• 미생물학회지
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• 제54권3호
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• pp.254-265
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• 2018

최근 양계업에 막대한 피해를 끼치는 조류독감은 한국에서 수천억원의 거대한 경제적 손실을 초래하였다. 병원균의 전염경로를 파악할 수 있다면 막대한 손해를 끼치는 생물학적 피해의 확산을 막고 일부 지역으로 제한하는데 큰 도움이 될 것이다. 병원균 DNA 서열의 계통학적인 분석을 통하여 감염된 숙주들을 방향성이 있는 연결선으로 연관짓는 전염 계통수를 얻을 수 있다. 지난 10여년간 유전적 데이터뿐만 아니라 역학 데이터를 이용한 전염 계통수 추론의 방법론적 발전이 이루어졌다. 이에, 본 연구에서는 전염 계통수 추론 방법을 이용하여 지난 2014년 한국에 발병한 고병원성 조류독감 H5N8에서 유래한 DNA 서열을 재분석하였다. 당시, H5N8 바이러스는 전라북도에서 시작하여 지역적으로 접해있는 4개의 지역으로 확산되어 나갔던 것으로 알려져 있다. 전염 계통수를 추론하는 베이지언 통계 방법인 Markov chain Monte Carlo를 반복적으로 시행하고 이를 종합하여 철새 외래종과 국내종 조류 숙주들의 전염 계통수를 추정하였다. 비록 연결선의 불확실성은 높았으나 추정된 전염 계통수를 통하여 당시 H5N8 바이러스는 전라북도에서 시작하고 충청남도를 거쳐 경기도로 퍼져나간 것을 확인할 수 있었다. 사육하는 오리와 같은 국내종 조류는 전염 계통수의 말단 노드에 위치하는 것으로 추정되었다. 이러한 결과를 통하여 야생 철새종이 2014년 한국의 H5N8 조류독감의 감염 매개자로 주된 역할을 하였다는 것을 재확인하였다.

• Animal Bioscience
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• 제35권7호
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• pp.964-974
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• 2022

Objective: The highly pathogenic avian influenza virus (HPAIV) is a threat to the poultry industry and economy and remains a potential source of pandemic infection in humans. Antiviral genes are considered a potential factor for studies on HPAIV resistance. Therefore, in this study, we investigated gene expression related to the mitogen-activated protein kinase (MAPK) signaling pathway by comparing non-infected, HPAI-infected resistant, and susceptible Ri chicken lines. Methods: Resistant (Mx/A; BF2/B21) and susceptible Ri chickens (Mx/G; BF2/B13) were selected by genotyping the Mx and BF2 genes. Then, the tracheal tissues of non-infected and HPAIV H5N1 infected chickens were collected for RNA sequencing. Results: A gene set overlapping test between the analyzed differentially expressed genes (DEGs) and functionally categorized genes was performed, including biological processes of the gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. A total of 1,794 DEGs were observed between control and H5N1-infected resistant Ri chickens, 432 DEGs between control and infected susceptible Ri chickens, and 1,202 DEGs between infected susceptible and infected resistant Ri chickens. The expression levels of MAPK signaling pathway-related genes (including MyD88, NF-κB, AP-1, c-fos, Jun, JunD, MAX, c-Myc), cytokines (IL-1β, IL-6, IL-8), type I interferons (IFN-α, IFN-β), and IFN-stimulated genes (Mx1, CCL19, OASL, and PRK) were higher in H5N1-infected than in non-infected resistant Ri chickens. MyD88, Jun, JunD, MAX, cytokines, chemokines, IFNs, and IFN-stimulated expressed genes were higher in resistant-infected than in susceptible-infected Ri chickens. Conclusion: Resistant Ri chickens showed higher antiviral activity compared to susceptible Ri chickens, and H5N1-infected resistant Ri chickens had immune responses and antiviral activity (cytokines, chemokines, interferons, and IFN-stimulated genes), which may have been induced through the MAPK signaling pathway in response to H5N1 infection.

• Animal Bioscience
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• 제36권4호
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• pp.570-583
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• 2023

Objective: Fibroblast growth factors (FGFs) play critical roles in embryo development, and immune responses to infectious diseases. In this study, to investigate the roles of FGFs, we performed genome-wide identification, expression, and functional analyses of FGF family members in chickens. Methods: Chicken FGFs genes were identified and analyzed by using bioinformatics approach. Expression profiles and Hierarchical cluster analysis of the FGFs genes in different chicken tissues were obtained from the genome-wide RNA-seq. Results: A total of 20 FGF genes were identified in the chicken genome, which were classified into seven distinct groups (A-F) in the phylogenetic tree. Gene structure analysis revealed that members of the same clade had the same or similar exon-intron structure. Chromosome mapping suggested that FGF genes were widely dispersed across the chicken genome and were located on chromosomes 1, 4-6, 9-10, 13, 15, 28, and Z. In addition, the interactions among FGF proteins and between FGFs and mitogen-activated protein kinase (MAPK) proteins are limited, indicating that the remaining functions of FGF proteins should be further investigated in chickens. Kyoto encyclopedia of genes and genomes pathway analysis showed that FGF gene interacts with MAPK genes and are involved in stimulating signaling pathway and regulating immune responses. Furthermore, this study identified 15 differentially expressed genes (DEG) in 21 different growth stages during early chicken embryo development. RNA-sequencing data identified the DEG of FGFs on 1- and 3-days post infection in two indigenous Ri chicken lines infected with the highly pathogenic avian influenza virus H5N1 (HPAIV). Finally, all the genes examined through quantitative real-time polymerase chain reaction and RNA-Seq analyses showed similar responses to HPAIV infection in indigenous Ri chicken lines (R² = 0.92-0.95, p<0.01). Conclusion: This study provides significant insights into the potential functions of FGFs in chickens, including the regulation of MAPK signaling pathways and the immune response of chickens to HPAIV infections.

• Journal of Veterinary Science
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• 제24권1호
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• pp.13.1-13.11
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• 2023

Background: Highly pathogenic avian influenza viruses (HPAIVs) is an extremely contagious and high mortality rates in chickens resulting in substantial economic impact on the poultry sector. Therefore, it is necessary to elucidate the pathogenic mechanism of HPAIV for infection control. Objective: Gene set enrichment analysis (GSEA) can effectively avoid the limitations of subjective screening for differential gene expression. Therefore, we performed GSEA to compare HPAI-infected resistant and susceptible Ri chicken lines. Methods: The Ri chickens Mx(A)/BF2(B21) were chosen as resistant, and the chickens Mx(G)/BF2(B13) were selected as susceptible by genotyping the Mx and BF2 genes. The tracheal tissues of HPAIV H5N1 infected chickens were collected for RNA sequencing followed by GSEA analysis to define gene subsets to elucidate the sequencing results. Results: We identified four differentially expressed pathways, which were immune-related pathways with a total of 78 genes. The expression levels of cytokines (IL-1β, IL-6, IL-12), chemokines (CCL4 and CCL5), type interferons and their receptors (IFN-β, IFNAR1, IFNAR2, and IFNGR1), Jak-STAT signaling pathway genes (STAT1, STAT2, and JAK1), MHC class I and II and their co-stimulatory molecules (CD80, CD86, CD40, DMB2, BLB2, and B2M), and interferon stimulated genes (EIF2AK2 and EIF2AK1) in resistant chickens were higher than those in susceptible chickens. Conclusions: Resistant Ri chickens exhibit a stronger antiviral response to HPAIV H5N1 compared with susceptible chickens. Our findings provide insights into the immune responses of genetically disparate chickens against HPAIV.

• 한국농공학회논문집
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• 제54권1호
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• pp.63-72
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• 2012

In livestock industry, damage caused by the epidemic diseases such as Foot-and-Mouth Disease (FMD), Highly-Pathogenic-Avian-Influenza (HPAI) and Porcine-Reproductive-and-Respiratory-Syndrome (PRRS) was very serious. The financial loss incurred from FMD alone which occurred on Nov. 2011 in Korea was estimated at 3 billion won, 23 % of annual livestock industry production. The livestock industry in Korea has greater risk of disease infection because of high density production, etc. Investigating the spread of livestock diseases should consider both direct and indirect contact as well as other various factors including airborne. Airborne infection of livestock disease was first hypothesised in the early 1900s, however, field experimental studies are still limited. Furthermore, no protocol is available in detecting airborne viruses in the field. In this study, effective virus samplers were investigated by comparative analysis of the type of samplers used detect to airborne virus. Laboratory experiments were conducted to compare virus samplers such as Bio-sampler, Dust-sampler, Compact-Cascade-Impactor (CCI) and Microflow in detecting PRRSV. Samples were analyzed by Reverse-Transcription PCR to assess the efficiency of the instrument in detecting the airborne virus. First, samples were classified into five levels according to light intensity of gel images and then the classified results were normalized. In every case, Bio-sampler and Dust-sampler were comparable with each other and have shown to be more effective than CCI and Microflow samplers.

• 한국지리정보학회지
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• 제18권3호
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• pp.89-101
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• 2015

본 연구는 질병역학의 관점에서 2014년 발생한 HPAI(H5N8)의 시간적 분포와 공간적 분포 그리고 시 공간을 동시에 고려한 분포를 지리정보시스템과 연계하여 분석함으로써 2014년 발생한 HPAI의 전파 및 확산 특징을 알아보고자 한다. 분석 결과 2014년 HPAI는 시간적으로는 모두 3 번의 파동을 형성하였으며, 공간적으로는 경기도 충청북도 충청남도가 인접하는 지역, 전라북도의 곰소만 일대, 전라남도의 영암과 나주 등 영산강과 인접한 지역에서 높은 밀도를 보였다. 시 공간적으로도 공간 밀도가 높은 충청북도 음성지역, 전라북도 부안 고창지역, 나주지역에서 군집이 형성되었다. 다만, 충청북도 음성 진천, 충청남도 천안, 경기도 안성 이천 지역과 전라남도 영암 지역에서는 공간적인 밀도는 높음에도 불구하고 시간적인 범위가 넓음으로써 시 공간 군집이 형성되지 못하였다. 이는 이들 지역의 방역에 문제가 있음을 의미한다. 반면에 곰소만과 인접하고 있는 전라북도 부안 고창 장수 지역은 시 공간 군집이 형성됨으로써, 상대적으로 효과적인 방역이 수행되었다고 볼 수 있다.

• 한국임상수의학회지
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• 제36권1호
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• pp.7-14
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• 2019

In South Korea, six large outbreaks of highly pathogenic avian influenza (HPAI) have occurred since the first confirmation in 2003 from chickens. For the past 15 years, HPAI outbreaks have become an annual phenomenon throughout the country and has extended to wider regions, across rural and urban environments. An understanding of the spatial epidemiology of HPAI occurrence is essential in assessing and managing the risk of the infection; however, local spatial variations of relationship between HPAI incidences in Korea and related risk factors have rarely been derived. This study examined whether spatial heterogeneity exists in this relationship, using a geographically weighted Poisson regression (GWPR) model. The outcome variable was the number of HPAI-positive farms at 252 Si-Gun-Gu (administrative boundaries in Korea) level notified to government authority during the period from January 2014 to April 2016. This response variable was regressed to a set of sociodemographic and topographic predictors, including the number of wild birds infected with HPAI virus, the number of wintering birds and their species migrated into Korea, the movement frequency of vehicles carrying animals, the volume of manure treated per day, the number of livestock farms, and mean elevation. Both global and local modeling techniques were employed to fit the model. From 2014 to 2016, a total of 403 HPAI-positive farms were reported with high incidence especially in western coastal regions, ranging from 0 to 74. The results of this study show that local model (adjusted R-square = 0.801, AIC = 954.5) has great advantages over corresponding global model (adjusted R-square = 0.408, AIC = 2323.1) in terms of model fitting and performance. The relationship between HPAI incidence in Korea and seven predictors under consideration were significantly spatially non-stationary, contrary to assumptions in the global model. The comparison between global Poisson and GWPR results indicated that a place-specific spatial analysis not only fit the data better, but also provided insights into understanding the non-stationarity of the associations between the HPAI and associated determinants. We demonstrated that an empirically derived GWPR model has the potential to serve as a useful tool for assessing spatially varying characteristics of HPAI incidences for a given local area and predicting the risk area of HPAI occurrence. Considering the prominent burden of HPAI this study provides more insights into spatial targeting of enhanced surveillance and control strategies in high-risk regions against HPAI outbreaks.

• Journal of Veterinary Science
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• 제25권2호
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• pp.20.1-20.15
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• 2024

Background: Avian influenza (AI) is a contagious disease that causes illness and death in poultry and humans. High pathogenicity AI (HPAI) H5N6 outbreaks commonly occur in Quang Ninh province bordering China. In June 2021, the first HPAI H5N8 outbreak occurred at a Quang Ninh chicken farm. Objectives: This study examined the risk factors associated with HPAI H5N6 and H5N8 outbreaks in Quang Ninh. Methods: A retrospective case-control study was conducted in Quang Ninh from Nov 2021 to Jan 2022. The cases were households with susceptible poultry with two or more clinical signs and tested positive by real-time reverse transcription polymerase chain reaction. The controls were households in the same village as the cases but did not show clinical symptoms of the disease. Logistic regression models were constructed to assess the risk factors associated with HPAI outbreaks at the household level. Results: There were 38 cases with H5N6 clade 2.3.4.4h viruses (n = 35) and H5N8 clade 2.3.4.4b viruses (n = 3). Compared to the 112 controls, raising poultry in uncovered or partially covered ponds (odds ratio [OR], 7.52; 95% confidence interval [CI], 1.44-39.27), poultry traders visiting the farm (OR, 8.66; 95% CI, 2.7-27.69), farms with 50-2,000 birds (OR, 3.00; 95% CI, 1.06-8-51), and farms with ≥ 2,000 birds (OR, 11.35; 95% CI, 3.07-41.94) were significantly associated with HPAI outbreaks. Conclusions: Combining biosecurity measures, such as restricting visitor entry and vaccination in farms with more than 50 birds, can enhance the control and prevention of HPAI in Quang Ninh and its spread across borders.

• 농촌계획
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• 제18권4호
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• pp.79-89
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• 2012

농가를 방문하는 가금관련업체의 관계자 및 차량은 HPAI 질병 확산의 매개체가 된다. 농가들의 가금관련업체 이용 정보를 이용하면 농가간의 연결을 확인할 수 있고 HPAI 확산 가중 네트워크를 구성할 수 있다. 네트워크 분석중 중심성 측정은 질병에 취약하거나 타 농가에 영향력이 큰 역할을 하는 농가를 분석하는 방법으로 HPAI 초기 확산을 통제하는 방법으로 이용된다. 단, HPAI 바이러스는 네트워크의 연결선 가중치에 따라서 확산 경로가 달라질 수 있다. 기존의 분석 방법은 확산 경로에 있어 대치되는 연결선의 강도와 연결선의 수 중 하나만을 고려하기 때문에 질병 확산을 정확히 모의하는데 한계가 있다. 그래서 본 연구에서는 2008년 발병한 한국 김제 지역의 39개 농가를 대상으로 가금관련업체 이용자료를 적용한 HPAI 확산 네트워크에 연결선의 가중치에 지수를 적용하는 방법으로 기존의 방법과 결과를 비교했다. 이 자료는 가금 산업 네트워크의 한국 지역 농가 적용성을 평가 할 수 있을뿐만 아니라 추후 잠재적인 질병 발병 차단을 위한 정보 제공에 중요한 역할을 할 것이다.

• Animal Bioscience
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• 제36권6호
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• pp.851-860
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• 2023

Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.