Molecular and histopathological detection of Hepatozoon canis in red foxes (Vulpes vulpes) from Portugal
Molecular and histopathological detection of Hepatozoon canis in red foxes (Vulpes vulpes) from Portugal
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DOI：
10.1186/1756-3305-7-113
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发表时间：
2014-03-24
影响因子：
3.2
通讯作者：
Baneth, Gad
中科院分区：
文献类型：
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作者：
Cardoso, Luis;Cortes, Helder C. E.;Baneth, Gad
背景：犬肝虫是犬和野生犬科动物的原生动物蜱传病原体。肝虫属据报道，不同大陆的狐狸都会感染这种病毒，最近的研究主要使用聚合酶链式反应 (PCR) 来检测和表征感染物种。对红狐（Vulpes vulpes）进行调查可能有助于更好地了解犬媒介传播疾病的流行病学，包括由犬嗜血杆菌在家犬中引起的肝细胞病。本研究调查了肝虫属的流行情况。通过对来自葡萄牙不同地区的红狐的不同组织进行组织病理学和分子分析。方法：从葡萄牙北部、中部和南部八个地区的91只红狐中采集血液和组织，包括骨髓、心脏、后腿肌肉、空肠、肾、肝、肺、腘或腋窝淋巴结、脾和/或舌头。将组织用福尔马林固定、石蜡包埋、切割并用苏木精和伊红染色。聚合酶链式反应 (PCR) 扩增了与肝细胞动物属 (Hepatozoon spp) 18S rRNA 基因相似的 650 bp 片段。结果：通过PCR和测序，在所有采样区域的90只狐狸中，有68只（75.6%）检出犬肝原虫。组织病理学显示，47 只狐狸中，有 11 只 (23.4%) 的骨髓中和两只 (4.3%) 的脾脏中发现了犬嗜血杆菌的形状与狗相似 (p = 0.007)。所有 11 只狐狸经组织病理学检测呈阳性，骨髓和/或血液 PCR 检测也呈阳性。在来自相同 47 只狐狸的配对骨髓样本中，PCR 检测的阳性率 (83.0%) 显着高于组织病理学检查 (23.4%) 的阳性率 (p < 0.001)。犬肝孢子虫18S rRNA基因序列与GenBank中的序列一致性为98-99%。结论：犬肝孢子虫在葡萄牙北部、中部和南部的红狐群体中高度流行。通过组织病理学检测寄生虫的灵敏度明显低于 PCR 检测。红狐是家犬感染犬嗜血杆菌的假定宿主。
Background: Hepatozoon canis is a protozoan tick-borne pathogen of dogs and wild canids. Hepatozoon spp. have been reported to infect foxes in different continents and recent studies have mostly used the polymerase chain reaction (PCR) for the detection and characterization of the infecting species. Surveying red foxes (Vulpes vulpes) may contribute to better understanding the epidemiology of canine vector-borne diseases, including hepatozoonosis caused by H. canis in domestic dogs. The present study investigated the prevalence of Hepatozoon spp. by means of histopathology and molecular analysis of different tissues in red foxes from different parts of Portugal.Methods: Blood and tissues including bone marrow, heart, hind leg muscle, jejunum, kidney, liver, lung, popliteal or axillary lymph nodes, spleen and/or tongue were collected from 91 red foxes from eight districts in northern, central and southern Portugal. Tissues were formalin-fixed, paraffin-embedded, cut and stained with hematoxylin and eosin. Polymerase chain reaction (PCR) amplified a similar to 650 bp fragment of the 18S rRNA gene of Hepatozoon spp. and the DNA products were sequenced.Results: Hepatozoon canis was detected in 68 out of 90 foxes (75.6%) from all the sampled areas by PCR and sequencing. Histopathology revealed H. canis meronts similar in shape to those found in dogs in the bone marrow of 11 (23.4%) and in the spleen of two (4.3%) out of 47 foxes (p = 0.007). All the 11 foxes found positive by histopathology were also positive by PCR of bone marrow and/or blood. Positivity by PCR (83.0%) was significantly higher (p < 0.001) than by histopathological examination (23.4%) in paired bone marrow samples from the same 47 foxes. Sequences of the 18S rRNA gene of H. canis were 98-99% identical to those in GenBank.Conclusions: Hepatozoon canis was found to be highly prevalent in red fox populations from northern, central and southern Portugal. Detection of the parasite by histopathology was significantly less sensitive than by PCR. Red foxes are a presumptive reservoir of H. canis infection for domestic dogs.