Scott M. Boback

A Developmental Staging Series for the African House Snake, Boaedon (Lamprophis) fuliginosus

Embryonic staging series are important tools in the study of morphological evolution as they establish a common standard for future studies. In this study, we describe the in ovo embryological development of the African house snake (Boaedon fuliginosus), a non-venomous, egg-laying species within the superfamily Elapoidea. We develop our staging series based on external morphology of the embryo including the head, eye, facial prominences, pharyngeal slits, heart, scales, and endolymphatic ducts. An analysis of embryonic growth in length and mass is presented, as well as preliminary data on craniofacial skeletal development. Our results indicate that B. fuliginosus embryos are well into organogenesis but lack well-defined facial prominences at the time of oviposition. Mandibular and maxillary processes extend rostrally within 8 days (stage 3), corresponding to the first appearance of Meckels cartilages. Overall, the development of the craniofacial skeleton in B. fuliginosus appears similar to that of other snake species with intramembraneous bones (e.g., dentary and compound bones) ossifying before most of the endochondral bones, the first of which to ossify are the quadrate and the otic capsule. Our staging series is the first to describe the post-ovipositional development of a non-venomous elapoid based on external morphology. This species is an extremely tractable captive that can produce large clutches of eggs every 45 days throughout the year. As such, B. fuliginosus should be a good model for evolutionary developmental biologists focusing on the craniofacial skeleton, loss of limbs, generational teeth, and venom delivery systems.

Phylogeographic and population genetic analyses reveal multiple species of Boa and independent origins of insular dwarfism

Boa is a Neotropical genus of snakes historically recognized as monotypic despite its expansive distribution. The distinct morphological traits and color patterns exhibited by these snakes, together with the wide diversity of ecosystems they inhabit, collectively suggest that the genus may represent multiple species. Morphological variation within Boa also includes instances of dwarfism observed in multiple offshore island populations. Despite this substantial diversity, the systematics of the genus Boa has received little attention until very recently. In this study we examined the genetic structure and phylogenetic relationships of Boa populations using mitochondrial sequences and genome-wide SNP data obtained from RADseq. We analyzed these data at multiple geographic scales using a combination of phylogenetic inference (including coalescent-based species delimitation) and population genetic analyses. We identified extensive population structure across the range of the genus Boa and multiple lines of evidence for three widely-distributed clades roughly corresponding with the three primary land masses of the Western Hemisphere. We also find both mitochondrial and nuclear support for independent origins and parallel evolution of dwarfism on offshore island clusters in Belize and Cayos Cochinos Menor, Honduras.

Snake modulates constriction in response to prey's heartbeat

Many species of snakes use constriction—the act of applying pressure via loops of their trunk—to subdue and kill their prey. Constriction is costly and snakes must therefore constrict their prey just long enough to ensure death. However, it remains unknown how snakes determine when their prey is dead. Here, we demonstrate that boas (Boa constrictor) have the remarkable ability to detect a heartbeat in their prey and, based on this signal, modify the pressure and duration of constriction accordingly. We monitored pressure generated by snakes as they struck and constricted warm cadaveric rats instrumented with a simulated heart. Snakes responded to the beating heart by constricting longer and with greater total pressure than when constricting rats with no heartbeat. When the heart was stopped midway through the constriction, snakes abandoned constriction shortly after the heartbeat ceased. Furthermore, snakes naive to live prey also responded to the simulated heart, suggesting that this behaviour is at least partly innate. These results are an example of how snakes integrate physiological cues from their prey to modulate a complex and ancient behavioural pattern.

Variation in Color and Color Change in Island and Mainland Boas (Boa constrictor)

Abstract Physiological color change and geographic variation in coloration are well documented in several squamate lineages, having presumably evolved for cryptic, sexual signaling, and thermoregulatory functions. Only 11 snake species have demonstrated physiological color change, although anecdotal reports suggest it may be present in additional species. We describe color variation and physiological color change in island and mainland populations of Boa constrictor using full-spectrum reflectance spectrometry. We employed principal components analysis (PCA) summarizing the spectrometry data into two axes that describe the brightness (PC1) and chroma or relative intensity of particular wavelengths (PC2). Boas from island and mainland localities exhibited physiological color change, and this change occurred on a diel cycle. Boas from both locations were lighter at night and darker during the day. The magnitude of the color change differed between our two PC axes. Although change in brightness was similar for boas on the islands and the mainland, the change in chroma was greater in boas from the mainland. Color also varied seasonally; boas were lighter in color and reflected more long-wavelength light during the wet season than during the dry season in Belize. We suggest that a fundamental hormone cycle (melatonin/melanophore stimulating hormone, MSH) present in a wide variety of vertebrates, underlies the physiological color change in snakes. If this is true, color change may be more widespread than previously realized, and the perceptual bias of the human vision system may have caused researchers to discount its presence in snakes.

Snake constriction rapidly induces circulatory arrest in rats

ABSTRACT As legless predators, snakes are unique in their ability to immobilize and kill their prey through the process of constriction, and yet how this pressure incapacitates and ultimately kills the prey remains unknown. In this study, we examined the cardiovascular function of anesthetized rats before, during and after being constricted by boas (Boa constrictor) to examine the effect of constriction on the preys circulatory function. The results demonstrate that within 6 s of being constricted, peripheral arterial blood pressure (PBP) at the femoral artery dropped to 1/2 of baseline values while central venous pressure (CVP) increased 6-fold from baseline during the same time. Electrocardiographic recordings from the anesthetized rats heart revealed profound bradycardia as heart rate (fH) dropped to nearly half of baseline within 60 s of being constricted, and QRS duration nearly doubled over the same time period. By the end of constriction (mean 6.5±1 min), rat PBP dropped 2.9-fold, fH dropped 3.9-fold, systemic perfusion pressure (SPP=PBP−CVP) dropped 5.7-fold, and 91% of rats (10 of 11) had evidence of cardiac electrical dysfunction. Blood drawn immediately after constriction revealed that, relative to baseline, rats were hyperkalemic (serum potassium levels nearly doubled) and acidotic (blood pH dropped from 7.4 to 7.0). These results are the first to document the physiological response of prey to constriction and support the hypothesis that snake constriction induces rapid prey death due to circulatory arrest. Highlighted Article: Examination of the effect of snake constriction on rat cardiovascular function reveals support for circulatory arrest as the proximate cause of death in constricted prey, in contrast to previous ideas.

Divergence in morphology, but not habitat use, despite low genetic differentiation among insular populations of the lizard Anolis lemurinus in Honduras

Studies of recently isolated populations are useful because observed differences can often be attributed to current environmental variation. Two populations of the lizard Anolis lemurinus have been isolated on the islands of Cayo Menor and Cayo Mayor in the Cayos Cochinos Archipelago of Honduras for less than 15000 y. We measured 12 morphometric and 10 habitat-use variables on 220 lizards across these islands in 2 y, 2008 and 2009. The goals of our study were (1) to explore patterns of sexual dimorphism, and (2) to test the hypothesis that differences in environment among islands may have driven divergence in morphology and habitat use despite genetic homogeneity among populations. Although we found no differences among sexes in habitat use, males had narrower pelvic girdles and longer toe pads on both islands. Between islands, males differed in morphology, but neither males nor females differed in habitat use. Our data suggest that either recent selection has operated differentially on males despite low genetic differentiation, or that they display phenotypic plasticity in response to environmental variation. We suggest that patterns may be driven by variation in intrapopulation density or differences in predator diversity among islands.

DISTRIBUTION, HABITAT, SIZE, AND COLOR PATTERN OF CNEMIDOPHORUS LEMNISCATUS (SAURIA: TEIIDAE) ON CAYO COCHINO PEQUENO, HONDURAS

Abstract Cayo Cochino Pequeño is a 0.64-km2 Caribbean island in the Cayos Cochinos archipelago, Department of Islas de la Bahía, Honduras. One published report noted the presence of the rainbow whiptail (Cnemidophorus lemniscatus) on Cayo Cochino Pequeño, but nothing is known about the biology of this insular population. During a part of the dry season in July and August 2004, we used drift fences, pitfall traps, and separate observational transects to elucidate the distribution and habitat use of C. lemniscatus on the island. The only population of this species was located in a narrow coastal zone (width to 60 m and length to 450 m) on the southern half of the eastern windward side of the island. We analyzed the percentage of the canopy cover and the percentage of 4 ground coverage types along 2 transects 200 m long in this area to understand the basis of the suitability of the habitat for C. lemniscatus. Descriptively, the area harboring this species on Cayo Cochino Pequeño consisted of the remnants of a coconut palm grove with low-lying herbaceous vegetation and grasses, in which a mosaic of small, open areas of sandy soil and coral fragments, with or without accumulations of debris, were the foci of lizard activities. Also observed in this habitat were 2 individuals of the brown racer (Dryadophis melanolomus), an actively foraging snake and likely predator on C. lemniscatus. Data obtained on rainbow whiptails captured in pitfall traps and subsequently released were used to determine the size and color patterns of hatchlings and adult males and females.

Carotenoid Composition of Colorful Body Stripes and Patches in the Painted Turtle (Chrysemys picta) and Red-Eared Slider (Trachemys scripta)

Abstract The objective quantification of integumentary color and its pigmentary basis have been relatively little studied in turtles. We used reflectance spectrometry to measure the color of the chin, neck, and leg stripes and spots of the painted turtle (Chrysemys picta) and the red-eared slider (Trachemys scripta), and we used absorbance spectrometry and high-performance liquid chromatography to identify pigments in these colored patches. Both species had stripes and patches that were predominantly yellow and orange, but the yellow stripes typically contained a small ultraviolet (UV) reflectance peak. Chin, neck, and leg stripes of the painted turtle and chin and neck stripes of the red-eared slider contained an unidentified apocarotenoid and a ketocarotenoid (astaxanthin), but the postorbital eyespot of the red-eared slider contained only astaxanthin. These findings add painted turtles and red-eared sliders to the growing list of animals that use carotenoids in part to generate red, orange, yellow, and UV colors. We discuss these findings in the context of dual pigment production, pigment metabolic production costs, immune system health, and visual signal use in emydid turtles and other vertebrates.

Differential Disease Susceptibilities in Experimentally Reptarenavirus-Infected Boa Constrictors and Ball Pythons

ABSTRACT Inclusion body disease (IBD) is an infectious disease originally described in captive snakes. It has traditionally been diagnosed by the presence of large eosinophilic cytoplasmic inclusions and is associated with neurological, gastrointestinal, and lymphoproliferative disorders. Previously, we identified and established a culture system for a novel lineage of arenaviruses isolated from boa constrictors diagnosed with IBD. Although ample circumstantial evidence suggested that these viruses, now known as reptarenaviruses, cause IBD, there has been no formal demonstration of disease causality since their discovery. We therefore conducted a long-term challenge experiment to test the hypothesis that reptarenaviruses cause IBD. We infected boa constrictors and ball pythons by cardiac injection of purified virus. We monitored the progression of viral growth in tissues, blood, and environmental samples. Infection produced dramatically different disease outcomes in snakes of the two species. Ball pythons infected with Golden Gate virus (GoGV) and with another reptarenavirus displayed severe neurological signs within 2 months, and viral replication was detected only in central nervous system tissues. In contrast, GoGV-infected boa constrictors remained free of clinical signs for 2 years, despite high viral loads and the accumulation of large intracellular inclusions in multiple tissues, including the brain. Inflammation was associated with infection in ball pythons but not in boa constrictors. Thus, reptarenavirus infection produces inclusions and inclusion body disease, although inclusions per se are neither necessarily associated with nor required for disease. Although the natural distribution of reptarenaviruses has yet to be described, the different outcomes of infection may reflect differences in geographical origin. IMPORTANCE New DNA sequencing technologies have made it easier than ever to identify the sequences of microorganisms in diseased tissues, i.e., to identify organisms that appear to cause disease, but to be certain that a candidate pathogen actually causes disease, it is necessary to provide additional evidence of causality. We have done this to demonstrate that reptarenaviruses cause inclusion body disease (IBD), a serious transmissible disease of snakes. We infected boa constrictors and ball pythons with purified reptarenavirus. Ball pythons fell ill within 2 months of infection and displayed signs of neurological disease typical of IBD. In contrast, boa constrictors remained healthy over 2 years, despite high levels of virus throughout their bodies. This difference matches previous reports that pythons are more susceptible to IBD than boas and could reflect the possibility that boas are natural hosts of these viruses in the wild.

Toxic, invasive treefrog creates evolutionary trap for native gartersnakes

Possession of unique defensive toxins by nonindigenous species may increase the likelihood of creating evolutionary traps for native predators. We tested the hypothesis that nonindigenous, toxic Cuban Treefrogs (Osteopilus septentrionalis) have created an evolutionary trap for native, generalist snakes. Additionally, we explored the possibility that populations of snakes that co-occur with Cuban Treefrogs have responded in ways that allow them to escape potential trap dynamics. To evaluate a potential fitness cost of consuming Cuban Treefrogs, we monitored growth of 61 wild-caught Common Gartersnakes (Thamnophis sirtalis) fed exclusive diets of either Cuban Treefrogs, native Green Treefrogs (Hyla cinerea), or native Golden Shiners (Notemigonus crysoleucas). Snakes in the Cuban Treefrog diet treatment gained less than half the mass of those consuming native prey, and Cuban Treefrogs were significantly less digestible than native prey. There was no difference in the response of gartersnakes to prey scent cues of Cuban Treefrogs and Green Treefrogs. Our results indicate that Cuban Treefrogs likely represent an evolutionary trap for snakes because consumption of these frogs carries fitness costs, yet snakes fail to recognize this prey as being costly. We found no difference in growth or response to prey cues between snakes from invaded and non-invaded regions, suggesting snakes have not responded to escape trap dynamics. Interactions of native snakes and Cuban Treefrogs support the idea that introduced species with novel toxins may increase the likelihood of evolutionary trap formation.