• 대한바이러스학회지
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• 제28권3호
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• pp.233-245
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• 1998

The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcription-polymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above $2^5$ hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The nucleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.

• 한국가금학회지
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• 제27권2호
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• pp.109-114
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• 2000

The objective of these experiments was to develop an attenuated thermostable Newcastle disease virus(NDV), CBP-1 strain isolated from infected pheasants. Safety, pathogenicity and protective effects against velogenic NDV were also investigated to evaluate if the attenuated NDV, CBP-1 strain could be a candidate for a new NDV vaccine strain. CBP-1 strain was passaged up to the 173 times by nine days old embryonated eggs and chicken embryo fibroblast(CEF) cell cultures. Its sensitivitly to lipid solvents and low pH, thermostability, mean death time(MDT), intracerebral pathogenicity index(ICPI) of one day old chicks and intravenous pathogenicity index(IVPI) of four weeks old chicks were examined. Safety, boosting and protective effects were tested by chicks mortality. CBP-1 NDV strain had significant thermostability at 56$\^{C}$ for 30 minutes. by hemagglutinin activity and egg infectivity test, but was not resistant to lipid solvent. It showed possibility to use as a feed or water vaccine because of the resistance to low pH. MDT, ICPI and IVPI of CBP-1 were attenuated from 51.5, 1.96, 2.60 to 112.4, 1.12, 1.45. These results implied that the 173rd passages in embryonated egg and CEF cell cultures induced a substantial attenuation of the pathogenicity of the parent virus, changing the virulence from velogenic to intermediate between mesogenic and lentogenic. After vaccination with CBP-1 at one day old by drinking water mortality was 17.5%. However, spray vaccination with B1 at one day old, CBP-1 at two weeks ild and challenge with velogenic Kyojeongwon strain at four weeks old showed 93.5% survival rate. Mortality of chicks, vaccination with 173rd passaged CBP-1 strain at one day old, two weeks old and challenge with Kyokeongwon strain at four weeks old, was 20.0%. The results of these studies indicated that partial attenuated CBP-1 strain tended to be a low safety for ND of broiler chicks and would need to be more successive attenuation.

• 한국가금학회지
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• 제39권2호
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• pp.113-120
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• 2012

본 연구를 통하여 Vero 세포에서 강독형 NDV인 Kr005주를 55대 연속 계대하여 Vero 세포에 적응된 Kr005/V 주를 작성하였다. Kr005/V 주는 Vero 세포에서 $10^{7.8}/mL$의 높은 바이러스 증식성을 보였으며, 감염세포는 cell rounding과 같은 CPE를 형성하였다. 또한 이열성(heat labile) NDV Kr005주와 달리 NDV Kr005/V 주는 내열성(thermostable)을 보였다. 계태아에서 평균 치사 시간(MDT)를 측정한 결과, 강독형인 Kr005주(49.6시간의 MDT)와 달리 Kr005/V 주는 120시간이상의 MDT를 보여 약독형 바이러스(lentogenic virus)로 분류되었다. Kr005/V 주의 HN 단백질과 F 단백질의 염기서열을 분석한 결과, HN 단백질의 443번째 아미노산에서만 변이(T에서 S)가 관찰되었다. Kr005/V 주와 Vero 세포를 사용하여 야외 닭 혈청을 대상으로 혈청중화 항체가를 측정한 결과, Kr005주와 CEF 세포를 사용하여 측정한 혈청중화 항체가와 97%의 상관성을 나타내었다.

• 한국동물위생학회지
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• 제42권4호
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• pp.245-250
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• 2019

Newcastle disease (ND) is an infectious poultry disease that caused high mortality and reduced egg production. NDVs are regularly present in the domestic duck population. And ducks play a possible role in the maintenance and transmission of NDVs. While we were monitoring the Avian Influenza, NDVs were isolated from field samples by accident. So we analysed the biological and genetic characteristics of these viruses. Lentogenic NDVs were isolated from two farms among twenty breeder duck farms. The ages of ducks were 39 weeks old in the 'A' farm and 3~72 weeks old in the 'B' farm. And they were not inoculated with the NDVs vaccine. In the biological characteristics, the both viruses which separated from the farm 'A' and 'B' were thermostable. The amino acid sequence of a site from 112 to 119 in the fusion (F) protein was 'GKQGRLIG' which has monobasic motif in the samples of both farms. And this means the separated NDVs are lentogenic. Phylogenetic analysis was performed by entire nucleotide sequence of F protein. The virus strains from the A farm (MN095239) and the B farm (MN095240) belonged to class II genotype I. Using the analysis of whole F protein nucleic acid sequence, the MN095239 (GenBank) had homology with Ulster strain about 99.95% and the MN095239 (GenBank) had homology with KR/CK/KU_LBM255/09 strain about 99.89%. NDV surveillance is needed to investigate epidemiological relationship of domestic breeder duck isolates in Korea.

• 한국가금학회지
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• 제28권2호
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• pp.83-89
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• 2001

열 안정성을 가지고 있는 한국산 꿩에서 분리된 New-castle disease virus CBP-1주는 9일령 SPF 계태아에 접종되어 $37^{\circ}C$에서 배양하는 방법으로 173번(parent주) 누대 배양되었다. $37^{\circ}C$에서 173번 누대 배양된 NDV CBP-1 주를 10 일령 계태아에 접종한 후 저온에서 ($29^{\circ}C$) 15번 (CA-15) 30번(CA-30) 누대 배양하였다. 저온순화 주인 CA-15주와 CA-30주의 이화학적 성상검사 (열 안정성 실험, 지질 용매에 대한 감수성 실험, 산성 용매에 대한 감수성 실험)와 병원성 실험(MDT, ICPI, IVPI), 온도 감수성 실험, 안전성 실험, 부스터 효과 실험, 방어효과 등을 실험하였고, $37^{\circ}C$에서 173번 누대 배양된 parent주와 비교하였다. $29^{\circ}C$에 적응된 CA-30주는 $37^{\circ}C$와 $41^{\circ}C$에서 세포 감염력이 parent주와 비교할 때 감소하였다. CA-15주와 CA-30주를 $56^{\circ}C$에서 30분, 60분, 120분 동안 처리하였을 때 이들 저온 순화주들은 혈구응집능과 세포감염력을 상실하였다. Parent주와 CA-15, CA-30주는 ethyl ether를 10분간 처리했을 때 혈구응집능과 세포 감염력을 모두 상실하였다. 그러나 parent주와 CA-15, CA-30주는 pH 3.0-glycine HCl 완충액에 60분간 처리하였을 때 혈구응집능을 가지고 있었다. Parent주의 대뇌병원성 지수와 정맥내 병원성 지수는 각각 1.12, 1.45 이었다. 그러나 CA-30주의 대뇌 병원성 지수와 정맥내 병원성 지수는 각각 0.75, 0.00으로 감소하였다. CA-30주의 안전성은 1 일령 병아리에서 실시하였고 parent주와 B-1주와 비교였다. 이들의 안전성은 치사율을 가지고 평가하였다. parent주와 CA-30주와 B-1주의 치사율은 각각 17.5, 12.0. 0.0%이었다. CA-30주가 parent주에 비해 보다 높은 안전성을 보여 주었으나 B-1주에 비해서 아직 높은 치사율을 보여 주고 있었다.