• Journal of Veterinary Science
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• 제23권2호
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• pp.24.1-24.10
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• 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.

• 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.

• BMB Reports
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• 제40권6호
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• pp.1002-1008
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• 2007

PreS domain of Hepatitis B virus (HBV) surface antigen is a good candidate for an effective vaccine as it activates both B and T cells besides binding to hepatocytes. This report deals with overexpression and purification of adr subtype of surface antigen that is more prevalent in Pakistan. PreS region, comprising 119 aa preS1 region plus a 55 aa preS2 region plus 11 aa from the N-terminal S region, was inserted in pET21a+ vector, cloned in E. coli $DH5\alpha$ cells and expressed in E. coli BL21 codon+ cells. The conditions for over expression were optimized using different concentrations of IPTG (0.01-5 mM), and incubating the cells at different temperatures (23-$41^{\circ}C$) for different durations (0-6 h). The cells were grown under the given optimized conditions (0.5 mM IPTG concentration at $37^{\circ}C$ for 4 h), lysed by sonication and the protein was purified by ion exchange chromatography. On the average, 24.5 mg of recombinant protein was purified per liter of culture. The purified protein was later lyophilized and stored at $-80^{\circ}C$.

• 대한수의학회지
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• 제44권3호
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• pp.449-454
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• 2004

Ninety pigs under the age of 120-day-old requested at the diagnostic laboratory of animal diseases in Cheju National University were evaluated for the prevalence of tissue antigen and serum antibody to swine influenza virus (SIV). For histopathologic examination there was sampled at the consolidated area in cranioventral or dorsocaudal lobes of lungs. Lung tissues from all pigs were tested for the antigen of SIV type A by immunohistochemistry (IHC). Sera from 56 pigs were used for the antibody detection to SIV type A (subtype H1N1 and H3N2) by haemagglutinin inhibition test. Pneumonic lesions were observed in 72 cases (80%) of 90 pigs. Broncho-interstitial or interstitial pneumonia were more prevalent than suppurative or fibrinous bronchopneumonia. According to HI test, 46.4% of the tested sera showed seropositive. Positive sera were consisted with 5.3% for SIV H1N1, 28.6% for SIV H3N2, and 12.5% for both subtype to be tested, respectively. SIV antigens were detected in 51 cases(56.6%) of 90 pigs. Most SIV antigens were presented in the epithelium of the bronchi and bronchiole. Necrotizing bronchitis or bronchiolitis were observed in 28(31.1%) cases of all inspected pigs. These results suggested that SIV might be an important role to induce swine pneumonia in Jeju. Also IHC was very useful for the detection of SIV in the lung.

• 대한간호학회지
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• 제41권3호
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• pp.403-410
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• 2011

Purpose: This study was to identify the current status of vaccination against the pandemic (H1N1) 2009 virus among university students from the fields of nursing and allied health from a local community and verify factors influencing vaccination. Methods: The study included 227 students in the fields of nursing and allied health from a provincial university. Data were obtained from these participants between May 31 and June 11, 2010 by using self-report questionnaires. Results: The rate of vaccination against the pandemic (H1N1) 2009 virus for these participants was 14.5%. No difference was observed in this regard between majors and school year. Factors that influence vaccination against this virus included previous vaccination against seasonal influenza and participants’ attitudes toward general vaccination. Conclusion: The results suggest that for effective pandemic (H1N1) 2009 vaccination of university students from the fields of nursing and allied health, students who have not been vaccinated should be intensively managed. Developing a vaccination program that encourages a positive attitude toward vaccination is recommended.

• 한국가금학회지
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• 제40권3호
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• pp.243-252
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• 2013

Among the 16 hemagglutinin (HA) subtypes of avian influenza virus (AIV), only the H5 and H7 subtypes have caused highly pathogenic avian influenza (HPAI) in poultry. However, most H5 or H7 subtype viruses are categorized as low pathogenic avian influenza (LPAI). Some AIVs, including the H5 and H7 HPAI viruses, have shown the ability to infect humans directly. In this study, we describe the biological and molecular characterization of an H5N3 AIV (SBD/KR/KNU SYG06/06) isolated from spot-billed duck (Anas poecilorhyncha) in Korea. A phylogenetic analysis of the eight viral genes showed that the SBD/KR/KNU SYG06/06 isolate belongs to the Eurasian lineage and that the SBD/KR/KNU SYG06/06 isolate was clearly different from HPAI H5N1 strains, including human isolates and the Italian HPAI H5N2 strains. Additionally, no relationship was found between SBD/KR/KNU SYG06/06 and the Korean HPAI H5N1 isolates. The SBD/KR/ KNU SYG06/06 isolate had avian specific receptor binding site residues in the HA protein and the four C-terminal amino acids in the NS1 protein. The HA protein of the SBD/KR/KNU SYG06/06 isolate exhibited the typical LPAI motif at the cleavage site and this virus produced no cytopathic effects in MDCK cells without trypsin. Given these results, we suggest that the H5N3 AIV isolated from the spot-billed duck should be considered an LPAI virus and should have no pathogenic effect in humans.

• 생명과학회지
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• 제17권3호통권83호
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• pp.340-344
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• 2007

조류인플루엔자바이러스는 사람 및 동물에 상당한 위협을 가하고 있다. 이 연구는 알칼리성 소독용액이 H5N1, H3N2, H6N1, H9N2형 조류인플루엔자바이러스를 불활성화시킬 수 있는지를 조사했다. 알칼리성 소독용액을 생리식염수에 100배 희석한 경우 조류인플루엔자가 MDCK 세포에서 증식하는 것을 완전히 억압하였다. 형광항체법을 이용한 실험에서도 알칼리성소독액을 처리한 H9N2 조류인플루엔자바이러스는 MDCK세포에서 증식이 억제되었고, 알칼리성 소독액을 처리하지 않은 H9N2조류인플루엔자바이러스는 증식이 억제되지 않았다. 이 결과는 알칼리성 소독액은 고병원성 H5N1을 포함을 조류인플루엔자바이러스를 방역하는데 도움이 될 수 있음을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제18권6호
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• pp.1164-1169
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• 2008

We developed multiplex RT-PCR assays that can detect and identify 12 hemagglutinin (H1-H12) and 9 neuraminidase (N1-N9) subtypes that are commonly isolated from avian, swine, and human influenza A viruses. RT-PCR products with unique sizes characteristic of each subtype were amplified by multiplex RT-PCRs, and sequence analysis of each amplicon was demonstrated to be specific for each subtype with 24 reference viruses. The specificity was demonstrated further with DNA or cDNA templates from 7 viruses, 5 bacteria, and 50 influenza A virus-negative specimens. Furthermore, the assays could detect and subtype up to $10^5$ dilution of each of the reference viruses that had an original infectivity titer of $10^6\;EID_{50}/ml$. Of 188 virus isolates, the multiplex RT-PCR results agreed completely with individual RT-PCR subtyping results and with results obtained from virus isolations. Furthermore, the multiplex RT-PCR methods efficiently detected mixed infections with at least two different subtypes of influenza viruses in one host. Therefore, these methods could facilitate rapid and accurate subtyping of influenza A viruses directly from field specimens.

• Archives of Pharmacal Research
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• 제23권5호
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• pp.518-524
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• 2000

We examined the modulation of protein kinase C (PKC) subtypes during apoptosis induced by ginsenoside Rh2 (G-Rh2) in human neuroblastoma SK-N-Bl(2) and rat glioma C6Bu-1 cells. Apoptosis induced by C-Rh2 in both cell lines was confirmed, as indicated by DNA fragmentation and in situ strand breaks, and characteristic morphological changes. During apoptosis induced by G-Rh2 in SK-N-BE(2) cells, PKC subtypes $\alpha$, $\beta$ and $\gamma$ were progressively increased with prolonged treatment, whereas PKC $\delta$ increased transiently at 3 and 6 h and PKC $\varepsilon$ was gradually down-regulated after 6 h following the treatment. On the other hand, PKC subtype $\beta$ markedly increased at 24 h when maximal apoptosis was achieved. In C6Bu-l cells, no significant changes in PKC subtypes $\alpha$, $\gamma$, $\delta$, $\varepsilon$ and $\beta$ were observed during apoptosis induced by G-Rh2. These results suggest the evidence for a possible role of PKC subtype in apoptosis induced by G-Rh2 in SK-N-BE(2) cells but not in C6Bu-1 cells, and raise the possibility that G-Rh2 may induce apoptosis via different pathways interacting with or without PKC in different cell types.

• 센서학회지
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• 제23권4호
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• pp.278-283
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• 2014

In this study, we propose an immunosensor using zinc oxide nanorods (NRs) inside PDMS channel for detecting the influenza A virus subtype H7N9. ZnO with high isoelectric point (IEP, ~9.5) makes it suitable for immobilizing proteins with low IEP. In this proposed H7N9 immunosensor structure ZnO NRs were grown on the PDMS channel inner surface to immobilize H7N9 capture antibody. A sandwich enzyme-linked immunosorbent assay (ELISA) method with was used 3,3',5,5' tetramethylbenzidine (TMB) for detecting H7N9 influenza virus. The immunosensor was evaluated by amperometry at various H7N9 influenza antigen concentrations (1 pg/ml - 1 ng/ml). The redox peak voltage and current were measured by amperometry with ZnO NWs and without ZnO NWs inside PDMS channel. The measurement results of the H7N9 immunosensor showed that oxidation peak current of TMB at 0.25 V logarithmically increased from 2.3 to 3.8 uA as the H7N9 influenza antigen concentration changed from 1 pg/ml to 1 ng/ml. And then we demonstrated that ZnO NRs inside PDMS channel can improve the sensitivity of immunosensor to compare non-ZnO NRs inside PDMS channel.