• Title/Summary/Keyword: enzyme-linked immunosorbent assay(ELISA)

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Serologic survey of the ruminant bacterial infectious diseases in farmed deer and wild water deer in Jeonbuk province (사육사슴 및 야생고라니에서 소 세균성 전염병에 대한 혈청학적 연구)

  • Jo, Young-Suk;Chung, Yun-Shin;So, Seung-Young;Seol, Min-Suk;Cho, Ho-Seong;Kim, Bum-Seok;Lim, Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.33 no.3
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    • pp.249-254
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    • 2010
  • Deer can be one of the susceptible animals to bovine infectious diseases, and thus, may play a role either as a reservoir or amplifier host for spreading the diseases to other species such as cattle and goat. This study was conducted to determine the serum antibodies to bacterial infectious diseases for brucellosis, tuberculosis (TB), paratuberculosis (Johne's disease) in deer. Serum samples were randomly collected from 78 deer from 31 farms at Jeonbuk province, and 7 wild water deer from Jeonbuk wild animal treatment center during 2005 to 2007, respectively. Four farm deer (5.1%) showed antibodies to tuberculosis using Antigen Rapid Bovine TB Ab Test Kit. One elk (1.3%) and one wild water deer had antibodies for paratuberculosis. Antibody against Brucellosis was not detected in tube agglutination test (TAT) and enzyme-linked immunosorbent assay (ELISA). These data suggest that caution should be applied to inspection of velvet, deer blood and meat for human consumption from deer because of zoonotic bacterial diseases in deer. In addition, farmed deer can be a transmissible host for zoonotic disease to diary or raising farm.

Induction of Systemic Resistance against Cucumber mosaic virus in Arabidopsis thaliana by Trichoderma asperellum SKT-1

  • Elsharkawy, Mohsen Mohamed;Shimizu, Masafumi;Takahashi, Hideki;Ozaki, Kouichi;Hyakumachi, Mitsuro
    • The Plant Pathology Journal
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    • v.29 no.2
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    • pp.193-200
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    • 2013
  • Trichoderma asperellum SKT-1 is a microbial pesticide that is very effective against various diseases. Our study was undertaken to evaluate T. asperellum SKT-1 for induction of resistance against yellow strain of Cucumber mosaic virus (CMV-Y) in Arabidopsis plants. Disease severity was rated at 2 weeks post inoculation (WPI). CMV titre in Arabidopsis leaves was determined by indirect enzyme-linked immunosorbent assay (ELISA) at 2 WPI. Our results demonstrated that among all Arabidopsis plants treated with barley grain inoculum (BGI) of SKT-1 NahG and npr1 plants showed no significant reduction in disease severity and CMV titre as compared with control plants. In contrast, disease severity and CMV titre were significantly reduced in all Arabidopsis plants treated with culture filtrate (CF) of SKT-1 as compared with control plants. RT-PCR results showed increased expression levels of SA-inducible genes, but not JA/ET-inducible genes, in leaves of BGI treated plants. Moreover, expression levels of SA- and JA/ET-inducible genes were increased in leaves of CF treated plants. In conclusion, BGI treatment induced systemic resistance against CMV through SA signaling cascade in Arabidopsis plants. While, treatment with CF of SKT-1 mediated the expression of a majority of the various pathogen related genes, which led to the increased defense mechanism against CMV infection.

Comparison of Probability of Detecting Bluetongue in Quarantine Testing for the Imported Cattle with Special Focus on the Sampling Scenario (수입 소의 검역검사 수준에 따른 블루텅 검출 확률 비교)

  • Pak, Son-Il
    • Journal of Veterinary Clinics
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    • v.27 no.4
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    • pp.421-426
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    • 2010
  • In view of free from bluetongue (BT) in the domestic cattle population in Korea, the key of quarantine testing for BT virus (BTV) infection is detection of cattle previously exposed to the virus. The objective of this study was to estimate the probability of detecting a cattle infected with BTV using a stochastic modeling analysis of existing quarantine testing data. Three testing scenarios were considered in this study: serological testing of all animals in all imported lots (scenario 1), serological testing of a sample of cattle from all imported lots (scenario 2), and serological testing of 50% of imported lots (scenario 3). In scenario 2 and 3, it was assumed that cattle were sampled (sample size) within each lot to detect 5% of the cattle in each lot with a 95% confidence, taking into account diagnostic sensitivity of the ELISA (enzyme-linked immunosorbent assay). The model output was the total number of BTV-infected cattle and the prevalence of BTV infection in imported cattle from the US, Australia, Canada and Japan. Compared to the scenario 1, the probability of detecting a BTV-infected cattle was estimated to be 19% and 1.6% in scenario 2 and 3, respectively. Furthermore, the analyses showed a 95% confidence that BTV prevalence was less or equal to $9.7{\times}10^{-4}$ (median = $1.5{\times}10^{-5}$), indicating that, for the scenario 2 and 3 with serological testing for a sample of cattle, the risk of introducing an exotic strain of BTV into Korea through the importation of live cattle would not be acceptable.

Production of polyclonal anti-$\beta$-adrenergic receptor antibody and it′s effects on receptor ligand binding

  • Kim, Hee-Jin;Shin, Chan-Young;Noh, Min-Su;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.86-86
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    • 1995
  • The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently by the use of specific anti-receptor antibodies. A 14-mer peptide (from Phe102 to Leu115 of ${\beta}$2-adrenergic receptor) was synthesized and this peptide was coupled to carrier protein Keyhole Limpet Hemocyanin(KLH) by glutaraldehyde method. A 0.5mg of KLH-coupled peptide was emulsified with equal volume of complete Freund's adjuvant and injected via popliteal lymph node to each of the three Newzealnd White rabbits. Booster injections were repeated at 4 weeks interval for three times with incomplete Freund's adjuvants. One week after the final injection, serum was prepared from ear artery. Nonspecific immunoglobulins were removed by passing the serum through KLH-Sepharose 6B affinity matrix and further by incubation with bovine lung aceton powder. The titer of the antibody for synthetic peptide which was determined by enzyme linked immunosorbent assay(ELISA) was about l/l,000. The antibody produced in this study revealed 67kDa protein band in the western blot of partially purified guinea pig lung ${\beta}$-adrenergic receptor preparation. The antibody inhibited ${\beta}$-adrenergic antaginist [3H] Dihydroalprenolol binding to soluble ${\beta}$-adrenergic receptor by 25% while control sera did not show any inhibitory effects, The result of this study suggests that the peptide sequence selected in this study may play some important roles in adrenergic receptor-ligand interaction.

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Fucoidan Suppresses Prostaglandin E2 Production and Akt Activation in Lipopolysaccharide-Stimulated Porcine Peripheral Blood Mononuclear Cells

  • Park, Geon-Tae;Ahn, Changhwan;Kang, Byeong-Teck;Kang, Ji-Houn;Jeung, Eui-Bae;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.34 no.3
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    • pp.172-177
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    • 2017
  • Fucoidan, a cell wall polysaccharide found in the brown seaweed, is reported to have broad-spectrum biological activities. The objectives of this study were to examine the effect of fucoidan on prostaglandin $E_2$ ($PGE_2$) and cyclooxygenase-2 (COX-2) expression in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs) and to determine whether these effects are involved in Akt activation. The levels of $PGE_2$ production in the culture supernatants from PBMCs were determined by the enzyme-linked immunosorbent assay (ELISA) kit and the levels of COX-2 mRNA were measured by real time polymerase chain reaction (RT-PCR). Akt activity was determined by Western blot analysis. Fucoidan in LPS-$na{\ddot{i}ve}$ PBMCs has no effect on $PGE_2$ production and COX-2 mRNA expression. Furthermore, fucoidan does not affect Akt activation in LPS- $na{\ddot{i}ve}$ PBMCs. However, $PGE_2$ production and COX-2 mRNA expression on PBMCs were remarkably enhanced by LPS stimulation. Akt activity was also increased by LPS. Increasing effects of $PGE_2$ production and COX-2 mRNA expression in PBMCs induced by LPS were suppressed by addition of fucoidan. In addition, fucoidan reduced an increase in Akt activity in LPS-stimulated PBMCs. These results suggested that fucoidan exerts potent anti-inflammatory properties by suppression of $PGE_2$ production, COX-2 mRNA expression and Akt activation in LPS-stimulated PBMCs.

Seroprevalence and molecular detection of Mycoplasma gallisepticum and M. synoviae infection in broiler breeder in Jeonbuk providence, Korea (전북지역 육용종계에서 Mycoplasma gallisepticum과 M. synoviae의 항체 및 유전자 양성률 조사)

  • Kwak, Kilhan;Lee, Heungiae;Yuk, Hyunsu;Lee, Jaeuk;Lee, Kwanho;Yi, Young-Joo;Lee, Sang-Myeong
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.101-105
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    • 2016
  • The present study investigated serological and molecular prevalence of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) infection in unvaccinated broiler breeder farms in Jeonbuk providence. An enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) had been used to determine antibody titers against MG and MS, and genome of these pathogens, respectively. Seventy five percent of farms were seropositive for MG and 94% of farms were seropositive for MS. In addition, the rate of antibody positive flocks against MG were 65.3% (32/49), while the rate of positive flocks against MS were 84.2% (80/95). The geometric mean antibody titers were $802.2{\pm}626$ and $27,726.7{\pm}2426$ against MG and MS, respectively. Interestingly, none of samples was positive for MG genome by PCR, while 94% (farms), 82% (flocks) and 62.6% (broiler breeder) were positive for MS genome by PCR. These findings suggest that the prevalence of MG or MS infection could be higher than expected. Thus, strict prevention program including vaccination and environmental sanitation should be implemented to avoid disease transmission from breeder to broilers as well as transmission among broilers.

Efficacy and clinical trials of Salenvac-T, bivalent killed vaccine containing Salmonella Enteritidis and Salmonella Typhimurium (Salmonella Enteritidis 및 Salmonella Typhimurium을 함유한 이가 불활화백신인 Salenvac-T의 방어효과 및 임상연구)

  • Cho, Youngjae;Kang, Zheng-Wu;Kang, Kyung-Soo;Jeong, Seunghwan;Yoon, Hee-Jun;Suh, Seungwon;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.53 no.1
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    • pp.43-48
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    • 2013
  • Commercial bivalent killed Salmonella vaccine Salenvac-T has been used in several countries in order to prevent salmonellosis with Salmonella enterica serovars Enteritidis (SE) and Typhimurium (ST) in poultry. However, this vaccine has not been used in poultry farms in South Korea. In this study, we evaluated the efficacy of Salenvac-T vaccine to protect against the challenge of virulent SE and ST, and the effect of the vaccine on egg production and mortality in layer hens. The colonization of liver, spleen and cecum with challenged SE and ST was reduced in vaccinated chickens compared with that of unvaccinated control group. The twice vaccination with Salenvac-T induced elevated antibody responses against both SE and ST detected by enzyme-linked immunosorbent assay (ELISA). The higher average hen-day production was observed in the vaccinated layer hens than in the unvaccinated layer hens without significance. The average mortality was lower in the vaccinated layer hens during the experiment period. The antibody responses to both SE and ST were persistently detected in the vaccinated layers. In summary, vaccination with Salenvac-T reduces colonization of internal organs and induces good antibody responses, thereby results in higher performance and lower egg contamination with SE and ST in layer hens.

Allergenicity Changes in Raw Shrimp (Acetes japonicus) and Saeujeot (Salted and Fermented Shrimp) in Cabbage Kimchi due to Fermentation Conditions

  • Park, Jin-Gyu;Saeki, Hiroki;Nakamura, Atsushi;Kim, Koth-Bong-Woo-Ri;Lee, Ju-Woon;Byun, Myung-Woo;Kim, Seong-Mi;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Food Science and Biotechnology
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    • v.16 no.6
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    • pp.1011-1017
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    • 2007
  • Saeujeot (salted and fermented shrimp) and kimchi are traditional Korean fermented foods. Even though shrimp have often induced severe allergic reactions in sensitized individuals, few studies have investigated the allergenicity of shrimp. The aim of this study was to observe the changes of pH and allergenicity of raw shrimp (Acetes japonicus) and saeujeot in cabbage kimchi during fermentation using competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). Fermentation was carried out at different temperatures (25, 15, and $5^{\circ}C$). The pH of cabbage kimchi added with raw shrimp or saeujeot slowly decreased at lower temperature ($5^{\circ}C$) at the end stage of the fermentation process. The binding ability of serum obtained from patients allergic to raw shrimp against shrimp tropomyosin and saeujeot in kimchi rapidly decreased during longer fermentation periods and higher temperature ($25^{\circ}C$). In conclusion, the allergenicity of both raw shrimp and saeujeot in kimchi decreased during fermentation but the decrease in allergenicity of saeujeot was greater than observed for raw shrimp.

Effects of Buja-tang Treatment on the Early Change of the Monosodium Iodoacetate-induced Osteoarthritis in Rats (부자탕이 Monosodium Iodoacetate로 유발된 골관절염의 초기변화에 미치는 영향)

  • Kim, Ji-Young;Kim, Soon-Joong;Seo, Il-Bok;Jeong, Su-Hyeon
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.2
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    • pp.87-100
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    • 2011
  • Objectives : This study was carried out to investigate the effects of Buja-tang treatment on the early change of the monosodium iodoacetate-induced osteoarthritis in rats. Methods : Arthritis was induced by injection of monosodium iodoacetate(MIA)(0.25 mg) into both knee joint cavities of rats. Arthritic rats were divided into control(n=8) and treated(n=8) group. The control group was taken distilled water and the treated group, extracts of Buja-tang by orally for 20 days. At the end of the experiment(20 days after MIA injection), gross and histopathological examinations on the articular structures of knee joints were performed. Proteoglycan(PG) content in articular cartilages was analyzed by safranine O staining method. And also, tumor necrosis factor-$\alpha$($TNF-{\alpha}$) and interleukin-$1{\beta}$($IL-1{\beta}$) contents in synovial fluid were measured by enzyme-linked immunosorbent assay(ELISA) method. Results : 1. Body weight(g) of the treated group was increased significantly compared with control group at 15 and 20 days after injection. 2. Grossly, the degree of osteoarthritis in the treated group was alleviated compared with the control group. 3. PG content in articular cartilage of the treated group was increased significantly compared with the control group. 4. Histopathologically, osteoarthritic score of the treated group was decreased significantly compared with the control group. 5. $TNF-{\alpha}$ content in synovial fluid of the treated group was decreased significantly compared with the control group. Conclusions : On the basis of these results, we suggest that Buja-tang have inhibiting effects on the progression of arthritis in MIA-induced osteoarthritis model. And it is related to inhibiting the activity of $TNF-{\alpha}$ in osteoarthritic chodrocytes and synovial membranes.

Zinc Enhances Neutrophil Extracellular Trap Formation of Porcine Peripheral Blood Polymorphonuclear Cells through Tumor Necrosis Factor-Alpha from Peripheral Blood Mononuclear Cells

  • Heo, Ju-Haeng;Kim, Hakhyun;Kang, Byeong-Teck;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.37 no.5
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    • pp.249-254
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    • 2020
  • Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.