Ha-Na Youn

Intranasal administration of live Lactobacillus species facilitates protection against influenza virus infection in mice.

Influenza virus infections continue to be a significant public health problem. For improved therapies and preventive measures against influenza, there has been an increased tendency in modern medicine involving the use of probiotics. In this study, we compared the protective efficacy of various live and dead Lactobacillus species against challenge with influenza virus in mice according to the administration route and dose. In addition, to understand the underlying mechanism behind this clinical protective effect, we performed immunologic assays including examination of IgA levels and cytokine profiles in the lung. The survival rate of mice receiving intranasal administration of Lactobacillus was higher than after oral administration, and administration of live bacteria was more protective than of dead bacteria. The lung levels of interleukin (IL)-12 and IgA were significantly increased (P<0.05). Conversely, the levels of the pro-inflammatory cytokines tumor necrosis factor-alpha and IL-6 were decreased. Interestingly, there were huge differences in protective effects of various Lactobacillus strains on influenza virus infection. Therefore, for clinical applications, selection of effective strains could be critical and individually optimized application regimens of the selected strains are required.

An emerging recombinant cluster of nephropathogenic strains of avian infectious bronchitis virus in Korea.

Abstract The infectious bronchitis virus (IBV) is continuously evolving through point mutation and recombination of their genome, subsequently the emergence of IBV variants complicates disease control. The objective of this study was to investigate genetic characterization of new IBV variants isolated from commercial chicken flocks in Korea collected between 2005 and 2010. Phylogenetic analysis revealed that all new IBV isolates belonged to Korean group II (K-II), which included the nephropathogenic IBV strains. However, the isolates formed a new gene cluster that was distinguished from the two distinct K-II subgroups (KM91-like and QX-like). Recombination events were identified in the S1 gene, with their putative parental strains being the KM91-like or QX-like subgroup. In addition, two crossover sites were observed in the S1 gene of IBV isolates. These results suggest that natural genetic recombination between heterologous strains classified into different genetic groups has occurred and may have caused the emergence of new IBV strains. This finding provides important information on IBV evolution and is essential for the effective control of IB in Korea.

Continuing evolution and interspecies transmission of influenza viruses in live bird markets in Korea.

Abstract Live bird markets (LBMs) provide an ideal environment for the evolution and interspecies transfer of avian influenza viruses (AIVs). In this study, we analyzed AIVs present in LBMs in Korea during the winter seasons of 2006–08. Sixty-five AIVs that belong to four hemagglutination (HA) subtypes of AIV (H3, H4, H6, and H9) were isolated from 644 pooled tissue or swab samples collected in LBMs. Most H9 subtypes of AIVs were isolated from Galliformes (chickens, silky fowls, pheasants, and guinea fowls), and other subtypes were isolated from Anseriformes (Pekin ducks and mallards). In addition, we obtained a single H3N2 virus from nasal swabs of dogs sold in LBMs, and the virus was genetically identical to the canine influenza virus (CIV) isolated from pet dogs in Korea. Phylogenetic analysis suggests that the Korean H9N2 viruses prevalent in chickens have provided their gene segments to AIVs circulating in ducks. These gene transfers facilitated reassortment events among AIVs and likely generated the ancestors of CIV in Korea. An animal challenge study using chickens, quail, mice, and dogs had shown that the H4 and H6 subtypes could replicate in mice and that some H4 and H6 viruses could replicate in chickens without preadaptation. In addition, two H3 subtype viruses (H3N2 and H3N8) induced interstitial pneumonia that accompanied clinical signs and seroconversion in dogs. Our findings indicate that the newly evolved AIVs have been continuously generated by reassortment in ducks, and these reassortments could result in expanding the host range of AIVs.

An inactivated oil-emulsion fowl Adenovirus serotype 4 vaccine provides broad cross-protection against various serotypes of fowl Adenovirus.

The number of clinical cases of inclusion body hepatitis (IBH) and hydropericardium-hepatitis syndrome (HHS) has been increasing, resulting in considerable economic losses in many countries. Currently, only fowl Adenovirus (FAdV) serotype 4 (FAdV-4) has been reported as the causative agent of HHS, whereas IBH can be caused by all 12 serotypes of FAdV. For protection against HHS, various live and killed FAdV serotype 4 vaccines have been developed. However, there is a concern whether these vaccines composed of FAdV-4 alone could provide protection against IBH, which is caused by other serotypes of virulent FAdVs. To date, there have been no reports evaluating the protective efficacy of the FAdV-4 vaccine against other serotypes of FAdV. Thus, we investigated the cross-protection efficacy of an inactivated oil-emulsion FAdV-4 vaccine against various serotypes of FAdV field isolates. Our study demonstrated that the inactivated oil-emulsion FAdV-4 vaccine could provide broad cross-protection against various serotypes of FAdV in not only vaccinated birds, but also the progenies of vaccinated breeder. Therefore, we conclude that the inactivated oil-emulsion FAdV-4 vaccine could be effective in preventing the spread of various other serotypes of FAdV as well as FAdV-4 infection in the poultry industry.

Identification and virulence characterization of fowl adenoviruses in Korea.

SUMMARY. Since 2007, 55 adenovirus strains have been isolated from commercial chicken flocks in Korea and have been identified and the pathogenicity of these isolates was confirmed in specific-pathogen-free chickens of different age. Based on sequencing analysis of the hexon gene, 55 FAdV isolates were genetically related to the IBH-2A strain of FAdV3 (4 isolates, 99.2% to 100%), the KR5 strain of FAdV4 (22 isolates, 97.9% to 99.2%), the 764 strain of FAdV9 (11 isolates, 99.1% to 99.3%), and the 1047 strain of FAdV11 (18 isolates, >99%). Experimental infections with four serotypes of FAdV resulted in high mortality of 18-day-old chicken embryos and 1-day-old chicks with marked liver necrosis similar to those observed in the natural outbreaks. Notably, specific hydropericardium was observed in chicks challenged with the K531 strain (serotype 4). However, 3-wk-old chickens challenged with FAdVs, regardless of serotype, did not show any clinical signs or mortality except histologic lesions of focal hepatocytic necrosis with mild lymphocytic infiltration. The results indicate that four FAdV serotypes (3, 4, 9, and 11) are the dominant serotypes of FAdVs in the Korea and are pathogenic enough to cause clinical disease in young chicks. The present investigation provides important information on the epidemiology and pathogenesis of FAdVs and highlights the importance of control strategies against FAdV infection in Korea.

Use of bacteriophage for biological control of Salmonella Enteritidis infection in chicken

Bacteriophage ΦCJ07 with broad host ranges for Salmonella strains isolated from sewage effluent were used to reduce Salmonella Enteritidis (SE) infection in chickens. One-day-old chicks challenged with 5×10(7) colony-forming units/bird of SE were cohabitated with contact chicks and treated with three concentrations (10(5), 10(7) and 10(9) plaque forming units (PFU)/g) of bacteriophage prepared as a feed additive for 21days after challenge. Salmonella in the intestine was quantified and environmental contamination level was examined at 1, 2 and 3weeks after challenge. All treatments reduced intestinal SE colonization in challenged and contact chickens and reduced the environmental contamination level, but the reductions produced by 10(7) and 10(9)PFU/g of bacteriophage were significant (P<0.05) as compared with untreated controls. In addition, seven out of 10 (70%) contact chickens treated with 10(9)PFU/g of bacteriophage had no detectable intestinal Salmonella at 3weeks after treatment, suggesting that bacteriophage therapy significantly prevented the horizontal transmission of SE. These results provide important insights into preventive and control strategies against SE infection in poultry and indicate that the use of bacteriophage could reduce the incidence of Salmonella food poisoning.

Sublingual administration of Lactobacillus rhamnosus affects respiratory immune responses and facilitates protection against influenza virus infection in mice.

The extensive morbidity and mortality caused by influenza A viruses worldwide prompts the need for a deeper understanding of the host immune response and novel therapeutic and/or prophylactic interventions. In this study, we assessed the sublingual route as an effective means of delivering probiotics against influenza virus in mice. In addition, IgA levels, NK cell activity, T cell activation, and cytokine profiles in the lungs were examined to understand the mechanism underlying this protective effect. Sublingual administration of Lactobacillus rhamnosus provided enhanced protection against influenza virus infection by enhancing mucosal secretory IgA production, and T and NK cell activity. Moreover, interleukin (IL)-12 levels in the lungs increased significantly. Conversely, IL-6 and tumor necrosis factor alpha levels in the lungs decreased significantly. On the basis of these promising findings, we propose that the sublingual mucosal route is an attractive alternative to mucosal routes for administering probiotics against influenza virus.

Prevalence and characterization of Salmonella species in entire steps of a single integrated broiler supply chain in Korea

The purpose of this study was to investigate the distribution of Salmonella species in an integrated broiler supply chain in Korea. A total of 1,214 samples from various steps of an integrated broiler production company including broiler breeder farms, broiler farms, broiler trucks, slaughterhouse, and retail chicken meats were collected and investigated. Salmonella was detected in 195 of the samples. The highest prevalence of Salmonella was observed in broiler transporting trucks (71.43%), followed by the slaughterhouse (63.89%) and broiler farms (16.05%). Salmonella Hadar was the most frequently isolated serotype (83.08%). All Salmonella Hadar isolates investigated in this study with pulsed-field gel electrophoresis showed the same XbaI pulsed-field gel electrophoresis pulsotype.

Evidence of intercontinental transfer of North American lineage avian influenza virus into Korea.

Avian influenza viruses (AIV) can be genetically distinguished by geographical origin. The present study found evidence of intercontinental transfer of North American lineage AIV into Asia via migratory bird populations. The North American lineage genes were detected in live animal markets during avian influenza surveillance, seemed to have reassorted with Eurasian AIV in wild bird habitats, and had transmitted to live animal markets. Enhanced AIV surveillance is required to understand the influence of newly transferred North American lineage AIV genes on AIV evolution in Asia and to investigate AIV ecology in various transcontinental migrant species.

Cross-Protective Immune Responses Elicited by a Korean Variant of Infectious Bronchitis Virus

SUMMARY Infectious bronchitis virus (IBV) infections cause great economic losses to the poultry industry worldwide. IBVs continuously evolve by developing mutations in antigenic sites; therefore, an IBV vaccine that provides broad cross-protection can be a highly relevant and practical method in IBV control strategies. Although some IBV vaccine strains are known to provide protection against multiple IBV serotypes, in general commercially available IBV vaccine strains provide protection against antigenically related viruses but not distinct heterologous viruses. In the present study we characterized the Korean variant IBV K40/09 strain with regard to its immunogenicity and protective efficacy against seven currently circulating IBV serotypes. Three-week-old specific-pathogen-free chickens were intraocularly immunized with the IBV K40/09 strain at 103.5 50% egg infective dose (EID50). Three weeks after immunization all the birds were challenged with seven different strains at 104.5 EID50. Chickens immunized with the IBV K40/09 strain showed significantly high levels of protection against all challenge viruses at the trachea and kidney levels. Our results suggest that IBV K40/09 could be useful to ensure IBV vaccine effectiveness owing to its cross-protective ability. Therefore, the IBV K40/09 strain merits consideration as a vaccine candidate to prevent infection as well as the spread of new IBV strains and many IBV variants that have been reported worldwide. RESUMEN Nota de Investigación—Respuesta inmune protectora cruzada provocada por una variante coreana del virus de la bronquitis infecciosa. Las infecciones por el virus de bronquitis infecciosa (IBV) causan grandes pérdidas económicas a la industria avícola en todo el mundo. Los virus de la bronquitis infecciosa evolucionan continuamente mediante el desarrollo de mutaciones en sitios antigénicos, por lo tanto, una vacuna contra el virus de bronquitis infecciosa que proporciona una amplia protección cruzada puede ser un método altamente relevante y práctico dentro de las estrategias de control contra este virus. Aunque algunas cepas vacunales del virus de bronquitis proporcionan protección contra múltiples serotipos del virus, en general, las cepas vacunales disponibles comercialmente proporcionan protección contra virus antigénicamente relacionados pero no contra virus heterólogos distintos. En el presente estudio se caracterizó la cepa variante coreana IBV K40/09 con relación a su inmunogenicidad y eficacia protectora contra siete serotipos que circulan actualmente. Aves libres de patógenos específicos de tres semanas de edad, fueron inmunizadas por vía intraocular con la cepa IBV K40/09 con 103.5 dosis infectantes en embrión de pollo 50% (EID50). Tres semanas después de la vacunación, todas las aves fueron desafiadas con siete cepas diferentes, con 104.5 EID50. Los pollos inmunizados con la cepa K40/09 mostraron niveles significativamente altos de protección contra todos los virus de desafío en la tráquea y en riñón. Nuestros resultados sugieren que la cepa IBV K40/09 podría ser útil y eficaz como vacuna contra el virus de bronquitis infecciosa debido a su capacidad de protección cruzada. Por lo tanto, la cepa K40/09 merece consideración como un candidato a vacuna para prevenir la infección y la propagación de nuevas cepas del virus de la bronquitis infecciosa y variantes que han sido reportadas en todo el mundo.