Objective: To establish an animal model of sparganosis mansoni through oral administration of Cyclops infected with procercoids.
Methods: Domestic cats were infected with Sparganum mansoni under laboratory conditions, and fresh cat stool samples were collected, washed in dechlorinated water, and filtered. Spirometra mansoni eggs were collected and prepared into suspensions. Twenty C57BL/6j mice were randomly divided into the experimental group (n = 15) and the control group (n = 5). Wild Cyclops were infected with Spirometra mansoni coracidia to allow 3 to 5 procercoids in each Cyclop. Then, each mouse in the experimental group was given 15 Cyclops infected with procercoids by gavage, while mice in the control group were orally administered with the same volume of dechlorinated water. All mice were sacrificed after 5 months, and dissected, and suspicious Sparganum mansoni worms were collected. The serum specific IgG antibody against Sparganum mansoni was measured in mice using enzyme-linked immunosorbent assay (ELISA). Genomic DNA was isolated from suspicious Sparganum mansoni worms, and the specific Sparganum mansoni cytochrome oxidase I (COI) gene was amplified using PCR assay.
Results: Among the 15 mice in the experimental group, six were positive for the serum specific IgG antibody against Sparganum mansoni, and milky white worms were found and collected from the subcutaneous regions of 4 out of 6 mice. Only one worm was detected in each mouse, and the worm morphology was similar to Sparganum mansoni. Capillary electrophoresis of the PCR amplification products of COI gene presented a specific band with 151 bp in size, and sequencing analysis revealed 100% homology with Sparganum mansoni.
Conclusions: A mouse model of sparganosis mansoni is successfully created through oral administration of Cyclops infected with Spirometra mansoni procercoids.
［摘要］ 目的 探索通过感染原尾蚴的剑水蚤灌胃小鼠建立曼氏裂头蚴病小鼠动物模型的可行性。方法 实验室条件 下用曼氏裂头蚴感染家猫, 收集当日新产猫粪, 用去氯水淘洗后过滤获得曼氏迭宫绦虫虫卵, 制成虫卵悬浊液。将 20 只 C57BL/6j 小鼠随机分为实验组和对照组, 实验组 15 只、对照组 5 只。以曼氏迭宫绦虫钩球蚴感染野生剑水蚤, 使每只剑 水蚤体内含有 3 ~ 5 条原尾蚴; 再将感染原尾蚴的剑水蚤灌胃实验组小鼠, 每只小鼠灌入剑水蚤 15 只, 对照组灌胃等体积 去氯水。6 个月后处死小鼠, 分离疑似裂头蚴虫体, 并用间接酶联免疫吸附试验检测小鼠血清中抗曼氏裂头蚴特异性 IgG 抗体水平。提取疑似虫体基因组 DNA, 以曼氏裂头蚴细胞色素 C 氧化酶 1 (COI) 基因片段为目的序列进行 PCR 扩增 以鉴别疑似虫体。结果 实验组 15 只小鼠中, 6 只小鼠血清抗曼氏裂头蚴特异性 IgG 抗体检测呈阳性; 于其中 4 只小鼠 皮下发现并分离出乳白色虫体。每只小鼠均仅检出 1 条虫体, 虫体形态与曼氏裂头蚴相似。基于 COI 基因的疑似虫体 PCR 扩增产物在毛细管电泳上于 151 bp 处显示特异性条带, 测序结果与已知曼氏裂头蚴序列 100% 符合。结论 本研究 成功建立了一种曼氏裂头蚴病小鼠动物模型。.
Keywords:
Animal model; Procercoid; Sparganosis mansoni;