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<p>Distiller’s dried grains with solubles (DDGS) were subjected to compound enzymatic deep hydrolysis (CEDEH) and evaluated as a plant-protein substitute for fish meal in zebrafish diets. The CEDEH protocol sequentially applied cellulase and α-amylase followed by proteolysis using neutral and flavor proteases at a 2:1 ratio. Enzyme-treated DDGS were incorporated into experimental diets to partially replace fish meal at 0%, 10%, and 20% inclusion levels. Six dietary treatments were established: DNP10 and DNP20 (neutral protease–hydrolyzed DDGS at 10% and 20%), DFP10 and DFP20 (flavor protease–hydrolyzed DDGS at 10% and 20%), and DCP10 and DCP20 (mixed protease–hydrolyzed DDGS at 10% and 20%). A 30-day feeding trial was conducted on 2-month-old zebrafish to assess physiological responses, intestinal histology, digestive enzyme activity, and gut microbiome–metabolome responses. Notably, CEDEH processing significantly enhanced duodenal villus height (<i>P</i> &lt; 0.01), indicating improved nutrient absorption capacity. At 10% substitution (DCP10 group), hepatic antioxidant capacity significantly improved, with superoxide dismutase (SOD) and glutathione (GSH) activities increasing by 24.8% and 90.8%, respectively, compared to controls (<i>P</i> &lt; 0.05). Gut microbiota restructuring demonstrated functional probiotics enrichment, including <i>Cellulomonas</i> (+ 57.3%) and anti-inflammatory <i>Tabrizicola</i> (+ 354.33%). Concurrently, key digestive enzymes (amylase and trypsin) activities in the duodenum markedly increased by 32.1% and 27.4%. Metagenomic analysis revealed upregulation of key metabolic pathways (ko00020 TCA cycle; ko00290 branched-chain amino acid synthesis). Importantly, 20% substitution (DCP20) preserved intestinal integrity (goblet cells increased by &gt; 40%) without compromising growth performance. These results confirm that CEDEH-treated DDGS is a suitable fish meal substitute at inclusion levels ≤ 20%, promoting host metabolism and disease resistance.</p>

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Intestinal health and metabolic balance of zebrafish (Danio rerio) fed on diets containing enzymatically hydrolyzed distiller’s dried grains with solubles (DDGS)

  • 期刊论文

  • 发表日期：

    2025年10月15日

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  • Chun Wang,
  • Xin Peng,
  • Ling Ao,
  • Shan Zeng,
  • Jianping Yang,
  • Caihong Shen,
  • Mengyan Wang,
  • Xiaolong Yao,
  • Yingxue Sun

摘要

Distiller’s dried grains with solubles (DDGS) were subjected to compound enzymatic deep hydrolysis (CEDEH) and evaluated as a plant-protein substitute for fish meal in zebrafish diets. The CEDEH protocol sequentially applied cellulase and α-amylase followed by proteolysis using neutral and flavor proteases at a 2:1 ratio. Enzyme-treated DDGS were incorporated into experimental diets to partially replace fish meal at 0%, 10%, and 20% inclusion levels. Six dietary treatments were established: DNP10 and DNP20 (neutral protease–hydrolyzed DDGS at 10% and 20%), DFP10 and DFP20 (flavor protease–hydrolyzed DDGS at 10% and 20%), and DCP10 and DCP20 (mixed protease–hydrolyzed DDGS at 10% and 20%). A 30-day feeding trial was conducted on 2-month-old zebrafish to assess physiological responses, intestinal histology, digestive enzyme activity, and gut microbiome–metabolome responses. Notably, CEDEH processing significantly enhanced duodenal villus height (P < 0.01), indicating improved nutrient absorption capacity. At 10% substitution (DCP10 group), hepatic antioxidant capacity significantly improved, with superoxide dismutase (SOD) and glutathione (GSH) activities increasing by 24.8% and 90.8%, respectively, compared to controls (P < 0.05). Gut microbiota restructuring demonstrated functional probiotics enrichment, including Cellulomonas (+ 57.3%) and anti-inflammatory Tabrizicola (+ 354.33%). Concurrently, key digestive enzymes (amylase and trypsin) activities in the duodenum markedly increased by 32.1% and 27.4%. Metagenomic analysis revealed upregulation of key metabolic pathways (ko00020 TCA cycle; ko00290 branched-chain amino acid synthesis). Importantly, 20% substitution (DCP20) preserved intestinal integrity (goblet cells increased by > 40%) without compromising growth performance. These results confirm that CEDEH-treated DDGS is a suitable fish meal substitute at inclusion levels ≤ 20%, promoting host metabolism and disease resistance.