challenging and that, in fact, most experiments fail! But I’ve also learned that it is well worth the effort in the end.

How and why did you start studying anoles? 

 When I was a kid growing up in the 1950’s and 1960’s in central Pennsylvania, individual Anolis carolinensis used to be sold at the circus in little “animal cracker” boxes. I still remember bringing one home. My father built a small cage for it and tirelessly caught insects for it. Much later, as a postdoc at U.C. Davis in 1992, I was reintroduced to magic of anoles by none other than Jonathan Losos. At that time, Jonathan was a postdoc with Tom Schoener and heard about the video playback work that we were doing in Peter Marler’s Lab. Jonathan was interested in figuring out two things: First, would anoles respond to video recordings of other anoles displaying? And second, if they did respond to video, could anoles discriminate conspecific from heterospecific displays? With our Marler Lab colleague Chris Evans, we showed that A. marcanoi males spent more time displaying in synchrony with video clips of conspecific male displays than heterospecific male (A. cybotes) displays. Soon after that, Judy Stamps and I conducted an even more successful video playback experiment on species recognition, in which we used Anolis grahami from Jamaica as subjects.

What do you love most about studying anoles? 

 What’s not to love? I recall Chris Evans calling them “magnificent beasts”, as well as Duncan Irschick referring to them as “mini gods”. Anoles are endlessly fascinating, and you could never run out of species to research.

What is your favorite anole species? 

 Regarding species that I’ve worked with personally, Anolis grahami is probably my favorite. They are reliable performers in behavioral experiments and are always up to the challenge of responding to another anole (or a video of one, or an anole robot!). There are two runners up, however. Anolis conspersus with its blue dewlap (actually a UV-reflecting dewlap whose wavelengths extend into the blue range) is quite photogenic, as a number of contributors to Anole Annals have noted. The other runner up would be Anolis extremus. Their complex color pattern is exceptional, and although they are frustratingly squirrelly to approach in the field, male tail lifting contests can be spectacular. Anolis extremus really are extreme!

Where can people learn more about you and follow you online? 

Website: www.macedonialab.com

Lindsey Swierk

Where do you work and what do you do?  

I am an Assistant Research Professor in the Department of Biological Sciences at Binghamton University, State University of New York. I am also the Associate Director of Research of the Amazon Conservatory for Tropical Studies, outside Iquitos, Peru. My research group studies ecology at the organismal level, with a focus on behavior and herpetology. I primarily conduct fieldwork in Costa Rica and Peru. At the university, I teach courses in animal behavior and ecology, and I am involved with initiatives to promote underrepresented students in biology. I was a postdoctoral fellow at Yale University and received my PhD in Ecology from Penn State in 2013.

What aspects of anole biology do you study, and what have you learned? 

 My background in behavioral ecology first nudged my interest in anoles towards communication and reproductive behavior. I have a particular interest in understanding how sexual signals function and evolve. My group studies anole dewlaps and their costs and benefits. The use of color and patterns, both on the dewlap and the entire body, is also a focus in my group. We use both observational and experimental techniques to better understand how anoles use sexual coloration and behavioral displays to maximize fitness. We’ve found that there are some significant risks posed by bearing conspicuous sexual signals, and that there is a variety of ways in which body color can be used plastically to benefit anoles.

Our work is based on natural history observations, and so we tend to follow where the anoles lead. Lately, that has led us down the path of examining their antipredator adaptations. We are investigating the fascinating underwater diving and rebreathing behaviors of the semi-aquatic anoles, from ecological and physiological perspectives. We documented that some semi-aquatic anole species spend considerable time underwater when pursued. Our interest in antipredator strategies includes a focus on the mechanics of escaping predators, whether that is by swimming, diving, running, or leaping. Semi-aquatic anoles are also remarkably cold tolerant and have very low body temperatures, and so we also investigate their thermal ecology and possible effects of climate change. 

How and why did you start studying anoles? 

 Allergies, initially! I kept anoles as pets almost my entire childhood because a cat or dog was out of the question. Back then, I spent an embarrassing number of hours just watching what anoles did in their tanks. This cemented anoles in my head as the coolest possible lizards Years later, after spending all of my research life studying other cool herps, I stumbled upon anoles again on a teaching trip to Costa Rica. From the moment I saw Anolis aquaticus in the streams, they completely captured my attention. Their habitat and behaviors were so unique from what I had learned about anoles, I knew I had to make them a priority.

What do you love most about studying anoles? 

Two things. Their remarkable adaptations – how evolution has so astoundingly shaped them to particular environments. I love how much is known, but also how much is still unknown about their morphological, physiological, and behavioral traits – I love the element of surprise! Second, I’m still fascinated by simply watching their interactions with one another, which I could do for hours. 

What is your favorite anole species? 

 I absolutely have a favorite – Anolis aquaticus. They’re such quirky representatives of the mere handful of semi-aquatic anoles out there.

Where can people learn more about you and follow you online? 

Website: www.lindseyswierk.com

Twitter: @LindseySwierk

Instagram: lindseyswierk

SICB 2022: Repeatability and Correlation in Thermal Traits

As we know, anoles are ectotherms which could spell trouble under a changing climate. By closely relying on the temperature of their environment to regulate all sorts of physiological processes, anoles may be at risk when the environmental temperature shifts due to climate change. Because of this, many studies have measured traits which describe a lizard’s thermal physiology, and two popular traits are the lizard’s preferred temperature (Tpref) and heat tolerance. But few studies have looked at a) how repeatable these traits are in any individual and b) whether the two are correlated!

Shannan Yates stands next to her SICB poster.

Shannan with her poster at SICB 2022 in Phoenix.

Shannan Yates, a graduate student at Tulane University in Dr. Alex Gunderson’s lab endeavored to do just this, and presented the results at SICB this year with a very compelling poster. Shannan hypothesized that if Tpref was measured twice in the same individual, that these temperatures should be repeatable. Secondly, she hypothesized that preferred temperature would be correlated with heat tolerance, as these traits are expected to be phenotypically linked, and individuals with high heat tolerances should prefer higher temperatures.

Interestingly, though, Shannan found quite the opposite! Tpref was neither repeatable nor correlated with heat tolerance! This has important implications for many studies which attempt to quantify these traits in Anolis lizards. Shannan concluded that either thermal preference is flexible, or that the current methods used to study Tpref may affect our measurements. Also, thermal preference may not actually be correlated with thermal tolerance at an individual level, or our current methods are indeed obscuring a relationship.

Shannan’s poster is up on SICB+, so be sure to check out the data for yourself, and check out Shannan on twitter here

Aaron Alcala

Where do you work and what do you do?  

 I am a PhD candidate in the Department of Genetics at the University of Georgia. I am doing my thesis research in the lab of Dr. Doug Menke. My research is focused on studying how genes are regulated to build parts of the body during embryonic development, and how changes during embryogenesis can lead to the evolution of novel forms and structures.

What aspects of anole biology do you study, and what have you learned? 

 I am studying the genetic and developmental processes underlying the morphology of limbs in anoles. Although the ecology and evolution of these lizards has been extensively studied, we know relatively little about the underlying mechanisms that have contributed to the evolution of different structures within this genus. One of the most apparent differences among anole species is limb size, which is adapted for the particular microhabitat that each species resides within. Differences observed in adult limb lengths between several species of anoles seem to be due to changes that occur early in embryonic development. I’ve learned that since the genes and signaling pathways underlying limb development are highly conserved, changes in gene regulation may underlie many of the differences in limb morphology observed between species. My current work is focused on comparing DNA sequences in the genomes of different species to find regions involved in regulating genes important for the development and evolution of limb morphology.

How and why did you start studying anoles? 

When I first joined the Menke lab at UGA, our efforts were transitioning into more studies in anoles. I instantly fell in love with the brown anole as a model organism for studies of evolutionary developmental (evo-dev) biology.

What do you love most about studying anoles? 

 For the past few decades, studies of gene function in reptiles have lagged behind other major vertebrate groups. I enjoy being able to contribute to establishing the brown anole as an emerging model organism to investigate the genetic and molecular mechanisms of evolution in reptiles. I am also happy to get out of the lab every once in a while to catch anoles and bring them back for us to study!

What is your favorite anole species? 

 Tough to choose of course, but Anolis grahami is one of my favorites because of its striking colors of greens and blues. 

Where can people learn more about you and follow you online? 

Website: aaronevodevo.wixsite.com/aaronevodevo

Twitter: @aaronevodevo

Instagram: @aaronevodevo

Michele Johnson

Where do you work and what do you do?

 I work at Trinity University in San Antonio, Texas. I teach courses on evolution and vertebrate biology, and I lead a lab of undergraduate scientists studying lizard behavior and physiology. We combine approaches from ecology, evolution, and neuroscience to understand how and why lizards behave the way they do.

What aspects of anole biology do you study, and what have you learned? 

 One of the main areas of my research is the evolution of behavioral mechanisms, and we generally focus on how muscles allow different species to behave in different ways.  A surprising result from this work has been that anole species that use a muscle frequently don’t usually have larger muscles than species that use the muscle rarely. Instead, we found that muscles that move bigger structures are bigger than muscles that move smaller structures, no matter how often the muscle is used. This highlights how different lizard muscles seem to be from muscles in mammals and birds.

Another aspect of our work focuses on how a lizard’s social and physical environment affects how it behaves. We’ve studied why green anoles change their body color, how anoles communicate to form a social hierarchy, how anoles respond to artificial light at night, and many other questions. 

How and why did you start studying anoles? 

 In college, I became fascinated with studying evolution on islands, because I thought islands were where the most exciting evolutionary stories were being discovered. I had never studied anoles before graduate school, but since I joined Jonathan Losos’ lab as a PhD student, they’ve been the focus of almost all my research.

What do you love most about studying anoles? 

 As a behavioral ecologist, my favorite part of studying anoles is watching what they do in the field. I’ve found that observing a single lizard for an hour can often lead to surprising findings, and by combining lots of those observations, we can get a rich understanding of how lizards interact with each other.

What is your favorite anole species? 

 My favorite anole is Anolis bahorucoensis, a lizard that lives in montane forests in the Dominican Republic. They have tiny dewlaps, but their bodies are so vibrantly colored – green and blue and black and yellow and orange.

Where can people learn more about you and follow you online? 

Lab Website: www.johnsonlizardlab.org

Outreach Website: www.lizardsandfriends.org

Twitter: @LizardMichele

Bob Trivers’ Early Years

Bob Trivers published his memoirs, Wild Life, six years ago. We discussed it in these pages and pointed to a favorable book review that appeared in Current Biology, a review with which I agree completely, not surpisingly, since I wrote it.

I commend the book to you, but if you want the short story, check out Trivers’ two-page summary of his life from age 13 to 29 just published in Evolutionary Psychology. The abstract is a good indication of what the short paper holds: “This is a brief history of my intellectual life from age 13 to 29 years—and beyond. It encompasses mathematics, US history, and evolutionary biology, especially social theory based on natural selection.”

Surveying Claw and Toepad Diversity Across Anole Adaptive Radiations

An Anolis biporcatus enjoys some sun while balancing along a branch with the help of those neat adhesive toepads and claws.  Credit

An Anolis biporcatus enjoys some sun while balancing along a branch with the help of those neat adhesive toepads and claws.  Credit Wikimedia Commons.

In a recently published study, Cortés-Parra et al. (2021) compare the claws and toepads of mainland and island Anolis radiations in examination of “morphotype” diversity across phylogenetic and ecological scales. This study intersects nicely with quite recent work comparing the mainland and island radiations (e.g., Hiue et al. 2021; Patton et al. 2021), and claw morphology (e.g., Yuan et al. 2019, 2020; Falvey et al. 2020).

New literature alert!

Claws and toepads in mainland and island Anolis (Squamata: Dactyloidae): Different adaptive radiations with intersectional morphospatial zones

In Journal of Anatomy

Cortés-Parra, Calderón-Espinosa, and Jerez

Abstract:

Anolis lizards have evolved morphologies in response to different selective factors related to microhabitat use. Morphological diversity exhibits evolutionary patterns that reveal similarities and unique regional traits among the mainland and island environments and among Greater Antilles and Lesser Antilles islands. In the Greater Antilles and mainland environments anole species are classified into morphological/ecological groups, that are known as morphotypes (mainland) or ecomorphs (Greater Antilles). Morphotypes are defined only with morphological information; in contrast, for ecomorph assignment both morphology and ethology are required. For mainland species distributed in northwestern South America 10 morphotypes were proposed to include the morphological diversity of 59 species. We obtained data from body size, limbs length, tail length, and the number of lamellae for an additional ten species occurring in the same region and assigned them into morphotypes. We also collected data of the claw and toepad diversity of mainland and island Anolis from northwestern South America and compared it to the claw and toepads morphology recorded for the Greater Antilles and Lesser Antilles islands, under a phylogenetic framework. We found new island morphotypes (MT11–MT13) of Anolis from northwestern South America. When comparing claws and toepads morphology among the 13 morphotypes we found that morphological variation of these traits partially corresponds to morphotype groups. For instance, habitat specialist species like Anolis heterodermus, classified in morphotype 4 (MT4), have a characteristic design of broad toepad and reduced claws, and non-unique design of toepads and claws occurs in morphotypes MT1, MT2, MT5, MT10, and MT13. We also compared claws and toepads of fore and hindlimbs within the same individual, and found that even if limbs show differences in claws and toepads, suggesting that they perform differential biomechanical function, the degree of within individual variation is specific and not related to morphotype assignment. Our data supported the convergent and unique regional evolution among mainland and island anoles, and revealed aspects of correlative evolution of functional traits of claws and toepads that probably are related to minor differences in microhabitat use among mainland and island species, as suggested by previously published literature. Lastly, the evolutionary pattern of morphological diversity of claws and toepads of Anolis in the mainland and island environment supports both unique regional traits and common selective and historical factors that have molded Anolis morphological diversity.

Literature Cited:

Cortés‐Parra, C., Calderón‐Espinosa, M. L., & Jerez, A. (2021). Claws and toepads in mainland and island Anolis (Squamata: Dactyloidae): Different adaptive radiations with intersectional morphospatial zones. Journal of anatomy.

Falvey, C. H., Aviles-Rodriguez, K. J., Hagey, T. J., & Winchell, K. M. (2020). The finer points of urban adaptation: intraspecific variation in lizard claw morphology. Biological Journal of the Linnean Society, 131(2), 304-318.

Huie, J. M., Prates, I., Bell, R. C., & de Queiroz, K. (2021). Convergent patterns of adaptive radiation between island and mainland Anolis lizards. Biological Journal of the Linnean Society, 134(1), 85-110.

Patton, A. H., Harmon, L. J., del Rosario Castañeda, M., Frank, H. K., Donihue, C. M., Herrel, A., & Losos, J. B. (2021). When adaptive radiations collide: Different evolutionary trajectories between and within island and mainland lizard clades. Proceedings of the National Academy of Sciences118(42).

Yuan, M. L., Wake, M. H., & Wang, I. J. (2019). Phenotypic integration between claw and toepad traits promotes microhabitat specialization in the Anolis adaptive radiation. Evolution73(2), 231-244.

Yuan, M. L., Jung, C., Wake, M. H., & Wang, I. J. (2020). Habitat use, interspecific competition and phylogenetic history shape the evolution of claw and toepad morphology in Lesser Antillean anoles. Biological Journal of the Linnean Society129(3), 630-643.

SICB 2022: Anole Nesting Behavior under Predator-Presence!

Dr. Jenna Pruett with the study organism from her SICB talk, Anolis sagrei .

While nesting is ubiquitous across taxa, Dr. Jenna Pruett is interested (like many of us on this site) in non-avian reptiles. Dr. Pruett is currently and NSF Postdoctoral Fellow at University of Colorado – Boulder under Dr. Ambika Kamath and completed her Ph.D. with Dr. Daniel Warner at Auburn University in 2021. For her doctoral work, Dr. Pruett studied nesting behavior in the Cuban brown anole (Anolis sagrei). She found that A. sagrei females tend to nest on the ground, under cover objects (e.g., under rocks), and in areas with high soil moisture and lower temperatures than compared to ambient surroundings. Interestingly, she found that these nest sites were correlated with positive effects on offspring survival.

At SICB 2022, Dr. Pruett discussed a chapter of dissertation research where she was interested in other environmental variables that might influence nesting behavior, such as the presence of predators. The northern curly-tailed lizard (Leiocephalus carinatus) has a shared evolutionary history (i.e., native to Bahamas, invasive in Florida) with A. sagrei. Additionally, A. sagrei is considered a trunk-ground species and L. carinatus is mostly ground-dwelling. Other research suggests that when L. carinatus is present, individual A. sagrei tend to perch higher in the canopy and an increase in mortality is observed (particularly for females).

Dr. Pruett’s experimental set-up from this project.

To address whether predation risk influences A. sagrei nesting behavior, Dr. Pruett designed walk-in cages that were visually separated from each other and contained two nesting options for females: (1) a nest pot above the ground and (2) a nest pot on the ground in a cage with L. carinatus. Some of these cages contained a predator, while some did not. She predicted that females would prefer to nest in above-ground areas when predators were present.

Dr. Pruett found that overall, females tended to prefer laying eggs on the ground prior to predator presence. After predators were added to cages, there was a distinct difference in nesting behavior in cages with and without a predator (i.e., over 50% of eggs laid were above ground when a predator was present). Lastly, when predators were removed, there was a continued upward trend of laying eggs above ground in both treatments. Dr. Pruett suggests that this might result because above-ground pots are a relatively pristine environment to nest, and that if given that option under natural conditions, females might tend to nest above ground as well. Additionally, the above-ground pot may have also gotten more sunlight and more/less moisture that might account for female preference. Dr. Pruett also adds that lizards in cages that were without a predator may have sensed that L. carinatus were in the area (i.e., through mechanisms other than visual) that led them to nest higher.

Dr. Pruett concluded her SICB talk with three remarks: 1) there is an effect of predator presence on A. sagrei nesting behavior, 2) there is an increased preference for above-ground nests in both treatment groups that could be due to other environmental factors, and 3) there need to be more studies on anole nesting, which is a critical component of reproduction, but is understudied in many species. Dr. Pruett is one of the world’s leading researchers in anole nesting behavior and I would highly encourage you to reach out to her with any questions!

“SICB has provided invaluable networking opportunities for me! I met my PhD advisor there, and each year it’s an opportunity to see current research and exchange exciting ideas,” Dr. Pruett says of SICB.

Check out Dr. Pruett’s talk here! Check out Dr. Pruett’s twitter page here!

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