1

Speaker(s)：

Wan-Chen Li

Topic

Genetic analysis of H10 subtype avian influenza viruses isolated from wild birds in Taiwan 2013

Abstract

Avian influenza A viruses occasionally cross the barrier to infect species other than birds. The novel H5N1 and H7N9 influenza viruses causing endemics in human are the examples of bird-to-human transmission . To date, highly pathogenic avian influenza (HPAI) viruses belong to H5 or H7 subtypes. HPAI viruses can cause high mortality in poultry, and the viruses spread rapidly and result in huge economic losses. Besides H5 and H7 subtypes, H10 is one of the subtypes to have the potential of threatening poultry or human. There are many records of the H10 subtype viruses infecting poultry or mammalian, and some of the infection cause serious damage or human death. In our surveillance on avian influenza viruses in wild birds, we isolated two strains of H10 subtype virus in 2013. The viruses showed very weak HA genome signal in PCR subtyping system. Genomic sequences of both strains suggested that the strains derived from reassortment of Eurasian and North American lineages.

2

Speaker(s)：

Chieh-Hao Wu

Topic

Molecular Diagnosis of Crayfish Plague and Periodic Report of Electron Microscopy from June to November, 2014

Abstract

The oomycete Aphanomyces astaci is the etiology of crayfish plague and has devastated native crayfish populations across Europe due to introducing the infected North American crayfish. In the end of 2013, the first outbreak of A. astaci infection in cultivated Australian crayfish (Cherax quadricarinatus) was diagnosed in Taiwan. The crayfish specimens were collected from Miaoli, Changhua and Pingtung to establish a diagnosis procedure for crayfish plague. The pathological examinations with special staining revealed penetration of hyphae and granulomatous inflammation in chitinous layer of uropod and articulation. Aphanomyces astaci was not successfully isolated from these specimens based on OIE Manual of Diagnostic Tests for Aquatic Animals. For molecular diagnosis, PCR amplications of internal transcribed spacer (ITS) and chitinase gene fragments were performed and the product sequences analyzed by BLAST showed high identities with A. astaci. In addition, detecting limits between PCR and real-time PCR targeting ITS fragment were compared. By microsatellite genotyping method, genotype D was assumed according to the size of two gene fragments. This study established pathological and molecular diagnoses of crayfish plague.

A total of 217 samples was collected for electron microscopy by negative stain from June to November in 2014, including 39 samples from poultry, 61 samples from herbivores, 32 samples from aquatic animals, 42 samples from swine, and 43 samples for pathogen observation. Forty-five of those specimens were observed with specific viron/bacteria.

3

Speaker(s)：

Chu-Hsiang Pan

Topic

Development of an indirect ELISA for the detection of antibodies to porcine circovirus

Abstract

Porcine circovirus type 2 (PCV2) genome contains two major open reading frames(ORFs), ORF1 encoding the viral replication-associated protein and ORF2 encoding the viral capsid protein. This study was to express the ORF2 capsid protein and Indirect ELISA (iELISA) method was developed for detection of antibody to PCV2. Partial ORF2 gene of PCV2 was amplified by RT-PCR. The RT-PCR amplicons were cloned into a TOPO vector using TA cloning kit; correct clones were confirmed by sequencing. The target genes were subcloned into the pET32 and pCold expression vectors. The expression plasmids were transformed into host cell of BL-21(DE3), then IPTG was used to induce the expression of cloned genes. The solubility of recombinant proteins was tested after induction. Results showed that pCold system expressed a soluble protein and pET32 system showed insoluble inclusion bodies. His tag protein purification kit was used to purify recombinant proteins. The ELISA plates were coated with 2 μg/ml of purified recombinant ORF2 protein and iELISA method was used to test serial dilutions of pig sera which were obtained from the experimental CSFV infected pigs. Results revealed that linear curve could be observed in the iELISA assay. Comparison of homemade and commercial PCV2 IgG iELISA kits for detection of anti-PCV2 antibody from 50 field sera, both of iELISA kits showed high level of correlation (r = 0.938). 

4

Speaker(s)：

Wei-Cheng Hsu

Topic

Report on Attendance of International Meeting " Rabies in the Americas (RITA) " in Mexico

Abstract

The Rabies in the Americas (RITA) meeting is an annual event that has been held since 1990. It has been hosted in many countries across the Americas. For many years, RITA has grown in popularity and prominence with delegates now coming from more than 20 countries across five continents. The meeting provides an opportunity for researchers, health professionals, international, national and local managers of rabies programs, wildlife biologists, laboratory personnel and other people interested in advancing knowledge of rabies surveillance, prevention and control, to meet each other, to share their successes and also to discuss the challenges to be met. Invited by experts in the Centers for Disease Control and Prevention in USA, I attended and presented "Ferret-Badger Rabies in Taiwan" in the 25th RITA meeting held in Cancun, Mexico. By attending the conference, we could update the current global rabies epidemiological situation and the modern rabies studies trend. The information we got had huge contribution to strengthen rabies control and prevention strategy in Taiwan.