Treatment Of Snakebite Research Articles

Protective effects of novel diazepinone derivatives in snake venom induced sterile inflammation in experimental animals

Snake envenomation leads to the formation of damage–associated molecular patterns (DAMPs), which are mediated by endogenous intracellular molecules. These are recognized by pattern-recognition receptors (PRRs) and can induce sterile inflammation. AimsIn the present study, we aim at understanding the mechanisms involved in DAMPs induced sterile inflammation to unravel the novel therapeutic strategies for treating snake bites. The potential of benzodiazepinone derivatives to act against snake venom induced inflammation has been explored in the present investigation. Main methodsThree compounds VA 17, VA 43 and PA 03 were taken from our library of synthetic compounds. Oxidative stress markers such as lipid peroxidation, superoxide and nitric oxide were measured along with the analysis of DAMPs (IL6, HMGB1, vWF, S100b and HSP70). These compounds have been docked using molecular docking against the snake venom PLA2 structure (PDB code: 1OXL). Key findingsThe compounds have been found to effectively neutralize viper and cobra venoms induced lethal activity both ex vivo and in vivo. The compounds have also neutralized the viper venom induced hemorrhagic, coagulant, anticoagulant reactions as well as inflammation. The fold of protection have always been found to be higher in case of ex vivo than in in vivo. These compounds have neutralized the venom induced DAMPs as exhibited by IL6, HMGB1, vWF, S100b and HSP70. The fold of neutralization is found to be higher in VA 43. SignificanceThe identified compounds could be used as potential candidates for developing treatment of snakebites in areas where antiserums are not yet available.

Larvicidal Activity of Leucas Aspera Against the Larvae of Anopheles, Aedes Aegypti and Culex Quinquefasciatus

Abstract: Insect-transmitted diseases cause high morbidity however they additionally embody deadly diseases that cause high mortality rates among infected individuals.Many two-winged insects species such as Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus act as vectors. Ae. aegypti, acts as a vector for yellow fever, dengue and chikungunya. Aedes aegypti is cosmopolitan within the tropical and semitropical zones. Anopheles stephensi is the primary vector of malaria in India and other west Asian countries. C. quinquefasciatus is a vector of lymphatic filariasis which is one of the widely distributed tropical diseases with around 120 million people infected worldwide. The mosquitocidal activity was analyzed by aqueous, ethanol and methanol extract of Leucas aspera plant extract against the malaria, yellow fever and dengue causing Ae.aegypti, An.stephensi. Leucas aspera is well known to possess various pharmacological activities like antifungal, antimicrobial and cytotoxic activities. Leaves of this plant are applied to treat snake bites. The result was supported by probit analyses was carried out for twenty fourth and forty eighth hour methyl alcohol extracts of L. aspera. The data showed profound larvicidal activity with methyl alcohol extract compared with the other extracts. Keywords: Vector control, L. aspera, Ae.a.egypti, An. stephensi, Culex quinquefasciatus.

Ethnobotanical survey of medicinal plants against ophidian envenomations in the Bonginda/Bikoro Group in DR Congo

Snakebite envenomation is a neglected tropical disease causing enormous suffering, disability and premature death on all continents. This study aims to inventory the plants involved in the management of snakebite cases in rural areas. A survey was conducted in the Bonginda groupement, Bonginda Territory, Equateur Province in DR Congo during the period from January 2019 to December 2020. Data was collected by the standardized interview method with a focus on individual interview technique. To this end, a questionnaire was administered to traditional healers reputed in the treatment of snakebites. It shows that 17 species grouped into 14 families are used to treat cases of ophidian envenomation in Bonginda. The Fabaceae and Rubiaceae families each have 3 species. The others are monospecific. Among the organs used, leaves predominate with a score of 44.4%. They are followed by the roots (18.5%). Regarding the mode of preparation, the decoction prevails with 27.3%. It is followed by grinding (13.6%). As for the route of administration, local application (28.6%) is ahead of the oral route (23.8%) and purgation (14.3%). Constituents based on medicinal plants are recognized as local heritage.

Phytochemical analysis and preclinical toxicological, antioxidant, and anti-inflammatory evaluation of hydroethanol extract from the roots of Harpalyce brasiliana Benth (Leguminosae)

Ethnopharmacological relevanceHarpalyce brasiliana Benth (Leguminosae) is a shrub endemic to Brazil, popularly known as “snake's root.” This species is used in folk medicine for the treatment of inflammation and snakebites. However, up to now there is no scientific research to justify its popular use. The study aimed to characterize the phytochemical profile of the hydroethanol extract from the roots of H. brasiliana (Hb), to evaluate its antioxidant and anti-inflammatory potential, as well as to investigate its cytotoxicity and acute toxicity. Materials and methodsThe extract was obtained by maceration method using a solution of ethanol:water (70: 30, v/v). The phytochemical profile was obtained by liquid chromatography coupled to mass spectrometry. The cytotoxicity of extract (31–2000 μg/mL) was evaluated in vitro, by the 3-methyl-[4-5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) method using murine macrophage and fibroblast cell lines (RAW 247.6 and 3T3, respectively) and by the hemolytic assay. For the in vivo acute toxicity, the extract (2000 mg/kg) was administered and after 14 days the weight (body and organs) and hematological and biochemical parameters were analyzed. Chemical free radical scavenging effect of the extract (125–2000 μg/mL) was investigated through diphenylpicryl hydrazine reduction, total antioxidant capacity, reducing power, hydroxyl radical scavenging, and iron and copper chelating assays. In vitro anti-inflammatory effect of the extract (125, 500, and 2000 μg/mL) was demonstrated through of nitric oxide (NO) analyzed in lipopolysaccharides stimulated RAW 264.7 cells. In vivo anti-inflammatory activities were evaluated in carrageenan-induced paw edema and zymosan-air-pouch models, with gavage administration (post-treatment) of extract at 100, 200, and 400 mg/kg. For the first animal model, the anti-edematogenic activity and myeloperoxidase (MPO) levels were investigated, while in the zymosan-air-pouch model the leukocyte number, MPO, total protein and pro-inflammatory cytokine (IL-1β, IL-6, and TNF-α) levels were quantified. In addition, the oxidative parameters such as malondialdehyde (MDA) and reduced glutathione (GSH) were determined. ResultsThe phytochemical profile revealed the presence of 20 compounds, mainly prenylated and geranylated pterocarpans. The extract demonstrated no cytotoxicity in erythrocytes, macrophages and fibroblasts cells at the tested concentrations, as well as no sign of toxicity and mortality or significant alterations on the hematological and biochemical parameters in the acute toxicity model. The extract was also able to neutralize chemical free radicals, with copper and iron chelating effect. For the NO dosage, the extract evidenced the reduction of expression of NO after the administration of the extract (500 and 2000 μg/mL). The edematogenic model revealed a decrease in paw edema and MPO level, while the zymosan-air-pouch model evidenced a reduction of leukocyte number (especially of polymorphornuclears), MPO production, and total protein and cytokine levels, and demonstrated the antioxidant effect through a decrease in MDA and increase in GSH parameters. ConclusionThis approach demonstrates for the first time that Hb is not cytotoxic, has low acute toxicity, and possesses antioxidant and anti-inflammatory properties in preclinical analyses, corroborating its popular use.

A comparative study of endogenous phospholipase A2 inhibitors in the serum of Brazilian pit vipers

This work compared the presence of phospholipase A2 inhibitors (PLIs) in the serum of 19 snake species maintained at Instituto Butantan to better understand the mechanisms of venom resistance in snakes and improve the treatment of snakebite. PLI was isolated from blood of 19 snake species by one-step chromatography and identified in all samples, besides its identity was confirmed through the interaction with both phospholipase A2 and anti-γPLI. These findings highlight the diversity of snake serum PLIs and emphasize the importance of structure-function studies.

Snakebite Treatment in Tanzania: Identifying Gaps in Community Practices and Hospital Resources.

Snakebite envenoming causes more than 140,000 deaths annually and at least triple this number of disabilities. The World Health Organization classified snakebite as a Neglected Tropical Disease in 2017 and developed a strategy to halve death and disability from snakebite by 2030. To achieve this goal, snakebite victims need to receive safe and effective treatment. This descriptive, cross-sectional study surveyed student health professionals (N = 312) in Dar es Salaam, Tanzania, and was designed to identify major gaps in community practices and hospital resources for snakebite treatment. Participants reported using traditional community practices (44%, 95% confidence interval (CI) = 39–50%), allopathic practices (7%, 95% CI = 5–11%), or a combination of both (49%, 95% CI = 43–54%) to treat snakebite. Harmful practices included tight arterial tourniquets (46%, 95% CI = 41–52%) and wound incisions (15%, 95% CI = 11–19%). Many participants (35%, 95% CI = 29–40%) also turned to traditional healers. Students who treated snakebite injuries within the last 5 years (N = 69) also reported their general experiences with snakebite in hospitals. Hospitals often lacked essential resources to treat snakebite victims, and 44% (95% CI = 30–59%) of snakebite victims arrived at a hospital only three or more hours after the bite. A significant percentage of snakebite victims experienced lasting damage (32%, 95% CI = 20–47%) or death (14%, 95% CI = 7–25%). Snakebite outcomes could likely be improved if hospitals were universally and consistently equipped with the essential resources to treat snakebite victims, such as antivenoms. Educational interventions aimed at communities should focus on discouraging tourniquet use and tampering with the wound. Collaboration between the allopathic and traditional health system could further boost snakebite outcomes because traditional healers are often the first health workers to see snakebite victims.

Tandem high resolution mass spectrometry based phytochemical composition of Sauromatum guttatum tubers and its enzyme inhibitory potential with molecular docking

Sauromatum guttatum has been traditionally used in the treatment of snakebite and tumors in India, Pakistan, and China. However, it lacks detailed phytochemical composition like other members of the family Araceae. Therefore, the aim of the present study was to investigate the phytochemical composition of crude methanolic extract and subsequent fractions from S. guttatum tubers and to determine their enzyme inhibitory potentials. The phytochemical profile was studied through tandem high-resolution mass-based phytochemical analysis and Global Natural Product Social (GNPS) molecular networking. Similarly, crude extract and fractions were also investigated for enzyme inhibitory activity against urease and α-glucosidase. Twenty-six compounds were dereplicated belonging to flavone C-glycosides, flavone O-glycosides, phenolic acids, phenolic acid glycosides, and iridoid glycosides. The n-butanol fraction was particularly found rich in flavone di-C-glycosides including schaftoside, isoschaftoside, neoschaftoside, and vicenin-2. The n-butanol fraction exhibited the highest in vitro inhibition against urease and α-glucosidase with IC50 values of 113.7 µg/mL and 155.3 µg/mL, respectively. The results of enzyme inhibition potential were also supported by in silico molecular docking studies against the above-mentioned enzymes. This is the first report on the detailed phytochemical profile of S. guttatum tubers, and these results will contribute to the chemosystematic knowledge of the Araceae family. The results of this study also suggest that S. guttatum may find possible applications in the treatment of gastrointestinal disorders and diabetes.

Anti-Neurotoxins from Micrurus mipartitus in the Development of Coral Snake Antivenoms.

In Colombia, the genus Micrurus includes 30 species, of which M. mipartitus and M. dumerilii are the most widely distributed. Micrurus causes less than 3% of the approximately 5000 cases of snakebite per year. The elapid envenomation caused by the snakes from the Micrurus genus, are characterized by the severity of their clinical manifestations, due to the venom neurotoxic components such as three-finger toxins (3FTx) and phospholipases (PLA2). The treatment for snakebites is the administration of specific antivenoms, however, some of them have limitations in their neutralizing ability. A strategy proposed to improve antivenoms is to produce antibodies against the main components of the venom. The aim of this work was to produce an antivenom, using an immunization protocol including the main 3FTx and PLA2 responsible for M. mipartitus lethality. The antibody titers were determined by ELISA in rabbits’ serum. The immunized animals elicited a response against toxins and whole venom. The Immunoglobulin G (IgGs) obtained were able to neutralize the lethal effect of their homologous toxins. A combination of antivenom from M. mipartitus with antitoxins improved their neutralizing ability. In the same way, a mixture of anti 3FTx and PLA2 protected the mice from a 1.5 median lethal dose (LD50) of M. mipartitus venom. The results showed that this might be a way to improve antibody titers specificity against the relevant toxins in M. mipartitus venom and indicated that there is a possibility to develop and use recombinant 3FTx and PLA2 toxins as immunogens to produce antivenoms. Additionally, this represents an alternative to reduce the amount of venom used in anti-coral antivenom production.

Isolation and characterization of anti-inflammatory and anti-proliferative compound, for B-cell Non-Hodgkin lymphoma, from Nyctanthes arbor-tristis Linn.

Ethnopharmacological relevanceNyctanthes arbor-tristis Linn. is native to Indo-Pak sub-continent and has high medicinal values in Ayureda. This plant has been used traditionally for the treatment of sciatica, rheumatism, chronic fever, diabetes, snakebite, dysentery, cachexia and cancer. Studies have shown many pharmacological properties such as anti-cancer efficacy against Dalton's ascetic lymphoma, cytotoxicity against T-cell leukemia, anti-inflammatory, anti-diabetic and anti-oxidant effects. Aim of the studyAim of the study was to explore the anti-inflammatory and anti-proliferative potential of N. arbor-tristis. Material and methodsEthanol extract of fresh and uncrushed aerial parts of N. arbor-tristis was used in the present study. A new compound nyctanthesin A was isolated following a bioactivity-guided fractionation and chromatographic separations. Its chemical structure was elucidated through spectral studies including 1D, 2D-NMR experiments and HREIMS. The intracellular reactive oxygen species (ROS) and nitric oxide (NO) generation from phagocytes were detected by chemiluminescence technique and Griess method, respectively. TNF-α and TGF-β production was quantified by ELISA. Anti-lymphoma and cytotoxic activities were assessed by alamar blue and MTT assays, respectively. The transcription and protein expression level of Bcl-2, COX-2, p38 MAPK, PDL-1, NF-κB, c-Myc and PNF-κB was performed by qRT-PCR and protein blot assays, respectively. ResultsPetroleum ether insoluble fraction of the ethanol extract of fresh and uncrushed aerial parts of N. arbor-tristis revealed anti-inflammatory potential by inhibiting ROS. A previously undescribed compound nyctanthesin A was isolated from this fraction and characterized by UV, IR, NMR and HREIMS. It showed significant anti-inflammatory property by inhibiting ROS, NO and TNF-α production. The strong anti-proliferative effects on B- cell lymphoma cells, DOHH2 and Raji, revealed its anti-lymphoma potential along with non-toxic profile against BJ and NIH-3T3 fibroblast cells of normal origin. The qRT-PCR results showed marked inhibition of Bcl-2, COX-2, p38 MAPK, PDL-1, c-Myc, NF-κB, and PNF-κB at transcription level in DOHH2 cells with comparatively lesser but significant effects in Raji cells, where the expression of Bcl-2 gene was not affected. The protein expression of PNF-κB in DOHH2 cells was inhibited by 66% (P < 0.05) and COX-2 in both cell lines was inhibited by 50% (P < 0.05) at 60 μg/mL. A moderate non-significant inhibition of TGF-β (∼20%) was observed in both cell lines at 100 μg/mL ConclusionsScientific evidences reported here validate the anti-inflammatory and anti-cancer potential of the plant.

A Review on the Morphology, Nutritional Value, Traditional Uses, Phytochemistry, and Biological Activities of Pycnarrhena cauliflora and Its Synonyms

The leaves of Pycnarrhena cauliflora, locally known as ‘pokok Ajinomoto’, are consumed as flavour enhancers by East Malaysian communities. The plant also plays a significant role in traditional medicine especially to treat snake bites, eye irritation, headache, and seizures. An extensive literature search on the species revealed that besides taxonomic description as well as toxicity against HeLa human cervical and breast cancer cell lines, there is no report on the plant chemistry. The species has nevertheless been classified as the synonym of several other species which are Antitaxis cauliflora Miers., Pycnarrhena longifolia, (Decne. ex Miq.), Antitaxis longifolius (Decne. ex Miq.) Mier., and Gabila longifolia (Decne. ex Miq, B). Among these synonyms, the plant is closely related to the species P. longifolia. Thus, a complete review of P. cauliflora and its synonyms is important to revise and evaluate its potential for further studies and commercialisation. The information on the species was collected from scientific journals, books, and reports searched through available databases such as Google Scholar, PubMed, Directory of Open Access Journals, Science Direct, Bioline International and Reaxys. This review provides an insight on the morphology, nutritional value, traditional uses, phytochemistry, and biological activities of P. cauliflora and its synonyms published between 1981 to 2021 in scientific journals, books, and reports.

Combined proteomic strategies for in-depth venomic analysis of the beaked sea snake (Hydrophis schistosus) from Songkhla Lake, Thailand

This study focuses on comprehensive characterization of the venom proteome of the beaked sea snake (Hydrophis schistosus) from Songkhla Lake, Thailand. H. schistosus can be considered as the deadliest sea snake commonly found in the Pacific and Indian oceans. Their envenomation causes muscular paralysis and rhabdomyolysis. To develop effective treatment for this snakebite, it is necessary to understand the detailed venom composition. In this study, multiple mass spectrometry-based approaches were employed. Bottom-up proteomics revealed that tryptic digestion in-solution provided a higher number of toxin proteins identified and a larger sequence coverage, compared to in-gel digestion. In addition, a venom gland transcriptome-derived database was constructed and used as a reference, which 43 known and novel toxin proteins were identified using this database and the UniProtKB. Three-finger toxin and phospholipase A2 were shown to be top two most abundant protein families. Minor compositions included other toxin families and a number of non-toxin proteins. Moreover, a hybrid de novo sequencing was performed to enhance identification of the small proteins/peptides. Using non-digested samples, there were 46 predicted toxin peptides. The finding from this study could lead to a better understanding in pathological effects of the snakebite and the future development of effective antivenoms. SignificanceThis study provides a better understanding of the venom proteome composition of the beaked sea snake (H. schistosus) found in the Gulf of Thailand, using a combination of different sample preparation techniques, Serpentes protein database searching, transcriptome-derived protein database searching, and a hybrid de novo peptide sequencing strategy. It revealed 13 toxin protein families and novel proteins in the beaked sea snake venom including new species of phospholipase A2s (PLA2s) and three-finger toxins (3FTxs). It could serve as a basis for the development of snakebite treatments and for the discovery of novel pharmaceutical drugs from the toxin peptides.

Insights on the inhibition properties of Jatromollistatin (a cyclic heptapeptide) against Crotalus adamanteus metalloendopeptidase using molecular docking analysis.

Jatropha mollissima is endemic to Brazil and is used for traditional medicinal purposes, including the treatment of snakebite. In this study, latex obtained from this plant was fractioned using reversed-phase chromatography, and the fractions were then screened for peptides. A 755 g/mol peptide was obtained, and MS/MS analyses indicated it had a cyclic sequence (Pro-Leu-Gly-Val-Leu-Leu-Tyr). This peptide sequence was present in the Jatropha genome database, and an identity value of 90.71%, an E-value of 0.0, and a score of 883 with NO-associated protein 1/chloroplastic/mitochondria of Jatropha curcas were obtained from the NCBI nonredundant protein sequence (nr) database. Molecular docking analyses performed with the peptide against a metalloendopeptidase belonging to Crotalus adamanteus snake venom suggested the cyclic peptide establishes favorable interactions with the catalytic site of the enzyme. Therefore, it could inhibit enzyme catalysis. This belief was corroborated by the formation of 6 hydrogen bonds with the linear form of the peptide. Tighter complexation of the cyclic form (41 kcal/mol more energetic) revealed better spatial blocking. The linear form outperformed the cyclic form in complexing the required energy, recruiting more catalytic residues (6/2), and in establishing more hydrogen bonds (6/3). However, cyclic folding provided a more significant spatial block within the catalytic site. The set of results suggests that the cycle peptide, here called Jatromollistatin, which was previously described as jatrophidin and pohlianin A in two other species of Jatropha, is a promising candidate to inhibit venom proteases. This belief is corroborated by the topical use of the latex for initial treatment of snakebites.

The molecular basis of venom resistance in the non-venomous snake Sinonatrix annularis

Various snake species and snake predators have natural neutralization against snake toxins, which their antidotal abilities are commonly attributed to the intrinsic inhibitors produced by the liver, e.g., phospholipase A2 inhibitor (PLI) and metalloproteinase inhibitor (SVMPI). Sinonatrix annularis was found to possess broad-spectrum neutralization to different snake venoms in our lab. Although the anti-venom compound PLIγ has been previously characterized in our laboratory, the mechanism of resistance of S. annularis to snake venoms remains obscure. In this research, a venom affinity chromatography was constructed by immobilizing D. acutus venom to NHS-agarose beads and applied for antitoxins mining from S. annularis. The binding capacity of the venom column was validated using a self-prepared rabbit antivenom against D. acutus. Serum and liver homogenate of S. annularis were then applied to the column, the bound components were profiled using SDS-PAGE and mass spectrometry. PLIs, snake venom metalloproteins inhibitor (SVMPI), small serum protein (SSP), heat shock proteins, etc were identified. To identify their toxin targets in D. acutus venom, a reverse separation was conducted by coupling the fractionated S. annularis serum proteins to NHS-agarose beads. Fifteen toxins of five families were captured and identified as follows: PLA2s, metalloproteinases, cysteine-rich secretory proteins, snake venom serine proteinases, and C-type lectins. These discoveries increased our understanding of the capacity and mechanism of the natural neutralization of S. annularis to snake venom. These natural inhibitors are medically significant due to their powerful and broad antidotal activities, which may provide alternative and promising drug candidates for snakebite treatment.

The Preclinical Evaluation of a Second-Generation Antivenom for Treating Snake Envenoming in India

Snake envenoming afflicts the Indian subcontinent with the highest rates of mortality (47,000) and morbidity globally. The only effective treatment for snakebites is the administration of antivenom, which is produced by the hyperimmunisation of equines. Commercial Indian antivenoms, however, have been shown to exhibit a poor preclinical performance in neutralising venom, as a result of inter- and intrapopulation snake venom variation. Additionally, their poor dose effectiveness necessitates the administration of larger volumes of antivenom for treatment, leading to several harmful side effects in snakebite victims, including serum sickness and fatal anaphylaxis. In this study, we employed chromatographic purification to enhance the dose efficacy of commercial Indian antivenoms. The efficacy of this ‘second-generation’ antivenom was comparatively evaluated against six other marketed antivenoms using a number of in vitro and in vivo preclinical assays, which revealed its superior venom recognition capability. Enhanced purity also resulted in significant improvements in dose effectiveness, as the ‘second-generation’ antivenom exhibited a 3 to 4.5 times increased venom neutralisation potential. Furthermore, preclinical assays revealed the increased effectiveness of the ‘second-generation’ antivenom in countering morbid effects inflicted by the ‘big four’ Indian snakes. Thus, we demonstrate the role of simpler purification steps in significantly enhancing the effectiveness of snakebite therapy in regions that are most affected by snakebites.

Acute and subacute toxicity study of ethanolic extract of Calotropis procera (Aiton) Dryand flower in Swiss albino mice

BackgroundCalotropis procera is a large shrub which consists many medicinal properties, used in treatment of snake bite, sinus fistula, rheumatism, mumps, burn injuries, inflammation and jaundice traditionally. All the parts of Calotropis procera were utilized in the treatment of diseases out of which leaves and roots were investigated for its toxicity profile that showed dose dependent toxicity. Toxicity profile of flowers of Calotropis procera was not investigated in the previous studies. The aim of this study was to explore the acute and subacute toxicity of ethanolic extract of Calotropis procera flowers for the safe use of traditional medicine. MethodIn acute toxicity, a total of 20 female mice (Swiss albino), weighing between 23 and 32 g were randomly divided into four experimental groups: control, 300, 1000, and 2000 mg/kg groups with 5 mice each, and each received a single dose of extract at 300, 1000, or 2000 mg/kg, respectively. Animals were monitored for 14 days. In the subacute study, a total of 40 mice (23–32 g) were divided into 4 groups, each containing males and females. Group 1 (control group) received vehicle and groups 2, 3, and 4 received extract at doses of 300 mg/Kg, 1000 mg/Kg, 2000 mg/Kg of b.w., respectively, for 28 consecutive days. The study was conducted in compliance with the OECD guidelines 407 and 423. ResultsAcute toxicity study showed no mortality at the dose of 2000 mg/Kg. In subacute toxicity study, statistical analysis of hematological and biochemical parameters showed no significant differences compared to control group except marked increase in segmented neutrophils. Histopathological studies revealed no significant structural differences among the treated groups and in comparison to control group. ConclusionsIt was concluded that oral administration of doses of ethanolic extract of Calotropis procera flower, administered acutely, did not cause any mortality or notable changes at the dose of 2000 mg/Kg. Therefore, the approximate lethal dose (ALD) of in mice was higher than 2,000 mg/kg. In a 28-day subacute toxicity model, the extract did not cause any mortality, and no treatment-related changes were observed in body weight, organ weight, hematological and biochemical blood analysis, or histopathologic examinations at the extract dose of 2000 mg/Kg. These findings indicate that the no-observed-adverse-effect-level (NOAEL) of Calotropis procera flower ethanolic extract was greater than 2000 mg/kg/day.

Biodiversity of ethnomedicinal plants from the B’laan Tribe in Mount Matutum Protected Landscape, Southern Mindanao, Philippines

Abstract. Alinsug MV, Estandarte MHG, Somodio EMN, Sabarita MJJ, Deocaris CC. 2021. Biodiversity of ethnomedicinal plants from the B’laan Tribe in Mount Matutum Protected Landscape, Southern Mindanao, Philippines. Biodiversitas 23: 554-563. Mount Matutum Protected Landscape (MMPL) is one of the key biodiversity areas in Mindanao Island, Philippines, and is part of the ancestral domains of 18 indigenous peoples. The present study aims to establish a comprehensive inventory of the medicinal flora at MMPL according to the traditional healing practices of the B'laan, a tribal community whose cultural heritage is anchored to their interactions with nature and with a rich and understudied ethnomedicinal system. Focus group discussions and key informant interviews of the tribal healers from five tribal communities revealed 101 medicinal plants belonging to 33 families were identified. The plant species have therapeutic uses for 19 disease categories. Roots and leaves are the most utilized parts which are made into a decoction, the most common preparation for oral intake. Tawal-ulad (Lunasia amara Blanco), which is used as a treatment for snake bites, dengue, dysmenorrhea, constipation, rabies, and hypertension has the highest use value (UV). Informant Consensus Factor (ICF) analysis revealed that cholera, scabies, viral infections, warts, beriberi, diabetes, gastric ulcer are the most common diseases treated with medicinal plants by the B'laan tribal healers. The biodiversity of medicinal plants, although some remain unidentified at the species level, reflects the centuries-old ethnomedicinal knowledge of the B'laan Tribe. Our study is the first comprehensive documentation of the ethnomedicinal knowledge of the B'laan Tribe residing at MMPL.

Predictors of FabAV use in copperhead envenomation

Context Crotaline snake envenomation is a serious medical condition affecting thousands of Americans each year. Variation in the treatment of Crotaline snakebites exists among physicians in the United States. Management of copperhead snakebites is controversial with some experts advocating minimal intervention, rarely necessitating antivenom use and, even more rarely, surgical intervention. This study assessed the use of Crotaline Polyvalent Immune Fab antivenom (Ovine) (FabAV) and explored factors influencing the decision to prescribe antivenom for copperhead envenomation in patients in Northeastern Oklahoma. Methods A retrospective cohort study examining electronic medical records of patients with copperhead snakebites from July 1, 2014 to August 31, 2019. Data collected included: patient demographics, transfer information, snake species, bite site, progression of local tissue effects, additional clinical and lab results, patient comorbidities, and treatment strategy. Associations between patient variables and treatment were evaluated using the chi-square test of independence, median test, and logistic regression analysis. Associations were statistically significant if p < 0.05. Discussion Of the 130 patients bitten by a copperhead, a majority (75%) received FabAV. Symptoms of copperhead envenomation were mostly limited to the progression of tissue damage. Predictors of treatment with FabAV included progression of venom effects across major joints, younger age, comorbidities, and upper extremity bites. Conclusions Patients who have multiple comorbidities, upper extremity bites and progression of venom effects across major joints are more likely to be treated with FabAV. The high usage of FabAV at the study site underscores the need for continued work to optimize the use of antivenom for copperhead envenomations.

Screening and identification of differential metabolites in serum and urine of bamaxiang pigs bitten by &lt;i&gt;trimeresurus stejnegeri&lt;/i&gt; based on UPLC-Q-TOF/MS metabolomics technology

Trimeresurus stejnegeri is one of the top ten venomous snakes in China, and its bite causes acute and severe diseases. Elucidating the metabolic changes of the body caused by Trimeresurus stejnegeri bite will be beneficial to the diagnosis and treatment of snakebite. Thus, an animal pig model of Trimeresurus stejnegeri bite was established, and then the metabolites of serum and urine were subsequently screened and identified in both ESI+ and ESI- modes identified by ultra-performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS) methods. There are 9 differential metabolites in serum, including Oleic acid, Lithocholic acid, Deoxycholic acid, Hypoxanthine, etc. There are 11 differential metabolites in urine, including Dopamine, Thiocysteine, Arginine, Indoleacetaldehyde, etc. Serum enrichment pathway analysis showed that 5 metabolic pathways, including Tryptophanuria, Liver disease due to cystic fibrosis, Hartnup disease, Hyperbaric oxygen exposure and Biliary cirrhosis, the core metabolites in these pathways, including deoxycholic acid, lithocholic acid, tryptophan and hypoxanthine, changed significantly. Urine enrichment pathway analysis showed that 4 metabolic pathways, including Aromatic L-Amino Acid Decarboxylase, Vitiligo, Blue Diaper Syndrome and Hyperargininemia, the core metabolites in these pathways including dopamine, 5-hydroxyindole acetic acid and arginine. Taken together, the current study has successfully established an animal model of Trimeresurus stejnegeri bite, and identified the metabolic markers and metabolic pathways of Trimeresurus stejnegeri bite. These metabolites and pathways may have potential application value and provide a therapeutic basis for the treatment of Trimeresurus stejnegeri bite.

Next Generation Antivenoms and Their Neutralizing Efficacy against Snakebite Envenoming

Snake envenomation is a major neglected tropical disease and remains a serious threat in many countries at the present day. The impoverished rural populations are vulnerable to snakebite envenoming which eventually strengthens the cycle of poverty. The highly diversified components of snake venoms are primarily responsible for severe clinical manifestations in the victim. The only available treatment is the use of animal-derived broad-spectrum polyvalent antivenoms having a very low level of case-specific antibodies. A significant number of drawbacks of this antivenom are possessing a key challenge in snakebite treatments. The recent advancements into the toxin-specific monoclonal antibodies proved to be promising for future envenoming treatments, although its use is still in a naïve stage since it also holds some limitations in neutralizing the complex snake venoms. In the emerging fields of molecular biology along with transcriptomic and proteomic analysis protocols, the snake venom constituents have been well characterized and this holds another promising approach to using DNA immunization for next-generation therapeutics. This strategy is more compatible with the human immune system and exerts the least adverse effects. In this review, the comparative analysis of present polyvalent antivenom and future therapeutic protocols has been discussed.

Clinical and epidemiological profile of snakebite cases - A study from an industrial teaching hospital at Jamshedpur, Jharkhand, India.

India accounts for approximately half of the worldwide snakebite deaths. It is often a neglected public health problem and particularly in Jharkhand region where medical facilities are limited. Epidemiological and clinical profile-related studies are scarce. The present study aims to assess the epidemiological profile and clinical features of snakebites encountered in a tertiary-care teaching hospital at Jamshedpur, Jharkhand, India. The aim of this study was to assess the clinical profile, outcome and epidemiological factors of snakebite cases, admitted to a tertiary care hospital in Jamshedpur. This was a retrospective study from 2014 to 2021 wherein a total of 427 snakebite patients were admitted and had received treatment for snakebite at a tertiary-care teaching hospital at Jamshedpur, Jharkhand. All patients who reported with a history of snakebite were included in this study. The demographic and clinical details of each case were obtained and analysed. A total of 427 snakebite cases were admitted to the hospital during the study period. The victims were predominantly males. Majority of the bite cases encountered were from rural areas and were in the second quarter of the year. The site of the bite was largely on the lower limb and the upper limb had fewer bites. The Glasgow Coma Scale was normal in those who presented early. Acute kidney injury, neutrophilic leucocytosis and deranged liver enzymes were associated with bad prognosis. Timely intervention with anti-snake venom offered good result. We had more male patients (69.55%), belonging to rural areas (67.91%), more bites in lower limbs and more cases in the second quarter of the year. Mortality rate was 0.7%.