Department of Parasitology, Harbin Medical University, Harbin, Heilongjiang, China.
PLoS One. 2012;7(2):e31262. doi: 10.1371/journal.pone.0031262. Epub 2012 Feb 17.
Cryptosporidium and Giardia are the two important zoonotic pathogens causing diarrhea of humans and animals worldwide. Considering the human cryptosporidiosis outbreak and sporadic cases caused by C. cuniculus, the important public health significance of G. duodenalis and little obtained information regarding rabbit infected with Cryptosporidium and Giardia in China, the aim of this study is to determine the prevalence and molecularly characterize Cryptosporidium and Giardia in rabbits in Heilongjiang Province, China.
METHODOLOGY/PRINCIPAL FINDINGS: 378 fecal samples were obtained from rabbits in Heilongjiang Province. Cryptosporidium oocysts and Giardia cysts were detected using Sheather's sugar flotation technique and Lugol's iodine stain method, respectively. The infection rates of Cryptosporidium and Giardia were 2.38% (9/378) and 7.41% (28/378), respectively. Genotyping of Cryptosporidium spp. was done by DNA sequencing of the small subunit rRNA (SSU rRNA) gene and all the nine isolates were identified as Cryptosporidium cuniculus. The nine isolates were further subtyped using the 60-kDa glycoprotein (gp60) gene and two subtypes were detected, including VbA32 (n = 3) and a new subtype VbA21 (n = 6). G. duodenalis genotypes and subtypes were identified by sequence analysis of the triosephosphate isomerase (TPI) gene. The assemblage B (belonging to eight different subtypes B-I to B-VIII) was found in 28 G. duodenalis-positive samples.
CONCLUSIONS/SIGNIFICANCE: The rabbits have been infected with Cryptosporidium and Giardia in Heilongjiang Province. The results show that the rabbits pose a threat to human health in the studied areas. Genotypes and subgenotypes of C. cuniculus and G. duodenalis in this study might present the endemic genetic characterization of population structure of the two parasites.
隐孢子虫和贾第鞭毛虫是两种重要的人畜共患病原体,可导致全球人类和动物腹泻。鉴于人类隐孢子虫病爆发和由 C. cuniculus 引起的散发病例,以及在中国关于感染隐孢子虫和贾第鞭毛虫的兔的重要公共卫生意义知之甚少,本研究旨在确定中国黑龙江省兔感染隐孢子虫和贾第鞭毛虫的流行情况,并对其进行分子特征分析。
方法/主要发现:从黑龙江省采集了 378 份兔粪便样本。使用 Sheather's 糖漂浮技术和 Lugol's 碘染色法检测隐孢子虫卵囊和贾第鞭毛虫包囊。隐孢子虫和贾第鞭毛虫的感染率分别为 2.38%(9/378)和 7.41%(28/378)。通过小亚单位 rRNA(SSU rRNA)基因的 DNA 测序对隐孢子虫进行基因分型,9 株分离株均鉴定为兔隐孢子虫。进一步通过 60-kDa 糖蛋白(gp60)基因对 9 株分离株进行亚分型,共检测到 2 种亚型,包括 VbA32(n = 3)和新亚型 VbA21(n = 6)。通过三磷酸甘油醛异构酶(TPI)基因序列分析鉴定贾第鞭毛虫基因型和亚型。28 份 G. duodenalis 阳性样本中发现了 B 组(属于 8 种不同的 B-I 至 B-VIII 亚型)。
结论/意义:黑龙江省的兔已感染隐孢子虫和贾第鞭毛虫。结果表明,在研究地区,兔对人类健康构成威胁。本研究中 C. cuniculus 和 G. duodenalis 的基因型和亚基因型可能呈现出两种寄生虫种群结构的地方遗传特征。