• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1271-1279
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• 2009

To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

• Journal of Veterinary Clinics
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• v.26 no.2
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• pp.134-137
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• 2009

Although there is circumstantial evidence that infectious bronchitis(IB) in the Korean layer industry has contributed to severe economic losses, the seroprevalence against IB virus(IBV) and risk factors associated with seropositivity are not well known. During May to October 2007, 820 blood samples were randomly collected from 41 laying hen flocks(20 birds in each flock) with $\geq$ 3,000 birds of 18 week of age or older in three provinces of Korea. The samples size was determined considering a flock-size range of 3,000-65,000 birds, an expected bird-level seroprevalence of $\geq$ 15%, and a 95% level of confidence. Serum samples were examined using a hemagglutination inhibition test for antibodies to IBV. The overall apparent flock-level seroprevalence was 46.3%(95% CI, 31.1-66.6) with no statistically significant differences among provinces(X=1.205, p>0.05). There were 19 positive flocks with one to eight seropositive birds, and 11 of these had one or two seropositive birds. None of the measured parameters were significantly associated with seropositivity against IBV in a subsequent multivariable logistic regression analysis. A longitudinal risk factor studies considering management and vaccination characteristics possibly associated with the IBV flock prevalence would be beneficial.

• Journal of Practical Agriculture & Fisheries Research
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• v.20 no.1
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• pp.89-96
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• 2018

The object of this study was to evaluate the change of antibody titers on virus strains after inoculation with commercial killed equine influenza (EI) vaccines in horses. Serum antibodies of 20 Thoroughbred yearlings were detected using hemagglutination inhibition test for 41 weeks. Second vaccination is inoculated 4 weeks after the initial vaccination. Most of antibody titers were not increased until 4 weeks after first vaccination. The highest titers were detected 6-10 weeks after vaccination. The titers were decreased slowly and maintained for 16 weeks after inoculation. We could barely detect the antibody 41 weeks after vaccination in most cases. Vaccine anergia were appeared in 3 horses (15%) but it depended on virus strains. A/Equine/La Plata/93(H3N8) strain that induce high and durable antibody responses was the most effective among three strains. This study presents the first comprehensive data on the endurance of antibody titers against EI. Our data also suggests that yearlings should be inoculated three times in order to maintaining optimal antibody titers against EI. We speculate the causes of anergia were vaccine break down or individual specificity. Further research is needed to investigate immunological unresponsiveness. This was the first study on strain of equine vaccine in Korea.

• Korean Journal of Poultry Science
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• v.18 no.3
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• pp.183-196
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• 1991

In order to establish ELISA method to detect antibody against IBV various factors involved were examined. Antigen was prepared from Massachusetts type IBV which is known to be one of serotypes distributed most widely. The virus was grown in embryonated SPF chicken eggs. Allantoic fluid harvested was processed to ultracentrifugation and sucrose density gradient centrifugation to produce a purified antigen The antisera selected from the field samples based on hemagglutination inhibition test were used as the standard positive and negative sera for this study and the results obtained were summarized as follows. 1 , It was found that ELISA test was satisfactory when the purified antigen was coated on the plate in the amount of about 40ng protein per well. In case of the phospholipase treated hemagglutinating antigen it gave satisfactory results when the each well wns coated with 1.2 to 2.5 hemagglutinating unit which was equivalent to 40 to 90ng of protein. 2. There was no significant difference in the ratio of optical density of positive to that of negative serum whether the coated antigen was held for 1 hour at 37$^{\circ}C$ or it was held overnight at 4$^{\circ}C$. The coated antigen could be kept in dried state without change of antigenecity for at least one month of experimental period at 4$^{\circ}C$. 3. There was a big variation in the optical density and P/N values depending on the maker of the plates and on the plate of the same maker. 4. It was found that background optical density was negligible when serum was diluted more than 1：50 and serum dilution of 1：100 appeared to be appropriate as a routine test dilution to screen the antibody. 5. Optical density was fairly constant 15 minutes afterward from the time substrate was treated and during the 4 hours after stopper was treated. 6. There was a low correlation(r＝0.42) between ELISA and HI test. However, when 74serum samples were tested for the IBV antibody, 98.7% were found to be positive by both tests in which titers of 2$^{6}$ or more by HI test and P/N values of 1.4 or more by ELISA were considered to be positive, 7 Day-old IBV vaccinated chickens shows a similar antibody decay and rising pattern until 8 weeks of age by the two tests, ELISA and HI.

• Korean Journal of Veterinary Service
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• v.28 no.2
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• pp.121-146
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• 2005

Newcastle disease (ND), infectious bronchitis (IB), low pathogenic avian Influenza (LPAI) and fowl typhoid (FT) have been known as egg drop laying diseases because of the serious layer damage from mass zone layer. In this study, such egg drop laying diseases were investigated. To access this study, we peformed to evaluate antibody titers in serum and isolated bacteria and virus from organs and feces on May, July and September in 2003. The distribution of ND from January to May, IB and LPAI from October to February of the next year, and FT from March to September were inspected by the question survey in 21 farms. ND revealed to be positive rates of 490 to 474 $(96.7\%)$ in May, 510 to 506 $(99.2\%)$ in July and 510 to 510 $(100\%)$ in September with hemagglutination inhibition (HI) test. The mean antibody titers were 10.2, 9.9 and 10.2, respectively. With regard to IB, 484 out of 490 samples $(98.7\%)$ in May, 508 of 510 $(99.6\%)$ in July and 509 of 510 $(99.8\%)$ in September showed positive results and the mean antibody titers were gradually increased with 8.2, 9.0 and 9.4, respectively. According to HI test of LPAI, the positive results were shown in 442 of 480 $(92.1\%)$, 394 of 494 $(79.8\%)$ and 402 of 483 $(83.2\%)$ in May, July and September, respectively The mean antibody titers were decreased with 4.6, 4.3 and 4.0. The distribution of LPAI also elicited the positive rates of 480 to 475 $(99.0\%)$ in May, 494 to 485$(98.2\%)$ in July, 483 to 472 $(97.7\%)$ in September as determined by ELISA and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. Compared ELISA results with HI test of LPAI the positive results were 480 to 422 $(92.1\%),\;475(99.0\%),\;494\;to\;394 (79.8\%),\;485 (98.2\%)\;and\;483\;to\;402(83.2\%),\;472(97.7\%)$. Therefore, the positive rate determined by ELISA was higher than that of HI test with 6.9, 18.4 and $14.5\%$, respectively. When performed RT-PCR for ND using organ and feces samples, the pathotypes were detected $5(15.6\%)\;in\;May,\; 2(5.3\%) in\;July,\;2(7.1\%)$ in September but there is no samples showing positive band for LPAI. In attempt to isolate Salmonella gallinarum, bacteria were obtained from 4 cases (12.5%) in May, 9 (23.6%) in July, 5 (17.8%) in September. Thus the highest rate for isolation revealed to be shown in July When evaluated the antimicrobial susceptibility to 18 isolated strains of 5. gallinarum, bacteria were sensitive to trimethoprim/sulfamethox$(61.1\%),\;kanamycin\;(55.5\%),\;ampicillin\;(55.5\%)$ and amoxacillin/clavulanic acid $(55.5\%)$, cephalothin $(50.0\%)$, but resistant to penicillin $(88.9\%)$, streptomycin $(88.9\%)$, erythromycin $(83_4\%)$ and tetracycline $(61.1\%)$. According to HI test of ND and LPAI using captured 164 wild Korean tree sparrows (Passer nontanus), the positive rates were $47.6\%\;and\;57.3\%$, and the mean HI titers were 5.32 and 4.02, respectively. 71 $(43.2\%)\;and\;58(35.3\%)$ in captured sparrows also showed more than 4 titers for HI test to ND and LPAI, respectively However, the attempt for isolation of viruses failed in all samples.

• The Journal of the Korean Society for Microbiology
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• v.15 no.1
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• pp.77-84
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• 1980

The purpose of this paper is to study the incidence of humoral antibody to reovirus type 2 in the sera of the Koreans and animal(swine) at random. All the 614 of human beings and 877 of swine sera were collected during the period from June to December, 1979, from the healthy persons in the National Seoul Hospital and swine blood was collected from 25 different areas of June to 30th of September in 1977. The HI test was put with plastic plates according to the methods by Rosen(1960 a and 1974). The total 73.29% of the 614 cases in human and the 61.80% of the 877 in swine confirmed as a hemagglutination inhibition antibodies. In human the 76.47% of the 442 male cases and the 65.12% of the 172 female ones were confirmed to have humoral antibodies. The positive rate was widely shown in each age group. But the 31 to 50 old age groups showed a little higher than any other age group, which came to the 85.71% in 41-50 and the 78.72% in 31-40 old age groups. The monthly distribution of HI antibody was shown to reach the 93.22% of the 59 cases in June. This per cent was much higher than of any other distribution. Accordingly, the auther came to the conclusion that there is reovirus type 2 in all the parts of Korea and most of the Koreans have the higher rates of antibody. However, the positive rate of antibody was the 542 out of the 877 cases(61.8%) from the swine and antibodies was confirmed from the 25 different areas in Korea. The seasonal distribution of the antibody showed these high rates. In domestics animals; blood from the swine showed that distribution of HI antibodies to reoviras type 2. These antibody appears from the various areas of the province in Korea. For this reasons, reovirus was widely distributed in this country.

• Pediatric Infection and Vaccine
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• v.5 no.2
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• pp.258-266
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• 1998

Purpose : Cases of adenoviral penumonia with rapidly progressive clinical course were experienced. We reviewed these patients in viewpoint of clinical manifestation and adenoviral serotypes. Methods : Culture and indirect immunofluorescence for respiratory viruses including respiratory syncytial virus, influenza virus, parainfluenza virus, adenovirus was done with nasopharyngeal aspirates from patients who admitted due to respiratory infections in Fatima Hospital, Masan from Nov. 1996 to Jul. 1997. Cultured adenovirus was serotyped by both neutralization and hemagglutination inhibition test. Medical records were reviewed for 5 patients with respiratory failure from adenovirus was isolated and serotyped. Results : The total number of examined patients was 460 patients. We isolated respiratory viruses in 143(30.9%) patients. Adenovirus was isolated from 66 out of 143(46.2%) patients. During Jan 1997 to May 1997, five patients with ages of 18 days to 11 months who were infected by adenovirus and had high fever with dyspnea and required assisted mechanical ventilation. One patients discharged against doctor's advice then died. Two of four patients had complications of disseminated intravascular coagulation; two had bronchiolitis obliterans. Two isolates were serotype 7, and one was serotype 5, and two were untyped. Conclusion : Severe pneumonia caused by serotype 7 continued to occur in 1997 following the epidemic in 1996, and severe pneumonia may also be caused by serotype 5 and other serotypes.

• Korean Journal of Poultry Science
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• v.39 no.2
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• pp.97-104
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• 2012

Newcastle disease virus (NDV) infects a variety of birds with a wide range of clinical signs from asymptomatic to severe. During a 10-month period in 2011, a total of 1,024 sera from wild birds including 42 species of birds in 8 orders were collected and the seroprevalence of NDV in wild birds was evaluated by hemagglutination inhibition (HI) test. Evidence of NDV infection was observed in 12.6% (129/1,024) of wild birds with a maximum prevalence reported in Mandarin duck (27.8%, 32/115) followed by Mallard duck (20.8%, 57/274), Spot-billed duck (11.9%, 36/303), Pintail (2.9%, 1/34), Black-tailed gull (2.9%, 1/34), White-fronted goose (1.8%, 1/56) and Common teal (1.4%, 1/69). None of the other 35 species of wild birds were antibody-positive for NDV. Mandarin duck, Mallard duck and Spot-billed duck showed high sero-prevalance of 12.2% to 42% during winter season (November to March). Our results indicate that Mandarin duck, Mallard duck and Spot-billed duck might be natural reservoirs for NDV in Korea and the prevalence of NDV infection in wild birds displayed a seasonal pattern with high prevalence of NDV in winter season (November to March).