ウイルス性出血性敗血症 VHS：Viral Hemorrhagic …

1 21 VHS Viral Hemorrhagic Septicemia1. (1) Viral Hemorrhagic Septicemia (VHS) (Egtved) (INuL) VHS ( 1) VHSV 3 VHSV 3 VHSV 4 (2) VHS VHSV 1996 VHS (3) (Oncorhynchus mykiss) (Salmo trutta) (Thymallus thymallus) (Coregonus sp.) (Esox lucius) (Scophthalmus maximus) (O.)

2 Tshawytscha) (O. kisutch) (Clupea pallasi) (Gadus macrocephalus) (G. morhua) 10 VHSV VHSV (Paralichthys olivaceus) (Trachurus japonicus) (Ammodytes personatus) (Sebastes inermis) (S. oblongus) (Pagrus major) (4) VHSV VHSV VHSV VHSV 1 VHS (J. R. Winton )22 VHSV (1~5 C ) (15~18 C ) VHS (5) VHS ( 2) ( 3) ( ) 2 VHS (P.

3 Ghittino ) 3 VHS ( Vol. 15, No. 4 "What should do?" )(6) 2. (1) 10 48 23 (2) MS222(Tricaine; 3-aminobenzoic acid ethyl ester methanesulfonate) 70% ( 48 ) (3) (a) 4cm (b) 4cm~6cm (c) 6cm ( )(a) (b) 1 1 5 1 1mL (4) / 4 C 2( 5 ) 2.

4 (MEM) (HBSS) 1000 g/mL 800IU/mL 800 g/mL 400IU/mL 12 5~10% pH ~ 2M (a) (0~4 C ) (b) 15 C 2~4 4 C (c) (d) 1:10 24 (e) 2000 g 15 (f) ( ) 3. (1) VHSV :BF-2 EPC ( 7).

5 (FCS)10% (MEM) ( 100IU/mL 100 g/mL) (a) 24 BF-2 EPC 20 C 24 (b) 10 3 BF-2 EPC (100 L/2cm2 ) (c) 10~15 C ~1 FCS 2% ( ) 24 1mL/ 15 C (a) 40~100 7~10 (b) pH ( ) 2M ( ) (c) (CPE)( 5) ( ) 4 VHS EPC ( 5 VHS EPC CPE (T R Meyers ) 25(d) CPE 7 CPE (a) (b) 4 C 15 2,000 g (c) BF-2 EPC (d) CPE (2) Bio-X BIO-FLUO VHS IP5B11(Bio K 006) (a) 100mL 1 PBS 900mL 10 (b) 25mL 1 (c) VHS 1 500 L 1 -20 C 10 20 (d) FITC ( ))

6 500 L 2 -20 C 20 (24 ) 25 C 24 80% BF-2 EPC ( 6 2 2 ) 10 VHSV 5,000~10,000 PFU/mL 15 C CPE 500 L 4 C 20 1mL 200 L 1 1 1mL 200 L 2 1 1mL ( ) VHSV 26 (3) (2) ~ PBS( ) 1 (-20 C ) 30% 70%( / ) 3 -20 C 2cm2 500 L 15 30 -20 C Tween80 PBS (PBS-Tween ) VHSV ( ) PBS-Tween 4 37 C ( ) 1 PBS-Tween 4 37 C 1 FITC (2 )

7 PBS-Tween 4 ( ) (4) RT-PCR (a) ( ) RNA(b) RT-PCR (c) Forward primer : 5'-GGG GAC CCC AGA CTG T-3'Reverse primer : 5'-TCT CTG TCA CCT TGA TCC-3' :811bp (a) RNA (b) RNA PCR 50 C 30 94 C 2 94 C 30 52 C 30 68 C 1 35 68 C 7 (c) PCR DNA 1% (d) 811bp 274.

8 Manual of Diagnostic Tests for Aquatic Animals 2006, CHAPTER , OIE ( )