Yu Yu, Huilin Guo, Li Liujia, Guan Guiquan, Zhao Qiping, Zhu Shunhai, Wang Jinwen, Zhao Fanghe, Dong Hui, Han Hongyu
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai, 200241, China.
College of Agriculture and Biological Science, Dali University, Dali, Yunnan, 671003, China.
Int J Parasitol Drugs Drug Resist. 2025 Sep 19;29:100615. doi: 10.1016/j.ijpddr.2025.100615.
Coccidiosis, which is primarily caused by Eimeria spp., poses a persistent challenge to poultry health and production worldwide. The emergence and spread of drug-resistant strains have significantly compromised the efficacy of anticoccidial therapies. We previously used RNA-seq to demonstrate that ribosomal protein L27 is differentially expressed in drug-sensitive and maduramicin-resistant strains of E. tenella (EtRPL27). In the present study, an RT-qPCR analysis showed that its expression is stage-specific, with the highest levels in sporozoites and second-generation merozoites. Immunofluorescence revealed both the cytoplasmic and partial surface localization of EtRPL27. Notably, EtRPL27 transcript levels were significantly elevated in a maduramicin-resistant strain relative to the drug-sensitive strain. Functional assays showed that anti-EtRPL27 antibodies inhibited the invasion of sporozoites, whereas the transgenic overexpression of EtRPL27 enhanced both the invasion efficiency and pathogenicity of E. tenella in chickens. Importantly, EtRPL27 overexpression reduced its sensitivity to maduramicin, evident as increased oocyst output and a higher anticoccidial index. Resistance classification based on standard indices confirmed moderate-to-high resistance levels in the EtRPL27-overexpressing strain. These findings demonstrate that EtRPL27 is closely associated with both virulence and maduramicin resistance in E. tenella, indicating its potential utility as a molecular marker and therapeutic target.
球虫病主要由艾美耳属(Eimeria spp.)引起,对全球家禽健康和生产构成持续挑战。耐药菌株的出现和传播严重损害了抗球虫疗法的疗效。我们之前利用RNA测序证明核糖体蛋白L27在柔嫩艾美耳球虫(EtRPL27)的药物敏感株和马杜霉素耐药株中差异表达。在本研究中,RT-qPCR分析表明其表达具有阶段特异性,在子孢子和第二代裂殖子中水平最高。免疫荧光显示EtRPL27在细胞质和部分表面定位。值得注意的是,与药物敏感株相比,马杜霉素耐药株中EtRPL27转录水平显著升高。功能试验表明,抗EtRPL27抗体抑制子孢子的入侵,而EtRPL27的转基因过表达增强了柔嫩艾美耳球虫在鸡体内的入侵效率和致病性。重要的是,EtRPL27过表达降低了其对马杜霉素的敏感性,表现为卵囊产量增加和抗球虫指数更高。基于标准指标的耐药性分类证实EtRPL27过表达菌株具有中到高耐药水平。这些发现表明EtRPL27与柔嫩艾美耳球虫的毒力和马杜霉素耐药性密切相关,表明其作为分子标记和治疗靶点的潜在用途。