ISOLATION AND ID ENTIFICATION OF PORCINE REPROD UCTIVE AND RESPIRATORY SY ND ROME ( PRRS) VIRUS IN CHINA

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Aldous Wade
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1 1997,20 (3) : Journal of N anjing A gricult ural U niversity Ξ (, ) 7, Marc J1 J2 J3 (56,1 h), nm, nm, Marc2145,52 22 (BUDR), 3 ( PRRSV), PRRSV ; ; S ISOLATION AND ID ENTIFICATION OF PORCINE REPROD UCTIVE AND RESPIRATORY SY ND ROME ( PRRS) VIRUS IN CHINA Jiang Ping, Jian Zhongyou, Ma Zhiyong, Cai Jiali and Cai Baoxiang (Animal Infectious Disease Lab, Nanjing Agric U niv, Nanjing ) ABSTRACT On base of the epizootiology of seven pig herds with porcine reproductive and respiratory syndrome (PRRS) outbreaks in China, three virus strains were isolated from aborted fetues in the herds on Marc2145 cell culture. The three isolates, designated as J1, J2 and J3, could cause cytopathic effects (CPE), which could not be inhibited by BUDR. The causal virus did not resist treatments with chloroform,p H 5. 0, p H 8. 0 and 56, and decreased significantly in infectivity when exposed to 0. 2 % formalin for 2 h at 37. The diameter of the virus detected in cytoplasm of Marc2145 cells was nm, but in negatively stained electron micrographs was nm. They showed serological relationship to the American serotype of PRRS virus. The results confirmed that PRRS virus was present in China. Key words porcine reproductive and respiratory syndrome virus ; isolation ; identification ( PRRS) ( PRRSV), [1 ] 1987, [2 8, ] [9, ] PRRSV, Ξ ( ) (BE 96404) :

2 3 : ,,, EL ISA PRRSV 41, (, Herd Chek, IDEXX) PRRSV, ( ) ( ) 113 Marc2145 Kim [10 ] Marc2145 ( ) 8 % Eagle MEM,p H [2 Collins ] 3 5 ( ) Eagle MEM, 3,1000 r/ min 20 min ;, - 50, h Marc2145,1 ml/ ; h = 4 % Eagle MEM, 6 9 d (CPE), CPE 70 %, CPE, Marc CPE Marc2145, h, 10 min, PBS 3, 1 20 PRRSV (, ) 37 1 h, PBS 3 Marc PRRSV ( Illinois Jin ),37 1 h TCID 50, Marc2145, h, ,6 000 r/ min 3 min,, r/ min 2 h (55. 2 Ti Bechman, L8255M), PBS, 200,

84 20 2 211 1996 3, 7, 1995 12 1996 3 40 41 84 % 90 %, 2213 % 3714 %, 1217 % 5915 %, 012 %,, 63 % 65 %,2 3, ;, 50 %,,,,, PRRSV EL ISA 41, 6 24 PRRSV 6 PRRSV 7317 %, 3815 %, 50 %,1 PRRSV 212 Marc2145

3 , 7, % 90 %, 2213 % 3714 %, 1217 % 5915 %, 012 %,, 63 % 65 %,2 3, ;, 50 %,,,,, PRRSV EL ISA 41, 6 24 PRRSV 6 PRRSV 7317 %, 3815 %, 50 %,1 PRRSV 212 Marc2145 4, 3 2 3, h CPE( 1) 6,CPE,,, TCID / ml 1 Marc2145 Fig. 1 A Cytopathic effect (CPE) caused by isolate on Marc2145 cell culture 213 A1 infected cells ; B1 normal cells 3, Marc h,

3 : 85 CPE, Reed2Muench TCID 50 1 3, (56 ), (p H 5. 0), (p H 8.

Treatment methods TCID 50 / ml (log10) J1 J2 J3 ( = 5 %) 4,24 h 0 0 0 4, 1 h 5. 83 5. 83 5. 5 Untreated 4. 83 4. 50 4. 50 Temp 56, 10 min 4. 00 4. 33 4. 33 p H 3. 0, 4, 18 h 0 0 0 56, 30 min 1. 00 0.

4 3 : 85 CPE, Reed2Muench TCID , (56 ), (p H 5. 0), (p H 8. 0), RNA 214 PRRSV 3, Marc2145 PRRSV J 1 J 2 J 3 100, 3 PRRSV, Treatment methods 1 Table 1 Infectivity of isolated virus following treatment with various methods TCID 50 / ml (log10) J1 J2 J3 Treatment methods TCID 50 / ml (log10) J1 J2 J3 ( = 5 %) 4,24 h , 1 h Untreated Temp 56, 10 min p H 3. 0, 4, 18 h , 30 min , 1 h ( = 012 %) 2 h Formalin 12 h BUDR(60 g/ ml) h Untreated Untreated Marc Table 2 Results of virus neutralization test TCID, 50 / ml Isolates (5 log10) Neutralization index, nm ( 2) J , nm J J ( 3) Control group Marc2145 PRRSV 3 PRRSV J1 Fig. 2 Electron micrograph of PRRSV strain J1 Fig. 3 Electron micrograph of PRRSV strain J1 in Marc2145 cells negative stain with 2 % PTA

5 PRRSV, 3,, PRRSV,, PRRSV,PRRSV,, 3,, [9 ] PRRSV, [7,8, 3, Yoon ],56 1 h, p H h = 012 % 37 1 h, ;56 30 min, p H (37 1 h) 90 % Marc h, nm,, nm ( PAM) Marc2145 CL2601 PRRSV, SPF,,CL2601,,Marc Marc , 6, ( TCID 50 ) 10 5 / ml,, 1 Wenswoort G. Porcine reproductory and respiratory syndrome. Proc, 13 th Int Pig Vet Soc Congr, Collins J E, Benfield D A, Chrisitanson W T, et al. Isolation of swine infectility and respiratory syndrome virus (isolate A TCCVR22322) in North America and experimental reproduction of the disease in gnotobiotic pigs. J Vet Diagn Invest, 1992,4 : Hopper S A, White M E C, Twiddy N. An outbreak of boue2eared pig disease (porcine reproductive and res2 piratory syndrome) in four pig herds in Great Britain. Vet Rec, 1992,15 : Kuwahara H, Nunoya T, Tajima M, et al. An outbreak of porcine reproductive and respiratory syndrome in Japan. J Vet Med Sci, 1994, 56(5) : Murakami Y, Kato A, Tsuds T, et al. Isolation and serological characterization of porcine reproductive and respiratory syndrome ( PRRS) viruses from pigs with reproductive and respiratory disorders in Japan. J Vet Med Sci, 1994,56(5) : Shimizu M, Yamada S, Murakami Y, et al. Isolation of porcine reproductive and respiratory syndrome ( PRRS) virus from Heko2Heko disease of pigs. J Vet Med Sei, 1994,56(2) : Yoon I J, Jco H S, Christianson W T. et al. Isolation of a cytopathic virus from weak pigs on farms with a history of swine infertility and respiratory syndrome. J Vet Diagn Invest, 1992,4 : ,,, 1 1 1,1993,4 : ,,, 1 PRRS ( PRRSV) 1,1996,2 : Kim K S, Kwang J, Yoon J, et al. Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogereous subpopulation of MA104 cell line. Archiv Virol, 1993,133(3 4) : ( )

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