Corn Snake Research Articles

Reptarenaviruses in apparently healthy snakes in an Australian zoological collection.

Inclusion body disease (IBD) is a disease of snakes with a global distribution and has recently been shown to be caused by reptarenaviruses. Testing for this group of viruses in asymptomatic snakes allows the association between infection and disease to be further elucidated. A reptarenavirus was detected by RT-PCR in a reticulated python (Malayopython reticulatus) from an Australian zoological collection that was open-mouth breathing and had erythematous oral mucosa. Another 27 pythons, 4 elapids, 2 colubrids and 2 boas from this collection were then screened. From these animals, swabs, whole blood and/or tissue were tested for reptarenaviruses by RT-PCR. Additionally, blood films from 10 snakes were examined by light microscopy for the presence of inclusion bodies. The majority of samples were collected over a 484-day period. A total of 8 animals were RT-PCR-positive (8/36 = 22.2%): 6 were pythons, 1 was a corn snake (Pantherophis guttatus) and 1 was a Madagascar tree boa (Sanzinia madagascariensis). From them, 57 samples were collected, but only one from each animal was RT-PCR-positive (8/57 = 14.0%). From all 36 animals in this study, 8/182 samples were RT-PCR-positive (4.4%). Inclusion bodies were not recognised in any of the blood films. Only the reticulated python showed signs of illness, which improved without any further intervention. All other RT-PCR-positive snakes were apparently healthy throughout the duration of the study. This study showed a weak association between the presence of reptarenaviruses and disease. Testing serially collected swab and whole-blood samples increased the number of animals in which reptarenaviruses were detected.

Determination of the Feeding Schedule to Achieve Growth Optimization in Juvenile Snakes: A Case Study

Feeding schedules are an important aspect of any husbandry regime for animals in human care. While frequently debated for endothermic animals, ectotherms, especially snakes, often suffer from stagnated animal husbandry practices. To rectify this issue, we investigated the impact of food presentation frequency, as well as food amount per presentation on the growth of one-year-old and newborn Corn Snakes (Pantherophis guttatus) and newborn False Water Cobras (Hydrodynastes gigas) in human care. Three cohorts, each composed of siblings, were split into two treatment groups for the experiment. Each snake was fed two prey items representing less than 5% of their mass, approximately three days apart over a 15 week period. All animals were weighed weekly in grams. Treatment 1 received both prey items on the same day while treatment 2 received one prey item a day for 2 consecutive days. Over a 7 day period each individual consumed the same amount of food. The goal of this study was to test the impact of energy allocation on digestion and to determine if the digestion of a single large meal resulted in a loss of energy allocation for growth. Results indicated that there was not a major difference in growth between treatments, suggesting that energy allocated for digesting large amounts of prey did not result in a loss of energy allocation for growth.

The Effects of Temperature on the Turnover of δ18O and δD in Juvenile Corn Snakes (Elaphe guttata): A Novel Study with Ecological Implications

The use of natural variation in stable isotope ratios continues to be used in ecological studies without proper validation through laboratory studies. This study tested the effects of temperature, time, and turnover in the scales of juvenile corn snakes (Elaphe guttata) in a controlled, laboratory environment. Snakes were assigned to four treatment groups (24 °C, 27 °C, 30 °C, and freely thermoregulating), and one snake from each group was sacrificed weekly. Scales from each snake were washed, dried, and analyzed for δD and δ18O at the Stable Isotope Research Facility for Environmental Research at the University of Utah. The effects of temperature on the turnover of tissues was only significant when comparing the thermoregulating group to the pooled treatment groups (24 °C, 27 °C, and 30 °C) in the δ18O of scales (p = 0.006). After normalizing data on the δD and δ18O using percent change for comparison, δ18O appeared to be turning over at a faster rate than δD as indicated by an analysis of covariance (ANCOVA) test for homogeneity of slopes (F1,53 = 69.7, p < 0.001). With further testing of assumptions, a modification of our methods could provide information on the composition of drinking water sources in a species that switches between two isotopically distinct sources, such as during seasonal shifts in habitat or migration, and/or estimates of long-term field metabolic rates based on the turnover of these isotopes.

Making the best of a bad situation: differential predator avoidance in a diminutive woodland salamander

Predator avoidance behaviour has been observed in many species and can strongly affect trophic interactions. Animals frequently adjust their response to predators in proportion to risk; however, when they are exposed to chronic predation risk, some animals reduce their antipredator responses in favour of resource acquisition. Animals that are subjected to predation pressure from multiple predator species may have to respond to different spatial and temporal distributions of risk; how they balance these risks will affect their choice of habitat or microhabitat patches. One species that is exposed to a wide range of predators is the pygmy salamander, Desmognathus wrighti. These miniaturized terrestrial plethodontids spend their entire life on the forest floor where they are exposed to a diversity of predators. We examined pygmy salamanders' ability to avoid potential predators based on the presence of substrate-borne chemical cues. The predator species included ground beetles (Carabidae: Pterostichus), Jordan's salamanders, Plethodon jordani, and ring-necked snakes, Diadophis punctatus. The ring-necked snake is a trophic specialist feeding on salamanders while the beetles and Jordan's salamanders are generalist predators. The pygmy salamanders were also exposed to chemical alarm cues from a congeneric salamander (Desmognathus fuscus) and odours from two unfamiliar species: corn snakes (Pantherophis gutattus) and herbivorous desert tortoises (Gopherus agassizii). We used a hierarchical Bayesian beta regression with a repeated measures design to model proportion of time spent in each treatment. Compared to controls, pygmy salamanders exhibited strong aversion to fed ring-necked snakes, a moderate avoidance of unfamiliar odours and a slight attraction to scents from the two common generalist predators. Our results suggest that pygmy salamanders accept modest risk from common generalist predators but avoid less common, specialist predators and unfamiliar species.

Facultative mobilization of eggshell calcium promotes embryonic growth in an oviparous snake.

The mineralized eggshell of Reptilia was a major innovation in the evolution of the amniotic egg. Inorganic components strengthen the eggshell and are a potential source of nutrients to developing embryos. Embryos of oviparous reptiles do extract calcium from eggshells but vary interspecifically in exploitation of this resource. The pattern of embryonic calcium nutrition of the corn snake, Pantherophis guttatus, is similar to a diversity of squamate species: embryos obtain most calcium from yolk, yet also mobilize calcium from the eggshell. We tested the hypothesis that embryonic development is not dependent on eggshell calcium by manipulating calcium availability. We peeled away the outer calcareous layer of the eggshell of recently oviposited eggs; control eggs were left intact. Eggs were sampled periodically and calcium content of egg compartments (embryo, yolk, eggshell) was measured. We also analyzed skeletal development and size of hatchlings. There was no difference in survivorship or length of incubation between treatments. However, hatchlings from intact eggs contained more calcium and were larger in mass and length than siblings from peeled eggs. There were no observable differences in ossification but hatchlings from intact eggs had larger skeletal elements (skull, vertebrae). Our results indicate that mobilization of eggshell calcium is not a requirement for embryonic development of P. guttatus and that embryos augment yolk calcium by extracting calcium from the eggshell. This pattern of embryonic calcium nutrition would favor embryos with a greater capacity to mobilize calcium from the eggshell by promoting growth and thereby potentially enhancing hatchling fitness.

Reflections on Model Organisms in Evolutionary Developmental Biology.

This chapter reflects on and makes explicit the distinctiveness of reasoning practices associated with model organisms in the context of evolutionary developmental research. Model organisms in evo-devo instantiate a unique synthesis of model systems strategies from developmental biology and comparative strategies from evolutionary biology that negotiate a tension between developmental conservation and evolutionary change to address scientific questions about the evolution of development and the developmental basis of evolutionary change. We review different categories of model systems that have been advanced to understand practices found in the life sciences in order to comprehend how evo-devo model organisms instantiate this synthesis in the context of three examples: the starlet sea anemone and the evolution of bilateral symmetry, leeches and the origins of segmentation in bilaterians, and the corn snake to understand major evolutionary change in axial and appendicular morphology.

HNK-1 in Morphological Study of Development of the Cardiac Conduction System in Selected Groups of Sauropsida.

Human natural killer (HNK)-1 antibody is an established marker of developing cardiac conduction system (CCS) in birds and mammals. In our search for the evolutionary origin of the CCS, we tested this antibody in a variety of sauropsid species (Crocodylus niloticus, Varanus indicus, Pogona vitticeps, Pantherophis guttatus, Eublepharis macularius, Gallus gallus, and Coturnix japonica). Hearts of different species were collected at various stages of embryonic development and studied to map immunoreactivity in cardiac tissues. We performed detection on alternating serial paraffin sections using immunohistochemistry for smooth muscle actin or sarcomeric actin as myocardial markers, and HNK-1 to visualize overall staining pattern and then positivity in specific myocyte populations. We observed HNK-1 expression of various intensity distributed in the extracellular matrix and mesenchymal cell surface of cardiac cushions in most of the examined hearts. Strong staining was found in the cardiac nerve fibers and ganglia in all species. The myocardium of the sinus venosus and the atrioventricular canal exhibited transitory patterns of expression. In the Pogona and Crocodylus hearts, as well as in the Gallus and Coturnix ones, additional expression was detected in a subset of myocytes of the (inter)ventricular septum. These results support the use of HNK-1 as a conserved marker of the CCS and suggest that there is a rudimentary CCS present in developing reptilian hearts. Anat Rec, 302:69-82, 2019. © 2018 Wiley Periodicals, Inc.

Mice exposure to haloxyfop-p-methyl ester at predicted environmentally relevant concentrations leads to anti-predatory response deficit.

Although the efficiency of haloxyfop-p-methyl ester (HPME) as selective herbicide is acknowledged, its impact on non-target organisms is poorly known. It is not known whether the short exposure of mammals to low HPME concentrations (consistent with a realistic contamination scenario) poses risks to these animals. Thus, the aim of the current study is to evaluate the effects of HPME on the anti-predatory behavior of female Swiss mice exposed to it. The animals were divided in groups: non-exposed (control) and exposed (route: i.p., for 2days) to different herbicide concentrations (2.7 × 10-4g/kg and 2.7 × 10-2g/kg of body weight), which were considered environmentally relevant predicted concentrations. The animals were subjected to the open field and elevated plus-maze tests; results showed that the HPME did not lead to anxiolytic or anxiety behavior, or to locomotive changes in the tested animals, fact that was confirmed through the Basso Mouse Scale for locomotion scores. On the other hand, animals exposed to the herbicide were incapable of recognizing the snake as potential predator. Animals in the control group, exposed to a real snake (Pantherophis guttatus) remained longer in the safety zone of the test device, presented lower frequency of self-grooming behaviors for a shorter period-of-time, besides showing longer freezing time, which was not observed in animals exposed to HPME. Therefore, our study indicates the ecotoxicological potential of the herbicide, since anti-predatory behavior disorders may affect preys' responses and population dynamics.

The effects of predicted environmentally relevant concentrations of ZnO nanoparticles on the behavior of Gallus gallus domesticus (Phasianidae) chicks

The toxicity of ZnO nanoparticles (NPs) has been the subject of several investigations; however, concentrations much higher than the ones potentially found in the environment are often tested. In addition, groups of animals such as birds have not been used as model in studies in this field, fact that creates an important ecotoxicological gap in them. The aim of the present study is to investigate the effects of the exposure to environmentally relevant concentrations of ZnO nanoparticles on the anti-predatory behavior of chicks (Gallus gallus domesticus). The test animals were daily exposed to an environmentally relevant concentration of ZnO nanoparticles (0.245 mg kg−1) and to a toxic concentration of it (245.26 mg kg−1) through intraperitoneal injection for two days. We set a control group for comparison purposes. According to our results, ZnO nanoparticles did not affect the locomotor activity of, and did not cause anxiolytic or anxiogenic effect on, birds in the open field test. However, based on the lowest cluster score recorded during the social aggregation test, chicks exposed to ZnO nanoparticles failed to recognize the grunt of a hawk (Rupornis magnirostris) as predatory threat. Only birds in the control group recognized the test snake (Pantherophis guttatus) as potential predator. The higher Zn concentration in the brains of animals exposed to ZnO nanoparticles evidenced the capacity of these nanomaterials to cross the blood–brain barrier, even at low concentrations. This blood-brain barrier crossing could have affected the structures or neuronal mechanisms that modulate the defensive response of birds. Assumingly, even the minimal exposure to low concentrations of ZnO nanoparticles can affect birds. Our outcomes corroborate previous studies about the biological risks of water surface contamination by metal-based nanomaterials.

Electrical field stimulation-induced contractions on Pantherophis guttatus corpora cavernosa and aortae.

A tetrodotoxin (TTX)-resistant mechanism is responsible for the electrical field stimulation (EFS)-induced contractions and relaxations of Crotalus durissus terrificus corpora cavernosa. Here it was investigated whether this mechanism also occurs in corpora cavernosa and aortae of the non-venomous snake Pantherophis guttatus corpora cavernosa and aortae. Corpora cavernosa and aortic rings isolated from Pantherophis guttatus snake were mounted in organ bath system for isometric tension recording. EFS-induced contractions in both tissues were performed in the presence and absence of guanethidine (30 μM), phentolamine (10 μM) and tetrodotoxin (1 μM). In another set of experiments, the endothelium was removed from aortic rings and EFS-induced contractions were performed in the denuded rings. Electrical field stimulation-induced contractions were frequency-dependent in Pantherophis guttatus corpora cavernosa and aortic rings. The contractions were significantly reduced in the presence of guanethidine (30 μM) or phentolamine (10 μM). Pre-treatment with tetrodotoxin had no effect on the EFS-induced contractions of either corpora cavernosa or aortic rings. Surprisingly, the EFS-induced contractions of aortic rings denuded of endothelium were almost abolished. These results indicate that the TTX-resistant mechanism is present in EFS-induced contractions of Pantherophis guttatus corpora cavernosa and aortae. The experiments performed in the aorta indicate that the endothelium is the main source for the release of catecholamines induced by EFS.

The intake of water containing a mix of pollutants at environmentally relevant concentrations leads to defensive response deficit in male C57Bl/6J mice

Previous studies have individually confirmed the toxic effects from different pollutants on mammals. However, effects resulting from the exposure of these animals to multi-pollutant mixes have not been studied so far. Thus, the aim of the current study is to assess the effect from the chronic exposure (105days) of C57Bl/6J mice to a mix of pollutants on their response to potential predators. In order to do so, the following groups were formed: “control”, “Mix 1× [compounds from 15 pollutants identified in surface waters at environmentally relevant concentration (ERC)]”, “Mix 10×” and “Mix 25×” (concentrations 10 and 25 times higher than the ERC). From the 100th experimental day on, the animals were subjected to tests in order to investigate whether they showed locomotor, visual, olfactory and auditory changes, since these abilities are essential to their anti-predatory behavior. Next, the animals' behavior towards potential predators (Felis catus and Pantherophis guttatus) was assessed. The herein collected data did not show defensive response from any of the experimental groups to the predatory stimulus provided by P. guttatus. However, the control animals, only, presented anti-predatory behavior when F. catus was introduced in the apparatus, fact that suggests defensive response deficit resulting from the treatments. Thus, the current study is pioneer in showing that the chronic intake of water containing a mix of pollutants (even at low concentrations) leads to behavioral disorders able to affect the survival and population dynamics of mammalian species at ecological level.

METHOD OF SEMEN COLLECTION AND ARTIFICIAL INSEMINATION IN SNAKES

This study focuses on the method for sperm extraction and artificial insemination in snakes. Ten adult healthy snakes (4.6) were included in the study (Pantherophis guttatus 1.3; Hydrodynastes gigas 1.1; Corallus hortulanus 1.1; and Sanzinia madagascariesis 1.1). Massage of the ventral aspect of the caudal third of the male snake body for 2 to 3 minutes was performed successfully for the semen collection. Both female and male P. guttatus and H. gigas were placed in brumation, with the female snakes being assessed for ovarian activity after emerging from dormancy. Only females showing ultrasonographic evidence of vitellogenic follicles were included in the study. S. madagascariensis and C. hortulanus were maintained at the same temperature through the year, and the ovarian activity was assessed ultrasonographically prior to artificially inseminating the animal. With the aid of a rigid endoscope, fresh semen was delivered through the cloaca and into the females’ oviducts using a catheter connected to the syringe. The technique failed in female S. madagascariensis and H. gigas. Two female P. guttatus laid eggs 2 months after artificial insemination, with hatchlings emerging following 2 months of development within the eggs. The third female corn snake did not lay an egg. The female C. hortulans produced eggs 4 months after insemination. The manual massage method of sperm collection in male snakes and endoscopy-assisted insemination in female snakes may be useful for conservation programs.

Terrestrial Snake Environmental DNA Accumulation and Degradation Dynamics and its Environmental Application

Abstract There is an increasing need for effective biomonitoring tools that quantify patterns of habitat occupancy by reptile species. Environmental DNA (eDNA) has been regarded as an emerging tool to detect specific target species; however, the dynamics of accumulation and degradation of eDNA in terrestrial environments are poorly understood. This study determines the time required for terrestrial snakes to leave enough eDNA behind to become detectable (accumulation time) as well as its persistence (degradation time). By targeting mitochondrial cytochrome oxidase subunit I and 12S rRNA genes of Red Cornsnakes (Pantherophis guttatus) in a controlled laboratory setting, we found that eDNA can be detected 3.5 h after the snakes had contact with soil and for up to 6 d after their removal. Estimated accumulation rate of Pantherophis guttatus eDNA per gram of snake biomass per hour was 12.6 μg. We also evaluated the applicability of eDNA detection under field conditions by targeting the mitochondrial cytochrom...

Relative position of the atrioventricular canal determines the electrical activation of developing reptile ventricles.

Squamate reptiles appear to lack the specialized His-Purkinje system that enables the cardiac ventricle to be activated from apex to base as in mammals and birds. Instead, activation may simply spread from where the atrioventricular canal connects to the base. Gja5, which encodes Cx40, which allows fast impulse propagation, was expressed throughout the ventricles of developing anole lizards. Activation was optically recorded in developing corn snake and central bearded dragon. Early embryonic ventricles were broad in shape, and activation propagated from the base to the right. Elongated ventricles of later stages were activated from base to apex. Before hatching of the snake, the ventricle developed a cranial extension on the left and activation propagated from the base to the caudal apex and the cranial extension. In squamate reptiles, the pattern of electrical activation of the cardiac ventricle is dependent on the position of the atrioventricular canal and the shape of the ventricle.

OCA2 splice site variant in German Spitz dogs with oculocutaneous albinism.

We investigated a German Spitz family where the mating of a black male to a white female had yielded three puppies with an unexpected light brown coat color, lightly pigmented lips and noses, and blue eyes. Combined linkage and homozygosity analysis based on a fully penetrant monogenic autosomal recessive mode of inheritance identified a critical interval of 15 Mb on chromosome 3. We obtained whole genome sequence data from one affected dog, three wolves, and 188 control dogs. Filtering for private variants revealed a single variant with predicted high impact in the critical interval in LOC100855460 (XM_005618224.1:c.377+2T>G LT844587.1:c.-45+2T>G). The variant perfectly co-segregated with the phenotype in the family. We genotyped 181 control dogs with normal pigmentation from diverse breeds including 22 unrelated German Spitz dogs, which were all homozygous wildtype. Comparative sequence analyses revealed that LOC100855460 actually represents the 5’-end of the canine OCA2 gene. The CanFam 3.1 reference genome assembly is incorrect and separates the first two exons from the remaining exons of the OCA2 gene. We amplified a canine OCA2 cDNA fragment by RT-PCR and determined the correct full-length mRNA sequence (LT844587.1). Variants in the OCA2 gene cause oculocutaneous albinism type 2 (OCA2) in humans, pink-eyed dilution in mice, and similar phenotypes in corn snakes, medaka and Mexican cave tetra fish. We therefore conclude that the observed oculocutaneous albinism in German Spitz is most likely caused by the identified variant in the 5’-splice site of the first intron of the canine OCA2 gene.

Reproductive Traits in the Great Plains Ratsnake, Pantherophis emoryi (Baird and Girard, 1853), in Northern Kansas

Museum specimens of the Great Plains Ratsnake (Pantherophis emoryi) from six counties in northern Kansas were examined to determine gonadal cycle, clutch characteristics, and body sizes at sexual maturity near the northern edge of its geographic range. Gonadal cycle of males of northern Kansas adhered to a temperate pattern unlike that of Eastern Corn Snakes (P. guttatus) of southern Florida, whereas females of both species from these locations maintained a tropical pattern to their monthly distribution of follicle size. Clutch size averaged 9.5 eggs in this study and 7.2 eggs in a study of this species in northeastern Kansas. Both of these values were smaller than those of southern and eastern populations of the Eastern Corn Snake despite larger or similar body sizes, respectively, of the latter group. We discuss the potential effects of climate change on male reproduction and clutch size.

Pet snakes illegally marketed in Brazil: Climatic viability and establishment risk.

Invasive species are one among many threats to biodiversity. Brazil has been spared, generically, of several destructive invasive species. Reports of invasive snakes’ populations are nonexistent, but the illegal pet trade might change this scenario. Despite the Brazilian laws forbid to import most animals, illegal trade is frequently observed and propagules are found in the wild. The high species richness within Brazilian biomes and accelerated fragmentation of natural reserves are a critical factors facilitating successful invasion. An efficient way to ease damages caused by invasive species is identifying potential invaders and consequent prevention of introductions. For the identification of potential invaders many factors need to be considered, including estimates of climate matching between areas (native vs. invaded). Ecological niche modelling has been widely used to predict potential areas for invasion and is an important tool for conservation biology. This study evaluates the potential geographical distribution and establishment risk of Lampropeltis getula (Linnaeus, 1766), Lampropeltis triangulum (Lacépède, 1789), Pantherophis guttatus (Linnaeus, 1766), Python bivittatus Kuhl, 1820 and Python regius (Shaw, 1802) through the Maximum Entropy modelling approach to estimate the potential distribution of the species within Brazil and qualitative evaluation of specific biological attributes. Our results suggest that the North and Midwest regions harbor major suitable areas. Furthermore, P. bivittatus and P. guttatus were suggested to have the highest invasive potential among the analyzed species. Potentially suitable areas for these species were predicted within areas which are highly relevant for Brazilian biodiversity, including several conservation units. Therefore, these areas require special attention and preventive measures should be adopted.

A Simulated Heat Wave Has Diverse Effects on Immune Function and Oxidative Physiology in the Corn Snake (Pantherophis guttatus).

Animals will continue to encounter increasingly warm environments, including more frequent and intense heat waves. Yet the physiological consequences of heat waves remain equivocal, potentially because of variation in adaptive plasticity (reversible acclimation) and/or aspects of experimental design. Thus, we measured a suite of physiological variables in the corn snake (Pantherophis guttatus) after exposure to field-parameterized, fluctuating temperature regimes (moderate temperature and heat wave treatments) to address two hypotheses: (1) a heat wave causes physiological stress, and (2) thermal performance of immune function exhibits adaptive plasticity in response to a heat wave. We found little support for our first hypothesis because a simulated heat wave had a negative effect on body mass, but it also reduced oxidative damage and did not affect peak performance of three immune metrics. Likewise, we found only partial support for our second hypothesis. After exposure to a simulated heat wave, P. guttatus exhibited greater performance breadth and reduced temperature specialization (the standardized difference between peak performance and performance breadth) for only one of three immune metrics and did so in a sex-dependent manner. Further, a simulated heat wave did not elicit greater performance of any immune metric at higher temperatures. Yet a heat wave likely reduced innate immune function in P. guttatus because each metric of innate immune performance in this species (as in most vertebrates) was lower at elevated temperatures. Together with previous research, our study indicates that a heat wave may have complex, modest, and even positive physiological effects in some taxa.

A Novel Pattern of Yolk Processing in Developing Snake Eggs (Colubridae: Lampropeltini) and its Functional and Evolutionary Implications.

Early amniotic vertebrates evolved large-yolked eggs that permitted production of well-developed, terrestrial hatchlings. This reproductive pattern required new mechanisms for cellularizing the yolk and mobilizing it for embryonic use. In birds, cells that line the yolk sac cavity phagocytose and digest the yolk material, a pattern that is commonly assumed to be universal among oviparous amniotes. However, recent evidence challenges the assumption that all squamate reptiles conform to the avian developmental pattern. In this paper, scanning electron microscopy and histology were used to study mechanisms of yolk processing in two colubrid snakes, the kingsnake Lampropeltis getula and the milksnake L. triangulum. Endodermal cells from the yolk sac splanchnopleure proliferate massively as they invade the yolk sac cavity, forming elaborate chains of interlinked cells. These cells grow in size as they phagocytose yolk material. Subsequently, vitelline capillaries invade the masses of yolk-laden cells and become coated with the endodermal cells, forming an elaborate meshwork of cell-coated strands. The close association of cells, yolk, and blood vessels allows yolk material to be cellularized, digested, and transported for embryonic use. The overall pattern is like that of the corn snake Pantherophis guttatus, but contrasts markedly with that of birds. Given recent evidence that this developmental pattern may also occur in certain lizards, we postulate that it is ancestral for squamates. Studies of lizards, crocodilians, and turtles are needed to clarify the evolutionary history of this pattern and its implications for the evolution of the amniotic (terrestrial) vertebrate egg.

Immunologic responses in corn snakes (Pantherophis guttatus) after experimentally induced infection with ferlaviruses.

OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses. ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes. PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days -6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed. RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses. CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.