The occurrence and chemical control of downy mildew on Eustoma graizdtflorum Salisb

1993年1月中旬於南投縣埔里鎮及仁愛鄉清境農場之簡易溫室內發現洋桔梗露菌病之發生，旋於屏東縣鹽埔鄉亦陸續發現本病之蹤跡，至3月中旬，幼苗及成株皆發病嚴重，病徵最初出現於嫩葉，葉片下表皮產生灰色黴狀物，並逐漸擴展，罹病部份葉片因而稍呈褪色現象，葉片並向下扭曲，隨著病勢進展，葉片扭曲現象增加，嚴重時葉片上產生淡褐色病斑，其上覆蓋病原菌之菌體，若不加防治則整株枯死。原菌Peronospora chlorae de Bary，胞囊柄雙叉分枝，由寄主組織表面抽出，胞囊呈橢圓形，大小為16-22×10-15µm，平均為19×12.5µm，著生於胞囊柄頂端；胞囊直接抽出發芽管發芽，於8-32℃溫度範圍內，發芽率無差異。自日本進口之種子在臺灣播種可明顯發現感染露菌病，七個品種之罹病率介於12.46～35.63％之間。田間藥效試驗進行期問各藥劑均未發現藥害產生，然藥效間顯示5％顯著差異，推薦35％腈硫克絕可濕性粉劑1200倍、80％福賽得可濕性粉劑800倍、64％甲鋅歐殺斯可濕性粉劑400倍、66.5％普拔克溶液800倍及35％本達樂可濕性粉劑2000倍為防治洋桔梗露菌病之藥劑；然35％腈硫克絕可濕性粉劑1200倍於苗期施用時，明顯抑制種子發芽及幼苗生長，故不宜施用於苗床期。 The downy mildew in the vinyl house both in Puli and Jenai area was found occurring on Texas bluebell (Eustoma grandiflorum Salisb.) in mid January, 1993. The disease was also found at Yenpu near Pintung city at the same time. The disease causes severe damage on both seedlings and adult plants. The small gray mycelial masses appeared on the lower surface of young leaves and disseminated to make a inconspicuous discolored spots. The leaves curled to the lower surface of leaves and became twisted and distorted. And the spots then changed to slight brown and covered with mycelial mass and spores. Finally the infected leaves turned to dark brown and the whole plants died in severe condition. The causal organism was Peronospora chlorae de Bary. The erect sporangiophores on lower leaf surface are dichotomously branched with sporangia borne on sharply pointed terminal branches. Sporangia are 16- 22 X 10-15µm (19 X 12.5 µm in average) and ellipital in shape. They germinated directly to form germ-tube at 8-32℃. The disease incidence was 12.46- 35.63% when seven varieties imported from Japan were surveyed during seedling bed. The primary inoculum of downy mildew was suspected to be coming from seeds. The fungicidal efficacy was various in protecting plants against the pathogen and no phytotoxicity was found when screening test of fungicides was conducted in the fields. Five kinds of fungicides, 35% cymoxanil +dithianon WP in 1:1200 dilution, 80% fosetyl-Al WP in 1:800 dilution, 64% propineb +oxadixyl WP in 1:400 dilution, 66.5% propamocarb hydrochloride S in 1:800 dilution and 35% bena]axyl WP in 1:2000 dilution showed the best efficacy in field trials. The cymoxanil + dithianon 35% WP was not recommended to use in the seedling beds for the reason that the germination of seeds was inhibited apparently after treatment

The Survey of Postharvest Diseases and the Controlling Methods of Citrus and Some Other Fruits

依據國外報告，柑桔貯藏病害以綠黴病、青黴病、炭疽病、黑色蒂腐病、褐色蒂腐病、果腐病、黑腐病、褐腐病、灰黴病及菌核病等為主；臺灣累積三年之調查結果，亦以綠黴病、青黴病、炭疽病、果腐病、黑腐病為主，而由Botryosphaeria sp.引起之蒂腐病亦相當普遍，至於灰黴病及菌核病則甚少發現。不同種之柑桔對貯藏病害之感病性有差異，其中以甜橙最為感病。田間栽培管理、採收及貯運方式明顯影響貯藏病害之發生，至於採收時若遇陰雨，則往往導致貯藏病害之大發生，乃因採收過程中造成之傷口為病原菌之主要侵入管道，而下雨造成之高濕度則為病勢進展之助力，因此改變採收及貯運方式以減少傷口，可降低貯藏病害之發生。而採收期間之藥劑處理以採收後浸果處理之效果最佳，但農藥殘留量亦高。貯藏空間之環境衛生、罹病果實及包裝材料之清理亦明顯影響貯藏病害之發生；因此於栽培期加強管理減少病害發生、採收時避免傷口產生、採收後適度藥劑處理，再配合貯藏前貯藏空間之廢棄物清理及消毒、貯藏期間定期清理病果及殺菌，當可減少貯藏病害之發生。 Ten kinds of postharvest diseases were occurred severe on the citrus cultivars when survey was done in the foreign country. They were green mold, blue mold, anthracnose, diplodia stem- end rot, phomopsis stem-end rot, fusarium rot, alternaria rot, brown rot, gray mold and cottony rot. Five kinds of postharvest diseases were occurred frequently in Taiwan, when surveyed continuously within recently three years. They are green mold, blue mold, anthracnose, fusarium rot and alternaria rot. The disease caused by Botryosphaeria sp. also occurred commonly. The gray mold and sclerotinia are found scarcely in Taiwan. The variety sweet orange( Citrus sinensis Osb.) was the most severe when different varieties were store and showed different degree of sensitivity to postharvest diseases. Both the cultural practice and methods of harvesting and transporting could influence the occurrence of postharvest diseases. The injury produced during the harvesting and handling period could induce the severity of postharvest diseases if the fruit were harvested in the raining days. The major reason is because of the high humidity for disease to development and the infection occurs commonly through injuries inflicted during harvesting and handling. The dipping method showed more effective than other methods when fungicides were used and high residue problem was remained. Another three factors could influence the occurrence of postharvest disease significantly. They were the sanitary of packinghouse, number of diseased fruits and cleaning of the packing material. The strategy for controlling postharvest disease followed step by step are good cultural practice, avoiding injury formation during harvesting, treatment of chemicals after postharvesting, and sanitary of packinghouse and packing material

Integration Crop Management of Leaf Vegetables-Brassicaceae, Leguminous Vegetables

於雲林縣水林鄉設置28 區合作田區，總面積7.27 公頃，擴大小菜蛾性費洛蒙使用面積，以達全面防治效果。以性費洛蒙誘集、監測之小菜蛾數換算成每公頃之誘殺數，每公頃估算之誘殺數為7502251237 隻，差異極大，顯示農民之管理模式差異極大。採收前共採樣22 樣品測定農藥殘留量，發現農民多混合多種農藥，其中有15 樣品未測出殘留量(ND)，6 樣品使用未登記藥劑，但多為有安全容許量而無使用方法之藥劑。利用空心圓錐型噴嘴以側噴方式噴施蘇力菌防治小菜蛾可發揮極佳之防治效果，且殘留檢測結果，施藥後第二天青花菜菜球之農藥殘留量已低於安全容許量，可知施藥方法可影響農藥殘留量。於雲林縣水林鄉建立五區豇豆試驗田區並完成二期作之管理模式，於信義鄉設置二菜豆試驗田區。於種植前進行土壤處理，種植後懸掛黃色及藍色粘板防除小型昆蟲，試用蘇力菌防除豆莢螟；建立腐霉病、萎凋病及立枯病三種土壤傳播性病害之共同防治策略，並建立蟲害整合防除策略。於採收期採樣進行農藥殘留量檢測，檢測結果分別為未檢出 (ND)、安全與使用未登記藥劑三種樣態；針對違規使用藥劑部份已詳細說明違規原因，並加以輔導。利用矽藻素撒施於土壤傳播性病害罹病植株周圍可有效抑制病害之擴展。 The main purpose of this project is to establish the health management strategies of leaf vegetable species, leguminous vegetables (including peas, beans, etc.). The strategies built can be used to help farmers to produce the health, safety and high quality of vegetables, and then maintain the health of the consumer concerns. The ICM program of both cowpea and two Kidney bean was established and held for five and two fields in Yunlin County and Nantou County. Twenty eight location of Broccoli was conducted with the area of 7.27 acre totally. The Diamondback moth (Plutella xylostella) occurred seriously. The sex pheromones is used to decrease the population of the diamondback moth. The hollow cone nozzle was used to control the diamondback moth and showed the significant efficacy by spraying the pesticides from side direction of the plants

Integration Crop Management of Leafy Vegetables - Brassicaceae, Water Spinach

於2010年1月份至2012年6月間在西螺鎮農友之簡易設施中共進行22期作十字花科小葉菜類整合管理試驗，發現較為嚴重之害物為黃條葉蚤，經浸水處理、播種後懸掛黃色粘板，配合少量之化學農藥噴施，可有效抑制黃條葉蚤之為害。由於黃條葉蚤移動性強，故噴施化學農藥防治時，可先噴施田區周圍，再向內依序噴施，降低成蟲向田區外遷飛，防除效果較佳。2009年8月進行溫室栽種蕹菜之整合管理試驗，至2010年6月該田區出現明顯之根瘤線蟲為害，經連續二期作澆灌甲殼素，無法發揮抑制線蟲為害，之後於2011年2月於罹病田區之田埂種植芳香萬壽菊，亦未有效抑制線蟲為害，反而引發介殼蟲發生；2012年6月輪作莧菜，採收時未發現線蟲為害，且連續種植6期作之蕹菜亦未見根瘤線蟲發生，至於此抑制之作用機制仍須進一步探討。 The integrated management of cruciferous vegetables was conducted in the simple vinyl house of Xiluo district for a total of 22 test from January 2010 to June 2012. The study found that the most serious problem is the striped flea beetle infestation. The practice, soil soaking treatment before sowing, hanging yellow sticky boards after sowing, following by a small amount of the pesticide use, can effectively inhibit the striped flea beetle infestation. Because the strong mobility of the striped flea beetle, to spray the pesticides area around the field first, and then again within the sequence spraying to reduce adult migration outside the fields, can control them effectively. The integrated management of water spinach at the greenhouse was held August 2009 and the Meloidogyne damage appeared at June 2010. The infestation of nematode could not been effectively suppressed by supplying chitin to the infected soil for two consecutive cultivation, followed by the planting of Tagetes lemmonii in February 2011. The practice was given up for the reason that the stem was infected by scale insects Finally. The nematode infestation condition was not found after amaranth crop rotation in June 2012, and maintained no infestation for six continuous water spinach cultivation. The mechanism of this inhibition still need further study

The Integrated Management on Strawberry Production in Organic Farming System

本年度將於大湖地區設置有機草莓試驗田，基肥施用依緩效性堆肥與速效性有機肥比例分為6:1、6:2及6:4（w/w）等三種處理，在總氮素施用量為300公斤/公頃為原則下分次追肥，控制肥料變因對草莓的影響，有助於調整施肥策略，減少肥料施用。土壤孵育試驗選定3種不同特性之土壤，探討有機質肥料礦化過程對草莓生長及土壤品質的影響，目的在於篩選適合有機草莓栽培之有機質肥料。此外本年度持續進行有機育苗試驗，追蹤有機草莓之病蟲害發生狀況，並提出各種解決方法。We will conduct organic strawberry field test at Da-Hu district. The different basal fertilizer treatments have three mixed rates with slow and fast-release organic fertilizers at 6:1, 6:2, and 6:4(w/w) respectively. The apply fertilizer will be restricted at 300 kg-N/ha. The mineralization rate of different formlus will be conducted with different in laboratory incubation. This will support our concept to make a good formula of organic fertilizer that can be used for maintaining soil quality, quaratee good production and simplfying fertilization practice on strawberry cultivation. We also carry on organic seedling raising and pest control test to full fill the strawberry cultivation in organic farming system

Chrsanthemum Diseases and Contol in Taiwan

臺灣菊花已知有＋一種病害，其中以莖腐病、白锈病、黑斑病發斥最為嚴重，黑鋸病、萎凋病、炭疸病偶有猖獗情形，其他有記錄或觀察到者有根腐病、褐锈病、褐斑病、灰徽病、白絹病。以季節來分，高溫時以莖腐病、萎凋病及黑斑病發生嚴重，冬季氣溫較低時，以白锈病及黑銳病發生較嚴重。 育苗期之莖腐病可取河川砂為苗床，插穗用新阿蘇仁1,000倍液浸10分鐘後沾植保素一號（內含NAA0.1％及萬力0.1%)，可防治該病之發生，並育出發根良好之健全苗。白锈病可用锈克粉劑4,000倍及乳劑1,000倍防治。白锈病病菌在埔里及陽明山地區能越夏，但在田尾、北斗及嘉義地區無法越夏。黑斑病可用四氯丹粉劑500倍，鋅鋼四氯丹粉劑600倍，或與免賴得粉劑1,500倍或新阿蘇仁溶液1,000倍輸流使用，防治黑斑病時亦可同時防治炭疽病。 清園對病蟲害之發生有極顯著的影響，收穫後應行清園，避免病蟲害之滋長，對菊花病蟲害之有效防治，極為重要。 Among 11 chrysanthemum diseases known in Taiwan, basal stem rot (Rhizoclonia solani), white rust (Pflccinia horiana) and black leaf spot (Septoria chrysanthemella) occur severely. Black rust (Puccina chrysanthemi), wilt (Fusarjum oxysporuni f. sp. chrysanthemum) and anthracnose ( Colletotrichum chrysanthemi ) occur sporadically. While brown rust (Uredo autumnalis), brown leaf spot(Septoria chrysanthemi-indica), gray mold (Botrytis cinerea) and southern blight (Scierotium rolfsii) had been recorded or observed to occur occasionally. In high temperature period, basal stem rot, wilt, anthracnose and blaek leaf spot are severe, while in winter, white rust, black rust and gray mold occur. Seed bed composes of river sand with cuttings are protected, after immersed 10 mill in 6.5% Neo Asozin S solution(1,000 timesdilution), by plant protection powder No.1 ( 0.2% NAA and 0.1% Benlate in talc) to control basal stem rot and to promote abundant rootlets formation. White rust fungus could over-summer in Puli and Yanminshan areas but did not so in Tienwei, Peitou and Chiayi areas. White rust is controlled by Plantvax (4,000 times dilution and 1,000 times dilution, respectively, for 75% WP and 20% EC). Black leaf spot is controlled with 80% Difolatan WP 500 times dilution or 50% Zincofol WP at 600 times dilution alone or rotating with 50% Benlate WP 1500 times dilution or 6.5% Neo Asozin S at 1000 times dilution. Same spray also controls anthracnose. Cleaning of the harvested fields is most important to control diseases and insets for the nearby fields

The Integrated Management on Strawberry Production in Organic Farming System

有機農業是一世界風潮，以最主要精神是在國家規定之規範下進行之一生產系統，在此系統必須同時兼顧環境品質及生態平衡。本研究目的在建立台灣各草莓產區有機栽培標準化綜合管理模式，包括有機農業在草莓栽培上土壤品質標準、土壤改良資材、肥料資材、病蟲害防治資材之選擇及使用。 本計畫分年目標：95年度：本年度進行有機草莓栽培上土壤性質分析、肥料 資材及用量、覆蓋資材之影響、病蟲害防治建議。96年度：依95年度成果進一步修改使有機栽培標準化綜合管理模式更適合農民使用，並作田間示範觀摩。97年度：依已建立模式推廣所有種草莓地區。 本研究由一工作團隊進行：一、進行有機草莓苗之栽培需要選擇適當地區栽培以健全有機栽培制度之建立。此部分由吳添益、蔡政賢負責田間控管外其他肥料及病蟲害管理由團隊其他負責人提供建議並評估可行性。二、.草莓為旱作作物，前作作物選擇若能降低或克服某些草莓之病蟲害原，則更容易進行病蟲害防治，降低生物性或物理性防治病蟲害之花費成本。因此在草莓主要生產區(苗栗縣大湖)之有機耕種農田，種植前做以下各種研究。三、研究植物性材質(稻草)、塑膠布覆蓋方法對雜草防治之效果。農委會苗栗改良場吳添益副研究員負責此部分及田間管理。草莓有機栽培之水分管理配合有機液肥之隨灌溉水提供草莓追肥，此部分由農業試驗所農業化學組向為民副研究員負責。有機草莓田土壤性質診斷及養分變化研究部分由中興大學黃裕銘副教授負責。草莓栽培氣候條件之選擇相當重要，但是栽種地區土壤問題需要診斷出來並給予改良則對有機栽培之成敗有相當重要之關鍵性，因此需要作了解。另外不同耕作制度會影響土壤中微生物族群及土壤性質而影響作物養分在土壤中之轉變速率，因此需要研究了解，才能給予明確之施肥制度。四、草莓之生產期長，且一面開花一面成熟，其基肥追肥之施用材料及配方和其他作物不同，需要利用地區性較便宜資材以建立之，另外追肥是否需要有機液肥之補充及有機液肥之配方與製作亦相當重要，因此需要研究。五、在不同耕作制度、覆蓋及施肥狀況下，有機草莓之病蟲害發生狀況需要明確追蹤調查並提出各種解決方法，尤其草莓栽培最嚴重之灰霉病和紅蜘蛛之危害需要從田間管理及肥料選擇及土壤生態營造之綜合管理著手成功率最高，此部分由農委會農業藥物毒物試驗所楊秀珠研究員負責。六、研究時程及進度本研究95年度及96年度各研究主題不變但是更進一層分析所建議技術之穩定性，97及98年進行推廣及研討會分享研究成果給國內外研究人員、農民及消費大眾參考。 本研究.預期效益：一、建立有機草莓生產標準模式。二、提高有機草莓之品質及可靠性。三、提升觀光產業。四、提升農民收益。The organic farming has been dramatically increased world wide. It is a production system under governmental orders or laws, and enchanced the environemnetal quality, including soil, water, and ecosystem. The aim of this study is to establish an intergrated organic farming system for growing strawberry in Taiwan, including soil quality, soil amended materials, fertilizers, pest control materials, and speicific managing practices adopted for different regions. The aim will be approached by several years. In 2006, experiment will be conducted in Mioli, the most famous strawberry production county. The soil properties will be analysed and set an appropriate system including soil amendments, fertilizers, mulching materials, water control, and pest survey and controls. The system conducted in 2006 will be modified in 2007 if it is necessary, and a field demonstration will be hold to open to farmers and any publics. The models will be extended to other regions with suitable modification according to regional natural environments. This project will be conducted by a team of researchers from different institutes including many soil scientists and plant pathologists. The field managements will be charged by Mr. Tsai, Dr. Tsai, and Mr. Wu; the irrigation and water control is charged by Mr. Hsiang; The pest control is charged by Dr. Young and Dr. Chang; and the fertilizer is charged by Dr. Huang. The rice straw and PVC film are used as mulching materials for weeds control and decrease the gray mould. The harvest seasons is long and the flowering stages covering fruit ripening stages, so its fertilization system will be very different to other tree fruit system. Some results and benefits will be derived from this study: An appropriate integrated management system will be established for farmers to follow up with stable production and quality of strawberry in organic system. Which returns the farmers with good farm-gate price and profits and enhanced the sightseeing population for all regions

動物疫病診斷鑑定技術之研發與應用

1. The foot and mouth disease (FMD) eradication program has been an important policy in Taiwan, and it leads to be recognized as a FMD free country. At present, commercialized antibody detection kits mainly use the method of enzyme-linked immunosorbent assay (ELISA). In this project, we are planning to develop an antibody detection kit using immunodot method. The antigen used will be expressed protein of VP1, VP2, and 3AB genes of FMD virus. This newly developed antibody detection kit will be able to be used in the field without using expansive equipment. This development should be very helpful to the execution of the FMD eradication campaign in the future. 2. HPAI of the H5 subtype has imposed a great potential threat to both the poultry industry and humans in Taiwan. To effectively monitor and manage the disease, we have previously developed a rapid virus detection kit based on the monoclonal antibodies against the HA protein of avian influenza virus H5 subtype for the on-site operations of the front-line workers. To further test the sensitivity and specificity of the rapid detection kit with clinical samples, we will continue the collaboration with Animal Health Research Institute to perform the virus challenge assays with H5N2 (or H5N1) and H6 subtypes of AIV under Bio-Safety Level 3 environments. Scheduled works includes the clinical evaluation for the sensitivity, specificity of the rapid virus detection kits, as well as training and extension for the use of the kit. It is expected that the rapid virus detection kit for avian influenza virus H5 subtype will serve as a tool for the efficient, rapid and initial on-site screening of the diseases, and facilitate the effective control of the viruses to ensure the sustainable poultry industry in Taiwan. 3. Enterotoxigenic E. coli (ETEC), Salmonella Typhimurium, S. Choleraesuis, Clostridium perfringens etc., are major pathogens for porcine diarrhea cases. On the other hand, Staphylococcus aureus, some Streptococcus spp. and Corynebacterium bovis may cause mastitis for bovine. In comparison with conventional PCR, Real-time PCR has recently become a popular method. It does not require the step of gel electrophoresis, is time saving, more sensitive and causes non-cross reaction. In this study, by use of SYBR Green or TaqMan Probe, we shall design Real-time PCR method for the specific detection of Salmonella Typhimurium, S. Choleraesuis, ETEC and Clostridium perfringens. Meanwhile, Real-time PCR quantation for these pathogenic bacteria will be tried since pathogenicity of bacteria is highly correlated with its cell number. Finally, we shall also try the possibility to develop a multiplex Real-time PCR system for the simultaneous detection of these diarrheagenic bacteria. Through this project, we expect to establish a rapid Real-Time PCR method which allows us to trace the cause of diarrhea and infections for porcine and thus prevents the spreading of infection. Since S. Typhimurium, S. Choleraesuis, ETEC, and C. perfringens are common pathogens for human and porcine, this method can be usd to trace the contamination sources for both human and porcine infections. Finally, we shall also try to develop a multiplex Real-Time PCR system and use it for the simultaneous detection of these pathogenic bacteria. Also, patent application and technique transfer to bioindustry will be attempted. 4. Hepatitis E is a non-A, non-B hepatitis and is transmitted by the fecal-oral route. Contamination of drinking water has become the main reason to cause outbreaks of hepatitis E in several developing countries. It is an important disease with public health concern. The mortality rate of hepatitis E is generally low, about 1% only; however, it can be more serious in一、口蹄疫抗體快速檢測試劑之研發：口蹄疫的撲滅計畫已被列為我國的重大防疫政策，而目前實驗室中對於口蹄疫的診斷方法有螢光抗體檢測法、病毒分離、RT-PCR及免疫吸附法(ELISA)等方法，以上檢測方法所需時間長，且需要特殊儀器，無法於現場使用，因此需開發一方便快速的檢驗方法，以便在現場快速檢測並立即摘除帶原者。本實驗將針對口蹄疫病毒的VP1、VP2及3AB基因，來進行原核系統的表現與純化，以開發出一種較現有檢測方法更具有經濟效益及特異性、敏感性、簡便而快速的口蹄疫抗體檢測方法以提供臨床檢測之用。本計畫將與國內知名且具高度研發生產及行銷能力佳之生物技術公司進行產學合作，利用他們的免疫墨渍檢測法平台開發出快速現場檢測口蹄疫抗體的診斷試劑套組，並期待於未來予以商品化，未來將有助於協助監測口蹄疫疫苗注射狀況，及進行現場口蹄疫病毒感染豬隻檢測與清除。 二、流感病毒H5亞型快速檢測試劑套組之實測與應用：H5亞型之高病原性家禽流行性感冒(HPAI)對台灣養禽業具極大之潛在危險，並有人畜共通疾病之隱憂。為求有效地監控此病毒，本計劃上年度已初步完成禽流感病毒H5亞型病毒快速檢測試劑套組。本年度由中興大學與家畜衛生試驗所合作，繼續以H5N2 (或H5N1)與H6N1亞型禽流感病毒進行攻毒試驗。預期將可利用禽流感病毒時體樣本完成禽流感病毒快速檢測套組之實體樣本測試與評估，並進行人員訓練與推廣工作，未來可供第一線工作人員作為現場快速病毒篩檢之基本工具，防堵病毒入侵與擴散，確保我國養禽產業之永續經營。 三、開發多套式即時聚合酶鏈鎖反應快速檢測豬下痢病原菌及牛乳房炎病原菌之技術及其應用：豬下痢及腸炎之病原菌，包括腸毒型大腸桿菌(ETEC)、沙門氏菌(如S. Typhimurium、S. Choleraesuis)、腸道螺旋體、Clostridium perfringens…等，另一方面，造成牛乳房炎的病原菌，則包括葡萄球菌、鏈球菌、棒狀桿菌、大腸菌…等。與傳統PCR比較，Real-Time PCR 不需電泳操作且靈敏度提高，受檢測菌數不必增殖，故能大大減少縮短檢測時間，並避免偽陽性的發生。因此本研究擬利用Real-Time PCR配合SYBR Green染劑或TaqMan probe方法，設計沙門氏菌屬如S. Typhimurium、S. Choleraesuis及ETEC、產氣莢膜桿菌專一性引子組及專一性TaqMan探針，用以快速鑑定禽畜類重要之感染性病原菌，由於致病性與菌量亦有相關性；因此發展同時檢測多種病原菌並定量之快速方法於防治感染是必要的。本計畫擬分年開發多套式Real-Time PCR，針對上述細菌中S. Typhimurium, S. Choleraesuis、ETEC（LT及ST）及Clostridium perfringens等設計專一性引子組及TaqMan探針，以Real-Time PCR檢測定量豬隻感染之重要病原菌。本研究預計可建立一套快速檢測豬隻下痢病原菌的即時PCR套組（Real Time PCR），了解豬隻下痢之病因，達到傳染性胃腸炎之快速防制、避免擴散等目的。S. Typhimurium、S. Choleraesuis、ETEC（含heat-liable toxin, LT, 及heat stable toxin, ST）及產氣莢膜桿菌為人畜共通病原菌，本方法亦可應用於人類、豬隻感染上述病原菌之病因，對相關傳染途徑予以控制。並可設計一套多重檢測之 Real-Time PCR套組，用於多種細菌之同時檢測。相關PCR套組可申請專利移轉生物技術產業使用。 四、人畜共通傳染病原: 豬E型肝炎病毒快速檢測技術之開發：E型肝炎是經口糞感染的非A及非B型肝炎，在開發中國家因水源遭受污染常有爆發性流行，為一重要的公共衛生疾病。E型肝炎感染者一般死亡率約在1%左右，但在孕婦，尤其第三期妊娠感染者，死亡率可達20%。E型肝炎病毒在豬隻身上亦被發現，在日本曾有因直接生食野鹿肉或野豬肝臟而導致發病死亡的病例；此病原在豬隻飼養、屠宰及獸醫師等相關人員有明顯高血清抗體陽性率。台灣豬E型肝炎病毒感染相關情形雖有部分報告，但相關的研究及疫情掌握仍有不足，為因應未來此一人畜共通傳染病及畜產品產製環境之人畜安全檢測需要，本計劃擬建立及開發豬E型肝炎病毒之快速診斷技術及試劑。 五、豬流感病毒單源抗體製備、特性分析與診斷套組開發：臺灣豬群中流行的A型流行性感冒病毒，則除了古典豬型H1N1亞型及似人型H3N2亞型之外，近期也偵測出H1N2亞型及H3N1亞型等二種新興病毒變異株。鑑於現有檢驗診斷技術方法已不足以有效偵測新型