Effect evaluation of alizarin complexone marking for otoliths of juvenile black carp (Mylopharyngodon piceus)
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摘要: 放流标记技术研究是支撑增殖放流效果评估的重要内容。该研究用质量浓度为100 mg·L−1的茜素络合物 (ALC) 对约2月龄的青鱼 (Mylopharyngodon piceus) 幼鱼进行48 h浸泡，随后移入正常水体中恢复续养20 d。对其3种耳石在可见光和不同的激发光下进行比较分析，评价了茜素络合物在不同耳石上的沉积情况与标记效果。结果表明，通过在可见光和不同的激发光下进行观察，其3种耳石上均可见明显的标记区，标记率达100%。在对标记鱼恢复续养20 d后发现，耳石上的荧光标记区强度仍很高，表现出了较好的标记效果。此外，相关标记的最佳检测光源为蓝色激发光，绿色激发光次之。微耳石是用于青鱼幼鱼耳石荧光标记观察的最适类型，同时也是用于时滞检测的最适类型，其次是星耳石。此外，将鱼体浸泡入茜素络合物溶液中至其在耳石上形成标记沉积，以及将鱼体移出浸泡溶液至其耳石上标记区域消失，均存在1 d时滞。Abstract: Investigation on artificial marking technology is very important supporting work for national fish mark release recapture and effectiveness tracing evaluation issues. In this study, two-month old juvenile black carps (Mylopharyngodon piceus) had been immersed in alizarin complexone (ALC) at a concentration of 100 mg·L−1 for 48 h, and then transferred to normal water for 20 d. Three pairs of otoliths (i.e., sagitta, lapillus and asteriscus) were removed from the black carps by continuous sampling, observed under fluorescence microscope and photographed, so as to evaluate the characteristics of deposition and the marking effect of ALC in different otoliths. The results show that the marked areas were clearly observed in each otolith pair under visible light and different excitation light, with a 100% mark success rate. After 20 d of recovery culture in normal water without ALC, the intensity of ALC mark in otoliths of the marked fish was still very high, showing a good marking effect. Moreover, the optimum light source was blue excitation light, followed by green excitation light. The lapillus was the optimal otolith type for ALC marking and for judgment of time lag among the three pairs of otoliths, followed by asteriscus. In addition, there was a time lag of 1 day between the carps immersed in ALC solution and ALC deposition into otolith, as well as between removing immersed carps from ALC solution and disappearance of ALC mark from otolith.
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Key words:
- Mylopharyngodon piceus /
- Otolith /
- Fluorescent marking /
- Alizarin complexone /
- Enhancement release
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图 1 青鱼幼鱼的3类耳石
a. 矢耳石；b. 微耳石；c. 星耳石。
Figure 1. Three types of otoliths from juvenile black carp
a. Sagitta; b. Lapillus; c. Asteriscus.
图 2 不同取样天数标记组和对照组青鱼幼鱼的体质量和全长变化 (N=5)
*. 两组间差异显著 (P<0.05)。
Figure 2. Change of body mass and total length of black carp of marking group and control group on different sampling day (N=5)
*. Significant difference between two groups (P<0.05).
图 3 经茜素络合物标记后恢复续养第10天时青鱼幼鱼矢耳石、微耳石、星耳石分别在可见光、蓝色激发光、绿色激发光和紫色激发光下的标记效果
a1. 矢耳石分别在可见光、蓝色激发光、绿色激发光和紫色激发光下的标记效果；b1. 微耳石分别在可见光、蓝色激发光、绿色激发光和紫色激发光下的标记效果；c1. 星耳石分别在可见光、蓝色激发光、绿色激发光和紫色激发光下的标记效果；图中白色箭头表示由矢耳石、微耳石、星耳石核心指向荧光标记轮。
Figure 3. Marking effect of three types of otoliths of juvenile black carp under different light sources on 10th day of recovery culture after ALC immersion
a1. Marking effect of sagitta under visible light, blue excitation light, green excitation light and purple excitation light; b1. Marking effect of lapillus under visible light, blue excitation light, green excitation light and purple excitation light; c1. Marking effect of asteriscus under visible light, blue excitation light, green excitation light and purple excitation light. The white arrows show that the core of sagitta, lapillus and asteriscus point to the fluorescent marker ring.
图 4 青鱼幼鱼经茜素络合物浸泡标记后恢复续养第0、第5、第10、第15和第20天时微耳石在蓝色激发光的标记效果
Figure 4. Marking effect of lapillus of juvenile black carp under blue light source on 0th, 5th, 10th, 15th, 20th day of recovery culture after ALC immersion
图 5 青鱼幼鱼经茜素络合物浸泡标记后恢复续养第5天时微耳石标记效果
2个白色箭头分别指向微耳石的2条紫红色标记环；黑色短线表示微耳石新形成的日轮所在位置，即为一明带和一暗带组合；4个黑色箭头则分别指向微耳石外部新形成的4个日轮。
Figure 5. Marking effect of lapillus of juvenile black carp under four light sources on 5th day of recovery culture after ALC immersion
Two white arrows point to two purple red marker rings in lapillus; the black dash shows the position of the newly formed daily growth increments in lapillus, which is a combination of a bright band and a dark band; the four black arrows point outward to the four newly formed daily growth increments in lapillus.
表 1 青鱼幼鱼3类耳石在不同光源下的检测效果
Table 1. Detection effect of three types of otoliths from juvenile black carp under four different light sources
恢复续养天数
Days of recovery
culture/d实验鱼数
Number of
fish耳石类型
Otolith
type耳石标本数
Number of
otolith标记强度 Marker intensity 可见光
BS蓝色激发光
WBS绿色激发光
WGS紫色激发光
WUS0 5 矢耳石 10 2 (++) 2 (++) 2 (++) 1 (+) 5 5 矢耳石 10 2 (++) 3 (+++) 3 (+++) 2 (++) 10 5 矢耳石 10 2 (++) 4 (++++) 3 (+++) 2 (++) 15 5 矢耳石 10 1 (+) 4 (++++) 3 (+++) 2 (++) 20 5 矢耳石 10 1 (+) 3 (+++) 2 (++) 1 (+) 0 5 微耳石 10 2 (++) 3 (+++) 3 (+++) 2 (++) 5 5 微耳石 10 3 (+++) 4 (++++) 4 (++++) 3 (+++) 10 5 微耳石 10 3 (+++) 4 (++++) 4 (++++) 3 (+++) 15 5 微耳石 10 2 (++) 4 (++++) 4 (++++) 3 (+++) 20 5 微耳石 10 2 (++) 4 (++++) 4 (++++) 2 (++) 0 5 星耳石 10 2 (++) 2 (++) 2 (++) 1 (+) 5 5 星耳石 10 3 (+++) 3 (+++) 2 (++) 1 (+) 10 5 星耳石 10 3 (+++) 3 (+++) 2 (++) 1 (+) 15 5 星耳石 10 3 (+++) 2 (++) 1 (+) 1 (+) 20 5 星耳石 10 2 (++) 2 (++) 1 (+) 1 (+) 
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