National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan; Department of Microbiology and Parasitology, Sher-e-Bangla Agricultural University, Sher-e-Bangla Nagar, Dhaka 1207, Bangladesh.
Department of Parasitology, Faculty of Veterinary Medicine, Selcuk University, 42042 Konya, Turkey.
Parasitol Int. 2020 Dec;79:102163. doi: 10.1016/j.parint.2020.102163. Epub 2020 Jun 24.
The studies on Cryptosporidium infections of animals in Turkey mostly rely on microscopic observation. Few data are available regarding the prevalence of Cryptosporidium genotypes and subtypes infection. The aim of this study is to analyse the detection of Cryptosporidium genotypes and subtypes from young ruminants. A total of 415 diarrheic fecal specimens from young ruminants were examined for the Cryptosporidium detection by use of nested PCR of the small subunit ribosomal RNA (SSU rRNA) gene and the highly polymorphic 60 kDa glycoprotein (gp60) gene followed by sequence analyses. The results of this study revealed that 25.6% (106 of 415) of the specimens were positive for Cryptosporidium spp. infection. We identified 27.4% (91/333), 19.4% (13/67), and 13.4% (2/15) of positivity in calves, lambs and goat kids, respectively. Genotyping of the SSU rRNA indicated that almost all positive specimens were of C. parvum, except for one calf which was of C. bovis. Sequence analysis of the gp60 gene revealed the most common zoonotic subtypes (IIa and IId) of C. parvum. We detected 11 subtypes (IIaA11G2R1, IIaA11G3R1, IIaA12G3R1, IIaA13G2R1, IIaA13G4R1, IIaA14G1R1, IIaA14G3R1, IIaA15G2R1, IIdA16G1, IIdA18G1, IIdA22G1); three of them (IIaA12G3R1, IIaA11G3R1 and IIaA13G4R1) was novel subtypes found in calves and lambs. Additionally, three subtypes (IIaA11G2R1, IIaA14G3R1, and IIdA16G1) were detected in young ruminants for the first time in Turkey. These results indicate the high infection of Cryptosporidium in Turkey and propose that young ruminants are likely a major reservoir of C. parvum and a potential source of zoonotic transmission.
本研究旨在分析幼反刍动物中隐孢子虫基因型和亚型的检测情况。总共使用巢式 PCR 检测了 415 份来自幼反刍动物的腹泻粪便标本的小亚基核糖体 RNA (SSU rRNA) 基因和高度多态性 60 kDa 糖蛋白 (gp60) 基因,以检测隐孢子虫。序列分析结果显示,415 份标本中有 25.6%(106/415)为隐孢子虫属感染阳性。我们在犊牛、羔羊和山羊幼仔中分别鉴定出 27.4%(91/333)、19.4%(13/67)和 13.4%(2/15)的阳性率。SSU rRNA 的基因分型表明,除了一头犊牛为牛隐孢子虫外,几乎所有阳性标本均为小隐孢子虫。gp60 基因序列分析显示,小隐孢子虫最常见的人畜共患病亚型(IIa 和 IId)。我们检测到 11 种亚型(IIaA11G2R1、IIaA11G3R1、IIaA12G3R1、IIaA13G2R1、IIaA13G4R1、IIaA14G1R1、IIaA14G3R1、IIaA15G2R1、IIdA16G1、IIdA18G1、IIdA22G1);其中 3 种(IIaA12G3R1、IIaA11G3R1 和 IIaA13G4R1)为在犊牛和羔羊中发现的新型亚型。此外,在土耳其,幼反刍动物中首次检测到 3 种亚型(IIaA11G2R1、IIaA14G3R1 和 IIdA16G1)。这些结果表明,土耳其隐孢子虫感染率较高,提示幼反刍动物可能是小隐孢子虫的主要宿主,也是人畜共患病传播的潜在来源。