107 年度春季研討會 壁報論文競賽組 (PB)

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PA-01

Identification and Characterization of the Serotype-determining Sequences of the Hemagglutinin Protein from Avibacterium paragallinarum

Shu-Wan Huang1, Jui-Hung Shien2, and Poa-Chun Chung1

1Graduate Institute of Microbiology and Public Health, National Chung Hsing University,

2Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan

Avibacterium paragallinarum is the causative agent of infectious coryza, an important acute respiratory disease associated with growth retardation and reduced egg production in egg-laying hens. The hemagglutinin (HA) protein plays a key role in virulence and immunogenicity of Av. paragallinarum. HA has been subtyped into three serovars: A, B, and C. Though hemagglutination-inhibition (HI) assay is the golden standard for serotyping of Av. paragallinarum, this method is complicated and technically demanding. The purpose of this study is to identify the serotype-determining sequences of the HA protein and use phylogenetic analyses to identify the serotypes of Av. paragallinarum. Towards this goal, we constructed strains of Av. paragallinarum that express a series of hybrid HA proteins containing different portion of HA sequences from strain 221(serotype A) and strain TW07 (serotype C). HI tests using HA antigens prepared from these strains revealed that the amino acid sequences that determine the serotypes of A and C are located at residues 240-614 of the HA protein. We then used prokaryotic system to express a recombinant protein containing HA240-614 sequences. Western blot analyses showed that anti-serum against strain 221 can distinguish between HA240-614 from strain 221 and that from strain TW07. Serotyping of different Taiwanese field strains of Av. paragallinarum using phylogenetic analysis of HA240-614 sequences is consistent with that of HI testes. This study is the first report showing that HA240-614 contains the serotype-determining sequences of Av. paragallinarum. This finding can be used to develop a method for rapid serotyping of strains of Av. paragallinarum.

Keywords: Avibacterium paragallinarum, serotype-determining sequences, hemagglutinin

 

PA-02

Safety Assessment of Edible Oil Before and After High Temperature Processing by Medium-Term Hepatocarcinogenesis in Rats

Yun-Chieh Tuan1, Yi-Hui Su1, Yan-Xiu Lin1, Tony Ji Fang2, Jiunn-Wang Liao1, 3

1Graduate Institute of Veterinary Pathobiology, National Chung Hsing University, Taichung, Taiwan R.O.C

2 Food Industry Research and Development Institute, Hsinchu, Taiwan R.O.C.

3Animal Disease Diagnostic Center, National Chung Hsing University, Taichung, Taiwan R.O.C.

Nowadays, news that focus on health effects, including carcinogenesis, caused by eating the fried foods has been reported more frequently, which raised the question that whether long-term intakes the fried foods increase the risk of cancer? Therefore, the aim of this study is to evaluate the potential carcinogenicity of oils via medium-term bioassay model in rats. 8 week-old, SD rats, 6 males per group were used. This experiment was divided into five groups: Blank Control (BC), Initiator DEN Control (DC), Positive Control (PC), UHOil and HOil groups. Firstly, Diethylnitrosamine (DEN) 200 mg/kg was intraperitoneally injected on day 1 as an initiator. At the second week, administered the samples by gavage in UHOil and HOil groups, and added 2-AAF 200 ppm in diet in the PC only, and oil samples were gavaged for the consecutive 7 weeks. At the third week, 1/3 partial hepatectomy was performed to be a promoter. The body weight and food consumption, urinary, complete blood counts, biochemistry, gross and histopathology were recorded at the end of experiment. In addition, the tumor marker of GST-P was detected in liver by immunohistochemistry (IHC) and Western blot analysis for the carcinogenicity. Except rats in the PC was successfully induced hepatoma, neither the UHOil nor HOil-treated groups posed hepatocarcinoginicity in male rats. Based on the above results, both the unheated and heated edible oils are classified as “not carcinogenic” in the medium-term bioassay model in rats.

Keyword: GST-P, heated, carcinogenicity, medium-term bioassay, soybean oil, rats

 

PA-03

Retrospective Pathological Reports of Splenic Tumors in Domestic Hamsters

Yun-Chieh Tuan1, Ruo-Chan Wang2, Ju-Po Kao2, 3, Kimimasa Takahashi4, Jiunn-Wang Liao1, 5

1Graduate Institute of Veterinary Pathobiology, National Chung Hsing University, Taichung, Taiwan R.O.C

2Veterinary Medical Teaching Hospital, National Chung Hsing University, Taichung, Taiwan R.O.C.

3Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan R.O.C.

4 Department of Veterinary Pathology, Nippon Veterinary and Animal Science University, Musashinoshi, Tokyo, Japan.

5Animal Disease Diagnostic Center, National Chung Hsing University, Taichung, Taiwan R.O.C.

A total of 15 splenic biopsy specimens in 212 cases of hamsters from the Division of Wild (Exotic) Animal Medicine, Veterinary Medical Teaching Hospital, National Chung Hsing University, between 2010 and 2017 were studied for the retrospective pathological study. The incidence in the spleen was 7.1% (15/212), including 15 Phodopus hamsters. Ten neoplasms and 5 non-neoplastic lesions in the Phodopus hamsters occurred. For neoplastic lesions, the prevalence in Phodopus hamsters was higher than in Syrian hamsters (found 3 cases in VMTH, NCHU), which are similar to laboratory hamsters. In Phodopus hamsters, the most common tumors are histiocytic sarcoma (HS), lymphoma, malignant fibrous histiocytoma (MFH) and hemangiosarcoma. In addition, hemangiosarcoma and histiocytic sarcoma were found in Syrian hamsters. The non-neoplastic lesion was a fibrotic nodule. The mean age of affected hamsters was 16.6 months, and females were affected more than males. In immunohistochemical staining, histiocytic sarcoma and malignant fibrous histiocytoma were positive for lysozyme, lymphoma was negative for CD79a and CD20, one case of lymphoma was positive for CD3, and hemangiosarcoma was positive for Von Willebrand factor (VWF). In terms of non-neoplastic lesions, the most common are fibrotic nodules and all occur in aging female hamsters. The nodules consist of collagen fiber that can be identified with Masson's trichrome stain, and are related to repairing of trauma in the spleen. To our knowledge, there have been no retrospective pathological reports of splenic tumors in domestic hamsters. Therefore this study can provide some valuable analytical data about spleens in hamsters.

Keyword: tumors, non-neoplastic lesion, retrospective pathological reports, spleen, hamsters


 

PA-04

Development of Porcine Epidemic Diarrhea Virus Particle-Type Oral Vaccines by Using Sodium Alginate and Chitosan as Biological Materials

Lian-Xiu Liu1, Yao chi Chung1

1Graduate Institute of Animal Vaccine Technology, National Pingtung University of Science & Technology

The porcine epidemic diarrhea virus (PEDV) particle-type oral vaccines can be more convenient and safe than traditional vaccine, and can reduce the stress caused by administration by injection, also can effectively induce the better mucosal immune response than that induced by conventional injection. PED mainly caused by fecal infection. The infected pigs will cause diarrhea, vomiting, and even death by dehydration. Therefore, this study used chitosan and sodium alginate to develop particles-type oral vaccine that could induce intestinal mucosal immunization of pigs to against PEDV infection. The chitosan and sodium alginate are both biodegradable and not toxic to the immunized animals. In this study, we cloned the PEDV S protein which is able to induce neutralizing antibody response as target antigen. Preliminary results have confirmed the size and antigenicity of protein products using SDS-PAGE and Western blotting. The particle size and shape integrity of particle-type oral vaccines were analyzed by using scanning electron microscopy. After the protein was coated in oral vaccines, the amount of coated protein, coating efficiency, and release efficiency were analyzed. Animal test were performed by oral immunization with mice, the protective efficacy of the vaccine was evaluated by ELISA antibodies, cytokine expression.

Key words: particle type vaccine, Chitosan, Sodium alginate, porcine epidemic diarrhea virus, S protein, mucosal immunity

 

PA-05

Study on Antibiotic Susceptibility Test to Riemerella anatipestifer and E. coli in Waterfowl

Ding-Bang Huang1, Chia-Ying Lin1, Ching-Dong Chang1, Shyh-Shyan Liu1, Chin-En Tsai1, Hung-Yi Wu1

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

In 2015,the output value ofthe waterfowl industry was approximately NT$10 billion dollars, and it was mainly in Pingtung County, Changhua County, and Yunlin County.Among them,breeding in Pingtung County was the largest, accounting for one-third of the total.Due to the serious abuse of antibiotics by farmers, the bacterial resistance caused serious economic losses, This experiment counts the results of antibiotic susceptibility test of Riemerella anatipestifer and E.coli from 2016-2017 years to provide veterinarian clinical medication reference.The specimens were collected on the liver and air sac of ducks and geese to bacterial culture and antibiotic susceptibility test. Rimererella anatipestifer (33%, 92/282) was the most common and E.coli (23%, 62/282) Mixed infection (6%16/282) was using by polymerase chain reaction. Riemerella anatipestifer is most sensitive to Cephalexin (48%), Tilmicosin (84%) is the highest resistance, Trimethoprim Sulfamethxazole (83%) is the second. E.coli is most sensitive to colistin (36%)Cephalexin(35%), Tilmicosin(97%) is the highest resistance. Riemerella anatipestifer mainly occurs in winter, occurs in 3-10 weeks waterfowl. E. coli can occur at all ages and all years in waterfowl. Occurrence of Riemerella anatipestifer in the case of polyserositis was (68%). The incidence of E. coli was (47%). Riemerella anatipestifer is most effective with Cephalexin, and E. coli is most effective with colistin.

Keywords: Waterfowl, Riemerella anatipestifer, E.coli, Antibiotic Susceptibility Test

 

PA-06

A Study on Probiotic from Crops of Parrots and Pigeons

Yu-Cheng Chang1, Chia-Ying Lin1,Ching D. Chang1, Shyh-Shyan Lin1, Chin-En Tsai 1, Hung-Yi Wu

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

According to the supervisor of Kaohsiung County Bird Watching Promotion Association in 2011, in Taiwan, parrots accounted for 70% of the ornamental bird market and the export value reached as high as 7 billion. Among a variety of bird diseases, the lesions of the crop are one of them; the crop is a chamber in the middle part of the esophagus of some birds, and functions include the storage of food bolus and the decomposition of nutrients by the probiotics. In clinical practice, cases associated with crop contain burns and inflammation of it due to the high temperature of the fed powdered milk. In many cases, the bacteria in the crops have become chaos, causing food groups to stagnate and failing to enter the stomach, causing indigestion and secondary infections. In the paper BERGEY'S MANUAL OF SYSTEMATIC BACTERIOLIGY, M. AndreÍna Pacheco indicated that the strains in the parrots that prefer to eat cereals include Enterococcus, Lactobacillus, Streptococcus, Propionibacterium and Enterobacteria Enterobacteriaceae, in which Lactobacillus have the ability to secreted amylases in order to decompose foods, this reserch want to develop Lactobacillus, which belongs to parrots and pigeons, by understanding the main species of normal lactobacilli in the crops of parrots and pigeons. In this study, the API 50 colonization system was used. We found that Lactobacillus, a common bacillus in the parrot crop, contained Lactobacillus (A), Lactobacillus (B), Leuconostoc (C), Lactobacillus (D), and Lactobacillus (E). Lactobacillus (F) and Lactobacillus (G) are found in the crops of pigeons. Two (33%) samples presented Lactobacillus (A) positive individuals, Positive individuals of Lactobacillus (B), Leuconostoc (C), Lactobacillus (D), and Lactobacillus (E) were each (17%); among them, Lactobacillus (B) and Lactobacillus(C) were detected in the crop one individual. In pigeons, we detected Lactobacillus (F) (50%) and Lactobacillus (G) (50%) in 2 individual samples. This study found that the unique Lactobacillus species of several parrots and pigeons are developing toward clinical applications in the future.

Keywords: Parrot, Pigeons, Crop, Lactobacillus

 

PA-07

Health Monitoring and Analysis of Formosan Sika Deer (Cervus nippon taiouanus) in Sheding Village of Kenting National Park, 2017

Yu-Hsiang Chen1, Chia-Ying Lin1, Ching-Dong Chang1, Shyh-Shyan Liu1, Chin-En Tsai1, Hung-Yi Wu1

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

The Formosan Sika Deer (Cervus nippon taiouanus), a subspecies of Sika Deer (Cervus nippon), is an endemic species of Taiwan. Since 1984, the Formosan Sika Deer restoration program in Kenting National Park has resulted in the current population size, exceeding 1500 individuals, also, the Sheding Rehabilitation Center has perpetuated the purebred traits of Formosan Sika Deer. In the year 2017, Sheding Rehabilitation Center recalled two groups (a total of 70) of Formosan Sika Deer for the evaluation of individual health conditions. A jugular venous sampling section was carried out, after an immobilization was performed with plastic projectile syringes blown from a blow-pipe, and an analysis on hematological and serum biochemical values was done. For the monitoring of parasites, a parasitic oocyte inspection on fecal samples (randomly sampled) was performed. An Intradermal Tuberculin Test (ITT) was performed on the neck area to monitor Tuberculosis. The results were negative for both parasitic oocyte inspection and ITT. The analysis of hematological and serum biochemical values showed that a number of individuals has shown a significant difference in their values; 50% of individuals was diagnosed with physiologic leukocytosis (mild neutrophilia with normal lymphocyte count or mild lymphocytosis) which might be caused by a stressful capture; 5.71% of individuals was analyzed with a higher packed cell volume (PCV), hemoglobin (Hb) and red blood cell count (RBC) which might be caused by stressors (e.g. immobilization by dart-blowing and blood sampling), taken measures (e.g. food and water limitation) which lead to dehydration; there were 17.14% samples analyzed with an increased mean corpuscular volume (MCV); blood glucose level was low in 2.85% of the samples, and a slight decrease in blood total protein in 4.29% of the samples; there were no simultaneous increase in both BUN and CRE, an increase in either BUN or CRE might be caused by temporary dehydration due to captivity; the microfilaria infection of Setaria spp. was detected in 1.43% of the samples. From the findings, the overall status of the Formosan Sika Deer was fair, although a minority was influenced by stress factors, low nutritive condition and microfilaria infection.

Keywords: Kenting National Park, Formosan Sika Deer, Health care, Hematology, Serum Biochemistry

 

PA-08

Sterilization Effect of Nano-silver in Geese Against Bacteria

Wei-Lin Shih1, Chia-Ying Lin1, Ching-Dong Chang1, Shyh-Shyan Liu1, Chin-En Tsai1, Hung-Yi Wu1

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

Goose is one of the important agricultural industries in Taiwan, the total number of geese slaughter in 106 years has grown 52.95% compared with 105 years, which shows that the industry is recovering. However, the common problem of bacterial infection has always existed, the genus Salmonella spp. also known as Goose paratyphoid, is a common infectious disease of all kinds of poultry, mainly endanger the young geese, and often through contaminated animal products infect human beings, endanger people and animals very heavy. In the face of bacteria most of the use of antibiotics, not only to worry about the withdrawal of antibiotics, but also may cause abuse of antibiotics, resulting in bacteria to produce drug resistance. Therefore, Nano-silver, which has less influence on environment and biology, is chosen as the subject of experiment. Nano-Silver special sterilization mechanism makes the bacterium not easy to produce the resistance, the silver ion can with the bacterium protein the mercaptan base (-SH) the specific binding force to cause the bacterial protein denaturation, achieved the sterilization function. So, we want to discuss the antibacterial effect of nano-silver on salmonella and geese in various concentrations and at different time points. The study found that the minimum effective dose of nano-silver was 20ppm, and then the nano-silver was used in the same concentration of 10ppm and 100ppm in geese to observe the total amount of feces. Simulation of the effect of nano-silver prevention, the owner of the goose to the silver nanoparticles, due to climatic factors, found that the whole field of bacterial infection, after testing determined, began to trace the total amount of feces, to record whether the nano-silver can effectively inhibit bacteria, the experiment proved that nano-silver in the use of the fourth day could be reduced by 43%. Simulation of the effect of nano-silver treatment, the owner of the test to determine the goose farm has a bacterial infection, and then cast nano-silver, we go to the scene to sample, test the total amount of feces changes. Found that in the treatment of no more than the effect of prevention, the highest can only inhibit the total number of bacteria 8%, the understanding of whether the nano-silver residue in the geese, nano-silver to give one months after the owner of the test, the results of our experimental nano-silver will indeed remain in the body.

Keywords: nano-silver, goose, minimum effective dose, prevention, treatment


 

PA-09

The Investigation of Cryptosporidium Infection in Dairy Cows and Calves in Taiwan

Jui-Yu Lee1, Hsu-Hsun Lee1

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

There are four species of Cryptosporidium commonly reported in cattle; C. parvum, C. bovis, C. ryanae and C. andersoni. C. parvum has the highest pathogenicity among them. The most common clinical sign is diarrhea and causing considerable direct and indirect economic losses. Former studies indicated that calves are susceptible hosts to C. parvum. Moreover, recent studies pointed out the high possibility of calves infected Cryptosporidium right after being delivered due to the carrying of this pathogen by their dams. This research will divide Taiwan into four area and conduct the investigation of the prevalence of Cryptosporidium infection and the number of the oocysts in the environment in the expectant dairy cows and calves using nested polymerase chain reaction (nested PCR). The sampling will focus on feces, drinking water and flooring sample. After DNA extraction, nested PCR will be used to detect Cryptosporidium and the product will send to DNA sequencing. The infection rates of southern Taiwan were 32% (63/197), which is significantly higher (P<0.05) than the infection rates of middle Taiwan (21.5%, 26/121) and extremely significantly higher (P<0.01) than the rate of eastern Taiwan (11.7%, 10/86). No significant differences of environment positive rates were found among four areas of Taiwan. The farm positive rates of calves’ flooring sample was 70% (14/20), which is significantly higher (P<0.05) than the farm positive rats of expectant cows’ flooring sample (15%, 3/20). So, we concluded that calves’ environment plays an important role in calves’ cryptosporidiosis.

Keywords: Cryptosporidum, dairy cow, environment, prevalence

 

PA-10

Epidemiology of Bartonella, Anaplasma and Borrelia burgdorferi Infections among Small Mammals, Carnivores and Non-human Primates in Taiwan.

Yu-Ting Lo1, Chen-Chih Chen2, Chuen-Fu Lin3, Chao-Chin Chang4,*

1,4Graduate Institute of Microbiology and Public Health, National Chung Hsing University, Taichung, Taiwan

2Wildlife Conservation Institute, National Pingtung University of Science and Technology, Pingtung, Taiwan

3