Untangling interactions between Bitis vipers and their prey using coagulotoxicity against diverse vertebrate plasmas.

Venom is a key evolutionary innovation which plays a primary role in prey subjugation by venomous snakes. However, while there is a growing body of literature indicating the composition and activity of snake venoms is under strong natural selection driven by differences in prey physiology, the majority of studies have historically focussed on the activity of snake venoms with regards only towards human or mammalian physiologies. This study aimed to use clotting assays measuring both time and strength of clotting to characterise the coagulotoxic activity of venoms from a taxonomically, morphologically, and ecologically diverse range of Bitis spp. of viperid snakes upon the plasma of model species: amphibian (Cane Toad, Rhinella marina); lizard (Blue-tongue Skink, Tiliqua scincoides); avian (Domestic Chicken, Gallus gallus); and rodent (Brown Rat, Rattus norvegicus). Significant variation in coagulotoxic activity across the different plasmas was observed between species and compared to the known affects upon human plasma. Bitis caudalis was notable in being active on all four plasmas, but in extremely divergent manners: accelerating clotting times and producing strong, stable clots upon amphibian plasma (consistent with true procoagulation); accelerating clotting time but producing weak, unstable clots upon lizard plasma (consistent with pseudo-procoagulation); delaying avian clotting time beyond machine maximum reading time (strong anticoagulation consistent with either inhibition of clotting enzymes or total destruction of fibrinogen, or both); and delaying clotting of rodent plasma (consistent with inhibition of clotting enzymes) and with only weak clots formed (consistent with destruction of fibrinogen). In contrast, the sister species B. peringueyi and B. schneideri displayed activity only upon the lizard plasma, slightly accelerating the clotting times to produce weak, unstable clots (consistent with pseudo-procoagulation). The other dwarf species, B. cornuta, displayed strong anticoagulation upon avian and rodent plasmas, delaying clotting beyond the machine maximum reading time (strong anticoagulation consistent with either inhibition of clotting enzymes or total destruction of fibrinogen, or both). In contrast, the giant species studied (B. gabonica) showed only a very weak pseudo-procoagulant activity upon lizard plasma. The wide range of variation seen within this study highlights the importance of studying venom activity on relevant models when making conclusions about the ecological role of venoms and the extreme limitation in extrapolating animal results to predict potential human clinical effects.

Differential coagulotoxic and neurotoxic venom activity from species of the arboreal viperid snake genus Bothriechis (palm-pitvipers).

The viperid snake genus Bothriechis consists of eleven species distributed among Central and South America, living across low and high-altitude habitats. Despite Bothriechis envenomations being prominent across the Central and South American region, the functional effects of Bothriechis venoms are poorly understood. Thus, the aim of this study was to investigate the coagulotoxic and neurotoxic activities of Bothriechis venoms to fill this knowledge gap. Coagulotoxic investigations revealed Bothriechis nigroviridis and B. schlegelii to have pseudo-procoagulant venom activity, forming weak clots that rapidly break down, thereby depleting fibrinogen levels and thus contributing to a net anticoagulant state. While one sample of B. lateralis also showed weaker pseudo-procoagulant activity, directly clotting fibrinogen, two samples of B. lateralis venom were anticoagulant through the inhibition of thrombin and factor Xa activity. Differential efficacy of PoliVal-ICP antivenom was also observed, with the pseudo-procoagulant effect of B. nigroviridis venom poorly neutralised, despite this same activity in the venom of B. schlegelii being effectively neutralised. Significant specificity of these fibrinogen cleaving toxins was also observed, with no activity upon model amphibian, avian, lizard or rodent plasma observed. However, upon avian plasma the venom of B. nigroviridis exerted a complete anticoagulant effect, in contrast to the pseudo-procoagulant effect seen on human plasma. Neurotoxic investigations revealed B. schlegelii to be unique among the genus in having potent binding to the orthosteric site of the alpha-1 postsynaptic nicotinic acetylcholine receptor (with B. lateralis having a weaker but still discernible effect). This represents the first identification of postsynaptic nAChR neurotoxic activity for Bothriechis. In conclusion this study identifies notable differential activity within the coagulotoxic and postsynaptic neurotoxic activity of Bothriechis venoms, supporting previous research, and highlights the need for further studies with respect to antivenom efficacy as well as coagulotoxin specificity for Bothriechis venoms.

Evidence for Resistance to Coagulotoxic Effects of Australian Elapid Snake Venoms by Sympatric Prey (Blue Tongue Skinks) but Not by Predators (Monitor Lizards).

Some Australian elapids possess potently procoagulant coagulotoxic venoms which activate the zymogen prothrombin into the functional enzyme thrombin. Although the activity of Australian elapid prothrombin-activators has been heavily investigated with respect to the mammalian, and in particular, human clotting cascades, very few studies have investigated the activity of their venom upon reptile plasmas. This is despite lizards representing both the primary diet of most Australian elapids and also representing natural predators. This study investigated the procoagulant actions of a diverse range of Australian elapid species upon plasma from known prey species within the genera Tiliqua (blue tongue skinks) as well as known predator species within the genera Varanus (monitor lizards). In addition to identifying significant variation in the natural responses of the coagulation cascade between species from the genera Tiliqua and Varanus relative to each other, as well as other vertebrate lineages, notable differences in venom activity were also observed. Within the genus Tiliqua, both T. rugosa and T. scincoides plasma displayed significant resistance to the procoagulant activity of Pseudechis porphyriacus venom, despite being susceptible to all other procoagulant elapid venoms. These results indicate that T. rugosa and T. scincoides have evolved resistance within their plasma to the coagulotoxic venom activity of the sympatric species P. porphyriacus. Other venoms were able to activate Tiliqua prothrombin, which suggests that the lessened activity of P. porphyriacus venom is not due to modifications of the prothrombin and may instead be due to a serum factor that specifically binds to P. porphyriacus toxins, as has been previously seen for squirrels resistant to rattlesnake venom. In contrast, none of the predatory lizards studied (Varanus giganteus, V. mertensi and V. varius) demonstrated resistance to the venom. This suggests that the mechanical protection afforded by thick osteodermic scales, and prey handling behaviour, removes a selection pressure for the evolution of resistance in these large predatory lizards. These results therefore reveal differential interactions between venoms of snakes with sympatric lizards that are on opposite sides of the predator-prey arms race.

Recommended Health Care and Disease-Prevention Programs for Herds/Flocks of Exotic Animals.

Preventative health care is an essential part of the ownership and veterinary management of exotic animals. This article provides an overview of the current recommendations for health care and disease-prevention programs for herds or flocks of exotic animals, specifically companion and aviary birds, backyard poultry; snakes, lizards, chelonians, and amphibians; rabbits, ferrets, and common exotic small mammals. Husbandry practices, disease screening suggestions and techniques, and vaccination strategies, where appropriate, are reviewed.

MICROSPORIDIA-ASSOCIATED GRANULOMATOUS DISEASE IN TWO AUSTRALIAN COLLECTIONS OF CAPTIVE INLAND BEARDED DRAGONS (POGONA VITTICEPS).

Microsporidia are obligate, intracellular fungi. In reptiles, they are most commonly reported in squamates. We report the first detection of microsporidiosis in inland bearded dragons (Pogona vitticeps) from Australia, and for the first time, mixed infections of microsporidium and adenovirus in asymptomatic inland bearded dragons. In one collection there were five individuals, one of which was lethargic, inappetent, and had lost weight. Two large ovarian granulomas were palpated (42 × 23 mm and 26 × 19 mm) and were surgically removed. This animal died shortly after surgery. Histological evaluation of these granulomas revealed granulomatous inflammation within or adjacent to ovarian tissue, containing numerous aggregates of microorganisms consistent with microsporidia. The organisms were confirmed as Encephalitozoon pogonae by polymerase chain reaction (PCR) and sequencing. Agamid adenovirus-1 was also detected. These two infectious agents were also detected by PCR in all the other bearded dragons in this collection (n = 5), all of which were asymptomatic. A single dragon from a second collection presented for a routine wellness examination after the sudden death of another dragon in the collection. This dragon had similar intracelomic masses to the dragon from the first collection. These were removed surgically, but the dragon died 5 wk later following 3 wk of treatment with 25 mg/kg fenbendazole PO q7 days. Necropsy samples were collected and the microsporidian Encephalitozoon pogonae was detected in oral-cloacal swabs, blood, and multiple tissues by PCR and sequencing. Agamid adenovirus-1 was not detected in this dragon.

Pets in peril: The relative susceptibility of cats and dogs to procoagulant snake venoms.

Snakebite is a common occurrence for pet cats and dogs worldwide and can be fatal. In Australia the eastern brown snake (Pseudonaja textilis) is responsible for an estimated 76% of reported snakebite cases to domestic pets nationally each year, with the primary pathology being venom-induced consumptive coagulopathy. While only 31% of dogs survive P. textilis bites without antivenom, cats are twice as likely to survive bites (66%). Even with antivenom treatment, cats have a significantly higher survival rate. The reason behind this disparity is unclear. Using a coagulation analyser (Stago STA R Max), we tested the relative procoagulant effects of P. textilis venom-as well as 10 additional procoagulant venoms found around the world-on cat and dog plasma in vitro, as well as on human plasma for comparison. All venoms acted faster upon dog plasma than cat or human, indicating that dogs would likely enter coagulopathic states sooner, and are thus more vulnerable to procoagulant snake venoms. The spontaneous clotting time (recalcified plasma with no venom added) was also substantially faster in dogs than in cats, suggesting that the naturally faster clotting blood of dogs predisposes them to being more vulnerable to procoagulant snake venoms. This is consistent with clinical records showing more rapid onset of symptoms and lethal effects in dogs than cats. Several behavioural differences between cats and dogs are also highly likely to disproportionately negatively affect prognosis in dogs. Thus, compared to cats, dogs require earlier snakebite first-aid and antivenom to prevent the onset of lethal venom effects.