Eradication of PRRS from Hungarian Pig Herds between 2014 and 2022.

Porcine reproductive and respiratory syndrome (PRRS) is a widespread infectious disease that is currently a major cause of economic losses in pig production. In Hungary, a National PRRS Eradication Program has been introduced to attain a more efficient, economic, and competitive international market position. The program has been also approved by the EU, but the resulting legal obligations have imposed a burden on Hungarian producers to comply with EU competition rules. The implementation of the program has been carried out by the veterinary authorities with the consent of, continuous support from and monitoring conducted by organisations within the pig sector as well as a scientific committee. The PRRS eradication program in Hungary was based on a regional territorial principle and was compulsory for all pig holdings within the regions. In Hungary, large fattening farms operate as all-in/all-out or continuous flow systems. Large-scale breeding herds are predominantly farrow-to-finish types. Although its significance has decreased in recent decades, 20% of the Hungarian pig population is still kept on small (backyard) farms (<100 animals). All PRRSV-infected large-scale farms had to develop a unit-adapted eradication plan, including external and internal biosecurity measures, vaccinations, etc. It was crucial to render each fattening unit free of the disease, as fattening units play a significant role in spreading the virus within the country. The eradication efforts mainly implemented were depopulation-repopulation methods, but on some farms a testing and removal method has been used. As the eradication progressed over the years, the introduction of infected fattening pigs was restricted. Thanks to these measures, Hungarian large-scale fattening farms became PRRSV-free by the end of 2018. The PRRSV-free status of small-scale herds was achieved by the end of 2015 and was maintained between 2016 and 2021. By 31 December 2021, all breeding pigs in large-scale farms in Hungary were free of wild-type PRRS virus. By 31 March 2022, the total pig population of the country, including all backyard farms and fattening units, achieved PRRSV-free status. The future goal is to ensure and maintain the PRRSV-free status of Hungary via strict import regulations of live animals combined with the continuous and thorough screening of incoming and resident herds for the presence of the virus.

Quantitative Analysis of Inflammatory Uterine Lesions of Pregnant Gilts with Digital Image Analysis Following Experimental PRRSV-1 Infection.

Reproductive disorders caused by porcine reproductive and respiratory syndrome virus-1 are not yet fully characterized. We report QuPath-based digital image analysis to count inflammatory cells in 141 routinely, and 35 CD163 immunohistochemically stained endometrial slides of vaccinated or unvaccinated pregnant gilts inoculated with a high or low virulent PRRSV-1 strain. To illustrate the superior statistical feasibility of the numerical data determined by digital cell counting, we defined the association between the number of these cells and endometrial, placental, and fetal features. There was strong concordance between the two manual scorers. Distributions of total cell counts and endometrial and placental qPCR results differed significantly between examiner1's endometritis grades. Total counts' distribution differed significantly between groups, except for the two unvaccinated. Higher vasculitis scores were associated with higher endometritis scores, and higher total cell counts were expected with high vasculitis/endometritis scores. Cell number thresholds of endometritis grades were determined. A significant correlation between fetal weights and total counts was shown in unvaccinated groups, and a significant positive correlation was found between these counts and endometrial qPCR results. We revealed significant negative correlations between CD163+ counts and qPCR results of the unvaccinated group infected with the highly virulent strain. Digital image analysis was efficiently applied to assess endometrial inflammation objectively.

In Situ Hybridization of PRRSV-1 Combined with Digital Image Analysis in Lung Tissues of Pigs Challenged with PRRSV-1.

Betaarterivirus suid 1 and 2 are the causative agents of porcine reproductive and respiratory syndrome (PRRS), which is one of the most significant diseases of the swine industry, causing significant economic losses in the main pig producing countries. Here, we report the development of a novel, RNA-based in situ hybridization technique (RNAscope) to detect PRRS virus (PRRSV) RNA in lung tissues of experimentally infected animals. The technique was applied to lung tissues of 20 piglets, which had been inoculated with a wild-type, highly pathogenic PRRSV-1 strain. To determine the RNAscope's applicability as a semi-quantitative method, we analysed the association between the proportion of the virus-infected cells measured with an image analysis software (QuPath) and the outcome of the real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) tests performed in parallel. The results of the quantitative approach of these two molecular biological methods show significant association (pseudo R2 = 0.3894, p = 0.004). This is the first time RNAscope assay has been implemented for the detection of PRRSV-1 in experimental animals.