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Chlorinated hydrocarbon concentrations in plasma of the Lake Erie water snake (Nerodia sipedon insularum) and northern water snake (Nerodia sipedon sipedon) from the Great Lakes basin in 1998.

From the Great Lakes basin, concentrations of 59 congener-specific polychlorinated biphenyls (PCBs) and 14 organochlorine pesticides were measured in blood plasma of northern water snake (Nerodia sipedon sipedon) and Lake Erie water snake (Nerodia sipedon insularum), which is endangered in Canada. In 1998, four male adult Lake Erie water snakes were sampled from Pelee Island, western Lake Erie; four male northern water snakes were sampled at Little Lake, about 20 km north of Parry Sound in central Ontario; and four adult gravid female northern water snakes were sampled from Garden Island, eastern Lake Ontario. The blood plasma was pooled by site for a total of three samples analyzed. The Pelee Island sample from male Lake Erie water snakes contained less than half the lipid concentration (0.349%) than samples from the other sites, but it was the most contaminated with PCBs, even on a wet weight basis. Summed concentration of individual PCBs in the Pelee Island sample was 167 ng/g (wet weight), which was 14-fold higher than the next most contaminated sample, which was from Little Lake. The plasma sample from Little Lake contained 12 ng/g (WW) and was four times more contaminated with PCBs than the sample from female snakes from Garden Island, Lake Ontario. Organochlorine pesticide concentrations in plasma were relatively similar among sites. None of the pesticides was found above trace concentrations (0.1-0.9 ng/g) except pp'-DDE, which occurred at 2-5 ng/g among sites. PCB congener patterns in the Lake Erie water snakes were compared to PCB patterns in plasma of common snapping turtle (Chelydra serpentina serpentina) from Lake Ontario, herring gull eggs (Larus argentatus) from western Lake Erie, and mudpuppy eggs (Necturus maculosus) from the Detroit River. The PCB patterns in water snake and herring gull sample were most similar, followed by the pattern in snapping turtle plasma. The presence of more lower-chlorinated chlorobiphenyls in the mudpuppy eggs relative to the other species made this sample distinct from the water snake, gull, and turtle.

Animals↗

Thermal biology characteristics of the Caspian whipsnake (Dolichophis caspius, Gmelin, 1789) under natural and laboratory conditions.

Reptiles rely on behavioural thermoregulation to maintain optimal body temperature (Tb), essential for many physiological functions. However, thermal biology remains poorly studied in many snake species, including the Caspian whipsnake (Dolichophis caspius, Gmelin, 1789). This study examined the thermal biology of the Caspian whipsnake under natural and laboratory conditions. In the field, we investigated the influence of sex, age category, season, time of day, air temperature, detection status, feeding status, and shedding phase on Tb. Under controlled conditions, we assessed preferred Tb variation and its relationship to active thermoregulation. In the field, Tb was significantly affected by sex, season, time of day, air temperature, feeding status, and shedding phase, but not age category, body size or detection status. Males had lower Tb than females and snakes with food in their stomachs exhibited higher Tb. Tb increased from March to June and from morning to afternoon, and air temperature had a positive effect on Tb. In the laboratory, mean Tb (24.5 ± 4.9 °C) was lower than in the field (26.9 ± 5.2 °C) and was significantly associated with body posture and terrarium part, indicating active behavioural thermoregulation. In the laboratory, males had lower Tb than females, and juveniles had lower Tb than adults. The lower thermal preference suggests a cooler thermal niche than many colubrids, potentially increasing their susceptibility to climate warming. However, the broad Tb variation and behavioural thermoregulation indicate some flexibility in responding to environmental changes. These findings highlight the role of behavioural thermoregulation in snake thermal responses to environmental variation.

Behavioural thermoregulation↗

Ca2+ and Ba2+ effects on basolateral tetraethylammonium transport in isolated snake renal proximal tubules.

Previous work on snake renal tubules suggested that basolateral transport of tetraethylammonium (TEA) is symmetrical. To examine regulation of this transport step more closely, we determined the effects of (1) reductions in the extracellular Ca2+ concentration and basolateral Ca2+ entry, and (2) the presence of extracellular Ba2+ on TEA uptake and efflux across the basolateral membrane of isolated snake renal proximal tubules. Removal of extracellular Ca2+ reduced initial TEA uptake and enhanced TEA efflux. Blocking Ca2+ entry also reduced initial TEA uptake. Extracellular Ba2+ depolarized the basolateral membrane and reduced both TEA uptake and efflux. Inhibition of basolateral TEA uptake with a reduced membrane potential supports previous data indicating that uptake involves potential-driven facilitated diffusion. Inhibition of basolateral TEA efflux by Ba2+ even with a reduced membrane potential not only supports previously obtained data indicating that efflux is not influenced by the potential difference and that basolateral TEA transport is asymmetrical, but also suggests that TEA uptake and efflux may occur by separate pathways.

Animals↗

Ultrastructure of Hepatozoon boigae (Mackerras, 1961) nov. comb. from brown tree snakes, Boiga irregularis, from northern Australia.

Intraerythrocytic bodies identified as haemogregarine gamonts were found in 29% of 97 brown tree snakes (Boiga irregularis) examined during a haematological survey of reptiles in Australasia during 1994-1998. The morphological characteristics of the parasites were consistent with those of Haemogregarina boigae Mackerras, 1961, although the gamonts were slightly larger and lacked red caps but contained distinctive polar grey capsules. Gamonts did not distend host cells but laterally displaced their nuclei. They were contained within parasitophorous vacuoles and possessed typical apicomplexan organelles, including a conoid, polar rings, rhoptries and micronemes. Schizonts producing up to 30 merozoites were detected in endothelial cells of the lungs of 11 snakes. The absence of erythrocytic schizogony suggests the parasites belong to the genus Hepatozoon. Electron microscopy also revealed the presence of curious encapsulated organisms in degenerating erythrocytes. These stages did not possess apical complex organelles and were surrounded by thick walls containing circumferential junctions and interposed strips reminiscent of oocyst sutures.

Animals↗

Patterns of avian nest predators and a brood parasite among restored riparian habitats in agricultural watersheds.

In fragmented edge-dominated landscapes, nest predation and brood parasitism may reduce avian reproductive success and, ultimately, populations of some passerine species. In the fragmented agroecosystem of northwest Mississippi, placement of drop-pipe structures has been used as a restoration technique for abating gully erosion along stream banks. These actions have formed small herbaceous and woody habitat extensions into former agricultural lands. We quantified species relative abundances, species richness, and evenness of avian nest predators and a brood parasite within four categories of constructed habitat resulting from drop-pipe installation. Differences in the abundance of two nest predators, cotton mouse (Peromyscus gossypinus) and blue jay (Cyanocitta cristata), were observed among constructed habitats. However, relative abundances of other predators such as common grackle (Quiscalus quiscula), American crow (Corvus brachyrhynchos), and hispid cotton rat (Sigmodon hispidus), and the obligate brood parasite brown-headed cowbird (Molothrus ater) did not differ among four habitat categories. Although species richness, abundance, and evenness of potential nest predators were generally similar among the constructed habitats, predator species composition varied, suggesting that these habitats supported different predator communities. This difference is important because as each predator species is added to or deleted from the community, variation may occur in the framework of prey search methods, predator strategies, and potentially overall predation pressure. We suggest that land managers using drop-pipes as part of stream restoration projects allow for the development of the constructed habitat with the largest area and greatest vegetative structure.

Agriculture↗

Seven new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from colubrid snakes of Guatemala and a discussion of what to call ellipsoid tetrasporocystic, dizoic coccidia of reptiles.

During a survey of Guatemalan herpetofauna in the summers of 1998-2000, 29 presumed new species of Eimeria Schneider, 1875 were found, seven of which have a distinct elongate-ellipsoidal shape (L/W ratio >or= 1.7) and are described herein. Six of the seven new species are similar in oöcyst length, width and L/W ratio and sporocyst length, width and L/W ratio, lack a micropyle, oöcyst residuum, Stieda body, sub-- and parastieda bodies, have a polar granule and sporocyst residuum, and their sporocysts appear to have dehiscence sutures. The seventh is slightly smaller and has sporocysts with a Stieda body. The new species are: E. coniophanes n. sp - whose sporulated oöcysts from Coniophanes fissidens are 29.2x14.9 (27-31x13-16) microm, with sporocysts 10.0 x 7.8 microm; E. coniophis n. sp. -from Conophis lineatus are 32.0x16.5 (30-34x14-18) microm, with sporocysts 10.2 x 8.9microm; E. dryomarchoni n. sp. - from Drymarchon corais are 32.2x17.7 (31-34x17-19) microm, with sporocysts 10.7 x 8.6 microm; E. leptophis n. sp. - from Leptophis mexicanus are 29.5x17.0 (28-31x16-18) microm, with sporocysts 10.0 x 9.1 microm; E. oxybelis n. sp. - from Oxybelis aeneus are 31.8x16.5 (29-33x15-18) microm, with sporocysts 10.3 x 8.8 microm; and E. scaphiodontophis n. sp. - from Scaphiodontophis annulatus are 30.0x15.3 (28-33x14-16) microm, with sporocysts 9.9 x 7.9 microm. Sporulated oöcysts of E. siboni n. sp. from Sibon nebulata are 24.3x14.2 (21-27x13-16) microm, with sporocysts 10.0 x 7.1 microm and with a Stieda body. We conclude that until all aspects of each life-cycle are known, it is prudent at this time to name all tetrasporocystic dizoic coccidia from snakes as members of Eimeria rather than place some of them in Choleoeimeria Paperna & Landsberg, 1989.

Animals↗

Factors governing the appearance of small-mode miniature endplate currents at the snake neuromuscular junction.

At the snake neuromuscular junction, following a short period of high frequency motor nerve stimulation, a population of 'small-mode' miniature endplate currents (MEPCs) is seen. These small-mode MEPCs are believed to be due to the release of acetylcholine from incompletely refilled recycling synaptic vesicles [Searl et al., Neuroscience, 35 (1991) 145-156]. This study determines the role of the trans-vesicular membrane proton gradient in the generation of small-mode MEPCs. Preserving the trans-vesicular membrane proton gradient during synaptic vesicle exocytosis by exposure of snake nerve/muscle preparations to an extracellular pH approximately equal to the intravesicular pH, leads to an augmentation of the amplitude of the stimulation-induced small-mode MEPCs. Conversely, pretreatment of snake neuromuscular preparations with 10 mM ammonium ions, to buffer intravesicular protons and hence dissipate the trans-vesicular membrane proton gradient, leads to an inhibition of the stimulation-induced appearance of small-mode MEPCs. The data are consistent with the theory that the reduced acetylcholine content of recycled synaptic vesicles in the snake is a consequence of an incomplete restoration of the trans-vesicular membrane proton gradient following synaptic vesicle exocytosis. Thus, in recycling synaptic vesicles the limiting step in the refilling process appears to be generation of the trans-membrane proton gradient and not the transport of acetylcholine into the vesicle.

Ammonium Chloride↗

Feeding behavior and an oropharyngeal component of satiety in a two-headed snake.

We studied the feeding behavior of a dicephalous Black Rat Snake (Elaphe o. obsoleta) whose heads attack and ingest prey independently. Both heads were given satiation feedings with mouse prey at either 5-day intervals (1989, n = 16 feedings) or 9-11 day intervals (1990, n = 4 feedings). In 1989, a meal pattern was manifested as a negative correlation between total (left + right) meal size (g) and total size of the last meal. Meal sizes of the heads were consistently different (L > R) despite their apparently identical genotypes. We confirmed tongue-flicking rate and swallowing time as indices of satiety/hunger in each head. In order to test whether oropharyngeal stimulation is involved in satiation, we analyzed three measures: willingness to feed, and intrameal changes in swallowing, and tongue-flicking rates. Data for these tests were taken from experimental feedings in which one head was fed to satiation (first head) before the other head (second head) was fed. The second head generally did eat (8 of 10 experimental feedings) after the first head had become satiated. In addition, while the time to swallow a mouse generally increased as the snake neared satiety, swallowing time decreased significantly (p = 0.04) from the first head's last mouse to the second head's first mouse. Finally, in two of three feedings, the first head's tongue-flicking rate decreased before the second head's. These results, which take advantage of this unique animal, indicate that oropharyngeal stimulation is involved in meal termination.

Animals↗

Snake odor alters behavior, but not pain sensitivity in mice.

It is known that predator cues (both mammalian odor or avian vocalization) elicit marked analgesia in rodents. The present experiment used olfactory cues produced by an opportunistic rodent predator snake species (100 cc of sawdust scented by Elaphe quatuorlineata). Upon exposure to snake odor (for 30 s, 20 min, or 40 min), adult mice of both the CD-1 and DBA2 strains were assessed for tail-flick or hot-plate analgesia at different times after exposure (from 0 to 40 or 45 min, respectively). In both strains, snake odor exposure induced significant alteration in the frequencies of sniffing, self-grooming, and digging, while it inhibited habituation of locomotor activity in DBA/2 mice. No analgesia emerged with both tests as a consequence of exposure to snake odor. Results suggest that although endogenous analgesia has been demonstrated by other authors to be elicited in response to cues emanating from common and widely distributed mouse predators (such as carnivores or owls), predators such as reptiles, which under natural conditions exert a limited predatory pressure on the house mouse gene pool, may only induce fear-associated behavioral responses but cannot provide ethologically relevant stimuli triggering mouse analgesia.

Animals↗

Effects of monoamine oxidase inhibitors and dopamine agonists on the behavior of mammal- and frog-eating snakes.

Skin mucus of the frog, Xenopus laevis, induces climbing and attenuates tongue flicking in Nerodia sipedon; these effects are induced alone and are potentiated by L-deprenyl, a monoamine oxidase type B inhibitor (MAO-Bi), but not by clorgyline, an MAO type A inhibitor (MAO-Ai). Both MAO-A and MAO-B metabolize dopamine, with MAO-B having the higher affinity; MAO-A selectively metabolizes serotonin and norepinephrine and MAO-B is selective for phenylethylamine. It was hypothesized that clorgyline and L-deprenyl would differentially modulate tongue flicking and climbing in frog-eating (Nerodia erythrogaster) and mammaphagous (Elaphe o. obsoleta) snakes, based on physiological differences between the species. L-Deprenyl caused a decrease in tongue flicking and climbing by Elaphe and an increase in climbing by Nerodia, whereas clorgyline did not alter tongue flicking, climbing, or locomotor activity in either species. To further assess the role of dopamine, hybrid black/gray rat snakes, E. o. spiloides, were administered the D1 and D2 dopamine receptor agonists SKF 77434 (SKF 38393, N-allyl) and quinpirole, respectively. SKF 77434 and quinpirole attenuated climbing, but only SKF 77434 attenuated tongue flicking in Experiment 3; neither drug affected locomotor activity. Results suggest that dopaminergic stimulation by MAO-Bi and dopamine agonists modulates tongue flicking and climbing behaviors in snakes, and that the contrasting climbing reactions induced by MAO-Bi between Elaphe and Nerodia may be linked to quantitative differences in endogenous catecholamine levels and/or to the numbers and sensitivity of receptors.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗

Isolation and characterization of five fibrin(ogen)olytic enzymes from the venom of Philodryas olfersii (green snake).

Five distinct fibrin(ogen)olytic proteinases PofibC1, C2, C3, H and S were isolated by gel filtration and ion-exchange chromatographies. PofibC1, C2, C3 and H are metalloproteinases inhibited by ethylenediamine tetracetic acid (EDTA) or 1,10-phenanthroline. Only PofibH had hemorrhagic activity. PofibS is a serine proteinase, inhibited by phenylmethylsulfonyl fluoride (PMSF) or Torresea cearensis trypsin inhibitor (TCTI). All five enzymes were inhibited by dithiothreitol (DTT) or dithioerythritol (DTE). PofibC1 and C2 presented the same mol. wt of 47,000 and are acidic proteins of pI 6.2 PofibC3 is a basic proteinase of pI 8.5 and mol. wt 45,000. The hemorrhagic proteinase PofibH had a mol. wt of 58,000 and pI of 4.6 and PofibS had a mol. wt of 36,000 and pI of 4.5. The five proteinases degraded fibrin and fibrinogen. PofibC1, C2, C3 and H degraded preferentially A alpha-chains while PofibS cleaved concomitantly A alpha and B beta-chains of fibrinogen. None of these enzymes cleaved the gamma-chain of fibrinogen. When correlated with the thrombin delay time, the most active was PofibS, while PofibH and PofibC1 showed almost no activity. The proteinases also differed in the peptide cleavage of B-chain of insulin. Philodryas olfersii venom promoted in vivo a loss of the circulant plasma fibrinogen, as was observed in experiments with rats.

Amino Acid Sequence↗

Properties of Duvernoy's secretions from opisthoglyphous and aglyphous colubrid snakes.

Relatively little attention has been given to the biological properties of Duvernoy's secretions produced by opisthoglyphous and some aglyphous colubrid snakes. A review is presented of literature pertaining to these secretions. Most detailed analyses of Duvernoy's secretions and their biological properties have been performed since the late 1970s. The dispholidines, Dispholidus typus and Thelotornis sp., and the natricines, Rhabdophis tigrinus and R. subminiata, have received the most attention due to the high toxicity of their secretions and their medical importance. These species produce secretions with variably strong prothrombin-activating activity, defibrinating activity, and hemorrhagic potential. Boigines, and natricines other than Rhabdophis, produce secretions of low to moderate toxicity and are variably hemorrhagic and proteolytic. Xenodontines and homalopsines similarly show hemorrhagic potential with low to moderate toxicity. Neurotoxic activity has been reported only from secretions of the boigines, Boiga blandingi and B. irregularis and the xenodontine, Heterodon platyrhinos. These species produce secretions containing postsynaptically acting components. Analyses of some of these secretions have shown that enzymes common to many ophidian venoms such as phospholipases A and L-amino acid oxidase are uncommon in the colubrid secretions studied. This may be due to few studies assaying for multiple enzyme activities and/or the unavailability of many secretion samples for study. Methods of secretion extraction, storage, and assay are discussed. Projected future research and the adaptive implications of Duvernoy's secretions are considered.

Animals↗

The effects of Duvernoy's gland secretion from the xenodontine colubrid Philodryas olfersii on striated muscle and the neuromuscular junction: partial characterization of a neuromuscular fraction.

The effect of Philodryas olfersii Duvernoy's secretion was studied in vivo in mice and chicks as well as in the mouse phrenic nerve-diaphragm and the chick biventer cervicis preparations. The whole secretion (20-40 micrograms/ml) increased the creatine kinase (CK) levels in mice but had no effect on the mouse phrenic nerve-diaphragm preparation. In the chick, the secretion caused head drop and paresia as well as irreversible blockade of the twitch-tension evoked by indirect stimulation in the chick biventer cervicis preparation (50% paralysis in 34.5 +/- 2.7 min, n = 4). The secretion also caused muscle contracture (30% of the maximal twitch-tension generated) after a latency of nearly 9 min. Following fractionation on a Superose 12 FPLC column, the neuromuscular activity was recovered in the high mol. wt fraction (Peak I). At a concentration of 10 micrograms/ml in the chick biventer cervicis preparation, Peak I caused 50% paralysis within 18.5 +/- 3.0 min (n = 4), and evoked a strong contracture (70% of the maximal twitch-tension generated). The contractile responses of the chick preparation to ACh and KCL were partially blocked (90%) by the whole secretion and totally blocked by Peak I. CK release was increased by the whole secretion but not by Peak I. The whole secretion also produced various degrees of muscle cell lysis and extensive widening of the intercellular spaces. The latter showed a loosely arranged membranous network. In general, Peak I caused only minor morphological alterations compared with the whole secretion, although these were still significantly different from those observed in the control preparations. The changes principally involved hypercontraction of the muscle fibers. Based on the above results, we conclude that Peak I contains the factor(s) responsible for the in vitro effects on neuromuscular transmission, whereas the direct myotoxic effect is apparently caused by at least one other component of the Duvernoy's secretion.

Animals↗

Healing rituals and sacred serpents.

Votive tablets found during the excavation of shrines of the Graeco-Roman god of medicine (Asklepios or Aesculapius) associate the healing of superficial lesions with contact with the oral cavity of non-poisonous serpents. We suggest that this may have been the empirical exploitation of the healing properties of salivary growth factors. By immunohistochemistry and immunoblotting we demonstrate the expression of the epidermal growth factor and its receptor in the oral, upper digestive, and salivary epithelia of Elaphe quatuorlineata, a species probably used in healing rituals.

Animals↗

A potent antihemorrhagin in the serum of the non-poisonous water snake Natrix tessellata: isolation, characterization and mechanism of neutralization.

The main natural antihemorrhagic factor (NtAH), which inhibits the hemorrhagic activity of Bothrops asper snake venom, was isolated from the serum of the non-poisonous water snake Natrix tessellata by ammonium sulfate precipitation at 35-55%, Sephadex G-75 gel filtration, ion exchange chromatography on DEAE-Sepharose and CM-Sepharose and hydrophobic Phenyl-Sepharose chromatography. The purified protein showed one band with an isoelectric point of 4.5 and a molecular mass of about 880 kDa. The antihemorrhagic activity was stable between pH 5.5-11.7 and up to 50 degrees C, but lost activity after 20 min at 60 degrees C. It did not form a precipitin line with the main hemorrhagin of Bothrops asper snake venom (BaH1), nor with the whole venom, which suggests that the antihemorrhagic factor is not an immunoglobulin. The mechanism of neutralization by the isolated antihemorrhagic factor NtAH did not include digestion of the hemorrhagic toxin BaH1. Chromatography of NtAH with active 125I-labeled BaH1 toxin as well as ELISA experiments demonstrated that the mechanism of neutralization involves formation of an inactive soluble complex between the natural NtAH of the non-poisonous water snake and the main hemorrhagin of Bothrops asper venom.

Animals↗

Influence of physical stress, distress cues, and predator kairomones on the foraging behavior of Ozark zigzag salamanders, Plethodon angusticlavius.

An animal's response to environmentally induced stress can have important fitness consequences. For example, an animal may reduce activity in response to stress related to increased predation risk, but this response carries the potential cost of missed foraging or mating opportunities. We used laboratory bioassays to determine whether Ozark zigzag salamanders, Plethodon angusticlavius, decreased their feeding behavior following increased levels of threat-related stress. Stress level was manipulated in three experiments where salamanders were exposed to direct or indirect indicators of stress. The direct cue was a physical attack. The indirect cues were chemical cues from distressed conspecifics and chemical cues ("kairomones") from predatory ringneck snakes, Diadophis punctatus. For all three cues, stressed salamanders showed lower foraging success than unstressed salamanders. The physical attack did not result in a more intense response than the other treatments, possibly because the attack occurred away from the foraging area. The intensity of the response to stress may depend more on the nature of the threat (identity and imminence) than the level of physical disturbance.

Adaptation, Physiological↗

Experimentally reducing clutch size reveals a fixed upper limit to egg size in snakes, evidence from the king ratsnake, Elaphe carinata.

Snakes are free of the pelvic girdle's constraint on maximum offspring size, and therefore present an opportunity to investigate the upper limit to offspring size without the limit imposed by the pelvic girdle dimension. We used the king ratsnake (Elaphe carinata) as a model animal to examine whether follicle ablation may result in enlargement of egg size in snakes and, if so, whether there is a fixed upper limit to egg size. Females with small sized yolking follicles were assigned to three manipulated, one sham-manipulated and one control treatments in mid-May, and two, four or six yolking follicles in the manipulated females were then ablated. Females undergoing follicle ablation produced fewer, but larger as well as more elongated, eggs than control females primarily by increasing egg length. This finding suggests that follicle ablation may result in enlargement of egg size in E. carinata. Mean values for egg width remained almost unchanged across the five treatments, suggesting that egg width is more likely to be shaped by the morphological feature of the oviduct. Clutch mass dropped dramatically in four- and six-follicle ablated females. The function describing the relationship between size and number of eggs reveals that egg size increases with decreasing clutch size at an ever-decreasing rate, with the tangent slope of the function for the six-follicle ablation treatment being -0.04. According to the function describing instantaneous variation in tangent slope, the maximum value of tangent slope should converge towards zero. This result provides evidence that there is a fixed upper limit to egg size in E. carinata.

Animals↗