In Vitro Anti-Venom Potentials of Aqueous Extract and Oils of Toona ciliata M. Roem against Cobra Venom and Chemical Constituents of Oils
David Fred Okot*, Jane Namukobe, Patrick Vudriko, Godwin Anywar, Matthias Heydenreich, Oyedeji Adebola Omowumi, Robert Byamukama
There are high mortality and morbidity rates from poisonous snakebites globally. Many medicinal plants are locally used for snakebite treatment in Uganda. This study aimed to determine the in vitro anti-venom activities of aqueous extract and oils of Toona ciliata against Naja melanoleuca venom. A mixture of venom and extract was administered intramuscularly in rats. Anticoagulant, antiphospholipase A2 (PLA2) inhibition assay, and gel electrophoresis for anti-venom activities of oils were done. The chemical constituents of the oils of ciliata were identified using Gas chromatography-tandem mass spectroscopy ...(GC-MS/MS). The LD50 of the venom was 0.168 ± 0.21 µg/g. The venom and aqueous extract mixture (1.25 µg/g and 3.5 mg/g) did not cause any rat mortality, while the control with venom only (1.25 µg/g) caused death in 1 h. The aqueous extract of T. ciliata inhibited the anticoagulation activity of N. melanoleuca venom from 18.58 min. to 4.83 min and reduced the hemolytic halo diameter from 24 to 22 mm. SDS-PAGE gel electrophoresis showed that oils completely cleared venom proteins. GC-MS/MS analysis showed that the oils had sesquiterpene hydrocarbons (60%) in the volatile oil (VO) and oxygenated sesquiterpenes (48.89%) in the non-volatile oils (NVO). Some major compounds reported for the first time in T. ciliata NVOs were: Rutamarin (52.55%), β-Himachalol (9.53%), Girinimbine (6.68%) and Oprea1 (6.24%). Most compounds in the VO were reported for the first time in T. ciliata, including the major ones Santalene (8.55%) and Himachal-7-ol (6.69%). The result showed that aqueous extract and oils of T. ciliata have anti-venom/procoagulant activities and completely neutralized the venom. We recommend a study on isolation and testing the pure compounds against the same venom.
Results: Phytochemical screening of both aqueous and organic extracts of root barks of T. ciliata was performed, and the results showed that it contained various secondary metabolites, including alkaloids, tannins, saponins, phlorotannins, flavonoids, coumarins, anthraquinones, terpenoids, and glycosides. A detailed description of the phytochemicals has been published as a Mendeley data set vol. 1, 2021...
Conclusions: The compounds in aqueous extract and oils neutralized forest cobra (N. melanoleuca) venom. This was evidenced by the aqueous extract of T. ciliata neutralizing venom after Pre-incubation of venom with aqueous extract. Since aqueous extract causes blood cloth at some doses, it can be used as a procoagulant drug (a precursor of a natural substance necessary for the coagulation of blood), and it has the anticoagulation activity of venom close to antisera (standard drug). The volatile and non-volatile oils have anti-venom potential; therefore, they can be a good topical application as anti-venom for forest cobra bites. Generally, there has been no research performed on the root bark of Toona species. Therefore, aqueous extract and oils of T. ciliata showed high potential to be developed as anti-venom and aqueous extract as a coagulant herbal drug. Aroma is one of the most important attributes of food pharmaceuticals and is directly associated with product acceptance by consumers. T ciliata has a good yield of essential oil (3.10%) from the root bark. Therefore, this study provides research data for the application of aroma to pharmaceutical products of anti-venom drugs. We recommend future researchers make herbal formulations of medicine with this plant extract for anti-venom and procoagulant. The limitations of this study were the difficulty of hunting wild cobra, which was why we used cage ones, and SDS-PAGE was not showing all the proteins of forest cobra (N. melanoleuca) venom clearly.
Keywords: Snakebite; Venom; Medicinal plants; Anticoagulation; Antiphospholipase a2 inhibitor; Nutamarin and Seselin....
Medicinal plants species used by herbalists in the treatment of snakebite envenomation in Uganda
David Fred Okot*, Godwin Anywar, Jane Namukobe and Robert Byamukama
Background: There are high mortality and morbidity rates due to poisonous snakebites globally with sub-Saharan Africa having some of the highest cases. However, traditional medicine practitioners (TMP) have beentreating snakebites in Uganda for long despite the fact that few studies have been conducted to document such vital and rich indigenous traditional knowledge before it is lost. This study aimed to document the medicinal plant species used by experienced TMP in treating snakebite envenomation in selected post-conflict parts of Uganda. An ethnopharmacological survey was conducted in Kitgum, Serere, Kaberamaido and Kaabong districts in Uganda. ...Twenty-seven TMP with expertise in treating snakebites were purposively identified using the snowball technique and interviewed using semi-structured questionnaires. Data were analysed using simple descriptive statistics.
Results: Sixty plant species from 28 families were documented with high consensus among the isolated indigenous Ik tribe of Kaabong district. Most of the plant species used were from the Asteraceae and Fabaceae families with eight species each. The genus Echinops was the most well-represented with three species. The most commonly used plant species were of citation were Steganotaenia araliaceae (16), Microglossa pyrifolia (Lam.), Gladiolus dalenii Van Geel (13), Aframomum mildbraedii Loes. (11), Jasminum schimperi Vatke and Cyathula uncinulata (Schrad) Schinz (10) and Crinum macowanii Baker and Cyphostemma cyphopetalum (Fresen.) Desc. ex Wild & R.B. Drumm (10). S. araliaceae which was mentioned by all the TMP in the Ik community was used for first aid. Most of the plant species were harvested from the wild (68.75%) and were herbs (65.0%) followed by trees (23.3%). The most commonly used plant parts were roots (42.6%) and leaves (25.0%). Thirteen different methods of preparation and administration were used. Most of the medicines were administered orally (61.2%) and topically (37.6%). The commonest methods of oral application were cold water infusions (32.5%) and decoctions (21.7%).
Conclusions: TMP widely use several medicinal plant species for treating snakebite envenomation in the selected post-conflict regions of Uganda.
Keywords: Medicinal plants, Envenomation, Snakebite, Traditional medicine practitioners, Post-conflict, Uganda....