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학술저널
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대한바이러스학회 JOURNAL OF BACTERIOLOGY AND VIROLOGY 大韓바이러스學會誌 제1권 제1호
발행연도
1971.6
수록면
43 - 74 (32page)

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Twenty-one viral strains have been isolated in primary cultures of calf kidney cells from fecal specimens collected from cattle, apparently healthy or showing various elinical manifestations, in various parts of Japan. Of these isolates three strains, BF-20, BFS-1 and BFK-1, were examined regarding the biological, physicochemical and immunological properties in the present study. The three strains were shown to be strains of the enterovirus group on the bases of the size of virlon, the type of viral nucleic acid, the resistance to ether, chloroform and deoxycholate and the acid stability. Thus, the strains were filtrable through Sartorius membrane filters with a pore size of 50mg. The multiplication of the strains in bovine kidney cell cultures was not inhibited with 50mg/ml of 5-iodo-2'-deoxyu- ridine in the culture medium. The strains lost no infectivity by treatment with 50% anesthetic ether at room temperature for 30 minutes, with 0.5% chloroform at room temperature for 30 minutes, or vith 0.01% sodium deoxycholate at room temperature for 2 hours. The strains were very stable at pH 3 to 9 with a slight loss of infectivity at pH 2.5 when stored at 4C for 18 hours. The strains were relatively resistant to ultraviolet irradication. They were readily inactivated by heating at 56C or higher, slowly lost infectivity at 37C or room temperature, but they could be stored without any significant loss of infectivity at 4, -20 or -80C. Worthy to note is a difference in inactivation at 50C shown between BF 20 and the other two strains. When tested with undiluted infectious culture fluids, strain BF 20 showed no significant 'decrease in titer after heating at 50C for 60 minutes, whereas strains BFS-1 and BFK-1 demonstrated considerable losses of infectivity to 10-4 or 10-4 of the original titers after the heating. This difference was also observed when the infectious culture fluids were diluted tenfold with distilled water and then heated at 50C for 60 minutes.Thus, the infectious titer decreased only about tenfold with strain BF 20 in contrast with 10 to 10-fold decrease with strains BFS-1 and BFK-1 from inactivation by incubation at 50C for 60 minutes. The slight inactivation of strain BF20 at 50C was also suppressed with 1M.MgC1,. All the strains readily multiplied and induced cytopathic effect in various cell cultures: primary cell cultures of bovine kidney and testicle, swine kidney, chick embryo, and chick kidney, and cultures of bovine embryo kidney cells subcultured many times, BHK21 cells of baby hamster kidney origin and HmLu stable line cells of hamster lung origin. Serial passages were readily accomplished in these cultures and the infectivity usually reached peak titers of 10 to 10 TCID/ml in 18 to 24 hours of incubation at 34C. Strain BF20, however, showed slower growth with lower peak titers in chick kidney and chick embryo cell cultures than in the other cultures, and strains BFS-1 and BFK-1 did so in bovine testicle cell cultures. The strains induced similar cytological changes in infected cultures, consisting of rounding, shrinkage and deterioration of the cells alnd sloughing of the glass surface. Caves developed no clinical signs and symptoms when inoculated per orally with these three strains. The viruses, however, were excreted in feces for fairly long periods of time, and neutralizing antibodies were praduced, indicating that the viruses actually infected the animals. The strains showed no pathogenicity for rabbits and guinea pigs by the intraperitoneal or intravenous route. No illness was induced in mice, 24 hours to 10 days of age, inoculated intracerbrally or intraperitoneally with these strains. Blind passages were carried out by both routes at 4-day intervals, through the 3 or 4th generation. No illness occurred in none of the inoculated mice and no virus was recovered from various tissues of those rnice. Strain BFS-1 grew in developing hens eggs when inoculated into the yolk sac, or the allantoic cavity,

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