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1
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Speaker(s)：
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Lee, Fan
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Topic
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Identification of Culicoidesspecies: entomological training at the National Institute of Animal Health, Japan
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Abstract
In the International Workshop on Arboviral Diseases of Livestock in East Asia, held in Kagoshima, Japan in 2009, the participants agreed to increase exchanges of scientists between the animal health laboratories of the East Asian countries. Following the agreement, a one-week technical visit was made to the Kyushu Research Station of the National Institute of Animal Health in Japan. The aim of this visit is to learn how to collect adult and larval Culicoides in the field and how to identify adult Culicoides by classical entomology and molecular biological technique.
Culicoides biting midges are tiny insects, 1~3 mm in length, and belong to the genus Culicoides within the family Ceratopogonidae. So far, more than 1,400 species of Culicoides have been identified and only a few of them were associated with human and animal diseases. Specific Culicoides species are recognized as biological vectors for arthropod-borne viruses, such as African horse sickness virus, bluetongue virus, bovine ephemeral fever, epizootic hemorrhagic disease virus, Ibaraki virus, Chuzan virus, and Peaton virus. The viruses are transmitted between mammals through biting behaviors of the Culicoides. To control the arthropod-borne virus associated diseases and to monitor the activities of the arthropod-borne viruses in ruminant populations, inclusion of entomological study to the studies of these diseases is a necessity, and establishing capacity to identify Culicoides species of veterinary importance is the first step.
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2
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Speaker(s)：
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Chen, Tsu-Han Coordinator：Lin, Yu-Liang
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Topic
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Development of Luminex assay for detecting the antibodies against foot-and-mouth disease virus non-structural protein
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Abstract
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. Animals infected by FMD virus (FMDV) showed vesicules on the feet, heel bulb, oral cavity, nose and teats. To develop a method with accurate and high sensitivity for the differentiating between the antibodies against FMDV derived from vaccination and those from infection, a Luminex assay was established for the detection of antibodies against non-structural protein of FMDV in our laboratory. A total of 805 pig sera were used for the evaluation of the Luminex assay. The results showed that the sensitivity of the assay was 100 % and the specificity of the assay was 97.5 % ~ 100 %.The agreement between the results from Luminex assay and those from the commercially availableELISAs was up to 90 %. Furthermore, the test results of Luminex assay had a significant correlation with those generated by chromatographic strip and ELISAs. The results revealed that the Luminex assay can be used for the detection of antibodies against NSPs of FMDV.
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3
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Speaker(s)：
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Chen, Yen-Ping
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Topic
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Development of molecular biology diagnostic techniques for Waterfowl Parvovirus
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Abstract
Waterfowl parvovirus can cause diseases with high mortality and morbidity in goslings and ducklings. According to molecular analysis and virus neutralization tests, waterfowl parvovirus can be divided into the goose parvovirus (GPV)-related group and the Muscovy duck parvovirus (MDPV)-related group. In Taiwan, both GPV and MDPV have existed in the field and the disease caused by waterfowl parvovirus is controlled using live attenuated vaccines. In this study, we developed PCR method to distinguish GPV from MDPV in one tube reaction with sensitivity of 100 copies when existing only nucleotides of GPV or MDPV and with sensitivity of 1,000 copies when existing both of nucleotides of GPV and MDPV. We also developed PCR method to differentiate the wild and vaccine strain of GPV. The size of PCR products amplified from wild strains was 20 nucleotides larger than those amplified from vaccine strains. We also analyzed the complete nucleotide sequences of viral protein 1 (VP1) genes of GPV isolated from 2006 to 2010 in Taiwan. The VP1 nucleotides of recent 5-year isolated GPV showed 99.3% - 100% identity with each other and showed only 0.4% - 0.6% difference with GPV strain isolated in 1982 (82-0321). The results indicate that the VP1 genes of GPV had remained highly stable in the field in Taiwan.
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4
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Speaker(s)：
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Tsai, Kuo- Jung
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Topic
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The report of “learning the techniques for diagnosis and surveillance of bat lyssavirus by visiting CDC in USA” and “the surveillance of bat rabies in Taiwan”
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Abstract
A researcher is dispatched to visit CDC, Atlanta and learn the bat lyssavirus diagnosis and surveillance technique from October 31 to December 7 in 2010 to implement the international cooperation project, “establishing the surveillance and prevention system of potential and emerging zoonotic diseases”. During the period in Atlanta, the researcher learn the rapid fluorescent focus inhibition test (RFFIT), reverse transcription-polymerase chain reaction (RT-PCR) and examine the bat serum samples collected in Taiwan in 2010. The above techniques can be applied to the diagnosis and surveillance of rabies and lyssavirus to serve as disease screening and early warming. To conduct the bat rabies surveillance, bat brains (n= 187) were collected from 2008 to 2009 and subjected to the direct fluorescent antibody (DFA) test. No brains were showed rabies or lyssaviral antigen. Bat sera (n= 60) were sent to CDC, Atlanta where RFFIT was conducted to detect the antibody against 5 different lyssavirus including Australian bat lyssavirus. And all serum samples were negative. In 2010, brains of 41 bats are collected and subjected to the DFA test and all are negative. Meanwhile, bat sera (n=75) are collected and sent to CDC, Atlanta where the dispatched researcher examine antibody against 2 different lyssavirus, Irkut virus and Khujand virus using RFFIT technique under the guidance of expert in CDC, Atlanta. All serum samples are negative. The antibody examination for other lyssavirus will be finished by expert in CDC, Atlanta. The report describing the surveillance of lyssaviral antigen and antibody in bats in Taiwan can serve as an epidemiologic surveillance data for rabies- free status of Taiwan. We look forward to advanced cooperation between USA and our country on the topics for emerging zoonotic disease of wildlife.
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