Effect of Escherichia coli endotoxin on Archachatina marginata hemolymph coagulation system.

ABSTRACT

CONTEXT Archachatina marginata Swainson (Achatinidae) is found in Nigeria, West Africa. Its hemolymph is applied as a disinfectant to blades and fresh cuts of circumcision in Yorubaland. The hemolymph is also used in traditional medicine practice. Investigation into its anti-endotoxin response is being studied for the first time. OBJECTIVE: This study determined whether endotoxin causes measurable and concentration-dependent protein coagulation in the separate hemolymph fractions and in hemocyte lysate (HL)/plasma mixtures. MATERIALS AND METHODS: Endotoxin was prepared by inoculating 5% w/v dextrose with locally isolated Escherichia coli cells and incubated for 48 h before sterilization. Pyrogenicity was determined by rabbit test method and use the of LAL kit. Hemolymph fractions were exposed to endotoxin while controls were exposed to endotoxin-free water (0.025 EU/ml). HL/plasma (11 v/v) was exposed to varied endotoxin concentrations. RESULTS: Data indicated significantly higher protein coagulates induced by endotoxin in all the hemolymph fractions (P < 0.05). Maximum protein coagulation in mixture of HL/plasma 11 was recorded. Exposure of HL/plasma at optimal ratio to varied endotoxin caused linear protein coagulation up to 1.0 EU/ml, beyond which it dropped significantly and unresponsive to further increase in endotoxin doses. DISCUSSION AND CONCLUSION: There was endotoxin-induced protein coagulation, which is endotoxin concentration-dependent. The optimal coagulation observed for 11 HL/plasma mixture suggests stronger interaction between the hemocytes and the plasma in response to endotoxin. There are LPS-binding proteins in the plasma and hemocytes of A. marginata. This finding may be employed in detection and quantification of endotoxin in future.