• Title/Summary/Keyword: virus isolation

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Epidemiological Studies of Avian Reovirus Infection in Broilers in Korea (국내 육계에서의 조류 레오바이러스 감염에 대한 역학 조사)

  • Kim, J.M.;Kim, M.J.;Song, J.S.;Mo, I.P.
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.85-99
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    • 2008
  • Avian reovirus (ARV) is a causative agent of viral arthritis/tenosynovitis, and malabsorption syndrome in broiler. The characteristics of malabsorption syndrome caused by ARV are diarrhea, poor feed conversion and stunting. Therefore, ARV infection has been recognized as one of the most important disease in the poultry industry because of economical losses. However, few study of ARV infection in broiler industry has been conducted in Korea. To evaluate the presence of ARV infection in broiler farms, epidemiological survey such as serological test and virus isolation has been conducted. For the serological survey using ELISA method, we selected five broiler farms which were located at different area and had a history of growth retardation, lameness, diarrhea and poor feathering. From these farms serum samples were collected at 1 day, 14 days and market age. All these farms had no history of vaccination against ARV. In addition to serological survey, we tried to isolate ARV from birds of designated farms at market age and collected feces and tissue samples such as cecal tonsil, intestine and liver. We were identified ARV by RT-PCR and transmissible electron microscopy. The samples were inoculated into 9-day-old embryonated eggs via the chorioallantoic membrane to observe the pock formation. For the pathogenicity test of ARV isolates, we inoculated with the isolates to the right footpad of 3-week-old SPF chicks and observed clinical signs and pathological changes for 14 days after challenge. Most broilers sampled for serological survey have maternal antibodies which were widely distributed at 1 day and decreased by 14 days. However, at the market age several broiler farms showed fairly high antibody titer against ARV. This increase of antibody titer at market age means the possible infection of ARV during the grow-out period. Among total 15 samples for the isolation of ARV. 2 samples were positive by RT-PCR and finally identified as a ARV. We inoculated these isolates in the SPF birds and observed that the antibody titer was increased from 7 days after challenge. However, we did not find any clinical signs both control and challenge groups. Based on the above results, it is clear that the ARV infection has been circulated in the broiler industry and caused significant economic losses. Further study is needed to evaluate the virulence of the isolates in the digestive system of broiler and the molecular characteristics of isolates.

Studies of the egg drop laying diseases from the mass zone layer (산란계 밀집지역의 산란저하성 질병에 관한 연구)

  • Lee Jeoung-Won;Eum Sung-Shim;Park In-Gyu;Bea Joung-Jun;Joung Dong-Suk;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.28 no.2
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    • pp.121-146
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    • 2005
  • Newcastle disease (ND), infectious bronchitis (IB), low pathogenic avian Influenza (LPAI) and fowl typhoid (FT) have been known as egg drop laying diseases because of the serious layer damage from mass zone layer. In this study, such egg drop laying diseases were investigated. To access this study, we peformed to evaluate antibody titers in serum and isolated bacteria and virus from organs and feces on May, July and September in 2003. The distribution of ND from January to May, IB and LPAI from October to February of the next year, and FT from March to September were inspected by the question survey in 21 farms. ND revealed to be positive rates of 490 to 474 $(96.7\%)$ in May, 510 to 506 $(99.2\%)$ in July and 510 to 510 $(100\%)$ in September with hemagglutination inhibition (HI) test. The mean antibody titers were 10.2, 9.9 and 10.2, respectively. With regard to IB, 484 out of 490 samples $(98.7\%)$ in May, 508 of 510 $(99.6\%)$ in July and 509 of 510 $(99.8\%)$ in September showed positive results and the mean antibody titers were gradually increased with 8.2, 9.0 and 9.4, respectively. According to HI test of LPAI, the positive results were shown in 442 of 480 $(92.1\%)$, 394 of 494 $(79.8\%)$ and 402 of 483 $(83.2\%)$ in May, July and September, respectively The mean antibody titers were decreased with 4.6, 4.3 and 4.0. The distribution of LPAI also elicited the positive rates of 480 to 475 $(99.0\%)$ in May, 494 to 485$(98.2\%)$ in July, 483 to 472 $(97.7\%)$ in September as determined by ELISA and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. Compared ELISA results with HI test of LPAI the positive results were 480 to 422 $(92.1\%),\;475(99.0\%),\;494\;to\;394 (79.8\%),\;485 (98.2\%)\;and\;483\;to\;402(83.2\%),\;472(97.7\%)$. Therefore, the positive rate determined by ELISA was higher than that of HI test with 6.9, 18.4 and $14.5\%$, respectively. When performed RT-PCR for ND using organ and feces samples, the pathotypes were detected $5(15.6\%)\;in\;May,\; 2(5.3\%) in\;July,\;2(7.1\%)$ in September but there is no samples showing positive band for LPAI. In attempt to isolate Salmonella gallinarum, bacteria were obtained from 4 cases (12.5%) in May, 9 (23.6%) in July, 5 (17.8%) in September. Thus the highest rate for isolation revealed to be shown in July When evaluated the antimicrobial susceptibility to 18 isolated strains of 5. gallinarum, bacteria were sensitive to trimethoprim/sulfamethox$(61.1\%),\;kanamycin\;(55.5\%),\;ampicillin\;(55.5\%)$ and amoxacillin/clavulanic acid $(55.5\%)$, cephalothin $(50.0\%)$, but resistant to penicillin $(88.9\%)$, streptomycin $(88.9\%)$, erythromycin $(83_4\%)$ and tetracycline $(61.1\%)$. According to HI test of ND and LPAI using captured 164 wild Korean tree sparrows (Passer nontanus), the positive rates were $47.6\%\;and\;57.3\%$, and the mean HI titers were 5.32 and 4.02, respectively. 71 $(43.2\%)\;and\;58(35.3\%)$ in captured sparrows also showed more than 4 titers for HI test to ND and LPAI, respectively However, the attempt for isolation of viruses failed in all samples.

Isolation and Characterization of Defense Genes Mediated by a Pathogen-Responsive MAPK Cascade in Tobacco (담배에서 병원균에 반응하는 MAPK 신호전달체계에 의해 매개되는 방어 유전자들의 분리 및 특성화)

  • Jang, Eun-Kyoung;Kang, Eun-Young;Kim, Young-Cheol;Cho, Baik-Ho;Yang, Kwang-Yeol
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1023-1030
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    • 2008
  • NtMEK2, which is the tobacco MAPK kinase that is upstream of SIPK and WIPK, was identified using the dexamethasone (DEX)-inducible gain-of-function transgenic system. Expression of $NtNEK2^{DD}$, a constitutively active mutant of NtNEK2, leads to HR-like cell death, which indicates that the NtMEK2-SIPK/WIPK cascade controls defense responses in tobacco. However, little is known about the downstream target substrates or defense-related genes that are regulated by the NtMEK2-SIPK/ WIPK cascade. In this study, ACP-based differential display RT-PCR was used to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade in $NtNEK2^{DD}$ transgenic plants. The results identified 6 novel differentially expressed genes (DEGs). These included pathogen induced protein 2-4 (pI2-4), monoterpene synthase 2 (MTS2), seven in absentia protein (SINA), cell death marker protein 1 (CDM1), hydroxyproline-rich glycoprotein (HRGP) and unknown genes (DEG45). The induction of these genes was confirmed by RT-PCR of samples obtained from $NtNEK2^{DD}$ plants. Additionally, when compared with other isolated DEGs, the pI2-4, CDM1 and HRGP genes were significantly up-regulated in response to treatment with salicylic acid and tobacco mosaic virus. Taken together, these results suggest that three novel DEGs were regulated by the NtMEK2-SIPK/WIPK cascade involved in disease resistance in tobacco.

A Clinical Study of Aseptic Meningitis in the Northern Area of Seoul in 2002 (2002년 서울 북부지역에서 유행한 무균성 수막염의 임상적 고찰)

  • Song, Myoung Hak;Chung, Ju Young;Kwon, Ji Won;Kim, Sang Woo
    • Pediatric Infection and Vaccine
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    • v.11 no.2
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    • pp.176-182
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    • 2004
  • Purpose : The etiologic agents of aseptic meningitis remain mostly unknown due to difficulty of viral culture and identification. There was an outbreak of aseptic meningitis in northern area of Seoul from June to August, 2002. We report the clinical features, laboratory data and causative viruses on 196 children with aseptic meningitis during this period. Methods : We retrospectively studied about clinical manifestations and laboratory findings 196 patients diagnosed as aseptic meningitis at Sanggye-Paik hospital. Virus isolation and serotype identification were performed by cell culture and reverse transcription polymerase chain reaction(RT-PCR) of the cerebrospinal fluid. Results : The male to female ratio was 1.39 : 1 and the mean age was 5.8+3 years. The clinical manifestations were fever, headache and vomiting. It occurred mostly in June, July and August. The numbers of peripheral blood leukocytes were $4,800{\sim}24,360/mm^3$. On cerebrospinal fluid examinations, leukocytes were in range of 10~2,000(mean 105)/$mm^3$, protein level in range of 15~171(mean 41.4) mg/dL and glucose level from 16~97(mean 57.9) mg/dL. Viral culture of cerebrospinal fluid showed 3 cases of Echovirus 9, 1 case of 25 and 30. In stool culture, 2 cases of Echovirus 6, 2 cases of Echovirus 13 and 1 case of Echovirus 30 were isolated. Conclusion : The etiologic viruses of the aseptic meningitis in northern area of Seoul in 2002 are presumed to be Echovirus 6, 9, 13, 25, 30.

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