保存方法对大洋性头足类多组织稳定同位素分析的影响：以茎柔鱼为例

沈永富; 耿亮; 贡艺; 丁宇; 李云凯; 邓江山; 陈浩楠

doi:10.12131/20230091

保存方法对大洋性头足类多组织稳定同位素分析的影响：以茎柔鱼为例

doi: 10.12131/20230091

沈永富^1,,
耿亮¹,
贡艺^{1, 2, 3, 4, ,},
丁宇¹,
李云凯^{1, 2, 3, 4},
邓江山¹,
陈浩楠¹

1.
上海海洋大学海洋科学学院，上海 201306
2.
大洋渔业资源可持续开发教育部重点实验室，上海 201306
3.
国家远洋渔业工程技术研究中心，上海 201306
4.
农业农村部大洋渔业开发重点实验室，上海 201306

基金项目: 国家自然科学基金面上项目 (31900333, 42276092)；农业农村部公海渔业资源综合科学调查专项

详细信息

作者简介:
沈永富 (1997—)，男，博士研究生，研究方向为摄食生态学。E-mail: 1016959821@qq.com

通讯作者:
贡　艺 (1990—)，男，讲师，博士，研究方向为大洋生物栖息环境与生态学。E-mail: ygong@shou.edu.cn

中图分类号: Q 178
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- 文章访问数: 50
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- 被引次数: 0
出版历程
- 收稿日期: 2023-04-28
- 修回日期: 2023-05-24
- 录用日期: 2023-06-05
- 网络出版日期: 2023-08-16
- 刊出日期: 2023-10-05

Effects of preservation methodology on multi-tissue stable isotope signatures of oceanic squid: a case study of jumbo squid (Dosidicus gigas)

SHEN Yongfu^1
,,
GENG Liang¹,
GONG Yi^{1, 2, 3, 4
, ,},
DING Yu¹,
LI Yunkai^{1, 2, 3, 4},
DENG Jiangshan¹,
CHEN Haonan¹

1.
College of Marine Sciences, Shanghai Ocean University, Shanghai 201306, China
2.
Key Laboratory of Sustainable Exploitation of Oceanic Fisheries Resources, Ministry of Education, Shanghai 201306, China
3.
National Engineering Research Center for Oceanic Fisheries, Shanghai 201306, China
4.
Key Laboratory of Oceanic Fisheries Exploration, Ministry of Agriculture and Rural Affairs, Shanghai 201306, China

摘要

摘要: 碳、氮稳定同位素技术 (δ¹³C和δ¹⁵N) 是海洋动物摄食生态学研究的主要方法之一。目前对海洋动物各组织的保存方法尚无统一的标准，这可能造成测定结果出现偏差，并产生误导性结论。以大洋性头足类茎柔鱼 (Dosidicus gigas) 为研究对象，开展了冷冻保存 (−20 ℃)、乙醇 (体积分数75%) 与甲醛 (体积分数38%) 溶液保存对肌肉、角质颚和性腺组织δ¹³C、δ¹⁵N和碳氮比 (C/N) 测定的比较研究。结果表明，冷冻保存在第30天时仅对肌肉和性腺组织δ¹³C和C/N值产生了显著性影响 (P<0.05)，偏移量分别为(−0.89±0.30)‰和0.22±0.15，但相比于乙醇和甲醛方法偏移量较小。乙醇保存第30天时，肌肉δ¹³C、δ¹⁵N和C/N值均有显著性变化 (P<0.05)，偏移量分别为(−0.41±0.50)‰、(0.76±0.79)‰和0.34±0.05；性腺δ¹³C和和C/N值存在显著性变化 (P<0.05)，偏移量分别为 (−0.36±0.44)‰和 0.21±0.14；而角质颚的变化不显著 (P>0.05)。甲醛保存会导致3种组织的δ¹³C、δ¹⁵N和C/N值出现显著性变化 (P<0.05)，30 d后 δ¹³C 、δ¹⁵N和 C/N值总偏移量分别为 (−3.03±1.87)‰、(−0.48±0.72)‰和0.86±0.73，但保存至第180天时仅角质颚和性腺的δ¹⁵N和C/N值仍持续增大 (P<0.05)，可能与组织特异性有关。因此在分析稳定同位素时，应谨慎使用长时间保存后的软组织样品 (肌肉和性腺)，而硬组织 (角质颚) 可采用冷冻或乙醇保存。
- 茎柔鱼 /
- 稳定同位素 /
- 冷冻 /
- 乙醇 /
- 甲醛
Abstract: Carbon and nitrogen stable isotope technology (δ¹³C and δ¹⁵N) is an important tool for the study of trophic ecology of marine organisms. However, there is no unified standard for the preservation methods in cephalopods' samples, which may cause biases in stable isotope values and easily lead to misleading conclusions. Thus, we assessed the effects of preservation method (Freezing, 75% ethanol solution and 38% formaldehyde solution) and duration (0, 30 and 180 d) on the stable isotope ratios of carbon and nitrogen of tissues (Muscle, beak and gonad) of jumbo squid (Dosidicus gigas). The results show that freezing affected δ¹³C and C/N values of muscle and gonad within 30 d (P<0.05), with offsets of (−0.89±0.30)‰ and 0.22±0.15, respectively, which were lower than the effects of alcohol and formaldehyde. After 30 days of storage in the alcohol solution, significant changes were observed for muscle [δ¹³C: (−0.24±0.42)‰ , δ¹⁵N: (0.17±1.07)‰ and C/N: (0.32±0.49)] and gonad [δ¹³C: (−0.36±0.44), C/N: (0.21±0.14)] (P<0.05), but no significant change was found in beak (P>0.05). Formaldehyde preservation induced a decrease in δ¹³C [(−3.03±1.87)‰] and δ¹⁵N [(−0.48±0.72)‰] but an increase in C/N (0.86±0.73) within 30 d (P<0.05). However, the δ¹⁵N and C/N values significantly increased in beak and gonad stored in formaldehyde on the 180^th day (P<0.05). Therefore, it should be careful to use the preserved soft tissue samples during stable isotope analysis, while hard tissue, such as horny jaw, can be preserved by freezing or ethanol solution.
- Dosidicus gigas /
- Stable isotope /
- Freezing /
- Ethanol /
- Formaldehyde

HTML全文

图 1 3种保存方法对茎柔鱼肌肉、角质颚和性腺组织δ¹³C、δ¹⁵N 和 C/N 的影响

Figure 1. Effect of three treatments on δ¹³C and δ¹⁵N and C/N of D. gigas muscle, beak and gonad tissues

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表 1 茎柔鱼 3 种组织不同保存方式下的稳定同位素比值

Table 1. δ¹³C, δ¹⁵N values and C/N ratios of different preservation methods in three tissues of D. gigas

	组织 Tissue	对照 Control	−20 ℃ 冷冻 Freezing		75% 乙醇 Ethanol		38% 甲醛 Formaldehyde
	组织 Tissue	第0天 Day 0	第30天 Day 30	第180天 Day 180	第30天 Day 30	第180天 Day 180	第30天 Day 30	第180天 Day 180
肌肉 Muscle	δ¹³C	−17.97±0.30^a	−18.73±0.26^b	−19.31±0.33^b	−18.40±0.56^b	−18.47±0.37^b	−22.36±3.61^b	−21.66±1.31^b
	δ¹⁵N	15.76±2.73^a	16.06±2.80^a	15.94±3.06^a	16.47±2.58^b	16.62±2.71^b	14.95±2.17^b	16.09±2.60^b
	C/N	3.22±0.07^a	3.49±0.06^b	3.51±0.12^b	3.57±0.14^b	4.25±0.96^c	4.79±1.15^b	4.66±1.56^b
角质颚 Beak	δ¹³C	−18.48±0.48^a	−18.54±0.53^a	−18.47±0.51^a	−18.50±0.52^a	−18.51±0.52^a	−20.50±2.29^b	−19.96±1.52^b
	δ¹⁵N	10.61±0.98^a	10.58±1.10^a	10.25±1.14^a	10.61±1.17^a	11.39±1.02^a	9.85±1.04^b	10.82±0.61^a
	C/N	5.38±0.51^a	5.56±0.32^a	5.52±0.52^a	5.47±0.53^a	5.46±0.57^a	5.81±0.52^b	6.36±0.40^c
性腺 Gonad	δ¹³C	−17.61±0.79^a	−18.62±0.87^b	−18.94±0.56^b	−17.97±0.58^b	−18.94±0.56^c	−21.56±0.85^b	−21.40±0.80^b
	δ¹⁵N	14.88±0.96^a	14.82±1.03^a	14.22±1.04^a	14.92±1.02^a	14.65±1.01^a	14.68±1.04^b	15.02±1.17^c
	C/N	3.21±0.16^a	3.57±0.33^b	3.55±0.35^b	3.42±0.13^b	3.91±0.22^c	3.97±0.36^b	3.93±0.37^c
注：同行数据不同上标字母表示有显著性差异 (P<0.05)。 Note: Values in each row followed by different superscript letters were significant differences (P<0.05).

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