Collection, Bioactivities And Proteomic Analysis Of The Crude Venom From The Spider L.Tredecimguttatus

The spider Latrodectus tredecimguttatus , also called black widow spider, belongs to the family of Theriodiidae. It is a medium-sized spider and is one of the most poisonous spiders in the world.In the present study, the spiders L tredecimgattatus were successfully indoor-reared. The morphology, living inhabits and ecological behaviors of the spider were observed and investigated. At the same time, the effects of temperature, humidity and food on the activities, survival rate and fecundity were also investigated.The statistical analysis indicates that the average lengths of female and male spider are 10.37mm and 4. 58mm, respectively; and the average body weights are 0.475g and 0. 104g , respectively. The average number of egg bag laid by a female spider is 4. About 200-300 baby spiders hatched from an egg bag. Baby female and male spiders grew into adult spiders after 6-7 peelings and 4 peelings, respectively. Most adult spiders died before the beginning of the winter. Life spans of female and male spider are 130~300d and 98~144d, respectively. When female spider mated with the male spider that hatched about 30d later, the laying egg rate was higher than that when female spider mating the male spider that hatched in the same period as the female spider. Staying in egg bag for a certain period is necessary for baby spiders'survival and development. Relative humidity is the most important factor for the indoor-rearing of the spider. The relative humidity of about 65% and indoor temperature of about 25°C are the optimized conditions for the indoor-rearing.Several methods for venom collection, including electrical stimulation and dissecting venom gland, were compared and the results showed that electrical stimulation method was the best. The composition of venom obtained by electrical stimulation and dissecting venom gland were comparatively studied using RP-HPLC, mass spectrometry and two-dimensional gel eletrophoresis.The intraperitoneal LD5o in mice of the venom collected by electrical stimulation and dissecting venom gland were 0. 164mg/kg and 0.393mg/kg, respectively. Electrical physiological experiments indicated that both of the venoms could block neuromuscular transmission in isolated mouse phrenic nerve-diaphragm preparation. Venom collected by electrical stimulation could block the neuromuscular transmission completely in 23. 3 + 2. 2min at the concentration of 3. 2 ii g/ml. A 6u g/ml of venom obtained by dissecting venom gland could block the neuromuscular transmission completely in 25. 0±2. 2min, which indicated that the biological activity of venom collected by electrical stimulation was higher that of venom collected by dissecting venom gland.The venom collected by dissecting venom gland was separated by two-dimensional gel electrophoresis (2D-PAGE). Image analysis showed that the venom was rich in acidic...