Drift,selection and adaptive variation in small populations of a threatened rattlesnake

An important goal of conservation genetics is to determine if the viability of small populations is reduced by a loss of adaptive variation due to genetic drift. Here, we assessed the impact of drift and selection on direct measures of adaptive variation (toxin loci encoding venom proteins) in the eastern massasauga rattlesnake (Sistrurus catenatus), a threatened reptile that exists in small isolated populations. We estimated levels of individual polymorphism in 46 toxin loci and 1,467 control loci across 12 populations of this species, and compared the results with patterns of selection on the same loci following speciation of S. catenatus and its closest relative, the western massasauga (S. tergeminus). Multiple lines of evidence suggest that both drift and selection have had observable impacts on standing adaptive variation. In support of drift effects, we found little evidence for selection on toxin variation within populations and a significant positive relationship between current levels of adaptive variation and long‐ and short‐term estimates of effective population size. However, we also observed levels of directional selection on toxin loci among populations that are broadly similar to patterns predicted from interspecific selection analyses that pre‐date the effects of recent drift, and that functional variation in these loci persists despite small short‐term effective sizes. This suggests that much of the adaptive variation present in populations may represent an example of “drift debt,” a nonequilibrium state where present‐day levels of variation overestimate the amount of functional genetic diversity present in future populations.     

Evaluating Basking-Habitat Deficiency in the Threatened Eastern Massasauga Rattlesnake

ABSTRACT Woody plant succession is hypothesized to threaten many reptile populations by reducing the amount of solar energy available for thermoregulation. Mitigation via vegetation management is often recommended; however, the need for such management practices rarely has been evaluated. We examined the need for basking-site enhancement for the eastern massasauga rattlesnake (Sistrurus c. catenatus; hereafter EMR) in New York, USA, where only 2 populations remain: one at an open-canopy site and another at a closed-canopy site. Microhabitat temperatures were substantially lower at the closed-canopy site, where EMRs selected the warmest available basking sites. Eastern massasauga rattlesnakes in the open-canopy population selected basking sites that afforded greater cryptic cover. We recommend experimental reduction of shrub cover to improve EMR basking habitat at the closed-canopy site. More generally, we caution that management efforts to reduce shrub cover for basking EMRs should maintain adequate cryptic cover.     

Range-wide analysis of eastern massasauga survivorship

Decisions affecting wildlife management and conservation policy of imperiled species are often aided by population models. Reliable population models require accurate estimates of vital rates and an understanding of how vital rates vary geographically. The eastern massasauga (Sistrurus catenatus catenatus) is a rattlesnake species found in the Great Lakes region of North America. Populations of the eastern massasauga are fragmented and only a few areas harbor multiple, sizable populations. Eastern massasauga research has typically focused on single populations or local metapopulations but results suggest that demographic parameters vary geographically. We used 21 radiotelemetry datasets comprising 499 telemetered snakes from 16 distinct locations throughout the range of the eastern massasauga to characterize geographic patterns of adult survival using the known-fate model in Program MARK. Annual adult survival ranged from 0.35 to 0.95 (mean = 0.67) and increased along a southwest to northeast geographic axis. Further analysis of 6 datasets indicated no consistent difference in survival between males and females. Our results provide a better understanding of the relationship between survivorship and geography for the eastern massasauga and suggest that such variation should be incorporated into population models as well as local and regional management plans. © 2012 The Wildlife Society.     

Behavioral and life history responses of eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) to human disturbance

Parks and nature reserves protect important natural habitats but also provide public opportunities for outdoor recreational activities that may have unintended negative effects on wildlife. We examined the response of eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) to inadvertent disturbance by humans in Killbear Provincial Park, Ontario, Canada. Radio telemetry of 25 adult snakes over two active seasons revealed that, as disturbance increased, gravid females were less visible to observers, but the visibility of non-gravid females and males did not change. Mean distance moved per day decreased and mean time between moves greater than 10 m increased in gravid females, non-gravid females and males with increasing exposure to human disturbance. However, mark-recapture data revealed no differences in the condition or growth rates of snakes, or in the litter size of gravid females, between individuals captured in disturbed and undisturbed study areas. While it is possible that the behavioral responses we observed are not sufficient to have life history consequences, more detailed information on the exposure of individual snakes to human activity is necessary before the conclusion that disturbance is not detrimental to snakes can be accepted. Similarly, other potential negative effects of human disturbance not investigated here remain to be explored.     

Massasauga Home Range Patterns in the Midwest

Abstract: The massasauga (Sistrurus catenatus) has drastically declined throughout its range mainly due to habitat loss and human persecution. Populations of the massasauga that occur north and east of the Missouri River, USA, are currently candidates for listing under the United States Endangered Species Act. In areas where land managers wish to improve habitat for existing populations or create habitat for repatriation efforts, an understanding of massasauga spatial ecology is necessary to ensure that adequate sized blocks of habitat are created and properly managed. We studied spatial ecology of massasaugas at 2 sites in Wisconsin and 3 sites in Missouri over an 11-year period. Our results indicated that male massasaugas had larger spatial requirements than all other cohorts. Our study sites supporting viable populations indicated that managers interested in restoring or enhancing massasauga habitat should use 100 ha as a minimum restoration or management target. (JOURNAL OF WILDLIFE MANAGEMENT 72(3):754–759; 2008)     

Deconstructing a Complex Molecular Phenotype: Population-Level Variation in Individual Venom Proteins in Eastern Massasauga Rattlesnakes (<Emphasis Type="Italic">Sistrurus c. catenatus</Emphasis>)

Identifying the molecular basis for complex adaptations such as the toxic proteins used by venomous snakes to subdue and digest prey is an important step in understanding the evolutionary and functional basis for such traits. Recent proteomics-based analyses have made possible the identification of all constituent proteins in whole venom samples. Here we exploit this advance to study patterns of population-level variation in venom proteins from 254 adult eastern massasauga rattlesnakes (Sistrurus c. catenatus) collected from 10 populations. Analysis of presence–absence variation in specific proteins from 1D PAGE gels shows that: (1) The frequency spectra for individual protein bands is U-shaped with a large number of specific proteins either being consistently “common” or “rare” across populations possibly reflecting functional differences. (2) Multivariate axes which summarize whole venom variation consist of bands from all major types of proteins implying the integration of functionally distinct components within the overall venom phenotype. (3) There is significant differentiation in venom proteins across populations and the specific classes of proteins contributing to this differentiation have been identified. (4) Levels of population differentiation in venom proteins are not correlated with levels of neutral genetic differentiation, or genetically effective population sizes which argues that patterns of venom variation are not simply a consequence of population structure but leaves open the role of selection in generating population differences in venom. Our results identify particular classes of venom proteins and their associated genes as being fruitful targets for future studies of the molecular and functional basis for this complex adaptive phenotype.     

Biogeographic consequences of shifting climate for the western massasauga (Sistrurus tergeminus)

The western massasauga (Sistrurus tergeminus) is a small pit viper with an extensive geographic range, yet observations of this species are relatively rare. They persist in patchy and isolated populations, threatened by habitat destruction and fragmentation, mortality from vehicle collisions, and deliberate extermination. Changing climates may pose an additional stressor on the survival of isolated populations. Here, we evaluate historic, modern, and future geographic projections of suitable climate for S. tergeminus to outline shifts in their potential geographic distribution and inform current and future management. We used maximum entropy modeling to build multiple models of the potential geographic distribution of S. tergeminus. We evaluated the influence of five key decisions made during the modeling process on the resulting geographic projections of the potential distribution, allowing us to identify areas of model robustness and uncertainty. We evaluated models with the area under the receiver operating curve and true skill statistic. We retained 16 models to project both in the past and future multiple general circulation models. At the last glacial maximum, the potential geographic distribution associated with S. tergeminus occurrences had a stronghold in the southern part of its current range and extended further south into Mexico, but by the mid‐Holocene, its modeled potential distribution was similar to its present‐day potential distribution. Under future model projections, the potential distribution of S. tergeminus moves north, with the strongest northward trends predicted under a climate scenario increase of 8.5 W/m². Some southern populations of S. tergeminus have likely already been extirpated and will continue to be threatened by shifting availability of suitable climate, as they are already under threat from desertification of grasslands. Land use and habitat loss at the northern edge of the species range are likely to make it challenging for this species to track suitable climates northward over time.     

Inferring species-level phylogenies and taxonomic distinctiveness using multilocus data in Sistrurus rattlesnakes

Phylogenetic relationships and taxonomic distinctiveness of closely related species and subspecies are most accurately inferred from data derived from multiple independent loci. Here, we apply several approaches for understanding species-level relationships using data from 18 nuclear DNA loci and 1 mitochondrial DNA locus within currently described species and subspecies of Sistrurus rattlesnakes. Collectively, these methods provide evidence that a currently described species, the massasauga rattlesnake (Sistrurus catenatus), consists of two well-supported clades, one composed of the two western subspecies (S. c. tergeminus and S. c. edwardsii) and the other the eastern subspecies (S. c. catenatus). Within pigmy rattlesnakes (S. miliarius), however, there is not strong support across methods for any particular grouping at the subspecific level. Monophyly based tests for taxonomic distinctiveness show evidence for distinctiveness of all subspecies but this support is strongest by far for the S. c. catenatus clade. Because support for the distinctiveness of S. c. catenatus is both strong and consistent across methods, and due to its morphological distinctiveness and allopatric distribution, we suggest that this subspecies be elevated to full species status, which has significant conservation implications. Finally, most divergence time estimates based upon a fossil-calibrated species tree are > 50% younger than those from a concatenated gene tree analysis and suggest that an active period of speciation within Sistrurus occurred within the late Pliocene/Pleistocene eras.     

Resource selection by the eastern massasauga rattlesnake on managed land in southwestern Michigan

The eastern massasauga rattlesnake (Sistrurus catenatus catenatus) has experienced population declines throughout its range and is now a candidate for federal protection. However, little is known about massasauga habitat selection in Michigan, particularly in actively managed landscapes. Our objectives were to: 1) quantify whether massasaugas in southwestern Michigan select certain vegetation types disproportionately to their availability and 2) quantify whether the vegetation structure associated with snake locations differed between managed (e.g., burning, woody species removal) and unmanaged stands. We implanted radio transmitters in 51 snakes from 2004 to 2005 and 2008 to 2009. We quantified second-order resource selection using compositional analysis, and modeled the effect of habitat management efforts on vegetation using 4 structural variables. All snakes selected cover types disproportionately to their availability (P = 0.001); a ranking matrix ordered the vegetation types, from most to least used, as: early-mid successional deciduous wetland > early-mid successional deciduous upland > developed > late successional mixed lowland forest > late successional deciduous upland forest. We found that snakes in managed areas were associated with greater amounts of dead herbaceous cover (P = 0.005) and less woody stem density (P < 0.001) and tree dominance (P < 0.001) than were snakes in unmanaged areas; however, live herbaceous cover was comparable. Our results can be used by regional managers to provide early and mid successional habitat with structure similar to that selected by snakes in Michigan. © 2011 The Wildlife Society.     

To forage,mate, or thermoregulate: Influence of resource manipulation on male rattlesnake behavior

Male animals should preferentially allocate their time to performing activities that promote enhancing reproductive opportunity, but the need to acquire resources for growth and survival may compete with those behaviors in the short term. Thus, behaviors which require differing movement patterns such as ambushing prey and actively searching for mates can be mutually exclusive. Consequently, males that succeed at foraging could invest greater time and energy into mate searching. We radio‐tracked sixteen male massasauga rattlesnakes (Sistrurus catenatus) and supplemented the diets of half the snakes with mice across an active season. We tested the predictions that reduced foraging needs would allow fed snakes to move (i.e., mate search) more, but that they would consequently be stationary to thermoregulate less, than unfed controls. Contrary to our first prediction, we found no evidence that fed snakes altered their mate searching behavior compared to controls. However, we found controls maintained higher body temperatures than fed snakes during the breeding season, perhaps because fed snakes spent less time in exposed ambush sites. Fed snakes had higher body condition scores than controls when the breeding season ended. Our results suggest the potential costs incurred by devoting time to stationary foraging may be outweighed by the drive to increase mating opportunities. Such instances may be especially valuable for massasaugas and other temperate reptiles that can remain inactive for upwards of half their lives or longer in some cases, and for female rattlesnakes that generally exhibit biennial or more protracted reproductive cycles.     

Common Cutaneous Bacteria Isolated from Snakes Inhibit Growth of <Emphasis Type="Italic">Ophidiomyces ophiodiicola</Emphasis>

There is increasing concern regarding potential impacts of snake fungal disease (SFD), caused by Ophidiomyces ophiodiicola (Oo), on free-ranging snake populations in the eastern USA. The snake cutaneous microbiome likely serves as the first line of defense against Oo and other pathogens; however, little is known about microbial associations in snakes. The objective of this study was to better define the composition and immune function of the snake cutaneous microbiome. Eight timber rattlesnakes (Crotalus horridus) and four black racers (Coluber constrictor) were captured in Arkansas and Tennessee, with some snakes exhibiting signs of SFD. Oo was detected through real-time qPCR in five snakes. Additional histopathological techniques confirmed a diagnosis of SFD in one racer, the species’ first confirmed case of SFD in Tennessee. Fifty-eight bacterial and five fungal strains were isolated from skin swabs and identified with Sanger sequencing. Non-metric multidimensional scaling and PERMANOVA analyses indicated that the culturable microbiome does not differ between snake species. Fifteen bacterial strains isolated from rattlesnakes and a single strain isolated from a racer inhibited growth of Oo in vitro. Results shed light on the culturable cutaneous microbiome of snakes and probiotic members that may play a role in fighting an emergent disease.     

Pathologic and hematologic responses to surgically implanted transmitters in eastern massasauga rattlesnakes (Sistrurus catenatus catenatus)

The study of secretive snakes, such as rattlesnakes, has benefited from the use of radiotelemetry. However, the principal assumption in telemetry studies is that the transmitter has no significant effect on the study animal. To test the validity of this assumption, the physiologic and pathologic effects of intracoelomic implants were examined in a group of 24 eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) in a laboratory setting over a period of 58 wk between March 2005 and April 2006. Inflammation and infection were evaluated using gross examination, histopathology, bacteriology, hematology, and plasma protein electrophoresis. Inflammation and infection occurred despite careful surgical procedures and advanced veterinary care. Four of 12 (33%) snakes developed extensive inflammatory response to the transmitter and associated anaerobic and gram-negative bacterial infections. Another four (33%) snakes showed mild inflammatory responses without infection. Reaction to the transmitters was reflected in changes in values for heterophils, monocytes, alpha-1, and beta globulin levels. Some conclusions reached in field studies using implanted radiotransmitters in snakes may be invalid if the implant influences the behavior or survival of the subject. Advances in attachment methods and transmitter coating technology may prevent some of the adverse effects associated with surgically implanted transmitters.     

Body Size Evolution in Insular Speckled Rattlesnakes (Viperidae: Crotalus mitchellii)

Background

Speckled rattlesnakes (Crotalus mitchellii) inhabit multiple islands off the coast of Baja California, Mexico. Two of the 14 known insular populations have been recognized as subspecies based primarily on body size divergence from putative mainland ancestral populations; however, a survey of body size variation from other islands occupied by these snakes has not been previously reported. We examined body size variation between island and mainland speckled rattlesnakes, and the relationship between body size and various island physical variables among 12 island populations. We also examined relative head size among giant, dwarfed, and mainland speckled rattlesnakes to determine whether allometric differences conformed to predictions of gape size (and indirectly body size) evolving in response to shifts in prey size.

Methodology/Principal Findings

Insular speckled rattlesnakes show considerable variation in body size when compared to mainland source subspecies. In addition to previously known instances of gigantism on Ángel de la Guarda and dwarfism on El Muerto, various degrees of body size decrease have occurred frequently in this taxon, with dwarfed rattlesnakes occurring mostly on small, recently isolated, land-bridge islands. Regression models using the Akaike information criterion (AIC) showed that mean SVL of insular populations was most strongly correlated with island area, suggesting the influence of selection for different body size optima for islands of different size. Allometric differences in head size of giant and dwarf rattlesnakes revealed patterns consistent with shifts to larger and smaller prey, respectively.

Conclusions/Significance

Our data provide the first example of a clear relationship between body size and island area in a squamate reptile species; among vertebrates this pattern has been previously documented in few insular mammals. This finding suggests that selection for body size is influenced by changes in community dynamics that are related to graded differences in area over what are otherwise similar bioclimatic conditions. We hypothesize that in this system shifts to larger prey, episodic saturation and depression of primary prey density, and predator release may have led to insular gigantism, and that shifts to smaller prey and increased reproductive efficiency in the presence of intense intraspecific competition may have led to insular dwarfism.     

The influence of landscape on gene flow in the eastern massasauga rattlesnake (Sistrurus c. catenatus): insight from computer simulations

Understanding how gene flow shapes contemporary population structure requires the explicit consideration of landscape composition and configuration. New landscape genetic approaches allow us to link such heterogeneity to gene flow within and among populations. However, the attribution of cause is difficult when landscape features are spatially correlated, or when genetic patterns reflect past events. We use spatial Bayesian clustering and landscape resistance analysis to identify the landscape features that influence gene flow across two regional populations of the eastern massasauga rattlesnake, Sistrurus c. catenatus. Based on spatially explicit simulations, we inferred how habitat distribution modulates gene flow and attempted to disentangle the effects of spatially confounded landscape features. We found genetic clustering across one regional landscape but not the other, and also local differences in the effect of landscape on gene flow. Beyond the effects of isolation‐by‐distance, water bodies appear to underlie genetic differentiation among individuals in one regional population. Significant effects of roads were additionally detected locally, but these effects are possibly confounded with the signal of water bodies. In contrast, we found no signal of isolation‐by‐distance or landscape effects on genetic structure in the other regional population. Our simulations imply that these local differences have arisen as a result of differences in population density or tendencies for juvenile rather than adult dispersal. Importantly, our simulations also demonstrate that the ability to detect the consequences of contemporary anthropogenic landscape features (e.g. roads) on gene flow may be compromised when long‐standing natural features (e.g. water bodies) co‐exist on the landscape.     

Response of timber rattlesnakes to commercial logging operations

Forest management practices in the eastern United States directly impact large parcels of land that serve as habitat for timber rattlesnakes (Crotalus horridus). We assessed the behavioral response of timber rattlesnakes to commercial logging activities and the impact of such activities on a timber rattlesnake population in northcentral Pennsylvania. We radiotelemetrically monitored 67 individual snakes over periods of up to 4 years, marked and recaptured 306 snakes, and conducted search and survey efforts before, during, and after commercial logging operations on 3 timber sale parcels (totaling 154.2 ha). Location and timing of timber sales created the maximum opportunity for interaction of snakes with logging operations and with altered habitat. Observed logging-related mortality of snakes was low (<2% of the population/yr), but potential mortality could have reached 7%. Logging activity and resulting habitat changes did not alter behavior or movement patterns of telemetrically monitored snakes. Snakes with established activity ranges in timber sale areas continued to use these areas both during and after logging operations. Similarly, snakes with activity patterns that did not include timber sale areas did not alter their movement patterns to include such sites in the short-term. Timbering increased structural diversity of the habitat and, concurrently, diversity of habitat used by timber rattlesnakes increased. Our results suggest that the opportunity exists to develop forest management practices that provide timber products while limiting impacts on behavior and habitat use of timber rattlesnakes. To further reduce impacts to timber rattlesnake populations we recommend that management agencies require commercial logging contractors, sub-contractors, and field employees to adhere strictly to a policy that prohibits the intentional killing of rattlesnakes encountered during logging activities. © 2010 The Wildlife Society.     

Comparative thermal ecology parameters of the mexican dusky rattlesnake (Crotalus triseriatus)

Montane habitats exhibit a high degree of thermal heterogeneity, and thus provide considerable thermoregulatory challenges for ectotherms. Comparative analyses provide an opportunity to understand how variation in abiotic factors (e.g., operative temperatures, thermal quality) can affect life history traits within species. We studied the thermal ecology of three populations of the rattlesnake Crotalus triseriatus inhabiting different volcanoes in the central region of Mexico using the Hertz et al. (1993) protocol. The average body temperature of dusky rattlesnakes from the three study sites was 22.4 °C; mean active body temperature was higher in site 2 than in sites 1 and 3, but no differences between females, males and juveniles nor an interaction among site and sex was found. The thermal quality was low in the three sites, particularly in sites 1 and 3. Thermoregulation accuracy statistically differed among populations: individuals from site 2 were more accurate thermoregulating, while individuals from site 1 were the least accurate. Compared to other snakes, dusky rattlesnakes can be considered as a eurythermic species, which can often be active at relatively low body temperatures.     

Genetic structure of populations of the threatened eastern massasauga rattlesnake, Sistrurus c. catenatus: evidence from microsatellite DNA markers

Throughout its distribution in North America, the threatened eastern massasauga rattlesnake ( Sistrurus c. catenatus ) persists in a series of habitat-isolated disjunct populations of varying size. Here, we use six microsatellite DNA loci to generate information on the degree of genetic differentiation between, and the levels of inbreeding within populations to understand how evolutionary processes operate in these populations and aid the development of conservation plans for this species. Samples were collected from 199 individuals from five populations in Ontario, New York and Ohio. Our results show that all sampled populations: (i) differ significantly in allele frequencies even though some populations are < 50 km apart, and may contain genetically distinct subpopulations < 2 km apart; (ii) have an average of 23% of alleles that are population specific; and (iii) have significant F _IS values (mean overall F _IS= 0.194) probably due to a combination of Wahlund effects resulting from fine-scale genetic differentiation within populations and the presence of null alleles. Our results imply that massasauga populations may be genetically structured on an extremely fine scale even within continuous populations, possibly due to limited dispersal. Additional information is needed to determine if dispersal and mating behaviour within populations can account for this structure and whether the observed differentiation is due to random processes such as drift or to local adaptation. From a conservation perspective, our results imply that these massasauga populations should be managed as demographically independent units and that each has high conservation value in terms of containing unique genetic variation.     

Probing,Assessment, and Management during Interactions between Ground Squirrels and Rattlesnakes

The predator-prey relationship between California ground squirrels (Spermophilus beecheyi) and northern Pacific rattlesnakes (Crotalus viridis) is a useful system for exploring conflict and assessment. Rattlesnakes are major predators of ground squirrel pups, but pose a less significant threat to adult squirrels. Adults approach, harass, and even attack rattlesnakes in defense of their pups. Two factors that may influence risk to both squirrel and snake during encounters are the size and body temperature of the rattlesnake. We used high-speed video to analyze the strikes of rattlesnakes of various sizes tested at different body temperatures. Results indicate that warmer snakes are more dangerous because they strike with higher velocity, greater accuracy, and less hesitation. Similarly, larger snakes are more dangerous because they can strike farther and at higher speeds, and keep their fangs embedded longer. Thus, ground squirrels would benefit from extracting information about a rattlesnake's size and temperature. The converse of our results is that cooler, smaller rattlesnakes may be more vulnerable. These snakes could mitigate their risk by avoiding dangerous adversaries and minimizing cues that divulge their weaknesses. Such tactics might explain the active probing that squirrels direct at rattlesnakes, which may function to overcome a snake's resistance to disclosing its vulnerability.     

Similarity of contemporary and historical gene flow among highly fragmented populations of an endangered rattlesnake

Populations of endangered taxa in recently fragmented habitats often show high levels of genetic structure, but the role that contemporary versus historical processes play in generating this pattern is unclear. The eastern massasauga rattlesnake (Sistrurus c. catenatus) is an endangered snake that presently occurs throughout central and eastern North America in a series of populations that are isolated because of habitat fragmentation and destruction. Here, we use data from 19 species-specific microsatellite DNA loci to assess the levels of genetic differentiation, genetic effective population size, and contemporary and historical levels of gene flow for 19 populations sampled across the range of this snake. Eastern massasaugas display high levels of genetic differentiation (overall θ(Fst) = 0.21) and a Bayesian clustering method indicates that each population represents a unique genetic cluster even at regional spatial scales. There is a twofold variation in genetically effective population sizes but little genetic evidence that populations have undergone recent or historical declines in size. Finally, both contemporary and historical migration rates among populations were low and similar in magnitude even for populations located <7 km apart. A test of alternate models of population history strongly favours a model of long-term drift-migration equilibrium over a recent isolation drift-only model. These results suggest that recent habitat fragmentation has had little effect on the genetic characteristics of these snakes, but rather that this species has historically existed in small isolated populations that may be resistant to the long-term negative effects of inbreeding.     

Snake Population Venomics: Proteomics-Based Analyses of Individual Variation Reveals Significant Gene Regulation Effects on Venom Protein Expression in <Emphasis Type="Italic">Sistrurus</Emphasis> Rattlesnakes

Studies of the molecular basis of adaptations seek to understand the relative importance of structural changes in proteins versus gene regulation effects as determinants of phenotype. Amino acid substitutions in gene coding sequences are well documented as causes of variation in snake venom proteins, whereas the importance of gene regulation effects on venom protein abundance and composition is less well known. Here, we use a proteomics-based approach to infer the effects of gene regulation on protein expression by comparing the relative abundance of specific, known venom proteins among different individuals in each of two species of Sistrurus rattlesnakes. Variation in the presence or absence, and in the relative amounts, of proteins was high in both species across all major protein families. Based on our empirical criteria for inferring regulatory effects (presence-absence of specific proteins and/or more than threefold variation in abundance) between 51% and 83% of S. catenatus individuals and between 40% and 63% of S. miliarius individuals showed evidence for gene regulation across the four most abundant proteins (disintegrins, phospholipase A₂’s, serine proteinases, and snake venom metalloproteases). Thus, the effects of gene regulation should be considered an important cause of variation in the composition of whole venoms at the intraspecific level. They also suggest the need for testing the adaptive hypothesis for venom plasticity in relation to prey consumed by adult snakes. Finally, the venom variability reported may have an impact in the treatment of bite victims, highlighting the necessity of using pooled venoms as a substrate for antivenom production.