Development of molecular diagnostic protocols for detecting three types of Entamoeba from diarrheal and asymptomatic pigs and environmental moist soils
Development of molecular diagnostic protocols for detecting three types of Entamoeba from diarrheal and asymptomatic pigs and environmental moist soils
复制标题
DOI：
10.1007/s00436-017-5483-1
复制
发表时间：
2017-07-01
影响因子：
2
通讯作者：
Shibahara, Tomoyuki
中科院分区：
文献类型：
--
作者：
Hirashima, Yoshimasa;Manchanayake, Tilusha;Shibahara, Tomoyuki
猪内阿米巴以及波列基内阿米巴1型和3型亚型（ST）近期被认为与猪的疾病暴发有关。然而，这些寄生虫在日本的分布情况以及农场潜在的感染源仍不清楚。在此，我们对日本鹿儿岛县一个有粪便异常的育肥/生长猪养殖场进行了检查，发现养殖场环境中存在寄生虫。对患有溃疡性结肠炎的猪的肠道组织进行检查，发现大量滋养体侵入了病变部位。我们在受感染猪的石蜡切片或粪便样本中鉴定出猪内阿米巴以及波列基内阿米巴ST1和ST3的单一感染和混合感染。通过聚合酶链反应（PCR）和测序，使用四套引物鉴定出两种内阿米巴亚型。在饮水源周围或水坑附近获取的潮湿土壤样本中检测到了寄生虫，这表明包囊可通过受污染的土壤传播。此外，我们在两个相邻农场没有腹泻症状的受检猪中发现了内阿米巴属寄生虫及其混合感染的证据。我们的研究结果建立了成功鉴定寄生虫的方法，包括存在多种感染的情况。
Entamoeba suis and Entamoeba polecki subtypes (ST) 1 and 3 have recently been implicated in disease outbreaks in pigs. However, the distributions of these parasites in Japan and the potential sources of infection on farms still remain unclear. Here, we examined a farm of fattening/growing pigs with abnormal feces in Kagoshima Prefecture, Japan, and found the presence of parasites in the farm environment. Examination of intestinal tissues from pigs presenting with ulcerative colitis revealed a large number of trophozoites that had invaded the lesions. We identified single and mixed infections of E. suis and E. polecki ST1 and ST3 in paraffin sections or fecal samples from affected pigs. Two subtypes of Entamoeba were identified using four primer sets by PCR and sequencing. The parasites were detected in moist soil samples obtained around the drinking water source or puddles, implicating transmission of cysts via contaminated soils. Additionally, we found evidence of Entamoeba spp. and coinfections in surveyed pigs without any diarrhea at two neighboring farms. Our results establish methods for successfully identification of parasites, including cases in which multiple infections are present.