Duddingtonia flagrans冻干制剂厚壁孢子优化培养及其杀线虫幼虫剂量研究

发布时间：2018-03-17 21:22

  本文选题：Duddingtonia　切入点：flagrans　出处：《西北农林科技大学学报(自然科学版)》2017年09期 　论文类型：期刊论文

【摘要】：【目的】探究捕食线虫性真菌Duddingtonia flagrans冻干制剂厚壁孢子的批量培养方法及其对线虫幼虫的毒杀剂量。【方法】通过玉米粒或大麦粒培养基的单步培养法,以及先在含有0.05%琼脂粉的沙氏葡萄糖肉汤培养基中培养7d后继而转接到玉米粒或大麦粒培养基的双步培养法,以培养3~5周后洗脱的每克固体培养基中的厚壁孢子数为依据,对D.flagrans的最优培养方法进行筛选;然后使用最优培养法培养D.flagrans厚壁孢子并将其冻干后用于体外杀线虫幼虫试验,研究D.flagrans冻干制剂的杀线虫幼虫剂量。【结果】培养基质不同时,D.flagrans的菌落颜色有一定差异,玉米粒培养基中的菌落颜色呈微黄色,而大麦粒培养基中的菌落呈白色;D.flagrans的适宜培养时间为3周。采用单步培养法时,玉米粒培养基培养的厚壁孢子数量为2.4×10~5个/g,大麦粒培养基培养的厚壁孢子数量为3.2×10~5个/g;而采用双步培养法时,玉米粒培养基培养的厚壁孢子数量为3.0×10~5个/g,大麦粒培养基培养的厚壁孢子数量为3.5×10~5个/g。D.flagrans冻干制剂杀线虫幼虫剂量的阈值为每克粪便4×10~5个D.flagrans厚壁孢子,相应对线虫幼虫的杀虫率为95.2%。【结论】采用双步培养法培养厚壁孢子并将其制备成冻干制剂,在D.flagrans生物防治寄生性线虫病方面有较好的应用前景。
[Abstract]:[objective] to study the batch culture method of the freeze-dried preparation of nematodes, Duddingtonia flagrans, and its toxicity to nematode larvae. [methods] single step culture method was used to culture the nematode larvae in the medium of corn or wheat grains. And the two-step culture method was used to culture the medium containing 0.05% Agar powder for 7 days and then transferred to the medium of corn or wheat grains. The results were based on the number of thick parietal spores per gram of solid medium eluted after 35 weeks of culture. The optimum culture method of D. flagrans was screened, and the D. flagrans thick wall spores were cultured by the optimal culture method and then freeze-dried and used to kill nematode larvae in vitro. The dosage of D. flagrans freeze-dried preparation against nematode larvae was studied. [results] the colony color of D. flagrans was different in different culture medium, and the colony color of D. flagrans in cornkernel medium was yellowish. The optimum culture time of white D. flagrans in wheat medium was 3 weeks. The number of thick wall spores was 2.4 脳 10 ~ 5 / g in cornkernel medium and 3.2 脳 10 ~ 5 / g in wheat grain medium, while in the two-step culture method, the number of thick wall spores was 3.2 脳 10 ~ 5 / g. The number of thick wall spores was 3.0 脳 10 ~ 5 / g in cornkernel medium and 3.5 脳 10 ~ 5 / g 路D. flagrans in wheat medium. The threshold of larval dose of nematodes was 4 脳 10 ~ 5 D. flagrans per gram of feces, and the threshold value was 4 脳 10 ~ 5 D. flagrans thick wall spores per gram of feces. The killing rate of nematode larvae was 95.2%. [conclusion] using two-step culture method to culture thick wall spores and prepare them into freeze-dried preparation has a good application prospect in the field of D. flagrans biological control of parasitic nematode disease.
【作者单位】： 内蒙古农业大学兽医学院农业部动物疾病临床诊疗技术重点实验室;内蒙古自治区农牧业科学院兽医研究所;
【基金】：国家公益性行业(农业)科研专项(201303037) 内蒙古自然科学基金项目(2015MS0308) 家畜疫病病原生物学国家重点实验室开放基金项目(SKLVEB2015KFKT013)
【分类号】：S855.9
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