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{"title":"Laboratory Growth and Genetic Manipulation of Eimeria tenella","authors":"Iván Pastor-Fernández, Elaine Pegg, Sarah E. Macdonald, Fiona M. Tomley, Damer P. Blake, Virginia Marugán-Hernández","doi":"10.1002/cpmc.81","DOIUrl":null,"url":null,"abstract":"<p><i>Eimeria</i> is a genus of apicomplexan parasites that contains a large number of species, most of which are absolutely host-specific. Seven species have been recognized to infect chickens. Infection of susceptible chickens results in an intestinal disease called coccidiosis, characterized by mucoid or hemorrhagic enteritis, which is associated with impaired feed conversion or mortality in severe cases. Intensive farming practices have increased the significance of coccidiosis since parasite transmission is favored by high-density housing of large numbers of susceptible chickens. Routine chemoprophylaxis and/or vaccination with live parasite vaccines provides effective control of <i>Eimeria</i>, although the emergence of drug resistance and the relative cost and production capacity of current vaccine lines can prove limiting. As pressure to reduce drug use in livestock production intensifies, novel vaccination strategies are needed. Development of effective protocols supporting genetic complementation of <i>Eimeria</i> species has until recently been hampered by their inability to replicate efficiently in vitro. Now, the availability of such protocols has raised the prospect of generating transgenic parasite lines that function as vaccine vectors to express and deliver heterologous antigens. For example, this technology has the potential to streamline the production of live anticoccidial vaccines through the generation of parasite lines that co-express immunoprotective antigens derived from multiple <i>Eimeria</i> species. In this paper we describe detailed protocols for genetic manipulation, laboratory growth, and in vivo propagation of <i>Eimeria tenella</i> parasites, which will encourage future work from other researchers to expand biological understanding of <i>Eimeria</i> through reverse genetics. © 2019 by John Wiley & Sons, Inc.</p>","PeriodicalId":39967,"journal":{"name":"Current Protocols in Microbiology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2019-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmc.81","citationCount":"23","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Microbiology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmc.81","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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Abstract
Eimeria is a genus of apicomplexan parasites that contains a large number of species, most of which are absolutely host-specific. Seven species have been recognized to infect chickens. Infection of susceptible chickens results in an intestinal disease called coccidiosis, characterized by mucoid or hemorrhagic enteritis, which is associated with impaired feed conversion or mortality in severe cases. Intensive farming practices have increased the significance of coccidiosis since parasite transmission is favored by high-density housing of large numbers of susceptible chickens. Routine chemoprophylaxis and/or vaccination with live parasite vaccines provides effective control of Eimeria , although the emergence of drug resistance and the relative cost and production capacity of current vaccine lines can prove limiting. As pressure to reduce drug use in livestock production intensifies, novel vaccination strategies are needed. Development of effective protocols supporting genetic complementation of Eimeria species has until recently been hampered by their inability to replicate efficiently in vitro. Now, the availability of such protocols has raised the prospect of generating transgenic parasite lines that function as vaccine vectors to express and deliver heterologous antigens. For example, this technology has the potential to streamline the production of live anticoccidial vaccines through the generation of parasite lines that co-express immunoprotective antigens derived from multiple Eimeria species. In this paper we describe detailed protocols for genetic manipulation, laboratory growth, and in vivo propagation of Eimeria tenella parasites, which will encourage future work from other researchers to expand biological understanding of Eimeria through reverse genetics. © 2019 by John Wiley & Sons, Inc.
柔嫩艾美耳球虫的实验室生长和基因操作
艾美耳球虫是顶复体寄生虫的一个属,包含大量的种类,其中大多数是绝对的宿主特异性。已经确认有7种细菌会感染鸡。易感鸡的感染导致一种称为球虫病的肠道疾病,其特征是粘液性或出血性肠炎,严重时与饲料转化率受损或死亡有关。集约化养殖方式增加了球虫病的重要性,因为高密度饲养大量易感鸡有利于寄生虫传播。常规化学预防和/或接种寄生虫活疫苗可有效控制艾美耳球虫,尽管耐药性的出现以及目前疫苗品系的相对成本和生产能力可能会受到限制。随着畜牧生产中减少药物使用的压力加剧,需要新的疫苗接种策略。直到最近,支持艾美球虫物种遗传互补的有效方案的发展一直受到其无法在体外有效复制的阻碍。现在,这种方案的可用性提高了产生转基因寄生虫系作为表达和传递异源抗原的疫苗载体的前景。例如,这项技术有可能通过产生共表达来自多个艾美耳球虫物种的免疫保护性抗原的寄生虫系来简化抗球虫活疫苗的生产。在本文中,我们详细描述了艾美耳球虫的遗传操作,实验室生长和体内繁殖的详细方案,这将鼓励其他研究人员通过反向遗传学来扩大艾美耳球虫的生物学理解。©2019 by John Wiley &儿子,Inc。
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