摘要
在分离鉴定出东方蜜蜂微孢子虫(N.ceranae)的基础上,对蛋清源纳米硫化亚铁(nano-FeS)的合成方法进行优化并探究其对N.ceranae的体内外抑菌作用。从感染蜂群中分离致病菌,经形态学和分子生物学鉴定为N.ceranae。通过体外试验证明了nano-FeS对N.ceranae的抑菌作用及机制。通过体内试验证实了nano-FeS对N.ceranae病的治疗作用。结果显示,采用热溶剂法合成了nano-FeS,并通过正交试验筛选出最佳合成方法。流式细胞术和荧光染色试验证实,nano-FeS促进N.ceranae的凋亡坏死。nano-FeS处理后发现,N.ceranae细胞内发生铁富集,破坏谷胱甘肽和谷胱甘肽过氧化物酶抗氧化系统,并导致活性氧(ROS)积累,最终导致细胞膜脂质过氧化。体内试验表明,用nano-FeS饲喂蜜蜂后,感染N.ceranae的蜜蜂病死率降低,中肠内的孢子数量减少。利用转录组和荧光定量PCR揭示了nano-FeS对感染N.ceranae蜜蜂中肠基因表达的影响。结果表明,合成并优化了抗N.ceranae的nano-FeS的合成方法,平均粒径为75nm。本研究不仅提供了一种治疗蜜蜂微孢子虫感染的潜在药物,而且发现nano-FeS的抗真菌机制与铁死亡有关,nano-FeS对N.ceranae的抑菌作用与nano-FeS质量浓度呈正相关关系。
Based on the isolation and identification of Nosema ceranae(N.ceranae)in honeybees,this study optimized the synthesis method of nano-FeS derived from albumen and explored in vitro and in vivo antifungal effects against N.ceranae.Pathogens were isolated from infected honeybee colonies and identified as N.ceranae using morphological and molecular biology techniques.In vitro experiments were conducted to confirm the antifungal effects of nano-FeS against N.ceranae and elucidate the mechanism.In vivo experiments were carried out to validate the therapeutic effects of nano-FeS against N.ceranae infection.Nano-FeS was synthesized using the solvothermal method with an optimal scheme determined through orthogonal experiments,with an average particle size of 75nm.Flow cytometry and fluorescence staining experiments confirmed that nano-FeS induced apoptosis and necrosis in N.ceranae.After N.ceranae was exposed to nano-FeS,intracellular iron accumulation,disruption of the glutathione and glutathione peroxidase antioxidant system,and subsequent ROS accumulation were observed,ultimately leading to lipid peroxidation of cell membranes.In vivo experiments demonstrated reduced mortality and decreased spore counts in the midgut of honeybees fed with nano-FeS.Transcriptome analysis and qPCR revealed the impact of nano-FeS on gene expression in the N.ceranae infected honeybee midgut.This study presented a promising alternative antifungal agent for N.ceranae infection in honeybees and elucidated the antifungal mechanism of nano-FeS related to ferroptosis.Additionally,the study found a positive correlation between the mass concentration of nano-FeS and its antifungal effectiveness against N.ceranae.
作者
吴浩东
孙浩
陈乐
黄灿
程尚
曹礼静
王庆华
WU Haodong;SUN Hao;CHEN Le;HUANG Can;CHENG Shang;CAO Lijing;WANG Qinghua(College of Veterinary Medicine,Southwest University,Rongchang,Chongqing 402460,China;Husbandry Technology Popularization Master Station of Chongqing,Yubei,Chongqing 401121,China;Chongqing Rongchang District Vocational Education Center,Rongchang,Chongqing402460,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2024年第7期1418-1429,共12页
Chinese Journal of Veterinary Science
基金
重庆市教委重大基金资助项目(KJZD-M202200202)
中国农村专业技术协会重庆荣昌肉兔科技小院资助项目。
