饲料中添加壳寡糖对斑马鱼肝肠健康和抗病能力的影响

doi:10.13304/j.nykjdb.2025.0159

中国农业科技导报 ›› 2025, Vol. 27 ›› Issue (5): 222-230.DOI: 10.13304/j.nykjdb.2025.0159

饲料中添加壳寡糖对斑马鱼肝肠健康和抗病能力的影响

符广才¹(), 李道君², 杨宏伟³, 李生辉³, 张青双³(), 张震³()

^1.广东东腾饲料有限公司，广东湛江 524094
^2.浙江湖州新路联合生物科技有限公司，浙江湖州 313300
^3.中国农业科学院饲料研究所，农业农村部饲料生物技术重点开放实验室，北京 100081

收稿日期:2025-03-06 接受日期:2025-03-18 出版日期:2025-05-15 发布日期:2025-05-20
通讯作者: 张青双,张震
作者简介:符广才E-mail：fgc.dt@163.com
基金资助:
国家自然科学基金项目(32330110)

Effects of Chitosan Oligosaccharides in Dietary on Hepatointestinal Health and Disease Resistance of Zebrafish

Guangcai FU¹(), Daojun LI², Hongwei YANG³, Shenghui LI³, Qingshuang ZHANG³(), Zhen ZHANG³()

^1.Guangdong Dongteng Feed Co. ，Ltd. ，Guangdong Zhanjiang，524094，China
^2.Zhejiang Huzhou Sino-Norway Joint Biotechnology Co. ，Ltd. ，Zhejiang Huzhou，313300，China
^3.Key Laboratory for Feed Biotechnology of Ministry of Agriculture，and Rural Affairs Institute of Feed Research，Chinese Academy of Agricultural Sciences，Beijing 100081，China

Received:2025-03-06 Accepted:2025-03-18 Online:2025-05-15 Published:2025-05-20
Contact: Qingshuang ZHANG,Zhen ZHANG

摘要/Abstract

摘要：

为探究饲料中添加壳寡糖对斑马鱼肝肠健康和抗病力的影响，设置添加质量分数0（对照）、0.05%、0.10%和0.20%壳寡糖4个处理，开展了28 d的养殖实验。结果表明，饲喂壳寡糖对斑马鱼的生长影响不显著（P>0.05）。但是，饲喂壳寡糖提高了斑马鱼的抗氧化能力，显著降低了肝脏和肠道中丙二醛的含量（P<0.05）；同时提高了肠道中抗炎因子基因（转化生长因子tgf-β）的表达，并且显著降低了促炎因子基因（核转录因子nf-κb、肿瘤坏死因子tnf-α和白介素il-1β）的表达。此外，饲喂壳寡糖提高了斑马鱼肠道中铁调素（hepcidin）、溶菌酶（LYS）、闭锁小带蛋白-1（zo-1）和闭合蛋白（occludin）基因的表达，以及在病原菌感染后的存活率。上述结果表明，饲料中添加0.10%的壳寡糖提高了斑马鱼的非特异性免疫，改善了肝、肠健康以及抗维氏气单胞菌感染的能力，为壳寡糖安全、高效地在水产养殖中的推广与利用提供了参考。

关键词: 壳寡糖, 斑马鱼, 肝肠健康, 非特异性免疫, 抗病力

Abstract:

To investigate the effects of dietary supplementation with chitosan oligosaccharide on the hepatointestinal health and disease resistance of zebrafish， a 28-day feeding trial was conducted， which designed 4 treatment groups with chitosan oligosaccharide supplementation at mass fractions of 0 （control）， 0.05%， 0.10% and 0.20%. The results showed that feeding chitosan oligosaccharide had no significant effect on the growth of zebrafish （P>0.05）. However， it enhanced the antioxidant capacity of zebrafish， significantly reduced the content of malondialdehyde in the liver and intestine （P<0.05）， increased the expression of the anti-inflammatory factor gene （transforming growth factor tgf-β） in the intestine， and significantly decreased the expression of pro-inflammatory factor genes （nuclear transcription factor nf-?κb， tumor necrosis factor tnf-?α and interleukin il-1β）. In addition， dietary chitosan oligosaccharide increased the expression of genes related to hepcidin， lysozyme （LYS）， zonula occludens-1 （zo-1）， and occladin （occludin） in the intestine of zebrafish， as well as the survival rate after pathogenic bacterial infection. The results indicated that the addition of 0.10% chitosan oligosaccharide to the feed enhances the non-specific immunity of zebrafish， improves liver and intestinal health， and the ability to resist Aeromonas veronii infection， providing a reference for the safe and efficient promotion and utilization of chitosan oligosaccharide in aquaculture.

Key words: chitosan oligosaccharides, zebrafish, liver and intestinal health, non-specific immunity, disease resistance

中图分类号:

S963

符广才, 李道君, 杨宏伟, 李生辉, 张青双, 张震. 饲料中添加壳寡糖对斑马鱼肝肠健康和抗病能力的影响[J]. 中国农业科技导报, 2025, 27(5): 222-230.

Guangcai FU, Daojun LI, Hongwei YANG, Shenghui LI, Qingshuang ZHANG, Zhen ZHANG. Effects of Chitosan Oligosaccharides in Dietary on Hepatointestinal Health and Disease Resistance of Zebrafish[J]. Journal of Agricultural Science and Technology, 2025, 27(5): 222-230.

图/表 9

表1 饲料配方与常规成分

Table 1 Feed formulation and conventional ingredient

配方/成分 Formulation/ingredient		COS₀	COS_0.05	COS_0.10	COS_0.20
配方Formulation/（g·kg^-1）	酪蛋白Casein	400.00	400.00	400.00	400.00
	明胶Gelatin	100.00	100.00	100.00	100.00
	糊精Dextrin	280.00	280.00	280.00	280.00
	猪油Lard oil	30.00	30.00	30.00	30.00
	豆油Bean oil	30.00	30.00	30.00	30.00
	赖氨酸Lysine	3.30	3.30	3.30	3.30
	VC磷酸酯VC lecithin	1.00	1.00	1.00	1.00
	多维Vitamin premix¹	4.00	4.00	4.00	4.00
	多矿Mineral premix²	4.00	4.00	4.00	4.00
	磷酸二氢钙Monocalcium phosphate	20.00	20.00	20.00	20.00
	氯化胆碱Mholine chloride	2.00	2.00	2.00	2.00
	海藻酸钠Sodium algmate	20.00	20.00	20.00	20.00
	微晶纤维素Microerystalline cellulos	40.00	40.00	40.00	40.00
	沸石粉Zeolite powder	65.70	65.20	64.70	63.70
	壳寡糖Chitosan oligosaccharide	0.00	0.50	1.00	2.00
成分Ingredient/%	粗水分Moisture	2.19	2.63	2.45	2.79
	粗灰分Crude ash	8.92	8.94	8.97	9.07
	粗脂肪Crude fat	5.85	6.18	6.10	6.10
	粗蛋白Crude protein	46.32	46.65	47.16	47.16

表2 qPCR所用引物

Table 2 Primers used in qPCR

引物名称Primer name	引物序列Primer sequence （5’-3’）	基因登录号GenBank accession No.
nfkb-F	CCGTCGTCTCTCAGCCGAT	NM_001001839
nfkb-R	AGAAGCGCACCTCAATGTCC	NM_001001839
il-1β-F	TGGACTTCGCAGCACAAAATG	NM_212844
il-1β-R	GTTCACTTCACGCTCTTGGATG	NM_212844
tnf-α-F	GCGCTTTTCTGAATCCTACG	NM_212859
tnf-α-R	TGCCCAGTCTGTCTCCTTCT	NM_212859
tgf-β-F	CAAACCAGCAGAGCACGGATA	NM_182873
tgf-β-R	CGGTGCCAACAGCTCGTCTC	NM_182873
il-10-F	TCACGTCATGAACGAGATCC	NM_001020785
il-10-R	CCTCTTGCATTTCACCATATCC	NM_001020785
rps11-F	ACAGAAATGCCCCTTCACTG	NM_213377
rps11-R	GCCTCTTCTCAAAACGGTTG	NM_213377
hepcidin-F	GCAGGAGAACCAACATCTGAC	NM_001289794
hepcidin-R	CAGAATTTGCAGCAGTATCCG	NM_001289794
LYS-F	GATTTGAGGGATTCTCCATTGG	NM_139180
LYS-R	CCGTAGTCCTTCCCCGTATCA	NM_139180
C3b-F	CGTCTCCGTACACCATCCATT	NM_131243
C3b-R	GGCGTCTCATCAGGATTTGTTAC	NM_131243

图1 不同处理斑马鱼的生长性能

Fig. 1 Growth performance of zebrafish under different treatments

图2 不同处理斑马鱼肠道炎症因子相关基因的表达注：图中不同小写字母表示P<0.05差异显著。

Fig. 2 Expression of genes related to intestinal inflammatory factors in zebrafish under different treatmentsNote： Different lowercase letters in the figure indicate significant difference at P<0.05 level.

图3 不同处理斑马鱼肠道紧密连接相关基因的表达注：图中不同小写字母表示P <0.05差异显著。

Fig. 3 Expression of genes related to intestinal tight junctions in zebrafish under different treatmentsNote： Different lowercase letters in the figure indicate significant difference at P<0.05 level.

图4 不同处理斑马鱼肠道抗菌相关基因的表达注：图中不同小写字母表示P<0.05差异显著。

Fig. 4 Expression of genes related to intestinal antimicrobial activity in zebrafish under different treatments.Note：Different lowercase letters in the figure indicate significant difference at P<0.05 level.

图5 不同处理斑马鱼肝脏的抗氧化水平注：图中不同小写字母表示P <0.05差异显著。

Fig. 5 Antioxidant levels in the liver of zebrafish under different treatmentsNote： Different lowercase letters in the figure indicate significant difference at P<0.05 level.

图6 不同处理斑马鱼肠道的抗氧化水平注：图中不同小写字母表示P <0.05差异显著。

Fig. 6 Antioxidant levels in the intestines of zebrafish under different treatmentsNote： Different lowercase letters in the figure indicate significant difference at P<0.05 level.

图7 不同处理斑马鱼抗细菌攻击的能力

Fig. 7 Ability of zebrafish to resist bacterial challenge under different treatments

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饲料中添加壳寡糖对斑马鱼肝肠健康和抗病能力的影响

Effects of Chitosan Oligosaccharides in Dietary on Hepatointestinal Health and Disease Resistance of Zebrafish

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