一种显著提高实验用斑马鱼繁殖效率和使用寿命的优化养殖方案

doi:10.12300/j.issn.1674-5817.2023.004

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (3): 297-306.DOI: 10.12300/j.issn.1674-5817.2023.004

一种显著提高实验用斑马鱼繁殖效率和使用寿命的优化养殖方案

金仕容¹(), 华叶¹(), 訾化星², 杜旭飞¹, 卜纪雯²()()

^1.中国科学院脑科学与智能技术卓越创新中心(原中国科学院上海生命科学院神经科学研究所), 全脑介观神经联接图谱研究平台(斑马鱼), 上海 200031
^2.中国科学院脑科学与智能技术卓越创新中心(原中国科学院上海生命科学院神经科学研究所), 感觉整合与行为研究组, 上海 200031

收稿日期:2023-01-16 修回日期:2023-04-18 出版日期:2023-06-25 发布日期:2023-07-18
通讯作者: 卜纪雯（1982—），女，硕士，高级实验师，研究方向：分子发育生物学。E-mail：jwbu@ion.ac.cn。ORCID：0009-0006-0933-7314
作者简介:金仕容（1994—），女，硕士，实验师，研究方向：水产养殖。E-mail：18817772747@163.com；
华叶（1983—），女，硕士，助理研究员，研究方向：斑马鱼早胚发育与精子发生。E-mail：ye.hua@ion.ac.cn
基金资助:
国家重点研发计划“发育编程及其代谢调节”重点专项项目“斑马鱼发育与代谢突变体库的系统创制”(2018YFA0801000)

An Optimized Experimental Zebrafish Breeding Scheme for Significantly Enhancing Reproductive Efficiency and Service Life

Shirong JIN¹(), Ye HUA¹(), Huaxing ZI², Xufei DU¹, Jiwen BU²()()

^1.Center for Excellence in Brain Science and Intelligence Technology (Institute of Neuroscience), Chinese Academy of Sciences, Research Platform of Whole Brain Mesoscopic Neural Junction Map (Zebrafish), Shanghai 200031, China
^2.Center for Excellence in Brain Science and Intelligence Technology (Institute of Neuroscience), Chinese Academy of Sciences, Lab of Sensory Integration & Behavior, Shanghai 200031, China

Received:2023-01-16 Revised:2023-04-18 Published:2023-06-25 Online:2023-07-18
Contact: BU Jiwen (ORCID: 0009-0006-0933-7314), E-mail: jwbu@ion.ac.cn

摘要/Abstract

摘要：

目的解决实验用斑马鱼出现的生长发育滞后、产卵量不足等问题，以提高实验用斑马鱼的繁殖效率和使用寿命。方法取受精后2月龄的斑马鱼，以投喂商品化的观赏鱼专用颗粒饲料或冰冻成体卤虫为实验组，以投喂活的幼体卤虫为对照组。不同饵料投喂70 d内，通过测量体长和体重评估斑马鱼生长性能，通过统计产卵量和产卵率等评估斑马鱼繁殖性能。对甲状腺肿大病症明显的斑马鱼投喂颗粒饲料，通过测量其甲状腺肿大病灶直径大小变化评估颗粒饲料对该病症的抑制效果。将3种饲料配合使用，对饲养方案进行优化，通过繁殖性能测试验证其用于斑马鱼饲养的实际效果。结果从60 dpf开始投喂颗粒饲料直到111 dpf，颗粒饲料组的体长、体重均日渐优于对照组（P<0.000 1）。从60 dpf开始投喂成体卤虫直到96 dpf，成体卤虫组的体长增长趋势与对照组基本相同，而75～82 dpf时成体卤虫组的雌鱼体重极明显高于对照组的雌鱼（P<0.000 1）。成体卤虫组斑马鱼相比于对照组，雌雄间体色差异显著，在75 dpf时，已可以通过体色差异准确区分性别；而且成体卤虫组斑马鱼3月龄时产卵率显著高于对照组（94.44% vs 27.78%，P<0.05）。另外，投喂颗粒饲料130 d后，所有实验用斑马鱼的甲状腺肿大病灶均消失。根据以上结果，将3种饲料组合，优化大于2月龄的斑马鱼饲养方案为上午保持喂食幼体卤虫，下午用颗粒饲料和成体卤虫交替投喂，直至12月龄斑马鱼的产卵率可以保持70%，产卵量达到（233.6±3.95）颗，受精率和孵化率分别为97.47%和90.24%，均显著高于原方案饲养组（P<0.001或P=0.01）。结论相比幼体卤虫饵料，商品化的颗粒饲料能显著提升斑马鱼的生长性能，并对甲状腺肿大病症有治疗作用，而成体卤虫饵料能显著提高斑马鱼的早期繁殖性能。优化后的饲养方案成功提高了斑马鱼的产卵效率，并延长其繁殖使用寿命，能更好地保障相关科学研究的开展。

关键词: 斑马鱼, 饲养方案, 繁殖性能, 生长性能

Abstract:

Objective To solve the problems of delayed growth and development and insufficient spawning of experimental Zebrafish, so as to improve the reproductive efficiency and service life of experimental Zebrafish. Methods The zebrafish at the age of 2 months after fertilization were divided into two groups. The experimental group was fed with dry commercial diets specifically designed for ornamental fish or frozen adult brine shrimp, while control group was fed with live laval brine shrimp. Within a period of 70 days, the growth performance of the zebrafish was evaluated by measuring body length and weight, and the reproductive performance was assessed by measuring the fecundity and spawning rate. Zebrafish with apparent goiter disease were fed with dry commercial diets, and the inhibitory effect of the pellets on this disease was evaluated by measuring the diameter of the thyroid enlargement lesion. The three feeding methods were combined, and the feeding plan was optimized. The actual effects of the plan on zebrafish rearing were validated through reproductive performance tests. Results Starting from 60 days post-fertilization (dpf) until 111 dpf, the body length and weight of the dry commercial diets feed group gradually surpassed those of control group (all P<0.000 1). From 60 dpf to 96 dpf, the growth trend in body length of the adult brine shrimp group was similar to that of control group, but the female fish in the adult brine shrimp group had significantly higher body weight than the female fish in control group at 75-82 dpf (P<0.000 1). Compared to control group, there was a significant difference in body color between males and females in the adult brine shrimp group, and at 75 dpf, gender could be accurately distinguished by body color differences. Furthermore, the spawning rate of the zebrafish in the adult brine shrimp group at 3 months of age was significantly higher than that of control group (94.44% vs. 27.78%, P<0.05). Additionally, after feeding with the dry commercial diets for 130 days, all thyroid enlargement lesions in the experimental zebrafish disappeared. Based on the above results, the three feeding methods were combined and the feeding plan for zebrafish older than 2 months of age was optimized as follows: feed live brine shrimp in the morning, and alternate between dry commercial diets and adult brine shrimp in the afternoon. This feeding plan lasts until the age of 12 months. The spawning rate of Zebrafish can maintan 70%, and the spawning amount can reach (233.6±3.95) eggs. The fertilization rate and hatching rate were 97.47% and 90.24%, respectively, both significantly higher than those of control group (P<0.001, or P=0.01). Conclusion Compared to live brine shrimp feed, the dry commercial diets feed significantly improves the growth performance of zebrafish and has a therapeutic effect on thyroid enlargement disease. On the other hand, adult brine shrimp feed significantly enhances the early reproductive performance of zebrafish. The optimized feeding plan successfully improves the spawning efficiency of laboratory zebrafish, prolonging their reproductive lifespan and better supporting relevant scientific research.

Key words: Zebrafish, Breeding scheme, Reproduction, Growth

中图分类号:

Q95-33

金仕容, 华叶, 訾化星, 杜旭飞, 卜纪雯. 一种显著提高实验用斑马鱼繁殖效率和使用寿命的优化养殖方案[J]. 实验动物与比较医学, 2023, 43(3): 297-306.

Shirong JIN, Ye HUA, Huaxing ZI, Xufei DU, Jiwen BU. An Optimized Experimental Zebrafish Breeding Scheme for Significantly Enhancing Reproductive Efficiency and Service Life[J]. Laboratory Animal and Comparative Medicine, 2023, 43(3): 297-306.

图/表 5

表1 不同饵料营养成分表

Table 1 Nutrition ingredients of different diets

营养成分 Nutrition ingredients	需求量 Requirement	幼体卤虫(活) Larva brine shrimp (live)	成体卤虫(冻) Adult brme shrimp (frozen)	颗粒饲料1 Dry commercial diets-1	颗粒饲料2 Dry commercial diets-2
钙Calcium/%	0.9～1.6 (斑马鱼)^[5]	未检出	未检出	2.31	2.62
磷Phosphorus/%	0.8～1.4 (斑马鱼)^[5]	0.12	0.07	1.92	2.22
镁Magnesium/(mg·kg^-1)	600 mg/kg (鲤鱼)^[14]	186	102	6 110	5 360
铁Iron/(mg·kg^-1)	150 mg/kg (鲤鱼)^[15]	未检出	58.5	575	940
硒Selenium/(mg·kg^-1)	0.12 mg/kg (鲤鱼)^[16]	0.15	0.27	3.22	4.55
锌Zinc/(mg·kg^-1)	15～30 mg/kg (鲤鱼)^[15]	20	5.3	160	300
维生素B3 Vitamin B3/(mg·kg^-1)	30～40 mg/kg (鲤鱼)^[17]	未检出	133	未检出	未检出
维生素C Vitamin C/(mg·kg^-1)	30～50 mg/kg (鲤鱼)^[17]	50.5	102	1 000	3 960
维生素B2 Vitamin B2/(mg·kg^-1)	7～10 mg/kg (鲤鱼)^[17]	未检出	未检出	38	84.8

图1 颗粒饲料和成体卤虫对斑马鱼生长的影响注：每组9条斑马鱼，数据均以平均值±标准差表示。A～B为颗粒饲料组和幼体卤虫组斑马鱼的体长（A）和体重（B）变化（与幼体卤虫组相比，**P<0.01，*P<0.05）。C～D为成体卤虫组和幼体卤虫组雌雄斑马鱼的体长（C）和体重（D）变化（分别与幼体卤虫组雌雄斑马鱼相比，****P<0.000 1，***P<0.001，**P<0.01，*P<0.05）。dpf，受精后日龄。

Figure 1 Effects of dry commercial diets and adult brine shrimp on growth of zebrafishNote： Data are presented as means±SEM （n=9）. A - B are the body length （A） and body weight （B） changes of zebrafish in dry commercial diets group and larva brine shrimp group （compared with larva brine shrimp group， **P<0.01， *P<0.05）. C-D are the changes in body length （C） and body weight （D） of male and female zebrafish in adult brine shrimp group and larva brine shrimp group （compared with male and female zebrafish in larval brine shrimp group， ****P<0.000 1， ***P<0.001， **P<0.01， *P<0.05）. dpf， age after fertilization.

图2 不同饲料对斑马鱼繁殖性能的影响注：A，饲喂成体卤虫组、幼体卤虫组或颗粒饲料组斑马鱼的典型外观（♂雄，♀雌）。B，饲喂成体卤虫或幼体卤虫组的3月龄和5月龄斑马鱼产卵率、产卵量、受精率和孵化率比较（每组9条斑马鱼，产卵量、受精率和孵化率均以平均值±标准差表示，**P<0.01］。

Figure 2 Effects of different diets on reproductive characteristics of zebrafishNote： A, the typical appearance of zebrafish (♂ male, ♀ female) in dry commercial diets group, adult brine shrimp group and larva brine shrimp group. B is the comparison of spawning rate, spawning amount, fertilization rate and hatching rate of 3-month-old and 5-month-old zebrafish between adult brine shrimp group and larva brine shrimp group （9 zebrafish in each group. Spawning amount, fertilization rate and hatching rate are presented as means±SEM, **P<0.01).

图3 颗粒饲料对斑马鱼甲状腺肿大病症的影响注：A，甲状腺肿大的斑马鱼外观；B，颗粒饲料喂养后的斑马鱼甲状腺直径变化[数据均以平均值±标准差表示（n=15），??P<0.01，????P<0.000 1，ns表示没有统计学差异]。

Figure 3 Effect of dry commercial diets on goiter disease of zebrafishNote: A, Appearance of Zebrafish with goiter; B, Changes in thyroid diameter of Zebrafish after dry commercial diets feeding [Data are presented as means ±SEM (n=15), ?P<0.01, ????P<0.000 1, ns indicated no statistical difference].

图4 新饲养方案Du-Lab Zeb's Recipe 2.0示意图

Figure 4 New breeding scheme Du-Lab Zeb's Recipe 2.0

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一种显著提高实验用斑马鱼繁殖效率和使用寿命的优化养殖方案

An Optimized Experimental Zebrafish Breeding Scheme for Significantly Enhancing Reproductive Efficiency and Service Life

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