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PubMed1996

Mode of action of Duvernoy's gland extracts from the colubrid Dryadophis bifossatus in the chick biventer cervicis nerve-muscle preparation.

Extracts and secretion of the Duvernoy's gland are venomous, inducing in several cases motor paralysis in experimental animals. The extracts of the Duvernoy's gland from the aglyphous colubrid Dryadophis bifossatus are very toxic, eliciting flaccid paralysis in pigeons, rabbits and Hylae (Brazil and Vellard, 1926). In the present study, the neuromuscular action of the extracts of Duvernoy's gland from this Colubridae was investigated in the chick biventer cervicis nerve-muscle preparation. The muscle was indirectly stimulated with supramaximal pulses, and by addition of acetylcholine or carbachol to the organ bath. Direct muscle stimulation was carried out in curarized preparations. The extracts induced an irreversible neuromuscular blockade and also inhibited irreversibly the contracture of the biventer cervicis produced by either acetylcholine or carbachol. The twitches elicited by direct muscle stimulation were not depressed. These results show that the neurotoxin(s) of the extracts interact with the end-plate cholinergic receptors.

Animals↗
PubMed2005

Snake egg immunoglobulins: biochemical characteristics and adjusted isolation procedure.

Transmission of specific immunoglobulins from mothers to their offspring via the egg is a common phenomenon in egg-laying vertebrates but the occurrence of this phenomenon in reptiles, especially in colubrid snakes, has not been proven until recently. Thus, the essential biochemical characteristics of antibodies deposited in eggs of Elaphe guttata (Colubridae, Serpentes) were studied after isolation of the antibody by precipitation and purification by affinity chromatography and gel filtration, with verification by isoelectric focusing and immunoprecipitation. The immunoglobulin deposited in the eggs of colubrid snakes is a singular, non-truncated IgY antibody in a concentration roughly equal to that in the snake's serum. An efficient method to isolate antibody from snake eggs was developed, based on the PEG precipitation technique of avian yolk immunoglobulins; an unsophisticated protocol for the isolation procedure appropriate for reptile eggs is provided.

Animals↗
PubMed2003

Phylogenetic conservation of a snake venom metalloproteinase epitope recognized by a monoclonal antibody that neutralizes hemorrhagic activity.

Snake venom metalloproteinases (SVMPs) are present in large quantities in venoms of viper snakes and also in some elapids. Jararhagin is a representative of a P-III multidomain hemorrhagic SVMP present in Bothrops jararaca venom. It is comprised of a catalytic, a disintegrin-like and a cysteine-rich domain. Seven anti-jararhagin monoclonal antibodies (MAJar 1-7) were produced, of which six reacted with the disintegrin domain. MAJar 3 recognized an epitope present at the C-terminal part of the disintegrin-like domain, and neutralized jararhagin-induced hemorrhage. In this study, we evaluated the reactivity of these monoclonal antibodies with venoms from 27 species of snakes belonging to different families. MAJar 3 recognized most of the hemorrhagic venoms. By ELISA, MAJar 3 reacted strongly with venoms from Viperidae family and weakly with Colubridae and Elapidae venoms. This recognition pattern was due to bands between 50 and 80 kDa, corresponding to P-III SVMPs. This antibody preferentially neutralized the hemorrhage induced by venoms of Bothrops snakes. This fact suggests that the epitope recognized by MAJar 3 is present in other metalloproteinases throughout snake phylogeny. However, slight structural differences in the epitope may result in insufficient affinity for neutralization of biological activities.

Amino Acid Sequence↗
PubMed2006

Venom of the Brown Treesnake, Boiga irregularis: ontogenetic shifts and taxa-specific toxicity.

The Brown Treesnake (Boiga irregularis), a rear-fanged member of the polyphyletic family Colubridae, is an introduced predator on Guam which has been responsible for numerous human envenomations. Because little is known about this species' venom, we characterized venom proteins from B. irregularis using enzyme assays, one and 2D electrophoresis, Western blot analysis, mass spectrometry, HPLC and toxicity assays. Venom yields and protein content varied significantly with snake size, and large adult specimens averaged over 500 microl venom (19.2 mg, protein content approximately 90%). Only two enzymes, azocaseinolytic metalloprotease and acetylcholinesterase, were detected in venoms, and both activities increased with snake size/age. Western blot analysis demonstrated a 25 kDa CRiSP homolog in venoms from both neonate and adult snakes. 2D electrophoresis showed variation between venoms from neonate and adult snakes, especially with respect to metalloprotease and acetylcholinesterase. Analysis by MALDI-TOF mass spectrometry revealed the presence of numerous proteins with molecular masses of approximately 8.5-11 kDa. Adult B. irregularis venom was quite toxic to domestic chickens (Gallus domesticus; 1.75 microg/g) and lizards (Hemidactylus geckos: 2.5 microg/g and Carlia skinks: 4.5 microg/g), and intoxication was characterized by rapid paralysis of all species and neck droop in chickens. Toxicity of venom from neonates toward geckos was 1.1 microg/g, consistent with the presence of a greater diversity of 8-11 kDa proteins (suspected neurotoxins) in these venoms. All of these values were notably lower than murine LD50 values (neonate: 18 microg/g; adult: 31 microg/g). Like venoms of several front-fanged species, B. irregularis venom showed an ontogenetic shift in enzyme activities and toxicity, and neonate snakes produced more toxic venoms with lower protease and acetylcholinesterase activities. High toxicity toward non-mammalian prey demonstrated the presence of taxa-specific effects (and thus toxins) in B. irregularis venom, likely a characteristic of many colubrid snake venoms. We hypothesize that the lack of significant envenomation effects in humans following most colubrid bites results from this taxa-specific action of colubrid venom components, not from a lack of toxins.

Aging↗
PubMed2002

Identification of beta-type phospholipase A(2) inhibitor in a nonvenomous snake, Elaphe quadrivirgata.

A novel serum protein inhibiting specifically the enzymatic activity of the basic phospholipase A(2) (PLA(2)) from the venom of the Chinese mamushi snake (Agkistrodon blomhoffii siniticus) was purified from a nonvenomous Colubridae snake, Elaphe quadrivirgata. The purified inhibitor was a 150-kDa glycoprotein having a trimeric structure, composed of two homologous 50-kDa subunits. Their amino acid sequences, containing leucine-rich repeats, were typical of the beta-type PLA(2) inhibitor (PLIbeta), previously identified from the serum of A. blomhoffii siniticus. The inhibitor inhibited exclusively group II basic PLA(2)s and did not inhibit other kinds of PLA(2)s. This is the first paper reporting the existence of PLIbeta in a nonvenomous snake. The existence of PLIbeta in the nonvenomous snake reflects that PLIbetas are widely distributed over the snake species and participate commonly in regulating the physiological activities of the unidentified target PLA(2)s.

Amino Acid Sequence↗
PubMed1999

Bites by the colubrid snake Philodryas olfersii: a clinical and epidemiological study of 43 cases.

Less than 10 cases of bites by Philodryas olfersii (Colubridae) have been reported in the literature. In this study, 43 patients admitted to the Instituto Butantan, São Paulo, Brazil, with the diagnosis of P. olfersii bite from 1982 to 1990 were reviewed. The 32 male (74.4%) and 11 female (25.6%) patients presented mainly from November to February (65%). The most common clinical features were local pain (37.2%), swelling (34.9%), erythema (18.6%) and ecchymosis (9.3%). The 20 minute whole blood clotting test was performed in 11 patients and in all of them the blood was coagulable. Most of the accidents occurred during the hottest months and during daylight hours. The most common bite site was the hands. Severe envenoming is not frequent in these accidents.

Adolescent↗
PubMed1997

Detection of calcium-binding proteins in venom and Duvernoy's glands of South American snakes and their secretions.

Calcium-binding proteins (CaBPs) have been described as involved in the stimulus-secretion coupling mechanisms in secretory glands. CaBPs were revealed with 45Ca, after electrophoresis in SDS-PAGE and transference to Zeta probe membranes, in Duvernoy's or venom gland homogenates from three families of South American snakes: Viperidae (Bothrops jararaca and Crotalus durissus terrificus); Elapidae (Micrurus corallinus), and Colubridae (Phylodrias patagoniensis and Oxyrhopus trigeminus). A band with an estimated molecular weight of 12 KDa was found in all glands studied. Bands with 17, 28, and 67 KDa were found in all glands, except in O. trigeminus Duvernoy's gland. A 18 KDa band was found in Viperidae and Elapidae venom glands, and a 88 KDa band was observed only in Viperidae venom gland homogenates. Some of these CaBPs were identified by Western blotting or by immunohistochemistry, as parvalbumin (12 KDa) and calbindin (28 KDa). When the secretion of these glands were analyzed, CaBPs were detected only in B. jararaca venom, with bands of 14, 35, 42, and 72 KDa. The profile of CaBPs was not modified at different phases of the secretory cycle of the glands, as well as after isoproterenol treatment.

Animals↗
PubMed2000

Diet of the colubrid snake, Thamnodynastes strigatus (Günther, 1858) from Paraná State, Brazil, with field notes on anuran predation.

Dissection of 44 specimens of the colubrid snake Thamnodynastes strigatus (Serpentes: Colubridae) provided data on food habits. Prey items include frogs (71.4% of the sample), rodents (14.3%), fishes (10.7%), and lizards (3.6%). The most of the anuran species recorded (Bufo sp., Leptodactylus sp., Physalaemus cuvieri and Scinax fuscovarius) are ground dwellers or call at the water level. Field observations provided information on the feeding behavior.

Animals↗
PubMed2006

Why do most tropical animals reproduce seasonally? Testing hypotheses on an Australian snake.

Most species reproduce seasonally, even in the tropics where activity occurs year-round. Squamate reptiles provide ideal model organisms to clarify the ultimate (adaptive) reasons for the restriction of reproduction to specific times of year. Females of almost all temperate-zone reptile species produce their eggs or offspring in the warmest time of the year, thereby synchronizing embryogenesis with high ambient temperatures. However, although tropical reptiles are freed from this thermal constraint, most do not reproduce year-round. Seasonal reproduction in tropical reptiles might be driven by biotic factors (e.g., peak periods of predation on eggs or hatchlings, or food for hatchlings) or abiotic factors (e.g., seasonal availability of suitably moist incubation conditions). Keelback snakes (Tropidonophis mairii, Colubridae) in tropical Australia reproduce from April to November, but with a major peak in May-June. Our field studies falsify hypotheses that invoke biotic factors as explanations for this pattern: the timing of nesting does not minimize predation on eggs, nor maximize food availability or survival rates for hatchlings. Instead, our data implicate abiotic factors: female keelbacks nest most intensely soon after the cessation of monsoonal rains when soils are moist enough to sustain optimal embryogenesis (wetter nests produce larger hatchlings, that are more likely to survive) but are unlikely to become waterlogged (which is lethal to eggs). Thus, abiotic factors may favor seasonal reproduction in tropical as well as temperate-zone animals.

Analysis of Variance↗
PubMed2006

Characterization of a novel metalloproteinase in Duvernoy's gland of Rhabdophis tigrinus tigrinus.

During the characterization of hemorrhagic factor in venom of Rhabdophis tigrinus tigrinus, so-called Yamakagashi in Japan, one of the Colubridae family, a novel metalloproteinase with molecular weight of 38 kDa in the Duvernoy's gland of Yamakagashi was identified by gelatin zymography and by monitoring its proteolytic activity using a fluorescence peptide substrate, MOCAc-PLGLA2pr(Dnp)AR-NH2, which was developed for measuring the well-known matrix metalloproteinase (MMP) activity. After purification by gel filtration HPLC and/or column switch HPLC system consisting of an affinity column, which was immobilized with a synthetic BS-10 peptide (MQKPRCGVPD) originating from propeptide domain of MMP-7 and a reversed-phase column, the N-terminal amino acid sequence of the 38 kDa metalloproteinase was identified as FNTFPGDLK which shared a high homology to Xenopus MMP-9. The 38 kDa metalloproteinase required Zn2+ and Ca2+ ions for its proteolytic activity. In addition, the proteolytic activity was almost completely inhibited by BS-10, a MMP inhibitor, but not by the serine proteinase inhibitors, cysteine proteinase inhibitors and aspartic proteinase inhibitors. Together these results demonstrated that the 38 kDa proteinase is a novel snake verom metalloproteinase (SVMP) containing HExGHxxGxxH motif which possesses high affinity to the BS-10 peptide, into its molecule, and the enzymatic properties are closed to that of MMPs. Based on the results obtained in the present study, we concluded that the 38 kDa metalloproteinase is a novel metalloproteinase whose activity may be regulated by the cysteine switch mechanism, and could be classified as one of the matrix metalloproteinases rather than snake venom metalloproteinases.

Animals↗
PubMed1994

[Use of a body condition index (BCI) for the study of the reproduction in snakes].

A body condition index (BCI = actual body mass/optimal theoritical body mass of the studied animal) was estimated in females of 3 species of snake. From dissections of 88 Vipera aspis and 18 Coluber viridiflavus, strong relationships between body reserves (such as mass of fat bodies and liver) and BCI were found. Furthermore, BCI explained most of the variance in the mass of the fat bodies and in the mass of the liver; however BCI and body length together increased the percentage of variation explained. Thus, a satisfactory estimation of body reserves in relation to body length is possible in living snakes. We have studied reproductive parameters (clutch size and litter size) in Vipera aspis (Viperidae) and Elaphe longissima (Colubridae) during a 3 year period. In the two species positive relationships between maternal body length and number of offspring were found. At the beginning of vitellogenesis, litter size is related to the BCI level in V. aspis but not in E. longissima. In both species, BCI must exceed a threshold value for reproduction to take place. However this threshold value is much lower in E. longissima (0.55) than in V. aspis (0.70). This indicates that in the latter species, large body reserves are necessary for the induction of vitellogenesis. In E. longissima, maternal body length is an important determinant of reproductive success, body reserves playing a minor role. In contrast, in female V. aspis the reproductive success is related to BCI and to a lesser extent to body length.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

[Poisonous snakes of Europe].

Characteristics of poisonous snakes and their toxines are described. The appearance and biology of all European poisonous snakes, eight vipers (family Viperidae) and one opisthoglyph colubride snake (family Colubridae) are given in detail.

Agkistrodon↗
PubMed2005

Ultrastructural evidence for secretion from the epithelium of ampulla ductus deferentis of the fan-throated lizard Sitana ponticeriana Cuvier.

Among reptiles, an ampulla ductus deferentis has been reported only in Squamata. Fairly detailed studies are available only for two species, the lizard Calotes versicolor (Fam: Agamidae) and the snake Seminatrix pygaea (Fam: Colubridae). The light microscopic study on C. versicolor revealed the ampulla to be a prominent organ, whereas the light and transmission electron microscopic study in S. pygaea revealed it to be discernable only in histological preparations. Further, the epithelium of the ductal portion of vas deferens as well as the ampulla of C. versicolor appears to contribute to the seminal plasma and can also phagocytose dead sperm, whereas in S. pygaea neither of these roles has been established. Thus, we hypothesize that there may be variations in the anatomy, histology, and the role of the vas deferens in general, and the ampulla in particular, of the squamate reptiles. In this study, the ductus deferens of the small fan-throated lizard Sitana ponticeriana (Fam: Agamidae) was subjected to light and transmission electron microscopic analysis. In this lizard the ampulla is more prominent than in C. versicolor. The epithelium of the ductal portion of vas deferens consists of principal cells (with features reflecting roles in endocytosis and phagocytosis of dead sperm), dark cells (which are absent in the epithelium of the ductal portion of vas deferens of snakes), and basal cells. The ampulla of S. ponticeriana is differentiated into storage and glandular portions. The epithelium of the storage portion is like that in the ductal portion of the vas deferens, whereas that of the glandular portion, consisting of dark and light principal cells and foamy cells, is tall and forms into smooth villous folds. All three cell types show evidence for a role in secretion, in all likelihood different from each other, for release into the lumen to contribute to seminal plasma. These cells do not provide evidence of a role in phagocytosis of dead sperm. It appears that within the Squamata, the ductal ampulla differs in structure as well as function. We suggest that the ductal ampulla of agamid lizards is a composite gland of the ampulla ductus deferentis and seminal vesicles of mammals.

Animals↗
PubMed1976

Satellite DNA and evolution of sex chromosomes.

The satellite DNA (SATELLITE III) which is mainly represented in the female of Elaphe radiata (Ophidia, Colubridae) has been isolated and its buoyant density has been determined (delta equals 1.700 g cm-3). In situ hybridisation of radioactive complementary RNA of this satellite DNA with the chromosomes of different species has revealed that it is mainly concentrated on the W sex chromosome and its sequences are conserved throughout the sub-order Ophidia. From hybridisation studies these sequences are absent from the primitive family Boidae which represents a primitive state of differentiation of sex chromosomes. Chromosome analysis and C-banding have also revealed the absence of heteromorphism and of an entirely heterochromatic chromosome in the species belonging to the primitive family and their presence in the species of highly evolved families. It is suggested that the origin of satellite DNA (satellite III) in the W chromosome is the first step in differentiation of W from the Z in snakes by generating asynchrony in the DNA replication pattern of Z and W chromosomes and thus conceivably reducing the frequency of crossing-over between them which is the prerequisite of differentiation of sex chromosomes. Presence of similar sex chromosome associated satellite DNA in domestic chicken suggests its existence in a wider range of vertebrates than just the snakes.

Animals↗
PubMed1991

Conservative and variable regions of homologous snake phospholipases A2 sequences: prediction of the taxon-specific peptides structure.

Homologous amino acid sequences of phospholipases A2 (PLA2) of snakes belonging to the families Elapidae, Viperidae, and Colubridae were considered in order to study the conservative and variable regions location. The PLA2 sequences were divided into two groups (taxons) according to the phylogenetic tree reconstructed from the pair similarity matrix. Results of the intergroup comparison were plotted to facilitate the identification of significant conservative and variable regions. It was shown that the results of the comparison between two phylogenetic groups of snake PLA2 did not much depend on the number of each group representatives and did not markedly change if one of the groups was represented by the single sequence. The knowledge of the number and location of conservative and variable regions and their dependence on the phylogenetic relations between compared taxa may be used to predict a synthetic peptide structure to obtain specific antibodies against PLA2 of one of these taxons. Such prediction is possible if there is a specific region conservative for one taxon but variable for two of them.

Amino Acid Sequence↗
PubMedDate not supplied

Snake envenomation. Incidence, clinical presentation and management.

Snake envenomation is a major cause of death and disability in the developing countries, particularly India and Southeast Asia. Species variation in venom components, yield, and lethality leads to quite different clinical presentations and mortality. Venomous snakes are divided into 5 families. Bites of the Viperidae, Crotalidae and Colubridae usually cause primarily local effects and bleeding; the Elapidae most commonly cause neurological symptoms, particularly paralysis; while the Hydrophidae cause paralysis and myolysis. Venoms are complex mixtures of enzymes, peptides and metalloproteins. 26 enzymes have been identified, and 10 of those are found in most venoms. Components have been identified that act as procoagulants, anticoagulants, hyaluronidases, RNases, DNases, postsynaptic toxins and presynaptic toxins. Other peptides induce capillary leak syndrome, haemolysis and shock. The clinical results of envenomation vary widely, and there may be no envenomation with a bite. Syndromes reported include oedema, haemolysis, shock, bleeding, pituitary failure, renal failure, myonecrosis, and combinations of the above. First aid measures that have been proposed include tourniquets, constricting bands, tight crepe bandages, incision and suction, cryotherapy, and high voltage electric shock. None of these has been shown to be effective except usage of a crepe bandage for Australian elapid bite. Tourniquets or cryotherapy, if used for extended periods may lead to gangrene. The most important first aid measure is rapid transport to comprehensive medical care. There is some controversy about medical treatment in the United States, but less in other countries. Supportive measures routinely required include intravenous fluids, tetanus prophylaxis and antibiotics. Anticholinergics may be useful in elapid bite. Intubation and ventilation may be necessary. Unproven surgical approaches include excision of envenomated tissues and fasciotomy. The former is disfiguring, the latter should be reserved for those patients with demonstrated increased intracompartmental pressure. More than 100 antivenins are produced by about 36 laboratories worldwide. The products are effective, but carry a high risk of serum sickness and a lesser risk of anaphylaxis. A more effective and less reactive product is under development.

Africa↗
PubMed2005

Complete mitochondrial DNA sequences of six snakes: phylogenetic relationships and molecular evolution of genomic features.

Complete mitochondrial DNA (mtDNA) sequences were determined for representative species from six snake families: the acrochordid little file snake, the bold boa constrictor, the cylindrophiid red pipe snake, the viperid himehabu, the pythonid ball python, and the xenopeltid sunbeam snake. Thirteen protein-coding genes, 22 tRNA genes, 2 rRNA genes, and 2 control regions were identified in these mtDNAs. Duplication of the control region and translocation of the tRNALeu gene were two notable features of the snake mtDNAs. The duplicate control regions had nearly identical nucleotide sequences within species but they were divergent among species, suggesting concerted sequence evolution of the two control regions. In addition, the duplicate control regions appear to have facilitated an interchange of some flanking tRNA genes in the viperid lineage. Phylogenetic analyses were conducted using a large number of sites (9570 sites in total) derived from the complete mtDNA sequences. Our data strongly suggested a new phylogenetic relationship among the major families of snakes: ((((Viperidae, Colubridae), Acrochordidae), (((Pythonidae, Xenopeltidae), Cylindrophiidae), Boidae)), Leptotyphlopidae). This conclusion was distinct from a widely accepted view based on morphological characters in denying the sister-group relationship of boids and pythonids, as well as the basal divergence of nonmacrostomatan cylindrophiids. These results imply the significance to reconstruct the snake phylogeny with ample molecular data, such as those from complete mtDNA sequences.

Animals↗
PubMed1990

Isolation of a galactose-binding lectin from the venom of the snake Bothrops godmani (Godmann's pit viper).

A galactose-binding lectin, isolated from the venom of B. godmani by affinity chromatography, is an acidic protein (pI 4.9) with a subunit mol. wt of about 14,000, occurring mostly as a disulfide-linked dimer of 28,000. A small proportion of lectin appears as a monomer and as a tetramer. The lectin agglutinates erythrocytes from mice, rabbit, cow and human (all ABO types, either Rh positive or negative), but does not agglutinate horse, sheep, goat and snake (Oxybelis aeneus, Colubridae) erythrocytes. The agglutinating activity is inhibited by 1 mM EDTA. The lectin is devoid of lethal, hemorrhagic, myotoxic, proteolytic and phospholipase A2 activities. It is not mitogenic for human peripheral blood mononuclear cells. The only effect observed was a moderate induction of edema in the footpad of mice, with a minimal edema-forming dose of 22 micrograms. This effect developed rapidly, and was significantly inhibited by i.p. administration of cyproheptadine, a histamine and serotonin antagonist, before injection of the lectin. Despite the edema-forming activity observed, the low concentration of lectin in crude venom, together with its relatively low potency, suggest that this lectin is not a key component in the development of edema following envenomations by B. godmani.

Animals↗