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海洋資源之生物科技應用 ( 二 )

海洋資源之生物科技應用 ( 二 ). negative very light light medium severe. Environmental factors Broodstock Nauplius to PL Juvenile Feed. indicates the condition should be safe from WSSV epidemic indicates there is a potential risk of WSSV epidemic, extra care needs to be taken

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海洋資源之生物科技應用 ( 二 )

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  1. 海洋資源之生物科技應用(二)

  2. negative very light light medium severe Environmental factors Broodstock Nauplius to PL Juvenile Feed indicates the condition should be safe from WSSV epidemic indicates there is a potential risk of WSSV epidemic, extra care needs to be taken indicates WSSV epidemic may occur any moment, stop culturing is recommended Infected virus titer High Low WSSV Index Detection limit

  3. Penaeus monodon Portunus sanguinolentus Portunus pelagicus Artemia Peneaeus japonicus Scylla serrata Penaeus semisulcatus Calappa philarigus Family: Ephydridae Charybdis feriatus Penaeus penicillatus Charybdis natator Helice tridens Schmackeria dubia Metapenaeus ensis Macrobranchium rosenbergii Exopalaemon orientis

  4. Possible sources and transmission pathways of WSSV within a closed system Grow-out pond Hatchery Vertical transmission (transovarial transmission transovum transmission) Cannibalism Decomposition of dead shrimp Female brooders Cultured Shrimp Stocking Ingestion Immersion Offspring Ingestion Immersion Decomposition of dead shrimp Reservoir host Water Food Cannibalism Spermatophore Reservoir host Water Food Ingestion Immersion mating Decomposition of dead shrimp Male brooders potential source possible pathway

  5. WSSV from different species of shrimps and crabs are closely related if not identical

  6. Is WSSV a natural virus commonly present in wild populations of crustaceans?

  7. Outbreaks of WSS 1992 China, Taiwan 1993 Japan, Thailand, Korea 1995 USA

  8. MT WSSV ? MBV ? WSSV Annual productions of cultured Penaeus monodon in Taiwan

  9. Detection of WSSV in Callinetes sapidus & Artemia sp.

  10. Virus like particles found in the shell gland of Artemia (Criel G. 1980)

  11. 2-step WSSV diagnostic PCR Number of Source Detection rate C. sapidus tested 1-step positive 2-step positive New York 110 0 32 29.1% Texas 51 0 13 25.5% New Jersey 45 0 11 24.4% Total 206 0 56 27.2% 2-step WSSV diagnostic PCR screening of C. sapidus WSSV detection rates inwild populations of C. sapidus collected from US coastal waters

  12. 2-step WSSV PCR screening of Artemia sp. WSSV PCR screening of Artemia using five primer sets derived from the WSSV pms 146 genomic fragment

  13. Days post-hatching 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Day-0 group Day-1 group Day-2 group Day-3 group Day-4 group Day-5 group Day-6 group Day-7 group Control : pre-challenge period WSSV testing by 2-step PCR :challenge by immersion : post-challenge period Protocol of the experimental challenge of Artemia with WSSV

  14. day-1 day-2 day-0 IQ2000 pms 146 day-3 day-4 day-5 IQ2000 pms 146 day-6 day-7 control IQ2000 pms 146 Experimental challenge of Artemia with WSSV Infection rates of WSSV challenged Artemia

  15. nd: no difference – : reaction not performed

  16. Restriction fragments of WSSV genome M1 HindIII XhoI SalI M2 XhoI SalI M1

  17. 10_11_BAC.clones.jpg

  18. 09_27_Computer.prog.jpg

  19. 10_15_DNA.microarrays .jpg

  20. Hours postinfection Functional genes in cluster A: vp53A, vp73 3 % Functional genes in cluster B: vp11, vp12B, vp13B, vp15, vp26, vp35, vp36B, vp51A, vp51B, vp51C, vp53C, dUTPase,tk-tmk, pk2 20 % Functional genes in cluster C: 15 % vp75, ts, rr1, rr2, pk1 11 % Functional genes in cluster E: vp110 12 % Functional genes in cluster F: vp12A, vp19, vp22, vp24, vp28, vp31, vp32, vp36A, vp38B, vp39A, vp39B, vp41A, vp41B, vp55, vp60B, vp95, vp136A, vp136B, vp180, vp186, dnapol 33 % 0 2 4 8 12 18 24 6 A B C D E AF440570 F

  21. WSSV M (kDa) 1 212 2 170 3 4 116 5 6 7 97 8 9 76 10 11 12 66 13 14 15 53 16 17 45 18 19 20 21 22 23 24 30 25 26 27 28 29 • 20.1 30 31 • 14.4 32 33 34

  22. Localization of WSSV VP28 N I N I N E E E I N 100 nm 100 nm Envelope Tail-like projection Integument Nucleocapsid

  23. Localization of WSSV VP26 Nu Cy 250 nm 200 nm

  24. WSSV M (kDa) 1 212 2 170 3 4 116 5 6 7 97 8 9 76 10 11 12 66 13 14 15 53 16 17 45 18 19 20 21 22 23 24 30 25 26 27 28 29 • 20.1 30 31 • 14.4 32 33 34 Localization of WSSV ORF419

  25. Viruses prevalence in shrimps captured from southern Taiwan coast waters

  26. What else left for us to do in WSSV Establishment of WSSV susceptive cell lines Screening of WSSV pathogenicity related genes Studying on WSSV morphologenesis Development of anti-WSSV agents or strategies Investigating the usages of WSSV genes

  27. pIZΔIE/WSSV126-2k/V5-EGFP-His pIZΔIE/WSSV126-1k/V5-EGFP-His pIZ/V5-EGFP-His BF EGFP pIZΔIE/WSSV126 rev/V5-EGFP-His pIZΔIE/WSSV126-2k/V5-EGFP-His pIZΔIE/WSSVdnapol/V5-EGFP-His pIZΔIE/WSSV126-1k/V5-EGFP-His pIZΔIE/V5-EGFP-His pIZ/V5-EGFP-His anti-EGFP anti--actin

  28. 水產養殖生技產業之競爭力(SWOT)分析 1. 產業優勢（Strength） (1) 我國水產繁養殖技術享譽全球，所累積之經驗及產業基礎均具國際競爭之利基。 (2) 水產相關學術研究機構培育眾多優秀水產生物技術人才，並擁有豐富之研究成果。 (3) 國內擁有龐大資金，透過政府大力投資將可帶動生技產業迅速發展。 2. 產業弱勢（Weakness） (1) 水產生物技術研究領域過於分散，未能針對具利基及潛力之領域進行整合，相關研發成果尚未與產業結合及產業化。 (2) 產業界對水產生物技術之發展潛力認知尚嫌不足，缺乏投入之勇氣。 (3) 水產生物技術產業的資訊極端不足，無法進行有效規劃。 3. 產業機會（Opportunity） (1) 水產生技產品附加價值高，產品壽命及價值鏈長。 (2) 生物技術產業已列為新興重要策略性產業發展推動項目。 (3) 熱帶及亞熱帶國家養殖產業發展迅速，可開發溫水魚養殖相關生技用品。 4. 產業威脅（Threat） (1) 鄰近國家（韓國、日本及新加坡）政府大力推展生物技術產業，我國投資環境優勢將逐漸消失。 (2) 國內生產成本較高，造成競爭力低落。 (3) 對岸營造優良投資環境，關鍵技術及產業資訊外流速度增快。

  29. Commercialize Research and Development After Service and Bio-solution Marketing What do a biotechnology company do?

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