(Pelteobagrus fulvidraco) FJ766525), (Edwardsiella ictaluri)

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1 41 1 Vol.41, No OCEANOLOGIA ET LIMNOLOGIA SINICA Jan., 2010 (Pelteobagrus fulvidraco) (Edwardsiella ictaluri) * ( ) 2 (CHNYC001 CHNYC002),,, 16S rdna (GenBank FJ FJ766525), (Edwardsiella ictaluri) 20 30, ph 7.0, 2%,,,,,,,,,, S (Pelteobagrus fulvidraco),, (Siluriformes) 鲿 (Bagridae) (Pelteobagrus) (, 1994),,,,,,,,,,, (, 2008), (2008) (Enteric Septicemia of Catfish, ESC), ESC (Edwardsiella ictaluri), ; (2008) (Edwardsiella tarda) , 40% 50%, 90% 2 (CHNYC001 CHNYC002),, 16S rdna,,, (Pelteobagrus *, IRTO848 ;, 08ZA082,,, gengyisicau@126.com :,, kywang@sicau.edu.cn : , :

2 62 41 fulvidraco), ;, ( )g (BHI), TSA BHI ; ; DNA PCR Takara , 1.2.2, BHI, h,,,,, Morrison (1994) ( 2007), BHI 28 48h,, McFarland 0.5, 20,, 23 25,,, CFU/ml, 0.1ml,, 14, CFU/ml 1h,, BHI TSA BHI, h,,, (, 2001) S rdna (1) PCR DNA : BHI, 28 36h, 4ml, 10000r/min 2min, DNA DNA PCR (2) 16S rdna : 1 16S rdna (, 2006), : 5 -AGAGTTTGATCCTGGCTC AG-3 5 -TACGGCTACCTTGTTACGAC-3 PCR : 94 5min; 94 1min, 54 1min, 72 2min, 30 ; 72 10min PCR, Takara (3) : 16S rdna GenBank Blast,, (2007), ClustalX 1.8, MEGA 4.1 Neighbor- Joining, , BIH, 28 24h, CFU/ml BIH,, 28 24h, ,,,,, 3 5,,,,,,,,,,,,,,,,,,, BHIA, 28 48h, CHNYC001 CHNYC002 BHIA, 28 48h, mm,,,

3 1 : (Pelteobagrus fulvidraco) (Edwardsiella ictaluri) CHNYC001 CHNYC , 95.0%(19/20) 100.0%(20/20),, CHNYC001 CHNYC002 3, 5, 7 12, 65%(13/20),, 84.6%(11/13), 15, 15.4%(2/13), BHI,,,, 2.4 CHNYC001 CHNYC002 G-,,,,, ( )μm ( )μm, TSA,, 48h 0.1mm,,, BHI 2.5 CHNYC001 CHNYC002 ( 1), 15 40, 表 1 菌株 CHNYC001 与 CHNYC002 的理化特性 Tab.1 Biochemical and physiological characteristics of CHNYC001 and CHNYC002 CHNYC001 CHNYC002 CHNYC001 CHNYC V-P 45 NaCl 0% + + H 2 S 0.5% % D- 1.5% + + D % ph D L L / (O/F) F F D DNA : +,, F, O

4 , 2%, ph , ph 7.0;, D-, DNA V-P,, H 2 S S rdna PCR PCR CHNYC001 CHNYC002 16S rdna 1500bp, 1506bp 1513bp, GenBank FJ FJ GenBank Blast, 16S rdna MEGA 4.1 ( 1), CHNYC001 CHNYC002 鮰 (Edwardsiella ictaluri), 99.6% 100.0%, 鮰 (Edwardsiella ictaluri) 1 16S rdna Fig.1 Phylogenetic tree in the alignment of the 16S rdna sequence : 16S rdna GenBank , CHNYC001 CHNYC002, G ;, CHNYC001, CHNYC002, CHNYC001, CHNYC ,,, (2008) (Edwardsiella tarda); (2008) (Enteric Septicemia of Catfish, ESC),, 2 CHNYC001 CHNYC002,,,, 16S rdna, CHNYC001 CHNYC , (Hawke, 1979; Hawke et al, 1981), ( ) 鮰 (Crumlish et al, 2002; Woo, 1999;, 2008), 鮰 (white catfish)(plumb et al, 1983) (sea bass)(blanch et al, 1990) (walking catfish)(kasornchandra et al, 1987) (brown bullhead)(plumb et al, 1983) (Japanese eel)(chung et al, 1983) (Bengal danio)(waltman et al, 1985) (green knife fish)(waltman et al, 1985) (brown bullhead)(iwanowicz et al, 2006) (striped catfish)(yuasa et al, 2003) (Tra catfish)(crumlish

5 1 : (Pelteobagrus fulvidraco) (Edwardsiella ictaluri) 65 表 2 菌株 CHNYC001 与 CHNYC002 的药物敏感性 Tab.2 The sensitivity of CHNYC001 and CHNYC002 to antibiotics (mm) (μg/ ) CHNYC001 CHNYC002 CHNYC001 CHNYC002 G Penicilin R R Ampicillin R R Amoxicillin R S Cefalexin MS MS Cefazolin MS MS Norfloxacin S R Ciprofloxacin S S Enrofloxacin S MS Lomefloxacin S S Norfloxacin S R Cidomycin MS S Kanamycin R MS Amikacin MS S Tenemycin R S Fradiomycin S S Tenemycin R R Ambramycin S S Deoxycycline S S Minocycline MS MS Florfenicol S S Sulfamethoxazole R R Azithromycin R R Medecamycin R R Acetylspiramycin R R : R, S, MS et al, 2002) (rainbow trout)(oktay, 2004),,, 3.2,,,,,,,,,,,,,, 鮰,,,,,, (Newton et al, 1989; Nusbaum et al, 1996, 2002);,, (Morrison et al, 1994; Newton et al, 1989),,,,,

6 , (1) :, E Zn (Paripatananont et al, 1995; Wise et al, 1993);, (Wise et al, 2008),, (2) :, (Noga, 1995), (Crumlish et al, 2002; Haesebrouck et al, 2008),, (Anissa et al, 2003; Gaunt et al, 2006; Haesebrouck et al, 2008;, 2007),,,,,, :, ,,, , 31(4): ,,, (Pelteobagrus fulvidraco) (Edwardsiella tarda)., 39(5): ,,, , 39(2): ,, :, , , 1: 56 57,,, RH2., 31(4): ,,, , 46(4): ,,, , 27(4): Anissa M P, Gaunt T, Santucci R et al, In vitro evaluation of the susceptibility of Edwardsiella ictaluri, etiological agent of enteric septicemia in channel catfish, Ictalurus punctatus, to florfenicol. Journal of Veterinary Diagnostic Investigation, 15(5): Blanch A R, Pinto R M, Jofre J T et al, Isolation and characterization of an Edwardsiella sp. strain, causative agent of mortalities in sea bass (Dicentrarchus labrax). Aquaculture, 88(3 4): Chung H J, Kou G H, Edwardsiella ictaluri isolated from cultured eel in Taiwan Fish Disease Research, 5(1): Crumlish M, Dung T T, Turnbull J F et al, Identification of Edwardsiella ictaluri from diseased freshwater catfish, Pangasius hypophthalmus (Sauvage), cultured in the Mekong Delta, Vietnam. Journal of Fish Diseases, 25(12): Gaunt P S, McGinnis A L, Santucci T D et al, Field efficacy of florfenicol for control of mortality in channel catfish, Ictalurus punctatus (Rafinesque), caused by infection with Edwardsiella ictaluri. Journal of the World Aquaculture Society, 37(1): 1 19 Haesebrouck T D, Sorgeloos F N A, Baele P et al, Antimicrobial susceptibility pattern of Edwardsiella ictaluri isolates from natural outbreaks of bacillary necrosis of Pangasianodon hypophthalmus in Vietnam. Microbial Drug Resistance, 14(4): Hawke J P, A bacterium associated with disease of pond cultured channel catfish. Journal of the Fisheries Research Board of Canada, 36(12): Hawke J P, McWhorter A C, Steigerwalt A C et al, Edwardsiella ictaluri sp. nov. the causative agent of enteric septicemia of catfish. International Journal of Systematic Bacteriology, 31(4): Iwanowicz L R, Griffin A R, Cartwright D D et al, Mortality and pathology in brown bullheads Amieurus nebulosus associated with a spontaneous Edwardsiella ictaluri outbreak under tank culture conditions. Diseases of Aquatic Organisms, 70(3): Kasornchandra J, Rogers W A, Pulmb J A et al, Edwardsiella ictaluri from walking catfish. Thailand: Journal of Fish Diseases, 10(2): Morrison E E, Plumb J A, Olfactory organ of channel catfish as a site of experimental Edwardsiella ictaluri infection. Journal of Aquatic Animal Health, 6(2): Newton J C, Wolfe L G, Grizzle J M et al, Pathology of experimental enteric septicemia in channel catfish Ictalurus punctatus following immersion exposure to Edwardsiella ctaluri. Journal of Fish Diseases, 12(5): Noga E J, Fish Disease Diagnosis and Treatment. Mosby-Year Book, Inc, Nusbaum K E, Morrison E E, Entry of 35S-labeled Ed-

7 1 : (Pelteobagrus fulvidraco) (Edwardsiella ictaluri) 67 wardsiella ictaluri into channel catfish. Journal of Aquatic Animal Health, 8(2): Nusbaum K E, Morrison E E, Edwardsiella ictaluri bacteraemia elicits shedding of Aeromonas hydrophila complex in latently infected channel catfish, Ictalurus punctatus (Rafinesque). Journal of Fish Diseases, 25(6): Oktay K, Edwardsiella ictaluri infection in rainbow trout (Oncorhynchus mykiss). Turkish Journal of Veterinary and Animal Sciences, 28(6): Paripatananont T, Lovell R T, Responses of channel catfish fed organic and inorganic sources of zinc to Edwardsiella ictaluri challenge. Journal of Aquatic Animal Health, 7(2): Plumb J A, Sanchez D J, Susceptibility of five species of fish to Edwardsiella ictaluri. Journal of Fish Diseases, 6(3): Waltman W D, Shotts E B, Blazer V S, Recovery of Edwardsiella ictaluri from Danio (Danio devario). Aquaculture, 46(11): Wise D J, Greenway T, Li M H et al, Effects of variable periods of food deprivation on the development of enteric septicemia in channel catfish. Journal of Aquatic Animal Health, 20(1): Wise D J, Tomasso J R, Schwedler T E et al, Effect of vitamin E on immune response of channel catfish to Edwardsiella ictaluri. Journal of Aquatic Animal Health, 5(3): Woo P T K, Fish diseases and disorders (volume 3 Viral, Bacterial and fungal infections). CABI publishing, New York, Yuasa K, Edy Barkat Kholidin Panigoro N, Hatai K et al, First isolation of Edwardsiella ictaluri from cultured striped catfish Pangasius hypophthalmus in Indonesia. Fish Pathology, 38(4): ISOLATION, IDENTIFICATION AND BIOLOGICAL CHARACTERIZATION OF EDWARDSIELLA ICTALURI FROM CULTURED YELLOW CATFISH PELTEOBAGRUS FULVIDRACO GENG Yi, WANG Kai-Yu, FAN Fang-Ling, CHEN De-Fang, HUANG Jin-Lu (Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Ya an, ) Abstract Bacterial strains CHNYC001 and CHNYC002 were isolated from cultured yellow catfish Pelteobagrus fulvidraco with red-head disease in Meishan and Xinjin counties, Sichuan Province. The strains were proved to be the pathogens of the disease in our challenging experiments by immersion and intraperitoneal injection. The isolated strains were identified as Edwardsiella ctaluri in morphology, physiology and biochemistry, as well as 16S rdna sequence (Accessible Number in GenBank: FJ766524, FJ766525). The optimum temperature and ph for the strains growth was and 7.0. The strains grew normally at <2% in salinity of the culture medium. The results also show that the strains were sensitive to florfenicol, doxycycline, and lomefloxacin, but to acetylspiramycin, sulfamethoxazole, and medemycin. Clinical and pathological study revealed two types of the Edwardsiella ictaluri-induced disease in yellow catfish: acute and chronic. The acute type featured mainly septicemia, necrosis, haemorrhage, and inflammatory reaction in multi-organs and tissues, while the chronic one showed hemorrhage and ulcer on head, instead. Key words Pelteobagrus fulvidraco, Red-head disease, Edwardsiella ictaluri, Isolation, Identification, Biological characteristics