• Korean Journal of Veterinary Service
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• v.13 no.1
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• pp.54-63
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• 1990

We have studied about the safety, immunity and protective potency in swine and experimental am mais of two inactivated vaccine produced with NYJ-1-87 strain of ADV that was isolated in Korea. Result obtained through the experiments were summarized as follows. 1. The safe potency of ADV antigens inactivated with BEI and formaline to mouse & guinea pig was on the whole good, but protective potency rates of those to challenge with ADV were 60-75％ without the differences to two antigens. 2. Safety, immunity & protective potency of ADV antigens inactivated with BEI and formaline to swine were on the whole excellent, except for a mild increase of rectal temperature in some pigs after challenge with ADV. 3. When virus excretion of the experimental groups after challenge with ADV was examed by swabbing of nasal, all pigs of control gorup excreted virus from 2 days p.c., partially to 10 days p.c.. But in vaccinated groups, only 25-50% of all pigs of each group excreted virus during experimental periods. 4. Titers of antibodies in swine & quinea pig vaccinated with inactivated ADV antigens become increased after the 1 weeth p.i. showing the highest liters on the 4-5 weeths p.i.

• Korean Journal of Veterinary Service
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• v.13 no.1
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• pp.32-43
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• 1990

In order to investigate the clinico-pathological and immunohistochemical changes in the rats infected with Aujeszky’s disease virus(ADV), 100 heads of 4 weeks-old rats were inoculated intraperitoneally and intranasally, with the domestically isolated ADV, NYJ-1-87 strain, at $10^{3.0}$ or $10^{5.0}$$TCID_ {50}$/0.2ml. Results obtained through the experiments were summarized as follows : 1. Clinical signs such as dulness, anorexia, pruritus, fascial edema, dyspnea and ataxia were observed from the 2nd day and died at the 3rd to 5th day after ADV inoculation. By necropsy, congestion and hemorrhage were observed in the abdominal organs, while no specific changes were detected in the other organs. 2. In histopathological observation, degeneration and necrosis of the nervous cells, non-suppurative meningoencephalitis, microgliosis and perivascular cuffing were manifested in central nerve system but no specific changes were observed in the other organs. 3. By immunohistochemical staining using peroxidase antiperoxidase, the positive cells were detected in the tissues of kideny, spleen, urinary bladder and lung.

• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.297-307
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• 1990

To establish an agar-gel immunodiffusion (AGID) test for detection of antibodies to Aujeszky's disease virus(ADV) in swine, the precipitating antigens were prepared by four procedures using the Aujeszky's disease virus, NYJ-1-87 strain isolated from the affected piglets in Korea. The optimal condition for AGID test and the properties of the antigens were investigated. To determine the optimal concentration of antigens, four antigens were experimentally prepared by concentrating the viral fluids by 1/30 to 1/200. It was proved that the antigen precipitated with ammonium sulfate at concentration of 1/100 was the most efficient to detect ADV antibodies by AGID test. When the relationship between the concentration of the antigens and the size of precipitating in radial immunodiffusion test was investigated, a high correlation coefficiency at r=0.95 (y=0.23x+23.4) was estimated, In study on the effects of various buffered salt solutions and agars on the sensitivity of AGID test by using the experimental ADV antigens, it was found that 0.05M tris buffer without sodium chloride at pH 7.2 induced the most distinctive precipitating lines, and that there was no significant differences in the sensitivity between the agarose and Noble's special agar. When the efficiency of AGID test was compared with serum neutralization(SN) test, the sensitivity of AGID test was 100% in SN titer over 1 : 16, 91.7% in SN titer of 1 : 8 and 57.1% in SN titer of 1 : 4. The specificity of AGID test compared with the sera with SN titer under 1 : 2 was 98.4%. Protein analysis of the antigens by SDS-PAGE indicated that antigen I and antigen III showed a specific band of polypeptides with molecular weight of 116 K in comparison with the control antigen. Antigen IV, treated with tween-80 and ammonium sulfate, revealed specific polypeptides bands at the molecular weights 45K, 98K and 150 K.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.845-858
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• 1996

The purpose of this study was to establish early diagnostic methods for the detection of Aujeszky's disease viral antigens and nucleic acid in nasal cells, and buffy coats from experimentally infected living pigs by a combination of immunocytochemistry, in situ hybridization with digoxigenin(DIG)-labled probe and electron microscopy. Forty days old piglets were inoculated intranasally with $10^{7.0}TCID_{50}$ of Aujeszky's disease virus (ADV, NYJ-1-87 strain). The viral antigens and nucleic acid of ADV were detected in nasal cells, and buffy coat for 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopical method. The results were compared with conventional methods such as a porcine Aujeszky's disease serodiagnostic(PAD) kit, neutralization test(NT) and virus isolation. 1. The viral antigens, nucleic acids and capsids of ADV were detected in nasal cells, buffy coats from 3 days to 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopy, respectively. 2. When viral antigens were detected by the immunocytochemical technique, a diffuse brown deposit was observed in the nucleus and cytoplasm of nasal cells, buffy coats and PK-15 cells under a microscope. 3. DIG-labeled DNA probe was prepared by amplification of conserved sequence of recombinant ADV-gp50 clone with polymerase chain reacction. When ADV-DNA was detected by ISH with DIG-labeled probe, purplish blue pigmentation were observed in the nuclei and cytoplasms of ADV-infected cells under a microscope. Positive signals were observed in nasal cells and in the buffy coat and PK-15 cells at the first day after inoculation. 4. Where ADV-capsids were detected by transmission electron microscopical method, aggregation of capsids was observed in the nuclei and cytoplasms of nasal cells, buffy coats and PK-15 cells. The results suggested that these methods were considered as the highly sensitive and reliable tools for rapid and confirmative diagnosis of Aujeszky's disease in living pigs.

• Korean Journal of Veterinary Research
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• v.30 no.2
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• pp.177-186
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• 1990

To investigate the etiology, pathogenicity and virological properties of NYJ-1-87 strain of Aujeszky's disease virus (ADV) that was isolated from the diseased piglet in Korea, the virus at $10^{6.0}TCID_{50}/0.1ml$ was inoculated intranasally and subcutaneously into 30 to 35 days-old piglets. Results obtained through the experiments were summarized as follows. 1. Ten of the infected piglets were clinically observed for 15 days. On the 2nd day post-inoculation(pi), the signs of pyrexia, anorexia and convulsion were noted. On the 4th to 7th days pi, nervous signs of incoordination and intermittent spasm were shown in the most of piglets, and one out of 5 piglets infected intranasally was died with severe nervous signs at the 7th day pi. The signs became relieved on the 8th day pi and all of remainder were completely recovered on the 13th to 14th days pi. 2. In hematological study, prominent decrease in the number of total leukocyte and lymphocyte was shown in the ADV-infected piglets on the 6th day pi. On the 8th day pi, the cell numbers were slightly increased and returned to normal level on the 10th day pi. 3. Viral excretion of the ADV-inoculated piglets was examined by swabbing of nasal and oral cavities, and rectal feces. During the periods of the 3rd to 11th days pi, the virus was excreted intermittently from nasal and oral cavities, and rectal feces. The nasal excretions were shown the highest virus concentration of $10^{5.2}TCID_{50}/0.1ml$ at the 5th day pi. 4. Recovery of the inoculated virus from various organs of the piglets that were died or experimentally slaughtered was attempted, and the virus was isolated from the tissues of brain and tonsil by the cultured cell-inoculation method. The highest recovery rate was noted in the tonsil. By indirect immunofluorescence antibody assay using ADV-monoclonal antibody, the viral antigens were detected in tissues of spleen and liver as well as brain and tonsil on the 7th to 9th days pi. The virus was not isolated from blood and the tissues of lung and kidney throughout the experiments. 5. Titers of virus neutralizing antibody in the piglets experimentally infected with ADV became increased after the 6th to 9th days pi in both of intranasal and subcutaneous inoculation showing the highest titers of 64 to 128 on the 29th day pi. When the antibody levels were measured by radial immunodiffusion enzyme assay, the reactive diameter was enlarged to be positive after the 4th to 6th days pi in both of intranasal and subcutaneous inoculation showing the largest diameter of 13 to 14mm on the 29th day pi.