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Partial Sequence of a Novel Virus Isolated from Pelodiscus sinensis Hemorrhagic Disease
1Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Fish Health and Nutrition of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou, China.
Abstract:
Outbreaks of hemorrhagic syndrome-like disease with high mortality rates have frequently occurred in Pelodiscus sinensis farms. The purpose of this study was to investigate the pathogen through challenge infection assays and partial sequencing of the genome of the pathogen. A 453-bp amplicon was obtained by random PCR using the nucleic acid extracted from the tissue homogenate filtrate and showed 32% identity at the amino acid level with the replicase polyprotein of the porcine reproductive and respiratory syndrome virus by Blastx. Multiple alignments indicated the putative protein sequence has some similarities to the replicase polyprotein of Arteriviridae, and the phylogenetic tree showed it was closely related to equine arteritis virus. This sequence was found in the lung of the diseased P. sinensis by in situ hybridization. Dot blot hybridization and quantitative RT-PCR showed that the lung had the highest content of virus. The peak replication of P. sinensis hemorrhagic syndrome virus (TSHSV) in the lung occurred 4 days after infection. The ribonucleic nature of the viral genome was confirmed by RNase A or DNase I treatments. We named the virus TSHSV in this study as P. sinensis is also known as Trionyx sinensis. These results provide a fundamental basis for further understanding the biology and the molecular mechanisms of TSHSV.
Insights
A novel virus causing hemorrhagic syndrome in Chinese softshell turtles (Pelodiscus sinensis) was identified. This discovery provides crucial insights into turtle diseases and viral pathogens affecting aquaculture.
Area of Science:
- Virology
- Aquaculture Pathology
- Molecular Biology
Background:
- Frequent outbreaks of hemorrhagic syndrome-like disease with high mortality rates have been observed in Pelodiscus sinensis farms.
- The causative agent of these outbreaks remained unidentified, posing a significant threat to turtle aquaculture.
Purpose of the Study:
- To identify and characterize the pathogen responsible for hemorrhagic syndrome in Pelodiscus sinensis.
- To investigate the viral genome and its replication characteristics within infected turtles.
Main Methods:
- Challenge infection assays were performed on Pelodiscus sinensis.
- Partial viral genome sequencing using random PCR and Blastx analysis.
- In situ hybridization, dot blot hybridization, and quantitative RT-PCR were employed to detect and quantify the virus.
- RNase A and DNase I treatments were used to determine the nature of the viral genome.
Main Results:
- A 453-bp amplicon was obtained, showing partial homology to Arteriviridae replicase polyproteins, particularly equine arteritis virus.
- The virus, named Pelodiscus sinensis hemorrhagic syndrome virus (TSHSV), was detected primarily in the lungs of infected turtles.
- Peak viral replication in the lungs occurred at 4 days post-infection, and the viral genome was confirmed to be ribonucleic acid (RNA).
- Quantitative RT-PCR indicated the highest viral content in the lung tissue.
Conclusions:
- A novel RNA virus, TSHSV, has been identified as the causative agent of hemorrhagic syndrome in Pelodiscus sinensis.
- The study provides foundational data for understanding TSHSV biology and molecular mechanisms, crucial for disease management in aquaculture.
