Display of porcine epidemic diarrhea virus spike protein B-cell linear epitope on Lactobacillus mucosae G01 S-layer surface induce a robust immunogenicity in mice.

The Porcine epidemic diarrhea virus (PEDV) presents a substantial risk to the domestic pig industry, resulting in extensive and fatal viral diarrhea among piglets. Recognizing the mucosal stimulation triggered by PEDV and harnessing the regulatory impact of lactobacilli on intestinal function, we have developed a lactobacillus-based vaccine that is carefully designed to elicit a strong mucosal immune response. Through bioinformatics analysis, we examined PEDV S proteins to identify B-cell linear epitopes that meet the criteria of being non-toxic, soluble, antigenic, and capable of neutralizing the virus. In this study, a genetically modified strain of Lactobacillus mucosae G01 (L.mucosae G01) was created by utilizing the S layer protein (SLP) as a scaffold for surface presentation. Chimeric immunodominant epitopes with neutralizing activity were incorporated at various sites on SLP. The successful expression of SLP chimeric immunodominant epitope 1 on the surface of L.mucosae G01 was confirmed through indirect immunofluorescence and transmission electron microscopy, revealing the formation of a transparent membrane. The findings demonstrate that the oral administration of L.mucosae G01, which expresses the SLP chimeric immunodominant gene epitope1, induces the production of secreted IgA in the intestine and feces of mice. Additionally, there is an elevation in IgG levels in the serum. Moreover, the levels of cytokines IL-2, IL-4, IFN-γ, and IL-17 are significantly increased compared to the negative control group. These results suggest that L. mucosae G01 has the ability to deliver exogenous antigens and elicit a specific mucosal immune response against PEDV. This investigation presents new possibilities for immunoprophylaxis against PEDV-induced diarrhea.

Structural and nonstructural proteins of Senecavirus A: Recent research advances, and lessons learned from those of other picornaviruses.

Senecavirus A (SVA) is an emerging virus, causing vesicular disease in swine. SVA is a single-stranded, positive-sense RNA virus, which is the only member of the genus Senecavirus in the family Picornaviridae. SVA genome encodes 12 proteins: L, VP4, VP2, VP3, VP1, 2A, 2B, 2C, 3A, 3B, 3C and 3D. The VP1 to VP4 are structural proteins, and the others are nonstructural proteins. The replication of SVA in host cells is a complex process coordinated by an elaborate interplay between the structural and nonstructural proteins. Structural proteins are primarily involved in the invasion and assembly of virions. Nonstructural proteins modulate viral RNA translation and replication, and also take part in antagonizing the antiviral host response and in disrupting some cellular processes to allow virus replication. Here, we systematically reviewed the molecular functions of SVA structural and nonstructural proteins by reference to literatures of SVA itself and other picornaviruses.

Lactate is useful for the efficient replication of porcine epidemic diarrhea virus in cell culture.

Porcine epidemic diarrhea virus (PEDV) is a deadly pathogen infecting pig herds, and has caused significant economic losses around the world. Vaccination remains the most effective way of keeping the PEDV epidemic under control. Previous studies have shown that the host metabolism has a significant impact on viral replication. In this study, we have demonstrated that two substrates of metabolic pathway, glucose and glutamine, play a key role in PEDV replication. Interestingly, the boosting effect of these compounds toward viral replication appeared to be dose-independent. Furthermore, we found that lactate, which is a downstream metabolite, promotes PEDV replication, even when added in excess to the cell culture medium. Moreover, the role of lactate in promoting PEDV was independent of the genotype of PEDV and the multiplicity of infection (MOI). Our findings suggest that lactate is a promising candidate for use as a cell culture additive for promoting PEDV replication. It could improve the efficiency of vaccine production and provide the basis for designing novel antiviral strategies.

Butyrate limits the replication of porcine epidemic diarrhea virus in intestine epithelial cells by enhancing GPR43-mediated IFN-III production.

Porcine epidemic diarrhea virus (PEDV) is a threat to the health of newborn piglets and has a significant impact on the swine industry. Short-chain fatty acids (SCFAs) are gut microbial metabolites that regulate intestinal function through different mechanisms to enhance the intestinal barrier and immune function. In this study, we aimed to determine whether butyrate displayed a better effect than other SCFAs on limiting PEDV replication in porcine intestinal epithelial cells. Mechanistically, butyrate treatment activated the interferon (IFN) response and interferon-stimulated gene (ISG) expression. Further experiments showed that inhibition of GPR43 (free fatty acid receptor 2) in intestinal epithelial cells increased virus infection and reduced antiviral effects through IFN λ response. Our findings revealed that butyrate exerts its antiviral effects by inducing GPR43-mediated IFN production in intestinal epithelial cells.

Translation of Senecavirus A polyprotein is initiated from the IRES-proximal initiation codon.

To clarify whether Senecavirus A (SVA) has the potential of alternative translation, an extra G residue was inserted into an SVA cDNA clone, resultantly generating an "AUGAUG" motif. The second AUG is the authentic SVA initiation codon, whereas the first AUG is a putative one. Subsequently, eighteen nucleotides were inserted one by one between AUG and AUG for reconstructing cDNA clones. The test of virus recovery showed that three replication-competent SVAs, whose AUG/AUG-flanked sequences were not multiples of three nucleotides, were successfully rescued from their individual cDNA clones. The wild-type SVA possesses a UUUUU motif within the polyprotein-encoding region. Sanger sequencing showed that these three replication-competent SVAs harbored one or two extra U residues in the UUUUU motif, implying that polyprotein translation was initiated from the putative AUG, and the authentic AUG would be inactivated. This is probably attributed to the lack of ribosome scanning along an SVA genome.

Biological Characteristics of Infectious Laryngotracheitis Viruses Isolated in China.

Infectious laryngotracheitis virus (ILTV) causes severe respiratory disease in chickens and results in huge economic losses in the poultry industry worldwide. To correlate the genomic difference with the replication and pathogenicity, phenotypes of three ILTVs isolated from chickens in China from 2016 to 2018 were sequenced by high-throughput sequencing. Based on the entire genome, the isolates GD2018 and SH2017 shared 99.9% nucleotide homology, while the isolate SH2016 shared 99.7% nucleotide homology with GD2018 and SH2017, respectively. Each virus genome contained 82 ORFs encoding 77 kinds of protein, 31 of which share the same amino acid sequence in the three viruses. GD2018 and SH2017 shared 57 proteins with the same amino acid sequence, while SH2016 shared 42 and 41 proteins with the amino acid sequences of GD2018 and SH2017, respectively. SH2016 propagated efficiently in allantoic fluid and on chorioallantoic membranes (CAMs) of SPF chicken embryo eggs, while GD2018 and SH2017 proliferated well only on CAMs. GD2018 propagated most efficiently on CAMs and LMH cells among three isolates. SH2016 caused serious clinical symptoms, while GD2018 and SH2017 caused mild and moderate clinical symptoms in chickens, although the sero of the chickens infected with those three isolates were all positive for anti-ILTV antibody at 14 and 21 days after challenge. Three ILTVs with high genetic homology showed significant differences in the replication in different culture systems and the pathogenicity of chickens, providing basic materials for studying the key determinants of pathogenicity of ILTV.

Molecular epidemiology of Rickettsia sp. and Coxiella burnetii collected from Hyalomma asiaticum in Bactrian camels (Camelus bactrianus) in inner Mongolia of China.

The purpose of the present study was to determine the presence of Rickettsia sp. and Coxiella burnetii in Hyalomma asiaticum collected from Bactrian camels (Camelus bactrianus) in Inner Mongolia of China. A total of 385 H. asiaticum adults collected from Bactrian camels from Alxa in Inner Mongolia from 2017 to 2018 were examined using PCR combined with sequencing. The results indicated that 18 (4.7 %) and 5 (1.3 %) samples tested positive for Rickettsia and C. burnetii, respectively. The corresponding nucleotide identities among detected genes ranged between 99.7 % and 100 % for the sequences of 17 kD and ompB from Rickettsia and 16S rRNA and FtsX from C. burnetii. Two representative strains-Rickettsia IMA-1 strain and C. burnetii IMA-1 strain-were used for subsequent analysis. NCBI BLAST and phylogenetic analyses demonstrated that the detected strain Rickettsia IMA-1 may represent a novel species of spotted fever group (SFG) rickettsiae. The C. burnetii IMA-1 strain was grouped with Coxiella burnetii str. RSA439 (GenBank: CP040059.1). In addition, the Rickettsia sp. was successfully isolated from the ticks in Vero cells incubated at 28 °C. These findings indicate that the H. asiaticum collected off Bactrian camels in Inner Mongolia carried SFG Rickettsia species and C. burnetii and could contribute to the etiology of febrile illness in animals and humans.

[Effects of understory removal on soil labile organic carbon pool in a Cinnamomum camphora plantation].

Taking a 48-year-old Cinnamomum camphora plantation in the eastern area of our subtropics as test object, this paper studied the labile organic carbon contents and their ratios to the total organic carbon (TOC) in 0-60 cm soil layer under effects of understory removal (UR). As compared with no understory removal (CK), the soil TOC and easily-oxidized carbon (EOC) contents under UR decreased, with a decrement of 4.8% - 34.1% and 27.1% - 36.2%, respectively, and the TOC and EOC contents had a significant difference in 0-10 cm and 0-20 cm layers, respectively. The water-soluble organic carbon (WSOC) (except in 0-10 cm and 10-20 cm layers) and light fraction organic matter (LFOM) under UR increaesd, but the difference was not significant. The ratio of soil WSOC to soil TOC in UR stand was higher than that in CK stand, while the ratio of soil EOC to soil TOC showed an opposite trend. In the two stands, soil WSOC, EOC, and LFOM had significant or extremely significant correlations with soil TOC, and the correlation coefficients of soil EOC and LFOM with soil TOC were higher in UR stand than in CK, but the correlation coefficient between soil WSOC and TOC was in opposite. The soil EOC, LFOM, and TOC in the two stands were significantly or extremely significantly correlated with soil nutrients, but the soil WSOC in UR stand had no significant correlations with soil hydrolyzable N, available P, exchangeable Ca, and exchangeable Mg.