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<p>This study optimized the freeze-drying process for the <i>Labeo rohita</i> gut-derived probiotic <i>Bacillus amyloliquefaciens</i> COFCAU_P1 and assessed its immunoprotective effects in <i>L. rohita</i>. Freeze-drying stabilizes probiotics but often reduces viability. To improve preservation, 24 protectant combinations were tested, with 10% trehalose and 2% gum Arabic proving the most effective. This combination maintained 99.02% viability immediately after freeze-drying and 92.44% and 91.87% after 180&#xa0;days at room temperature and 4&#xa0;°C, respectively. Scanning electron microscopy confirmed structural integrity. The optimized freeze-dried formulation (BAFP) exhibited superior thermal, pH, and bile tolerance compared to its fresh form (BA), retained antimicrobial activity against aquaculture pathogens, and preserved digestive enzyme production, hydrophobicity, aggregation, and antioxidant properties. Feeding <i>L. rohita</i> with BAFP (1 × 10⁹ cells g<sup>−1</sup> feed) for 28&#xa0;days significantly enhanced serum NBT, anti-protease, myeloperoxidase activities, albumin, and total protein while reducing glucose, SGOT, SGPT, CK-MB, and alkaline phosphatase. It also upregulated immune genes (<i>TNF-α, C3, Lysozyme, IFN-ɣ</i>) and downregulated <i>iNOS</i> in the head-kidney and liver. Challenge trials with <i>Aeromonas hydrophila</i> ATCC 7966, <i>A. salmonicida</i> subspecies <i>salmonicida</i> COFCAU_AS, and <i>A. veronii</i> COFCAU_AV1 showed significantly higher survival in BAFP-fed fish. These findings demonstrate that the optimized freeze-dried <i>B. amyloliquefaciens</i> formulation enhances probiotic stability and boosts host immunity, offering effective protection against bacterial pathogens in aquaculture.</p>

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Freeze-drying of Bacillus amyloliquefaciens COFCAU_P1 probiotics: process optimization, and evaluation of its immunoprotective role in Labeo rohita

  • 期刊论文

  • 发表日期：

    2025年3月24日

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  • Tanmoy Gon Choudhury,
  • Sangita Roy,
  • Suraj Kumar,
  • Debaprasad Rath,
  • Supratim Malla,
  • Partha Sarathi Tripathy,
  • Dibyendu Kamilya,
  • Janmejay Parhi,
  • Ratan Kumar Saha,
  • Arun Bhai Patel

摘要

This study optimized the freeze-drying process for the Labeo rohita gut-derived probiotic Bacillus amyloliquefaciens COFCAU_P1 and assessed its immunoprotective effects in L. rohita. Freeze-drying stabilizes probiotics but often reduces viability. To improve preservation, 24 protectant combinations were tested, with 10% trehalose and 2% gum Arabic proving the most effective. This combination maintained 99.02% viability immediately after freeze-drying and 92.44% and 91.87% after 180 days at room temperature and 4 °C, respectively. Scanning electron microscopy confirmed structural integrity. The optimized freeze-dried formulation (BAFP) exhibited superior thermal, pH, and bile tolerance compared to its fresh form (BA), retained antimicrobial activity against aquaculture pathogens, and preserved digestive enzyme production, hydrophobicity, aggregation, and antioxidant properties. Feeding L. rohita with BAFP (1 × 10⁹ cells g−1 feed) for 28 days significantly enhanced serum NBT, anti-protease, myeloperoxidase activities, albumin, and total protein while reducing glucose, SGOT, SGPT, CK-MB, and alkaline phosphatase. It also upregulated immune genes (TNF-α, C3, Lysozyme, IFN-ɣ) and downregulated iNOS in the head-kidney and liver. Challenge trials with Aeromonas hydrophila ATCC 7966, A. salmonicida subspecies salmonicida COFCAU_AS, and A. veronii COFCAU_AV1 showed significantly higher survival in BAFP-fed fish. These findings demonstrate that the optimized freeze-dried B. amyloliquefaciens formulation enhances probiotic stability and boosts host immunity, offering effective protection against bacterial pathogens in aquaculture.