• 한국어병학회지
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• 제33권2호
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• pp.103-109
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• 2020

In order to use nervous necrosis virus (NNV) virus-like particles (VLPs) as a delivery tool for heterologous antigens or plasmids, we attempted to produce red-spotted grouper nervous necrosis virus (RGNNV) VLPs displaying a partial region of viral hemorrhagic septicemia virus (VHSV) glycoprotein at the surface and VLPs that are harboring DNA vaccine plasmids within the VLP. A peptide encoding 105 amino acids of VHSV glycoprotein was genetically inserted in the loop region of NNV capsid gene, and VLPs expressing the partial part of VHSV glycoprotein were successfully produced. However, in the transmission electron microscope analysis, the shape and size of the partial VHSV glycoprotein-expressing NNV VLPs were irregular and variable, respectively, indicating that the normal assembly of capsid proteins was inhibited by the relatively long foreign peptide (105 aa) on the loop region. To encapsulate by simultaneous transformation with both NNV capsid gene expressing plasmids and DNA vaccine plasmids (having an eGFP expressing cassette under the CMV promoter), NNV VLPs containing plasmids were produced. The encapsulation of plasmids in the NNV VLPs was demonstrated by PCR and cells exposed to the VLPs encapsulating DNA vaccine plasmids showed fluorescence. These results suggest that the encapsulation of plasmids in NNV VLPs can be done with a simple one-step process, excluding the process of disassembly-reassembly of VLPs, and NNV VLPs can be used as a delivery tool for DNA vaccine vectors.

• 대한수의학회지
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• 제46권1호
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• pp.21-25
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• 2006

Iridovirus is an icosahedral cytoplasmic double-stranded DNA virus with a genome size of 170-200kb. Outbreaks of fish iridovirus infection are characterized by their wide geographic distribution and broad host spectrum, especially in water temperatures of $25-27^{\circ}C$ Recently, the causative agent of high mortalities in flounder (Paralichthys olivaceus) was identified as fish iridovirus in Korea. Iridoviral infection repeatedly occurs in the same area for long periods, suggesting the possibility of viral infection in nursery. To examine this, the existence of iridovirus in juvenile flounders was detected by PCR using virus-specific primers. Antibodies induced against this virus were also examined using ELISA. As a result, juvenile fish in nursery were found to be previously infected with iridovirus, suggesting that proper prevention systems are required.

• Journal of Microbiology and Biotechnology
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• 제33권11호
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• pp.1412-1419
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• 2023

Retroviral integration into ancient vertebrate genomes left traces that can shed light on the early history of viruses. In this study, we explored the early evolution of retroviruses by isolating nine Spuma endogenous retroviruses (ERVs) and one Epsilon ERV from the genomes of Agnatha and Chondrichthyes. Phylogenetic analysis of protein sequences revealed a striking pattern of co-evolution between jawless fish ERV and their host, while shark ERV underwent ancient cross-class viral transmission with jawless fish, ray-finned fish, and amphibians. Nucleotide sequence analysis showed that jawless fish ERV emerged in the Palaeozoic period, relatively later than ray-finned fish ERV. Moreover, codon analysis suggested that the jawless fish ERV employed an infection strategy that mimics the host codon. The genealogical diversity of ERVs in the jawless fish genome highlights the importance of studying different viral species. Overall, our findings provide valuable insights into the evolution of retroviruses and their interactions with their hosts.

• 한국어병학회지
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• 제13권2호
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• pp.121-127
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• 2000

1998년 8~10월에 남해안 일원 해상 가두리 양식장에서 사육중인 돌돔, Oplegnathus fasciatus이 60%이상 폐사하였다. 병어의 체색은 검어지거나 퇴색되며 아가미는 빈혈증상이 심하고 비장은 특이적으로 비대되어 있었고, 비장, 신장, 심장 및 간장 조직에서 퓰겐양성과 호염기성의 비대세포가 관찰되었다. 병어의 비장조직 마쇄액을 GF세포에 접종배양한 결과, 세포가 구형화 되면서 커지는 세포변성효과가 나타나 감염된 GF 세포로부터 바이러스가 분리되었다. 분리된 바이러스 배양액($10^4TCID_{50}$/0.1 ml)을 건강한 돌돔 치어에 복강주사하면 자연감염어와 유사한 증상을 나타내면서 폐사가 일어났고, 또한 병어의 비장조직을 전자현미경으로 관찰한 결과, 조직내 비대세포의 세포질에서 직경이 120~130 nm이며 외막이 있는 정 20면체의 바이러스 입자가 관찰되었다. 그리고 genebank로부터 얻은 참돔 이리도바이러스(RSIV)의 ATPase gene의 DNA염기서열을 주형으로 제작된 primer를 사용하여 PCR을 수행한 결과, 500 bp의 PCR 생성물이 병어나 인위감염된 GF세포에서 얻어졌는데, 이들 PCR 생성물은 RSIV의 ATPase cDNA gene과 95%의 상동성을 나타내었다. 따라서 양식돌돔에 대량 폐사를 일으킨 원인 바이러스는 참돔 이리도바이러스(RSIV)와 유사한 이리도바이러스로 밝혀졌다.

• 한국수산과학회지
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• 제51권3호
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• pp.328-331
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• 2018

We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against nervous necrosis virus (NNV, RGNNV genotype). We established six hybridoma clones secreting MAbs against NNV antigen: 2B1, 2B11, 2C12, 13C1-1, 13C1-2 and 14D11. All six MAbs belonged to the IgG2a isotype with a kappa light chain and their reactivity recognized against the 41 kDa coat protein of NNV by Western blot analysis. The affinity constants of the six MAbs were measured by enzyme-linked immunosorbent assay (ELISA). All six MAbs reacted with two NNV isolates (SgNag05 and Gemunodo06), while no reactivity was observed with five know fish viruses, namely marine birnavirus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, hirame rhabdovirus, and infectious hematopoietic necrosis virus. Moreover, high ELISA optical density (OD) values (0.87-1.42) were observed in the brain tissues of NNV-infected sevenband grouper, while low OD values (less than 0.12) were recorded in the brain tissues of uninfected fish. These results suggest that these six MAbs are highly competent and useful for the detection of NNV with the RGNNV genotype.

• 한국어병학회지
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• 제11권2호
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• pp.89-96
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• 1998

해양버나바이러스(MABV)는 여러 종의 해양생물에 감염되며 숙주역이 넓다. 다양한 종의 숙주에 감염되었을 때의 MABV의 감염 특성을 규명하기 위하여, 주화세포 내에서 바이러스를 10대 계대 배양하여 in vitro로 연구했다. CHSE-214, RTG-2와 RSBK-2세포에서는 전형적인 CPE를 보이며 많은 양의 바이러스가 생산되었고, 높은 바이러스 단백질의 발현도 관찰되었다. 이에 반하여, EPC, FHM과 BF-2세포에서는 형광항체법에 의하여 바이러스단백질은 검출되었으나 CPE는 나타나지 않았다. EPC와 FHM 세포에서는 계대를 할수록 바이러스의 역가가 높아져, 바이러스의 숙주세포에의 적응이 일어난 것으로 보인다. 플라크의 크기는 CHSE-214, RTG-2와 RSBK-2세포에서 계대한 것이 다른 세포에서 계대한 것보다 커, 숙주세포의 종류에 따른 변이가 바이러스에 일어난 것으로 추측되었다. 게놈분절 A에 존재하는 VP2/NS 경계영역의 염기배열에서는 195번째의 염기에 특이적인 변이가 보였다. 숙주세포의 종류에 따라 다른 MABV의 감염특성은 자연계에서 다양한 숙주종에서 일어나는 in vivo에서의 감염특성을 반영하는 것으로 생각된다.

• 한국어병학회지
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• 제13권1호
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• pp.7-13
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• 2000

1993년부터 한국 서해안의 새우 양식장에서는 양식새우의 대량폐사가 일어났다. 외부증상은 두흉갑과 표피에 흰 반점이 나타났고 어류주화세포에는 배양되지 않았다. 열($50^{\circ}C$) 및 강산(pH 3)에는 쉽게 실활되었으나 강알카리(pH 11)에는 내성이 강했다. 바이러스의 입자 형태는 Rod Shaped한 형태를 보였다. 바이러스 단백질의 분석결과는 Hypodermal Hematopoietic Necrosis Baculovirus(HHNBV)와 유사했고 바이러스 핵산분석 결과는 약 114kb로 Penaeid Acute Viremia(PAV)와 유사했다.

• 한국어병학회지
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• 제17권1호
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• pp.1-10
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• 2004

해수 환경 중의 VHSV 분포 조사를 위하여, 2003년 2월에서 5월까지 동해 및 남해안 인근 해역에 서식하는 자연산 해수 어류를 채집하였다. 바이러스 분리용 시료는 9종의 해수 어류로서 42개의 시료를 분석하였다. 각 시료의 조직여과액을 Epithelioma papulosum cyprini (EPC) cells에 접종한 결과, 고등어 Scomber japonicus와 숭어 Mugil cephalus에서 바이러스가 분리되었으며 이들 바이러스는 RT-PCR법을 이용하여 VHSV로 동정할 수 있었다. 고등어에서 VHSV가 검출되었다는 기보고는 있지만, 숭어에 대해서는 본 연구에서 처음으로 VHSV를 분리하였다. 자연산 어류에서 유래된 세 개의 VHSV isolates에 대한 glycoprotein gene을 분석한 결과, 이들 바이러스는 양식 넙치에서 질병을 유발하는 VHSV isolates와 유사하며 모두 genogroup I (American type)에 속하는 것으로 밝혀졌다.

• 한국수산과학회지
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• 제54권5호
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• pp.596-616
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• 2021

A global increase in fish consumption has led to a rapid expansion of aquaculture production, which has been linked to enhancing the spread of infectious diseases. Viral diseases can cause high mortality in many cultured fish species, posing a serious threat to the aquaculture industry. Infectious hematopoietic necrosis virus (IHNV) is one of the primary threats to aquacultured salmonid species, causing huge economic losses. Since the first report in cultured sockeye salmon Oncorhynchus nerka during the 1950s in North America, IHNV has spread to other regions, including Europe, Asia, South America, and Africa by transportation of infected fish and eggs, causing disease and increasing mortality in a wide variety of salmonid species. Here, we review existing information relevant to IHNV: its phylogenetic characteristics, origin, infection history, virulence determinants, susceptible hosts, vectors, and vaccine development. This review also addresses a possible cross-species transmission of IHNV to a new host, olive flounder Paralichthys olivaceus, a cultured fish of economic importance in East Asian countries.

• 한국어병학회지
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• 제37권1호
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• pp.9-15
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• 2024

An enzyme-linked immunosorbent assay (ELISA) with two Novirhabdovirus antigens (viral hemorrhagic septicemia virus, VHSV and infectious hematopoietic necrosis virus, IHNV) was used to detect specific antibodies against VHSV from olive flounder (Paralichthys olivaceus) sera. In ELISA plates with VHSV culture supernatants (VHSV-Ag plate), optical density (OD) values for sera from olive flounder with VHS history (VHS sera) ranged from 0.64±0.36, and those of sera from fish without VHS history (non-VHS sera) ranged from 0.26±0.26. In IHNV-Ag plate, the OD values (0.43±0.28) for VHS sera were quite low compared to those in VHSV-Ag plates, while the OD values for non-VHS sera were almost similar. When the OD values for each serum were calculated by subtracting the OD values in the IHNV-Ag plate from those in the VHSV-Ag plate, the corrected OD values were significantly different between VHS sera and non-VHS sera. The results were completely in line with fish histories of VHS epizootics. It was considered that the corrected OD values may represent the true values recognized by VHSV-specific antibodies.