• 한국동물위생학회지
• /
• 제14권2호
• /
• pp.134-142
• /
• 1991

We surveyed the antibody titers in the slaughtered pigs by enzyme immunodiffusion method for the epidemiological distribution of Aujeszky's disease in Korea. And also we observed the clinical symptoms, histopathological findings and antibody titers by serologic neutralization test in experimental animals infected with Aujeszky's disease virus(ADV) isolated from Korea. The results of the experiments were summarized as follows. 1. We detected 2 pigs infected with ADV of 1000 in 1990 and 1 of 600 in 1991 by enzyme immunodiffusion method. 2. In histopathological findings of experimental animals inoculated ADV, the typical characteristics of Aujeszky's disease were not observed in pig, but edema and degeneration in the epidermis were observed in rats, vasculoendotheriosis, perivascular cuffing and cellular infiltration in the cerebrum were appeared in rabbits and perivascular cuffing and forcal infiltration of glial cells in the cerebrum were appeared in mice. 3. The increasing antibody titers(1 : 64) in the serological neutralization test were ascertained in 2 carrier pigs inoculated ADV. 4. Rabbits, mice and rats died all about 5 days after inoculation, but pigs and chickens didn't die. 5. In 1 cat and 2 rabbits inoculated with the Aujeszky's disease virus(ADV), the typical clinical findings of Aujeszky's disease were observed in rabbit, but not in cat and so we slaughtered without any signs of Aujeszky's disease.

• 한국동물위생학회지
• /
• 제19권1호
• /
• pp.1-29
• /
• 1996

This study was conducted to elucidate the pathogenesis of Aujeszky's disease virus(ADV) by histopathologic examination. The first Korean ADV Isolate, which was isolated from piglets with clinical signs of Aujeszky's disease in Yangsan(YS) county, Kyungnam province, was inoculated into 32 days old piglets with a dose of $10^{5.9}$$TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at intervals of every 24hrs for 8 days. The virulence of YS strain was determined by the observation of clinical signs, gross findings, and histopathologic changes in tissues. The virus recovery test was performed from brain, spleen, lung and tonsil in cell culture. The pathogenesis of YS strain was determined by the observation of histopathologlc lesions in CNS and neuronal tracts. The major clinical signs were fever, anorexia, dyspnea, constipation, tremor, ataxia, circling movement, hindleg paralysis and salivation. The clinical signs were more severe in piglets of the group inoculated intranasally than those of the intramuscularly inoculated gorup. Lymphocytopenia was detected on day 5 to day 6 postinoculation (PI). The ADV was recovered from the tissue homogenates of tonsil, lung, spleen and cerebrum in cell culture. The highest virus titer was detected from tonsil between day 6 and day 7 PI. Reddish sublobar consolidation foci were scattered in the apical and cardiac lobes of lung. Although yellowish necrotic foci were detected in tonsil and liver, hemorrhagic lesions were mainly observed in heart, kidney and lymph nodes. Histopathologically, degeneration and necrosis of nerve cells, nonsuppurative meningoe-ncephalitis, nodular gliosis and perivascular cuffings were observed in CNS. Multifocal fibronecrotic foci were observed in lung, liver, lymph nodes and spleen. The major pathologic changes were detected in the midbrain, pons and medulla oblongata. Eosinophilic intranuclear inclusion bodies were mainly observed in epithelia and /or macrophages of tonsil, liver, lung, spleen and submandibular lymph nodes, and neurons of brain, respectively. Observation of viral particles at various stages of replication were possible from the endothelial cells of the alveolar capillaries and tonsillar crypt epithelia by transmission electron microscope.

• 한국동물위생학회지
• /
• 제13권1호
• /
• pp.32-43
• /
• 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.

• 대한수의학회지
• /
• 제34권3호
• /
• pp.529-536
• /
• 1994

To establish more specific and simple diagnostic methods for detection of the antibodies and antigens of Aujeszky's disease virus(ADV), we designed indirect dot-immunoassay(IDI) and double sandwich dotimmunoassay(DSDI) using the solid phases of nitrocellolose paper and polystyrene plate. The diagnostic efficacy of these methods was investigated. As the sensitivity of IDI was tested by various virus concentration, the specimens with the virus titer above $10^{4.0}TCID_{50}/0.2ml$ showed positive reaction, but that below $10^{1.0}TCID_{50}/ml$ revealed negative. Tonsil emulsion at the virus titer of $10^{4.5}TCID_{50}/0.2ml$ showed the highest sensitivity as diluted by 1/100. In detection of ADV antigens from the various tissues of the rats and pigs infected with ADV, IDI using monoclonal antibody showed the higher specificity as compared with IDI using polyclonal antibody and virus isolation method. The efficacy of the DSDI for detection of ADV antibody was compared with other tests. The sensitivity of DSDI was higher than virus neutralization(VN) and agar gel immunodiffusion test(AGID). Meanwhile, specificity of DSDI was lower than AGID, but similar to IDEA. In comparison with VN test, DSDI showed 96.9% agreement to VN test that is the highest of three tests. In general, application of polyclonal antibody in both tests caused the higher sensitivity but the lower specificty.

• 대한수의학회지
• /
• 제30권3호
• /
• pp.297-307
• /
• 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.

• 대한수의학회지
• /
• 제36권4호
• /
• pp.859-871
• /
• 1996

This study was conducted to elucidate the distribution of Aujeszky's disease viral nucleic acids and antigens in the central nervous system (CNS) of piglets. The first Korean isolate of Aujeszky's disease virus(ADV) that isolated from naturally infected piglets in Yang San, was inoculated into 32 day old piglets with $10^{5.9}TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at every 24hrs for 8 days. The immunohistochemistry (IHC) was conducted to detect the viral antigens in paraffin-embedded tissue sections using avidin-biotin-peroxidase complex (ABC) method. The viral nucleic acids were detected by in situ hybridization (ISH) using ADV specific DNA probe labeled with digoxigenin. The ADV antigens were detected in reticuloendothelial cells of spleen, lymph nodes and tonsil, alveolar walls, leptomeningeal vascular walls, inflammatory foci of each organ, and nerve cells. The viral nucleic acids were detected in the spinal trigeminal nucleus and its tracts of the pons and medulla oblongata by the ISH technique. The pathways of AD viruses in CNS were determined by IHC and ISH. In the intranasally inoculated group, the viruses in nasal mucosa moved to medulla oblongata and pons through the trigeminal nerve. In case of intramuscullarly inoculated group, viruses moved to brain via lymphoid organs or spinal nerves from sciatic nerves.

• 대한수의학회지
• /
• 제33권2호
• /
• pp.241-247
• /
• 1993

Forty day old piglets were intranasally inoculated with 2ml of Aujeszky's disease virus (NYJ-1-87 strain, $10^{7.0}$ $TCID_{50/0.2}ml$), and the viral antigens were detected in nasal and circulating white blood cells for 20 days after inoculation by immunocytochemical method. Antibody titers in the blood were also detected by neutralizing test and Aujeszky's disease serodiagnostic kit(Choong Ang) in this periods. 1. Viral antigens were detected by the immunocytochemical technigue, and positive reactions were observated in nasal cells from the 2nd to the l0th days after inoculation and circulated white blood cells from the 4th to the 12th days after inoculation. 2. In neutralization test antibodies levels showed titers of 2 on the 8th day, 8 on the l5th day, 16 on the 18th day and 32 on the 20th day after inoculation. In serodiagnostic kit test positive reactions were observed after the 15th day after inoculation.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제35권2호
• /
• pp.118-122
• /
• 2017

Porcine pseudorabies virus (PRV) causes the Aujeszky's disease (AD) which is economically important disease in the swine industry worldwide. Killed or live vaccines have been used to control this disease, but their efficacy and side effects remain problems to be solved. To solve these problems, in this study, production of recombinant PRV glycoprotein gB, gC and gD was investigated in insect expression system. Glycoprotein gB, gC and gD are regarded as the major immunogenic antigens in PRV. Abundant production and immunogenicity of glycoprotein gB, gC and gD were confirmed by SDS-PAGE and Western blot analysis, respectively. Optimal infection dose and time were also determined for the production of each recombinant PRV glycoprotein. Confirmation of glycosylation of recombinant gB, gC and gD suggested their usefulness as antigens for the development of diagnosis kit or vaccines for Aujeszky's disease.

• 대한수의학회지
• /
• 제34권2호
• /
• pp.327-333
• /
• 1994

The purpose of this study was to establish a rapid diagnostic method detecting Aujeszky's disease virus (ADV) DNA in the cultured cell monolayers (PK-15) and tissue sections of ADV(NYJ-1-87)-infected rats and pigs by in situ hybridization(ISH). Detection of specific ADV-DNA in infected cells was conducted by radiolabeled ISH method using $^{32}P-labeled $ DNA probe (BamH1 7 fragment) which contains a 6.3 Kb ADV-DNA insert. Where ADV-DNA was detected by radiolabeled ISH, the deposition of black photographic grains occurred in the nuclei and the cytoplasms of ADV-infected cells. Positive hybridization signal was often observed in the spinal trigerminal nucleus of the pons, the nucleus of the trigerminal ganglion neuron and the epithelial cells of tonsillar crypts. The results suggested that ISH is considered as a highly sensitive and reliable tool for confirmative diagnosis of this viral disease.

• 대한수의학회지
• /
• 제35권2호
• /
• pp.369-374
• /
• 1995

Aujeszky's disease virus(ADV) was inoculated intranasally into the Korean native goats to investigate pathological findings and pathogenesis of ADV infection by using of histological and immunohistochemical methods and in situ hybridization(ISH). Clinical signs of salvation, pyrexia, pruritus and staggering were followed by death with five days after inoculation, Pathoanatomical findings were edema of the lung and the urinary bladder with hemorrhage and congestion, petechial hemorrhages on the endo-and epicardium, renal congestion, moderate splenomegaly and cystic edema. Main microsocpic lesions observed in all infected goats were confined to the CNS and charcterized by perivascular cuffing with lymphocytes and macrophages, focal gliosis, neuronal degeneration and necrosis, and intranuclear inclusion bodies in the neurons and glial cells. Positive reactions to ADV were detected more frequently in the nuclei than in the cytoplasms of infected nerve cells in the CNS by immunohistochemistry and ISH. Frequenctly localized sites of ADV in the CNS were olfactory bulb, prietal cortex, callosal sulcus and corpus callosum. Positive reactions were also detected in the tonsillar epithelium, and alveolar macrophage and sloughed epithelium of the lung.