Venomous Snake Species Research Articles

Protein structure of the venom in nine species of snake: from bio-compounds to possible healing agents.

Due to its various structures in bio-compounds, snake venom is the indisputable result of evolutionary stages of molecules with an increasingly complex structure, high specificity, and of great importance for medicine because of their potential. The present study proposed an underpinning examination of venom composition from nine species of venomous snakes using a useful and replicable methodology. The objective was the extension of the evaluation of protein fractions in the field up to 230 kDa to permit possible identification of some fractions that are insufficiently studied. The gel capillary electrophoresis method on the chip was performed using an Agilent 2100 bioassay with the 80 and 230-LabChip Protein kits. Interpretation of electrophoresis was performed using the Protein 2100 expert (Agilent) test software as follows: a) Protein 80 (peak size scale): 1.60, 3.5, 6.50, 15.00, 28.00, 46.00, 63.00, 95.00 kDa; b) Protein 230 (peak size scale): 4.50, 7.00, 15.00, 28.00, 46.00, 63.00, 95.00, 150.00, 240.00 kDa. The screening revealed the presence of compounds with a molecular weight greater than 80 kDa, in the case of Vipera aspis and Vipera xantina palestinae. For V. aspis, a 125 kDa molecular weight pro-coagulant protein was identified, known as being involved in the reduction of plasma clotting time without any direct activity in the fibrinogen coagulation process. The samples examined on the Protein 230-LabChip electrophoresis chip can be considered as a novelty with possible uses in medicine, requiring further approaches by advanced proteomics techniques to confirm the intimate structural features and biological properties of snake venoms.

Crotamine in Crotalus durissus: distribution according to subspecies and geographic origin, in captivity or nature.

Background: Crotalus durissus is considered one of the most important species of venomous snakes in Brazil, due to the high mortality of its snakebites. The venom of Crotalus durissus contains four main toxins: crotoxin, convulxin, gyroxin and crotamine. Venoms can vary in their crotamine content, being crotamine-negative or -positive. This heterogeneity is of great importance for producing antivenom, due to their different mechanisms of action. The possibility that antivenom produced by Butantan Institute might have a different immunorecognition capacity between crotamine-negative and crotamine-positive C. durissus venoms instigated us to investigate the differences between these two venom groups. Methods:The presence of crotamine was analyzed by SDS-PAGE, western blotting and ELISA, whereas comparison between the two types of venoms was carried out through HPLC, mass spectrometry analysis as well as assessment of antivenom lethality and efficacy. Results:The results showed a variation in the presence of crotamine among the subspecies and the geographic origin of snakes from nature, but not in captive snakes. Regarding differences between crotamine-positive and -negative venoms, some exclusive proteins are found in each pool and the crotamine-negative pool presented more phospholipase A2 than crotamine-positive pool. This variation could affect the time to death, but the lethal and effective dose were not affected.Conclusion:These differences between venom pools indicate the importance of using both, crotamine-positive and crotamine-negative venoms, to produce the antivenom.

Beyond the 'big four': Venom profiling of the medically important yet neglected Indian snakes reveals disturbing antivenom deficiencies.

Snakebite in India causes the highest annual rates of death (46,000) and disability (140,000) than any other country. Antivenom is the mainstay treatment of snakebite, whose manufacturing protocols, in essence, have remained unchanged for over a century. In India, a polyvalent antivenom is produced for the treatment of envenomations from the so called 'big four' snakes: the spectacled cobra (Naja naja), common krait (Bungarus caeruleus), Russell's viper (Daboia russelii), and saw-scaled viper (Echis carinatus). In addition to the 'big four', India is abode to many other species of venomous snakes that have the potential to inflict severe clinical or, even, lethal envenomations in their human bite victims. Unfortunately, specific antivenoms are not produced against these species and, instead, the 'big four' antivenom is routinely used for the treatment. We characterized the venom compositions, biochemical and pharmacological activities and toxicity profiles (mouse model) of the major neglected yet medically important Indian snakes (E. c. sochureki, B. sindanus, B. fasciatus, and two populations of N. kaouthia) and their closest 'big four' congeners. By performing WHO recommended in vitro and in vivo preclinical assays, we evaluated the efficiencies of the commercially marketed Indian antivenoms in recognizing venoms and neutralizing envenomations by these neglected species. As a consequence of dissimilar ecologies and diet, the medically important snakes investigated exhibited dramatic inter- and intraspecific differences in their venom profiles. Currently marketed antivenoms were found to exhibit poor dose efficacy and venom recognition potential against the 'neglected many'. Premium Serums antivenom failed to neutralise bites from many of the neglected species and one of the 'big four' snakes (North Indian population of B. caeruleus). This study unravels disturbing deficiencies in dose efficacy and neutralisation capabilities of the currently marketed Indian antivenoms, and emphasises the pressing need to develop region-specific snakebite therapy for the 'neglected many'.

A comparison of wildlife value orientations and attitudes toward timber rattlesnakes (Crotalus horridus)

ABSTRACTHuman-wildlife interactions can be explained using a cognitive hierarchy model, in which wildlife value orientations (WVO) form the basis for human beliefs, attitudes, and behaviors related to wildlife and wildlife management. In this article, we used data from a mail survey to test the fit of an established WVO framework to a venomous snake species. Study participants were divided into four WVO types (Distanced, Mutualists, Pluralists, Traditionalists). Results indicated that WVO types varied by sex and certain lifestyle factors. Attitudes toward timber rattlesnakes also varied by WVO type. Pluralists exhibited the most favorable attitudes while Traditionalists exhibited the most adverse attitudes toward timber rattlesnakes. Potential for Conflict Index (PCI2) analysis revealed variation among members of each WVO type in support for timber rattlesnake management strategies. This segmentation method can help wildlife managers and researchers better understand the diversity of human characteristics related to conservation and management of wildlife species.

REVIEW OF OPISTHOGLYPHOUS SNAKES (REPTILIA, OPHIDIA ) OF IRAQ

Seven species of semi venomous Opisthoglypha snakes (Reptilia, Ophidia) of Iraq are listed with important characteristics in morphology due to geographical and individual variation of species, as well, the confusion in the scales count of Telescopus tessellatus martini (Schmidt, 1939) of Iraq are discussed.

Recent advances in snake venom proteomics research in India: a new horizon to decipher the geographical variation in venom proteome composition and exploration of candidate drug prototypes

India is the home to several venomous snake species which are responsible for the highest number of snake envenomings and deaths in the world. Nevertheless, the ‘Big Four’ species of snakes namely Russell’s viper (Daboia russelii), spectacled cobra (Naja naja), common krait (Bungarus caeruleus), and saw-scaled viper (Echis carinatus) account for most cases of envenoming, morbidity, and mortality primarily in the rural parts of the country. The composition of these snake venoms have been determined using conventional biochemical approaches. However, these studies faced the inevitable limitation of identification of non-enzymatic and low abundance venom toxins. In the last 5 years, mass spectrometry investigations of Indian snake venoms have provided comprehensive information on their venom proteomes. In addition, proteomic analysis has also been employed to identify and characterize snake venom protein complexes and serves as a tool for screening of candidate pharmacologically active drug prototypes from snake venoms. Further, mass spectrometry analysis has also found its way for the identification and quantification of poorly immunogenic venom toxins against commercial antivenoms and this information is of paramount importance for improved design and production of snake antivenoms for better hospital management of snakebite victims. In this review article, we have focused the recent advances in snake venom proteomics research in India that has facilitated the understanding of the pathophysiology of snake envenomation, provided insights for improvement for antivenom production and also discover drug prototypes from snake venom.

Malaysian Cobra Venom: A Potential Source of Anti-Cancer Therapeutic Agents.

Cancer is a deadly disease and there is an urgent need for the development of effective and safe therapeutic agents to treat it. Snake venom is a complex mixture of bioactive proteins that represents an attractive source of novel and naturally-derived anticancer agents. Malaysia is one of the world’s most biodiverse countries and is home to various venomous snake species, including cobras. Naja kaouthia, Naja sumatrana, and Ophiophagus hannah are three of the most common cobra species in Malaysia and are of medical importance. Over the past decades, snake venom has been identified as a potential source of therapeutic agents, including anti-cancer agents. This present review highlights the potential anticancer activity of the venom and purified venom protein of N. kaouthia, N. sumatrana, and O. hannah. In conclusion, this review highlights the important role of the venom from Malaysian cobras as an important resource that researchers can exploit to further investigate its potential in cancer treatment.

Clinical and laboratory features distinguishing between Deinagkistrodon acutus and Daboia siamensis envenomation

BackgroundThere are 6 species of venomous snakes in Taiwan. Two of them, Deinagkistrodon acutus (D. acutus) and Daboia siamensis (D. siamensis), can cause significant coagulopathy. However, a significant proportion of patients with snakebites cannot identify the correct snake species after envenomation, which hampers the application of antivenom. Hence, the differential diagnosis between the two snakebites by clinical presentations is important. This study aims to compare their clinical and laboratory features for the purpose of differential diagnosis between the two snakebites.MethodsWe retrospectively reviewed the medical records of patients who arrived at the emergency department due to D. acutus or D. siamensis envenomation, between 2003 and 2016, in one medical center in eastern Taiwan. Since these snakebites are rare, we also included 3 cases reported from another hospital in central Taiwan.ResultsIn total, 15 patients bitten by D. acutus and 12 patients by D. siamensis were analyzed. Hemorrhagic bulla formation and the need for surgical intervention only presented for D. acutus envenomation cases (Both 53.3% vs. 0.0%, P = 0.003). As to laboratory features, lower platelet counts (20.0 × 103/μL [interquartile range, 14–66 × 103/μL] vs. 149.0 × 103/μL [102.3–274.3 × 103/μL], P = 0.001), lower D-dimer level (1423.4 μg/L [713.4–4212.3 μg/L] vs. 12,500.0 μg/L [2351.4–200,000 μg/L], P = 0.008), higher proportion of patients with moderate-to-severe thrombocytopenia (platelet count < 100 × 103/μL) (80% vs. 16.7%, odds ratio (OR) = 20.0, 95% CI, 2.77–144.31; P = 0.002), and lower proportion of patients with extremely high D-dimer (> 5000 ng/mL) (16.7% vs. 66.7%, adjusted OR = 0.1 (95% CI, 0.01–0.69; P = 0.036) were found among cases of D. acutus envenomation compared to D. siamensis envenomation. The combination of hemorrhagic bulla, thrombocytopenia, and a lack of extremely high D-dimer had good discriminatory power (area under the curve (AUC) = 0.965; 95% CI, 0.904–1.00) for distinguishing D. acutus from D. siamensis envenomation.ConclusionsThe presentation of moderate to severe thrombocytopenia (platelet count < 100 × 103/μL) and hemorrhagic bulla formation may indicate D. acutus envenomation. However, the envenomed patient with extremely high D-dimer levels may indicate a D. siamensis envenomation. These findings may help diagnose and select the right antivenom in patients with unknown snakebites who present significant coagulopathy.

Morphometric characteristics and seasonal proximity to water of the Cypriot blunt-nosed viper Macrovipera lebetina lebetina (Linnaeus, 1758)

BackgroundThe blunt-nosed viper Macrovipera lebetina (Linnaeus, 1758) is a medically important snake species in the Middle East. Its nominate subspecies Macrovipera l. lebetina is confined to Cyprus, where it is the only dangerously venomous snake species and heavily pursued. Despite the viper’s large size, data on its body mass and sex-specific morphological differences are scarce. It is commonly believed that M. l. lebetina prefers freshwater proximity during summer. Hence, we aimed at investigating M. l. lebetina sex-specific morphological differences and its possible attraction to freshwater bodies in late summer.MethodsMorphometric characteristics, proximity to water and conservation status of M. l. lebetina were investigated in Paphos district (Cyprus) in 2014, 2015 and 2017. Vipers were caught in different habitats, examined morphologically for metric and meristic characters, and released back into their habitat. Additionally, local people were interviewed about the conservation situation of the species.ResultsOf 38 recorded blunt-nosed vipers, morphological characteristics were collected from 34 (10 adult males, 16 adult females, eight unsexed juveniles). Rounded total length (ToL) ranged from 23.5 cm to 133.0 cm and weight between 10 g and 1456 g. Adult males significantly exceeded adult females in tail length (TaL), ToL and head length (HL). No significant sex-specific differences were found in snout-vent length (SVL), head width (HW), weight or body condition index (BCI), nor for the ratios TaL / SVL, TaL / ToL, HL / SVL or HL / HW. Adult females from late summer (2015) had a significantly lower mean BCI than those from spring (2014).Distances of blunt-nosed vipers to the nearest water bodies (natural and artificial, respectively) did not differ significantly between spring (2014) and late summer (2015). There was also no significant difference between the distances of vipers to natural and to artificial water bodies in spring (and late summer).ConclusionsAdult male blunt-nosed vipers exceed adult females in TaL, ToL and HL. Adult females are likely in a more vulnerable body condition in late summer than in spring. Periodic drying out of freshwater bodies in summer probably does not affect the species’ occurrence. Educational workshops and habitat conservation are recommended for reducing human-viper conflict.

Purification and characterization of cysteine-rich secretory proteins (SVCRIPSS) from 5 different species of North America venomous snakes

Purification and characterization of cysteine-rich secretory proteins (SVCRIPSS) from 5 different species of North America venomous snakes

Vulnerability to snakebite envenoming: a global mapping of hotspots

SummaryBackgroundSnakebite envenoming is a frequently overlooked cause of mortality and morbidity. Data for snake ecology and existing snakebite interventions are scarce, limiting accurate burden estimation initiatives. Low global awareness stunts new interventions, adequate health resources, and available health care. Therefore, we aimed to synthesise currently available data to identify the most vulnerable populations at risk of snakebite, and where additional data to manage this global problem are needed.MethodsWe assembled a list of snake species using WHO guidelines. Where relevant, we obtained expert opinion range (EOR) maps from WHO or the Clinical Toxinology Resources. We also obtained occurrence data for each snake species from a variety of websites, such as VertNet and iNaturalist, using the spocc R package (version 0.7.0). We removed duplicate occurrence data and categorised snakes into three groups: group A (no available EOR map or species occurrence records), group B (EOR map but <5 species occurrence records), and group C (EOR map and ≥5 species occurrence records). For group C species, we did a multivariate environmental similarity analysis using the 2008 WHO EOR maps and newly available evidence. Using these data and the EOR maps, we produced contemporary range maps for medically important venomous snake species at a 5 × 5 km resolution. We subsequently triangulated these data with three health system metrics (antivenom availability, accessibility to urban centres, and the Healthcare Access and Quality [HAQ] Index) to identify the populations most vulnerable to snakebite morbidity and mortality.FindingsWe provide a map showing the ranges of 278 snake species globally. Although about 6·85 billion people worldwide live within range of areas inhabited by snakes, about 146·70 million live within remote areas lacking quality health-care provisioning. Comparing opposite ends of the HAQ Index, 272·91 million individuals (65·25%) of the population within the lowest decile are at risk of exposure to any snake for which no effective therapy exists compared with 519·46 million individuals (27·79%) within the highest HAQ Index decile, showing a disproportionate coverage in reported antivenom availability. Antivenoms were available for 119 (43%) of 278 snake species evaluated by WHO, while globally 750·19 million (10·95%) of those living within snake ranges live more than 1 h from population centres. In total, we identify about 92·66 million people living within these vulnerable geographies, including many sub-Saharan countries, Indonesia, and other parts of southeast Asia.InterpretationIdentifying exact populations vulnerable to the most severe outcomes of snakebite envenoming at a subnational level is important for prioritising new data collection and collation, reinforcing envenoming treatment, existing health-care systems, and deploying currently available and future interventions. These maps can guide future research efforts on snakebite envenoming from both ecological and public health perspectives and better target future estimates of the burden of this neglected tropical disease.FundingBill & Melinda Gates Foundation.

Mass spectrometry-assisted venom profiling of Hypnale hypnale found in the Western Ghats of India incorporating de novo sequencing approaches

Hypnale hypnale (hump-nosed pit viper) is considered to be one among the medically important venomous snake species of India and Sri Lanka. In the present study, venom proteome profiling of a single Hypnale hypnale from Western Ghats of India was achieved using SDS-PAGE based protein separation followed by LC-MS/MS analysis. The identities of the proteins that were not established using the Mascot search were determined through de novo sequencing tools such as Novor followed by MS-BLAST based sequence similarity search algorithm and PEAKS proteomics software. The combined proteomics analysis revealed a total of 37 proteins belonging to nine different snake venom families, in which 7 proteins were exclusively identified through de novo strategies. The enzymatic and non-enzymatic venom protein families identified include serine proteases, metalloproteases, phospholipase A2, thrombin-like enzymes, phospholipase B, C-type lectins/snaclecs, disintegrins, cysteine rich secretory proteins and nerve growth factor. Among these, disintegrins, nerve growth factor, phospholipase B and cysteine rich secretory protein families were identified for the first time in HPV venom. This could possibly explain the regiospecific venom variation seen across snake species. Taken together, the venom proteome profiling on Indian Hypnale hypnale venom correlates with the clinical manifestations often seen in the envenomed victims.

Proteomic characterization of six Taiwanese snake venoms: Identification of species-specific proteins and development of a SISCAPA-MRM assay for cobra venom factors

Deinagkistrodon acutus, Trimeresurus stejnegeri, Protobothrops mucrosquamatus, Daboia russelii siamensis, Bungarus multicinctus and Naja atra are the six medically important venomous snake species in Taiwan. In this study, we characterized and compared their venom protein profiles using proteomic approaches. The major snake venom proteins were identified by GeLC-MS/MS and the total venom proteome was characterized by in-solution digestion coupled with LC-MS/MS. A total of 27–52 proteins, categorized into 23 protein families, were identified in each snake's venom. The major venom components found in Viperidae species (D. acutus, T. stejnegeri, P. mucrosquamatus and D. russelii) were C-type lectin, snake venom serine proteinase, venom metalloproteinase and phospholipase A2, whereas three-finger toxin and phospholipase A2 were the major components detected in the venom of Elapidae snakes (B. multicinctus and N. atra). This study also provided the first demonstration of some low-abundance proteins in these six snake venoms, including 5′-nucleotidase, glutaminyl-peptide cyclotransferase and phosphodiesterase, among others. Furthermore, we found that cobra venom factor (CVF) is a cobra-specific protein. We produced anti-peptide antibodies against CVF and used it to develop a highly sensitive SISCAPA-MRM assay for quantifying CVF. The limit of detection and lower limit of quantification were 3.2 and 9.6 attomoles, respectively. This assay was used to precisely quantify CVF in 1 μg crude venom proteins from three Naja species and king cobra. The amount of CVF varied from 0.9 to 54.36 femtomoles (equivalent to 0.16–10.03 mg/g of venom protein). Biological significanceThere are six medically significant venomous snakes in Taiwan. The venoms of the four Viperidae species (Deinagkistrodon acutus, Trimeresurus stejnegeri, Protobothrops mucrosquamatus and Daboia russelii siamensis) cause local tissue swelling; this symptom is also seen in N. atra envenomation in humans, potentially complicating the differential diagnosis of envenomation by N. atra and Viperidae species. Thus, characterization of the venom proteomes of the six Taiwanese snakes, including the relative abundance of the major components and species-specific protein(s) in each venom type, could be useful for future venom research, including the development of new assay(s) for detecting snake species-specific venom protein(s) and new type(s) of antivenom.

Snakebites Treated in North Carolina Emergency Departments, October 2013–September 2015

North Carolina (NC) is home to more than 30 species of indigenous venomous and nonvenomous snakes. Snakebites can cause debilitating and potentially fatal injuries. However, there is a lack of current information available describing the incidence of snakebites in NC. Therefore, we performed this study of snakebites treated in NC emergency departments (EDs) using the statewide syndromic surveillance system, the North Carolina Disease Event Tracking and Epidemiologic Collection Tool (NC DETECT). This was a descriptive epidemiologic study characterizing NC ED visits collected by NC DETECT between October 1, 2013 and September 30, 2015 with an assigned International Classification of Diseases, 9th Revision, Clinical Modification code or keyword indicating a snakebite. Over the 2-year period, the absolute count of snakebite-related ED visits was 2080 visits with an incidence rate of 10.4 visits per 100 000 person-years (95% confidence interval: 10.0-10.9). The frequency of snakebite was highest during the summer months and evening hours. Men had higher incidence rates of snakebite-related ED visits than women, and residents of the Coastal Plain geographic region of NC had higher incidence rates than persons in other regions. The current study indicated that snakebites are common injuries treated at NC EDs, with a strong seasonal and geographic component. Additional research is needed to further characterize the circumstances associated with snakebites for the development of preventive measures and public health education.

Snake bite -A review

Snake bite is a common but under-reported medical emergency accounting for 0.5% of all deaths with greater risk of fatal envenomation in children. In India, four species of venomous snake are most common but better classification of medically significant species is warranted. Snake venom is a mixture of peptides with enzymatic & toxic properties which mediate activation of cytokine cascades along with organ specific toxicities, manifesting into local and systemic symptoms. The syndromic approach of attributing a constellation of signs & symptoms to a particular family of venomous snake has clinical acceptance but overlaps exist. Management of snake bite victim starts with a first aid measures of reassurance, immobilization and quick transfer to hospital. Measures such as application of tourniquet, incision and suction are harmful & should not be done. On arrival at hospital, triage and stabilization of Airway, Breathing & Circulation (ABC) is done followed by a focused assessment to ascertain the severity of envenomation. Antivenom treatment is the mainstay of snake bite management. ASV should be started only when specific indications such as signs of neurotoxicity, coagulopathy, hypotension, hematuria are present. Indiscriminate use of ASV is strongly condemned. Currently 8-10 vials of ASV as initial dose with a maximum of 25 vials is recommended. There is no role of test dose of ASV. Measures to treat any ASV induced anaphylaxis should be ready prior to start of ASV treatment. Supportive treatment is as important in determining the final outcome of envenomation as ASV

Spatial ecology of a small arboreal ambush predator, Trimeresurus macrops Kramer, 1977, in Northeast Thailand

Abstract The Big-Eyed Green Pit Viper (Trimeresurus macrops; Kramer, 1977) is a venomous snake species endemic to Southeast Asia. Although we have some knowledge of the systematics and toxicology of T. macrops, little is known about the spatial ecology of this species. From May 2013 to February 2014, we used radio-telemetry to determine home-range sizes of 13 adult female T. macrops inhabiting the Sakaerat Biosphere Reserve in Northeast Thailand. We found that individual home ranges for T. macrops averaged 0.175 ha, with activity areas ranging from 0.112-0.303 ha and core areas ranging from 0.023-0.052 ha. There was little overlap between conspecific tracked females, especially for the most used areas of their home ranges. We find that T. macrops ambushes more in higher humidity and expresses very little diurnal activity. They use the groundstory for ambushing, then retreat over small distances to higher refuge during the day. Future studies should focus on prey abundance, habitat selection, and survival rates.

The effects of Macrovipera lebetina venom on mice

The aim of this study is to reveal the effects Macrovipera lebetina venom – one of the two most venomous (the other one is Montivipera xanthina) snake species in Turkey – histopathologically on liver, kidney and heart tissues of mice. The venom was injected into the tail veins of male mice at three different doses (0.5, 1 and 1.5 mg kg−1). The animals were dissected 2 h after the injection, and their liver, kidney and heart tissues were examined histologically by light microscopy. Results of histopathological examination showed, degeneration, mononuclear cell infiltration, vacuoles around the nuclei and sinuzoidal congestion in livers; cellular degeneration, deformation in the structure of the glomerulus, intertubular congestion and mononuclear cell infiltration in the kidneys; and vacuoles around the nucleus, necrosis, congestion and mononuclear cell infiltration in heart tissues.

Computational Studies of Snake Venom Toxins.

Most snake venom toxins are proteins, and participate to envenomation through a diverse array of bioactivities, such as bleeding, inflammation, and pain, cytotoxic, cardiotoxic or neurotoxic effects. The venom of a single snake species contains hundreds of toxins, and the venoms of the 725 species of venomous snakes represent a large pool of potentially bioactive proteins. Despite considerable discovery efforts, most of the snake venom toxins are still uncharacterized. Modern bioinformatics tools have been recently developed to mine snake venoms, helping focus experimental research on the most potentially interesting toxins. Some computational techniques predict toxin molecular targets, and the binding mode to these targets. This review gives an overview of current knowledge on the ~2200 sequences, and more than 400 three-dimensional structures of snake toxins deposited in public repositories, as well as of molecular modeling studies of the interaction between these toxins and their molecular targets. We also describe how modern bioinformatics have been used to study the snake venom protein phospholipase A2, the small basic myotoxin Crotamine, and the three-finger peptide Mambalgin.

Is coagulopathy a common consequence of a Vipera berus bite? A retrospective single centre study.

The Vipera berus (common viper) is the only species of venomous snake found in Poland. The aim of this study was to determine to what extent coagulopathy has occurred as a post-bite complication. The medical records of 138 adult patients in the Sucha Beskidzka hospital with the diagnosis of snake bite between 2001-2014 were retrospectively analyzed. Demographic, clinical and laboratory data were collected. Antivenom was given to all patients, except one, with snake bites, but laboratory test were taken prior to administration of antivenom. Obtained results were compared to those of the control group, composed of 176 adults hospitalized in the same period for an elective laparoscopic cholecystectomy. The mean platelet count in the study group was 239.94×109/L (SD=56.56) and 248.77×109/L (SD=57.82) in the control group. In 98% of the study group and 100% of the control group the platelet value (PLT) fell within laboratory norms (130 to 420×109/L), 2 patients after snake bites had a PLT lower than normal, and thrombocytosis was not observed in either group. A reference range of 0.85-1.3 INR below normal was found in 2 patients in the study group and 15 from the control group, while values above the normal range were found in 8 patients (6.7%) from the snake bite group and no patients from the control group. In the study group, the INR ranged from 0.78-1.43 with a mean of 1.046 (SD=0.14), while in the control group the range was from 0.79-1.28, with a mean of 0.95 (SD=0.08). There was a significant difference in INR between the study and control groups (p<0.0001). The bite from V. berus does not result in severe thrombocytopenia, with only a moderate increase in INR values observed in about 7% of patients.

Diversity and Habitat Preferences of Herpetofauna at Kalabagh Game Reserve, District Mianwali, Punjab, Pakistan

During present survey extending from July, 2015 through March, 2016 six amphibian species representing 6 genera and 3 families and 24 reptilian species representing 23 genera and 13 families, were recorded from Kalabagh Game Reserve, district Mianwali, Pakistan. Among the five capture techniques, i.e., hand picking, use of snake sticks, forceps, drag nets and noose traps, hand picking appeared the most effective one. The captured amphibian species included Bufo stomaticus, Microhyla ornata, Euphlyctis cyanophlyctis, Fejervarya limnocharis, Hoplobatrachus tigerinus, and Sphaeroteca breviceps. Single turtle species, Lissemys punctata andersoni was recorded from the study area. Among lizard species, Calotes versicolor, Laudakia nupta, Trapelus agilis pakistanensis, Eublepharis macularis, Cyrtopodion montiumsalsorum, Hemidactylus brookii, Hemidactylus flaviviridis, Ablepharus grayanus, Eutropis macularia, Lygosoma punctatum, Novoeumeces indothalensis, Uromastyx hardwickii, and Varanus bengalensis were recorded. Similarly, snake species included Leptotyphlops macrorhynchus, Ramphotyphlops braminus, Eryx johnii, Amphiesma stolatum, Platyceps rhodorachis, Lycodon aulicus, Ptyas mucosus, Bungarus caeruleus, Naja oxiana, and Echis carinatus. The venomous snake species of the study area included Naja oxiana, Bungarus caeruleus, and Echis carinatus.