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Yesan, South Korea

Cho Y.,Kangwon National University | Park S.,Kangwon National University | Barate A.K.,Kangwon National University | Truong Q.L.,Kangwon National University | And 5 more authors.
Journal of Microbiology and Biotechnology | Year: 2015

Salmonella enterica serovar Enteritidis is the predominant agent causing salmonellosis in chickens and other domestic animals. In an attempt to identify antigenic S. Enteritidis outer membrane proteins (OMPs) that may be useful for subunit vaccine development, we established a proteomic map and database of antigenic S. Enteritidis OMPs. In total, 351 and 301 spots respectively from S. Enteritidis strain 270 and strain 350 were detected by twodimensional gel electrophoresis. Fifty-one antigen-reactive spots were detected by antisera on two-dimensional immunoblots and identified as 12 specific proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. OmpA and DNA starvation/ stationary phase protection protein (Dps) were the most abundant proteins among the identified OMPs, comprising 22 and 12 protein species, respectively. Interestingly, we found that the Dps of S. Enteritidis is also antigenic. OmpW was also verified to have high antigenicity. These results show that OmpA, Dps, and possibly OmpW are antigenic proteins. This study provides new insights into our understanding of the immunogenic characteristics of S. Enteritidis OMPs. © 2015 by The Korean Society for Microbiology and Biotechnology.

Park W.-J.,Konkuk University | Park B.-J.,Konkuk University | Song Y.-J.,Konkuk University | Lee J.-B.,Konkuk University | And 8 more authors.
Japanese Journal of Veterinary Research | Year: 2015

Immunocastration is an alternative method used to replace surgical castration commonly performed in swine farms. In boars, the main effects of immunocastration are reduction of gonadotropin-releasing hormone (GnRH) and the resulting inhibition of testicular function. The aim of this study was to evaluate immunocastration efficacy in pre-pubertal boars vaccinated with a recombinant GnRH protein conjugated with Salmonella Typhimurium flagellin fljB (STF2). A total of 35 boars were assigned to three groups: the untreated group (n = 5), the surgically castrated group (n = 5), and the immunocastrated group (n = 25). Pigs in the immunocastration group were immunized with the GnRH-STF2 vaccine at pre-pubertal ages 4 and 8 weeks. All experimental pigs were kept for 26 weeks before slaughter. AntiGnRH antibody levels of immunocastrated pigs were significantly higher than those of untreated pigs (P < 0.001). In contrast, testosterone levels of immunocastrated pigs were significantly lower than those of untreated pigs (P < 0.001). Statistical significances were not found in the body weights and backfat thicknesses of untreated vs. immunocastrated pigs. Weights of the testes and epididymides of immunocastrated pigs were significantly lower than those of untreated pigs (P < 0.001). Testicular tissues of immunocastrated pigs were severely suppressed compared with those of untreated pigs. In conclusion, immunization with the STF2-GnRH vaccine effectively induced immunocastration in pre-pubertal boars. © 2015, Hokkaido University. All rights reserved.

Lee J.S.,Gyeongsang National University | Jang H.B.,Catholic University of Leuven | Kim K.S.,KBNP Inc. | Kim T.H.,KBNP Inc. | And 5 more authors.
PLoS ONE | Year: 2015

Avian pathogenic Escherichia coli (APEC) is a major pathogen that causes avian colibacillosis and is associated with severe economic losses in the chicken-farming industry. Here, bacteriophage KBNP1315, infecting APEC strain KBP1315, was genomically and functionally characterized. The evolutionary relationships of KBNP1315 were analyzed at the genomic level using gene (protein)-sharing networks, the Markov clustering (MCL) algorithm, and comparative genomics. Our network analysis showed that KBNP1315 was connected to 30 members of the Autographivirinae subfamily, which comprises the SP6-, T7-, P60-, phiKMV-, GAP227-and KP34-related groups. Network decomposition suggested that KBNP1315 belongs to the SP6-like phages, but our comparison of putative encoded proteins revealed that key proteins of KBNP1315, including the tail spike protein and endolysin, had relative low levels of amino acid sequence similarity with other members of the SP6-like phages. Thus KBNP1315 may only be distantly related to the SP6-like phages, and (based on the difference in endolysin) its lysis mechanism may differ from theirs. To characterize the lytic functions of the holin and endolysin proteins from KBNP1315, we expressed these proteins individually or simultaneously in E. coli BL21 (DE3) competent cell. Interestingly, the expressed endolysin was secreted into the periplasm and caused a high degree of host cell lysis that was dose-dependently delayed/blocked by NaN3-mediated inhibition of the SecA pathway. The expressed holin triggered only a moderate inhibition of cell growth, whereas coexpression of holin and endolysin enhanced the lytic effect of endolysin. Together, these results revealed that KBNP1315 appears to use a pin-holin/signal-Arrestrelease (SAR) endolysin pathway to trigger host cell lysis. © 2015 Lee et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Kim Y.-W.,KBNP Inc. | Jeong Y.-J.,KBNP Inc. | Kim A.-Y.,Chonnam National University | Son H.-H.,KBNP Inc. | And 4 more authors.
PLoS ONE | Year: 2014

Five novel Lactobacillus brevis strains were isolated from naturally fermented Aloe vera leaf flesh. Each strain was identified by Random Amplified Polymorphic DNA (RAPD) analysis and 16S rRNA sequence comparison. These strains were highly tolerant to acid, surviving in pH2.5 for up to 4 hours, and resistant to 5% bile salts at 37°C for 18 hours. Due to its tolerance to acid and bile salts, one strain passed through the gastric barrier and colonised the intestine after oral administration. All five strains inhibited the growth of many harmful enteropathogens without restraining most of normal commensals in the gut and hence named POAL (Probiotics Originating from Aloe Leaf) strains. Additionally, each strain exhibited discriminative resistance to a wide range of antibiotics. The L. brevis POAL strains, moreover, expressed high levels of the glutamate decarboxylase (GAD) gene which produces a beneficial neurotransmitter, γ-aminobutyric acid (GABA). These characteristics in all suggest that the novel L. brevis strains should be considered as potential food additives and resources for pharmaceutical research. © 2014 Kim et al.

Sun J.,Kangwon National University | Cho Y.,Kangwon National University | Jang J.-H.,KBNP Inc. | Kang Z.-W.,KBNP Inc. | And 2 more authors.
Korean Journal for Food Science of Animal Resources | Year: 2013

Salmonella Gallinarum (SG) is known as an important pathogen that causes fowl typhoid in chickens. To investigate SG outer-membrane proteins (OMPs) as a vaccine candidate, we used proteomic mapping and database analysis techniques with extracted OMPs. Also, extracted OMPs were evaluated in several aspects to their safety, immune response in their host and protective effects. Our research has established a proteomic map and database of immunogenic SG-OMPs used as inactive vaccine against salmonellosis in chickens. A total of 22 spots were detected by 2-dimensional gel electrophoresis and immunogenic protein analysis. Eight spots were identified by Matrix-Assisted Laser Desorption/Ionization-Time of Flight-Mass spectrometry (MALDI-TOF-MS) and peptide mass fingerprinting (PMF) and categorized into four different types of proteins. Among these proteins, OmpA is considered to be an immunogenic protein and involved in the hosts' immune system. To estimate the minimum safety dose in chickens, 35 brown layers were immunized with various concentrations of OMPs, respectively. Consequently, all chickens immunized with more than a 50 μg dose were protected against challenges. Moreover, intramuscular administration of OMPs to chickens was more effective compared to subcutaneous administration. These results suggest that the adjuvanted SG-OMP vaccine not only induces both the humoral and cellular immune response in the host but also highly protects the hosts' exposed to virulent SG with 50 μg OMPs extracted by our method.

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