The Treatment of Unidentified Hematotoxic Snake Envenomation and the Clinical Manifestations of a Protobothrops kelomohy Bite.

Hematotoxic snake envenomation is clinically important and has serious complications. This case report describes the clinical manifestations of a bite from a newly described species of hematotoxic snake and the outcomes of treatment using locally available antivenom and supportive management. A 39-y-old male was bitten on his right ankle by an unidentified snake. The patient developed coagulopathy, rhabdomyolysis, and clinical signs of compartment syndrome of his right leg within the first day. Eight doses (30 mL, or 3 vials per dose), or a total of 240 mL, of hemato-polyvalent antivenin (Thai Red Cross, Thailand) were given. Aggressive intravenous hydration and alkalinization of urine were performed, and a fasciotomy was performed for the treatment of suspected compartment syndrome. The patient's clinical symptoms and laboratory results indicated progressive improvement during the 5 d of hospitalization. The snake was later identified as Protobothrops kelomohy, a novel snake species with no clinical data available regarding its envenomation of humans. This case shows that P kelomohy envenomation can produce severe effects that occur both locally and systemically. The benefits of the use of polyvalent antivenom in this situation were unclear. However, with supportive treatment in conjunction with serial clinical and laboratory monitoring, the patient recovered, albeit after 7 d of hospitalization.

Phylogeny and toxicological assessments of two Porthidium lansbergii lansbergii morphotypes from the caribbean region of Colombia.

After a snakebite accident, species identification is of vital importance. However, the existence of intraspecific differences in the body coloration patterns of venomous snakes can generate confusion and delay a convenient and effective treatment. This is the situation for Porthidium lansbergii lansbergii from Colombia, for which two distinctive color morphs occur, and the relationship of these morphs with venom toxicity is unknown. Therefore, venom samples from specimens of these two morphs were collected from the Colombian Caribbean region, and their protein profiles compared. Likewise, their venom functional activities were evaluated in vitro and in vivo in BALB/C mice. Additionally, using sequences of the mitochondrial cytochrome b (Cyt-b) gene, the relationship between these Colombian P. lansbergii lansbergii morphotypes was investigated, and their phylogenetic positions were determined for the first time using Bayesian inference. Despite the noticeable coloration divergence between the individuals analyzed, similar protein profiles of their venoms were observed. Additionally, neither their lethality nor biochemical activities were notably different. In general, both venoms were highly proteolytic, lacked a coagulant effect in vitro, and extended the clotting time due to the action of venom components, such as disintegrins and proteases, that induce defibrination. These results agreed with the result of our phylogenetic analysis, suggesting that the two chromatic morphs do not represent isolated populations. The phylogenetic analyses also supported the currently recognized P. lansbergii lansbergii subspecies as a monophyletic complex. In conclusion, the results of this investigation suggest similar clinical manifestations regardless of body coloration after a P. lansbergii lansbergii envenomation, and pools can therefore be used for antivenom development, medical treatments, and further research efforts.

Venom characterization of the three species of Ophryacus and proteomic profiling of O. sphenophrys unveils Sphenotoxin, a novel Crotoxin-like heterodimeric ß-neurotoxin.

Venoms of the three species of Ophryacus (O. sphenophrys, O. smaragdinus, and O. undulatus), a viperid genus endemic to Mexico, were analyzed for the first time in the present work. The three venoms lacked procoagulant activity on human plasma, but induced hemorrhage and were highly lethal to mice. These venoms also displayed proteolytic and phospholipase A2 activities in vitro. The venom of O. sphenophrys was the most lethal and caused hind-limb paralysis in mice. Proteomic profiling of O. sphenophrys venom showed a predominance of metalloproteinase (34.9%), phospholipase A2 (24.8%) and serine protease (17.1%) in its composition. Strikingly, within its PLA2 components, 12.9% corresponded to a Crotoxin-like heterodimer, here named Sphenotoxin, which was not found in the other two species of Ophryacus. Sphenotoxin, like Crotoxin, is composed of non-covalently bound A and B subunits. Partial amino acid sequence was obtained for Sphenotoxin B and was similar (78-89%) to other subunits described. The mouse i.v. LD50 of Sphenotoxin at 1:1 M radio was 0.16 µg/g. Also, like Crotoxin, Sphenotoxin induced a potent neuromuscular blockade in the phrenic nerve-diaphragm preparation. Ophryacus is the fifth genus and O. sphenophrys the third non-rattlesnake species shown to contain a novel Crotoxin-like heterodimeric ß-neurotoxin. BIOLOGICAL SIGNIFICANCE: Ophryacus is an endemic genus of semi-arboreal pitvipers from Mexico that includes three species with restricted distributions. Little is known about the natural history of these species and nothing is known about the properties of their venoms. Research on these species' venoms could generate relevant information regarding venom composition of Mexican pitvipers. Additionally, research into the presence of neurotoxic Crotoxin-like molecules outside of rattlesnakes (genera Crotalus and Sistrurus) has identified this molecule in several new genera. Knowing which genera and species possess neurotoxic components is important to fully understand the repercussions of snakebites, the interaction with prey and predators, and the origin, evolution, and phylogenetic distribution of Crotoxin-like molecules during the evolutionary history of pitvipers. Our study expands current knowledge regarding venom's compositions and function from Mexican pitvipers, providing a comparative venom characterization of major activities in the three Ophryacus species. Additionally, the discovery and characterization of a novel Crotoxin-like molecule, here named Sphenotoxin, in O. sphenophrys, and the detailed protein composition of O. sphenophrys venom supports the hypotheses that Crotoxin-like -ß-neurotoxins are more widespread than initially thought.

Sequencing and analysis of the complete mitochondrial genome of Gloydius saxatilis (Squamata: Viperidae: Crotalinae).

The mitochondrial genome sequence of Gloydius saxatilis is analyzed and presented for the public for the first time. The genome was 17,218 bp in length and contained 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and 2 control regions. The overall base composition was A (32.3%), C (28.9%), T (25.8%), and G (13.0%). The base compositions presented clearly the A-C skew, which was most obviously in the protein-coding genes. The extended termination-associated sequence domain, the central conserved domain and the conserved sequence block domain are defined in the mitochondrial genome control region of G. saxatilis. Mitochondrial genomes analyses based on MP, ML, NJ and Bayesian analyses yielded identical phylogenetic trees, indicating a close phylogenetic affinity of the thirteen Crotalinae species. It appeared that no less than two major phyletic lineages were present in Crotalinae. The main clades within the Crotalinae supported are: A clade including the Protobothrops. A clade (G. brevicaudus, G. ussuriensis, G. intermedius, G. saxatilis) with the Ovophis as the sister taxon to Protobothrops and was supported by bootstrap values of 88%. The four Gloydius species formed a paraphyletic group with the high bootstrap value (100%) in all examinations.

The complete mitochondrial genome of Gloydius intermedius (Squamata: Viperidae: Crotalinae) from China.

The mitochondrial genome sequence of Gloydius intermedius is analyzed and presented for the first time. The genome was 17, 226 bp in length and contained 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and 2 control region. The overall base composition was A (32.4%), C (28.8%), T (25.9%), and G (12.9%). The base compositions clearly presented the A-C skew, which was most obvious in the protein-coding genes. The extended termination-associated sequence domain, the central conserved domain and the conserved sequence block domain are defined in the mitochondrial genome control region of G. intermedius. Mitochondrial genome analyses based on MP, ML, NJ and Bayesian analyses yielded identical phylogenetic trees, indicating a close phylogenetic affinity of the 13 Crotalinae species. It appeared that no less than two major phyletic lineages were present in Crotalinae. The main clades within the Crotalinae supported are: A clade including the Protobothrops. A clade (G. brevicaudus, G. ussuriensis, G. intermedius, G. saxatilis) with the Ovophis appeared as the sister taxon to Protobothrops and was supported by bootstrap values of 88%. The four Gloydius species formed a paraphyletic group with the high bootstrap value (100 %) in all examinations.

The complete mitochondrial genome sequence of Gloydius shedaoensis (Squamata: Viperidae).

Gloydius shedaoensis is an insular and vulnerable pitviper that is endemic to Snake Island, northeastern China. In this study, we successfully sequenced mitochondrial genomes of two individuals of G. shedaoensis. The complete mitochondrial genomes of G. shedaoensis are circular molecular with 17 222 and 17 221 bp in length respectively, which both contain 2 ribosomal RNA (rRNA) genes, 13 protein-coding genes, 22 transfer RNA (tRNA) genes, an origin of light-strand replication (OL) and two non-coding control regions. Compared with previously published mitochondrial genomes of Gloydius species, the base composition and gene arrangement are rather conservative. A Bayesian phylogenetic tree using the complete mitochondrial genomes of all viper species available showed a consistent result with previous studies.