Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities
Porcine reproductive and respiratory syndrome virus (PRRSV) remains a major concern for swine health. Isolating PRRSV is essential for identifying infectious viruses and for vaccine formulation. This study evaluated the potential of using tongue fluid (TF) from perinatal piglet mortalities for PRRSV...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-01-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/17/1/102 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832587282163433472 |
|---|---|
| author | Onyekachukwu Henry Osemeke Isadora Machado Elisa De Conti Mariah Musskopf Mafalda Pedro Mil-Homens Samuel Stutzman Baoqing Guo Thomas Petznick Gustavo De-Sousa-E Silva Phillip Gauger Jianqiang Zhang Daniel C. L. Linhares |
| author_facet | Onyekachukwu Henry Osemeke Isadora Machado Elisa De Conti Mariah Musskopf Mafalda Pedro Mil-Homens Samuel Stutzman Baoqing Guo Thomas Petznick Gustavo De-Sousa-E Silva Phillip Gauger Jianqiang Zhang Daniel C. L. Linhares |
| author_sort | Onyekachukwu Henry Osemeke |
| collection | DOAJ |
| description | Porcine reproductive and respiratory syndrome virus (PRRSV) remains a major concern for swine health. Isolating PRRSV is essential for identifying infectious viruses and for vaccine formulation. This study evaluated the potential of using tongue fluid (TF) from perinatal piglet mortalities for PRRSV isolation. Four collection protocols were tested: extracting TF from fresh tissues using phosphate-buffered saline (PBS group), extracting TF from fresh tissues using virus transportation medium (VTM group), extracting TF from freeze-thawed tissue (freeze-thaw group), and using tissue homogenates (homogenate group). Two cell lines (ZMAC and MARC-145) and primary alveolar macrophages (PAM) were evaluated for their effect on successful PRRSV isolation. An eligible PRRSV-positive unstable breeding herd in Midwestern USA was chosen for the study. Tongues were collected in 20 batches (~30 mortalities per batch). Within each batch, each tongue tissue was cut into four quarters, with each quarter randomly assigned to one of the four collection protocols and RT-qPCR tested. Virus isolation (VI) was attempted on 10 batches. The mean RT-qPCR cycle threshold (Ct) values for the PBS, VTM, freeze-thaw, and homogenate groups were 21.9, 21.8, 22.6, and 24.8, respectively. The VI success rate was 22.6%, 12.1%, 2.8%, and 2.8% in the PBS, VTM, freeze-thaw, and homogenate groups, respectively. The probability of successful VI was 3.1% and 21.0% in the MARC-145 and ZMAC cell lines, respectively, and 4.8% in the PAM cells. TF from perinatal mortalities is an option for PRRS VI, aiding in PRRSV monitoring and control programs. |
| format | Article |
| id | doaj-art-51871f8ae65548fda2039946165e0f9c |
| institution | Kabale University |
| issn | 1999-4915 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj-art-51871f8ae65548fda2039946165e0f9c2025-01-24T13:52:35ZengMDPI AGViruses1999-49152025-01-0117110210.3390/v17010102Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine MortalitiesOnyekachukwu Henry Osemeke0Isadora Machado1Elisa De Conti2Mariah Musskopf3Mafalda Pedro Mil-Homens4Samuel Stutzman5Baoqing Guo6Thomas Petznick7Gustavo De-Sousa-E Silva8Phillip Gauger9Jianqiang Zhang10Daniel C. L. Linhares11Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USARonnfeldt Farms, Lyons, NE 68038-4574, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAVeterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-3619, USAPorcine reproductive and respiratory syndrome virus (PRRSV) remains a major concern for swine health. Isolating PRRSV is essential for identifying infectious viruses and for vaccine formulation. This study evaluated the potential of using tongue fluid (TF) from perinatal piglet mortalities for PRRSV isolation. Four collection protocols were tested: extracting TF from fresh tissues using phosphate-buffered saline (PBS group), extracting TF from fresh tissues using virus transportation medium (VTM group), extracting TF from freeze-thawed tissue (freeze-thaw group), and using tissue homogenates (homogenate group). Two cell lines (ZMAC and MARC-145) and primary alveolar macrophages (PAM) were evaluated for their effect on successful PRRSV isolation. An eligible PRRSV-positive unstable breeding herd in Midwestern USA was chosen for the study. Tongues were collected in 20 batches (~30 mortalities per batch). Within each batch, each tongue tissue was cut into four quarters, with each quarter randomly assigned to one of the four collection protocols and RT-qPCR tested. Virus isolation (VI) was attempted on 10 batches. The mean RT-qPCR cycle threshold (Ct) values for the PBS, VTM, freeze-thaw, and homogenate groups were 21.9, 21.8, 22.6, and 24.8, respectively. The VI success rate was 22.6%, 12.1%, 2.8%, and 2.8% in the PBS, VTM, freeze-thaw, and homogenate groups, respectively. The probability of successful VI was 3.1% and 21.0% in the MARC-145 and ZMAC cell lines, respectively, and 4.8% in the PAM cells. TF from perinatal mortalities is an option for PRRS VI, aiding in PRRSV monitoring and control programs.https://www.mdpi.com/1999-4915/17/1/102PRRSVpostmortem tissuestongue fluidsPCRvirus isolationsurveillance |
| spellingShingle | Onyekachukwu Henry Osemeke Isadora Machado Elisa De Conti Mariah Musskopf Mafalda Pedro Mil-Homens Samuel Stutzman Baoqing Guo Thomas Petznick Gustavo De-Sousa-E Silva Phillip Gauger Jianqiang Zhang Daniel C. L. Linhares Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities Viruses PRRSV postmortem tissues tongue fluids PCR virus isolation surveillance |
| title | Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities |
| title_full | Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities |
| title_fullStr | Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities |
| title_full_unstemmed | Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities |
| title_short | Optimizing Tongue Fluid Sampling and Testing Protocols for Enhanced PRRSV Isolation from Perinatal Swine Mortalities |
| title_sort | optimizing tongue fluid sampling and testing protocols for enhanced prrsv isolation from perinatal swine mortalities |
| topic | PRRSV postmortem tissues tongue fluids PCR virus isolation surveillance |
| url | https://www.mdpi.com/1999-4915/17/1/102 |
| work_keys_str_mv | AT onyekachukwuhenryosemeke optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT isadoramachado optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT elisadeconti optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT mariahmusskopf optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT mafaldapedromilhomens optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT samuelstutzman optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT baoqingguo optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT thomaspetznick optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT gustavodesousaesilva optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT phillipgauger optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT jianqiangzhang optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities AT danielcllinhares optimizingtonguefluidsamplingandtestingprotocolsforenhancedprrsvisolationfromperinatalswinemortalities |