摘要
感染家蚕微孢子虫的成品卵检测是业内关注的技术问题之一。用磨碎管分装1000粒蚕卵并加海绵塞,进行一段式催青的实验室二日孵化率达98.25%或以上,对生产用不同蚕品种、越年或冷藏浸酸蚕种及不同单位生产蚕种的二日孵化率最低为94.40%。用含0.01%十二烷基磺酸钠和2.00%氢氧化钠的溶液浸泡和磨碎蚕卵样本(约1000粒),固形物去除率为83.38%。5龄起蚕添食感染5×10^(5)颗/mL家蚕微孢子虫孢子悬浮液后获取的混合蚕卵,在催青孵化后第7天检出率明显上升。1000粒蚕卵混入3000颗家蚕微孢子虫孢子的检出率为100%。蚕卵磨碎液用PBS缓冲液(pH=7.2~7.6)稀释40倍或以上时,qPCR法检测可稳定检出家蚕微孢子虫,检测灵敏度为4×10^(6)颗/mL,低于光学显微镜法的1.5×10^(3)颗/mL。105个一代杂交蚕种母蛾检疫批(166623张毛种)母蛾检测和蚕卵集团检测的检出情况类似。不论是母蛾检测还是成品卵检测都涉及家蚕微孢子虫的胚传率和流行病学风险管控的阈值确定,上述研究提供了一种成品卵集团检测的方法,但实际应用尚需更为丰富的流行病学调查数据。
The pebrine spore(Nosema bombycis)detection of commercial silkworm eggs is one of the technical issues of concern in the sericultural industry.In this study,1000 silkworm eggs were packed in grinding tubes with sponge plugs,to conduct a one⁃stage incubation experiment in the laboratory,with a two⁃day hatchability of 98.25%or higher.The minimum two⁃day hatchability for different silkworm varieties,hibernating eggs,acid⁃treated eggs after refrigeration,and silkworm eggs from different production units,was 94.40%.For the egg samples(about 1000 eggs)soaked and ground with 0.01%dodecyl sodium sulfonate and 2%sodium hydroxide solution,the solid removal rate was 83.38%.The detection rate of the mixed eggs obtained from the 5th instar silkworm after feeding with 5×10^(5)spores/mL of N.bombycis increased significantly on the 7th day after incubation.For 1000 eggs mixed with 3000 N.bombycis spores,the detection rate was 100%.When the grinding solution of silkworm eggs was diluted 40×or more with PBS buffer solution(pH=7.2-7.6),the N.bombycis could be stably detected with the qPCR method,and the detection sensitivity was 4×10^(6)spores/mL,which was lower than that of the optical microscopy method(1.5×10^(3)spores/mL).The detection results of mother moths and eggs group inspection were similar in 105 inspection lots of mother moths(including 166623 sheets of unselected loose egg)from the first⁃generation hybrid silkworm.The detection of both mother moths and commercial eggs involves the determination of embryo transmission rates and epidemiological risk control thresholds for silkworms.This study provides a method for the group inspection of commercial eggs,while more epidemiological survey data are needed for further application.
作者
朱春群
张宪翠
何恩洁
范明亮
孟正乐
楼霞
邵勇奇
鲁兴萌
Zhu Chunqun;Zhang Xiancui;He Enjie;Fan Mingliang;Meng Zhengle;Lou Xia;Shao Yongqi;Lu Xingmeng(Zhejiang Agricultural Technology Extension and Service Center,Hangzhou 310019,China;College of Life Science,Huzhou University,Huzhou Zhejiang 313000,China;Hangzhou Sericulture Technology Promotion Station,Hangzhou 310017,China;The Rural Development Academy,Zhejiang University,Hangzhou 310058,China)
出处
《蚕业科学》
CAS
CSCD
北大核心
2023年第5期414-422,共9页
ACTA SERICOLOGICA SINICA
基金
浙江省“三农九方”科技协作计划项目(2022SNJF041)。
关键词
家蚕
家蚕微孢子虫
成品卵
集团
检测
Bombyx mori
Pebrine spore
Commercial egg
Group
Inspection
