贝氏隐孢子虫鸡胚培养方法研究  

Cryptosporidium baileyi oocysts cultured in chicken embryo

作　　者：黄磊[1] 张云[1] 张素梅[1] 菅复春[1] 宁长申[1] 张龙现[1] 

机构地区：[1]河南农业大学牧医工程学院,郑州450002

出　　处：《中国人兽共患病学报》2012年第3期205-208,共4页Chinese Journal of Zoonoses

基　　金：教育部博士点基金项目(No.20094105110003);国家自然科学基金项目(No.30871863)联合资助

摘　　要：目的建立能稳定获得大量贝氏隐孢子虫(Cryptosporidium baileyi)卵囊的鸡胚培养方法,应用于抗隐孢子虫药物筛选和C.baileyi体外传代培养研究。方法研究鸡胚培养温度、卵囊接种量、卵囊脱囊处理和卵囊回收时间对C.bai-leyi卵囊在鸡胚中增殖量的影响。结果 C.baileyi卵囊经尿囊腔接种9日龄鸡胚后在37℃、38℃、40℃条件下培养168h,各组鸡胚尿囊液中卵囊量差异不显著(P>0.05),但随着培养温度升高卵囊增殖量呈增加趋势。卵囊接种量分别为3.0×105个/枚和1.0×106个/枚,鸡胚40℃培养168h,尿囊液中卵囊增殖倍数差异不显著(P>0.05)。接种3.0×105个卵囊和等量卵囊脱囊后产物,鸡胚40℃培养168h,尿囊液中卵囊量差异不显著(P>0.05)。接种3.0×105个卵囊,鸡胚40℃培养168h,尿囊液中卵囊量显著高于接种后144h和接种后192h(P<0.05)。结论尿囊液收集时间是影响C.baileyi鸡胚培养卵囊获得量的重要因素,培养温度和接种量以40℃和3.0×105个/枚为宜,而卵囊脱囊处理是不必要的。The aim of the present study was to establish a the chicken embryo culture method acquiring a large number of Cryptosporidium baileyi oocysts for anti-Cryptosporidium drug screening and its vitro culture,in which some conditions containing the effect of temperature,inoculum dose,excystation and recovery time were considered. The 9-day-old chicken embryos were innoculated with 3×105 oocysts in allantoic cavity and incubated for a further 168 hours at 37℃,38℃ and 40℃,respectively,in which the number of reproduction of oocysts was not significant different（P0.05）.Innoculated with 3×105 oocysts the reproduction rates of oocysts was similar compared with innoculated with 1×106oocysts（P0.05） and innoculated with 3×105 oocysts treated by 0.75% synthetic sodium taurocholate the number of reproduction of oocysts was not significant different compared with innoculated with 3×105oocysts treated by 0.75% synthetic sodium taurocholate（P0.05）.But the number of reproduction of oocysts incubated for a further 168h was more than incubated for a further 144h and 192h（P0.05）.The conclusions were that recovery time was vital factor in collecting more C.baileyi oocysts.Oocysts excystation was unnecessary and it was suitable that chicken embryos innoculate with 3×105 oocysts at 40℃.

关 键 词：贝氏隐孢子虫 鸡胚 体外培养 

分 类 号：R382.3[医药卫生—医学寄生虫学]