Challenges of regulating commercial use of marine elapid snakes in the Indo-Pacific.

Marine elapid snakes are a diverse, predominantly Indo-West Pacific species group. The persistent removal of some species has an unquantified but potentially dire impact on populations. We conducted the first comprehensive review of the trade in marine elapid snakes based on published literature (1974-2022) and trade data from the only species (i.e., Hydrophis [Lapemis] curtus) whose trade is monitored internationally. Some species and populations were subjected to targeted harvest for their meat and skins for at least the last century; fisheries are possibly the most significant threat to populations of marine elapids, with the highest numbers being exploited either accidentally, incidentally, or opportunistically in Southeast Asian fisheries targeting other seafood, including demersal trawl and squid fisheries. Southeast Asia is the core region for exploitation of marine elapids. Annual offtake is >225,000 individuals of at least 8 species in the Gulf of Thailand. Of 72 recognized marine elapids (all non-CITES [Convention on International Trade in Endangered Species of Wild Fauna and Flora] species), Hydrophis curtus and Hydrophis cyanocinctus dominate the skin trade. Skins of H. curtus are traded mainly in East and Southeast Asia and, to some extent, Europe. Despite some baseline information on the trade of these species, the sustainability of their harvests, particularly in the context of the burgeoning and unmanaged nature of fisheries in the region, remains the major challenge. In an era of declining fish stocks, there has been an increasing trend to commercialize the harvest and use marine elapids that were once considered accidental bycatch and discarded. This trend will continue to pose a significant risk to these snakes unless appropriate fisheries and trade regulations are enforced. Applying the precautionary principle to prevent the overexploitation of sea snakes is an indispensable measure in which trade in regional populations should be regulated through CITES. Accordingly, management plans to identify core distribution regions of exploited species would be crucial for assigning national responsibilities to sustain species and populations in the long term.

Retos para la regulación del uso comercial de serpientes elápidas marinas en el Indo­Pacífico Resumen Las serpientes elápidas marinas son un grupo diverso de especies, predominante en el Indo­Pacífico Occidental. La eliminación persistente de algunas especies tiene un impacto no cuantificado pero potencialmente negativo sobre las poblaciones. Realizamos la primera revisión exhaustiva del comercio de serpientes elápidas marinas con base en la bibliografía publicada (1974­2022) y en los datos comerciales de la única especie (Hydrophis [Lapemis] curtus) cuyo mercado tiene monitoreo internacional. Algunas especies y poblaciones fueron objeto de capturas selectivas por su carne y pieles durante al menos el siglo pasado, las pesquerías son posiblemente la amenaza más importante para las poblaciones de elápidos marinos, ya que el mayor número se explota de forma accidental, incidental u oportunista en las pesquerías del sudeste asiático enfocadas en otros mariscos, incluidas las pesquerías demersales de arrastre y de calamar. El sudeste asiático es la principal región de explotación de elápidos marinos. La captura anual es >225,000 individuos de al menos ocho especies en el Golfo de Tailandia. De los 72 elápidos marinos reconocidos (ninguna especie está en CITES [Convención sobre el Comercio Internacional de Especies Amenazadas de Fauna y Flora Silvestres]), Hydrophis curtus e H. cyanocinctus dominan el mercado de pieles. La piel de H. curtus se comercializa principalemnte en el este y sudeste asiático y, hasta cierto punto, en Europa. Aunque se dispone de cierta información de referencia sobre el comercio de estas especies, la sostenibilidad de sus capturas, sobre todo en el contexto del auge y la falta de gestión de la pesca en la región, sigue siendo el principal reto. En una época de disminución de las poblaciones de peces, ha aumentado la tendencia a comercializar la captura y el uso de elápidos marinos que antes se consideraban capturas accidentales y se descartaban. Esta tendencia seguirá representando un riesgo importante para estas serpientes a menos que se apliquen las regulaciones pesqueras y comerciales adecuadas. La aplicación del principio de precaución para evitar la sobreexplotación de las serpientes marinas es una medida indispensable para regular el comercio de las poblaciones regionales a través de CITES. Por lo tanto, los planes de gestión para identificar las regiones núcleo de distribución de las especies explotadas serían cruciales para asignar responsabilidades nacionales para mantener las especies y las poblaciones a largo plazo.

Morphology, Diet, and Reproduction of Coastal Hydrophis Sea Snakes (Elapidae: Hydrophiinae) at Their Northern Distribution Limit.

The Ryukyu Archipelago represents the northern distribution limit for hydrophiine sea snakes, the largest group of marine reptiles. Ryukyuan sea snakes may have developed distinct local adaptations in morphology and ecology, but they have been poorly studied. We examined preserved specimens of 111 Hydrophismelanocephalusand 61 Hydrophis ornatusfrom the Ryukyu Archipelago to obtain data on morphology, diet, and reproduction. Sexual size dimorphism was detected in H. melanocephalus (mean ± standard deviation of adult snout-vent length: SVL, females 1062 ± 141 mm vs. males 959 ± 96 mm) but not in H. ornatus. Female H. melanocephalus had larger head widths and shorter tail lengths relative to SVL compared to males. Relative girth was low in neonates of both species (1.0-1.3), but increased in adults to about 1.7-2.6 in H. melanocephalus and 1.3-1.8 in female H. ornatus. Stomach contents of H. melanocephalus consisted of ophichthid and congrid eels, a sand diver, and gobies, whereas in H. ornatus, gobies and a goat fish were found. Litter size of three reproductive H. melanocephalus ranged from five to seven, and parturition seems to occur from August to October. Litter size of six H. ornatus ranged from two to seven, and was correlated with maternal SVL. Parturition in H. ornatus probably occurs around November. Different selective forces related to locomotion, feeding and predation risk, which influence the pregnant mother and neonates, may have resulted in having few, long but slender offspring that show positive allometric growth in hind-body girth.

Carbon monoxide inhibits hemotoxic activity of Elapidae venoms: potential role of heme.

Envenomation by hemotoxic enzymes continues to be a major cause of morbidity and mortality throughout the world. With regard to treatment, the gold standard to abrogate coagulopathy caused by these venoms is still the administration of antivenom; however, despite antivenom therapy, coagulopathy still occurs and recurs. Of interest, this laboratory has demonstrated in vitro and in vivo that coagulopathy inducing venom derived from snakes of the family Viperidae exposed to carbon monoxide (CO) is inhibited, potentially by an attached heme. The present investigation sought to determine if venoms derived from snakes of the Elapidae family (taipans and cobras) could also be inhibited with CO or with the metheme inducing agent, O-phenylhydroxylamine (PHA). Assessing changes in coagulation kinetics of human plasma with thrombelastography, venoms from Elapidae snakes were exposed in isolation to CO (five species) or PHA (one specie) and placed in human plasma to assess changes in procoagulant or anticoagulant activity. The procoagulant activity of two taipan venoms and anticoagulant activity of three cobra venoms were significantly inhibited by CO. The venom of the inland taipan was also inhibited by PHA. In sum, these data demonstrate indirectly that the biometal heme is likely bound to these disparate venoms as an intermediary modulatory molecule. In conclusion, CO may not just be a potential therapeutic agent to treat envenomation but also may be a potential modulator of heme as a protective mechanism for venomous snakes against injury from their own proteolytic venoms.

Regulation of expression of venom toxins: silencing of prothrombin activator trocarin D by AG-rich motifs.

Trocarin D (TroD), a venom prothrombin activator from Tropidechis carinatus, shares similar structure and function with blood coagulation factor Xa [Tropidechis carinatus FX (TrFX) a]. Their distinct physiologic roles are due to their distinct expression patterns. The genes of TroD and TrFX are highly similar, except for promoter and intron 1, indicating that TroD has probably evolved by duplication of FX, the plasma counterpart. The promoter insertion in TroD accounts for the elevated but not venom gland-specific expression. Here we examined the roles of 3 insertions and 2 deletions in intron 1 of TroD in the regulation of expression using luciferase as a reporter. By systematic deletions, we showed that a 209 bp region within the second insertion silences expression in mammalian and unmilked venom gland cells. Through bioinformatics analysis, we identified 5 AG-rich motifs in this region. All except the 5th motif are important for silencing function. YY1, Sp3 and HMGB2 were identified to bind these AG-rich motifs and silence gene expression in mammalian cells. Similar AG-rich motif clusters are also found in other toxin genes but not in their physiologic counterparts. Thus, AG-rich motifs contribute to regulation of expression of TroD, and probably other toxin genes.-Han, S. X., Kwong, S., Ge, R., Kolatkar, P. R., Woods, A. E., Blanchet, G., Kini, R. M. Regulation of expression of venom toxins: silencing of prothrombin activator trocarin D by AG-rich motifs.

Coming up for air: thermal dependence of dive behaviours and metabolism in sea snakes.

Cutaneous gas exchange allows some air-breathing diving ectotherms to supplement their pulmonary oxygen uptake, which may allow prolongation of dives and an increased capacity to withstand anthropogenic and natural threatening processes that increase submergence times. However, little is known of the interplay between metabolism, bimodal oxygen uptake and activity levels across thermal environments in diving ectotherms. Here, we show in two species of sea snake (spine-bellied sea snake, Hydrophis curtus; and elegant sea snake, Hydrophis elegans) that increasing temperature elevates surfacing rate, increases total oxygen consumption and decreases dive duration. The majority of dives observed in both species remained within estimated maximal aerobic limits. While cutaneous gas exchange accounted for a substantial proportion of total oxygen consumption (up to 23%), unexpectedly it was independent of water temperature and activity levels, suggesting a diffusion-limited mechanism. Our findings demonstrate that rising water temperature and a limited capability to up-regulate cutaneous oxygen uptake may compromise the proficiency with which sea snakes perform prolonged dives. This may hinder their capacity to withstand ongoing anthropogenic activities like trawl fishing, and increase their susceptibility to surface predation as their natural environments continue to warm.

Pr-SNTX, a short-chain three-finger toxin from Papuan pigmy mulga snake, is an antagonist of muscle-type nicotinic acetylcholine receptor (α2ßδε).

Three-finger toxins (3FTxs) are one of the major components in snake venoms. In this study, we isolated a cDNA encoding a short-chain 3FTx, Pr-SNTX, from Pseudechis rossignolii. The amino acid sequence of Pr-SNTX is nearly identical to that of its ortholog in Pseudechis australis. Pr-SNTX protein inhibited muscle-type (α2ßδε), but not neuronal α7 nicotinic acetylcholine receptor (nAChR) activity.

Spatial variation in age structure among colonies of a marine snake: the influence of ectothermy.

Several tetrapod lineages that have evolved to exploit marine environments (e.g. seals, seabirds, sea kraits) continue to rely upon land for reproduction and, thus, form dense colonies on suitable islands. In birds and mammals (endotherms), the offspring cannot survive without their parents. Terrestrial colonies contain all age classes. In reptiles (ectotherms), this constraint is relaxed, because offspring are independent from birth. Hence, each age class has the potential to select sites with characteristics that favour them. Our studies of sea snakes (sea kraits) in the lagoon of New Caledonia reveal marked spatial heterogeneity in age structure among colonies. Sea krait colonies exhibit the endothermic 'seal-seabird' pattern (mixed-age classes within populations) only where the lagoon is narrow. Where the lagoon is wide, most snake colonies are comprised primarily of a single age cohort. Nurseries are located near the coast, adult colonies offshore and mixed colonies in-between. We suggest that ectothermy allows individuals to utilize habitats that are best suited to their own ecological requirements, a flexibility not available to endothermic marine taxa with obligate parental care.

Pelagic sea snakes dehydrate at sea.

Secondarily marine vertebrates are thought to live independently of fresh water. Here, we demonstrate a paradigm shift for the widely distributed pelagic sea snake, Hydrophis (Pelamis) platurus, which dehydrates at sea and spends a significant part of its life in a dehydrated state corresponding to seasonal drought. Snakes that are captured following prolonged periods without rainfall have lower body water content, lower body condition and increased tendencies to drink fresh water than do snakes that are captured following seasonal periods of high rainfall. These animals do not drink seawater and must rehydrate by drinking from a freshwater lens that forms on the ocean surface during heavy precipitation. The new data based on field studies indicate unequivocally that this marine vertebrate dehydrates at sea where individuals may live in a dehydrated state for possibly six to seven months at a time. This information provides new insights for understanding water requirements of sea snakes, reasons for recent declines and extinctions of sea snakes and more accurate prediction for how changing patterns of precipitation might affect these and other secondarily marine vertebrates living in tropical oceans.

Trophic divergence despite morphological convergence in a continental radiation of snakes.

Ecological and phenotypic convergence is a potential outcome of adaptive radiation in response to ecological opportunity. However, a number of factors may limit convergence during evolutionary radiations, including interregional differences in biogeographic history and clade-specific constraints on form and function. Here, we demonstrate that a single clade of terrestrial snakes from Australia--the oxyuranine elapids--exhibits widespread morphological convergence with a phylogenetically diverse and distantly related assemblage of snakes from North America. Australian elapids have evolved nearly the full spectrum of phenotypic modalities that occurs among North American snakes. Much of the convergence appears to involve the recurrent evolution of stereotyped morphologies associated with foraging mode, locomotion and habitat use. By contrast, analysis of snake diets indicates striking divergence in feeding ecology between these faunas, partially reflecting regional differences in ecological allometry between Australia and North America. Widespread phenotypic convergence with the North American snake fauna coupled with divergence in feeding ecology are clear examples of how independent continental radiations may converge along some ecological axes yet differ profoundly along others.

Rapid evolution of mimicry following local model extinction.

Batesian mimicry evolves when individuals of a palatable species gain the selective advantage of reduced predation because they resemble a toxic species that predators avoid. Here, we evaluated whether-and in which direction-Batesian mimicry has evolved in a natural population of mimics following extirpation of their model. We specifically asked whether the precision of coral snake mimicry has evolved among kingsnakes from a region where coral snakes recently (1960) went locally extinct. We found that these kingsnakes have evolved more precise mimicry; by contrast, no such change occurred in a sympatric non-mimetic species or in conspecifics from a region where coral snakes remain abundant. Presumably, more precise mimicry has continued to evolve after model extirpation, because relatively few predator generations have passed, and the fitness costs incurred by predators that mistook a deadly coral snake for a kingsnake were historically much greater than those incurred by predators that mistook a kingsnake for a coral snake. Indeed, these results are consistent with prior theoretical and empirical studies, which revealed that only the most precise mimics are favoured as their model becomes increasingly rare. Thus, highly noxious models can generate an 'evolutionary momentum' that drives the further evolution of more precise mimicry-even after models go extinct.

Sea snake harvest in the gulf of Thailand.

Conservation of sea snakes is virtually nonexistent in Asia, and its role in human-snake interactions in terms of catch, trade, and snakebites as an occupational hazard is mostly unexplored. We collected data on sea snake landings from the Gulf of Thailand, a hotspot for sea snake harvest by squid fishers operating out of the ports of Song Doc and Khanh Hoi, Ca Mau Province, Vietnam. The data were collected during documentation of the steps of the trading process and through interviewers with participants in the trade. Squid vessels return to ports once per lunar synodic cycle and fishers sell snakes to merchants who sort, package, and ship the snakes to various destinations in Vietnam and China for human consumption and as a source of traditional remedies. Annually, 82 t, roughly equal to 225,500 individuals, of live sea snakes are brought to ports. To our knowledge, this rate of harvest constitutes one of the largest venomous snake and marine reptile harvest activities in the world today. Lapemis curtus and Hydrophis cyanocinctus constituted about 85% of the snake biomass, and Acalyptophis peronii, Aipysurus eydouxii, Hydrophis atriceps, H. belcheri, H. lamberti, and H. ornatus made up the remainder. Our results establish a quantitative baseline for characteristics of catch, trade, and uses of sea snakes. Other key observations include the timing of the trade to the lunar cycle, a decline of sea snakes harvested over the study period (approximately 30% decline in mass over 4 years), and the treatment of sea snake bites with rhinoceros horn. Emerging markets in Southeast Asia drive the harvest of venomous sea snakes in the Gulf of Thailand and sea snake bites present a potentially lethal occupational hazard. We call for implementation of monitoring programs to further address the conservation implications of this large-scale marine reptile exploitation.

Selection overrides gene flow to break down maladaptive mimicry.

Predators typically avoid dangerous species, and batesian mimicry evolves when a palatable species (the 'mimic') co-opts a warning signal from a dangerous species (the 'model') and thereby deceives its potential predators. Because predators would not be under selection to avoid the model and any of its look-alikes in areas where the model is absent (that is, allopatry), batesian mimics should occur only in sympatry with their model. However, contrary to this expectation, batesian mimics often occur in allopatry. Here we focus on one such example--a coral snake mimic. Using indirect DNA-based methods, we provide evidence suggesting that mimics migrate from sympatry, where mimicry is favoured, to allopatry, where it is disfavoured. Such gene flow is much stronger in nuclear genes than in maternally inherited mitochondrial genes, indicating that dispersal by males may explain the presence of mimetic phenotypes in allopatry. Despite this gene flow, however, individuals from allopatry resemble the model less than do individuals from sympatry. We show that this breakdown of mimicry probably reflects predator-mediated selection acting against individuals expressing the more conspicuous mimetic phenotype in allopatry. Thus, although gene flow may explain why batesian mimics occur in allopatry, natural selection may often override such gene flow and promote the evolution of non-mimetic phenotypes in such areas.

Recent rapid speciation and ecomorph divergence in Indo-Australian sea snakes.

The viviparous sea snakes (Hydrophiinae) are a young radiation of at least 62 species that display spectacular morphological diversity and high levels of local sympatry. To shed light on the mechanisms underlying sea snake diversification, we investigated recent speciation and eco-morphological differentiation in a clade of four nominal species with overlapping ranges in Southeast Asia and Australia. Analyses of morphology and stomach contents identified the presence of two distinct ecomorphs: a 'macrocephalic' ecomorph that reaches >2 m in length, has a large head and feeds on crevice-dwelling eels and gobies; and a 'microcephalic' ecomorph that rarely exceeds 1 m in length, has a small head and narrow fore-body and hunts snake eels in burrows. Mitochondrial sequences show a lack of reciprocal monophyly between ecomorphs and among putative species. However, individual assignment based on newly developed microsatellites separated co-distributed specimens into four significantly differentiated clusters corresponding to morphological species designations, indicating limited recent gene flow and progress towards speciation. A coalescent species tree (based on mitochondrial and nuclear sequences) and isolation-migration model (mitochondrial and microsatellite markers) suggest between one and three transitions between ecomorphs within the last approximately 1.2 million to approximately 840,000 years. In particular, the macrocephalic 'eastern' population of Hydrophis cyanocinctus and microcephalic H. melanocephalus appear to have diverged very recently and rapidly, resulting in major phenotypic differences and restriction of gene flow in sympatry. These results highlight the viviparous sea snakes as a promising system for speciation studies in the marine environment.

Comparison of freshwater discrimination ability in three species of sea kraits (Laticauda semifasciata, L. laticaudata and L. colubrina).

Three species of amphibious sea kraits (Laticauda spp.) require drinking freshwater to regulate water balance. The extent of terrestriality is known to differ among them. Species with higher extent of terrestriality would drink freshwater accumulated on land, whereas less terrestrial species would rely totally on freshwater that runs into the sea. Consequently, we predicted that the latter species might have a better ability to follow the flow of freshwater or lower salinity water in the sea than the former. We investigated the freshwater discrimination ability of three sea krait species, using a Y-maze apparatus. We found that Laticauda semifasciata and Laticauda laticaudata, less terrestrial species, followed freshwater significantly more frequently than seawater, whereas Laticauda colubrina, more terrestrial species, unbiasedly selected freshwater and seawater. This result supports our prediction and suggests that less terrestrial sea kraits more efficiently access freshwater sources in the sea than highly terrestrial sea kraits. It is likely that behavioral rehydration systems vary among sea kraits in relation to their terrestrial tendency.

Potential targets aimed at by spitting cobras when deterring predators from attacking.

When threatened, spitting cobras eject venom towards the face of an aggressor. To uncover the relevant cues used by cobras for face recognition we determined how often artificial targets equipped with or without eyes elicited spitting behavior. In addition, we measured whether and how target shape and size influenced the spitting behavior of cobras. Results show that oval- and round-shaped targets were most effective, while triangles with the same surface area as oval 'face like' targets hardly elicited spitting. The likelihood of spitting depended on neither the presence, the spatial arrangement (horizontal or vertical) nor the surface texture (shiny or matt) of glass eyes. Most likely, cobras do not specifically aim at the eyes of an offender but at the center of the body part closest to them. As this is usually the face of an animal, this strategy will result in at least one eye of the offender being hit most of the time.

Pairwise comparison of orthologous olfactory receptor genes between two sympatric sibling sea kraits of the genus Laticauda in Vanuatu.

Olfaction-based reproductive isolation is widely observed in animals, but little is known about the genetic basis of such isolation mechanisms. Two species of sibling amphibious sea snakes, Laticauda colubrina and L. frontalis live in Vanuatu sympatrically and syntopically, but no natural hybrids have been reported. Adult females of both taxa possess distinctive lipids in the skin, and male L. frontalis distinguishes conspecific females based on olfactory cues. To shed light on the molecular basis of the evolution of olfaction-based isolation mechanisms, olfactory receptor (OR) gene repertoires of both taxa were identified using pyrosequencing-based technology, and orthologous OR gene sets were identified. Few species-specific gene duplications or species-specific gene losses were found. However, the nonsynonymous-to-synonymous substitution rate ratio was relatively higher between orthologous OR genes of L. frontalis and L. colubrina, indicating that L. frontalis and L. colubrina have evolved to possess different olfactory senses. We suggest that L. frontalis and L. colubrina have evolved allopatrically, and this may be a byproduct of the allopatric evolution, and that this dissimilarity may function as a premating isolation barrier, since L. frontalis has returned to the ancestral range (Vanuatu).

Geographic genetic structure in two laticaudine sea kraits, Laticauda laticaudata and Laticauda semifasciata (Serpentes: Elapidae), in the Ryukyu-Taiwan region as inferred from mitochondrial cytochrome b sequences.

The Ryukyu-Taiwan region is an island arch with intervening waters of varying distances and depths. This study examines the geographic genetic structure of two sympatric sea kraits, Laticauda laticaudata and L. semifasciata, in the region, to infer factors affecting the extent of dispersal and other biogeographical traits of these amphibious reptiles. Sequence analyses of the mitochondrial cytochrome b gene revealed four and 16 haplotypes for L. laticaudata (136 individuals) and L. semifasciata (177 individuals), respectively. For both species, population pairwise F ST analyses revealed significant genetic differentiations among islands and island groups, which are separated by deep straits, suggesting that deep waters serve as obstacles for dispersal in both species. Significant genetic differentiation was detected even among islands of the same basin in L. laticaudata, but not in L. semifasciata, and the isolation by distance analyses revealed no significant correlation between geographic and genetic distances in the former species. These results further suggest that L. laticaudata has stronger site fidelity or degree of philopatry than L. semifasciata. Based on the geographic genetic patterns, the historical biogeography of the two species in the Ryukyu-Taiwan region is also discussed.

Structural characteristics and evolution of the Protobothrops elegans pancreatic phospholipase A2 gene in contrast with those of Protobothrops genus venom phospholipase A2 genes.

The nucleotide sequence of the gene encoding Protobothrops elegans (Crotalinae) pancreatic phospholipase A(2) (PLA(2)), abbreviated PePancPLA(2), was determined by means of inverted PCR techniques. Since its deduced amino acid sequence contains a pancreatic loop and shows high similarity to that of Laticauda semifasciata (Elapinae) group IB pancreatic PLA(2), PePancPLA(2) is classified into group IB PLA(2). The nucleotide sequences of the PePancPLA(2) gene, the L. semifasciata group IB pancreatic PLA(2) gene, and the L. semifasciata group IA venom PLA(2) gene are similar to one another but greatly dissimilar to those of Protobothrops genus (Crotalinae) group II venom PLA(2) genes, suggesting that the Elapinae group IB PLA(2) gene and the group IA PLA(2) gene appeared after Elapinae was established, and that the Crotalinae group II venom PLA(2) genes came into existence before Elapinae and Crotalinae diverged. A phylogenetic analysis of their amino acid sequences confirms this.

Cardiotoxin-I: an unexpectedly potent insulinotropic agent.

Insulin secretion from pancreatic ß-cells is a complex process, involving the integration and interaction of multiple external and internal stimuli, in which glucose plays a major role. Understanding the physiology leading to insulin release is a crucial step toward the identification of new targets. In this study, we evaluated the presence of insulinotropic metabolites in Naja kaouthia snake venom. Only one fraction, identified as cardiotoxin-I (CTX-I) was able to induce insulin secretion from INS-1E cells without affecting cell viability and integrity, as assessed by MTT and LDH assays. Interestingly, CTX-I was also able to stimulate insulin secretion from INS-1E cells even in the absence of glucose. Although cardiotoxins have been characterized as potent hemolytic agents and vasoconstrictors, CTX-I was unable to induce direct hemolysis of human erythrocytes or to induce potent vasoconstriction. As such, this newly identified insulin-releasing toxin will surely enrich the pool of existing tools to study ß-cell physiology or even open a new therapeutic avenue.

Therapeutic potential of Naja naja atra venom in a rat model of diabetic nephropathy.

OBJECTIVE: To study the protective effects of naja naja atra venom (NNAV) in a rat model of diabetic nephropathy (DN). METHODS: The rat diabetes model was induced by intraperitoneal injection of streptozotocin (STZ). Thirty-two model rats were randomly divided into one DN group (n=8) and three treatment groups (n=8 each) that received NNAV at doses of 30, 90, or 270 µg/(kg·day) via oral gavage, another eight rats as normal controls. After 12 weeks, all rats were sacrificed and the changes in serum and urine biological index levels were determined by colorimetric assay. Microalbumin (mALB), N-acetyl-ß- glucosaminidase (NAG) and cystatin C (CysC) concentrations were measured by ELISA. Renal tissues were sliced for pathological and immunohistochemical observations. RESULTS: Comparied with the DN group, serum glucose was decreased by 31.04%, total cholesterol 21.96%, triglyceride 23.78%, serum creatinine 19.83%, blood urea nitrogen 31.28%, urinary protein excretion 45.42%, mALB 10.42%, NAG 20.65%, CysC 19.57%, whereas albumin increased by 5.55%, high-density lipoprotein-cholesterol 59.09%, creatinine clearance 19.05% in the treatment group by NNAV administration at dose of 90 µg/(kg·day). NNAV also reduced the levels of malondialdehyde in serum (22.56%) and kidney tissue (9.79%), and increased superoxide dismutase concentration in serum (15%) and decreased it in renal tissue (8.85%). In addition, under light microscopy kidney structure was improved and glomerular hypertrophy decreased by 8.29%. As shown by immunohistochemistry, NNAV inhibited transforming growth factor-ß1 by 6.70% and nuclear actor-κB by 5.15%. CONCLUSION: NNAV improves biological indexes in DN, and it may exert renoprotective effects in rats with STZ-induced diabetes.