基于k-means聚类算法的兴义维蚋幼虫龄数的估算
Estimation of larval instar of Simulium xingyiense based on k-means clustering algorithm
赵娜 1王毅 1杨曜铭 1吴慧 1修江帆 1寻慧 1杨明1
作者信息
- 1. 贵州医科大学基础医学院生物学系&医学昆虫重点实验室,贵州贵阳 550025
- 折叠
摘要
目的 探讨基于k-means聚类算法估算兴义维蚋幼虫的龄数,以明确虫龄与日龄及鳃斑发育阶段的关系.方法 采集贵州青岩河流中兴义维蚋虫卵,于实验室的蚋类饲养系统中培育至幼虫,每日收集幼虫至大量化蛹,持续20 d;收集到幼虫1 112头,于体视显微镜下测量头壳长(HCW)、后颊长(PGL)、上颚基横骨长(MPL)及体长(BL),观察不同虫龄幼虫的破卵器、鳃斑及性腺的形态学特征,并判断其龄期、分析虫龄与日龄和鳃斑发育关系;选取具有明显腮斑发育的兴义维蚋幼虫10头、制作石蜡切片,采用HE染色鉴定精巢和卵巢、辅助确认性别鉴定结果;采用k-means聚类方法划分形态计量学数据、利用Brooks-Dyar定律检测k-means聚类结果,R语言下行聚类与Brooks-Dyar定律检验、t检验等,根据拟合度分析判断兴义维蚋幼虫龄数.结果 1 112头兴义维蚋幼虫中有破卵器89头,出现鳃斑发育334头,6～7龄幼虫320头,有预蛹特征34头;Brooks-Dyar定律与破卵器、鳃斑形态特征显示7龄幼虫假设符合昆虫幼虫生长规律;组织学观察见幼虫精巢为椭圆形、体积大、外有几丁质层包裹,卵巢为长条形、体积小、后端有色素细胞包裹;鉴定6、7龄幼虫性别结果,仅7龄幼虫雌雄性PGL有差异(P＜0.05);幼虫虫龄与日龄、鳃斑发育关系结果显示,约2～3 d对应1个幼虫生长龄期,第17天首见幼虫蛹化,6龄幼虫出现明显的鳃斑.结论 兴义维蚋幼虫具7龄,实验室下幼虫发育约需3周,最短17 d;腮斑发育起始于6龄,7龄出现明显形态学特征.
Abstract
Objective To explore the estimation of larval instars of Simulium xingyiense based on k-means clustering algorithm to clarify the association of larval instars with day age and the stage of gill spot development.Methods The eggs of Simulium xingyiense in Qingyan River,Guizhou province were collected and cultured in the Simuliidae feeding system until they became larvae.A large number of larvae were collected daily until they pupated for 20 days.A total of 1,112 larvae were collected.A stereomicroscope was used to measure head capsule width(HCW),postgenal length(PGL),length of the mandibular phragma(MPL),and body length(BL).The morphological characteristics of the egg bursters,gill spots and gonads of the larvae at different instars were recorded to judge larvae instars.The relationship between instars and day age and the development of gill spots was analyzed.Ten larvae of Simulium xingyiense with obvious gill spot development were selected and prepared for paraffin sections.HE staining was performed to identify testes and ovaries,and assist in confirming gender identification results.k-means clustering was used to partition morphometric data.Brooks Dyar's law was used to examine k-means clustering results.Under R language,clusteringand Brooks Dyars law test,t-test,etc.were performed.Larval age of Simulium xingyiense was judged based on fitting degree analysis.Results Out of 1,121 larvae of Simulium xingyiense,there were 89 larvae with egg bursters,334 larvae with gill spot development,320 larvae with 6th to 7th instar larvae and 34 larvae with pre-pupal characteristics.Brooks Dyar's law,the morphological characteristics of the egg bursters and gill spots exhibited that the hypothesis of 7th instar larvae conforms to the growth pattern of insect larvae.Histological observation shows that the larval testes were ellipse,large volume and surrounded by a chitin layer on the outside,while the ovaries were elongated in shape,small volume and surrounded by pigment cells at the posterior end.The results of identifying the gender of 6th and 7th instar larvae showed that there was only a difference in PGL between male and female 7th instar larvae(P＜0.05).The association of larval age with day age and gill spot development showed that approximately 2 to 3 days corresponded to one larval growth stage.On the 17th day,the pupation was first observed in the larvae.The obvious gill spots appeared in the 6th instar larvae.Conclusion The larvae of Simulium xingyiense have 7 instars,and the development of larvae in the laboratory takes about 3 weeks with a minimum of 17 days.The development of gill spots begins at the 6th instar,and obvious morphological features appear at the 7th instar.
关键词
蚋科/组织学/性腺/龄数/形态计量学/k-means聚类Key words
Simuliidae/histology/gonad/instar/morphometry/k-means clustering引用本文复制引用
基金项目
国家自然科学基金(31860324)
贵州医科大学国家自然科学基金培养项目(20NSP018)
出版年
2024
NETL
NSTL
万方数据