Department of Animal Parasitology, Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science, Hangzhou, 310021, China,
Parasitol Res. 2014 Mar;113(3):1085-90. doi: 10.1007/s00436-013-3744-1. Epub 2014 Jan 14.
An Eimeria intestinalis isolated from a rabbit in China was first identified by amplifying the 18S small subunit (SSU) ribosomal RNA gene. 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 × 10(3), 5 × 10(4), 5 × 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 × 10(4) and 5 × 10(5) oocysts. And one rabbit (25 %) inoculated with 5 × 10(5) oocysts died 15 days after the inoculation. In the immunogenicity assay, SPF rabbits in five groups (named B1, B2, B3, B4, and B5) were immunised with 5 × 10(1), 5 × 10(2), 5 × 10(3), 0, and 0 sporulated oocysts, respectively. All rabbits but the B5 group were challenged with 1 × 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.
一株从中国兔体内分离到的艾美耳球虫,首先通过扩增 18S 小亚基(SSU)核糖体 RNA 基因来鉴定。扩增片段的大小为 1521bp。该艾美耳球虫分离株的 18S SSU RNA 基因与来自法国和捷克共和国的艾美耳球虫分离株的序列同源性为 99%,覆盖率分别为 100%和 96%。然后,在无特定病原体(SPF)兔中评估该艾美耳球虫分离株的致病性和免疫原性。在致病性试验中,将 4 组 SPF 兔分别感染 5×10^3、5×10^4、5×10^5 和 0 个孢子化卵囊。感染 5×10^4 和 5×10^5 卵囊的兔出现腹泻、便秘、食欲不振和体重增长减少等临床症状。感染 5×10^5 卵囊的一只兔(25%)在接种后 15 天死亡。在免疫原性试验中,将 5 组 SPF 兔(命名为 B1、B2、B3、B4 和 B5)分别用 5×10^1、5×10^2、5×10^3、0 和 0 个孢子化卵囊免疫。除 B5 组外,所有兔子均用 1×10^6 个卵囊攻毒。攻毒后,B2 和 B3 组兔子无或仅有轻微临床症状。与对照组相比,B2 和 B3 组卵囊排出量分别减少了 69.6%和 84.5%。两组兔子的体重增长明显高于攻毒对照组。所有结果表明,该艾美耳球虫分离株在兔中具有低毒力但免疫原性。