PARASITOLOGY RESEARCH
·
2023
·Vol. 123
·期 1
·1-6
·
IF 2
Wang Bingxiang,Zhao Ningning,Sun Lingyu,Tan Qianqian,Xiao Qianqian,Chen Junpeng,Li Jinxuan,Zhang Xiao,Zhao Xiaomin
Difficulties of in vitro culture and genetic manipulation of Eimeria tenella have hindered the screening of virulence factors in this parasite. In this study, the E. tenella rhoptry protein 30 (EtROP30) was expressed in Toxoplasma gondii (RH∆Ku80-EtROP30), and its effect on the proliferation and virulence of parasites was investigated. The results revealed that the expression of EtROP30 had no impact on the invasion and egress processes. However, the RH∆Ku80-EtROP30 strain formed larger plaques compared to the RH∆Ku80, indicating that the EtROP30 expression promotes T. gondii proliferation. Furthermore, the RH∆Ku80-EtROP30 strain exhibited greater pathogenicity, resulting in earlier mortality and shorter overall survival time compared to RH∆Ku80. These results imply that EtROP30 expression facilitates parasite intracellular proliferation and virulence in mice, suggesting that EtROP30 might be a candidate virulence factor of E. tenella.
研究领域:
寄生虫学
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分子生物学
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传染病学
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DOI
PubMed
高影响因子
PARASITOLOGY RESEARCH
·
2022
·Vol. 121
·期 5
·1507-1516
·
IF 2.383
Bingxiang Wang,Ningning Zhao,Yakun Wang,Lingyu Sun,Hongmei Li,Zhang Xiao,Zhao Xiaomin
Rhoptry proteins (ROPs), secreted by specific rhoptry organelles of apicomplexan parasites, are determinants of parasite pathogenesis and sources of vaccine candidates. Twenty-eight ROPs of Eimeria tenella have been predicted by genomic approaches, and in the present study, E. tenella rhoptry protein 30 (EtROP30) was characterized. Subcellular localizations of EtROP30 in sporozoites and merozoites were in the apical complex and rhoptry-like bulb, suggesting that EtROP30 is a member of ROPs in E. tenella . Sequence analysis showed that EtROP30 contained an N-terminal secretory signal, a protein kinase domain with eight E. tenella -specific rhoptry kinase 1 subfamily (ROPK-Eten1) motifs, and a C-terminal nuclear localization sequence (NLS), making EtROP30 the only ROP that contains both a secretory signal and an NLS in E. tenella . Subsequent experiments showed that EtROP30 was a secreted protein in the sporozoite stage, relying on NLS for migration to the host nucleus. In addition, EtROP30 showed significantly higher expression levels in the parasite merozoite stage, indicating that EtROP30 plays a critical role during parasite reinvasion and development and may be a viable option as a vaccine candidate for anti-parasitic infection. The immunization protection efficacies of EtROP30 were evaluated. Significant improvements in mean body weight gain, reduction of cecum lesion score, and number of oocysts excreted were observed, indicating that EtROP30 has good immunogenicity against E. tenella . In the present study, a ROP of E. tenella with secretory and nuclear localization characteristics has been identified, and proved to be an effective vaccine candidate against this parasite.
研究领域:
寄生虫学
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免疫学
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分子生物学
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疫苗学
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DOI
PubMed
高影响因子
International Journal for Parasitology-Drugs and Drug Resistance
·
2023
·
IF 4
Hongtao Zhang,Lei Zhang,Ganglin Ren,Hongbin Si,Xingju Song,Xianyong Liu,Xun Suo,Dandan Hu
Worldwide distributed coccidiosis is caused by infection of both Eimeria species and Cystoisospora in the host intestine and causes huge economic losses to the livestock industry, especially the poultry industry. The control of such diseases relies mainly on chemoprophylaxis with anticoccidials, which has led to a very common drug resistance in this field. However, the genetic mechanisms underlying resistance to many anticoccidial drugs remain unknown. In this study, strains of E. tenella resistant to 250 mg/kg monensin were generated and characterized. Forward genetic approaches based on pooled genome sequencing, including experimental evolution and linkage group selection, were used to locate candidate targets responsible for resistance to monensin and diclazuril in E. tenella . A total of 16 nonsynonymous mutants in protein-coding genes were identified in monensin-resistant strains, and two genomic regions with strong selection signals were also detected in diclazuril-resistant strains. Our study reveals the genetic characterization of the experimental evolution and linkage group selection in Eimeria species, and also provides important information that contributes to the understanding of the molecular mechanism of drug resistance in coccidia.
研究领域:
寄生虫学
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遗传学
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兽医学
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DOI
PubMed
高影响因子
International Journal for Parasitology-Drugs and Drug Resistance
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2025
·
IF 3.4
Yu Yu,Guo Huilin,Liujia Li,Guiquan Guan,Qiping Zhao,Shunhai Zhu,Jinwen Wang,Fanghe Zhao,Hui Dong,Hongyu Han
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 ( Et RPL27). 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 Et RPL27. Notably, Et RPL27 transcript levels were significantly elevated in a maduramicin-resistant strain relative to the drug-sensitive strain. Functional assays showed that anti- Et RPL27 antibodies inhibited the invasion of sporozoites, whereas the transgenic overexpression of Et RPL27 enhanced both the invasion efficiency and pathogenicity of E. tenella in chickens. Importantly, Et RPL27 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 Et RPL27-overexpressing strain. These findings demonstrate that Et RPL27 is closely associated with both virulence and maduramicin resistance in E. tenella, indicating its potential utility as a molecular marker and therapeutic target.
研究领域:
寄生虫学
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兽医学
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分子生物学
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抗菌素耐药性
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家禽健康
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产品关联
DOI
PubMed
高影响因子
VETERINARY RESEARCH
·
2025
·Vol. 56
·期 1
·1-13
·
IF 3.5
Wang Zengbao,Li Taifeng,Jiang Yingying,Wang Xue,Li Hongmei,Zhao Xiaomin,Zhang Xiao,Zhao Ningning
Serpin protease inhibitors (SERPIN) in protozoa play crucial roles in various biological processes, including the invasion of host cells. However, the precise roles and molecular mechanisms underlying SERPIN-mediated invasion of parasite remain poorly understand. In this study, we provide evidence that surface-expressed Eimeria tenella SERPIN1 (EtSERPIN1) on sporozoites is involved in adhesion and invasion processes. To elucidate the molecular target responsible for mediating EtSERPIN1-induced invasion, we utilized GST pull-down and yeast two-hybrid assays to identify host cell membrane proteins interacting with EtSERPIN1. Our findings revealed an interaction between EtSERPIN1 and a membrane protein called annexin A2 (ANXA2). Furthermore, recombinant GgANXA2 was shown to bind to the sporozoite surface. Pre-treatment with anti-GgANXA2-specific antibody or recombinant GgANXA2 protein significantly inhibited EtSERPIN1 binding to host cells. Pre-incubation with recombinant GgANXA2 also reduced sporozoite infection in both DF-1 cells and chickens. These results suggest that the interaction between EtSERPIN1 and GgANXA2 plays a critical role in both adhesion and invasion processes of E. tenella sporozoites. Finally, we investigated the impact of recombinant GgANXA2 and EtSERPIN1 proteins on E. tenella infection. Our results demonstrate that incubation with GgANXA2 protein significantly attenuated sporozoite infectivity, as evidenced by a significant reduction in parasite burden within the chicken cecum. Immunization with recombinant EtSERPIN1 exhibited potent anti- E. tenella activity, with higher body weight gains, lower cecal lesions and oocyst output, as well as elevated levels of cecal mucosa antibodies. These findings suggest that targeting GgANXA2 through EtSERPIN1 mediates adhesion and invasion processes of E. tenella , highlighting its potential as a novel therapeutic target.
研究领域:
寄生虫学
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分子生物学
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兽医学
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产品关联
DOI
PubMed
高影响因子
Animals
·
2025
·Vol. 16
·期 1
·67
·
IF 2.7
Liu-Shu Jia,Qing-Jie Wang,Shun-Hai Zhu,Qi-Ping Zhao,Yu Yu,Hong-Yu Han,Hui Dong
Simple SummarySodium hypochlorite (NaClO) is widely used for oocyst purification, yet the effect of NaClO on oocyst proteomic changes has not been reported. This study investigated the structural and proteomic alterations inEimeria tenellaunsporulated oocysts induced by NaClO treatment. Transmission electron microscopy revealed that NaClO disrupts the oocyst wall bilayer by removing the outer layer and inducing thickening of the inner layer. Label-free quantitative proteomics identified 1344 differentially expressed proteins (DEPs) between the NaClO-treated (Et-T) and the untreated (Et-C) unsporulated oocysts. Functional analysis showed that DEPs are primarily associated with oocyst wall biosynthesis, stress response pathways, outer wall formation, and structural integrity. These findings provide critical insights into the molecular architecture of the oocyst wall and establish a foundation for elucidating the mechanisms underlying its biosynthesis and environmental resilience.Sodium hypochlorite (NaClO) is widely used to purify oocysts in laboratories. While previous studies have extensively examined its effects on oocyst viability, pathogenicity, and sporulation rate, the impact of NaClO treatment on proteomic profiles remains uncharacterized. Transmission electron microscopy was used in the present study to characterize structural changes in unsporulated oocyst walls ofEimeria tenellatreated with NaClO. The results indicated that NaClO treatment destroyed the bilayer wall of unsporulated oocysts, stripping away the outer wall and making the inner layer thicker. Label-free quantitative proteomics was employed to identify differentially expressed proteins (DEPs) in NaClO-treated (Et-T) and untreated (Et-C) unsporulated oocysts. Among 2422 identified proteins, 1345 were differentially expressed, with 1210 upregulated and 134 downregulated in Et-T vs. Et-C. Functional analysis revealed that upregulated proteins are predominantly associated with oocyst wall biosynthesis and cellular stress responses, whereas downregulated proteins are involved in outer wall assembly and structural integrity. Notably, 12 proteins—including 9 hypothetical proteins, acid phosphatase, adenylate cyclase, and microneme protein 2—were exclusively detected in the Et-C, indicating their potential essentiality in outer wall formation. These findings reveal the structure and protein composition of the oocyst wall ofE. tenella, supporting research on its biosynthesis and environmental resilience.
研究领域:
寄生虫学
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蛋白质组学
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兽医学
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分子生物学
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微生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF VETERINARY MEDICAL SCIENCE
·
2024
·
IF 1.1
Nianyuan CHEN,Qingxiu CAI,Shujing WANG,Qingyang SONG,Ying XIE,Huijuan SHI,Hongmei LI,Xiaomin ZHAO,Ningning ZHAO,Xiao ZHANG
Protozoan parasites such as Eimeria tenella and Toxoplasma gondii pose significant health challenges in livestock and humans. The limited treatment options and rising drug resistance underscore the urgent need for new therapies. This study investigates myrcene, a monoterpene hydrocarbon classified for its antiprotozoal potential against E. tenella and T. gondii infections. Initially, we examined its effect on the sporulation process of E. tenella oocysts in vitro and its anti-E. tenella activity in vivo. Myrcene significantly reduced the sporulation rate of E. tenella oocysts at 3 and 4 mg/kg. In vivo experiments demonstrated that treatment with 4 mg/kg myrcene significantly reduced E. tenella load and oocyst output, as well as cecal lesion and weight loss caused by E. tenella infection, showing moderate anti-E. tenella activity, with an Anticoccidial Index (ACI) of 161.4. Furthermore, we investigated the anti-T. gondii activity of myrcene both in vitro and in vivo. In vitro studies showed that treatment with myrcene effectively inhibited the invasion rate and intracellular proliferation ability of T. gondii tachyzoite in DF-1 cells in a dose-dependent manner. In vivo administration prolonged the survival time in T. gondii-infected mice, suggesting notable protective effects. Additionally, it mitigated T. gondii-induced hepatosplenic toxicity by reducing parasite load in the liver and spleen, and ameliorating liver function as evidenced by decreased serum transaminase levels. In conclusion, the findings demonstrate promising anti-E. tenella and anti-T. gondii activity exhibited by myrcene warranting further exploration into its mechanisms and potential therapeutic applications.
研究领域:
兽医学
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寄生虫学
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药理学
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DOI
PubMed
高影响因子
JOURNAL OF APPLIED POULTRY RESEARCH
·
2026
·
IF 2
Tiantao Li,Chenzhong Fei,Xiaoyang Wang,Chunmei Wang,Wen Zhou,Ruihua Zhang,Keyu Zhang
Summary Prevention and control of coccidiosis remain a top priority for the poultry industry. Due to the increasing prevalence of resistance to chemical anticoccidial drugs and associated residue concerns in meat, eggs, and the environment, natural products have become an important alternative and complementary strategy for managing coccidiosis in poultry. However, research on the effects of varying proportions of herbal prescriptions on coccidiosis remains limited. In this study, different formulations were composed of Dichroa febrifuga Lour. (DFL), Artemisia annua L . (AAL), Acalypha australis L . (ACA), Pteris multifida Poir. (PMP) and Agrimonia pilosa Ledeb . (APL). Following Eimeria tenella infection, the effects of different formula compositions and extraction methods on coccidiosis resistance in broilers were evaluated. The results showed that, compared with the infection control group, most herbal formulations improved the fecal score, lesion score, and oocyst index in E. tenella -infected broilers. However, the anticoccidial efficacy index (ACI) of all formulations was moderate or limited. Based on the weight ratio, the optimal formulation was determined as DFL:ACA:PMP:APL = 4:1:2:2.
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DOI
高影响因子
VETERINARY PARASITOLOGY
·
2020
·
IF 2.157
Ruonan Bo,Xingru Dai,Jie Huang,Simin Wei,Mingjiang Liu,Jingui Li
Decoquinate (DQ) is used for prophylaxis against coccidian infections within the digestive tract of chickens, but DQ is extremely insoluble in water. Hence, improving the water solubility of DQ is extremely important. First, decoquinate nanoliposomes (DQNLs) were prepared by the thin-film dispersion–ultrasonic method. The preparation conditions of DQNLs were optimized using the orthogonal test. The optimal preparation conditions of DQNLs were: a ratio of egg-yolk lecithin:drug ( w/w ) of 10:1, ratio of egg-yolk lecithin:cholesterol ( w/w ) of 5:1, rotary-evaporation temperature of 50 ℃, and ultrasound duration of 15 min. The encapsulation efficiency of DQNLs prepared under these conditions reached 99.24 % and drug loading was 5.67 %. The characterization of optimized DQNLs was also done. Transmission electron microscopy of DQNLs showed that they had the characteristic structure of liposomes. The mean particle size was 115.6 nm. The polydispersity index was 0.175. The zeta potential was −39.1 mV. The stability of DQNLs was high upon storage at 4 ℃. In vivo studies demonstrated that the lower dose (5 mg/L) of DQNLs in drinking water obtained the similar anticoccidial efficacy to that of 40 mg/kg DQ in feed against diclazuril-resistance Eimeria tenella isolate. The in vitro inhibitory effect of DQNLs on the sporulation of Eimeria tenella oocysts was dose-dependent. Therefore, the anticoccidial efficacy of DQ was enhanced significantly after being encapsulated into nanoliposomes.
研究领域:
兽医学
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药学
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动物健康
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产品关联
DOI
PubMed
高影响因子
Pharmaceutics
·
2022
·Vol. 14
·期 7
·1330
·
IF 6.525
Yan Zhang,Runan Zuo,Xinhao Song,Jiahao Gong,Junqi Wang,Mengjuan Lin,Fengzhu Yang,Xingxing Cheng,Xiuge Gao,Lin Peng,Hui Ji,Xia Chen,Shanxiang Jiang,Dawei Guo
Maduramicin ammonium (MAD) is one of the most frequently used anticoccidial agents in broiler chickens. However, the high toxicity and low solubility of MAD limit its clinical application. In this study, MAD-loaded nanostructured lipid carriers (MAD–NLCs) were prepared to overcome the defects of MAD by using highly soluble nanostructured lipid carriers (NLCs). The formulation was optimized via a three-level, three-factor Box–Behnken response surface method. Then, the optimal MAD–NLCs were evaluated according to their hydrodynamic diameter (HD), zeta potential (ZP), crystal structure, encapsulation efficiency (EE), drug loading (DL), in vitro release, and anticoccidial effect. The optimal MAD–NLCs had an HD of 153.6 ± 3.044 nm and a ZP of −41.4 ± 1.10 mV. The X-ray diffraction and Fourier-transform infrared spectroscopy results indicated that the MAD was encapsulated in the NLCs in an amorphous state. The EE and DL were 90.49 ± 1.05% and 2.34 ± 0.04%, respectively, which indicated that the MAD was efficiently encapsulated in the NLCs. In the in vitro study, the MAD–NLCs demonstrated a slow and sustained drug release behavior. Notably, MAD–NLCs had an excellent anticoccidial effect againstEimeria tenellain broiler chickens. In summary, MAD–NLCs have huge potential to form a new preparation administered via drinking water with a powerful anticoccidial effect.Keywords:maduramicin ammonium;nanostructured lipid carriers;Box–Behnken design;anticoccidial effect
研究领域:
兽医学
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药剂技术
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动物健康
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产品关联
DOI
PubMed
高影响因子
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