scholarly journals Regulatory Effect of Methylation of the Porcine AQP3 Gene Promoter Region on Its Expression Level and Porcine Epidemic Diarrhea Virus Resistance

Genes ◽  
2020 ◽  
Vol 11 (10) ◽  
pp. 1167
Author(s):  
Jia-Yun Wu ◽  
Fang Wang ◽  
Zheng-Chang Wu ◽  
Sheng-Long Wu ◽  
Wen-Bin Bao
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Promoter Region ◽  
Gene Expression Regulation ◽  
Cpg Islands ◽  
Intestinal Barrier ◽  
Pcr Amplification ◽  
Intestinal Secretion ◽  
Significant Negative Correlation ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

As an important carrier for intestinal secretion and water absorption, aquaporin 3 (AQP3) is closely related to diarrhea. In this study, we investigated the mechanisms of AQP3 gene expression regulation in porcine epidemic diarrhea virus (PEDV)-induced diarrhea confirmed by PCR amplification and sequencing. Evaluation of intestinal pathology showed that diarrhea caused by PEDV infection destroyed the intestinal barrier of piglets. qPCR analysis showed that AQP3 expression in the small intestine of PEDV-infected piglets was extremely significantly decreased. qPCR and Bisulfite sequencing PCR revealed an increase in the methylation levels of both CpG islands in the AQP3 promoter region in the jejunum of PEDV-infected piglets. The methylation of mC-20 and mC-10 sites within the two CpG islands showed a significant negative correlation with AQP3 expression. Chromatin Co-Immunoprecipitation (ChIP)-PCR showed that the Sp1 transcription factor was bound to the AQP3 promoter region containing these two CpG sites. AQP3 expression was also extremely significantly reduced in Sp1-inhibited IPEC-J2 cells, indicating that abnormal methylation at the mC-20 site of CpG1 and the mC-10 site of CpG2 reduces its expression in PEDV-infected piglet jejunum by inhibiting the binding of Sp1 to the AQP3 promoter. These findings provide a theoretical basis for further functional studies of porcine AQP3.

scholarly journals Generation of recombinant Bacillus subtilis expressing Porcine Epidemic Diarrhea Virus (PEDV) S1 protein in vegetative cell

2018 ◽  
Vol 7 (2.10) ◽  
pp. 50
Author(s):  
Paradon Jorasa ◽  
Phenjun Mekvichitsaeng ◽  
Yaowaluck Maprang Roshorm
Keyword(s):  
Bacillus Subtilis ◽  
Porcine Epidemic Diarrhea Virus ◽  
Vegetative Cell ◽  
Fusion Gene ◽  
Pcr Amplification ◽  
High Yield ◽  
Efficient Production ◽  
Fusion Genes ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV) is a mucosal (gut surface) pathogen that causes severe diarrhea in piglets; thus, a vaccine capable of inducing gut-mucosal immune response is crucial for controlling PEDV infection. Bacillus subtilis has been considered a choice for vaccine delivery to the gut mucosa. In this study, we aimed to generate recombinant B. subtilis that can produce PEDV S1 protein in vegetative cell. Two promoters, PrrnO and PgsiB-PsecA, were selected for an early and high yield expression of PEDV S1 gene in B. subtilis vegetative cell and germinating spore. Promoters, PrrnO and PgsiB-PsecA, were linked to the 5’ end of the fusion gene pgsA-PEDVS1 and the fusion genes were then inserted into plasmid pDG1662. Recombinant B. subtilis strains were generated by integrating the fusion genes into B. subtilis 168 chromosome via double crossover homologous recombination. PCR amplification and amylase activity assay confirmed integration of the fusion genes into B. subtilis chromosome at amyE locus. Expression of the pgsA-PEDVS1 in B. subtilis vegetative cells germinating from spores was then studied at 2, 4, 8 and 16 hours of culture. Tested by western blot analysis, although only cleaved products of PgsA-PEDVS1 protein were observed, expression levels of pgsA-PEDVS1 under the control of both promoters were comparable at all time points. Importantly, PgsA-PEDVS1 protein could be detected as early as 2 hours after spore inoculation in LB medium. This study suggests that both PgsiB-PsecA and PrrnO promoters can be used for efficient production of PEDV S1 in germinating spore and vegetative cell and may be applicable for expression of other heterologous genes in B. subtilis vegetative cell.  

scholarly journals Identifying outbreaks of Porcine Epidemic Diarrhea virus through animal movements and spatial neighborhoods

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Gustavo Machado ◽  
Carles Vilalta ◽  
Mariana Recamonde-Mendoza ◽  
Cesar Corzo ◽  
Montserrat Torremorell ◽  
...  
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Animal Movements ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

PSVI-4 Efficacy of Medium Chain Fatty Acids and Fatty Acid-based Feed Additives as a Mitigation Strategy Against Porcine Epidemic Diarrhea Virus and Porcine Reproductive and Respiratory Syndrome Virus

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 216-217
Author(s):  
O L Harrison ◽  
G E Nichols ◽  
J T Gebhardt ◽  
Cassandra K Jones ◽  
Jason C Woodworth ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Porcine Epidemic Diarrhea Virus ◽  
Positive Control ◽  
Feed Additives ◽  
Feed Additive ◽  
Respiratory Syndrome Virus ◽  
Post Inoculation ◽  
Diarrhea Virus ◽  
Medium Chain ◽  
Porcine Epidemic Diarrhea

Abstract Recent research has demonstrated that swine viruses can be transmitted via feed. Chemical feed additives have been suggested for the mitigation of these viruses in complete feed. Therefore, the objective of this study was to evaluate the efficacy of a commercially available formaldehyde-based feed additive, medium chain fatty acid blend (MCFA), and commercially available fatty acid-based products for mitigation of porcine epidemic diarrhea virus (PEDV) and porcine reproductive and respiratory syndrome virus (PRRSV) in a feed matrix. Treatments consisted of: 1) non-treated positive control, 2) 0.33% commercial formaldehyde-based product (Sal Curb; Kemin Industries, Inc.; Des Moines, IA), 3) 0.5% MCFA blend (1:1:1 ratio of C6:0, C8:0, and C10:0, Sigma Aldrich, St. Louis, MO), 4) 0.25%, 5) 0.5%, or 6) 1% of commercial dry mono and diglyceride-based product (Furst Strike; Furst-McNess Company, Freeport, IL), 7) 0.25%, 8) 0.5%, or 9) 1% of commercial dry mono and diglyceride-based product (Furst Protect; Furst-McNess Company, Freeport, IL), 10) 0.25%, 11) 0.5%, or 12) 1% dry mono and diglyceride-based experimental product (Furst-McNess Company, Freeport, IL) with 3 replications/treatment. Treatments were applied to complete swine feed before inoculation with 106 TCID50/g of feed with PEDV or PRRSV. Post inoculation feed was held at ambient temperature for 24 h before being analyzed via qRT-PCR. The analyzed values represent the cycle threshold. Formaldehyde and MCFA decreased (P < 0.05) the detectable RNA of PEDV and PRRSV compared to all other treatments. Furst Strike, Furst Protect, and the experimental product did not significantly impact detectability of PEDV or PRRSV RNA. In conclusion, MCFA and formaldehyde treatments are effective at reducing detection of RNA from PEDV and PRRSV in feed.

scholarly journals Differential expression and correlation analysis of miRNA–mRNA profiles in swine testicular cells infected with porcine epidemic diarrhea virus

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Xiaoqian Zhang ◽  
Chang Li ◽  
Bingzhou Zhang ◽  
Zhonghua Li ◽  
Wei Zeng ◽  
...  
Keyword(s):  
Differential Expression ◽  
Porcine Epidemic Diarrhea Virus ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Bacterial Invasion ◽  
Sequencing Data ◽  
Diarrhea Virus ◽  
Comparison Groups ◽  
Porcine Epidemic Diarrhea

AbstractThe variant virulent porcine epidemic diarrhea virus (PEDV) strain (YN15) can cause severe porcine epidemic diarrhea (PED); however, the attenuated vaccine-like PEDV strain (YN144) can induce immunity in piglets. To investigate the differences in pathogenesis and epigenetic mechanisms between the two strains, differential expression and correlation analyses of the microRNA (miRNA) and mRNA in swine testicular (ST) cells infected with YN15, YN144, and mock were performed on three comparison groups (YN15 vs Control, YN144 vs Control, and YN15 vs YN144). The mRNA and miRNA expression profiles were obtained using next-generation sequencing (NGS), and the differentially expressed (DE) (p-value < 0.05) mRNA and miRNA were obtained using DESeq R package. mRNAs targeted by DE miRNAs were predicted using the miRanda algortithm. 8039, 8631 and 3310 DE mRNAs, and 36, 36, and 22 DE miRNAs were identified in the three comparison groups, respectively. 14,140, 15,367 and 3771 DE miRNA–mRNA (targeted by DE miRNAs) interaction pairs with negatively correlated expression patterns were identified, and interaction networks were constructed using Cytoscape. Six DE miRNAs and six DE mRNAs were randomly selected to verify the sequencing data by real-time relative quantitative reverse transcription polymerase chain reaction (qRT-PCR). Based on bioinformatics analysis, we discovered the differences were mostly involved in host immune responses and viral pathogenicity, including NF-κB signaling pathway and bacterial invasion of epithelial cells, etc. This is the first comprehensive comparison of DE miRNA–mRNA pairs in YN15 and YN144 infection in vitro, which could provide novel strategies for the prevention and control of PED.

scholarly journals Sequence analysis of new variants of porcine epidemic diarrhea virus in Luzon, Philippines, in 2017

2021 ◽  
Author(s):  
Saubel Ezrael A. Salamat ◽  
Therese Marie A. Collantes ◽  
Wenchie Marie L. Lumbera ◽  
Francis A. Tablizo ◽  
Christian Thomas M. Mutia ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Porcine Epidemic Diarrhea Virus ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea
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