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.
Porcine reproductive and respiratory syndrome virus (PRRSV) is a significant cause of economic loss in the swine industry, yet its control remains challenging in wean-to-harvest sites. This prospective observational study followed 95 wean-to-harvest sites across six U.S. states for one production cycle. Sites were required to be PRRSV-negative or vaccinated with a modified live virus (MLV) and free of major coronaviruses. Outbreaks were defined as RT-qPCR-positive in unvaccinated sites or detection of ORF5 sequences distinct from the MLV strain. Biosecurity data were collected through a survey, and oral fluids were tested every four weeks. PRRS outbreaks occurred in 14/42 nurseries (33.3%), 8/12 wean-to-finish (66.7%), and 35/41 finishers (82.4%), with lineage 1C.5 most frequently detected. In univariate models, higher odds of outbreaks were associated with transporting pigs of unknown status (OR 9.80, 1.73–55.37), rendering (OR 6.47, 1.62–25.84), and employee cohabitation (OR 6.15, 1.51–25.09). Protective factors included exclusive pumping equipment (OR 0.07, 0.01–0.43) and overnight downtime for multi-site workers (OR 0.15, 0.04–0.56). In multivariable models, finisher sites (OR 17.47, 2.44–125.19) and greater swine site density within one mile (OR 1.62, 1.09–2.41) significantly increased outbreak odds. These results support targeted biosecurity practices, helping farmers and the swine industry reduce PRRS outbreaks.