Cucumber mosaic virus (CMV) poses a considerable threat to a diverse array of crops in global agriculture. CMV impacts commercially important cut lilies by diminishing both yield and flower quality. We used RNA sequencing (RNA-seq) to investigate the changes in gene expression in the leaves and bulbs of four distinct cultivars of cut lily, ‘Cancun,’ ‘Brunello,’ ‘Connecticut King,’ and ‘Casa Blanca’ following CMV infection. Notably, CMV affected photosynthetic processes by significantly downregulating genes associated with photosynthesis. In addition, CMV infection was detrimental to chloroplast function and energy production. We observed differential expression of genes associated with both dominant and recessive resistance pathways that are crucial for preventing virus entry, replication, and systemic spread within the plant. Based on functional annotation and differential gene expression analysis, we identified the regulatory genes involved in triggering immune responses, modulating signal transduction, and specific host factors during CMV infection. To validate the RNA-seq findings, we selected four genes involved in resistance, virus multiplication, and virus spread and analyzed them using real-time quantitative reverse transcription PCR (qRT-PCR) with specific primers. The qRT-PCR results aligned closely with those from RNA-seq, showing consistent fold-change responses for the genes that were differentially expressed, indicating that the RNA-seq results were reliable. These results deepen our understanding of the complex genetics behind plant-virus interactions while also providing information for breeding programs that aim to develop CMV-resistant lily cultivars.

Because intact FMDV particles (146S) are often unstable in vitro, stabilizing foot-and-mouth disease virus (FMDV) antigens remains a key challenge in studying viral charateristics. Therefore, finding optimal condition to stabilize the FMDV is essential. In this study, we investigated formulations and potentials of several stabilizers such as appropriate buffer, excipients, and storage conditions to enhance the stability of 146S. Inactivated FMDV O-Jincheon (O-JC) was dissolved in various buffer formulations, and stored at 4℃ for two months to evaluate quantity of 146S at every 2-week interval. Among phosphate buffered saline (PBS), Tris buffered saline (TBS), HEPES buffered saline (HBS), and MOPS buffered saline (MBS), PBS showed more effective 146S stabilization that showed 1.3-1.6 fold higher 146S fraction than TBS, HBS, and MBS after storage for 2 weeks. However, constant dissociations of 146S were observed in all formulations at 8 weeks. Compared with other FMDVs, A22 Iraq and SAT-1, in PBS, O-JC proved to be the least stable in PBS. A variety of excipients including carbohydrate, sugar alcohol, cryo-protectant were tested for the capability in protecting O-JC from dissociation. By adding 4-8% sucrose, more than 60% of 146S fractions were maintained at 8 weeks, those were at least 1.8 fold higher than the PBS-only control. Addition of 1% β-cyclodextrin showed synergistic enhancement in O-JC stability. As the results of this study, it could be suggested that the PBS-based buffer together with 4-8% sucrose + 2% sorbitol or 2% sucrose + 2% sorbitol + 1% β-cyclodextrin could help the better stability of the O-JC in vaccine preparation.

The Japanese encephalitis virus (JEV) is a zoonotic pathogen that affects the nervous systems of humans, pigs, and horses. It has been classified into five genotypes (G1-G5) based on molecular analysis of the pre-membrane or envelope gene. In the Republic of Korea, the predominant JEV genotype has recently shifted from G3 to G1 and G5, highlighting the need for a rapid and accurate diagnostic method. In this study, we designed specific common and differential primer sets for JEV G1, G3, and G5 to detect the JEV gene. Four specific primer sets for JEV G1, G3, and G5 were used to selectively amplify the target gene. The detection limits of the common primer set for JEV G1, G3, and G5 were 100, 0.1, and 10 TCID50/reaction, respectively. The detection limits of the three differential primer sets were 1, 0.1, and 1 TCID50/reaction, respectively. No cross-reactivity was observed with non-JEV reference viruses. We successfully developed a multiplex reverse transcription polymerase chain reaction (RT-PCR) assay to distinguish the three JEV genotypes. Our multiplex RT-PCR assay is highly sensitive and specific, providing a reliable tool for confirming JEV infection in suspected samples. Additionally, our assay can be applied to suspected mosquito samples and commercial veterinary biological products.

Severe Fever with Thrombocytopenia Syndrome (SFTS) is a newly emerging tick-borne disease caused by the SFTS virus (SFTSV), which belongs to the phlebovirus in the Bunyaviridae family. SFTSV is enveloped with a tripartite ambisense RNA genome. The L segment encodes the viral RNA-dependent RNA polymerase, the M segment encodes the two glycoproteins, Gc and Gn, and the S segment encodes the nucleoprotein (NP) and the nonstructural protein (NSs). NP participates in ribonucleoprotein (RNP) packaging and commonly detected early after infection, suggesting that the N protein is possible to be used as a target antigen for early diagnosis of SFTSV infection. In this study, we analyzed a highly immunogenic multi-epitope using GnGc and NP genes from a consensus sequence of SFTSV strain isolated from infected patients in Korea. The selected genes are constructed to the expression vector plasmid pJHL65 and the recombinant plasmid vector was transformed into the Δasd Δlon ΔcpxR Salmonella Typhimurium attenuated strain JOL912 and the expression of these antigens was verified by immunoblotting assay. We observed the significant levels of systemic IgG and mucosal IgM responses against the JOL912-derived antigen in the immunized BALB/c mice. The level of CD3+CD4+, CD3+CD8+ T lymphocyte subpopulation and TNF-α were also highly regulated in splenic T cells re-stimulated in vitro with NP and Gn/Gc multi-epitope selected antigens. Therefore, immunized mice with NP and Gn/GC multi-epitope recombinant proteins of attenuated Salmonella delivery system elicited T cell-related immune response, inducing an effective immune response. In conclusion, the attenuated Salmonella expressing NP-GnGc multi-epitopes could be a novel vaccine candidate against the SFTS virus.

This study evaluated the virucidal efficacy of a chlorine dioxide (ClO2) gas-generating fumigation disinfectant consisting of sodium chlorate solution (25% sodium chlorate) and reaction solution against avian influenza virus (AIV). After AIV suspensions had been deposited on stainless steel carriers, the 9 dried carriers were exposed to the fumigant (sodium chlorate solution: 8.5, 17, 34, 50, and 100 mL) in a 25-m³ test room for 2, 3, and 4 h, respectively. Thereafter, all carriers were submerged in a neutralizing solution (20% fetal bovine serum) to scrape off the surviving viruses, and the respective suspensions were diluted. Each diluent was inoculated into the allantoic membrane of five 10-day-old embryos. After incubation for 5 days at 37℃, AIV viability in the collected allantoic fluids was examined, and the egg infectious dose 50 (EID50) was calculated. When the carrier was exposed to ClO2 gas generated from reacting 34 mL of the fumigant for 3 h, the AIV titer reduced by more than 104.0 EID50/carrier compared to the control, which was exposed to the fumigant without inoculation of AIV suspension. In addition, the control was non-toxic to the embryos.

바이러스는 생물 의약 산업에서 다양한 응용 분야를 가지고 있다. 그들은 살충제 생산, 백신 생산, 유전자 전달, 암 치료제 등에 사용된다. 바이러스의 하류 처리는 그들의 생물학적 및 의약적 응용을 위한 필수 단계이다. 다양한 과정 중에 서 바이러스의 정제는 매우 중요하다. 막 크로마토그래피는 이 과정에서 중요한 역할을 한다. 이온 교환 막 크로마토그래피는 주로 사용되는 방법이지만 크기 배제 및 불충분한 정제에 관한 다양한 제한을 가지고 있다. 또한, 이는 인플루엔자와 같은 빠 르게 변화하는 바이러스의 균주에 적용될 수 없다. 이 검토는 막 크로마토그래피의 다양한 개선된 방법 또는 대안을 검토한 다. 이는 정제, 바이러스 회수율 및 방법의 확장성에 초점을 맞추고 있다.