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  • Original ArticleMarch 30, 2022

    3 671 116

    Development of a real-time polymerase chain reaction assay for reliable detection of a novel porcine circovirus 4 with an endogenous internal positive control

    Hye-Ryung Kim , Jonghyun Park , Ji-Hoon Park , Jong-Min Kim , Ji-Su Baek , Da-Young Kim , Young S. Lyoo , Choi-Kyu Park

    Korean J. Vet. Serv. 2022; 45(1): 1-11 https://doi.org/10.7853/kjvs.2022.45.1.1
    Abstract

    Abstract : A novel porcine circovirus 4 (PCV4) was recently identified in Chinese and Korean pig herds. Although several conventional polymerase chain reaction (cPCR) and real-time PCR (qPCR) assays were used for PCV4 detection, more sensitive and reliable qPCR assay is needed that can simultaneously detect PCV4 and internal positive control (IPC) to avoid false-negative results. In the present study, a duplex qPCR (dqPCR) assay was developed using primers/probe sets targeting the PCV4 Cap gene and pig (glyceraldehyde-3-phosphate dehydrogenase) GAPDH gene as an IPC. The developed dqPCR assay was specifically detected PCV4 but not other PCVs and porcine pathogens, indicating that the newly designed primers/probe set is specific to the PCV4 Cap gene. Furthermore, GAPDH was stably amplified by the dqPCR in all tested viral and clinical samples containing pig cellular materials, indicating the high reliability of the dqPCR assay. The limit of detection of the assay 5 copies of the target PCV4 genes, but the sensitivity of the assay was higher than that of the previously described assays. The assay demonstrated high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 1.0%. Clinical evaluation using 102 diseased pig samples from 18 pig farms showed that PCV4 circulated in the Korean pig population. The detection rate of PCV4 obtained using the newly developed dqPCR was 26.5% (27/102), which was higher than that obtained using the previously described cPCR and TaqMan probe-based qPCR and similar to that obtained using the previously described SYBR Green-based qPCR. The dqPCR assay with IPC is highly specific, sensitive, and reliable for detecting PCV4 from clinical samples, and it will be useful for etiological diagnosis, epidemiological study, and control of the PCV4 infections.

  • Original ArticleMarch 30, 2022

    0 532 102
    Abstract

    Abstract : African swine fever (ASF) is fatal to domestic pigs and wild boars (Sus scrofa) and affects the domestic pig industry. ASF is transmitted directly through the secretions of infected domestic pigs or wild boars, an essential source of infection in disease transmission. ASFV is also very stable in the environment. Thus, the virus is detected in the surrounding environment where ASF-infected carcasses are found. In this study, ASF infection monitoring was conducted on the swab and whole blood samples from wild animals, various hematopoietic arthropod samples that could access infected wild boar carcasses or habitats to cause maintenance and spread of disease, and soil samples of wild boar habitats. ASF viral DNA detection was confirmed negative in 317 wildlife and environmental samples through a real-time polymerase chain reaction. However, ASF occurs in the wild boars and spreads throughout the Korean peninsula. Therefore, it is necessary to trace the route of ASF virus infection by a continuous vector. Additional monitoring of various samples with potential ASF infection is needed to help the epidemiologic investigation and disease prevention.

  • Original ArticleMarch 30, 2022

    1 369 94

    Simple and rapid colorimetric detection of African swine fever virus by loop-mediated isothermal amplification assay using a hydroxynaphthol blue metal indicator

    Ji-Hoon Park , Hye-Ryung Kim , Ha-Kyung Chae , Jonghyun Park , Bo-Young Jeon , Young S. Lyoo , Choi-Kyu Park

    Korean J. Vet. Serv. 2022; 45(1): 19-30 https://doi.org/10.7853/kjvs.2022.45.1.19
    Abstract

    Abstract : In this study, a simple loop-mediated isothermal amplification (LAMP) combined with visual detection method (vLAMP) assay was developed for the rapid and specific detection of African swine fever virus (ASFV), overcoming the shortcomings of previously described LAMP assays that require additional detection steps or pose a cross-contamination risk. The assay results can be directly detected by the naked eye using hydroxynaphthol blue after incubation for 40 min at 62℃. The assay specifically amplified ASFV DNA and no other viral nucleic acids. The limit of detection of the assay was <50 DNA copies/reaction, which was ten times more sensitive than conventional polymerase chain reaction (cPCR) and comparable to real-time PCR (qPCR). For clinical evaluation, the ASFV detection rate of vLAMP was higher than cPCR and comparable to OIE-recommended qPCR, showing 100% concordance, with a κ value (95% confidence interval) of 1 (1.00∼1.00). Considering the advantages of high sensitivity and specificity, no possibility for cross-contamination, and being able to be used as low-cost equipment, the developed vLAMP assay will be a valuable tool for detecting ASFV from clinical samples, even in resource-limited laboratories.

  • Original ArticleMarch 30, 2022

    0 459 91

    Molecular characterization of H3N2 influenza A virus isolated from a pig by next generation sequencing in Korea

    Yeonsu Oh , Sung-Hyun Moon , Young-Seung Ko , Eun-Jee Na , Dong-Seob Tark , Jae-Ku Oem , Won-Il Kim , Chaekwang Rim , Ho-Seong Cho

    Korean J. Vet. Serv. 2022; 45(1): 31-38 https://doi.org/10.7853/kjvs.2022.45.1.31
    Abstract

    Abstract : Swine influenza (SI) is an important respiratory disease in pigs and epidemic worldwide, which is caused by influenza A virus (IAV) belonging to the family of Orthomyxoviridae. As seen again in the 2009 swine-origin influenza A H1N1 pandemic, pigs are known to be susceptible to swine, avian, and human IAVs, and can serve as a ‘mixing vessel’ for the generation of novel IAV variants. To this end, the emergence of swine influenza viruses must be kept under close surveillance. Herein, we report the isolation and phylogenetic study of a swine IAV, A/swine/Korea/21810/2021 (sw21810, H3N2 subtype). BLASTN sequence analysis of 8 gene segments of the isolated virus revealed a high degree of nucleotide similarity (94.76 to 100%) to porcine strains circulating in Korea and the United States. Out of 8 genome segments, the HA gene was closely related to that of isolates from cluster I. Additionally, the NA gene of the isolate belonged to a Korean Swine H1N1 origin, and the PB2, PB1, NP and NS genes of the isolate were grouped into that of the Triple reassortant swine H3N2 origin virus. The PA and M genes of the isolate belonged to 2009 Pandemic H1N1 lineage. Human infection with mutants was most common through contact with infected pigs. Our results suggest the need for periodic close monitoring of this novel swine H3N2 influenza virus from a public health perspective.

  • Original ArticleMarch 30, 2022

    0 431 82

    Re-evaluation on conversion system of BactoScan and BactoCount for raw milk in South Korea

    ESeul Kim , Jin-Hwan Kim , Yeong-Seob Byun , Dasom Park , Ha-Young Kim , Soon-Seek Yoon , Jin-San Moon

    Korean J. Vet. Serv. 2022; 45(1): 39-45 https://doi.org/10.7853/kjvs.2022.45.1.39
    Abstract

    Abstract : The total bacteria count is significantly important factor for hygienic quality in raw milk. BactoScan FMTM and BactoCount IBCTM are the automated instruments for the determination of the total bacterial count in raw milk. They have been used after calibration by standard plate count (SPC) method in South Korea since 2000. It is necessary to re-evaluate for total bacterial counter according to the improvement of milk quality and the change of milk quality grade. Therefore, this study was evaluated the conversion mode of the two machines by SPC method. We collected 921 bulk-tank milk samples throughout the concentration range of 1,000∼1,000,000 CFU/mL from June 2020 to October 2021. As a result, when compared by the SPC value, there was a slight difference in total bacterial count in BactoScan below 10,000 CFU/mL and above 200,000 CFU/mL and in BactoCount above 100,000 CFU/mL, respectively. Therefore, the conversion factor for BactoScan and the conversion equation for BactoCount were newly adjusted based on SPC value, and then the correlation coefficients (R2) was 0.85 or higher. In addition, the correlation (R2) between BactoScan and BactoCount was 0.91, which means the results were high positive correlation. These results are expected to contribute to improving the accuracy of the automated instruments for determining of total bacterial count in raw milk.

  • Original ArticleMarch 30, 2022

    0 320 88

    Analysis of inflammatory markers in blood related with the occurrence of subcutaneous abscesses in goats

    Ji-yeong Ku , Jun-Hwan Park , Seo-Ho Kim , Yong-il Cho , Chan-Lan Kim , Seung-Eon Cha , Gee-Wook Shin , Jinho Park

    Korean J. Vet. Serv. 2022; 45(1): 47-54 https://doi.org/10.7853/kjvs.2022.45.1.47
    Abstract

    Abstract : Subcutaneous abscesses, which occur mainly in goats and sheep, are lymph node abscesses caused by Corynebacterium pseudotuberculosis infection, and are divided into internal, external, and mixed types depending on the type of occurrence. While diagnostic methods for subcutaneous abscesses have been continuously studied, research reports for effective treatment and management of subcutaneous abscesses are inadequate. Therefore, this study was conducted to determine the changes in biometric information related to the inflammatory markers of goats induced by subcutaneous abscesses by infection with C. pseudotuberculosis . For this, hematological tests, analysis of inflammatory indicators, and analysis of serum proteins through electrophoresis separation of goats with healthy goats and goats inoculated with C. pseudotuberculosis to induce subcutaneous abscesses were compared and analyzed by date, and the differences and characteristics were identified periodically. As a result, in goats induced with subcutaneous abscesses, anemia findings related to a rapid decrease in red blood cell (RBC), hematocrit (HCT), and hemoglobin (Hb) were observed, and a significant increase in inflammatory cells expressed in total white blood cell (WBC), neutrophil, and monocytes was observed. And the levels of acute phase protein (APP) such as fibrinogen, haptoglobin, and serum amyloid A (SAA) were observed to increase rapidly immediately after infection. In addition, in the results of electrophoretic analysis of serum proteins, it was observed that the levels of α-globulin and β-globulin were significantly increased in goats with subcutaneous abscesses. That is, when looking at these changes, it was found that the systemic inflammatory response of goats was rapidly induced immediately after infection with the C. pseudotuberculosis pathogen. Through this study, it was possible to identify changes in the biomarkers of goats with subcutaneous abscesses, which had not been reported. Furthermore, these analyzed data are thoughts to be of great help in identifying, treating, and managing the goats of subcutaneous abscesses.

  • Original ArticleMarch 30, 2022

    0 478 167

    Assessment of coagulation function by thromboelastography in dogs with mitral valve insufficiency

    Chorok Jeong , Minwoong Seo , Ocki Chang , Jinho Park , Chul Park

    Korean J. Vet. Serv. 2022; 45(1): 55-61 https://doi.org/10.7853/kjvs.2022.45.1.55
    Abstract

    Abstract : In veterinary medicine, a variety of disease are known to cause coagulation abnormalities. Identification of these coagulation abnormalities have been relied on traditional coagulation assays(platelet concentration, aPTT, PT, D-dimer, fibrinogen) which take only a small part of the coagulation pathways rather than global hemostatic capacity. Among of the hypercoagulable diseases, cardiovascular disease, such as mitral valvular disease, was not regarded as the cause of the hypercoagulability. The value of a thromboelastography (TEG) as an early predictor of coagulopathy, especially hypercoagulability, has been founded. It was associated with decreased R and K values, and increased MA and α angle. The objective of this study was to compare thromboelastography results and those of traditional coagulation tests between twenty adult dogs with mitral insufficiency (MVI group) and eleven adult healthy dogs (Healthy group). As a results, MA values in the patients with mitral insufficiency (68.8±7.8 mm) were significantly higher than the normal patients (60.4±4.8 mm) (P value<0.05). Although a little report has been reported in veterinary medicine, platelet activation seems to be related with hypercoagulability in MVI patients in human medicine. The result of this report can support this pathophysiology in veterinary medicine. In addition to traditional coagulation assay, global assessment of coagulopathy using TEG, especially ability to detect hypercoagulability, may be useful for customized treatment in MVI patients. To achieve this, further study is needed to define pathophysiology and effect of medication.

  • Original ArticleMarch 30, 2022

    0 497 99

    Detection of etiological agents of proliferative and necrotizing pneumonia in pigs in Jeju

    Jae-Hoon Kim ">, Ji-Youl Jung , Hyoung-Seok Yang , Jae-Hoon Kim ">

    Korean J. Vet. Serv. 2022; 45(1): 63-69 https://doi.org/10.7853/kjvs.2022.45.1.63
    Abstract

    Abstract : Proliferative and necrotizing pneumonia (PNP) is a form of interstitial pneumonia that occurs in post-weaning pigs. In this study, we investigated the presence of swine influenza virus (SIV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2) and Aujeszky’s disease virus (ADV) in PNP lesions in Jeju pigs. Based on the histopathologic criteria for PNP, a total of 50 cases were selected in Jeju pigs between 2008 and 2010. Coupled with histopathological examinations, the presence of ADV and SIV by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) and PRRSV and PCV2 by immunohistochemical (IHC) methods were investigated. Based on the PCR and RT-PCR methods, ADV and SIV nucleic acids were not detected in all cases. According to IHC, PRRSV was detected in 38 of the 50 cases examined (76%) and PCV2 in 25 cases (50%). PRRSV or PCV2 were detected in 19 (38%) or 6 (12%) cases, respectively. Both PRRSV and PCV2 were identified in other 19 cases (38%). Antigens of PRRSV and PCV2 were commonly observed in the cytoplasm of macrophages and clusters of necrotic cells in alveolar cavities. The results of the present study demonstrate that PRRSV is predominantly associated with PNP in Jeju pigs. Co-infection with PRRSV and PCV2 may enhance the severity of PNP lesions in affected pigs.

KJVS
Mar 30, 2024 Vol.47 No.1, pp. 1~7

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