• Title/Summary/Keyword: porcine diarrhea

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Sequence analysis and cDNA probe hybridization of the nucleocapsid(N) protein gene of transmissible gastroenteritis virus(TGEV) and porcine epidemic diarrhea virus(PEDV) (Transmissible gastroenteritis virus(TGEV)와 porcine epidemic diarrhea virus(PEDV)의 nucleocapsid(N) 단백질 유전자에 대한 염기서열 분석과 cDNA probe hybridization)

  • Park, Ji-yong;Kim, Chul-joong;Shin, Kwang-soon;Kim, Won-yong;Kang, Shien-young;Park, Yong-ho;Han, Hae-jung;Park, Yong-ha
    • Korean Journal of Veterinary Research
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    • v.35 no.3
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    • pp.515-530
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    • 1995
  • Coronaviridae에 속하는 transmissible gastroenteritis virus(TGEV)와 porcine epidemic diarrhea virus(PEDV)를 specific하게 detection할 수 있는 방법을 개발하고자 본 연구를 수행하였다. 두 바이러스 모두 RNA 바이러스이기 때문에 reverse transcription-polymerase chain reaction(RT-PCR)으로 nucleocapsid(N) protein gene의 cDNA를 증폭시켰다. SmaI으로 처리한 pTZ19R에 ligation시킨 후 염기서열을 밝히고자 sequencing하였다. 각각의 prototype virus와 비교하여 상동성을 밝혔다. 두 바이러스에 대한 cDNA probe를 제작하여 Southern blot hybridization을 실시하였다. TGEV의 경우 백신주인 P45와 병독주인 Miller strain을 사용하였다. cDNA를 증폭시키기 위해 N1/N1R과 N2/N2R 두 가지 primer를 이용한 결과, N1/N1R primer의 경우 586bp 크기의 PCR product를 얻을 수 있었고, N2/N2R primers로 582bp의 cDNA를 증폭시킬 수 있었다. PEDV 실험을 위하여 PED 임상 증상을 나타내는 분변을 이용하여 RT-PCR을 실시하였다. P2/P2R primer로 753bp의 PCR product를 얻을 수 있었다. TGEV의 두 가지 strain의 N protein gene을 sequencing하여 prototype인 Purdue strain과 염기서열 상동성을 조사한 결과, 97%이상의 높은 homology를 나타내었다. PED-V 역시 N protein gene을 sequencing하여 CV777과 염기서열 상동성을 조사한 결과 97%이상의 homology로 PEDV임을 알 수 있었다. TGEV와 PEDV의 염기서열을 비교한 결과 29%의 낮은 homology를 관찰할 수 있었다. 두 가지 바이러스의 N protein gene에 대한 cDNA probe를 제작하여 Southern blot hybridization을 한 결과, 각 바이러스에 매우 특이적 반응을 나타내었다.

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Production and characterization of monoclonal antibody against bovine coronavirus (소 코로나바이러스에 대한 단크론항체 생산과 특성)

  • Ahn, Jae-moon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.581-588
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    • 1998
  • Eight monoclonal antibodies(MAbs) against bovine coronavirus(BCV) were produced and characterized. Three MAbs(1G9, 4H12, 5C1) specific to the S glycoprotein and two HE glycoprotein-specific MAbs(2A5, 5G4) were found to neutralize the BCV in fluorescence focus neutralization(FFN) test. Two HE-specific MAbs from the neutralizing MAbs inhibited the hemagglutinating activity of the BCV. None of the N protein-specific MAbs(1C1, 5A12, 6H1) neutralized the virus infectivity. Bovine coronavirus and mouse hepatitis virus, which belong to group II coronaviruses, were differentiated from other groups of coronaviruses(porcine transmissible gastroenteritis virus, porcine epidemic diarrhea virus, canine coronavirus) by all MAbs in fluorescence antibody test(FA), but not in FFN test.

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Identification of Salmonella spp. from porcine salmonellosis by matrix-assisted laser desorption ionization-time of flight mass spectrometry

  • Yang, Hyoung-Seok;Kim, Jae-Hoon
    • Korean Journal of Veterinary Service
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    • v.41 no.2
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    • pp.105-110
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    • 2018
  • A total of 41 Salmonella (S.) strains were isolated from pigs suffered with severe watery diarrhea and were tried to identify by both matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and polymerase chain reaction (PCR) analysis. Fibrinous exudate and ulceration in the large intestine were prevalent in gross observation, and variable degrees of enteritis were observed in the histology of large intestines. Subsequent polymerase chain reaction (PCR) analyses demonstrated that 41 strains were identified as S. Typhimurium (39 strains), though 2 stains were failed to identify. Further identification was performed using both direct smear and protein extraction method by MALDI-TOF MS analyses. In terms of extraction methods, 100% (41/41) of isolates were identified to species level of S. spp. Whereas only 43.9% (18/41) were identified to species level using the direct method. These results thus suggest that rapid and accurate diagnosis of porcine salmonellosis can be guaranteed by MALDI-TOF MS combined with protein extraction method.

Detection of bovine rotavirus antigen by enzyme linked immunosorbent assay (효소면역법에 의한 소 로타바이러스 항원 검출)

  • 안재문;유기조;이용희;이종인
    • Korean Journal of Veterinary Service
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    • v.19 no.1
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    • pp.30-38
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    • 1996
  • Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

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Clinical Examination and Control Measures in a Commercial Pig Farm Persistently Infected with Porcine Epidemic Diarrhea(PED) Virus (돼지유행성설사병 지속감염 양돈장의 임상검사 및 방제대책)

  • Park, Choi-Kyu;Lee, Chang-Hee
    • Journal of Veterinary Clinics
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    • v.26 no.5
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    • pp.463-466
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    • 2009
  • A swine farm located in the Kyungpook province (designated as farm D that have been suffering from PED for several years was selected to study the etiology and the outbreak pattern of PED by clinical and laboratory examinations. Clinical examination indicated that sows exhibited signs of mastitis resulting in an inadequate transfer of lactogenic immunity against PEDV to newborn piglets. Furthermore, serological tests revealed that all sow groups and their piglets had low levels of anti-PEDV antibody. These data suggest that improper vaccination program has been indeed performed in this farm. Remarkably, despite no symptoms of PED in weaners, the presence of PEDV was identified by RT-PCR from fecal samples of weaning piglets, indicating persistent PEDV circulation in the herd. Based on these results, the following basic control schemes were executed for the control of PEDV circulation in the farm; a) A quick removal of affected pigs and disinfection of affected sheds. b) restructuring of vaccination program and employment of consultant. c) prompt treatment of mastitis and removal of poor lactogenic sows, and d) enhancement of biosecurity of farrowing house by acquisition of additional space. We evaluated risk factors and implementation of control measures in two months and were unable to found any case related to PEDV infection. Taken together, our data indicate that the method described above is effective for the control of PED outbreak in farm persistently suffering from PEDV infection.

Preventive Effects on Porcine Epidemic Diarrhea(pED) Using by PEDV Antiserum I. Serological Results, RT-PCR for Fecal and Small Intestin, FA Test (함혈청 투여에 따른 돼지 유행성 설사병 예방효과 I. 혈청학적 결과, RT-PCR 검사, 형광항체검사)

  • Chi, Yong-Zhe;Han, Jeong-Hee;Kwon, Hyuk-Moo;Hahn, Tae-Wook;Jeong, Hyun-Kyu;Park, Bong-Kyun;
    • Korean Journal of Veterinary Pathology
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    • v.6 no.1
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    • pp.19-26
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    • 2002
  • The purpose of this study was to investigate to potective effects against porcine epidemic diarrhea virus (PEDV) infection in piglets by administration of the PEDV antiserum orally at 2 hrs, 24hrs and 36hrs after birth. six piglets administered the antiserum were experimentally infected with PEDV at five-day-old. Control group were four piglets infected with PEDV only. Serum antibody titers against PEDV were examined by serum neutralization (SN) test, dectection for PEDV or PEDV antigen from feces and small intestines was tested by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunoflurescence (IFA). The results obtained were as follows; 1. The piglets administered the PEDV antiserum showed higher antibody titers than those of control group and sustained during the experimental period. 2. The detection rate of PEDV in feces and small intestines by RT-PCR were 26.2% and 16.7% in PEDV antiserum treated group and 48.1 % and 75.0% in control group, respectively. 3. The detection rate of PEDV antigen in the small intestine by IFA were 0% in PEDV antiserum treated group and 50.0% in control group, respectively. It was concluded that oral administration of antiserum against PEDV to piglets was effective in preventing PEDV infection.

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