Ham P J, Townson S
Trans R Soc Trop Med Hyg. 1986;80(1):150-3. doi: 10.1016/0035-9203(86)90217-8.
A technique for improved cryopreservation at -196 degrees C of Brugia spp. microfilariae has been developed by modifications of a procedure previously used with Onchocerca spp. A double incubation in ethanediol (ED) solutions, firstly at 37 degrees C in 10% (v/v) ED for 15 min and secondly at 0 degrees C in 40% (v/v) ED for 45 sec followed by plunging into liquid nitrogen, resulted in over 90% of the microfilariae of B. malayi exhibiting normal motility. When used with B. pahangi microfilariae, followed by blood-feeding to Aedes aegypti (Bels New strain) a third-stage larvae yield of 4.04 per fly was obtained, compared with 5.16 in the unfrozen group. This represents an infectivity level of 78% compared with unfrozen controls. The technique also has the benefits of being suitable for field conditions, requiring no sophisticated equipment, merely some ice, ethanediol and liquid nitrogen.
通过对先前用于盘尾丝虫属的程序进行修改，已开发出一种在-196℃下改进马来丝虫微丝蚴冷冻保存的技术。在乙二醇（ED）溶液中进行两次孵育，首先在37℃下于10%（v/v）ED中孵育15分钟，其次在0℃下于40%（v/v）ED中孵育45秒，然后投入液氮，结果超过90%的马来丝虫微丝蚴表现出正常活力。当用于彭亨丝虫微丝蚴时，随后将其喂给埃及伊蚊（贝尔斯新菌株），每只蚊子获得的第三期幼虫产量为4.04条，而未冷冻组为5.16条。与未冷冻的对照组相比，这代表了78%的感染率。该技术还具有适合野外条件的优点，不需要复杂的设备，只需要一些冰、乙二醇和液氮。
