家蚕BmNPV压力选择品系的细菌病发生及其病原分离鉴定  

Bacterial disease occurrence and pathogen isolation and identification of BmNPV pressure selected strains in silkworm(Bombyx mori)

作　　者：邵榆岚[1] 范永慧[1] 范仕弘 张永红[1] 张一川[1] 白兴荣[1] SHAO Yu-lan;FAN Yong-hui;FAN Shi-hong;ZHANG Yong-hong;ZHANG Yi-chuan;BAI Xing-rong(Sericulture and Apiculture Research Institute,Yunnan Academy of Agricultural Sciences,Honghezhou,Yunnan 661101,China)

机构地区：[1]云南省农业科学院蚕桑蜜蜂研究所,云南红河州661101

出　　处：《南方农业学报》2023年第11期3388-3395,共8页Journal of Southern Agriculture

基　　金：现代农业产业技术体系建设专项(CARS-18-SYZ21);云南省重大科技专项(202102AE090010)。

摘　　要：【目的】调查鉴定核型多角体病毒(BmNPV)压力选择家蚕相关品系细菌病发病原因,为蚕作生产上家蚕抗Bm NPV品种细菌病防治提供理论依据。【方法】相同批次和饲养条件下,对BmNPV压力选择自建品系P50Nn(n表示代)和抗Bm NPV品种4个母种品系苏N、菊N、明N、虎N的正常保种组发生家蚕细菌病情况进行初步调查,分离培养病蚕肠道细菌,开展16S rDNA分子鉴定、革兰氏染色和扫描电镜等形态学鉴定,以及糖发酵、吲哚试验等20项生化鉴定。【结果】细菌病导致家蚕BmNPV压力选择品系添食BmNPV第3代次的P50N3结茧率为47.63%,抗BmNPV育种母种品系添食BmNPV多代次的菊N结茧率最低为34.72%,而未添食过BmNPV的对照品系P50结茧率为96.15%,P50N3和菊N结茧率与P50呈极显著差异(P<0.01)。病蚕肠道细菌经过多次分离纯化,显微镜观察球状细菌较多;分子鉴定后获得肠球菌属(Enterococcus)、葡萄球菌属(Staphylococcus)、柠檬酸杆菌属(Citrobacter)、沙雷氏菌属(Serratia)、克雷伯氏菌属(Klebsiella)、假单胞菌属(Pseudomonas)和肠杆菌属(Enterobacter)等7类细菌属,其中葡萄球菌属占比(30.00%)最高,其次为肠球菌属和沙雷氏菌属,均为16.67%。选择分子鉴定一致性最高的8株不同细菌,进一步进行形态学鉴定和生化鉴定,与分子鉴定结果一致。【结论】家蚕BmNPV压力选择品系的细菌病发生主要由葡萄球菌、肠球菌和多种肠杆菌科细菌引起,在BmNPV压力选择早期家蚕细菌病发生更严重。【Objective】To investigate and identify the causes of bacterial diseases in the Bombyx mori nucleopolyhe‐droviurs(BmNPV)pressure selection related strains of silkworms,in order to provided a theoretical basis for the preven‐tion and control of BmNPV resistant bacterial diseases in silkworm production.【Method】Under the same batch and feeding conditions,a preliminary investigation was conducted on the occurrence of bacterial diseases in the normal group of BmNPV pressure selective strain P50Nn(n indicated generation)and four female lines Su N,Ju N,Ming N and Hu N of anti-BmNPV breeds.The intestinal bacteria of the diseased silkworms were isolated and cultured.Then carried out 16S rDNA molecular identification,morphological identification such as Gram staining and scanning electron microscopy,and 20 biochemical identifications including sugar fermentation and indole test.【Result】Bacterial diseases caused the co‐cooning rate of P50N3 in the BmNPV pressure selection strain of silkworms fed with BmNPV for the third generation to be 47.63%,the cocooning rate of JU N in the mother strain of BmNPV resistant variety that fed with BmNPV for multiple generations was the lowest at 34.72%,while the cocooning rate of P50 in the control strain without BmNPV was 96.15%,the cocoon formation rates of P50N3 and JU N were extremely significantly different from those of P50(P<0.01).The in‐testinal bacteria of diseased silkworms had been separated and purified for multiple times,and there were many spherical bacteria observed under the microscope.After molecular identification,seven types of bacteria were obtained,including Enterococcus,Staphylococcus,Citrobacter,Serratia,Klebsiella,Pseudomonas and Enterobacter,among which Staphylococcus accounted for the highest proportion of 30.00%,followed by Enterococcus and Serratia,both accounting for 16.67%.The 8 different bacterial strains with the highest consistency in molecular identification were selected for fur‐ther morphological and biochemical identification,which w

关 键 词：病毒压力 家蚕抗性 细菌病 病原鉴定 

分 类 号：S884[农业科学—特种经济动物饲养]