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BackgroundPorcine deltacoronavirus (PDCoV), a novel swine enteropathogenic coronavirus, challenges the global swine industry. Currently, there are no approaches preventing swine from PDCoV infection.MethodsA new PDCoV strain named JS2211 was isolated. Next, the dimer receptor binding domain of PDCoV spike protein (RBD-dimer) was expressed using the prokaryotic expression system, and a novel nanoparticle containing RBD-dimer and ferritin (SC-Fe) was constructed using the SpyTag/SpyCatcher system. Finally, the immunoprotection of RBD-Fe nanoparticles was evaluated in mice.ResultsThe novel PDCoV strain was located in the clade of the late Chinese isolate strains and close to the United States strains. The RBD-Fe nanoparticles were successfully established. Immune responses of the homologous prime-boost regime showed that RBD-Fe nanoparticles efficiently elicited specific humoral and cellular immune responses in mice. Notably, high level PDCoV RBD-specific IgG and neutralizing antibody (NA) could be detected, and the histopathological results showed that PDCoV infection was dramatically reduced in mice immunized with RBD-Fe nanoparticles.ConclusionThis study effectively developed a candidate nanoparticle with receptor binding domain of PDCoV spike protein that offers protection against PDCoV infection in mice.

The Heilongjiang-Amur River Basin is one of the largest and most complex aquatic systems in Asia, comprising diverse wetland resources. The wetland vegetation in mid-high latitude areas has high natural value and is sensitive to climate changes. In this study, we investigated the wetland vegetation cover changes and associated responses to climate change in the Heilongjiang-Amur River Basin from 2000 to 2018 based on the growing season (May to September) climate and LAI data. Our results indicated that the wetland LAI increased at 0.014 m2·m-2/yr across Heilongjiang-Amur River Basin with the regional climate showed wetting and warming trends. On a regional scale, wetland vegetation in China and Russia had positive partial correlation with solar radiation and minimum air temperature, with precipitation showing a slight lag effect. In contrast, wetland vegetation in Mongolia had positive partial correlation with precipitation. These correlations were further investigated at different climate intervals. We found the precipitation is positively correlated with LAI in the warm regions while is negatively correlated with LAI in the wet regions, indicating an increase in precipitation is beneficial for the growth of wetland vegetation in heat sufficient areas, and when precipitation exceeds a certain threshold, it will hinder the growth of wetland vegetation. In the cold regions, we found solar radiation and minimum air temperature are positively correlated with LAI, suggesting SR and minimum air temperature instead of mean air temperature and maximum air temperature play more important roles in affecting the wetland vegetation growth in the heat limited areas. The LAI was found to be negatively correlated with maximum air temperature in the arid areas, indicating excessive temperature would inhibit the wetland vegetation growth when the water is limited. Our investigation can provide a scientific foundation for the trilateral region in wetland ecosystem protection and is beneficial for a more comprehensive understanding of the responses of wetlands in the middle and high latitudes to climate change.

African swine fever is a highly lethal contagious disease of pigs for which there is no vaccine. Its causative agent African swine fever virus (ASFV) is a highly complex enveloped DNA virus encoding more than 150 open reading frames. The antigenicity of ASFV is still unclear at present. In this study, 35 proteins of ASFV were expressed by Escherichia coli, and ELISA was developed for the detection of antibodies against these proteins. p30, p54, and p22 were presented as the major antigens of ASFV, positively reacting with all five clinical ASFV-positive pig sera, and 10 pig sera experimentally infected by ASFV. Five proteins (pB475L, pC129R, pE199L, pE184L, and pK145R) reacted well with ASFV-positive sera. The p30 induced a rapid and strong antibody immune response during ASFV infection. These results will promote the development of subunit vaccines and serum diagnostic methods against ASFV.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Classical Swine Fever Virus (CSFV) are two important pathogens, which cause serious impact on swine industry worldwide. In our previous research, rPRRSV-E2, the recombinant PRRSV expressing CSFV E2 protein, could provide sufficient protection against the lethal challenge of highly pathogenic PRRSV and CSFV, and could maintained genetically stable in vitro. Here, to evaluate the virulence reversion potential risk, rPRRSV-E2 had been continuously passaged in vivo, the stability of E2 expression and virulence of the passage viruses were analyzed. The results showed that no clinical symptoms or pathological changes could be found in the inoculated groups, and there were no significant differences of viraemia among the test groups. Sequencing and IFA analysis showed that the coding gene of exogenous CSFV E2 protein existed in the passaged viruses without any sequence mutations, deletions or insertions, and could expressed steadily. It could be concluded that the foreign CSFV E2 gene in the genome of rPRRSV-E2 could be maintained genetically stable in vivo, and rPRRSV-E2 strain had relatively low level of potential risk for virulence reversion.

The Mongolian Plateau is an arid and semi-arid region with grassland as its main vegetation. It has a fragile ecosystem and is a sensitive area for global warming. The study is based on MODIS NDVI data and growth season meteorological data from 2000 to 2018, this study examined the spatial and temporal variation characteristics of grassland vegetation on the Mongolian Plateau during the growing season using trend analysis, partial correlation analysis, and residual analysis, and it explores the dual response of NDVI changes to climate and human activities. The study’s findings demonstrated that the growing season average NDVI of grassland vegetation on the plateau gradually increased from southwest to northeast during the growing season; the growing season average NDVI demonstrated a significant overall increase of 0.023/10a (p < 0.05) from 2000 to 2018, with an increase rate of 0.030/10a in Inner Mongolia and 0.019/10a in Mongolia; the area showing a significant increase in NDVI during the growing season accounted for 91.36% of the entire study area. In Mongolian Plateau grasslands during the growing season of 2000–2018, precipitation and downward surface shortwave radiation grew significantly at rates of 34.83mm/10a and 0.57 W/m2/10a, respectively, while average air temperature decreased slightly at a rate of −0.018°C/10a. Changes in meteorological factors of grassland vegetation varied by region as well, with Inner Mongolia seeing higher rates of precipitation, lower rates of average air temperature, and lower rates of downward surface shortwave radiation than Mongolia. On the Mongolian Plateau, the NDVI of grassland vegetation in the growing season showed a significant positive correlation with precipitation (0.31) and a significant negative correlation with average air temperature (−0.09) and downward surface shortwave radiation (−0.19), indicating that increased in NDVI was driven by an increase in precipitation paired with a decrease in air temperature and a decrease in surface shortwave radiation. The overall increase in NDVI caused by human activity in the grasslands of the Mongolian Plateau was primarily positive, with around 18.37% of the region being beneficial. Climate change and human activity both affect NDVI variations in Mongolian Plateau grasslands, which are spatially heterogeneous. Moderate ecological engineering and agricultural production activities are crucial for vegetation recovery. This work is crucial to further understanding surface–atmosphere interactions in arid and semi-arid regions in the context of global climate change.

Interferons (IFNs) including type I/III IFNs are the major components of the host innate immune response against porcine epidemic diarrhea virus (PEDV) infection, and several viral proteins have been identified to antagonize type I/III IFNs productions through diverse strategies. However, the modulation of PEDV infection upon the activation of the host’s innate immune response has not been fully characterized. In this study, we observed that various IFN-stimulated genes (ISGs) were upregulated significantly in a time- and dose-dependent manner in LLC-PK1 cells infected with the PEDV G2 strain FJzz1. The transcriptions of IRF9 and STAT1 were increased markedly in the late stage of FJzz1 infection and the promotion of the phosphorylation and nuclear translocation of STAT1, implicating the activation of the JAK-STAT signaling pathway during FJzz1 infection. In addition, abundant type I/III IFNs were produced after FJzz1 infection. However, type I/III IFNs and ISGs decreased greatly in FJzz1-infected LLC-PK1 cells following the silencing of the RIG-I-like receptors (RLRs), including RIG-I and MDA5, and the Toll-like receptors (TLRs) adaptors, MyD88 and TRIF. Altogether, FJzz1 infection induces the production of type-I/III IFNs in LLC-PK1 cells, in which RLRs and TLRs signaling pathways are involved, followed by the activation of the JAK-STAT signaling cascade, triggering the production of numerous ISGs to exert antiviral effects of innate immunity.

A reliable and reproducible model in vitro for swine enteric coronaviruses infection would be intestinal models that support virus replication and can be long-term cultured and manipulated experimentally. Here, we designed a robust long-term culture system for porcine intestinal organoids from the intestinal crypt or single LGR5+ stem cell by combining previously defined insights into the growth requirements of the intestinal epithelium of humans. We showed that long-term cultured swine intestinal organoids were expanded in vitro for more than 6 months and maintained the potential to differentiate into different types of cells. These organoids were successfully infected with porcine enteric coronavirus, including porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV), and were capable of supporting virus replication and progeny release. RNA-seq analysis showed robust induction of transcripts associated with antiviral signaling in response to enteric coronavirus infection, including hundreds of interferon-stimulated genes and cytokines. Moreover, gene set enrichment analysis indicated that PEDV infection could suppress the immune response in organoids. This 3D intestinal organoid model offers a long-term, renewable resource for investigating porcine intestinal infections with various pathogens.

The recombinant bivalent live vectored vaccine rPRRSV-E2 has been proved to be a favorable genetic engineering vaccine against classical swine fever (CSF) and highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS). NADC30-like strains have recently emerged in China and caused severe disease, and it is necessary to evaluate the vaccine candidate for the currently circulating viruses. This study established a good challenge model to evaluate the candidate rPRRSV-E2 vaccine in preventing infection with a representative NADC30-like strain (ZJqz21). It was shown that the challenge control piglets displayed clinical signs typical of PRRSV, including a persistent fever, dyspnea, moderate interstitial pneumonia, lymph node congestion, and viremia. In contrast, the rPRRSV-E2 vaccination significantly alleviated the clinical signs, yielded a high level of antibodies, provided adequate protection against challenge with ZJqz21, and inhibited viral shedding and the viral load in target tissues. Our results demonstrated that the recombinant bivalent live vectored vaccine strain rPRRSV-E2 can provide efficient protection against the challenge of heterologous circulating NADC30-like strain and could be a promising vaccine candidate for the swine industry.

Porcine reproductive and respiratory syndrome virus (PRRSV), which seriously endangers the world pig industry, invades host cells through receptor-mediated endocytosis involving clathrin. CD163 is an essential receptor for PRRSV during its infection of cells. The scavenger receptor cysteine-rich 5 (SRCR5) domain of the CD163 molecule is necessary for PRRSV infection, and interacts with glycoproteins GP2a and GP4 of PRRSV, allowing the virus to infect the host cells. In this study, a monoclonal antibody (mAb) against the SRCR5-6 region of porcine CD163 was developed, and the target epitope of the mAb was determined as 497TWGTVCDSDF506, which is directly adjacent to the ligand-binding pocket (LBP) domain (487-495aa) of CD163. Further study indicated that the mAb could partially block PRRSV infection of its target cells, pulmonary alveolar macrophages. The mAb developed in the study may provide a foundation of antiviral therapy for PRRSV.

Porcine reproductive and respiratory syndrome (PRRS) is one of the most devastating diseases of the global swine industry. The causative agent porcine reproductive and respiratory syndrome virus (PRRSV) was first isolated in China in 1996 and has evolved quickly during the last two decades. To fully understand virus diversity, epidemic situation in the field, and make future predictions, a total of 365 PRRSV strains were used for evolution and genome analysis in which 353 strains were isolated from mainland China. The results showed that high diversity was found among PRRSV isolates. Total PRRSV isolates could be divided into eight subgroups. Among these subgroups strains, Original HP-PRRSV, NADC30-like, and Intermediate PRRSV were the major epidemic PRRSV strains circling in the field and would play a major role in PRRS epidemic in the future. Deletions, insertions, and recombinations have occurred frequently in the PRRSV genome. Deletions were the main driving force of viral evolution before 2006 and may also contribute further to the virus' evolution in a relatively closed or low strain diversity circumstance. The recombinant strains could be divided into three groups: the Inner group, Extensional group, and Propagating group. The evolutionary directions of the isolates in the Extensional and Propagating groups have changed, and the routes of recombination in the Propagating group were analyzed and sorted into three types. The increases in recombinant strains and high rates of recombination in recent years indicate that recombination has played a very important role in the virus' evolution. Isolates, which incorporate the advantages of their parental strains, will influence PRRSV evolution and make adverse effects on PRRS control in the future.