Hepatovirus infections in juvenile seals from the North Sea.

Monica Mirolo, Bianca Kühl, Melvin Daniel Roji, Ana Rubio-García, Valéria Andrade Lima, Christina Puff, Byron Martina, Andreas Beineke, Peter Wohlsein, Wolfgang Baumgärtner, Martin Ludlow, Albert Osterhaus
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Abstract

The discovery of several novel hepatovirus species in marine and terrestrial mammals has expanded the recognised members of the genus Hepatovirus and has provided better understanding on the evolutionary origins of human hepatovirus A (HAV). Using high throughput sequencing we detected a seal hepatovirus (SealHAV_NL/PV/21), in liver tissue of a deceased harbor seal (Phoca vitulina) originating from the Dutch North Sea coast. RT-PCR screening of liver samples of 88 harbor seals and 12 grey seals (Halichoerus grypus) from the same region identified seal hepatovirus in nine juvenile harbor seals in which minor sequence variation was observed in the VP1 gene. Whole-genome sequence analysis showed that SealHAV_NL/PV/21 displayed 95.6% nucleotide indentity to New England seal hepatovirus but had a 5'-UTR which contained additional 51 bp. Phylogenetic analysis showed that seal hepatoviruses clustered in a monophyletic group separate from other hepatovirus species that have been identified in terrestrial mammals. Assessment of seal hepatovirus RNA loads in organs of all infected animals showed that the liver had the highest number of RNA copies with up to 107 RNA copies per mg of tissue. Seal hepatovirus RNA was readily detected by in situ hybridization in hepatocytes in the liver but was not associated with pathological lesions. Serological screening of 90 contemporary seal sera using a HAV-based ELISA showed the presence of hepatovirus antibodies in 14 harbor seals and one juvenile grey seal. These findings collectively show that seal hepatovirus is enzootic among seals of the North Sea, causing quiescent infections in young animals.

北海幼海豹的肝病毒感染。
在海洋和陆地哺乳动物中发现了几种新的肝病毒,扩大了肝病毒属的已知成员,并为人类肝病毒A (HAV)的进化起源提供了更好的理解。采用高通量测序技术,在荷兰北海海岸一只死亡的斑海豹(Phoca vitulina)的肝脏组织中检测到一种海豹肝病毒(SealHAV_NL/PV/21)。对来自同一地区的88只斑海豹和12只灰海豹的肝脏样本进行RT-PCR筛选,鉴定出9只斑海豹幼海豹肝病毒,VP1基因序列发生了微小变异。全基因组序列分析表明,SealHAV_NL/PV/21与新英格兰海豹肝病毒的核苷酸同源性为95.6%,但其5'-UTR长度增加了51 bp。系统发育分析表明,海豹肝病毒聚集在一个单系群中,与已在陆生哺乳动物中发现的其他肝病毒物种不同。对所有感染动物器官中海豹肝病毒RNA负荷的评估表明,肝脏的RNA拷贝数最高,每毫克组织中高达107个RNA拷贝。在肝脏的肝细胞中,通过原位杂交很容易检测到海豹肝病毒RNA,但与病理病变无关。采用基于hav的酶联免疫吸附试验(ELISA)对90份当代海豹血清进行血清学筛选,结果显示14份斑海豹和1份幼年灰海豹血清中存在肝病毒抗体。这些发现共同表明,海豹肝病毒在北海海豹中是地方性的,在幼小动物中引起静息性感染。
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