Hand, foot, and mouth disease (HFMD) is a highly contagious disease with no specific treatment. Since it is common in immunocompromised children under the age of 5, there is a need to develop a safe vaccine. Virus-like particles (VLPs) are similar structures to viruses with the lack of genetic material which makes them impossible to replicate and infect, and therefore have a high level of biological safety and are considered to have high value as vaccines. In this study, the insect virus expression system that is widely used for vaccine and drug production due to its high post-translational modification efficiency, was used to produce VLPs for Coxsackievirus type A6 and A10, which are recently reported to be the main causes of HFMD. For this purpose, the selection of promoters that can control the timing and intensity of expression of 3CD protein, which is essential for VLPs assembly but has been reported to be cytotoxic, was conducted to construct an optimal expression form for HFMD-VLP.

Foot-and-mouth disease (FMD), which affects cloven-hoofed animals, is economically important because of its highly contagious nature. FMD virus (FMDV), the causative agent of FMD, involves seven serotypes (O, A, Asia1, C, and SAT 1-3). Serotype Asia1 is unique to the Asian territory and is subdivided into nine genetic groups (G-I-IX) based on nucleotide variations in the VP1 sequence. Asia1 Shamir, the most representative Asia1 vaccine, is not highly protective against the Asia1/MOG/05 (G-V) lineage found in North Korea in 2007. Therefore, we investigated whether a chimeric virus strain (Asia1/MOG/Shamir), in which the VP4, VP2, and VP3 sequences of Asia1/MOG/05 were combined with the VP1 sequence of Asia1 Shamir, can simultaneously protect against both viruses. We determined the optimal viral growth conditions for the commercial utilization of this chimeric virus strain. Of the three types of cell culture media, the Cellvento medium resulted in the highest amount of antigen in the samples. The chimeric strain was proliferated in a small bioreactor to produce a test vaccine, and its immunogenicity was evaluated in pigs. The virus neutralization (VN) titer against the Asia1 Shamir virus was > 1/100 after the second immunization with the chimeric vaccine in pigs. In addition, a single dose of the test vaccine resulted in a VN titer of > 1/100 against the Asia1/MOG/05 strain. Taken together, our chimeric vaccine strain provided sufficient protection against the Asia1/MOG/05 and Asia1 Shamir viruses, suggesting its potential as a novel vaccine for both these strains.

The Republic of Korea has implemented an obligatory vaccination on major livestock breeds to prevent and eradicate nationwide foot and mouth disease (FMD) since the end of 2010. The government has executed massive serological survey to check the immune level of various herds after vaccination, and seropositive rates against FMD virus (FMDV) structural proteins can be measured to assess FMD immunity level. The purpose of this study is to investigate the FMDV serological level of every cattle breeding farm in the country and to determine whether there is a significant difference between groups classified by time, age, and management authority. A total of 5,781 serum samples was collected in 18 cattle breeding farms from 2020 to 2021, and the seropositive rates were measured using PrioCHECK FMDV Type O ELISA kit. Firstly, the cattle breeding farms were classified by which they are managed: the central government, the local government, and the private agency. Every management authority had a seropositive rate of 99.5% or higher. Secondly, the samples were divided into 6 to 12 months old, 12 to 24 months old, and 24 months or more. The 6 to 12 months old group in 2020 showed a significantly low seropositive rate of 98.1%, but it was improved by implementing the enhanced vaccination policy from 2021 to 100%. In summary, there are considerably high seropositive rates including all groups with time, age, and by which they are managed, which means the FMD vaccination in cattle breeding farms is well-managed.

Foot-and-mouth disease (FMD) is a highly contagious vesicular disease that affects cloven-hoofed animals, causing substantial economic losses to the livestock industry. The causative FMD virus (FMDV) comprises four structural proteins (VP1, VP2, VP3, and VP4) and several non-structural proteins. Among the capsid proteins, VP4 is the most conserved, making it an attractive target as a diagnostic and vaccine antigen, regardless of FMDV serotype. In this study, we attempted to express the VP4 protein N-terminally fused to a glutathione S-transferase (GST) tag in Escherichia coli. Whereas VP0 and VP2 proteins were expressed in the soluble fraction, we failed to detect VP4, even in the insoluble fraction. To investigate the effect of VP4 C-terminal amino acid residues on protein expression, we constructed three VP4 mutants fused to GST, among which the mutant in which the C-terminal 15 amino acid residues had been deleted showed the highest level of protein expression. Furthermore, protein expression was observed even in the mutant in which three amino acid residues (DKK) had been fused to the C terminus. However, unlike the other two mutants, the wild-type VP4 mutant was poorly expressed, thereby indicating that the C-terminal amino acid residues could play a pivotal role in determining expression of the VP4 protein in E. coli.

Vaccinations, surveillance, quarantine, and disposal of the infected poultry are the common strategies for prevention and control of the highly infectious poultry diseases; however, many pathogens still persist and are potential causes threatening the nationwide spread of poultry diseases. A strict biosecurity strategy including disinfection is the key to control the spread of avian pathogens, such as the Newcastle disease virus (NDV). It is important to select and use the disinfectants whose efficacy and the effective concentrations against the specific pathogens are known. Therefore, in the present study, we evaluated the virucidal efficacy of five active substances of commercial disinfectants, namely potassium peroxymonosulfate (PPMS), sodium dichloroisocyanurate (NaDCC), glutaraldehyde (GLT), benzalkonium chloride (BZK), and didecyldimethylammonium chloride (DDAC), that are used against NDV. Further, we validated the efficacy of eight pre-approved disinfectants. The minimal virucidal concentrations of the active substances against NDV were as follows: PPMS, 2.50 g/L; NaDCC, 2.00 g/L; GLT, 0.40 g/L; BZK, 2.00 g/L, and DDAC, 1.00 g/L. Furthermore, all the eight disinfectants were found to be effective against NDV at the recommended concentrations, thereby confirming that the active substances are functional against NDV. This is the first study reporting the virucidal activity of the active substances of commercial disinfectants against NDV, in accordance with the Animal and Plant Quarantine Agency guidelines, in South Korea. The results of the virucidal efficacy testing of chemical disinfectants from this study will help poultry industries implement improved strategies for controlling infection.

This study is a descriptive correlation study to explore the intention of nursing for patients with COVID-19 among nurses in a tertiary hospital in Seoul, and to identify factors influencing the nursing intention. Data were collected from 239 nurses with more than 6 months of working experience from December 1, 2020 to December 31, 2020. Data were analyzed by t-test, ANOVA, Scheffé test, Pearson's correlation coefficient, and Hierarchical multiple regression using the SPSS WIN 24.0 program. According to a result of this study, the score of participant's nursing intention for patients with COVID-19 was an average of 4.48 points out of 7 points. Factors influencing the nursing intention were control beliefs, position, positive behavior beliefs, negative behavior beliefs, and normative beliefs: Among the factors influencing nursing intentions, control beliefs have been shown to have the greatest influence. These variables accounted for 65.2% of the variance in the nursing intention. Therefore, to increase the nursing intention for patients with COVID-19, it is necessary to educate and train nurses to strengthen their confidence in caring the patients. In addition, repeated studies were suggested with extended sample in other settings and to find other factors influencing the nursing intention.

In the case of foot-and-mouth disease (FMD), there is a great deal of impact on the national economy due to the disposal of diseases, the cost of disease control such as vaccination, reduction of productivity, and restriction of international trade of livestock products. Therefore, appropriate diagnostic methods for sensitive, accurate and rapid identification of virus serotypes are continuously required in terms of early prevention of FMD. This study was conducted to confirm the feasibility of immuno-PCR diagnostic method for the more sensitive detection of Korean FMD virus (FMDV). We synthesized a partial FMD type A viral gene. Protein antigen, monoclonal and polyclonal antibodies of FMDV were cloned, expressed and purified and then magnetic particles were attached to polyclonal antibodies and and oligomers to monoclonal antibodies for the immnuno-PCR. We confirmed the antigen-antibody and oligomer reaction using ELISA, Western blot, and real-time PCR. These results show that Korean FMDV can be detected by using difference of Ct values between positive group and negative group using immuno-PCR.. The results of this study also suggest that this technique will be the basis of the diagnosis method to detect Korean FMDV more sensitively in the future.

Sacbrood virus (SBV) caused significant colony collapse in Korean Apis cerana. Considering that hygienic behavior in honey bees confers colony-level resistance against brood diseases, we utilized this trait for selecting A. cerana colonies. In addition, the brood survival rate was evaluated after colonies were SBV-inoculated. Over four selective generations, dead brood removal and brood survivorship in selected colonies were higher than those in the unselected colonies (P < 0.01, 99.3 vs. 89.9% for removal of pin-killed pupae; P < 0.01, 99.0 vs. 63.9% for removal of SBV-killed larvae; and P < 0.01, 70.0 vs. 9.2% for brood survivorship). Following SBV-inoculation, selected colonies showed an increase in the number of surviving pupae and adults, whereas unselected colonies collapsed mostly. Our results confirm the feasibility of selecting SBV-resistant A. cerana.

미세방울 디지털 PCR(Droplet digital PCR, ddPCR) 법은 정량 PCR 법과 같이 형광물질을 사용하여 정량분석이 가능하다는 점은 유사하지만, PCR 반응액을 만든 후 이 반응액을 바로 PCR에 사용하지 않고 수 만 개의 동일한 크기의 미세방울(droplet)로 구획(partitioning)한 후에 PCR 반응을 수행하는 점이 기존의 PCR 법과 가장 큰 차이라고 볼 수 있다. 이렇게 구획된 나노리터 크기의 미세방울 중에서 형광신호가 검출되는 미세방울과 검출되지 않은 미세방울의 수를 계수하여 프아송 분포(Poisson distribution) 계산에 적용하면 표준검량선 없이 목적 유전자의 절대정량 분석이 가능하다. 곤충이 매개하는 질병 바이러스의 경우 소량의 바이러스 감염 여부를 확인하기 위해서 다수의 곤충 유전자를 확인해야 하는데, 이와 같이 미세방울 형태의 ddPCR을 이용하면 기존 Real-time PCR 법에 비해 극소량의 목적 유전자를 높은 민감도로 검출할 수 있으며, PCR 저해요소(inhibitor)에도 큰 영향을 받지 않는다는 장점이 있다. 또한 미세방울 방식의 디지털 PCR을 이용하면 다중 PCR 분석이 가능하여 1개의 시료에서 다양한 질병매개 바이러스를 검출할 수 있다.

A cell line of bovine origin was immortalized to isolate foot-and-mouth disease virus (FMDV). The immortalization was performed by infection of bovine primary epithelial cells with a recombinant retrovirus that overexpressed the human telomerase (hTERT), after primary culture of fetal bovine kidney tissue and removal of fibroblasts. After cloning the immor- talized cell line into single cells, the cloned cell lines were named JNUBK-1, JNUBK-2, JNUBK-3 and JNUBK-4, according to their characteristics. To confirm the epithelial phenotype of the cell lines JNUBK-3 and JNUBK-4, which showed stable proliferation capability over 35 generations after immortalization, the expression of cytokeratin and fibronectin was measured. Finally, the FMDV titer in the JNUBK-3 and JNUBK-4 cell lines was measured and was 800∼2,000 times higher than that of the currently used cell line IRBS-2. In conclusion, more sensitive isolation and production of FMDV became possible through the use of the immortalized JNUBK-3 and JNUBK-4 cell lines.

구제역 바이러스는 소나 돼지 등의 우제류 가축에 경구 또는 흡입 노출 등의 다양한 노출 경로를 통해 심각한 감염성을 유발하는 생물학적 유해인자로 국내 축산업 운영에 있어 막대한 경제적 손실을 초래하는 병원균들 중 하나다. 구제역 발병에 대한 사전적 예방 관리를 도모할 수 있는 조치 방안 중 하나는 공기를 통해 전파되는 구제역 바이러스 확산에 대해 수학적 모델 적용을 통해 예측된 분석 결과에 따라 설정될 수 있다. 대기 확산 모델들은 일반적으로 구제역 바이러스의 주요 전파 경로인 경구 및 접촉 감염의 노출 시나리오를 예측할 수는 없으나, 상대적으로 예측도가 높은 대기 확산 모델의 경우 공기 전파에 대한 구제역 바이러스의 위해성 관리 방안을 결정하는 데 주요 역할을 담당할 수 있는 유용가능한 수단으로 여러 나라에서 활용되어 왔다. 구제역 바이러스 전파를 억제하기 위한 엄격한 관리 방안 중 다량의 가축 살처분 방법은 축산 농가들에게 심각한 경제적 손실을 초래하여 심각한 경우 새로운 자립의 여건마저도 상실시킬 수도 있다. 반면 낮은 수준의 대응 방안은 향후 구제역 발생에 따른 추가적 피해를 근원적으로 억제할 수 없는 역학적 한계에 봉착할 수 있다. 본 연구는 구제역 바이러스의 기본 특성 및 발생 모델 적용에 따른 공기 중 전파의 감염 위험성 평가 관련 선행 문헌들을 고찰하였다. 또한 1970년대 이후 구제역 바이러스의 공기 전파 경로를 예측하기 위해 여러 나라에서 실제 활용된 다양한 대기 확산 모델들의 적용 사례 및 장/단점을 비교 분석하였다.

Avian influenza (AI) and Newcastle disease (ND) distress a variety of avian species, especially domestic poultry. Severe syndromes are caused by highly virulent specific virus strains termed highly pathogenic AI and velogenic ND viruses, which are potential agrobioterrorism agents. This outbreak emphasizes the need for continuing cooperation between public health and veterinary medical communities in controlling AI and ND when it has a zoonotic potential. Up to date, the stamping out and burying system were applied for controlling methods against these highly infectious diseases in the ordinary way, however these methods had many environmental problems, including leachat and effluvium. Thus, we assessed that sterilization effect of AI and ND virus dependent on several treatment conditions, such as autoclaving time and cutting types of chicken. As a result, we found that the cutting type of chicken meat revealed a reduced HA titer (20) against both of AI and ND virus after 10 min of autoclaving, while whole chicken showed same titer after 30 and 60 min. Therefore, we propose that the conditions of treatment on infected chicken should be developed for convenient, affordable, and effective prevention methods against for AI and ND.

The lung and lymph node samples were collected from 786 pig farms associated with wasting and respiratory syndrome during 2005～2009. All samples were tested for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) and the differentiation of its genotype using reverse transcriptase-polymerase chain reaction (RT-PCR). 643 farms (81.8%) of the pig farms examined were positive for PRRSV, of which 57.2% accounted for PRRSV type 1 and 70.2% accounted for PRRSV type 2. Furthermore, 37.5% of the farms positive for PRRSV, showed the coexistence of two genotypes. The results indicate that the PRRSV infections of single genotype or two genotypes are very common in Korean pig farms.

In many cases over the years, we failed to forecast accurately outbreak of Rice stripe virus (RSV) disease using population density of overwintering small brown planthopper (SBPH), Laodelphax striatellus. What is the source of error in this viral disease assessment? For answering the question, spatial distribution of RSV in paddy fields was investigated in relation to the yield of rice grain, and the population density of overwintering SBPH in ridge of paddy fields. Total 14 paddy fields in 5 regions were surveyed in June, 2008. Disease assessment was carried out with each of 30 rice plants in the one of the borders, 5th, 10th line from ridge, and 90 rice plants in diagonal line for conventional method. The ears of rices from 18 plants in the same surveyed line were collected in the late of August for the weights of 100 rice grains. The infection rate of RSV was decreased by the distance from ridge; on the contrary, the increase tendency was shown in the yield of rice grains in the middle of paddy field. It is suggested that the border effect of viral distribution between ridge and paddy field is caused by the spatial distribution of the vector insect. Thus, in this case, the narrow border effect could represent that the SBPH migrated short distance from the ridge to paddy field. The conventional method, observing the disease in middle area of diagonal line, showed no correlation to the population density of overwintering vector generation in ridge. It might cause underestimation of the regional disease rate. The counting of RSV-diseased plant in border line only overestimated it also. However, the RSV infection rate in the near border had highly positive linear relationship (R2=0.91) to the regional average values by regression analysis. Therefore, the near border sampling method can improve the RSV disease risk assessment.

Aujeszky’s disease (AD), also called pseudorabies, is an infectious viral disease, caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. AD affects many countries and regions in the world, causing important economic losses, mainly due to international trade restrictions. In this study, to determine the characteristics of the Aujeszky’s disease virus (ADV), NYJ strain, which was isolated from the serum of an infected pig in 1987, we investigated the nucleotide sequence and expression of the glycoproteins gB, gC, and gD using the bBpGOZA system. We found that the glycoproteins gB, gC, and gD of NYJ consisted of 2751 bp, 1443 bp, and 1203 bp, respectively. Comparison of the NYJ with the other strains revealed nucleotide sequence identity ranging from 91.tito 99.0%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The NYJ strain was formed a distinct branch with high bootstrap support. The expression of glycoprotein gD in insect cells was characterized by SDS-PAGE and Western blotting with an anti-ADV polyclonal antibody. Glycoprotein gD of approximately 45 kDa was detected. The results of this study have implications for both the taxonomy of ADV and vaccine development.