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<p><i>Pseudocohnilembus persalinus</i> is a highly pathogenic scuticociliate that poses a serious threat to marine aquaculture by parasitizing the skin, gills, muscles, and brain tissues of cultured animals, often leading to high mortality. In this study, the efficacy of ozone/hydrogen peroxide (O<sub>3</sub>/H<sub>2</sub>O<sub>2</sub>) advanced oxidation technology for inactivating <i>P. persalinus</i> was systematically evaluated using light and scanning electron microscopy, cell viability assays (CCK-8), and molecular analyses. Electron spin resonance (ESR) spectroscopy was employed to characterize the reactive oxygen species (ROS) generated during treatment. Results showed that at an O<sub>3</sub> dosage of 10&#xa0;g/h, complete inactivation required 140&#xa0;s with O<sub>3</sub> alone, but only 90&#xa0;s with O<sub>3</sub>/H<sub>2</sub>O<sub>2</sub>. Morphological analyses revealed extensive cellular damage, including membrane shrinkage, rupture, and cytoplasmic leakage. ESR analysis confirmed the generation of highly oxidative ROS, including hydroxyl radicals (•OH), superoxide anions (•O<sub>2</sub>⁻), and singlet oxygen (<sup>1</sup>O<sub>2</sub>). The CCK-8 assay showed that cell viability in the O<sub>3</sub>/H<sub>2</sub>O<sub>2</sub> group decreased to (4.50 ± 1.16)% after 90&#xa0;s. Agarose gel electrophoresis indicated severe DNA fragmentation, suggesting irreversible genomic damage. Transcriptomic analysis revealed significant changes in genes related to cellular components (12 genes) and pathways associated with membrane dynamics, neural signaling, and organelle integrity, with differentially expressed genes such as ECM and integrins linked to structural disruption. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment highlighted steroid hormone biosynthesis, mitogen-activated protein kinase (MAPK), and mechanistic target of rapamycin (mTOR) signaling pathways, implicating apoptosis induction. Collectively, these findings demonstrate that the rapid and effective inactivation of <i>P. persalinus</i> by O<sub>3</sub>/H<sub>2</sub>O<sub>2</sub> is primarily mediated through ROS-induced disruption of cellular integrity and physiological functions. This work provides a mechanistic basis for applying O<sub>3</sub>/H<sub>2</sub>O<sub>2</sub> technology in the prevention and control of parasitic infections in aquaculture systems.</p> Graphical Abstract <p></p>

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Rapid and efficient inactivation of Pseudocohnilembus persalinus in aquaculture water using O3/H2O2 advanced oxidation technology

  • 期刊论文

  • 发表日期：

    2026年3月19日

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  • Yuwei Cui,
  • Jingfeng Jia,
  • Xiaojuan Shao,
  • Wulong Jiang,
  • Wenke Liu,
  • Hua Wang,
  • Ruijun Li,
  • Zhongming Huo

摘要

Pseudocohnilembus persalinus is a highly pathogenic scuticociliate that poses a serious threat to marine aquaculture by parasitizing the skin, gills, muscles, and brain tissues of cultured animals, often leading to high mortality. In this study, the efficacy of ozone/hydrogen peroxide (O3/H2O2) advanced oxidation technology for inactivating P. persalinus was systematically evaluated using light and scanning electron microscopy, cell viability assays (CCK-8), and molecular analyses. Electron spin resonance (ESR) spectroscopy was employed to characterize the reactive oxygen species (ROS) generated during treatment. Results showed that at an O3 dosage of 10 g/h, complete inactivation required 140 s with O3 alone, but only 90 s with O3/H2O2. Morphological analyses revealed extensive cellular damage, including membrane shrinkage, rupture, and cytoplasmic leakage. ESR analysis confirmed the generation of highly oxidative ROS, including hydroxyl radicals (•OH), superoxide anions (•O2⁻), and singlet oxygen (1O2). The CCK-8 assay showed that cell viability in the O3/H2O2 group decreased to (4.50 ± 1.16)% after 90 s. Agarose gel electrophoresis indicated severe DNA fragmentation, suggesting irreversible genomic damage. Transcriptomic analysis revealed significant changes in genes related to cellular components (12 genes) and pathways associated with membrane dynamics, neural signaling, and organelle integrity, with differentially expressed genes such as ECM and integrins linked to structural disruption. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment highlighted steroid hormone biosynthesis, mitogen-activated protein kinase (MAPK), and mechanistic target of rapamycin (mTOR) signaling pathways, implicating apoptosis induction. Collectively, these findings demonstrate that the rapid and effective inactivation of P. persalinus by O3/H2O2 is primarily mediated through ROS-induced disruption of cellular integrity and physiological functions. This work provides a mechanistic basis for applying O3/H2O2 technology in the prevention and control of parasitic infections in aquaculture systems.

Graphical Abstract