الملخص الإنجليزي
Abstract
Background: Envenomation by snakebites is a serious crisis that covers almost the whole globe, especially, Asia and Africa. This leads to either morbidity (with deformation of skin and muscles leading to disability after recovery from necrosis and haemorrhage) and/or mortality. Local haemorrhage is an important manifestation due to snake envenoming with no known treatment, Unfortunately, the only approved therapy available is the use of commercialized anti-venom even though it is not efficient to neutralize local haemorrhage. Objectives: To evaluate the efficacy of Phoenix dactylifera L. Ajwa date fruit extract in neutralizing local haemorrhage induced by three clinically important snakes' venoms. Furthermore, to isolate the active components from the Ajwa date fruit and examine each of which for its local haemorrhage neutralization capacity. Methodology& Results: Ajwa date fruit was extracted using three solvents and extracts were examined for their anti-local haemorrhage activity using in vitro and in vivo assays. Only the crude ethanolic extract (CEE), was found to be potentially effective both in vitro and in vivo in neutralizing the local hemorrhagic effects induced by venoms obtained from three clinically important snakes (Bothrops jararaca, Crotalus atrox and Echis carinatus). The CEE was fractionated using High Performance Liquid Chromatography (HPLC) and 140 different fractions (Frc) were obtained. These fractions were examined for their anti local haemorrhage and anti-lethal activities. Only four out of the 140 fractions were found to be significantly effective [namely; Frc24, Frc25, Frc28 & Frc32] against individual and pooled venoms of the three snakes' venoms together using both in vitro and in vivo assays and analyzed using Mann-Whitney analysis with p<0.05. Characterization and identification of the isolated active fractions was performed using the Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF) and mass bank searches. Only Frc25 showed peaks, whereas the other fractions were found difficult to be processed further due to lacking of purity. Mass bank search analysis of Fro25 showed two peaks and identified as: (i) Steroids and (ii) Flavonoid, Phytochemicals test showed that both the CEE and the four fractions (Frc24, Frc25, Fro28 and Frc32) contain carbohydrates (glucose) only and no other constituents. The active fractions as well as the CEE were further examined for their anti-lethality (LD100) in vivo against pooled venoms [Bothrops jararaca (30mg/0.03kg), Crotalus atrox (50mg/0.03kg) and Echis carinatus (50mg/0.03kg)] and the results found to be potentially significant by elongating the time of survival of the tested animals in comparison to that of the control animals (P<0.05). Furthermore, the optimum concentrations of the CEE for the anti-lethality of LDso test using 27.5mg/0.03kg of B. jararaca venom were significant (P<0.05) at concentrations of 200 and 300 u1/0.03kg. Conclusion: The active components from the Phoenix dactylifera L Ajwa date fruit that have the capacity to neutralize the local haemorrhage were isolated. These isolated active components shows both in vitro and in vivo the capacity to cross react and neutralize 100% of the local haemorrhage effects caused by the three clinically important snake venoms
