PA-01

Aspirin and AICAR Suppress Bovine ephemeral fever virus-Induced Autophagy via Downregulation of the PI3K/Akt/NF-κB and Src/JNK Pathways Inhibiting Virus Replication

Hsu-Hung Tseng1, 6, Ching Y. Cheng1, Hung-Chuan Chiu1, 3,Ming H. Wu1, Ching-Dong Chang2 and Hung-Jen Liu1,3, 4, 5

1Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan

2Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan

3The iEGG and Animal Biotechnology Center, National Chung Hsing University, Taichung, Taiwan

4Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung, Taiwan

5Ph.D Program in translational Medicine, National Chung Hsing University, Taichung, Taiwan

6Division of General Surgery, Taichung Hospital, Ministry of Health and Welfare, Taichung, Taiwan

Autophagy is the cellular catabolic processes in which cytoplasmic target material is transported to lysosomes for degradation. Many viruses have encoded autophagy-promoting proteins to assist their survival in their host cells. In this work, we report for the first time that BEFV infection and BEFV M transfection are able to induce cellular autophagy via upregulation of the Src/JNK, PI3K/Akt/NF-κB pathways and downregulation of the PI3K/Akt/mTOR pathway at the middle to late stage of infection. In the present study, we have demonstrated that aspirin and 5-aminoimidazole-4-carboxamide-1-b-riboside (AICAR) inhibit BEFV replication. The mechanisms underlying aspirin and AICAR impacting host cells to inhibit BEFV replication were elucidated. AICAR suppresses the PI3K/Akt/NF-κB and Src/JNK pathways via inhibition of PGE2/E-prostanoid (EP) receptor 2, thereby reducing BEFV-induced autophagy and levels of LC3-II while aspirin blocks only the PI3K/Akt/NF-κB pathway, thereby reducing LC3-II level. Interestingly, AICAR inhibits BEFV-induced autophagy in an AMPK-independent manner. In addition to downregulation of the Src/JNK pathway, we also uncovered that AICAR inhibits BEFV-induced expression of ULK1, Atg7, and LC3. Moreover, degradation of Atg7 caused by AICAR could be restored by MG132, suggesting that BEFV-mediated Atg7 degradation via the ubiquitin-proteasome pathway. Taken together, both AICAR and aspirin have potential as antiviral drugs against BEFV infection.

Keywords: Autophagy, AICAR, aspirin

 

PA-02

Evaluation of the Biosecurity Risks of Porcine Reproductive and Respiratory Syndrome Infection by COMBAT System

Chun-Wei Liu1, Chao-Nan Lin1 and Ming-Tang Chiou1,2

1Department of Veterinary Medicine, National Pingtung University Science and Technology, Taiwan

2Research Center for Animal Biologics, National Pingtung University Science and Technology, Taiwan

Porcine reproductive and respiratory syndrome virus (PRRSV) causes economic loss in swine industry around the world. Preventing PRRSV infection is mainly based on vaccines and biosecurity management. There are many commercial vaccines available in Taiwan. However, efficient controlling of PRRSV is still a big challenge. To find out the relationship between biosecurity and PRRS infection, five PRRSV contaminated farms implemented PRRSV vaccination were investigated. The COMBAT system developed by Boehringer Ingelheim was applied to evaluate the biosecurity risks in these farms. In the COMBAT system, fifty-five questions, including four parts internal, external, management and location, were answered by each farm and further analyzed to calculate the degrees of different kinds of biosecurity risks. Due to the characteristics of farm scales and densities in Taiwan, the external and location risks of all farms were high. The internal and management risks varied based on the immune programs and management approaches in different farms. The biosecurity of these farms is defective and may result in a major loophole of PRRS prevention.

Keywords: COMBAT, porcine reproductive and respiratory syndrome virus, biosecurity

 

PA-03

Inhibiting Capacity of Chlorine Dioxide (ClO2) on Ornamental Fish Diseases

Chia-Te Lin1 and Shih-Chu Chen1,2

1International Degree Program of Ornamental Fish Science and Technology, International College, National Pingtung University of Science and Technology, Pingtung, Taiwan

2Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan

Due to Taiwan aquaculture industry's intensive professional aquaculture mode, farmers, sales and aqua enthusiasts rely on lots of drugs to control and prevent the spread of fish diseases. However, fish diseases are resistant to drugs gradually, and drug residues can cause environmental disadvantage. Many researchers try to seek effective solutions for sustainable development. The purpose of this study is to apply chlorine dioxide, a disinfectant, to the aquaculture industry to test the impact on fish diseases and fish health. From January 2018 to January 2019, two experiments were conducted. Experiments were conducted in a government-approved tertiary laboratory and adhered to relevant experimental specifications. In the first experiment, different concentrations were determined for the three pathogenic Vibrio species (Vibrio alginolyticusVibrio parahaemolyticusVibrio harveyi) isolated from 12 cases of diseased amphiprioninae fish (clownfish). The results were estimated using the CFU plate culture method. In the second experiment, according to the results of the first experiment, the appropriate CLO2 concentration was designed to carried out Lethal Concentration 50% (LC50) test on the anemone fish. Observe epidermis and internal organs of the fish were damaged or mutated to find the best concentration, as a daily protection and disease treatment application. The experimental results show that when the concentration is increased to 3.75 ppm for 10 minutes, the three kinds of Vibrio cannot survive. For clown fish, Lethal Concentration 50% when the concentration reached 0.7125 ppm.

Keywords: amphiprioninae fish, Vibrio, chlorine dioxide, ornamental fish

 

PA-04

Ultrasound Biomicroscopic Study of Anterior Segment Changes after Phacoemulsification and Intraocular Lens Implantation in Dogs

Kai-Ting Hsu and Shiun-Long Lin

Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan

Ultrasound biomicroscopy allows for qualitative evaluation in the structures of the anterior segment as well as for quantitative measurements of anterior chamber components. Previous research in humans indicated that the capsular bag stretched horizontally with reduced vertical diameter as a result of adaptation to the implanted intraocular lens after cataract surgery. The purpose of this study was to measure the intraocular status in the dogs after phacoemulsification with lens implants and to compare preoperative and postoperative changes. Twenty eyes of 19 dogs with different cataract stage and no history of previous ocular hypertension or glaucoma presented to the ophthalmology Department at the NCHU-VMTH for phacoemulsification surgery between November 2017 and October 2018. All patients were examined with UBM and A-mode before and 1 and 2 and 4 weeks after phacoemulsification surgery. At each UBM examination, axial images of the anterior chamber and radial sections of the angle at the superior and lateral quadrants were obtained. The parameters were anterior chamber depth (ACD), axial length, capsular bag diameter, angle opening distance, anterior chamber angle and angle recess area. The mean ACD after surgery was 6 mm deeper than previous research and showed statistically significant change after surgery but showed no associated with intraocular pressure and axial length. The values correlate fairly well with previous study and further support the idea of anterior chamber changing. In conclusion, phacoemulsification with lens implants will change anterior chamber and iridocorneal angle.

Keywords: ultrasound biomicroscopy, phacoemulsification, iridocorneal angle, intraocular pressure, angle opening distance

PA-05

Establishing E. maxima Wild Strains Lesion Scores Standard Challenge Amount

Yu-Sao Chang

Department of Veterinary Medicine, National Pingtung university and Technology, Pingtung, Taiwan

Chicken coccidia is a parasite disease can cause chicken death, feed conversion rate(FCR) performance poor and extra economic losses. E. maxima, E. tenella, E. acervulina are the most common coccidia in chicken farm. The purpose of this study is to establish E. maxima 's 4 lesion soring levels standard challenge amount for the further experiment. The classification of the lesion level was mainly judged by the lesions of the intestinal wall and the intestinal contents condition. Intestinal wall thickening and emphysema, intestinal serosal surface hemorrhage when the lesion get severe will enlarge to ecchymosis. The intestinal contents depends how serious of infection, low level infection is watery and clear mucus, high level infection is orange thick mucus and putrid odour. Depends on classification to these lesions. We established the oocysts challenge amount reach lesion scoring level 1 require 5.0 × 104/bird, lesion soring level 2 require 1.0×105/bird, lesion soring level 3 require 2.5×105/bird, lesion soring level 4 require 4.0×105/bird

Keyword: Chicken coccidia, E. maxima, Lesion Scores

 

PA-06

Avian Reovirus p17 Protein Disrupts mTOR Complex 1 Assembly by Enhancing the Binding of PRAS40 and FKBP38 to mTOR Complex 1 and by Negatively Regulating the Akt/mTORC1 Pathway Inducing Autophagy and Increasing Virus Replication

Jyun-Yi Li1, Wei-Ru Huang1,2, Ching-Dong Chang3 and Hung-Jen Liu1, 2, 4, 5

1Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan

2The iEGG and Animal Biotechnology Center, National Chung Hsing University, Taichung, Taiwan

3Department of Veterinary Medicine, National Pingtung University of Science and Technology

4Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung, Taiwan

5Ph.D Program in translational Medicine, National Chung Hsing University, Taichung, Taiwan

Avian reovirus (ARV) non-structural protein p17 affects cellular function and regulates signal transduction. The mammalian target of rapamycin (mTOR) signaling pathway is a regulatory center that coordinates cell growth, proliferation, survival, and metabolism in eukaryotic cells. mTORC1 is a well-studied negative regulator of autophagy. The ARV p17 protein-mediated Akt inhibition and activation of the p53/PTEN pathway results in suppression of Akt phosphorylation at S473 and T308. The present study demonstrates that p17 disrupts mTORC1 assembly by enhancing both PRAS40 binding to raptor and FKBP38 binding to mTORC1 to suppress mTORC1. Inhibition of PRAS40 phosphorylation by p17-mediated Akt inhibition suppresses mTORC1 translocation to lysosome, impairing mTORC1 activated by Rheb-GTP in lysosome. Overexpression of PRAS40 (T246A) mutant further enhanced p17-mediated suppression of mTORC1 while overexpression of Rheb reversed the effect of p17 on suppression of mTORC1. The present study provides mechanistic insights into p17-mediated suppression of mTORC1, which induces cellular translation shutoff and autophagy, benefiting virus replication.

Keywords: Avian Reovirus p17, mTOR Complex 1, PRAS40