Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)

This study aims to investigate the effects of dietary supplementation with protease on the growth performance, liver health, immunity, and intestinal microbiota of grass carp. Three levels of protease, 0 U/kg (P0), 6000 U/kg (P6000) and 12,000 U/kg (P12000), were formulated into diets using post-spr...

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Main Authors: Tingyao Zhu, Lihui Sun, Qian Li, Jianhu Jiang, Haiqi Zhang, Zhangying Ye, Jianming Chen, Jianlin Guo
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Aquaculture Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352513425002649
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author Tingyao Zhu
Lihui Sun
Qian Li
Jianhu Jiang
Haiqi Zhang
Zhangying Ye
Jianming Chen
Jianlin Guo
author_facet Tingyao Zhu
Lihui Sun
Qian Li
Jianhu Jiang
Haiqi Zhang
Zhangying Ye
Jianming Chen
Jianlin Guo
author_sort Tingyao Zhu
collection DOAJ
description This study aims to investigate the effects of dietary supplementation with protease on the growth performance, liver health, immunity, and intestinal microbiota of grass carp. Three levels of protease, 0 U/kg (P0), 6000 U/kg (P6000) and 12,000 U/kg (P12000), were formulated into diets using post-spray technology. After 9 weeks of feeding, the P6000 group had a higher weight gain rate than the P0 group, with both P6000 and P12000 groups exhibiting higher protein retention ratio and lower feed conversion ratio compared to P0. As protease levels increased, crude protein content and intestinal protease activity increased, while alkaline phosphatase (ALP) content decreased. Concurrently, the P6000 group showed higher aspartate transaminase (AST) and triglyceride (TG) levels but lower amylase content compared to P0. In terms of immune indicators, the expression levels of Intelectin and MHC-II β in the P6000 and P12000 groups were significantly lower than in P0, and IgM expression gradually increased with increasing protease levels. After challenge with Aeromonas veronii, the P12000 group showed higher survival rate. The histological results indicated that the intestinal villi length in the P6000 group was notably longer than in other groups. The intestinal microbiota analysis revealed that with increasing protease levels, Fusobacteriota abundance decreased and Actinobacteriota and Cyanobacteria abundance increased at the phylum level, while Aeromonas relative abundance decreased at the genus level. Overall, 6000 U/kg protease boosts grass carp growth and nutrient retention maximally, while 12,000 U/kg protease increases survival but may increase liver metabolic burden. Given these outcomes, the optimal protease inclusion level for grass carp aquaculture is approximately 6000 U/kg.
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spelling doaj-art-eb5ccaa3edbd4496b3ade5e63ac136a72025-08-20T03:40:50ZengElsevierAquaculture Reports2352-51342025-09-014310287810.1016/j.aqrep.2025.102878Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)Tingyao Zhu0Lihui Sun1Qian Li2Jianhu Jiang3Haiqi Zhang4Zhangying Ye5Jianming Chen6Jianlin Guo7Agriculture and Rural Affairs Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China; College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310000, ChinaAgriculture and Rural Affairs Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, ChinaAgriculture and Rural Affairs Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, ChinaAgriculture and Rural Affairs Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, ChinaAgriculture and Rural Affairs Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, ChinaCollege of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310000, ChinaAgriculture and Rural Affairs Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, ChinaAgriculture and Rural Affairs Ministry Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China; Corresponding author.This study aims to investigate the effects of dietary supplementation with protease on the growth performance, liver health, immunity, and intestinal microbiota of grass carp. Three levels of protease, 0 U/kg (P0), 6000 U/kg (P6000) and 12,000 U/kg (P12000), were formulated into diets using post-spray technology. After 9 weeks of feeding, the P6000 group had a higher weight gain rate than the P0 group, with both P6000 and P12000 groups exhibiting higher protein retention ratio and lower feed conversion ratio compared to P0. As protease levels increased, crude protein content and intestinal protease activity increased, while alkaline phosphatase (ALP) content decreased. Concurrently, the P6000 group showed higher aspartate transaminase (AST) and triglyceride (TG) levels but lower amylase content compared to P0. In terms of immune indicators, the expression levels of Intelectin and MHC-II β in the P6000 and P12000 groups were significantly lower than in P0, and IgM expression gradually increased with increasing protease levels. After challenge with Aeromonas veronii, the P12000 group showed higher survival rate. The histological results indicated that the intestinal villi length in the P6000 group was notably longer than in other groups. The intestinal microbiota analysis revealed that with increasing protease levels, Fusobacteriota abundance decreased and Actinobacteriota and Cyanobacteria abundance increased at the phylum level, while Aeromonas relative abundance decreased at the genus level. Overall, 6000 U/kg protease boosts grass carp growth and nutrient retention maximally, while 12,000 U/kg protease increases survival but may increase liver metabolic burden. Given these outcomes, the optimal protease inclusion level for grass carp aquaculture is approximately 6000 U/kg.http://www.sciencedirect.com/science/article/pii/S2352513425002649Exogenous proteaseGrass carpGrowth performanceImmune responseIntestinal microbiota
spellingShingle Tingyao Zhu
Lihui Sun
Qian Li
Jianhu Jiang
Haiqi Zhang
Zhangying Ye
Jianming Chen
Jianlin Guo
Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)
Aquaculture Reports
Exogenous protease
Grass carp
Growth performance
Immune response
Intestinal microbiota
title Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)
title_full Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)
title_fullStr Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)
title_full_unstemmed Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)
title_short Effects of exogenous protease supplementation on growth performance, liver health, immunity and intestinal microbiota of grass carp (Ctenopharyngodon idella)
title_sort effects of exogenous protease supplementation on growth performance liver health immunity and intestinal microbiota of grass carp ctenopharyngodon idella
topic Exogenous protease
Grass carp
Growth performance
Immune response
Intestinal microbiota
url http://www.sciencedirect.com/science/article/pii/S2352513425002649
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