粪便显微分析和DNA宏条形码技术在野猪食性分析中的应用和比较

Application and Comparison of Fecal Microscopic Analysis and DNA Metabarcoding Technology for Dietary Analysis of Wild Boar

有蹄类动物食性分析对基础生态学研究乃至大型濒危食肉动物保护与恢复有重要科学价值。食物组成定量化有很多方法,常用的方法有粪便显微分析和DNA宏条形码技术,但各有其优缺点。2014年12月—2016年3月,在珲春、汪清、黄泥河、天桥岭和穆棱5个地区收集了野猪(Sus scrofa)新鲜粪便样品,运用粪便显微分析和DNA宏条形码技术比较研究了冬季野猪食性。仅对比分析了野猪植物性食物,通过粪便显微分析获得的野猪食物组成包含32属植物,比例大于4%的主要食物有木贼属(Equisetum,(22.76±1.87)%)、松属(Pinus,(22.56±1.73)%)、薹草属(Carex,(25.41±2.09)%)和苔藓(Bryophyta,(4.95±1.02)%);通过DNA宏条形码技术分析获得的野猪食物组成包含60属植物,比例大于4%的主要食物有薹草属(30.02±5.95)%、唐菖蒲属(Gladiolus,(18.76±4.12)%)、胡桃属(Juglans,(6.63±3.84)%)、蚊子草属(Filipendula,(5.23±2.79)%)、榛属(Corylus,(4.23±1.60)%)和绣线菊属(Spiraea,(4.93±4.06)%)。通过分析野猪食物多样性,珲春、汪清和穆棱地区粪便显微分析显著高于DNA宏条形码技术分析得到的食物物种丰富度,黄泥河地区粪便显微分析显著低于DNA宏条形码技术分析得到的Shannon-Wiener指数和生态位宽度,珲春地区粪便显微分析显著高于DNA宏条形码技术分析得到的食物均匀度指数。粪便显微分析和DNA宏条形码技术分析得到的食物选择种类和比例具有一定的差异性和一致性,其中,DNA宏条形码技术可分析得到更多的草本类食物种类。因此,建议采用粪便显微分析法时应综合考量采食范围与采食情况,全面分析出采食种类和比例;采用DNA宏条形码技术时应尽量完善目标动物食源DNA条形码数据库,准确分析出目标动物在活动范围内采食种类。

Diet analysis of ungulate is of great scientific value for the study of basic ecology and the protection and restoration of large endangered carnivores.There are many methods to quantify food composition,including fecal microscopic analysis and DNA metabarcoding technology,but each has its own advantages and disadvantages.From December 2014 to March 2016,fresh fecal samples were collected from Hunchun,Wangqing,Huangnihe,Tianqiaoling and Muling,and the diet of wild boar(Sus scrofa)in winter was comparatively studied by fecal microscopic analysis and DNA metabarcoding technology.In this study,only the plant food of wild boars was compared,and the results showed using fecal microscopic analysis,the dietary composition of wild boar included 32 genera of plants.In general,the main food of wild boar with the proportion of various plants more than 4%is Equisetum(22.76±1.87)%,Pinus(22.56±1.73)%,Carex(25.41±2.09)%,Bryophyta(4.95±1.02)%.On the other hand,using DNA metabarcoding technology,the dietary composition of wild boar included 60 genera of plants.In general,the main food of wild boar with the proportion of various plants more than 4%is Carex(30.02±5.95)%,Gladiolus(18.76±4.12)%,Juglans(6.63±3.84)%,Filipendula(5.23±2.79)%,Corylus(4.23±1.60)%,and Spiraea(4.93±4.06)%.By analyzing the dietary diversity of the wild boar,species richness index was significantly higher in Hunchun,Wangqing and Muling areas using fecal microscopic analysis approach than using DNA metabarcoding technology.The Shannon-Wiener index and niche breadth obtained by DNA metabarcoding technology were significantly lower than those obtained by fecal microscopic analysis in Huangnihe area.The species evenness index of wild boar in Hunchun area was significantly higher by fecal microscopic analysis than by DNA metabarcoding technology.Therefore,to some extent the species and proportion obtained by fecal microscopic analysis and DNA metabarcoding technology showed consistent outcomes but also generated differences.DNA metabarcoding technology could identify even greater number of herbaceous species.Therefore,we suggested that the usage of fecal microscopic analysis approach should comprehensively consider ungulate feeding range and situation,so that the species and feeding proportion could be more accurately analyzed.When using DNA metabarcoding technology,we should improve DNA barcode database of the food sources of targeted animals,which may analyze food species more accurately within their distribution ranges.