将视觉和分子方法结合起来进行鱼卵分析:甲醛固定和储存程序研究

IF 2.1 3区 地球科学 Q2 LIMNOLOGY Limnology and Oceanography: Methods Pub Date : 2024-08-02 DOI:10.1002/lom3.10640
Bahar Mozfar, Anders Thorsen, Alejandro Mateos‐Rivera, Geir Dahle, Rasmus Skern‐Mauritzen, Henning Wehde, Bjørn A. Krafft
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引用次数: 0

摘要

鱼卵的准确分类和发育阶段测定对于生态监测、保护工作和种群评估至关重要。鱼类和渔业的传统方法依赖于目测形态特征,但由于物种重叠,尤其是早期阶段的物种重叠,这些方法面临挑战。DNA 条形码等分子工具可提供更高分辨率的分类鉴定,但可能无法提供发育阶段的信息。本研究探讨了不同甲醛固定浓度和储存程序对挪威罗弗敦岛收集的鱼卵进行视觉和分子分析的有效性。视觉分析成功地确定了所有固定溶液的发育阶段。使用 16S rRNA 基因进行分子条形码鉴定,100% 的鱼卵都能鉴定出鱼种,在甲醛固定条件下储存的成功率随时间推移而降低。将卵转移到乙醇中后,用 4% 的甲醛固定 12 或 24 小时,DNA 条形码的成功率最高。使用 0.5%和 1%甲醛固定长达 8 周也能获得较高的 DNA 成功率,但随着储存时间的延长,结果会越来越差。这项研究为整合鱼卵分析的视觉和分子方法提供了宝贵的见解,对海洋调查期间的样本保存具有实际意义。
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Integrating visual and molecular approaches for fish eggs analysis: A study on formaldehyde fixation and storage procedures
Accurate taxonomic classification and developmental stage determination of fish eggs are crucial for ecological monitoring, conservation efforts, and stock assessments. Traditional methods for fish and fisheries rely on visual examination of morphological traits, but they face challenges due to species overlap especially for early stages. Molecular tools, such as DNA barcoding, offer higher resolution in taxonomic identification but may not provide developmental stage information. This study explores the effectiveness of different formaldehyde fixation concentrations and storage procedures on fish eggs collected from Lofoten, Norway, for both visual and molecular analysis. Visual analysis successfully identified developmental stage for all fixation solutions. Molecular barcoding using the 16S rRNA gene identified up to 100% of eggs at the species level, with decreasing success rates over time when stored in formaldehyde fixation. The highest DNA barcoding success rates were accomplished using 4% formaldehyde fixation for 12‐ or 24‐h following transfer to ethanol. Using 0.5% and 1% formaldehyde fixation up to 8 weeks also resulted in high DNA success rates, but results deteriorated with increasing storage time. This study provides valuable insights for integrating visual and molecular methods for fish egg analysis, with practical implications for sample preservation during marine surveys.
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来源期刊
CiteScore
4.80
自引率
3.70%
发文量
56
审稿时长
3 months
期刊介绍: Limnology and Oceanography: Methods (ISSN 1541-5856) is a companion to ASLO''s top-rated journal Limnology and Oceanography, and articles are held to the same high standards. In order to provide the most rapid publication consistent with high standards, Limnology and Oceanography: Methods appears in electronic format only, and the entire submission and review system is online. Articles are posted as soon as they are accepted and formatted for publication. Limnology and Oceanography: Methods will consider manuscripts whose primary focus is methodological, and that deal with problems in the aquatic sciences. Manuscripts may present new measurement equipment, techniques for analyzing observations or samples, methods for understanding and interpreting information, analyses of metadata to examine the effectiveness of approaches, invited and contributed reviews and syntheses, and techniques for communicating and teaching in the aquatic sciences.
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Issue Information Issue Information Correction to “Estimating ethanol correction factors for δ13C and δ15N isotopic signatures of freshwater zooplankton from multiple lakes” Multivariate statistical “unmixing” of Indian and Pacific Ocean sediment provenance Assessment of a pH optode for oceanographic moored and profiling applications
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