• Korean Journal of Veterinary Service
• /
• v.40 no.1
• /
• pp.15-20
• /
• 2017

A two-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was designed for the rapid visual detection of the M gene of all subtypes of avian influenza virus (AIV) and the H5 gene of the H5 subtype of highly pathogenic AIV (HPAIV). The reaction carried out in two tubes in a single step at $58^{\circ}C$ for 40 min, and the assay results could be visually detected by using hydroxynaphthol blue dye. Using M or H5 gene-specific primers, the assay successfully detected all subtypes or H5 subtypes of AIVs, including the Korean representative H5N1 and H5N8 HPAIVs. The detection limit of the assay was approximately $10^{2.0}$ $EID_{50}/reaction$ for the M and H5 genes of H5N1 HPAIV, respectively, and was more sensitive than that of previously reported RT-LAMP and comparable to that of real-time RT-PCR. These results suggest that the present RT-LAMP assay, with its high specificity, sensitivity, and simplicity, will be a useful diagnostic tool for surveillance of currently circulating H5 HPAIVs and other subtypes of AIV in bird population, even in under-equipped laboratories.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.1
• /
• pp.151-159
• /
• 2016

Nanoparticles have wide-scale applications in various areas, including medicine, chemistry, electronics, and energy generation. Several physical, biological, and chemical methods have been used for synthesis of silver nanoparticles. Green synthesis of silver nanoparticles using plants provide advantages over other methods as it is easy, efficient, and eco-friendly. Nanoparticles have been extensively studied as potential antimicrobials to target pathogenic and multidrug-resistant microorganisms. Their applications recently extended to development of antivirals to inhibit viral infections. In this study, we synthesized silver nanoparticles using Cinnamomum cassia (Cinnamon) and evaluated their activity against highly pathogenic avian influenza virus subtype H7N3. The synthesized nanoparticles were characterized using UVVis absorption spectroscopy, scanning electron microscopy, and Fourier transform infrared spectroscopy. Cinnamon bark extract and its nanoparticles were tested against H7N3 influenza A virus in Vero cells and the viability of cells was determined by tetrazolium dye (MTT) assay. The silver nanoparticles derived from Cinnamon extract enhanced the antiviral activity and were found to be effective in both treatments, when incubated with the virus prior to infection and introduced to cells after infection. In order to establish the safety profile, Cinnamon and its corresponding nanoparticles were tested for their cytotoxic effects in Vero cells. The tested concentrations of extract and nanoparticles (up to 500 μg/ml) were found non-toxic to Vero cells. The biosynthesized nanoparticles may, hence, be a promising approach to provide treatment against influenza virus infections.

• Korean Journal of Microbiology
• /
• v.54 no.3
• /
• pp.254-265
• /
• 2018

Avian influenza recently damaged the poultry industry, which suffered a huge economic loss reaching billions of U.S. dollars in South Korea. Transmission routes of the pathogens would help plan to control and limit the spread of the devastating biological tragedy. Phylogenetic analyses of pathogen's DNA sequences could sketch transmission trees relating hosts with directed edges. The last decade has seen the methodological development of inferring transmission trees using epidemiological as well as genetic data. Here, I reanalyzed the DNA sequence data that had originated in the highly pathogenic avian influenza H5N8 outbreak of South Korea in 2014. The H5N8 viruses spread geographically contiguously from the origin of the outbreak, Jeonbuk. The Jeonbuk origin viruses were known to spread to four provinces neighboring Jeonbuk. I estimated the transmission tree of the host domestic and migratory wild birds after combining multiple runs of Markov chain Monte Carlo using a Bayesian method for inferring transmission trees. The estimated transmission tree, albeit with a rather large uncertainty in the directed edges, showed that the viruses spread from Jeonbuk through Chungnam to Gyeonggi. Domestic birds of breeder or broiler ducks were estimated to appear to be at the terminal nodes of the transmission tree. This observation confirmed that migratory wild birds played an important role as one of the main infection mediators in the avian influenza H5N8 outbreak of South Korea in 2014.

• Journal of Veterinary Science
• /
• v.24 no.5
• /
• pp.73.1-73.16
• /
• 2023

Background: Highly pathogenic avian influenza virus (HPAIV) is considered a global threat to both human health and the poultry industry. MicroRNAs (miRNA) can modulate the immune system by affecting gene expression patterns in HPAIV-infected chickens. Objectives: To gain further insights into the role of miRNAs in immune responses against H5N1 infection, as well as the development of strategies for breeding disease-resistant chickens, we characterized miRNA expression patterns in tracheal tissues from H5N1-infected Ri chickens. Methods: miRNAs expression was analyzed from two H5N1-infected Ri chicken lines using small RNA sequencing. The target genes of differentially expressed (DE) miRNAs were predicted using miRDB. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were then conducted. Furthermore, using quantitative real-time polymerase chain reaction, we validated the expression levels of DE miRNAs (miR-22-3p, miR-146b-3p, miR27b-3p, miR-128-3p, miR-2188-5p, miR-451, miR-205a, miR-203a, miR-21-3p, and miR-200a3p) from all comparisons and their immune-related target genes. Results: A total of 53 miRNAs were significantly expressed in the infection samples of the resistant compared to the susceptible line. Network analyses between the DE miRNAs and target genes revealed that DE miRNAs may regulate the expression of target genes involved in the transforming growth factor-beta, mitogen-activated protein kinase, and Toll-like receptor signaling pathways, all of which are related to influenza A virus progression. Conclusions: Collectively, our results provided novel insights into the miRNA expression patterns of tracheal tissues from H5N1-infected Ri chickens. More importantly, our findings offer insights into the relationship between miRNA and immune-related target genes and the role of miRNA in HPAIV infections in chickens.

• Korean Journal of Veterinary Research
• /
• v.47 no.4
• /
• pp.399-407
• /
• 2007

Cause of high lethality and dissemination to human being, new development of rapid method for the detection of highly pathogenic Avian Influenza Virus (AIV) is still necessary. For the detection of AIV subtype H5N1, typical pathogenic AIV, new method to confirm sub-typing of this virus is also needed. For the purpose of ultra-rapid detection and sub-typing of hemagglutinin and neuraminidase of AIV, this study was planned. As the results we could demonstrate an ultra-rapid multiplex real-time PCR (URMRT PCR) for the detection of AIV In this study, the URMRT PCR were optimized with synthesized AIV H5- and AIV Nl-specific DNA templates and GenSpector TMC, which is a semiconductor process technology based real-time PCR system with high frequencies of temperature monitoring. Under eight minutes, the amplifications of two AIV subtype-specific PCR products were successfully and independently detected by 30 cycled ultra-rapid PCR, including melting point analysis, from $1{\times}10^3$ copies of mixed template DNA. The URMRT PCR for the detection of AIV H5N 1 developed in this study could be expected to apply not only detections of different AIVs, but also various pathogens. It was also discussed that this kind of the fastest PCR based detection method could be improved by advance of related technology in near future.

• The Journal of Bigdata
• /
• v.5 no.1
• /
• pp.1-9
• /
• 2020

The study was conducted with funding from the government (Ministry of Agriculture, Food and Rural Affairs) in 2018 with support from the Agricultural, Food, and Rural Affairs Agency, 318069-03-HD040, and in based on artificial intelligence-based HPAI spread analysis and patterning. The model that is actively used in time series and text mining recently is LSTM (Long Short-Term Memory Models) model utilizing deep learning model structure. The LSTM model is a model that emerged to resolve the Long-Term Dependency Problem that occurs during the Backpropagation Through Time (BPTT) process of RNN. LSTM models have resolved the problem of forecasting very well using variable sequence data, and are still widely used.In this paper study, we used the data of the Call Detailed Record (CDR) provided by KT to identify the migration path of people who are expected to be closely related to the virus. Introduce the results of predicting the path of movement by learning the LSTM model using the path of the person concerned. The results of this study could be used to predict the route of HPAI propagation and to select routes or areas to focus on quarantine and to reduce HPAI spread.

• Journal of Veterinary Science
• /
• v.23 no.2
• /
• pp.24.1-24.10
• /
• 2022

Background: Small interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia. Objectives: The main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus. Methods: The effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells. Results: The siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene. Conclusions: These findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.

• Korean Journal of Poultry Science
• /
• v.46 no.4
• /
• pp.313-325
• /
• 2019

In most cases of HPAI (highly pathogenic avian influenza) outbreaks, stamping-out operations are initiated by officially designating the affected premise, which is subsequently followed by depopulation of infected flocks. The primary objective of this study was to develop an evaluation method that correlates the level of biosecurity and the risk of having an HPAI outbreak in domestic poultry farms. A total of eight farms were selected nationwide, including layer farms, broiler farms, and an animal welfare type farm. The biosecurity level of the chosen poultry farms was assessed based on a total scoring index of 183 divided into three categories, general management (51), quarantine management (106), and sanitation management (26). Conclusively, the five layer farms (JS, GE, CS, HS, OE), scored higher overall scores compared to the animal welfare farm (CH) and broiler farms (JG, LB). In terms of scoring, which adds up to a total of 183 points, most layer farms scored between 130 and 157, while the two broiler farms and the welfare farm scored 45, 75 and 70, respectively. Next, an independent HPAI risk assessment of the farms was carried out. Regarding the correlation between biosecurity levels and HPAI risks, in the farms that presented a higher overall score in terms of biosecurity and outweighed the risks of HPAI, they tended to earn more points in the quarantine management category. The results of this study suggest that a viable system for evaluating biosecurity levels can establish strong correlations with the risk of having HPAI.

• Korean Journal of Veterinary Research
• /
• v.59 no.2
• /
• pp.97-100
• /
• 2019

It is important to select the correct disinfectants and to use them appropriately in order to prevent the initial spread of highly infectious livestock disease, such as foot-and-mouth disease or highly pathogenic avian influenza. This study describes a smartphone application developed to enable livestock workers to observe information related to disinfectants for the prevention of livestock disease in the domestic market, regardless of time and location, through a Linux-based Android mobile platform. This application (Konkuk-Disinfectant Information Database) provides information on disinfectant names, components, license and use; it was designed to enable the user to share disinfectant information through social media.

• Journal of Food Hygiene and Safety
• /
• v.27 no.1
• /
• pp.18-23
• /
• 2012

Highly pathogenic avian influenza virus (HPAIV) is already panzootic in poultry and caused a considerable economic loss in poultry industry. In addition, HPAIV continues to cross species barriers to infect humans and other mammals, often with fatal outcomes. In this study, the virucidal efficacy of Citra-$Kill^{(R)}$ composed to quaternary ammonium chloride and citric acid was investigated against avian influenza H9N2 virus (AIV). A virucidal efficacy was determined with the viability of AIV contacted with the disinfectant in the allantoic membrane of chicken embryos. Citra-$Kill^{(R)}$ and AIV was reacted on the distilled water (DW), hard water (HW) or organic matter suspension (OM) condition. On DW condition, AIV was inactivated with 2,000 fold dilutions of Citra-$Kill^{(R)}$. When the antiviral effect on HW condition was evaluated, the antiviral activity of the disinfectant showed on 1,500 fold dilutions against AIV. With the investigation of the antiviral effect of the disinfectant on OM condition, AIV was inactivated on 500 fold dilutions of Citra-$Kill^{(R)}$. As Citra-$Kill^{(R)}$ possesses virucidal efficacy against AIV, the disinfectant solution can be used to limit the spread of animal viral diseases.