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)

2 (Oncorhynchus mykiss) (Salmo trutta) (Thymallus thymallus) (Coregonus sp.) (Esox lucius) (Scophthalmus maximus) (O. 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.)

3 R. Winton )22 VHSV (1~5 C ) (15~18 C ) VHS (5) VHS ( 2) ( 3) ( ) 2 VHS (P.

4 Ghittino ) 3 VHS ( Vol. 15, No. 4 "What should do?" )(6) 2. (1) 10 48 23 (2) MS222(Tricaine.

5 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.

6 (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.

7 (1) VHSV :BF-2 EPC ( 7) : (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 ( )

8 (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 ( ) 500 L 2 -20 C 20 (24 ) 25 C 24 80% BF-2 EPC ( 6 2 2 ))

9 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 ( )

10 1 PBS-Tween 4 37 C 1 FITC (2 ) 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.