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

doi:10.13304/j.nykjdb.2025.0159

Abstract

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

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

CLC Number:

S963

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.

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.

Figures/Tables 9

References 23

1	BALDWIN L. The effects of stocking density on fish welfare [J]. Plymouth Student Scientist, 2011, 4:372-383.
2	RINGØ E, DIMITROGLOU A, HOSEINIFAR S H, et al.. Prebiotics in finfish: an update [J/OL]. Aquac. Nutr., 2014,360-400 [2025-03-14]. .
3	WANG T, WU H X, LI W J, et al.. Effects of dietary mannan oligosaccharides (MOS) supplementation on metabolism, inflammatory response and gut microbiota of juvenile Nile Tilapia (Oreochromis niloticus) fed with high carbohydrate diet [J]. Fish Shellfish Immunol., 2022, 130: 550-559.
4	张廷军.甲壳素/壳聚糖及其衍生物的应用情况[J].水产科技情报,1999(6):243-247.
5	王爱勤,杨立明.天然高分子材料壳聚糖的研究应用进展[J].化工新型材料,1994,22(9):9-12, 24.
6	LODHI G, KIM Y S, HWANG J W, et al.. Chitooligosaccharide and its derivatives: preparation and biological applications [J/OL]. Biomed. Res. Int., 2014, 654913 [2025-03-14]. .
7	赵雅.壳寡糖对断奶仔猪机体免疫力的影响[J].国外畜牧学(猪与禽),2024,44(4):62-64.
8	刘叶青.枯草芽孢杆菌和壳寡糖对肉鸡抗氧化、免疫和肠道屏障功能的影响[D].内蒙古:内蒙古农业大学,2023.
	LIU Y Q. Effects of Bacillus subtilis and chito-oligosaccharides on antioxidant, immune and intestinal barrier function in broilers [D]. Hohhot: Inner Mongolia Agricultural University.
9	辛国省.壳寡糖在宠物犬食品中的应用开发[Z].2023-09-05.
10	TEAME T, WU X X, HAO Q, et al.. Dietary SWF^® enhanced growth performance and disease resistance in hybrid sturgeon (Acipenser baerii x Acipenser schrenckii) mediated by the gut microbiota [J/OL]. Aquac. Rep., 2020, 17: 100346 [2025-03-14]. .
11	LI S, YANG H, JIN Y, et al.. Dietary cultured supernatant mixture of Cetobacterium somerae and Lactococcus lactis improved liver and gut health, and gut microbiota homeostasis of zebrafish fed with high-fat diet [J/OL]. Fish Shellfish Immunol., 2023, 142: 109139 [2025-03-14]. .
12	王艳丽,李琦,张文静,等.饲料中添加不同浓度的壳寡糖对黄河鲤幼鱼生长及抗氧化功能的影响[J].水产学杂志,2024,37(3):58-65.
	WANG Y L, LI Q, ZHANG W J, et al.. Effects of different levels of dietary chitooligosaccharides on growth and antioxidant function of juvenile Yellow River common carp [J]. Chin. J. Fish., 2024, 37(3): 58-65.
13	胡晓伟,上官静波,黎中宝,等.饲料中添加壳寡糖对花鲈(Lateolabrax japonicus)幼鱼的生长、消化和血清生化指标的影响[J].海洋学报,2018,40(2):69-76.
	HU X W, SHANGGUAN J B, LI Z B, et al.. Effects of dietary Chitosan oligosaccharide on the performance, digestion and serum biochemical indexes of the juvenile Japanese seabass(Lateolabrax japonicus) [J]. Haiyang Xuebao,2018,40(2):69-76.
14	田娟,孙立威,文华,等.壳寡糖对吉富罗非鱼幼鱼生长性能、前肠组织结构及肠道主要菌群的影响[J].中国水产科学,2013,20(3):561-568.
	TIAN J, SUN L W, WEN H, et al.. Effect of dietary chitooligosaccharides on growth performance, gut flora and intestinal structure of juvenile GIFT Tilapia oreochromis niloticus [J]. J. Fish. Sci. China, 2013, 20(3): 561-568.
15	李振达.壳寡糖的制备及其在水产养殖上的应用研究[D]. 浙江:浙江海洋学院,2011.
	LI Z D. The preparation of chitooligosaccharides and the application in aquaculture [D]. Zhejiang: Zhejiang Ocean University,2011.
16	常孟阳,杨越,李晨露,等.虾青素对镉离子(Cd²⁺)诱导下斑马鱼胚胎抗氧化指标的影响[J].水产学杂志,2024,37(3):37-43, 49.
	CHANG M Y, YANG Y, LI C L, et al.. Effects of astaxanthin on antioxidant parameters of zebrafish embryos induced by Cd²⁺ [J]. Chin. J. Fish.,2024,37(3):37-43, 49.
17	马兴右,朱志,罗雪莲,等.黄精多糖对维氏气单胞菌抑制效果研究[J].水产养殖,2024,45(6):30-36.
	MA X Y, ZHU Z, LUO X L, et al.. Study on the inhibitory effect of Polygonatum sibiricum polysaccharides on Aeromonas veronii [J]. J. Aquac., 2024,45(6):30-36.
18	郭印,戴习林.急性盐度胁迫对美洲鲥幼鱼渗透调节的影响[J].水产科学,2022,41(4):676-681.
	GUO Y, DAI X L. Effects of acute salinity stress on osmoregulation of juvenile American shad Alosa sapidissima [J]. Fish. Sci., 2022, 41(4): 676-681.
19	施斐,黄垚,卢志杰,等.壳寡糖对虎龙斑的生长、免疫及肠道菌群的影响[J].水产学报,2022,46(9):1689-1700.
	SHI F, HUANG Y, LU Z J, et al.. Effects of oligochitosan on the growth, immune responses and gut microbes of hybrid groupers [Epinephelus fuscoguttatus (♀) × E. lanceolatu (♂)] [J]. J. Fish. China, 2022, 46(9): 1689-1700.
20	苏鹏.壳寡糖对红鳍东方鲀(Takifugu rubripes)生长、免疫酶及肠道菌群的影响[D].大连:大连海洋大学,2016.
	SU P. The effects of Chitosan-oligosaccbarides on the growth performances, non-pecitify enzymes and intestinal bacteria flora of Takifugu rubripes [D]. Dalian: Dalian Ocean University.2016.
21	张婷婷.壳寡糖对大口黑鲈生长、肝肠健康的影响及肝脏转录组学分析[D].重庆:西南大学,2024.
	ZHANG T T. Effects of chito-oilgosaccharides on growth,hepatointestinal health and liver transcriptomic analysis of largemouth bass Micropterus salmoides [D].Chongqing: Southwest University,2024.
22	LIN S, MAO S, GUAN Y, et al.. Dietary administration of chitooligosaccharides to enhance growth, innate immune response and disease resistance of Trachinotus ovatus [J]. Fish Shellfish Immunol., 2012, 32(5): 909-913.
23	GUAN D, SUN H, MENG X, et al.. Effects of different molar mass chitooligosaccharides on growth, antioxidant capacity, non-specific immune response, and resistance to Aeromonas hydrophila in GIFT Tilapia oreochromis niloticus [J]. Fish Shellfish Immunol., 2019, 93: 500-507.

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

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

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

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

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[5]	ZOU Yuan-yuan, LIU Yang, ZHAO Liang, LIU Lin, SONG Wei*. Research on the Diversity of Indigenous Bacterial Community in #br# the Seeds of Two Rice Varieties with Different Disease-resistance [J]. Journal of Agricultural Science and Technology, 2016, 18(4): 9-16.
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[7]	WANG Fajun1,2,3, GAO Ying1,2*, ZHAO Kaijun1,2. Research Progress on Plant Signaling Molecules Involved in Disease Resistance [J]. , 2013, 15(5): 87-92.
[8]	LI Dong-xiao1,2, ZHANG Guo-guang2, GUO Li-jia2, LI Min2, MAO Qian2, WANG Wei2, . Studies on Rice Resistance-related Proteins in Response to Bacterial Leaf Streak, Xanthomonas Oryzae pv. Oryzicola [J]. , 2010, 12(5): 62-67.
[9]	LI Zao-xia\|ZHAO Jun. Regulation of Nitric Oxide in Plant Drought Tolerance and Disease Resistance [J]. , 2008, 10(S1): 7-11.
[10]	YU Hai-qin, JIAO Fang-chan, XIAO Bing-guang, Li De-tuan, LU Xiu-ping . Studies on Identification of Resistance to Black Shank Disease of \|Tobacco Germplasm Resources in Seedling Stage [J]. , 2008, 10(4): 70-75.
[11]	PEI Xin-wu, JIA Shi-rong . Progress in Genetic Engineering of Banana for \|Fungal Disease Resistance [J]. , 2006, 8(4): 1-7.