CN114793988B - A method for identifying fertilized eggs of ray ray based on morphology - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 13
- 235000013601 eggs Nutrition 0.000 title claims abstract description 8
- 210000003278 egg shell Anatomy 0.000 claims abstract description 87
- 102000002322 Egg Proteins Human genes 0.000 claims abstract description 70
- 108010000912 Egg Proteins Proteins 0.000 claims abstract description 70
- 241000251468 Actinopterygii Species 0.000 claims abstract description 18
- 230000000877 morphologic effect Effects 0.000 claims abstract description 18
- 238000005259 measurement Methods 0.000 claims abstract description 12
- 238000012216 screening Methods 0.000 claims abstract description 4
- 230000000241 respiratory effect Effects 0.000 claims description 4
- 210000004681 ovum Anatomy 0.000 claims 1
- 241000251730 Chondrichthyes Species 0.000 abstract description 8
- 241000039233 Okamejei kenojei Species 0.000 abstract description 5
- 238000009395 breeding Methods 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract description 4
- 238000011835 investigation Methods 0.000 abstract description 4
- 238000009472 formulation Methods 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- 238000012544 monitoring process Methods 0.000 abstract description 2
- 241001670157 Gymnura Species 0.000 description 3
- 230000000366 juvenile effect Effects 0.000 description 3
- 241000894007 species Species 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000251477 Chimaera Species 0.000 description 1
- 229920002567 Chondroitin Polymers 0.000 description 1
- 241000238424 Crustacea Species 0.000 description 1
- 241000229987 Okamejei Species 0.000 description 1
- 241001415775 Rajidae Species 0.000 description 1
- 241000251184 Rajiformes Species 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- DLGJWSVWTWEWBJ-HGGSSLSASA-N chondroitin Chemical compound CC(O)=N[C@@H]1[C@H](O)O[C@H](CO)[C@H](O)[C@@H]1OC1[C@H](O)[C@H](O)C=C(C(O)=O)O1 DLGJWSVWTWEWBJ-HGGSSLSASA-N 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000002224 dissection Methods 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
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- 238000005070 sampling Methods 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
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- A01K61/00—Culture of aquatic animals
- A01K61/90—Sorting, grading, counting or marking live aquatic animals, e.g. sex determination
- A01K61/95—Sorting, grading, counting or marking live aquatic animals, e.g. sex determination specially adapted for fish
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Abstract
Description
技术领域technical field
本发明属于海洋生物繁育中的软骨鱼类仔稚幼鱼鉴定与监测技术领域,具体涉及一种基于形态学鉴定孔鳐受精卵的方法。The invention belongs to the technical field of identification and monitoring of juvenile cartilaginous fish larvae in the breeding of marine organisms, and in particular relates to a method for identifying fertilized eggs of ray rays based on morphology.
背景技术Background technique
孔鳐(Okamejei kenojei Müller&Henle 1841),俗称老板鱼,隶属软骨鱼纲(Chondrichthyes)、鳐目(Rajiformes)、鳐科(Rajidae)、瓮鳐属 (Okamejei),俗称老板鱼,主要分布于环太平洋西岸的海域,在我国主要分布于黄渤海等海域。属冷水性底层鱼类,主要以底栖甲壳类等生物为食。孔鳐是我国北方沿海地区常见的经济鱼类。同时由于其体内富含软骨素等成分,使其兼具药用价值;再加之其游动时的独特体态,使其也成为重要的水族观赏鱼类。Okamejei kenojei Müller&Henle 1841, commonly known as boss fish, belongs to Chondrichthyes, Rajiformes, Rajidae, Okamejei, commonly known as boss fish, mainly distributed in the west coast of the Pacific Rim In my country, it is mainly distributed in the Yellow Sea and Bohai Sea. It is a cold-water bottom fish and mainly feeds on bottom-dwelling crustaceans and other organisms. Kong ray is a common economic fish in the coastal areas of northern my country. At the same time, because its body is rich in chondroitin and other ingredients, it has both medicinal value; coupled with its unique posture when swimming, it also becomes an important aquarium ornamental fish.
相较于硬骨鱼类,软骨鱼类具有体型较大,繁殖周期长,幼鱼发育缓慢等特点,使得软骨鱼类在面临栖息环境的变化时,往往会出现巨大的种群波动,在过去的五十年间,全球软骨鱼类的数量急剧减少,部分种群甚至濒临灭绝。孔鳐的野生种群也面临着种质退化的风险,对于孔鳐的繁殖保育工作的开展日益迫切。如何有效地鉴别孔鳐野生种群的仔稚幼鱼,同时又避免解剖取样等操作对鱼体造成伤害,成为孔鳐繁殖保育的重要研究内容之一。Compared with bony fishes, cartilaginous fishes have the characteristics of larger body size, longer reproductive cycle, and slow growth of juveniles, which make cartilaginous fishes often experience huge population fluctuations when faced with changes in their habitats. In the past ten years, the number of cartilaginous fishes in the world has decreased sharply, and some populations are even on the verge of extinction. The wild population of Kong Ray is also facing the risk of germplasm degradation, and it is increasingly urgent to carry out the breeding and conservation of Kong Ray. How to effectively identify the larvae and juveniles of the wild population of the ray, and at the same time avoid the damage to the fish body caused by operations such as dissection and sampling, has become one of the important research contents for the reproduction and conservation of the ray.
相较于鲨类,鳐类的卵壳形态较为单一且相似,这使得鳐类卵壳形态的辨识中容易出现混淆等问题。虽然鳐鱼所产生的卵壳形态具有物种特异性的数值差异,且目前关于孔鳐卵壳形态学特征已有初步研究,但是相关指征较少,难以区分鳐类物种间差异。同时由于样品受限,目前鳐类卵壳形态学鉴定的定量指标还十分有限,多数研究主要以定性分析为主,这也为鳐鱼卵壳形态学比较与鉴定造成了诸多不便。此外,基于卵壳形态学鉴定软骨鱼类仔稚幼鱼的方法,在国内还尚未见报道。Compared with sharks, the eggshell morphology of rays is relatively single and similar, which makes the identification of the eggshell shape of rays prone to confusion and other problems. Although there are species-specific numerical differences in the eggshell morphology produced by rays, and there have been preliminary studies on the morphological characteristics of the eggshell of the ray, but there are few relevant indications, and it is difficult to distinguish the differences between the rays. At the same time, due to limited samples, quantitative indicators for the morphological identification of skate eggshells are still very limited, and most studies are mainly based on qualitative analysis, which also causes a lot of inconvenience for the comparison and identification of skate eggshell morphology. In addition, the method of identifying juvenile cartilaginous fish larvae based on eggshell morphology has not been reported in China.
发明内容Contents of the invention
本发明目的在于提供了一种简单,快捷基于形态学鉴定孔鳐受精卵的方法。The purpose of the present invention is to provide a simple and fast method for identifying fertilized eggs of ray ray based on morphology.
为实现上述目的,本发明采用技术方案为:In order to achieve the above object, the technical solution adopted by the present invention is:
一种基于形态学鉴定孔鳐受精卵的方法,取待鉴定软骨鱼类卵壳通过形态学观察初步筛选鳐鱼卵壳,对初步筛选为鳐鱼卵壳的样品的特征部位进行形态学测量,测定后各样品的特征部位的测量结果与孔鳐卵壳参考标准进行对比,各特征部位测量值与参考标准吻合,鉴定识别该待测样品为孔鳐所产卵壳。A method for identifying the fertilized eggs of ray ray based on morphology. The egg shells of cartilaginous fishes to be identified are initially screened for ray shells through morphological observation, and the characteristic parts of the samples that are initially screened as ray shells are morphologically measured. After the determination, the measurement results of the characteristic parts of each sample were compared with the reference standard of the ray shell.
所述形态学观察初步筛选鳐鱼卵壳为卵壳中呈左右对称中央隆起的袋状结构,壳体两侧加厚形成龙骨缘,龙骨缘沿壳体长轴分别向上和向下延伸形成四个角,四个角中末段沿卵壳壳体的隆起方向延伸,每个角的末端各有一呼吸裂,同时,同一侧的两个角之间通过裙边相连,即初筛为鳐类卵壳。According to the morphological observation, the ray shell is initially screened as a bag-like structure with left-right symmetrical central uplift in the egg shell. The two sides of the shell are thickened to form a keel edge, and the keel edge extends upward and downward along the long axis of the shell to form four The end of the four corners extends along the bulging direction of the egg shell, and each corner has a respiratory slit at the end. At the same time, the two corners on the same side are connected by a skirt, that is, the first screening is a ray egg shell.
所述孔鳐卵壳形态学特征部位为总长度、极宽、体长、前角长、后角长、前裙边长、后裙边长、龙骨缘宽度和附着丝距离。The morphological characteristic parts of the ray shell are total length, extreme width, body length, front horn length, rear horn length, front skirt length, rear skirt length, keel edge width and attachment wire distance.
所述各特征部位的测量:The measurement of each feature position:
总长度(TL):卵壳前角顶端与后角顶端之间的垂直距离;Total length (TL): the vertical distance between the top of the anterior horn of the eggshell and the top of the rear horn;
极宽(MW):卵壳左侧边缘与右侧边缘之间的最大水平距离;Extreme width (MW): the maximum horizontal distance between the left and right edges of the eggshell;
体长(BL):卵壳前裙边与后裙边之间的垂直距离;Body length (BL): the vertical distance between the front skirt and the back skirt of the eggshell;
前角长(AHL):卵壳前角顶端与前裙边外缘的垂直距离;Front horn length (AHL): the vertical distance between the top of the front corner of the eggshell and the outer edge of the front skirt;
后角长(PHL):卵壳后角顶端与后裙边外缘的垂直距离;Back horn length (PHL): the vertical distance between the top of the back horn of the eggshell and the outer edge of the back skirt;
前裙边长(AAL):卵壳前裙边的外缘与内缘之间的垂直距离;Front skirt length (AAL): the vertical distance between the outer edge and the inner edge of the eggshell front skirt;
后裙边长(PAL):卵壳后裙边的外缘与内缘之间的垂直距离;Posterior skirt length (PAL): the vertical distance between the outer edge and the inner edge of the rear skirt of the eggshell;
龙骨缘宽度(LKW):卵壳龙骨缘的外缘与内缘的水平距离;Keel width (LKW): the horizontal distance between the outer and inner edges of the eggshell keel;
附着丝距离(AFL):卵壳附着丝第一附着点和第三附着点之间的垂直距离。Attachment filament distance (AFL): the vertical distance between the first attachment point and the third attachment point of the eggshell attachment filament.
本发明所具有的优点:The advantages that the present invention has:
本发明基于孔鳐卵壳形态学指标,建立了孔鳐受精卵卵壳的鉴别标准,可以快速地鉴别孔鳐的受精卵,适用于野外调查中简易环境下评估孔鳐的种群,特别适用于非繁殖季节(孔鳐仔稚幼鱼已经孵化,卵壳中无活体样本可用的条件)进行的野外调查。该发明对于孔鳐种群的评估以及纯种种质的保育有一定参考价值。The present invention is based on the morphological index of the ray shell, establishes the identification standard of the fertilized egg shell of the ray, can quickly identify the fertilized eggs of the ray, is suitable for evaluating the population of the ray in a simple environment in field investigations, and is especially suitable for Field surveys conducted during the non-breeding season (conditions in which juveniles of ray ray have hatched and no live samples are available in the egg shells). The invention has certain reference value for the evaluation of the hole ray population and the conservation of pure germplasm.
附图说明Description of drawings
图1为孔鳐卵壳实物照片。Figure 1 is a real photo of the egg shell of the ray.
图2为孔瑶卵壳的测量标准参考图。Figure 2 is a standard reference map for the measurement of Kongyao eggshells.
图3为孔鳐卵壳的模式图。Fig. 3 is a schematic diagram of the egg shell of the ray.
具体实施方式detailed description
下面结合附图和具体实施例对本发明作进一步详细的说明。The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.
本发明基于孔鳐卵壳形态学建立鉴定孔鳐物种的方法,该方法能有效地根据形态学指标从卵壳样品中区分出孔鳐物种,适用于出海调查等简易实验条件;该方法对软骨鱼类针对性保育措施的制定具有一定参考价值。The invention establishes a method for identifying the species of the stingray based on the eggshell morphology of the stingray, and the method can effectively distinguish the stingray species from the eggshell sample according to the morphological index, and is suitable for simple experimental conditions such as going to sea for investigation; the method is effective for cartilage The formulation of targeted fish conservation measures has certain reference value.
实施例1Example 1
孔鳐卵壳形态学特征分为总长度、极宽、体长、前角长、后角长、前裙边长、后裙边长、龙骨缘宽度和附着丝距离,根据Treloar等人的相关研究结果获得孔鳐卵壳各特征的参考标准(Treloar等.,2006),并通过采集 100枚孔鳐卵壳(参见图1-2)进行测定的平均值相比对完全相符,参考标准参见表1。The morphological characteristics of the ray shell include total length, extreme width, body length, front horn length, rear horn length, front skirt length, rear skirt length, keel edge width, and attachment filament distance. According to the correlation of Treloar et al. The results of the study obtained the reference standard (Treloar et al., 2006) for each feature of the ray shell, and the average values measured by collecting 100 ray shells (see Figure 1-2) are completely consistent. The reference standard can be found in Table 1.
总长度(TL):卵壳前角顶端与后角顶端之间的垂直距离;Total length (TL): the vertical distance between the top of the anterior horn of the eggshell and the top of the rear horn;
极宽(MW):卵壳左侧边缘与右侧边缘之间的最大水平距离;Extreme width (MW): the maximum horizontal distance between the left and right edges of the eggshell;
体长(BL):卵壳前裙边与后裙边之间的垂直距离;Body length (BL): the vertical distance between the front skirt and the back skirt of the eggshell;
前角长(AHL):卵壳前角顶端与前裙边外缘的垂直距离;Front horn length (AHL): the vertical distance between the top of the front corner of the eggshell and the outer edge of the front skirt;
后角长(PHL):卵壳后角顶端与后裙边外缘的垂直距离;Back horn length (PHL): the vertical distance between the top of the back horn of the eggshell and the outer edge of the back skirt;
前裙边长(AAL):卵壳前裙边的外缘与内缘之间的垂直距离;Front skirt length (AAL): the vertical distance between the outer edge and the inner edge of the eggshell front skirt;
后裙边长(PAL):卵壳后裙边的外缘与内缘之间的垂直距离;Posterior skirt length (PAL): the vertical distance between the outer edge and the inner edge of the rear skirt of the eggshell;
龙骨缘宽度(LKW):卵壳龙骨缘的外缘与内缘的水平距离;Keel width (LKW): the horizontal distance between the outer and inner edges of the eggshell keel;
附着丝距离(AFL):卵壳附着丝第一附着点和第三附着点之间的垂直距离。Attachment filament distance (AFL): the vertical distance between the first attachment point and the third attachment point of the eggshell attachment filament.
表1Table 1
注:TL=总长;MW=极宽;BL=壳体长;AHL=前角长;PHL=后角长;AAL=前裙边长;PAL=后裙边长;LKW=龙骨缘宽;AFL=附着丝间距.Note: TL=total length; MW=extreme width; BL=shell length; AHL=front horn length; PHL=back corner length; AAL=front skirt length; PAL=back skirt length; LKW=keel edge width; AFL = distance between attachment wires.
实施例2Example 2
野外资源调查过程中孔鳐卵壳的快速鉴定:Rapid identification of the egg shells of the ray ray during field resource investigation:
1.选取待鉴定的软骨鱼卵壳样品13枚(依次编号1-13),需要卵壳壳体各部分完整,无污损,无缺失。附着丝基部附着点清晰可辨(图3)。1. Select 13 samples of cartilaginous fish eggshells to be identified (numbered 1-13 in sequence), and all parts of the eggshell shells must be complete without contamination or loss. The attachment point at the base of the attachment filament is clearly identifiable (Figure 3).
2.选择待鉴定的卵壳进行形态学观察初筛得到鳐鱼类卵壳:2. Select the egg shells to be identified for morphological observation and primary screening to obtain the ray fish shells:
待鉴定卵壳中多呈左右对称中央隆起的袋状结构,壳体两侧加厚形成龙骨缘,龙骨缘沿壳体长轴分别向上和向下延伸形成四个角,四个角中末段沿卵壳壳体的隆起方向延伸,每个角的末端各有一呼吸裂。同时,同一侧的两个角之间通过裙边相连(图2),为鳐类卵壳,即样品中1、2、3、5号。Most of the eggshells to be identified are pouch-shaped structures with left-right symmetrical central uplifts. The two sides of the shell are thickened to form a keel edge, which extends upward and downward along the long axis of the shell to form four corners. Extending along the uplift direction of the egg shell, each corner ends with a respiratory slit. At the same time, the two horns on the same side are connected by a skirt (Fig. 2), which is the egg shell of rays, that is, Nos. 1, 2, 3, and 5 in the sample.
同时,样品中7、9、11、12号,其卵壳的角多出现退化,同时6号的卵壳的角呈现出左右不对称结构,4号卵壳形成螺旋状,这些样品初步判定为鲨类卵壳。At the same time, among the samples No. 7, 9, 11, and 12, the corners of the eggshells were mostly degenerated, while the corners of the eggshells of No. 6 showed a left-right asymmetric structure, and the eggshells of No. 4 formed a spiral shape. These samples were preliminarily determined to be Shark shells.
另外,样品中8号卵壳呈纺锤形,壳体两侧的龙骨缘发达,并沿壳体的长轴方向向两侧延伸形成侧板,即为银鲛类卵壳。In addition, the No. 8 egg shell in the sample is spindle-shaped, and the keels on both sides of the shell are developed, and extend to both sides along the long axis of the shell to form side plates, which is the chimaera egg shell.
3.进一步将上述形态学观察初筛的鳐鱼类卵壳静置于水平桌面上,使位于卵壳四个角顶端的呼吸裂向上(图3),避免由于卵壳中水分的过度流失引起胚胎窒息死亡。3. Further place the ray fish egg shells that were initially screened by the above-mentioned morphological observation on a horizontal table, so that the respiratory cracks located at the top of the four corners of the egg shells are upward (Figure 3), to avoid excessive loss of water in the egg shells. Embryos suffocate to death.
4.辨别鳐鱼类卵壳各特征部位:分辨卵壳的前后端,以卵壳的前裙边所在的部位定义为卵壳的前端(图3),后裙边所在的部位定义为卵壳的末端。 (孔鳐卵壳的后裙边是发育成熟的仔稚幼鱼钻出的出口,发育后期的受精卵或已经孵出幼鱼的空卵壳,后裙边两薄片间常见一裂缝,而前裙边连接紧密,无裂缝。此外,与前裙边相比,后裙边的外侧边缘呈直线形,同时宽度也较宽)。4. Distinguish the characteristic parts of the ray fish egg shell: distinguish the front and rear ends of the egg shell, define the front end of the egg shell where the front skirt of the egg shell is located (Figure 3), and define the egg shell where the rear skirt is located the end. (The rear skirt of the ray shell is the exit drilled by the mature juvenile fish. For the fertilized eggs in the later stage of development or the empty egg shells that have hatched, there is often a crack between the two thin sheets of the rear skirt, while the front The skirt is tightly joined without splits. In addition, the outer edge of the rear skirt is straight and wider than the front skirt).
5.测量各初筛的鳐鱼类卵壳特征部位,并与参考标准进行对比,若各特征部位的尺寸与参考标准吻合(参见表2),即判定为孔鳐卵壳,各特征部位测量标准如下:5. Measure the characteristic parts of the ray shells of each primary screen, and compare them with the reference standard. If the size of each characteristic part is consistent with the reference standard (see Table 2), it is judged to be the egg shell of the ray, and each characteristic part is measured The standards are as follows:
总长度(TL):卵壳前角顶端与后角顶端之间的垂直距离;Total length (TL): the vertical distance between the top of the anterior horn of the eggshell and the top of the rear horn;
极宽(MW):卵壳左侧边缘与右侧边缘之间的最大水平距离;Extreme width (MW): the maximum horizontal distance between the left and right edges of the eggshell;
体长(BL):卵壳前裙边与后裙边之间的垂直距离;Body length (BL): the vertical distance between the front skirt and the back skirt of the eggshell;
前角长(AHL):卵壳前角顶端与前裙边外缘的垂直距离;Front horn length (AHL): the vertical distance between the top of the front corner of the eggshell and the outer edge of the front skirt;
后角长(PHL):卵壳后角顶端与后裙边外缘的垂直距离;Back horn length (PHL): the vertical distance between the top of the back horn of the eggshell and the outer edge of the back skirt;
前裙边长(AAL):卵壳前裙边的外缘与内缘之间的垂直距离;Front skirt length (AAL): the vertical distance between the outer edge and the inner edge of the eggshell front skirt;
后裙边长(PAL):卵壳后裙边的外缘与内缘之间的垂直距离;Posterior skirt length (PAL): the vertical distance between the outer edge and the inner edge of the rear skirt of the eggshell;
龙骨缘宽度(LKW):卵壳龙骨缘的外缘与内缘的水平距离;Keel width (LKW): the horizontal distance between the outer and inner edges of the eggshell keel;
附着丝距离(AFL):卵壳附着丝第一附着点和第三附着点之间的垂直距离。Attachment filament distance (AFL): the vertical distance between the first attachment point and the third attachment point of the eggshell attachment filament.
表2待鉴定样品测量指标Table 2 Measurement indicators of samples to be identified
通过与表1进行对比,3号样品TL、BL、PAL、AFL指标与参考标准不符;1、2、5样品各项指标与参考标准吻合,鉴定为孔鳐卵壳。By comparison with Table 1, the TL, BL, PAL, and AFL indicators of sample No. 3 did not match the reference standard; the indicators of samples 1, 2, and 5 were consistent with the reference standard, and they were identified as ray shells.
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