大黄鱼源贪食迈阿密虫(Miamiensis avidus)的分离鉴定、体外培养及药物敏感性研究

    Isolation, identification, and in vitro characterization of Miamiensis avidus from large yellow croaker(Larimichthys crocea

    • 摘要:
      背景 大黄鱼(Larimichthys crocea)作为中国特色海水养殖鱼种,具有重要经济价值和产业地位,已成为中国海水养殖产量较高的鱼类之一。然而,随着集约化养殖的快速发展,多种因素导致养殖环境恶化,造成病害频发,对大黄鱼养殖业造成重大经济损失。其中,以盾纤毛虫病为代表的寄生虫病近年来日益严重。
      目的 本研究旨在分离并鉴定感染大黄鱼的盾纤毛虫种类,研究其病原学特征,以期为盾纤毛虫病防控提供理论基础。
      方法 本研究从宁德海区某大黄鱼养殖渔排采集锥体虫与盾纤毛虫共感染的鱼体样品,成功分离、培养并鉴定盾纤毛虫种类,建立稳定的体外培养体系。利用体外培养体系,评估盾纤毛虫在不同温度(0、10、20、25、30 °C)、pH(5.5、6.5、7.4、8.5、9.0)及盐度(0、10、20、30)等培养条件下的增殖特性。同时,开展体外药物敏感性实验,评估常用水产抗虫药物的杀虫效果。
      结果 形态学观察结合cox1和18S rRNA基因系统发育分析表明,本研究分离的盾纤毛虫为贪食迈阿密虫(Miamiensis avidus)。环境耐受性实验结果显示,虫体在温度为0 °C条件下12 h内全部死亡,在温度为30 °C条件下种群密度逐渐下降;温度为10 °C时仍可增殖但显著低于对照组(20 °C),而在温度为25 °C时与对照组差异不显著,提示该虫对高温(> 25 °C)较为敏感。盐度实验表明,虫体在淡水中无法存活,在盐度10条件下可增殖但密度显著低于对照组(盐度30),而盐度20条件下与对照组无显著差异,表明其具备较广的盐度耐受性。pH实验结果表明,虫体在pH 5.5 和pH 9.0下仍能增殖,但密度显著低于对照组(pH 7.4);在pH 6.5和pH 8.5下与对照组差异不显著,提示其具有较强的pH 适应性。体外药物敏感性实验显示,香芹酚在低剂量(100 mg/kg)即可显著杀灭贪食迈阿密虫。
      结论 本研究首次从中鱼阶段的大黄鱼中分离出盾纤毛虫,并鉴定其为贪食迈阿密虫,系统阐明了环境因子对其增殖的影响,并在常用水产药物中筛选出高效抗虫药物香芹酚。这些研究结果不仅有助于理解大黄鱼源贪食迈阿密虫的生物学特性,也为盾纤毛虫病的科学防控提供了理论基础。

       

      Abstract:
      Background The large yellow croaker (Larimichthys crocea) is a commercially valuable fish in China, widely farmed in marine aquaculture. However, rapid growth in intensive farming has led to poorer water quality and higher stocking densities, causing frequent disease outbreaks and economic losses. Notably, ciliate protozoan infections, especially scuticociliatosis, pose a significant threat.
      Objective This study aimed to isolate and identify the ciliate causing scuticociliatosis in large yellow croaker and to understand its pathogenic and biological traits, providing a basis for disease prevention and control.
      Methods In April 2025, fish co-infected with trypanosomes and scuticociliates were collected from a net-cage farm in Ningde, China. The parasites were isolated, cultured, and identified via morphological and molecular analysis. A stable culture system was developed to study the effects of temperature, pH, and salinity on ciliate growth. Drug sensitivity tests were also performed to evaluate common aquaculture anti-parasitics.
      Results Morphological and phylogenetic analyses confirmed the ciliate as Miamiensis avidus. Temperature tests showed high sensitivity: all cells died at 0°C within 12 hours, and population decreased at 30°C. Growth at 10°C was lower than at 20°C, with no significant difference at 25°C, indicating susceptibility to temperatures above 25 °C. Salinity tests revealed all cells died at salinity 0 within 12 hours, while growth at salinity 10 was lower than at salinity 30, with no significant difference at salinity 20, indicating broad salinity tolerance. The parasite could grow at pH 5.5 and pH 9.0, though less than at pH 7.4, with no significant difference at pH 6.5 and pH 8.5, showing strong pH adaptability. Drug sensitivity tests showed that carvacrol effectively eliminated M. avidus at a low dose (100 mg·kg−1), indicating its potential as a low-toxicity treatment.
      Conclusion The study confirms M. avidus as the cause of scuticociliatosis in sub-adult large yellow croaker, explores environmental factors affecting parasite growth, and identifies an effective low-dose treatment. These insights improve our understanding of M. avidus biology and support sustainable scuticociliatosis management in large yellow croaker aquaculture.

       

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