Limulus amoebocyte lysate (LAL) clots when mixed with of endotoxins which are lipopolysaccharide components of the outer cell-wall layer of gram-negative bacteria. Since the initial description of the lysate method for the detection of endotoxins, it has become an international standard method. Although Jinmen and the Penghu coast has the wild Chinese horseshoe crab (Tachypleus tridentatus), but not yet developed into the commercialization of biological agents. This study investigated the horseshoe crab blood into the appropriate reagent season, adequate blood volume, feeding withy β-1,3-glucan on horseshoe crab and blood composition, deformation of cell lysate TAL endotoxin sensitivity before and after extracting the differences and TAL antibacterial test blood components.
Results: When the water temperature more than 20 ℃,The amoebocytes of horseshoe crab increased gradually since April, started to increase sensitivity to endotoxin, as appropriate sampling blood time. Horseshoe crab of blood volume weight about 26%, the appropriate sampling blood volume to be less than 5% body weight horseshoe crab can improve bio-security and reduced mortality. β-1,3-glucan in the feeding period, although the Amoebocyte can increase Limulus enhance sensitivity to endotoxin, but after stopping feedingβ-1,3-glucan, the sensitivity to endotoxin did not vote than β-1,3-glucan with low sensitivity. Amoebocyte were extracted, the sensitivity can reach 0.015EU, relative to the extraction without the TAL, 0.015EU endotoxin concentration can improve the sensitivity of 100%. Experimental freeze-drying preservation at present can be maintained for 3 months endotoxin agglutination test of 0.6EU. Horseshoe crab Limulus detected in serum and cell lysate made from 17 kinds of antimicrobial piece, of two Vibrio alginolyticus, E. coli and one Streptococcus, no antibacterial effects.