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揭示鲑鱼虱的浮游阶段:一种独特的荧光信号,可快速识别浮游动物组合中的罕见桡足类。

Illuminating the planktonic stages of salmon lice: A unique fluorescence signal for rapid identification of a rare copepod in zooplankton assemblages.

机构信息

Institute of Marine Research, Bergen, Norway.

University of Stirling, Stirling, Scotland.

出版信息

J Fish Dis. 2021 Jul;44(7):863-879. doi: 10.1111/jfd.13345. Epub 2021 Feb 15.

Abstract

Monitoring of planktonic salmon louse (Lepeophtheirus salmonis salmonis) abundance and parameterization of key life-history traits has been hindered by labour-intensive and error-prone quantification using traditional light microscopy. Fluorescence illumination has been proposed as a means of improving visualization, but prior to this study adequate investigation of the relevant fluorescence profiles and measurement conditions has not been undertaken. We investigated the fluorescence profiles of L. salmonis and non-target copepod spp. with excitation and emission matrices (200-600 nm) and identified unique fluorescence signals. Fluorescence microscopy using excitation wavelengths of 470 ± 40 nm, and emission wavelengths of 525 ± 50 nm, showed that after 90 days of formalin storage salmon lice have a mean fluorescence intensity that is 2.4 times greater than non-target copepods (copepodid and adult stages). A 7-day heat treatment of 42°C in formalin increased the difference between salmon louse copepodids and non-target copepods to a factor of 3.6, eliminating the need for prolonged storage. Differences in the fluorescence signal and endogenous fluorophores were investigated with respect to variation in sea lice species, age, stage and host fish origin. Under the conditions outlined in this paper, the fluorescence signal was found to be a reliable means of visualizing and differentiating salmon lice from non-target zooplankters. Adaptation of the fluorescence signal would greatly expedite traditional methods of enumerating salmon louse larvae in plankton samples and could provide a means of automated detection.

摘要

浮游性鲑虱(Lepeophtheirus salmonis salmonis)丰度的监测以及关键生活史特征的参数化,一直受到使用传统光学显微镜进行劳动密集型和易出错的量化的阻碍。荧光照明已被提议作为改善可视化的一种手段,但在此之前,尚未对相关荧光谱和测量条件进行充分的研究。我们使用激发和发射矩阵(200-600nm)研究了 L. salmonis 和非目标桡足类的荧光谱,并确定了独特的荧光信号。使用 470±40nm 的激发波长和 525±50nm 的发射波长的荧光显微镜显示,在福尔马林储存 90 天后,鲑虱的平均荧光强度比非目标桡足类(桡足幼体和成虫阶段)高 2.4 倍。在福尔马林中进行 7 天 42°C 的热处理,将鲑虱桡足幼体和非目标桡足类之间的差异增加到 3.6 倍,从而无需进行长时间的储存。我们还研究了荧光信号和内源性荧光团与海虱种类、年龄、阶段和宿主鱼来源的变化之间的关系。在本文所述的条件下,发现荧光信号是一种可靠的方法,可以将鲑虱与非目标浮游动物区分开来。荧光信号的适应性将大大加快浮游生物样本中鲑虱幼虫的传统计数方法,并提供一种自动检测的手段。

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