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Seminar 781  

Seminar:  781  
1
Speaker(s)
Chun Wan
Topic
Establishment of infectious clone of porcine circovirus type 2
Abstract
Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS). PCV2 infection is also an important infectious disease in swine and sometimes co-infect pigs with other pathogens.  PCV2 was difficultly to be isolated from field samples, and maintained in low virus titer in vitro.  In this study, we have constructed the infectious clone of PCV2 to generate stable virus. The infectious clones were transfected into PK-15 cell line and generate new viral particles.  The highest titer of newly generated viral particles was 105.5TCID50/mL after passage six times.  In addition, there was no significant difference statistically in virus titers generated from various transfect dosages of infectious clone.
2
Speaker(s)
Chia-Yi Chang
Topic
Immunization of swine with developed E2 subunit vaccine of classical swine fever virus
Abstract
Classical swine fever (CSF) is a highly contagious disease of swine. Classical swine fever virus (CSFV) is the causative agent of this disease. CSFV is an enveloped, positive-stranded RNA virus that belongs to the genus Pestivirus within the family Flaviviridae. E2 is the major envelope glycoprotein and represents an important target for the induction of neutralizing immune response against the viral infection. In this study, the efficient and safer marker vaccines weredeveloped by using recombinant E2 protein andsynthesized peptide of E2. Glycoprotein E2expressed in adenovirus transducted PK-15 cells was purified by his-tag column and could be detected by IFA and Western blot. Pigs were immunized with either recombinant E2 protein or peptide. The results of SN test of pig sera showed that neutralizing antibody could be detected from pigs immunized with expressed E2 protein. However, the immune response of pigs with immunization of peptide could not be detected. The challenge of CSFV showed that the immunization of pigs with expressed E2 glycoprotein could induce protection against CSFV, whereas the pigs with immunization of peptide could not survive from the challenge.
3
Speaker(s)
*Tsu-Han Chen, *Yu-Liang Huang, Chu-hsiang Pan, Yeou-Liang Lin
Topic
Taiwan-Thailand Agricultural Cooperation: Evaluation of Diagnostic Kits and Immunological Study on Classical Swine Fever
Abstract
We visited the National Institute of Animal Health (NIAH) and the Regional Reference Laboratory for foot and mouth disease in South East Asia (RRL) forevaluating FMDV non-structural protein (NSP) chromatographic strip and SIV-AIV DNA chip and for study of cell-mediated immunity in CSFV infected pig. Under the Taiwan-Thailand Agricultural Cooperation Project, the technical visit, during July 19th to 24th of 2009, was organized by the Department of Livestock Development, Ministry of Agriculture and Cooperatives, Thailand. To evaluate the chromatographic strip assay for detecting anti-NSP antibodies, 281 pig sera provided by the RRL was tested. Diagnostic sensitivity of the assay in FMDV Asia 1 experimentally infected pigs was 93.8 % (15 / 16), completely agreed with the results obtained from the VIA-AGID and UBI ELISA kits. The results revealed that the assay was able to detect FMDV NSP antibodies elicited by the infection of FMDV serotype Asia1. Sera from pigs vaccinated with trivalent inactivated FMDV vaccine (serotype O, A and Asia 1) and naïve pigs were also used to assess the specificity of the assay. The specificity was 98.1 % (155 / 158) for vaccinated pigs and 100 % (31 / 31) for naïve pigs. Agreement between the results from strip assay and those from the PrioCHECK FMDV-NS kit was 98.6 % (142 / 144). To evaluate the prototype of DNA chip for detecting and genotyping swine and avian influenza viruses, 17 influenza viruses and 8 Newcastle disease viruses provided by the NIAH were tested. The DNA chip demonstrated its ability to identify H1, H3, H5, H6, H7, and H9 subtypes of influenza viruses of either swine or poultry origin. Moreover, H5N1 subtype, a highly pathogenic avian influenza virus, can be simultaneously identified as well. For studying cell-mediated immunity (CMI) in pigs induced by classical swine fever virus (CSFV), we met Dr. Sanipa Suradhat at Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University and worked with her colleagues. Dr. Suradhat presented their achievements and the technology that has been set up in her lab for monitoring CSFV-CMI in pigs. Through this visit, we also learned how to detect CSFV-specific interferon-gamma secreted by peripheral blood mononuclear cells in pigs using ELISPOT or flow cytometry.