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消费者表面消毒对自然环境中陆生无脊椎动物饮食DNA宏条形码数据及饲养试验的影响

Effects of consumer surface sterilization on diet DNA metabarcoding data of terrestrial invertebrates in natural environments and feeding trials.

作者信息

Miller-Ter Kuile Ana, Apigo Austen, Young Hillary S

机构信息

Department of Ecology, Evolution, and Marine Biology University of California Santa Barbara Santa Barbara CA USA.

出版信息

Ecol Evol. 2021 Aug 9;11(17):12025-12034. doi: 10.1002/ece3.7968. eCollection 2021 Sep.

Abstract

DNA metabarcoding is an emerging tool used to quantify diet in environments and consumer groups where traditional approaches are unviable, including small-bodied invertebrate taxa. However, metabarcoding of small taxa often requires DNA extraction from full body parts (without dissection), and it is unclear whether surface contamination from body parts alters presumed diet presence or diversity.We examined four different measures of diet (presence, rarefied read abundance, richness, and species composition) for a terrestrial invertebrate consumer (the spider ) both collected in its natural environment and fed an offered diet item in contained feeding trials using DNA metabarcoding of full body parts (opisthosomas). We compared diet from consumer individuals surface sterilized to remove contaminants in 10% commercial bleach solution followed by deionized water with a set of unsterilized individuals.We found that surface sterilization did not significantly alter any measure of diet for consumers in either a natural environment or feeding trials. The best-fitting model predicting diet detection in feeding trial consumers included surface sterilization, but this term was not statistically significant ( = -2.3, -value = .07).Our results suggest that surface contamination does not seem to be a significant concern in this DNA diet metabarcoding study for consumers in either a natural terrestrial environment or feeding trials. As the field of diet DNA metabarcoding continues to progress into new environmental contexts with various molecular approaches, we suggest ongoing context-specific consideration of the possibility of surface contamination.

摘要

DNA宏条形码技术是一种新兴工具,用于量化传统方法不可行的环境和消费群体中的食物组成,包括小型无脊椎动物类群。然而,对小型类群进行宏条形码分析通常需要从整个身体部位(不解剖)提取DNA,目前尚不清楚身体部位的表面污染是否会改变假定的食物组成或多样性。我们使用对整个身体部位(腹部)进行DNA宏条形码分析的方法,研究了一种陆生无脊椎动物消费者(蜘蛛)在自然环境中采集的以及在封闭式喂食试验中喂食特定食物后的四种不同的食物组成测量指标(存在情况、稀疏读取丰度、丰富度和物种组成)。我们将在10%商业漂白剂溶液中表面灭菌以去除污染物,然后用去离子水冲洗后的消费者个体的食物组成,与一组未灭菌个体的食物组成进行了比较。我们发现,无论是在自然环境还是喂食试验中,表面灭菌都没有显著改变消费者的任何食物组成测量指标。预测喂食试验消费者食物检测情况的最佳拟合模型包括表面灭菌,但该因素在统计学上并不显著(β = -2.3,P值 = 0.07)。我们的结果表明,在这项DNA食物宏条形码研究中,无论是在自然陆地环境还是喂食试验中,表面污染似乎都不是一个重大问题。随着食物DNA宏条形码领域继续通过各种分子方法进入新的环境背景,我们建议针对具体情况持续考虑表面污染的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54fd/8427582/e8668ff88827/ECE3-11-12025-g005.jpg

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