국내분리 Aujeszky's disease virus의 실험적 감염 자돈에 대한 바이러스학적 연구 : Pathogenecity, excretion, distribution and immunogenicity of virus

박정우,전무형,안수환 충남대학교부설 생명공학연구소 1992 생물공학연구지 Vol.2 No.-

To investigate the etiology, pathogenicity and virological properties of NYJ-1-87 strain of Aujeszky's disease virus (ADV) that was isolated from the diseased piglet in Korea, the virus at 10^(6.0)TCID_50/0.1㎖ was inoculated intranasally and subcutaneously into 30 to 35 days-old piglets. Results obtained through the experiments were summarized as follows. 1. Ten of the infected piglets were clinically observed for 15 days. On the 2nd day post-inoculation(pi), the signs of pyrexia, anorexia and convulsion were noted. On the 4th 7th days pi, nervous signs of incoordination and intermittent spasm were shown in the most of piglets, and one out of 5 piglets infected intranasally was died with severe nervous signs at the 7th day pi. The signs became relieved on the 8th day pi and all of remainder were completely recovered on the 13th to 14th days pi. 2. In hematological study, prominent decrease in the number of total leukocyte and lymphocyte was shown in the ADV-infected piglets on the 6th day pi. On the 8th day pi, the cell numbers were slightly increased and returned to normal level on the 10th day pi. 3. Viral excretion of the ADV-inoculated piglets was examined by swabbing of nasal and oral cavities, and rectal feces. During the periods of the 3rd to11th days pi, the virus was excreted intermittently from nasal and oral cavities, and rectal feces. The nasal excretions were shown the highest virus concentration of 10^(5.2)TCID_50/0.1㎖ at the 5th day pi. 4. Recovery of the inoculated virus from various organs of the piglets that were died or experimentally slaughtered was attempted, and the virus was isolated from the tissues of brain and tonsil by the cultured cell-inoculation method. The highest recovery rate was noted in the tonsil. By indirect immunofluorescence antibody assay using ADV-monoclonal antibody, the viral antigens were detected in tissues of spleen and liver as brain and tonsil on the 7th to 9th days pi. The virus was not isolated from blood and the tissues of lung and kidney throughout the experiments. 5. Titers of virus neutralizing antibody in the piglets experimentally infected with ADV became increased after the 6th to 9th days pi in both of intranasal and subcutaneous inoculation showing the highest titers of 64 to 128 on the 29th day pi. When the antibody levels were measured by radial immunodiffusion enzyme assay, the reactive diameter was enlarged to be positive after the 4th to 6th days pi in both of intranasal and subcutaneous inoculation showing the largest diameter of 13 to 14mm on the 29th day pi.

계태아연속계대에 의한 내열성 Newcastle disease virus 분리주의 약독화

박정우,장경수,정용성,김귀현,송희종,전무형 충남대학교 수의과대학 동물의과학연구소 1997 動物醫科學硏究誌 Vol.5 No.-

In the cases of Newcastle disease(ND) outbreaks in the pheasant farms located in Chungchung area, five heat-stable Newcastle disease viruses(NDV) have been isolated from the diseased pheasants, and the various biological properities of the isolates have been reported. In this study, we attempted attenuation of a heat-stable NDV isolate, CBPI by serial subpassages in SPF chick-embryo and examined its pathogenicity and immunogenicity. When CBPI isolate was subpassaged by the 153rd levels, the heat stability was not changed as compared with that of the parent virus. However, its virulence was reduced according to increasing level of subpassage. The chicken infected with the attenuated CBPI at age of 1, 2 and 5 weeks showed no mortality with mild respiratory signs in all age groups tested, and significantly decreasing body weight gains in the 5 week-old chicken, while parent virus showed 100% mortality in all age groups. In comparison of adverse reactions between the attenuated CBPI and B1 strain in 1 week-old chicken with NDV maternal antibody(HI antibody titer>1:8). both viruses manifested no specific clinical signs, but significantly lower body weight gain in the group of chicken inoculated with CBPI. When the attenuated CBPI or B1 strains were inoculated into the 3 week-old chicken vaccinated primarily with B1 strain, both of viruses showed no specific side effects and changes in body weight. However, the HI antibody titers in the chicken inoculated with the attenuated CBPI were slightly higher than that of the chicken inoculated with B1 vaccine.

국내에서 분리된 canine parvovirus의 구조유전자 cloning과 염기서열 분석

박종현,송재영,이중복,현방훈,안수환,전무형 충남대학교 생물공학연구소 1993 생물공학연구지 Vol.3 No.-

In this study gene encoding structural proteins of a CPV isolate was cloned and saquenced to elucidate the molecular genetical properties of the canine parvoviruses isolated from the field. Six recombinant plasmids of pEP3, p1471, p2070, pEP069, pEP338 and p14711p were constructed from the map positions 22 to 98 of RF DNA to clone the VP1 and VP2 genes of CPV-V20. Sequentialy the gene comprising 3780 nucleotides were sequenced by dideoxy chain termination method. When nucleotide sequence of gene encoding the structural proteins of CPV-V20 was compared with those of other strains, CPV-N, CPV-d and CPV-780929 published previously. DNA homologies to CPV-V20 were 99.87% with CPV-NM, 99.73% with CPV-d, 96.85% with CPV-780929 AND 98.4% with FPLV-Carl, respectively. The DNA sequence data of CPV-V20 showed seven point mutations and also deletion of 135 nucleotides from the nucleotide position 4745 to 4879 located in the 3-noncoding region of CPV-N.

닭에서 오존수의 급여가 닭와포자충 감염에 미치는 영향

박배근,染谷梓,류시윤,김현수,조성환,전무형,손화영 충남대학교 수의과대학 동물의과학연구소 2001 動物醫科學硏究誌 Vol.9 No.-

Ozone is very effective in acute and chronic viral disease as a virucidal agent and in improving the general health of the animals. The effects of ozonized water supply against Cryptosporidium baileyi was studied in 2-day-old chicken. After oral inoculation of chicken with 1×10^6 oocysts, ozonized water(0.5ppm) and tap water were supplied to experimental and control group, respectively. The prepatent period of both group was 6 days postinoculation(PI). The patent period of experimental group was shorter(6-18 days) than that of control group(6-22 days). The number of oocysts in the feces were maximum on days 11 PI, indicating 1.99×10^6 oocysts/experimental chicken and 6.00×10^6 oocysts/control chicken. The number of discharged oocysts during experimental period was higher in control group than that of experimental group. In the view of febric bursa sections on 7 days PI, the number of endogenous development of C baileyi was smaller in experimental group that in control group. This results indicate that the autoinfected sporozoite were damaged by ozone. The ozonized water supply to chickens is alleviation cryptosporidiosis.

개에서 직장탈의 수술적 치료 1례

박정희,김영실,신영지,박창식,전무형,김명철,정성목 忠南大學校 獸醫科大學 動物醫科學硏究所 2005 動物醫科學硏究誌 Vol.13 No.-

A 3-year old, male Jindo dog was referred with history of tenesmus, dyschezia and recurrent rectal prolapse. On the physical examination, the rectum was prolapsed about 10 cm, congested and necrotized. Fecal incontinence and pain were observed. Complete blood count revealed a mild leukocytosis and there were no remarkable abnormalities in serum biochemistry profiles. It was diagnosed as a rectal prolapse. Rectal resection and anastomosis was performed for necrotic tissue removal. On postoperative fecal smear, protothecosis was observed. Amphotericin B was administered. There has been no evidence of recurrence of rectal prolapse for ten months.

BCG와 levamisole 투여가 canine parvovirus subunit antigen에 대한 기니픽의 면역반응에 미치는 영향

박선경,장경수,박종현,전무형 충남대학교 수의과대학 동물의과학연구소 1995 動物醫科學硏究誌 Vol.3 No.-

To investigate the effects of priming with BCG and levamisole on the immune responses of the guinea pigs inoculated with CPV-intact antigen(CPV-IA) and recombinant CPV-subunit antigen(CPV-SA), hemagglutination inhibition(HI) test and leucocytes migration inhibition(LMI) test were applied for measurement of antibody and cell-mediated responses, respectively. 1. The minimal concentration of CPV-IA and CPV-SA to evoke HI antibody in guinea pig was 1,000 HAU and 2,000 HAU, respectively. 2. As the effects of priming with BCG and levamisole were examined by HI test, double primings showed higer HI titers than single priming and the groups primed with levamisole showed higher HI antibody titers than groups primed with BCG. All of groups primed with BCG and levamisole revealed prominantly higher HI titers than the non-primed group. 3. The optimal conditions for LMI were 64 HAU for antigens, 1×10^6 cell/ml for the concentration of leucocyte and 2 hrs for antigen-leucocyte reaction time. 4. When the cellular immune responses for the guinea pig inoculated with CPV-IA were examined by LMI test, BCG-primed groups showed significantly higher %LMI than the levamisole-primed group(P<0.05). Double primings revealed a tendency of higher %LMI than single priming, but statistically not significant. All of primed groups showed significantly higher %LMI than non-primed groups (P<0.001). 5. When the cellular immune responses for the guinea pig inoculated with CPV-SA were examined by LMI test, BCG-primed groups showed a tendency of higher %LMI than the levamisole-primed groups and double primings revealed higher %LMI than single priming in both of BCG and levamisole. All of priming groups showed significantly higher %LMI than the non-primed groups (P<0.001).

병아리에서 닭와포자충에 의한 F-낭염이 SRBC에 대한 항체형성에 미치는 영향

박배근,사또시, 가바모도,전무형,김무강,류시윤 충남대학교 수의과대학 동물의과학연구소 1996 動物醫科學硏究誌 Vol.4 No.-

병아리에 닭와포자충(Cryptosporidium baileyi)의 감염에 의한 F-낭염이 면양 적혈구(SRBS)에 대한 항체 형성에 미치는 영향을 관찰하기 위하여 2일령의 병아리에 닭와포자충의 오오시스트 1 × 10^6개를 경구 접종하였다. SRBC를 경정맥 및 F-낭의 관강에 감작하고 항체형성능과 F-낭의 조직소견을 1 주일 간격으로 10주간 관찰하여 다음과 같은 결과를 얻었다. 1. 체중의 증가율은 실험 전 기간을 통하여 전반적으로 실험군이 대조군 보다 낮았다. 2. F-낭 지수(Bursa index)는 1주령 부터 5주령 까지는 1이하로 위축되었으며, 6주령 부터 1이상으로 종창되었다. 특히, 6주령의 경우 1.94로 종창의 정도가 가장 심하였다. 3. 상피세포에서 발육하고 있는 닭와포자충의 오오시스트 및 점막하직과 여포내의 호산구의 침윤은 1주령에서 4주령 까지 관찰되었으며 1주령에 가장 많은 오오시스트가 상피에서 관찰되었다. 4. 상피세포 및 FAE의 핵융해, 비만세포의 출현, 여포의 괴사 및 상피의 증생은 5주령 이후에 관찰되었다. 상피세포의 핵융해는 6주령에 가장 심하였다. 5. HA titer는 정맥내로 항원을 감작한 군에서 7주령에 최고의 역가를 보였으며 대조군이 64±35, 실험군이 44.8±15.7이였다. F-낭의 관강내로 항원을 감작한 군에서는 8주령에 대조군이 11.2±3.9, 실험군이 2.8±1.0로 실험군의 역가가 모두 낮았다. 이상의 결과로 볼 때 닭와포자충의 감염에 의한 F-낭염(bursitis)은 SRBC에 대한 항체형성능을 저해The 함을 알 수 있다.

위축성 비염(AR) 이환돈에서 분리된 Bordetella bronchiseptica의 성상

박세종,장경수,전무형 충남대학교 수의과대학 동물의과학연구소 1994 動物醫科學硏究誌 Vol.2 No.-

During 2 years from 1992 to 1994, prevalence of atrophic rhinitis(AR) in the pig herds located in the Western Chungnam was investigated, and isolation of B bronchiseptica was attempted for the pigs manifested with the clinical signs of atrophic rhinitis. The isolates were characterized and identified in aspects of biochemical properties, antigenicity, drug sensitivity and pathogenicity. The results through the experiments are summarized as follows; 1. The prevalence of clinical cases of artrophic rhinitis was recorded by 12.7% in the group that is the sows and piglets vaccinated, 28.9% in the group that is the sows only vaccinated and 39.8% in the group of the non-vaccinated groups. In th slaughter house, 53(24.8%) of 214 pigs examined exhibit the AR lesions. 2. A total of 189 strains of B bronchiseptica were isolated from the pig herds. Isolation rates were 12.6% in the group that is the sows and piglets vaccinated, 34.1% in the group that is the sows only vaccinated and 45.7% in the group of the non-vaccinated groups. Isolation rate in the specimen from the slaughter house was 93(43.5%) of 214 pigs examined. Of the AR-non-vaccinated group, the piglets aged between 61 to 90 days revealed the highest isolation rate of 58.5%. 3. Biochemical and serological propertities of 189 isolates were very similar to those of the reference B bronchiseptica phase I type, indicating that most of isolates are B bronchiseptica phase I type. 4. In antimicrobial drug susceptibility, 87.3% of 189 isolates was susceptible to chloramphenicol, 79.9%, to amikacin, 64.6%, to cephalothin and less than 35.4% to others. 5. The whole cells and bacterial filtrates of the isolates were inoculated to guinea pigs and mice. The isolates showed the high pathogenicity and dermonecrotoxicity. 6. In SDS-PAGE analysis, the isolates presented the identical protein profiles to the reference standard strains.

무균성 뇌막염과 전략 뇌경색치매로 발현된 재발성 다발연골염

오형근,정두신,박형국,안무영 순천향의학연구소 2008 Journal of Soonchunhyang Medical Science Vol.14 No.1

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Pocine Adenovirus-3의 E1B Region의 鹽基序列 分析

朴鍾賢,宋載永,李重馥,玄芳勳,安東濬,車相昊,裵用泰,姜永源,Reddy, P S,全茂炯,安壽煥 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

돼지 아데노바이러스(PAV-3). 6618주의 EIB region이 包含되어 있는 map unit 4.0에서 9.7까지의 유전자에 대한 1,984 bp의 염기서열을 決定하였으며, 이 結果를 알려진 여러 아데노바이러스 유전자와 비교하여 다음과 같은 결과를 얻었다. 1. PAV-3의 EIB유전자는 10개의 ORF로 구성되어 있으며, 그 중 아데노바이러스의 단백질과 유사성이 있는 것은 ORF1, ORF2 및 ORF3이었다. ORF1은 Ad41의 19kd 와 BAV-2에서의 EIB ORF2에서의 아미노산의 一致率은 각각 32%와 31%이었다. 2. ORF2는 Ad2 55kd protein과 tupaia adenovirus 44kd protein가 각각 34%로 아미노산 一致率이 가장 높았으며, Ad41의 52kd protein. BAV-3의 EIB ORF3에서도 33%의 一致率을 보였다. 3. ORF1은 61-666 uncleotide (606 bp), ORF 2에서는 429-1,850 uncleotide (1,422 bp)의 부위로 각각 202, 474 a.a로 構成되었으며, 예상되는 분자량은 20 kd와 52 kd이었다. 4. ORF3는 hexon-associated pIX유전자로 추정되며 내부에 1개의 polyadenylation signal(ATAAA)이 1938-1942 uncleotide에 위치하였으며, 이 부위는 TATA box (1937-1942 uncleotide)와 중복되어 존재하였다. Porcine adenovirus type 3 (PAV-3) does not cause severe infection in pigs. Adenovirus has been suggestive of live vaccine vector carrying foreign gene. One of insertion regions is delayed early (EIB) region. However, EIB region of PAV-3 has not been molecularly characterized to date. Nucleotide sequence of EIB of PAV-3 was determined. The EIB region was composed of 1,984 bp and located between 4.0 and 9.7 map units. Three potential open reading frames(ORFs) with low level of homology to other adenoviruses and a polyadenylation signal were identified in the rightward direction of genome. The nucleotide and the predicted amino acid sequences of EIB were compared to those of human and animal adenoviruses. One of the three potential ORFs. ORF1 encoded a polypeptide homologous to bovine adenovirus type 2(BAV-2) ORF2 and human adenovirus type 41(Ad41) 19 kd protein. ORF2 encoded a polypeptide homologous to human adenovirus type 2(Ad2) 55 kd protein, bovine adenovirus type 3(BAV-3) ORF3 and porcine adenovirus type 4(PAV-4) ORF2. The predicted protein of ORF1 had homology to those of Ad41 and BAV-2 with 32 and 31% respectively, whereas the deduced protein of ORF2 had homology to those of Ad2. BAV-3 and PAV-4 with 34, 33 and 29%, respectively.