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A low-virulence Eimeria intestinalis isolate from rabbit (Oryctolagm cuniculus) in China: molecular identification, pathogenicity, and immunogenicity

机译:中国兔子低毒力肠球菌艾美尔球虫分离株:分子鉴定,致病性和免疫原性

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An Eimeria intestinalis isolated from a rabbit in China was first identified by amplifying the 18S small subunit (SSU) ribosomal RNA ge,ne. The size of the amplified fragment was 1521 bp. The 18S SSU RNA gene of the E. intestinalis isolate shared 99 % sequence identity with E. intestinalis isolates from France and the Czech Republic, with 100 and 96 % coverage, respectively. Then, the pathogenicity and immunogenicity of the E. intestinalis isolate were evaluated in specific pathogen free (SPF) rabbits. In the pathogenicity assay, SPF rabbits in four groups were infected with 5 x 10~3, 5 x 10~4, 5 x 10~5, and 0 sporulated oocysts, respectively. Clinical signs including diarrhoea, constipation, loss of appetite, and reduction of body weight gain were observed in rabbits inoculated with 5 x 10~4 and 5x 10~5 oocysts. And one rabbit (25 %) inoculated with 5x10~5 oocysts died 15 days after the inoculation. In the immunogenicity assay, SPF rabbits in five groups (named Bl, B2, B3, B4, and B5) were immunised with 5 x 10~1, 5 x 10~2, 5 x 10~3, 0, and 0 sporulated oocysts, respectively. All rabbits but the B5 group were challenged with 1 x 10~6 oocysts. After the challenge, no or slight clinical signs were seen in rabbits of the B2 and B3 groups. Compared with the control, a 69.6 and 84.5 % reduction of oocyst output was observed in the B2 and B3 groups, respectively. The body weight gain of the two groups was obviously higher than that of the challenge control group. All the results show that the E. intestinalis isolate has low virulence but immunogenicity in rabbit.
机译:首先通过扩增18S小亚基(SSU)核糖体RNA ge,ne鉴定了从中国兔体内分离出的艾美球虫小肠。扩增片段的大小为1521bp。大肠埃希菌分离株的18S SSU RNA基因与法国和捷克共和国的大肠埃希菌分离株具有99%的序列同一性,覆盖率分别为100%和96%。然后,在无特定病原体(SPF)的兔子中评估肠小肠埃希菌分离株的致病性和免疫原性。在致病性测定中,四组SPF兔分别感染了5 x 10〜3、5 x 10〜4、5 x 10〜5和0个带孢子的卵囊。在接种5 x 10〜4和5x 10〜5卵囊的兔子中观察到腹泻,便秘,食欲不振和体重增加减少的临床体征。接种15x 5个卵囊的兔子(25%)死亡。在免疫原性测定中,用5 x 10〜1、5 x 10〜2、5 x 10〜3、0和0个带孢子的卵囊免疫五组(分别称为B1,B2,B3,B4和B5)的SPF兔。 , 分别。除B5组外,所有兔子均受到1 x 10〜6个卵囊的攻击。攻击后,在B2和B3组的兔子中未观察到或仅有轻微的临床体征。与对照组相比,B2和B3组的卵囊产量分别降低了69.6和84.5%。两组的体重增加明显高于挑战对照组。所有结果表明,肠内大肠杆菌分离物在兔中具有低毒力但具有免疫原性。

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