Crotoxin: Novel activities for a classic beta-neurotoxin

Crotoxin, the main toxin of South American rattlesnake (Crotalus durissus terrificus) venom, was the first snake venom protein to be purified and crystallized. Crotoxin is a heterodimeric beta-neurotoxin that consists of a weakly toxic basic phospholipase A(2) and a nonenzymatic, non-toxic acidic component (crotapotin). The classic biological activities normally attributed to crotoxin include neurotoxicity, myotoxicity, nephrotoxicity and cardiotoxicity. However, numerous studies in recent years have shown that crotoxin also has immunomodulatory, anti-inflammatory, anti-microbial, anti-tumor and analgesic actions. In this review, we describe the historical background to the discovery of crotoxin and its main toxic activities and then discuss recent structure-function studies and investigations that have led to the identification of novel pharmacological activities for the toxin. (C) 2010 Elsevier Ltd. All rights reserved.

Identification of continuous interaction sites in PLA(2)-based protein complexes by peptide arrays

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparison between apo and complexed structures of bothropstoxin-I reveals the role of Lys122 and Ca2+-binding loop region for the catalytically inactive Lys49-PLA(2)s

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The Intriguing Phospholipases A(2) Homologues: Relevant Structural Features on Myotoxicity and Catalytic Inactivity

Phospholipases A(2) homologues are found in the venom of Crotalinae snakes, being their main action related to myonecrosis induction. Although many studies on these toxins had already been performed, their mechanism of action remains unclear. Here, important aspects about these toxins are reviewed, including their correct biological assembly and how essential is the natural substitution D49K for their catalytic inactivity.

Morphological, morphometric, and molecular characterization of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from naturally infected Caudisona durissa terrifica (Serpentes, Viperidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Megaselia scalaris (Diptera : Phoridae) causing myiasis in Crotalus durissus terrificus (Serpentes : Viperidae) in Brazil

We describe a case of myiasis in Crotalus durissus terrificus (Laurenti) caused by Megaselia scalaris (Loew). The snake was Found in Anhembi, São Paulo, Brazil, with a lesion measuring 25 mm in diameter where the larvae of M. scalaris had penetrated the ribs. The opportunistic behavior of the larvae of M. scalaris is discussed.

Helminths From an Introduced Species (Tupinambis merianae), and Two Endemic Species (Trachylepis atlantica and Amphisbaena ridleyi) From Fernando de Noronha Archipelago, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Reptilia, Squamata, Dipsadidae, Xenodontinae, Taeniophallus brevirostris (Peters, 1863): distribution extension and new state record.

Taeniophallus brevirostris is a dipsadid snake widely distributed in Amazon. Here we report a new Brazilian state record for the species, improving our knowledge of its geographic distribution. 0n 15 May of 2008, one female specimen was collected do Serra do Santo Expediot, Aripuana municipality, state of Mato Grosso, Brazil. This is the first record of this species for the state of Mato Grosso and extends its distribution ca. 300 kilometers from the nearest localities presented in literature.

Squamata, Elapidae, Micrurus surinamensis (Cuvier, 1817): new records and distribution map in the state of Mato Grosso, Brazil, with notes on diet and activity period.

We report six new records of M. surinamensis and present the map of distribution with known records for this species of semiaquatic coral snake in the state of Mato Grosso, Brazil.

Pulmonary receptors in reptiles: discharge patterns of receptor populations in snakes versus turtles

This study examines the effects of lung inflation/deflation with and without CO2 on the entire population of pulmonary receptors in the vagus nerve in two species of snakes and two species of turtles. We asked the question, how does the response of the entire mixed population of pulmonary stretch receptors (PSR) and intrapulmonary chemoreceptors (IPC) in species possessing both differ from that in species with only PSR? This was studied under conditions of artificial ventilation with the secondary goal of extending observations on the presence/absence of IPC to a further three species. Our results indirectly illustrate the presence of IPC in the Burmese python and South American rattlesnake but not the side necked turtle, adding support to the hypothesis that IPC first arose in diapsid reptiles. In both species of snake, CO2-sensitive discharge (presumably from IPC) predominated almost to the exclusion of CO2-insensitive discharge (presumably arising from PSR) while the opposite was true for both species of turtle. The data suggest that for animals breathing air under conditions of normal metabolism there is little to distinguish between the discharge profiles of the total population of receptors arising from the lungs in the different groups. Interestingly, however, under conditions of elevated environmental CO2 most volume-related feedback from the lungs is abolished in the two species of snakes, while under conditions of elevated metabolic CO2, it is estimated that volume feedback from the lungs would be enhanced in these same species.

The ventilatory response to environmental hypercarbia in the South American rattlesnake, Crotalus durissus

To study the effects of environmental hypercarbia on ventilation in snakes, particularly the anomalous hyperpnea that is seen when CO(2) is removed from inspired gas mixtures (post-hypercapnic hyperpnea), gas mixtures of varying concentrations of CO(2) were administered to South American rattlesnakes, Crotalus durissus, breathing through an intact respiratory system or via a tracheal cannula by-passing the upper airways. Exposure to environmental hypercarbia at increasing levels, up to 7% CO(2), produced a progressive decrease in breathing frequency and increase in tidal volume. The net result was that total ventilation increased modestly, up to 5% CO(2) and then declined slightly on 7% CO(2). on return to breathing air there was an immediate but transient increase in breathing frequency and a further increase in tidal volume that produced a marked overshoot in ventilation. The magnitude of this post-hypercapnic hyperpnea was proportional to the level of previously inspired CO(2). Administration of CO(2) to the lungs alone produced effects that were identical to administration to both lungs and upper airways and this effect was removed by vagotomy. Administration of CO(2) to the upper airways alone was without effect. Systemic injection of boluses of CO(2)-rich blood produced an immediate increase in both breathing frequency and tidal volume. These data indicate that the post-hypercapnic hyperpnea resulted from the removal of inhibitory inputs from pulmonary receptors and suggest that while the ventilatory response to environmental hypercarbia in this species is a result of conflicting inputs from different receptor groups, this does not include input from upper airway receptors.

Meal size and specific dynamic action in the rattlesnake Crotalus durissus (Serpentes : Viperidae)

We studied the effect of meal size on specific dynamic action (SDA) in the South American rattlesnake Crotalus durissus, by measuring oxygen consumption rates (VO2) prior to and after the ingestion of meals ranging from 10-50% of snake's body mass. Regardless of meal size, variation in VO2 with time during digestion demonstrated the same general pattern. Oxygen consumption rates peaked between 15 and 33 h post-feeding, at values 3.7-7.3 times those prior to feeding. Snakes, while digesting meals of 30% and 50% of their body mass, experienced VO2 that exceeded rates measured during forced activity. Following peaks in VO2, rates returned to prefeeding values within 62-170 h post-feeding. Post-prandial peak in VO2 and the duration of the metabolic response to feeding increased with meal size. Digestion is an energetically demanding activity for C. durissus, with an estimated cost equaling 12-18% of the ingested assimilated energy.

Diet of the rattlesnake Crotalus durissus in southeastern Brazil (Serpentes, Viperidae)

Gut contents of 633 live rattlesnakes from southeastern Brazil received at the Instituto Butantanitute, SP, Brazil between 1993 and 1995 were studied. The snakes were weighed, measured and sexed. Two hundred and fifty-nine rattlesnakes had stomach and/or intestinal contents. Prey size was estimated by comparison of prey items with specimens from museum collections. Rodents and small marsupials were the main prey eaten by the rattlesnakes, and only 1% of the items were found in the stomach, whereas 41% of the individuals in the sample had feces in the intestine. There was low correlation between size of snake and prey size. No seasonal difference in frequency was found between fed and not fed males, but the occurrence of fed females was significantly lower during summer than winter months (28.9% and 51.8%, respectively). Fed newborn rattlesnakes had the lowest frequency, and also fed on rodents.

Red blood cells from the South American rattlesnake (Crotalus durissus terrificus) regulate volume incompletely following osmotic shrinkage and swelling in vitro

The ability of rattlesnake (Crotalus durissus terrificus) red blood cells to volume regulate in vitro has been investigated. Blood was drawn through a catheter inserted in the dorsal aorta and equilibrated to gas mixtures of different composition. Cells shrunken osmotically by increasing the extracellular osmolarity from approximate to 291 mosm l(-1) (n = 3) to approximate to 632 mosm l(-1) (calculated) only partially regulated their volume back towards the original volume either at pH 7.51 +/- 0.05 (mean +/- S.D., n = 5) or pH 7.20 +/- 0.06 (mean +/- S.D., n = 3), There was no improvement of the regulatory volume increase at low haemoglobin oxygen saturation. The limited volume restoration was inhibited by separate additions of amiloride (10(-4) M) or DIDS (10(-4) M) suggesting involvement of the Na+/H+ and Cl-/HCO3- exchangers. Cells that were swollen osmotically by an approximate to 30% dilution of the extracellular medium also exhibited a limited ability to recover their volume. Therefore, these cells show little ability to volume regulate when exposed to in vitro conditions that shrink or swell the cell. (C) 2000 Elsevier B.V. All rights reserved.

Effects of temperature on lung and blood gases in the South American rattlesnake Crotalus durissus terrificus

The effects of temperature on lung and blood gases were measured in the South American rattlesnake (Crotalus durissus terrificus). Arterial blood and lung gas samples were obtained from chronically cannulated animals at 15, 25, and 35 degrees C. As expected for reptiles, arterial pH fell with increased temperature (0.018 U degrees C-1 between 15 and 25 degrees C and 0.011 U degrees C-1 between 25 and 35 degrees C) while lung gas PCO2 rose from 5.8 mmHg at 15 degrees C to 13.2 mmHg at 35 degrees C. Concurrently, lung gas PO2 declined from 132 mmHg at 15 degrees C to 120 mmHg at 35 degrees C, and arterial PO2 increased from 33 to 76 mmHg in that temperature range. Arterial haemoglobin O-2 saturation rose from 0.53 at 15 degrees C to 0.83 at 25 degrees C but became slightly reduced (0.77) with a further elevation of temperature to 35 degrees C. Arterial haemoglobin concentration increased from 1.96 to 2.53 mM between 15 and 35 degrees C, consistent with higher demands on oxygen delivery to tissues at elevated temperatures. Moreover, the substantial increase of haemoglobin O-2 saturation between 15 and 25 degrees C conforms to the idea that reduction of the central vascular right-to-left shunt (pulmonary bypass of systemic venous return) is associated with high metabolic demands. (C) 1998 Elsevier B.V. All rights reserved.