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Pestiviruses, RNA viruses belonging to the Flaviviridae family, have a broad host range. Their pathogenicity varies greatly and they have caused significant economic losses in animal husbandry. In this study, a novel pestivirus, porcine abortion-associated pestivirus (PAAPeV), was isolated from pigs in China in 2023. Clinically, PAAPeV causes abortions in sows and leads to congenital tremors and death in piglets. PAAPeV replicates efficiently in ST cells. Morphologically, PAAPeV virions are spherical particles with a diameter of approximately 80 nm. Phylogenetic analysis reveals that PAAPeV is closely related to Wenzhou Pipistrellus abramus pestivirus and clusters into a distinct branch, suggesting that it represents a new species: Pestivirus chinensis . Animal experiments demonstrated that PAAPeV-infected piglets and mice exhibit significant histopathological changes. In piglets, histopathological examination revealed myocarditis, hepatitis, glial vacuolation and cerebrovascular inflammation. In mice, findings included hepatic monocyte aggregation, glomerular atrophy, pulmonary edema, inflammatory cell infiltration, and capillary dilation. Viremia has been detected in both piglets and mice, with high viral genome loads found in various organs. In vitro results revealed that PAAPeV replicates in ST cells and, to a lesser extent, in human (A549 and HepG2) and monkey (Vero) cells. Overall, PAAPeV infects both pigs and mice and has the potential to infect other mammals, indicating its ability for cross-species transmission. This poses significant risks to the pig industry and public health. Strengthening monitoring and prevention efforts for PAAPeV is crucial. Our findings greatly increase the understanding of pestivirus diversity and its broader pathogen spectrum.

Atypical Porcine Pestivirus (APPV) is an emerging pathogen in pigs. The virus causes diseases associated with neurological disorders, such as congenital tremors (CT) Type A-II in piglets, leading to economic losses in the swine industry. This study investigated an outbreak of Atypical Porcine Pestivirus (APPV) on a commercial pig farm in eastern of Thailand from November 2023 to July 2024. Our results showed APPV positive in 14.18% (39/275). APPV was highly detected in nursery pigs at 40.00% (12/30) with a statistically significant ( P  < 0.05) association between age and APPV infection. Thai-APPVs ( n  = 9) were characterized by complete genome sequencing. Phylogenetic analysis of the whole genome of APPVs showed that Thai-APPVs belonged to APPV-Clade I. Moreover, the Thai-APPVs were clustered into a distinct subclade of APPV-Clade I, namely the Thai subclade, suggesting potential geographical adaptation. In conclusion, this study is the first to report the detection and genetic characterization of APPVs in a pig farm in Thailand, enhancing the understanding of its epidemiology and genetic diversity. Our findings highlight the need for APPV surveillance, particularly in young pigs, to facilitate early interventions and control measures.

The first marine pestivirus, Phocoena pestivirus (PhoPeV), isolated from harbor porpoise, has been recently described. To further characterize this unique pestivirus, its host cell tropism and growth kinetics were determined in different cell lines. In addition, the interaction of PhoPeV with innate immunity in porcine epithelial cells and the role of selected cellular factors involved in the viral entry and RNA replication of PhoPeV were investigated in comparison to closely and distantly related pestiviruses. While Bungowannah pestivirus (BuPV), a unique porcine pestivirus closely related to PhoPeV, exhibits a broad cell tropism, PhoPeV only infects cells from pigs, cattle, sheep, and cats, as has been described for classical swine fever virus (CSFV). Viral titers correlate with the amount of intracellular PhoPeV-specific RNA detected in the tested cell lines. PhoPeV replicates most efficiently in the porcine kidney cell line SK6. Pestiviruses generally counteract the cellular innate immune response by degradation of interferon regulatory factor 3 (IRF3) mediated by the viral N-terminal protease (Npro). No degradation of IRF3 and an increased expression of the type 1 interferon-stimulated antiviral protein Mx1 was observed in porcine cells infected with PhoPeV whose genome lacks the Npro encoding region. Infection of a CD46-deficient porcine cell line suggested that CD46, which is implicated in the viral entry of several pestiviruses, is not a major factor for the viral entry of PhoPeV. Moreover, the results of this study confirmed that the cellular factor DNAJC14 plays a crucial role in viral RNA replication of non-cytopathic pestiviruses, including PhoPeV.

Atypical porcine pestivirus (APPV) was first identified in 2015 in North America by high-throughput sequencing. APPV is associated with congenital tremor A-II and is widely distributed worldwide. In this study, a total of 2630 samples of domestic pigs obtained from 14 regions of Russia from 2020 to 2024 were screened for APPV presence by qRT-PCR. APPV was detected in 12 farms located in eight regions. The overall positive rate was 8.8%. It has been established that APPV has been circulating in Russian swine herds since at least 2020. The phylogenetic analysis demonstrated that the Russian isolates are variable and assigned into three clusters. The isolates from the Krasnoyarsk Krai, Belgorod, Tomsk, and Kursk regions and the Republic of Buryatia share a high nucleotide identity (94.3–98.8%) with the Hungarian strains, while the isolates from the Moscow and Pskov regions share a nucleotide identity (89.2–94.3%) with strains from the USA. The isolate from the Republic of Mordovia has a high nucleotide identity (97.1%) with the South Korean strain. In vitro studies of the Russian isolates revealed the replication of the Belgorod 151 strain in SPEV cells. Thus, this is the first large-scale study that confirms the circulation of APPV in swine herds in Russia and describes its isolation in cell culture.

Linda virus (LindaV) was first identified in a pig farm in Styria, Austria in 2015 and associated with congenital tremor (CT) type A-II in newborn piglets. Since then, only one more LindaV affected farm was retrospectively discovered 10 km away from the initially affected farm. Here, we report the recent outbreak of a novel LindaV strain in a farrow-to-finish farm in the federal state Carinthia, Austria. No connection between this farm and the previously affected farms could be discovered. The outbreak was characterized by severe CT cases in several litters and high preweaning mortality. A herd visit two months after the onset of clinical symptoms followed by a diagnostic workup revealed the presence of several viremic six-week-old nursery pigs. These animals shed large amounts of virus via feces and saliva, implying an important epidemiological role for within- and between-herd virus transmission. The novel LindaV strain was isolated and genetically characterized. The findings underline a low prevalence of LindaV in the Austrian pig population and highlight the threat when introduced into a pig herd. Furthermore, the results urge the need to better understand the routes of persistence and transmission of this enigmatic pestivirus in the pig population.

In 2013, several Austrian piglet-producing farms recorded outbreaks of action-related repetitive myoclonia in newborn piglets (“shaking piglets”). Malnutrition was seen in numerous piglets as a complication of this tremor syndrome. Overall piglet mortality was increased and the number of weaned piglets per sow decreased by more than 10% due to this outbreak. Histological examination of the CNS of affected piglets revealed moderate hypomyelination of the white substance in cerebellum and spinal cord. We detected a recently discovered pestivirus, termed atypical porcine pestivirus (APPV) in all these cases by RT-PCR. A genomic sequence and seven partial sequences were determined and revealed a 90% identity to the US APPV sequences and 92% identity to German sequences. In confirmation with previous reports, APPV genomes were identified in different body fluids and tissues including the CNS of diseased piglets. APPV could be isolated from a “shaking piglet”, which was incapable of consuming colostrum, and passaged on different porcine cells at very low titers. To assess the antibody response a blocking ELISA was developed targeting NS3. APPV specific antibodies were identified in sows and in PCR positive piglets affected by congenital tremor (CT). APPV genomes were detected continuously in piglets that gradually recovered from CT, while the antibody titers decreased over a 12-week interval, pointing towards maternally transmitted antibodies. High viral loads were detectable by qRT-PCR in saliva and semen of infected young adults indicating a persistent infection.

Pestiviruses, particularly bovine viral diarrhea virus (BVDV), cause significant economic losses worldwide. While cattle are the primary hosts for BVDV, sheep and goats can also be affected. This nationwide survey aimed to assess the prevalence, genetic characteristics, and risk factors associated with pestiviruses in sheep and goats in Kazakhstan. A one-off cross-sectional study was conducted to estimate the prevalence of pestiviruses in sheep and goats across 58 districts in 17 oblasts of Kazakhstan. A total of 2028 animals were examined using antibody ELISA, and RT-qPCR was performed on 2056 samples. Logistic regression models were used to identify potential risk factors linked to pestiviral infection. The overall prevalence of pestiviral infection in small ruminants was estimated to be 53.7% by ELISA and 2.5% by RT-qPCR. Regression analysis revealed that age, farm type, and geographic location were risk factors for pestiviral infections in small ruminants in Kazakhstan. Partial sequence analysis of the 5′-untranslated region confirmed the presence of BVDV2. Phylogenetic analysis revealed two distinct clusters of Kazakhstani BVDV2 strains, which were significantly different from known BVDV2 genotypes. No other ruminant pestiviruses were identified. The results highlight the importance of integrating small ruminants into BVDV infection control strategies to mitigate risks to livestock.

Lateral-shaking inducing neuro-degenerative agent virus (LindaV) is a novel member of the highly diverse genus Pestivirus within the family Flaviviridae . LindaV was first detected in Austria in 2015 and was associated with congenital tremor in piglets. Since then, the virus or specific antibodies have been found in a few further pig farms in Austria. However, the actual spatial distribution and the existence of reservoir hosts is largely unknown. Since other pestiviruses of pigs such as classical swine fever virus or atypical porcine pestivirus can also infect wild boar, the question arises whether LindaV is likewise present in the wild boar population. Therefore, we investigated the presence of neutralizing antibodies against LindaV in 200 wild boar samples collected in Southern Germany, which borders Austria. To establish a serological test system, we made use of the interchangeability of the surface glycoproteins and created a chimeric pestivirus using Bungowannah virus (species Pestivirus australiaense ) as synthetic backbone. The E1 and E2 glycoproteins were replaced by the heterologous E1 and E2 of LindaV resulting in the chimera BV_E1E2_LV. Viable virus could be rescued and was subsequently applied in a neutralization test. A specific positive control serum generated against the E2 protein of LindaV gave a strong positive result, thereby confirming the functionality of the test system. All wild boar samples, however, tested negative. Hence, there is no evidence that LindaV has become highly prevalent in the wild boar population in Southern Germany.

Over the last few decades, several pestiviruses have been discovered in ruminants, pigs, and, more recently, in non-ungulate hosts. Consequently, the nomenclature and taxonomy of pestiviruses have been updated. The Tunisian sheep-like pestivirus (TSV, Pestivirus N) is an additional ovine pestivirus genetically closely related to classical swine fever virus (CSFV). In this study, during a survey of pestivirus infections in ovine farms in the Lombardy region of Northern Italy, we identified and isolated a pestivirus strain from a sheep that was found to belong to Pestivirus N species based on its genomic nucleotide identity. The sheep itself and its lamb were found to be persistently infected. We performed molecular characterization and phylogenetic analysis of three viral genomic regions (a fragment of 5′-UTR, partial Npro, and the whole E2 region). In conclusion, these results confirmed circulating TSV in Northern Italy after notification in Sicily, Italy, and France. Correlation with Italian, Tunisian, and French strains showed that detection might have resulted from the trading of live animals between countries, which supports the need for health control measures.

Since the start of the mandatory nationwide bovine viral diarrhea (BVD) eradication program in Germany in 2011, the number of persistently infected (PI) animals has decreased considerably, resulting in a continuous decrease in seroprevalence. The increasingly BVD-naive cattle population could facilitate spillover infections with non-BVDV ruminant pestiviruses. Here, we report two cases in which novel pestiviruses were isolated from cattle; in both cases, the whole genome sequence showed the highest level of identity to strain “Pestivirus reindeer-1”. Both novel viruses gave positive results in BVDV diagnostic test systems, confirming that cross-reactivity is an important issue in pestivirus diagnostics. In the first case, the pestivirus was probably transmitted from sheep kept with the affected cattle, suggesting that the co-housing of small ruminants and cattle is a risk factor. The source of infection could not be determined in the second case. The occurrence of these two cases in independent cattle holdings within a relatively short time frame suggests that it would be useful to determine the presence of pestiviruses in small ruminants or even wild ruminants to better assess risk factors, especially for BVDV-free populations.