福建平潭海域不同年龄仿刺参骨片种类和形态学研究

    Study on morphology and species of ossicles of different ages of sea cucumber (Apostichopus japonicus) collected from Pingtan sea area, Fujian Province

    • 摘要:
      目的 海参真皮的表层包含有被称为骨片的内骨骼,是海参纲最重要的特征。利用不同类型的海参骨片及其相对比例能够对不同种类的海参予以快速鉴定。
      方法 本文采用NaClO法、蛋白酶K法,对福建平潭1、2、3龄仿刺参(Apostichopus japonicus)的各组织(棘刺、体壁、触手、管足、纵肌、呼吸树、肠道)进行采集与处理,得到不同年龄仿刺参不同组织骨片类型,并分别比较采用NaClO法、蛋白酶K法制备的仿刺参骨片的显微图像,利用SPSS软件统计不同仿刺参骨片类型的相对比例。
      结果 在显微镜和扫描电子显微镜下能观察到7种仿刺参骨片类型,包括5种主要类型(桌形体、扣形体、杆状体、长孔状体、花纹状体)和2种特殊类型(复合盘状骨片和C形体),且不同组织内的仿刺参骨片类型存在显著性差异(P<0.05)。在对比NaClO法、蛋白酶K法制得的仿刺参骨片图像时,发现2种方法均可得到仿刺参骨片,其中经NaClO法析出的仿刺参骨片能被较清楚地观察到骨片种类与大致的数量,且不附带其他组织,标本干净,但骨片因过脆、易损坏而结构不完整;蛋白酶K法得到的仿刺参骨片同样能与其他组织分离,且具有一定的韧性,在显微镜下其结构更加完整,能被更好地计数与统计。通过统计仿刺参不同年龄段各组织内骨片类型的相对比例,发现不同年龄不同组织仿刺参骨片类型相对比例存在显著性差异(P<0.05)。1龄仿刺参的体壁、棘刺、管足和触手均含有扣形体、桌形体、长孔状体,其中扣形体在体壁中的相对比例最高,达到83.28%±9.33%;桌形体的相对比例在管足中最高,达到35.02%±10.84%;长孔状体在触手中的相对比例较高,达到7.23%±2.50%。1龄仿刺参体壁和管足中桌形体的相对比例分别为20.28%±7.29%和35.02%±10.84%,显著高于2龄仿刺参(3.80%±1.35%、17.21%±1.87%)和3龄仿刺参(3.40%±1.43%、11.23%±1.01%)(P<0.05)。
      结论 不同年龄仿刺参的不同组织骨片的结构类型不一,相对比例不同,且能通过多种方法获得,具有稳定性,是鉴别海参种属的重要指标。
      意义 本研究为仿刺参生物学鉴定提供基础资料,为其年龄鉴别提供借鉴。

       

      Abstract:
      Background At present, domestic and foreign scholars have used morphology, spectroscopy, mass spectrometry, molecular biology and other technologies to identify sea cucumber from the morphology, chemical substances contained, nucleic acid differences and other aspects of sea cucumber, but considering the technical requirements and time cost, morphology is still the main identification method of sea cucumber.
      Objective The dermis surface layer of sea cucumbers contains ossicles, which constitute the distinguishing feature of this class. The types and relative proportions of these ossicles can facilitate rapid identification of different sea cucumber species.
      Methods In this study, NaClO and protease K methods were employed to extract and process tissues of sea cucumbers (Apostichopus japonicus) aged 1 to 3 years from the Pingtan sea area (Fujian, China). Tissue samples included spines, body walls, tentacles, tube feet, longitudinal muscles, respiratory trees, and intestines. Ossicle images obtained through these methods were compared, and the relative proportions of each type of ossicle were analyzed using SPSS software.
      Results The results revealed seven types of ossicles: table-shaped, button-shaped, rod-shaped, rosette-shaped, long hole-like body ossicles, composite discoid ossicles, and C-shaped ossicles, observable under both light and electron microscopes, with significant differences (P<0.05). Comparing the images obtained by NaClO and protease K methods, NaClO-extracted ossicles displayed clearer types and approximate numbers, free from tissue attachment, but were brittle and prone to damage, resulting in incomplete structures. In contrast, protease K-extracted ossicles showed better structural integrity under the microscope, facilitating more accurate counting and analysis, although they retained some tissue. Additionally, the relative proportions of ossicle types significantly differed (P<0.05) across different ages and tissues. For instance, in one-year-old specimens, button-shaped ossicles constituted the highest proportion in the body wall 83.28%±9.33%, while table-shaped ossicles dominated in the body cavity 35.02%±10.84%, and long hole-like body ossicles were most prevalent in tentacles 7.23%±2.50%. Relative proportions in the body wall and tube feet at one year (20.28%±7.29% and 35.02%±10.84%, respectively) were significantly higher than in the second year (3.80%±1.35% and 17.21%±1.87%, respectively) and third year (3.40%±1.43% and 11.23%±1.01%, respectively) (P<0.05).
      Conclusion The results show that the relative proportion of ossicle types of sea cucumber have a difference in different ages and tissues, and can be obtained by various methods stability, which is an important index to identify sea cucumber species.
      Significance This study provides the basic data for the biological identification of the A. japonicus and the reference for the age identification.

       

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