Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves
Bovine respiratory diseases (BRD) are associated with various predisposing factors, such as physical and physiological stress factors, and bacterial and viral pathogens. These stressors and viruses suppress immune defenses, leading to bacterial growth in the upper respiratory tract and invasion of p...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/3/669 |
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| author | Yusuke Goto Kazuhiro Fukunari Tohru Suzuki |
| author_facet | Yusuke Goto Kazuhiro Fukunari Tohru Suzuki |
| author_sort | Yusuke Goto |
| collection | DOAJ |
| description | Bovine respiratory diseases (BRD) are associated with various predisposing factors, such as physical and physiological stress factors, and bacterial and viral pathogens. These stressors and viruses suppress immune defenses, leading to bacterial growth in the upper respiratory tract and invasion of pathogens into the lower respiratory tract. Therefore, continuous monitoring of the causative pathogens would contribute to the early detection of BRD. Nasal swabs and sera from 63 clinically healthy calves were continuously collected from seven farms in Iwate prefecture from 2019 to 2021. We attempted to monitor dynamics of BRD-associated pathogens by multiplex real-time RT-PCR (RT-qPCR) using their nasal swab samples. In addition, we attempted to monitor fluctuation of antibody titers against each BRD-associated pathogen by virus neutralization test (VNT) using their sera. In contrast, nasal swabs from 89 calves infected with BRD were collected from 28 farms in Iwate prefecture from 2019 to 2021. We attempted to analyze their nasal swab samples by multiplex RT-qPCR aim to detect BRD-associated pathogens that are dominant in this region. As a result, our analyses using samples from clinically healthy calves showed that positive results by multiplex RT-qPCR were closely related to a significant increase of antibody titers by VNT in bovine coronavirus (BCoV), bovine torovirus (BToV), and bovine respiratory syncytial virus (BRSV). In addition, our data exhibited that BCoV, BToV, BRSV, bovine parainfluenza virus 3, and <i>Mycoplasma bovis</i> have been more frequently detected in calves infected with BRD compared to those detected in clinically healthy calves. Moreover, the data presented herein revealed co-infections by combination multiple viral pathogens with bacterial pathogens are closely involved in the onset of BRD. Taken together, our study demonstrates multiplex RT-qPCR which can simultaneously analyze multiple pathogens, including viruses and bacteria, and is useful for the early detection of BRD. |
| first_indexed | 2024-03-11T05:47:04Z |
| format | Article |
| id | doaj.art-5d01a9e62e324445ab3ec87e426005b3 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-11T05:47:04Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-5d01a9e62e324445ab3ec87e426005b32023-11-17T14:22:40ZengMDPI AGViruses1999-49152023-03-0115366910.3390/v15030669Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy CalvesYusuke Goto0Kazuhiro Fukunari1Tohru Suzuki2Central Iwate Prefectural Livestock Health and Hygiene Center, Takizawa 020-0605, Iwate, JapanCentral Iwate Prefectural Livestock Health and Hygiene Center, Takizawa 020-0605, Iwate, JapanDivision of Zoonosis Research and Division of Hygiene Management, Sapporo Research Station, National Institute of Animal Health, NARO, Sapporo 062-0045, Hokkaido, JapanBovine respiratory diseases (BRD) are associated with various predisposing factors, such as physical and physiological stress factors, and bacterial and viral pathogens. These stressors and viruses suppress immune defenses, leading to bacterial growth in the upper respiratory tract and invasion of pathogens into the lower respiratory tract. Therefore, continuous monitoring of the causative pathogens would contribute to the early detection of BRD. Nasal swabs and sera from 63 clinically healthy calves were continuously collected from seven farms in Iwate prefecture from 2019 to 2021. We attempted to monitor dynamics of BRD-associated pathogens by multiplex real-time RT-PCR (RT-qPCR) using their nasal swab samples. In addition, we attempted to monitor fluctuation of antibody titers against each BRD-associated pathogen by virus neutralization test (VNT) using their sera. In contrast, nasal swabs from 89 calves infected with BRD were collected from 28 farms in Iwate prefecture from 2019 to 2021. We attempted to analyze their nasal swab samples by multiplex RT-qPCR aim to detect BRD-associated pathogens that are dominant in this region. As a result, our analyses using samples from clinically healthy calves showed that positive results by multiplex RT-qPCR were closely related to a significant increase of antibody titers by VNT in bovine coronavirus (BCoV), bovine torovirus (BToV), and bovine respiratory syncytial virus (BRSV). In addition, our data exhibited that BCoV, BToV, BRSV, bovine parainfluenza virus 3, and <i>Mycoplasma bovis</i> have been more frequently detected in calves infected with BRD compared to those detected in clinically healthy calves. Moreover, the data presented herein revealed co-infections by combination multiple viral pathogens with bacterial pathogens are closely involved in the onset of BRD. Taken together, our study demonstrates multiplex RT-qPCR which can simultaneously analyze multiple pathogens, including viruses and bacteria, and is useful for the early detection of BRD.https://www.mdpi.com/1999-4915/15/3/669bovine respiratory diseasemultiplex real-time RT-PCRcalfearly detectionmonitoringvirus neutralization test |
| spellingShingle | Yusuke Goto Kazuhiro Fukunari Tohru Suzuki Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves Viruses bovine respiratory disease multiplex real-time RT-PCR calf early detection monitoring virus neutralization test |
| title | Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves |
| title_full | Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves |
| title_fullStr | Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves |
| title_full_unstemmed | Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves |
| title_short | Multiplex RT-qPCR Application in Early Detection of Bovine Respiratory Disease in Healthy Calves |
| title_sort | multiplex rt qpcr application in early detection of bovine respiratory disease in healthy calves |
| topic | bovine respiratory disease multiplex real-time RT-PCR calf early detection monitoring virus neutralization test |
| url | https://www.mdpi.com/1999-4915/15/3/669 |
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