• Korean Journal of Veterinary Research
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• v.35 no.4
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• pp.753-758
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• 1995

The study was carried out to investigate the pathogenicity of EHV isolate to hamsters and mice and immunogenicity of experimentally produced. vaccine were evaluated in the horses. Hamsters infected. intranasally with $LC_1$ isolate showed symptoms of nasal discharge, conjunctivitis and body weight loss during the observation period of 12 days after infection, while only slight depression and body weight loss were noticed with mice infected with $LC_1$ indicating that hamsters are more susceptible to the virus. Antibody titer of mice and hamsters were gradually increased to highest level of 1:2560~10240, 1:640~1280, respectively, at 7~12 days post vaccination. Horses immunized against $LC_1$ killed vaccine reached to maximum antibody titer of 1:20480 around 4 weeks after 1st vaccination and declined after 12 weeks post vaccination. No significant antibody increase were detected after 2nd vaccination. Mean body temperature and mean total leukocyte counts remained within normal range and no adverse reaction were noticed after vaccination.

• Korean Journal of Veterinary Service
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• v.22 no.1
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• pp.37-42
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• 1999

This study was conducted to determine whether the vaccination programs for the control of Newcastle disease(ND) would affect the immune status of Ogol chicken in Yeonsan, Chungnam province. The results obstained are summerized as followes ; 1. Maternal antibody titers level of Ogol chickes was 2 $log_2$. 2. The efficacy of the Newcastle spray and drinking water vaccine was evaluated by the antibody responses and the protection rates(over 90% ) after challenge with a virulent Newcastle disease vims at 14 days. 3. Spray vaccine caused on visible side reaction like respiratory symptoms, but it is declined of the 4~5 days.

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.82-88
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• 1999

In this study, to evaluate newly developed Japanese encephalitis (JE) vaccine candidate CJ50003, we assessed its immunogenicity along with a previously commercialized inactivated JE Biken vaccine. The CR0003 viral antigens produced in Vero cells were administered suhcutaneouly to mice either with alum-adjuvanled or free form. The ELISA titers and neutralizing (NEUV antibody titers accounting for major protective immunity in JE were determined. Mice given alum-adjuvanted vaccine had a 10 times higher antigen-specific NEUT antibody response than did those which {lad received free antigens. This NEUT antibody response was maintained until day 168 with NEUT titer more than 1:160. Even with the 0.5 ng of alum-adjuvanted antigen dose, NEUT titer was induced more than 1:10 which is considered as an evidence for seroconversion and protection. Thc mice immune sera had a similar rate of cross-reactivity against three different viral antigens, Nakayama-NlH, P3 and SA14; as determined by ELISA assay. In a mice challenge model, vaccination with the GI50003 conferred more protection than with commercialized Biken vaccine against Nakayama virus. These data demonstrated that CJ50003 vaccine candidate has an excellent prophylactic efficacy and implicated it has a strong potential for further development and commercialization.

• Korean Journal of Poultry Science
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• v.17 no.3
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• pp.225-232
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• 1990

The immune responses of commercial layer chickens against Newcastle disease (ND) were compared among different administration methods and times of vaccination during 4 weeks of age. A total of 372 day-old chickens were devided into 4 groups of 93 birds each. Each of 3 groups was received a commercially available B$_1$ live vaccine via drinking water, eye instillation or spray method at one, 14 and 28 days of age. One group was used as an unvaccinated control. At two and 4 weeks after each time of vaccination, 15 birds from each group were challenged with virulent ND virus at the dose of 10$^{5}$ EID$_{50}$ per bird to examine the pretection rate. Ten to 15 birds from each group were bled at two weeks intervals from day old to 8 weeks of age to determine hemagglutination inhibition antibody titer. The protection rate was generally low regardless of the times of vaccination although two or more times vaccination gave higher protection than once vaccination. The low protection was considered due to low titer of the vaccine used since the vaccine titer was less than 10$^{2.5}$ EID$_{50}$ per bird. Spray method gave better protection compared to eye instillation of drinking water method which resulted in lowest response. When birds were challenged majority showed clinical signs on ND between 3 and 6 days after challenge. Death occured one or two days after onset of symptoms. Major clinical signs observed were depression (96%), drowsy(90%), anorexia (84%), diarrhoea (29%), difficult breath (15%) and torticollis (10%). Hemorrhagic lesions on post mortem were seen in duodenum (51%), trachea(36%), illeum (13%), ceacal tonsil (11%), proventriculus (10%) and some other organs. When birds were challenged majority showed clinical signs on ND between 3 and 6 days after challenge. Death occured one or two days after onset of symptoms. Major clinical signs observed were depression (96%), drowsy(90%), anorexia (84%), diarrhoea (29%), difficult breath (15%) and torticollis (10%). Hemorrhagic lesions on post mortem were seen in duodenum (51%), trachea(35%), illeum (13%), ceacal tonsil (11%), proventriculus (10%) and some other organs.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.738-742
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• 1999

The objective of this experiment was to investigate the effect of Newcastle disease virus (NDV) and infectious bursal disease virus (IBDV) vaccination on performance of broiler chicks for five weeks. Two types of poultry houses and three patterns of vaccination ($NDV^-/IBDV^-$, $NDV^+/IBDV^-$ and $NDV^+/IBDV^+$) were factorially assigned to six treatments. NDV, B1 strain and IBDV, Bursin-2 vaccine were orally administered at 5, 14 and 7, 18 days, respectively. Forty eight hundred chicks were grouped into four replications with two hundnyd hybro $\times$ hybro chicks per each treatment. Weight gain, feed conversion ratio (FCR), mortality and product index were surveyed at the end of experiment. Bursa index and IBDV antibody titer of chicks were weekly measured. Weight gain of chicks vaccinated with $NDV^+/IBDV^+$ was significantly increased compared to that of other treatments at both window and windowless poultry houses (p<0.05). Chicks vaccinated with $NDV^+/IBDV^+$ also showed significantly improving the FCR and mortality compared to those of other treatments at both poultry houses (p<0.05). The bursa indecies of both poultry houses were high from one-day- to three-weeks-old, but were low for the rest of two weeks. IBDV antibody of all chicks was detected 100% by agar gel precipitation (AGP) test at one day old, but was not detected in $NDV^-/IBDV^-$ and $NDV^+/IBDV^-$ treatments at four weeks old. However, it showed 100% in $NDV^+/IBDV^+$ treatment. Antibody titer using ELISA showed similar trend to that of AGP test. The results of this experiment confirmed that IBDV and NDV combined vaccine significantly improved the performance of broiler chicks.

• Korean Journal of Poultry Science
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• v.30 no.3
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• pp.197-202
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• 2003

Feeding probiotics in broiler chicks still critical in several aspects. Thus, this study was conducted to investigate the impact of feeding multiple probiotics on performance, intestinal microflora, blood cholesterol and ND antibody vaccine titer in broiler chicks. Three hundred twenty one day old male broiler chicks(cobb ${\times}$ cobb) were divided into four levels of multiple probiotics(0, 0.1, 0.2, 0.3%) with five replicates for 35 days. Basal diets contained 21.5, 19.0% CP and 3,100 kcal/kg ME for starting and finishing period, respectively. Weight gain, feed intake and feed conversion were measured weekly. The number of Salmonella, E. coli, Lactobacillus, and yeast were examined from ileum and cecum at the end of experiment. ND vaccine titer, cholesterol were detected from sera. Weight gain of birds fed probiotics were 669.33, 679.75 at the level of 0.1 and 0.2% supplemental groups for starting period. It was also improved in those treatments for finishing period and higher than control for total period. Feed conversion tended to be improved compared to that of control by the supplementation of probiotics for the first three weeks and seemed to show the similar tendency for the rest of two weeks. It was 1.611, 1.621 for the entire feeding period and improved compared with control. Total salmonella, was not decreased in ileal digesta of birds fed the probiotics compared with control, whereas the number of yeast increased in 0.1% treatment. However, the number of Lactobacillus and yeast in cecum was higher than control. Even though the blood cholesterol seem to high in 0.1% probiotics treatment, the ratio of HDL to total cholesterol showed higher than control. ND vaccine titer of birds fed probiotics were significantly higher than control (P<0.05). These results 0.1% multiple probiotics would be possible to improve the performance of broiler chicks and ND vaccine titer.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.144-150
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• 1997

We developed a simple and efficient method to prepare a Pseudomonas vaccine of outer membrane (OM) proteins free from lipopolysaccharide (LPS). A three step purification process including extraction, ultrafiltration and ultracentrifugation effectively removed LPS from the OM protein fraction. Approximately 2 mg of the OM proteins was obtained from 1 g of wet cell. LPS contaminant in the vaccine preparation was less than 0.003% (w/w) of protein and protease activity was not detectable. To achieve a wide range of protection, OM proteins prepared from four attenuated P. aeruginosa strains were mixed in equal amounts and used as a vaccine, which elicited in rabbits a high titer of antibody reactive to all of the seven Fisher types. The antisera from the immunized rabbit had a strong reactivity to vaccine proteins larger than 25 kDa. In a burned mouse infection model, immunization with the vaccine significantly enhanced bacterial clearance in the Pseudomonas infected skin. The vaccination also provided mice an excellent protection against Pseudomonas infection (11, 16). Data on antigenicity, mutagenicity, acute, subacute toxicity and pharmacological tests confirmed the safety of the vaccine (1, 3, 10, 12, 17). These data demonstrate that this method can be applied to manufacture a bacterial vaccine of OM proteins with safety and prophylactic efficacy at a practical low cost.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.489-499
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• 2019

Subunit vaccines are safer and more stable than live vaccines although they have the disadvantage of eliciting poor immune response. To develop a subunit vaccine, an effective delivery system targeting the key elements of the protective immune response is a prerequisite. In this study, oxidized carbon nanospheres (OCNs) were used as a subunit vaccine delivery system and tuberculosis (TB) was chosen as a model disease. TB is among the deadliest infectious diseases worldwide and an effective vaccine is urgently needed. The ability of OCNs to deliver recombinant Mycobacterium tuberculosis (Mtb) proteins, Ag85B and HspX, into bone marrow derived macrophages (BMDMs) and dendritic cells (BMDCs) was investigated. For immunization, OCNs were mixed with the two TB antigens as well as the adjuvant monophosphoryl lipid A (MPL). The protective efficacy was analyzed in vaccinated mice by aerosol Mtb challenge with a virulent strain of Mtb and the bacterial burdens were measured. The results showed that OCNs are highly effective in delivering Mtb proteins into the cytosol of BMDMs and BMDCs. Upon immunization, this vaccine formula induced robust Th1 immune response characterized by cytokine profiles from restimulated splenocytes and specific antibody titer. More importantly, enhanced cytotoxic $CD8^+$ T cell activation was observed. However, it did not reduce the bacteria burden in the lung and spleen from the aerosol Mtb challenge. Taken together, OCNs are highly effective in delivering subunit protein vaccine and induce robust Th1 and $CD8^+$ T cell response. This vaccine delivery system is suitable for application in settings where cell-mediated immune response is needed.

• Korean Journal of Veterinary Service
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• v.46 no.3
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• pp.193-202
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• 2023

Porcine epidemic diarrhea is an infectious intestinal disease caused by the porcine epidemic diarrhea virus (PEDV). Especially, when suckling piglets are infected, the mortality rate is close to 100%. PEDV is classified into G1 and G2 types based on genetic differences. The G2 type PEDV outbreak in the United States in 2013 was highly pathogenic and contagious, and it has spread worldwide and caused continuous economic losses. Most commercial vaccines used are G1 type vaccines, and existing vaccines do not fully protect piglets due to genetic differences. In this study, we evaluated the safety of the newly developed G2 type attenuated HSGP vaccine strain by inoculating it into piglets and testing whether the vaccine virus spreads to the non-vaccinated, negative pigs and whether the vaccine reverts to its virulence during serial passage experiments. Each experiment lasted for 7 days for each passage, and fecal viral titers, clinical symptoms, and weight gain were measured daily. After the experiment, necropsy was performed to measure intestinal virus titer and pathological evaluation. As a result of the first passage, no transmission of the vaccine virus to negative pigs co-housed with vaccinated pigs was observed. In addition, after four consecutive passage experiments, the clinical symptoms and small intestine lesions were gradually alleviated, and no virus was detected in the feces in the fourth passage experiment. Therefore, it was concluded that the vaccine was safe without virulence reversion in accordance with the guidelines of the current licensing authority. However, further studies are needed on the genetic changes and biological characteristics of the mutant virus that occur during successive passages of the attenuated vaccine since the replication and clinical symptoms of the virus increased until the third passage during successive passages of the vaccine virus. Based on this study, it was concluded that virulence reversion and safety evaluation of attenuated vaccines through serial passage in target animals can be useful to evaluate the safety of attenuated viruses.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.110-116
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• 2000

Duck viral hepatitis is an acutic, highly infectious viral disease of young ducklings. The most practical means for controlling duck viral hepatitis is the vaccination of ducklings or of a breeding stock. We attempted to develop a vaccine strain of duck hepatitis virus (DHV) using a Korean isolate by serial chicken embryo passages. The propagation of DHV in chicken embryos was carried 140 passages. After the $50^{th}$ passage, of which the virus was non-pathogenic for ducklings, approximately every $20^{th}$ passage of the virus was tested for vaccinal efficacy. Both the $70^{th}$ and $90^{th}$ passage of the virus gave good protection against challenge infection to a DHV-DRL reference strain(type 1) and a virulent Korean isolate. The $110^{th}$, $125^{th}$ and $140^{th}$ passage of the virus were less protective than the $70^{th}$ and $90^{th}$ passage, which means that more than $110^{th}$ passage may lead to over-attenuation of the virus. Ducklings vaccinated with the chicken-embryo-adapted virus by oral, intramuscular or eye drop administration showed earlier resistance to challenge infection from 3 to 7 days postvaccination. Of the above methods, ducklings vaccinated intramuscularly presented the most rapid resistance against challenge. The minimum immune dose of the chicken-embryo-adapted virus in ducklings was also studied. Ducklings inoculated with a dose of $10^{2.0}\;ELD_{50}$ and below were not fully protected against challenge with a virulent DHV, showing a protection rate of 67% to 73%, but ducklings inoculated with a dose of $10^{3.0}\;ELD_{50}$ and over were completely protected. The virus yield of the chicken-embryo-adapted DHV was examined at 24hrs and 48hrs of the incubation time in the allantoic fluid, embryo head and embryo minus head of the embryonating egg. In all three components, the titer of the virus was higher at 48 hours than that at 24 hours after incubation. And the titer of the virus was higher in the embryo minus head, embryo head and the allantoic fluid, in order. Field trials for evaluating the efficacy of the attenuated DHV as a live vaccine were done in duck farms with about 25% mortality of flocks resulting from duck viral hepatitis. After the use of the experimental vaccine, the mortality due to duck viral hepatitis was dramatically reduced in the farms. These results indicated that the attenuated DHV using a Korean isolate could be a good candidate as a live vaccine strain of DHV in Korea.