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首页> 外文期刊>Parasitology >High-yield amplification of Cryptosporidium parvum in interferon gamma receptor knockout mice.
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High-yield amplification of Cryptosporidium parvum in interferon gamma receptor knockout mice.

机译：干扰素γ受体敲除小鼠中小隐孢子虫的高产扩增。

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To date, large-scale production of Cryptosporidium parvum oocysts has only been achieved by amplification in neonatal calves and sheep. Many laboratories currently depend on supplies from external sources and store oocysts for prolonged periods which results in progressive loss of viability. Six to 8-week-old interferon gamma receptor knockout (IFN gamma R-KO) mice on a C57BL/6 background were inoculated by gavage (2000 oocysts/animal). Fecal pellets were collected daily from 7 days post-infection (p.i.) up to 2 weeks p.i. Intestinal oocyst yield was assessed at days 11, 12 and 14 p.i. by homogenization of intestinal tissues. Ether extraction and one or more NaCl flotations were used to purify oocysts. Total recoveries averaged 2.6 x 10(6) oocysts/mouse from fecal material and 3.8 x 10(7) oocysts/mouse from intestinal tissues. Overall, 2.3 x 10(9) purified oocysts were obtained from 60 mice. Recovered oocysts were capable of sporulation and were shown to be infectious both in vitro and in vivo. Oocyst amplification was achieved in only 11-14 days with minimal expense. The simplicity of this method presents a practical alternative for the routine passage, maintenance and storage of C. parvum in biomedical laboratories.
机译：迄今为止，小隐隐孢子虫卵囊的大规模生产只能通过在新生小牛和绵羊中扩增来实现。当前，许多实验室依赖于外部来源的供应并将卵囊长时间存储，这导致生存能力逐渐丧失。通过管饲法（2000个卵囊/动物）接种在C57BL / 6背景上的六至八周大的干扰素γ受体敲除（IFNγR-KO）小鼠。从感染后（p.i.）到感染后7天至每天2周，每天收集粪便颗粒。在第11、12和14天评估肠卵囊产量。通过肠组织的均质化。乙醚萃取和一种或多种NaCl浮选法用于纯化卵囊。从粪便中回收的平均平均2.6 x 10（6）个卵囊/小鼠，从肠组织回收的平均平均回收率为3.8 x 10（7）卵囊/小鼠。总体而言，从60只小鼠中获得了2.3 x 10（9）个纯化的卵囊。回收的卵囊能够形成孢子，并且在体外和体内均具有传染性。仅花费11-14天即可完成卵囊扩增。该方法的简单性为生物医学实验室中的小球藻的常规通过，维护和储存提供了一种实用的选择。

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