水域生态学中生物稳定同位素样品采集、处理与保存

徐军; 王玉玉; 王康; 曾庆飞; 张敏; 张欢

doi:10.7541/2020.114

水域生态学中生物稳定同位素样品采集、处理与保存

徐军^1, ,,
王玉玉²,
王康¹,
曾庆飞³,
张敏⁴,
张欢⁵

1.
中国科学院水生生物研究所, 武汉 430072
2.
北京林业大学生态与自然保护学院, 北京 100083
3.
中国科学院南京地理与湖泊研究所, 南京 210008
4.
华中农业大学水产学院, 武汉 430070
5.
南昌大学生命科学学院, 南昌 330031

基金项目: 国家重点研发计划项目(2018YFD0900904); 中国科学院国际合作重点项目(152342KYSB20190025)资助

详细信息

通信作者:
徐军(1978—), 男, 研究员, 博士生导师; 主要从事水域生态学与同位素生态学研究。E-mail: xujun@ihb.ac.cn

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出版历程
- 收稿日期: 2020-02-17
- 修回日期: 2020-07-20
- 网络出版日期: 2020-09-14
- 发布日期: 2020-09-29

PROTOCOLS FOR SAMPLE COLLECTION, PRETREATMENT AND PRESERVATION OF AQUATIC ORGANISMS IN STABLE ISOTOPE ECOLOGY

1.
Institute of Hydrobiology, Chinese Academy of Sciences , Wuhan 430072, China
2.
School of Ecology and Nature Conservation, Beijing Forestry University, Chinese Academy of Science, Beijing 100083, China
3.
Nanjing Institute of Geography and Limnology, Chinese Academy of Science, Nanjing 210008, China
4.
College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China
5.
School of Life Sciences, Nanchang University, Nanchang 330031, China

Funds: Supported by the National Key Research and Development Program of China (2018YFD0900904); the International Cooperation Project of the Chinese Academy of Sciences (152342KYSB20190025)

Corresponding author:

摘要

摘要: 稳定同位素分析技术由于能够刻画复杂的食物网结构并追踪食物网中的能量流而成为水域生态学研究中的重要手段。但是当水生生物样品采集、处理和保存过程中存在不确定性时, 营养关系分析中的同位素结果可能会产生误导性解释。文章采用数据模拟分析和文献总结的方法, 研究了水域生态系统中样品采集、处理和保存对于稳定同位素的影响, 概括性地建议了水域生态系统中适合应用稳定同位素分析技术开展生态学研究的样品采集、处理和保存的注意事项。但今后仍需进一步评估样品采集、处理和保存对稳定同位素比值的影响效果, 确定化学动力学在水生生物样品采集、处理和保存中的作用, 以进一步完善水生生物样品的采集、处理和保存稳定同位素生态学研究规范。
- 水域生态学 /
- 稳定同位素 /
- 营养生态位 /
- 基准 /
- 初级生产者 /
- 初级消费者 /
- 鱼类 /
- 分馏 /
- 周转 /
- 酸化 /
- 预处理 /
- 去脂 /
- 福尔马林 /
- 乙醇 /
- 腌渍
Abstract: Stable isotope studies are extremely useful for improving general descriptions of aquatic ecosystems due to their ability to simultaneously summarize complex trophic networks and track the energy flow through them. However, when considering that trophic relationships involve the uncertainty of sample collection, handling and preservation of aquatic organisms, application of general isotopic interpretations may easily lead to misleading conclusions. In these cases, an accepted protocol for the current process in the isotopic research should be considered. In this review, we first measured the uncertainty (isotopic shift) due to sample collection, handling and preservation in a system composed of one consumer and four potential food sources; we then showed four extreme cases to interpret the effect on the relative proportions of sources in the consumer’s diet due to uncertainty in a mixing-polygon model in Isosource. The effects of sample collection, handling and preservation of aquatic organisms on stable isotopes were briefly listed. All these methods had mixed results concerning their effect on the samples tested. For the sample collection, factors, such as, the uniformity, species composition, species life cycles, habitats, tissue difference and turnover, can affect the stable isotope composition. For sample handling, inorganic carbon, gut contents of small animals, and lipid content in the samples were considered as the common factors. For sample preservation, the effects of preservative, time, species or tissue were common on the stable isotope signals. Protocols for sample collection, handling and preservation of aquatic organisms in stable isotope ecology were then recommended. For sample collection, we advise using homogeneous and species-specific samples whenever possible, or performing complementary testing of the bulk- and tissue-specific sampling method. For sample handling, we advocate the gut evacuation for the samples using whole individual, acidification for the particulate organic samples, and lipid extraction and normalization for the samples with high lipid content. For sample preservation, we advocate using fresh material prepared immediately whenever possible, or using drying and freeze-drying when the immediate analysis is not possible, or conducting complementary testing of the preservative method for museum samples. In conclusion, we suggest suitable methods for sample collection, handling and preservation of aquatic organisms for ecological applications of isotopic analysis. The effects of sample collection, handling and preservation on stable isotope ratios need further evaluation. Research is also needed to determine the chemical dynamics in aquatic organisms responsible for the isotopic differences observed among different methods of sample collection, handling and preservation. We encourage further studies of method-specific isotopic variability to improve the normalization for sample collection, handling and preservation of aquatic organisms in stable isotope ecology.
- Aquatic ecology /
- Stable isotope /
- Trophic position /
- Baseline /
- Primary producer /
- Primary consumer /
- Fish /
- Fractionation /
- Turnover rate /
- Acidification /
- Pretreatment /
- Lipid extraction /
- Formalin /
- Ethanol /
- Salt

HTML全文

图 1 不确定性度量及消费者与四种食物来源的营养关系定义

a. 测试由于样品收集, 处理和保存引起的不确定性(同位素位移); Δ¹³C测量水平距离, Δ¹⁵N测量垂直距离, 不确定性(同位素位移)测量坐标空间中经解析的真值(T)和不确定性值(U)之间的直线距离; b. 由一个消费者及其四个潜在食物来源组成的明确的营养体系; 实线表示在IsoSource中定义为混合多边形的潜在源周围的凸包; 阴影区域表示不确定性, 圆圈数字表示潜在的食物来源, A、B、C和D表示四种极端情况, 以便于解释由于不确定性而导致的对消费者饮食中各种来源的相对比例的影响(图 2)

Figure 1. Measurement of uncertainty and the defined trophic system composed of one consumer and its four potential food sources

a. Measurement of uncertainty (isotopic shift) due to sample collection, handling and preservation. Δ¹³C measures the horizontal distance, Δ¹⁵N measures the vertical distance, and uncertainty (isotopic shift) measures the straight-line distance between the pared truth value (T) and uncertainty value (U) in coordinate space; b. A defined trophic system composed of one consumer and its four potential food sources. Solid lines show the convex hull around potential sources defined as mixing polygons in IsoSource. Shaded area represents the uncertainty, numbers in circles represent the potential food sources, and A, B, C, and D represent four extreme cases to facilitate interpretation of effect on the relative proportions of sources in the consumer diet resulted from uncertainty (Fig. 2)

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图 2 消费者四种食物来源不同情境下的分布频次特征

直方图表示从上述来源到消费者摄食中可行的IsoSource贡献分布。显示值的分布范围为1%—99%

Figure 2. Frequency of consumer’s four food sources in different contexts

The histograms provided the distribution of feasible IsoSource contributions from these sources to the consumer diet. The displayed values are the 1%—99% percentile ranges for these distributions

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图 3 建议在稳定同位素生态学中对水生生物样品收集、处理和保存的步骤图

Figure 3. Diagram of the logical steps suggested for sample collection, handling and preservation of aquatic organisms in stable isotope ecology

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水域生态学中生物稳定同位素样品采集、处理与保存

通信作者: 徐军(1978—), 男, 研究员, 博士生导师; 主要从事水域生态学与同位素生态学研究。E-mail: xujun@ihb.ac.cn

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出版历程

PROTOCOLS FOR SAMPLE COLLECTION, PRETREATMENT AND PRESERVATION OF AQUATIC ORGANISMS IN STABLE ISOTOPE ECOLOGY

Corresponding author:

计量

出版历程

目录

通信作者:
徐军(1978—), 男, 研究员, 博士生导师; 主要从事水域生态学与同位素生态学研究。E-mail: xujun@ihb.ac.cn