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PubMed2003

Hemorrhagic activity of the Duvernoy's gland secretion of the xenodontine colubrid Philodryas patagoniensis from the north-east region of Argentina.

Colubrid snakes belonging to Philodryas genus, widespread all over South America, bring about lesions (swelling, ecchymosis, transient bleeding from the bite site punctures), that are similar to those produced by Bothrops species (yarará). In the present work we began the characterization of Philodryas patagoniensis venom. We examined if this venom produces hemorrhagic lesions as those observed in victims bitten by Philodryas olfersii. Hemorrhagic, proteolytic and fibrinogenolytic activities were evaluated, and histological observations in samples of gastrocnemius muscle were carried out. Inhibition studies were carried out in metal chelator (ethylenediaminetetraacetic acid) presence. Our results show a small Minimum Hemorrhagic Dose (MHD=0.035 microg) and a high proteolytic activity (143 U/mg), and prove the capacity of this venom to degrade fibrinogen in vitro rendering it unclottable by thrombin, supporting the presence of proteases, principally metalloproteases, in P. patagoniensis venom that are able to alterate the vascular wall and degrade fibrinogen, being both activities responsible of a high hemorrhagic activity.

Animals↗
PubMed1997

Venom yields from several species of colubrid snakes and differential effects of ketamine.

The composition of rear-fanged colubrid snake venoms is largely unknown due primarily to the difficulty involved in venom collection. Several different methods have been used to maximize the yield of Duvernoy's secretions. The method proposed by Rosenberg in 1992, which includes the use of ketamine hydrochloride anesthetic and pilocarpine to induce Duvernoy's glands secretion, was used in the present study to collect venom from eight species of colubrids. Protein concentrations, using a dye-binding microassay technique, were determined for the venoms collected. Average protein concentrations ranged from 49.8 to 96.4%. Most yields (dry weight/snake) obtained from specimens in this study were significantly greater than yields previously reported. There was a wide range of effects that occurred due to the ketamine injections; however, all snakes recovered from the effects of the ketamine hydrochloride/pilocarpine with no apparent ill effects. Recommended doses of ketamine hydrochloride have thus been adjusted, depending on previous reactions to the drug. The use of ketamine/pilocarpine in the collection of Duvernoy's secretion has proven to be highly effective in increasing yields. Some caution should be observed when administering ketamine to various species of colubrids, as effects do not necessarily scale to body mass.

Anesthetics, Dissociative↗
PubMed1997

Enzymatic properties of the Duvernoy's secretion of Blanding's tree snake (Boiga blandingi) and of the mangrove snake (Boiga dendrophila).

Duvernoy's secretion from representatives of the colubrid genus Boiga has been shown by other workers: (1) to contain an alpha-bungarotoxin-like component (Blanding's tree snake, Boiga blandingi); (2) to exhibit acetylcholine receptor binding activity (the brown tree snake, Boiga irregularis); and (3) to react strongly (B. blandingi and the mangrove snake, Boiga dendrophila) with antisera raised against elapid venoms. The present study provides the first evidence for the presence of acetylcholinesterase, a common elapid enzyme, in the Duvernoy's secretion of B. dendrophila and B. blandingi at levels of 122 +/- 7 and 0.36 +/- 0.02 International Enzyme Units, respectively. Constants for inhibition by eserine were 1.25 x 10(-7) M and 2.79 x 10(-7) M, respectively. The Duvernoy's secretion of B. dendrophila exhibited significant phospholipase and haemolytic activities, whereas phosphodiesterase was only weakly present and no thrombin-like activity was observed.

Acetylcholinesterase↗
PubMed1998

Characterization of a myotoxin from the Duvernoy's gland secretion of the xenodontine colubrid Philodryas olfersii (green snake): effects on striated muscle and the neuromuscular junction.

A myotoxin has been isolated from the Duvernoy's gland (DG) secretion of the xenodontine colubrid Philodrvas olfersii (green snake) by gel filtration on Sephadex G-100 SF. Under non-reducing and reducing conditions in SDS-PAGE, the myotoxin migrates as a single band with a mol. wt. of 20000. The toxin has 182 amino acid residues (approximately 20% acidic), a pI of 4.8 and a blocked N-terminal. In the chick biventer cervicis preparation, P. olfersii myotoxin partially blocks potassium-evoked contractures without affecting either the twitch-tension resulting from indirect stimulation or the contractures evoked by acetylcholine. Both the DG secretion and the myotoxin increase the serum creatine kinase (CK) levels of mice and stimulate the release of CK from the biventer cervicis preparation in a dose- and time-dependent manner. The varying degrees of muscle cell lysis and extensive widening of the intercellular spaces caused by the DG secretion are reproduced by the myotoxin, with the exception that in the latter the partial or total loss of transverse muscle striations is restricted to the muscle periphery. This myotoxin is the first such protein to be characterized from a DG secretion.

Acetylcholine↗
PubMed2004

Trophic and maternal transfer of selenium in brown house snakes (Lamprophis fuliginosus).

Excessive concentrations of dietary Se are toxic to oviparous vertebrates (i.e., fish and birds) but little is known about its accumulation and effects in reptiles. We exposed female brown house snakes, Lamprophis fuliginosus, to 10 and 20 microg/g Se by injecting seleno-D,L-methionine into their prey items and compared the snakes to individuals receiving background levels of approximately 1 microg/g dietary Se. Snakes were fed meals equaling 25% of their body mass 2-3 times a month for 10 months. Snakes exposed to excessive Se accumulated significant concentrations of Se in kidney, liver, and ovarian tissue, but accumulation had no effect on female survival, food consumption, growth, or body condition. Fewer females exposed to excessive Se reproduced than females exposed to 1 microg/g Se (67% vs. 91%, respectively), but the reduction in reproductive activity was not statistically significant. Total reproductive output of females did not differ among the three dietary treatments. However, snakes exposed to 10 and 20 microg/g Se transferred significant concentrations of Se to their eggs. In the 20 microg/g treatment, maternal transfer resulted in Se concentrations in eggs that surpassed all suggested reproductive toxicity thresholds for birds and fish. Further studies are needed to more rigorously determine whether maternal transfer of Se in this snake species affects the viability of developing embryos or the health of offspring.

Animals↗
PubMed1999

Signal transduction in the vomeronasal organ of garter snakes: ligand-receptor binding-mediated protein phosphorylation.

The vomeronasal (VN) system of garter snakes plays an important role in several species-typical behaviors, such as prey recognition and responding to courtship pheromones. We (X.C. Jiang et al., J. Biol. Chem. 265 (1990) 8736-8744 and Y. Luo et al., J. Biol. Chem. 269 (1994) 16867-16877) have demonstrated previously that in the snake VN sensory epithelium, the chemoattractant ES20, a 20-kDa glycoprotein derived from electric shock-induced earthworm secretion, binds to its receptor which is coupled to PTX-sensitive G-proteins. Such binding results in elevated levels of IP3. We now report that ES20-receptor binding regulates the phosphorylation of two membrane-bound proteins with molecular masses of 42- and 44-kDa (p42/44) in both intact and cell-free preparations of the VN sensory epithelium. ES20 and DAG regulate the phosphorylation of p42/44 in a similar manner. ES20-receptor binding-mediated phosphorylation of p42/44 is rapid and transient, reaching a peak value within 40 seconds and decaying thereafter. Phosphorylation of p42/44 appears to be regulated by the countervailing actions of a specific membrane-bound protein kinase and a protein phosphatase. The phosphorylation of these membrane-bound proteins significantly reduces the activity of G-proteins as evidenced by a decrease in GTPase activity, but has little effect on ligand-receptor binding. These findings suggest that p42/44 play a role in modulating the signal transduction induced by ES20 in the vomeronasal system.

Animals↗
PubMed1999

Isolation and experimental transmission of a reovirus pathogenic in ratsnakes (Elaphe species).

A reovirus was isolated from juvenile Moellendorff's ratsnakes (Elaphe moellendorffi) and beauty snakes (Elaphe taenuris) that died soon after importation into the USA. Viper heart (VH2) cells inoculated with tissue homogenates showed cytopathic effects consisting of large syncytia formation followed by cell detachment from the monolayer. Tissue culture supernatants failed to hemagglutinate guinea pig and chicken erythrocytes at room temperature. Electron microscopy of purified virions revealed spherical to icosahedral particles measuring 70-85 nm in diameter with a double capsid layer. Preparations of the viral genome contained ten segments of dsRNA when analyzed by polyacrylamide gel electrophoresis. A juvenile black ratsnake (Elaphe obsoleta obsoleta) was experimentally inoculated with the isolate and was found dead 26 days post inoculation. Necropsy revealed diffuse subacute interstitial pneumonia with respiratory epithelial cell hyperplasia and syncytia. Reovirus isolated from this snake was used to inoculate another juvenile black ratsnake which was euthanized 40 days post inoculation. Pneumonia and multifocal subacute proliferative tracheitis were found on necropsy. Reovirus was isolated from the lung of this snake and was demonstrated by transmission electron microscopy. This is the first documentation of a pathogenic reptile reovirus and the first report of experimental transmission of a reovirus in snakes.

Animals↗
PubMed1994

Human envenomation from a wandering garter snake.

Garter snake bites are generally innocuous to human beings. We report a case of human envenomation from the Wandering Garter snake (Thamnophis elegans vagrans). The patient, who was bitten on his right third fingertip, rapidly developed local edema, ecchymosis, and hemorrhagic vesicles. Systemic signs and symptoms did not develop. The clinical picture was similar to that in three previous patients with Thamnophis envenomation in that clinical signs followed a prolonged bite. Thamnophis species have Duvernoy's glands, which may be analogous to venom glands in Crotalidae (pit viper) species. The progressive local effects produced by secretions of these glands may be confused with early Crotalidae envenomation.

Adolescent↗
PubMed2002

Distribution of aromatase immunoreactivity in the forebrain of red-sided garter snakes at the beginning of the winter dormancy.

Until recently, it has been difficult to identify the exact location of aromatase containing cells in the brain. The development of new antibodies has provided a sensitive tool to analyze the distribution of aromatase immunoreactive (ARO-ir) material at a cellular level of resolution. In the present study we examined, for the first time, the distribution of ARO-ir cells in the brain of a reptile, the red-sided garter snake, at the beginning of the winter dormancy. ARO-ir cells were found at all rostro-caudal levels in the red-sided garter snake brain. Although weakly stained cells were distributed throughout the brain, more intensely immunoreactive cells were primarily concentrated in the preoptic area, anterior hypothalamus, septum and nucleus sphericus. Although androgens are elevated upon emergence from hibernation in the male red-sided garter snake, initiation of courtship behavior appears to be independent of direct androgen control. To date, the only known stimulus found to initiate courtship is a period of low temperature dormancy followed by exposure to warm temperatures. Circumstantial data, however, suggest an indirect role in the activation of male copulatory behavior for estrogenic metabolites of testosterone produced in the brain by aromatization during the winter dormancy. This study provides the first documentation of the distribution of ARO-ir cells in a reptilian species and demonstrates that while the aromatase enzyme occurs in most regions of the brain, the ARO-ir cells that appear to contain the highest concentration of enzyme are clustered in brain areas classically associated with the control of courtship behavior and mating in vertebrates. These data are consistent with the idea that estrogens locally produced in the brain may participate in some way to the activation of sexual behavior in this species also. This notion should now be experimentally tested by analyzing annual changes in aromatase activity and immunoreactivity and assessing the effects of pharmacological blockade of the enzyme activity at different times of the year.

Animals↗
PubMedDate not supplied

Differential vulnerability of snake species to MPTP: a behavioral and biochemical comparison in ratsnakes (Elaphe) and watersnakes (Nerodia).

The synthetic neurotoxicant 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induces a Parkinsonian-like syndrome in humans and nonhuman primates, and also causes movement disorders in rodents, fish, amphibians and lizards. To date, the effects of MPTP have not been characterized in snakes. In this study, the behavioral and biochemical effects of MPTP were assessed in the black ratsnake Elaphe o. obsoleta and the banded watersnake Nerodia f. fasciata--species that display contrasting behavioral sensitivities to dopaminergic antagonists and to amphibian toxins. We report that MPTP induces depletion of norepinephrine and serotonin in fore, mid and hindbrain regions and depletion of dopamine in fore and midbrain regions in E.o. obsoleta. MPTP also induced a marked reduction in righting ability in E.o. obsoleta. In N.f. fasciata, norepinephrine and dopamine were depleted by MPTP in all three brain regions and serotonin was only significantly reduced in the forebrain. In contrast to E.o. obsoleta, N.f. fasciata demonstrated no behavioral disorders. This study demonstrates a behavioral and biochemical sensitivity to MPTP in E.o. obsoleta that differs from that in N.f. fasciata. The differential sensitivities to monoaminergic modulation may be related to the contrasting diets of these species.

Animals↗
PubMed1994

An ultrastructural study of the dorsal lingual epithelium of the rat snake, Elaphe quadrivirgata.

The histological characteristics and ultrastructure of the dorsal lingual epithelium of the rat snake, Elaphe quadrivirgata, were investigated by light microscopy and scanning and transmission electron microscopy. Most of the surface of the bifurcated part of the tongue was relatively smooth. Dome-shaped, hemispherical bulges were compactly arranged on the epithelial cell surface of the basal area of this region. Intercellular borders were clearly recognizable as striations. Microridges were densely distributed on the epithelial cell surface of the lingual body. Intercellular borders were thickened. A keratinized layer was clearly visible in the epithelium of the anterior bifurcated area, namely, at the apex of the tongue. Although keratohyalin granules were not found in any layer of the epithelium in this area, the cells of the surface layer were filled with keratin filaments. The dorsal lingual epithelium of the posterior area, namely, the lingual body, did not show any evidence of keratinization. Each cell on the surface side still had a large, oval nucleus and intact organelles, such as mitochondria, rough endoplasmic reticulum, ribosomes, tonofibrils, and tonofilaments. Cellular interdigitation was evident between adjacent cells and clear microridges or microvilli were observed on the cell membranes on the free-surface side of cells located in the surface layer. The phylogenetic relevance of these findings is discussed.

Animals↗
PubMed2002

Phylogeographic analysis of the cornsnake (Elaphe guttata) complex as inferred from maximum likelihood and Bayesian analyses.

Most phylogeographic studies have used maximum likelihood or maximum parsimony to infer phylogeny and bootstrap analysis to evaluate support for trees. Recently, Bayesian methods using Marlov chain Monte Carlo to search tree space and simultaneously estimate tree support have become popular due to its fast search speed and ability to create a posterior distribution of parameters of interest. Here, I present a study that utilizes Bayesian methods to infer phylogenetic relationships of the cornsnake (Elaphe guttata) complex using cytochrome b sequences. Examination of the posterior probability distributions confirms the existence of three geographic lineages. Additionally, there is no support for the monophyly of the subspecies of E. guttata. Results suggest the three geographic lineages partially conform to the ranges of previously defined subspecies, although Shimodaira-Hasegawa tests suggest that subspecies-constrained trees produce significantly poorer likelihood estimates than the most likely trees reflecting the evolution of three geographic assemblages. Based on molecular support, these three geographic assemblages are recognized as species using evolutionary species criteria: E. guttata, Elaphe slowinskii, and Elaphe emoryi [phylogeographic, maximum likelihood, maximum parsimony, bootstrap, Bayesian, Markov chain Monte Carlo, cornsnake, Cytochrome b, geographic lineages, E. guttta, E. slowinskii, and E. emoryi].

Animals↗
PubMed2001

The effects of thermal and hydric environments on hatching success, embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata.

We examined the effects of thermal and hydric environments on hatching success, the embryonic use of energy and hatchling traits in a colubrid snake, Elaphe carinata. The eggs were incubated at four temperatures ranging from 24 to 32 degrees C on substrates with water potentials of 0 and -220 kPa using a 4x2 factorial design. Both thermal and hydric environments affected the water exchange between eggs and their surroundings. Eggs incubated in wetter substrates gained mass throughout the course of incubation, whereas eggs in drier substrates gained mass during the first half of incubation and lost mass thereafter. Hatching success was noticeably higher at 26 and 30 degrees C than at 24 and 32 degrees C, but among treatments, differences in hatching success were not significant. Temperature significantly affected the duration of incubation and most hatchling traits examined. Deformed hatchlings were found in all temperature treatments, with more deformities observed at 32 degrees C. Hatchlings from eggs incubated at different temperatures differed in wet body mass, but the differences stemmed mainly from variation in water contents. Embryos at different temperatures completed development at nearly the same expenditure of energy and catabolized nearly the same amount of lipids, but hatchlings from different temperatures differed in the development condition of carcass at hatching. Hatchlings from eggs incubated at 26 degrees C were larger in SVL than those from other higher or lower incubation temperatures, characteristically having larger carcasses; hatchlings from 32 degrees C eggs were smaller in SVL and had smaller carcasses but larger residual yolks than those from lower incubation temperatures. Hatchlings from eggs incubated at 24 degrees C were shorter in tail length but greater in size (SVL)-specific body wet mass than those from higher incubation temperatures. Within the range from -220 to 0 kPa, the substrate water potential did not affect hatching success, the embryonic use of energy and all hatchling traits examined, and the effects of temperature were independent of the effects of substrate water potential. Therefore, our data add evidence showing that embryonic development in reptiles with pliable-shelled eggs is relatively insensitive to variation in hydric environments during incubation.

Animals↗
PubMed2003

Non-invasive measure of body composition of snakes using dual-energy X-ray absorptiometry.

Non-invasive techniques to measure body composition are critical for longitudinal studies of energetics and life histories and for investigating the link between body condition and physiology. Previous attempts to determine, non-invasively, the body composition of snakes have proven problematic. Therefore, we explored whether dual-energy X-ray absorptiometry (DXA) could be used to determine the body composition of snakes. We analyzed 20 adult diamondback water snakes (Nerodia rhombifer) with a DXA instrument and subsequently quantified their body composition by gravimetric and chemical extraction methods. Body composition components scaled with body mass with mass exponents between 0.88 and 1.53. DXA values for lean tissue mass, fat mass and total-body bone mineral mass were significantly correlated with observed masses of lean tissue, fat and ash from chemical analysis. Using regression models incorporating DXA values we predicted the fat-free tissue mass, lean tissue mass, fat mass, ash mass and total body water content for this sample of water snakes. A cross-validation procedure demonstrated that these models estimated fat-free tissue mass, lean tissue mass, fat mass, ash mass and total-body water content with respective errors of 2.2%, 2.3%, 16.0%, 6.6% and 3.5%. Compared to other non-invasive techniques, include body condition indices, total body electrical conductivity (TOBEC) and cyclopropane absorption, DXA can more easily and accurately be used to determine the body composition of snakes.

Absorptiometry, Photon↗
PubMed1998

Definition of the specific roles of lysolecithin and palmitic acid in altering the susceptibility of dipalmitoylphosphatidylcholine bilayers to phospholipase A2.

Bilayers composed of phosphatidylcholine initially resist catalysis by phospholipase A2. However, after a latency period, they become susceptible when sufficient reaction products (lysolecithin and fatty acid) accumulate in the membrane. Temperature near the main bilayer phase transition and calcium concentration modulate the effectiveness of the reaction products. The purpose of this study was to examine the individual contributions of lysolecithin and palmitic acid to the susceptibility of dipalmitoylphosphatidylcholine vesicles and to rationalize the effects of temperature and calcium. Various fluorescent probes (Prodan, Laurdan, pyrene-labeled fatty acid, and dansyl-labeled phospholipid) were used to assess changes in the ability of the reaction products to perturb the bilayer and to affect the interactions with the enzyme. Un-ionized palmitic acid decreased bilayer polarity and perturbed the membrane surface exposing some of the Prodan to bulk water. Lysolecithin increased bilayer polarity and the rate of dipolar relaxation in response to the excited states of Laurdan and Prodan. A combination of the individual contributions of each product was observed when palmitic acid and lysolecithin were present together at low calcium, and the effects of lysolecithin dominated at high calcium. Palmitic acid, but not lysolecithin, promoted the binding of phospholipase A2 to the bilayer surface in the absence of calcium. Lysolecithin reduced the ability of fatty acid to enhance binding apparently by altering the structure of fatty acid domains in the membrane. Furthermore, increased temperature and ionization of the fatty acid tended to cause segregation of bound phospholipase A2 into domains poor in phospholipid content which presumably impeded bilayer hydrolysis. In contrast, un-ionized palmitic acid and lysolecithin promoted hydrolysis by augmenting a step distal to the adsorption of enzyme to the bilayer. This kinetic response to lysolecithin was calcium-dependent. A model accounting for these varied influences of the reaction products is presented.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2002

Risk assessment of an acetaminophen baiting program for chemical control of brown tree snakes on Guam: evaluation of baits, snake residues, and potential primary and secondary hazards.

The brown tree snake (Boiga irregularis) is a significant ecological, agricultural, and economic pest on Guam. Acetaminophen has recently been identified as a promising snake toxicant. Subsequent experimentation has shown that acetaminophen-mouse baits are readily consumed by and acutely toxic to browntree snakes. Before implementing an island-wide acetaminophen-mouse baiting program for the reducton of brown tree snake populations,the potential risks to nontarget wildlife must be evaluated. Quantification of nontarget hazards by comparing potential exposure levels to toxicity values suggested a significant level of concern for rodents, cats, pigs, and birds. For these species, subsequent calculations and field and laboratory experiments, which quantified acetaminophen consumption under field conditions, indicated that acetaminophen consumption was minimal. These results indicate that the advantages of using acetaminophen to reduce brown tree snake populations on Guam outweigh the minimal risks to nontarget feral and wildlife species.

Acetaminophen↗
PubMed2003

Laser ablation-ICP-MS analysis of dissected tissue: a conservation-minded approach to assessing contaminant exposure.

Minimally invasive sampling techniques are an essential ecotoxicological tool for continuous assessment of contaminant exposure and in instances where it is not desirable or practical to sacrifice the animal. In this paper, we report on the application of laser ablation-ICP-MS (LA-ICP-MS) for sampling of minute (approximately 1 mg, 2-3 mm) tail clips of the banded water snake, Nerodia fasciata, as a means to assess contaminant exposure. The snakes were split into three treatments (n = 8) and were fed three increasing levels (control, medium, and high) of fish contaminated with As, Se, and Sr for 24 months. LA-ICP-MS concentrations of tail clips for all three elements were significantly correlated with the remaining whole tail concentration determined by homogenization, acid digestion, and ICP-MS analysis. Additionally, LA-ICP-MS concentrations for As and Se in the tail clip were similar to the acid digestion solution analysis values for the whole tail, which suggests that these elements are homogeneously distributed. Strontium concentrations were underestimated by LA-ICP-MS when compared to whole tail concentrations. Statistical analysis showed that LA-ICP-MS tail clip concentrations differed significantly according to dietary treatment. Posterior probability error rates from nonparametric discriminant function analysis indicated that LA-ICP-MS analysis of tail clips was useful for predicting exposure to Se with only a 4% probability of misclassification among treatments. Errors associated with misclassification of As were greater (17%) but this was, in part, related to the low concentrations of As in the tail (<1 ppm for the highest treatment). Taken together, the findings from this study suggest that LA-ICP-MS of microdissected tissue shows promise as a nondestructive technique for conservation-minded ecotoxicological studies.

Animals↗