Newcastle disease (ND), caused by virulent Newcastle disease virus (NDV), is a contagious viral disease affecting various birds and poultry worldwide. In this project, differentially expressed (DE) circRNAs, miRNAs and mRNAs were identified by high-throughput RNA sequencing (RNA-Seq) in chicken thymus at 24, 48, 72 or 96 h post LaSota NDV vaccine injection versus pre-inoculation group. The vital terms or pathways enriched by vaccine-influenced genes were tested through KEGG and GO analysis. DE genes implicated in innate immunity were preliminarily screened out through GO, InnateDB and Reactome Pathway databases. The interaction networks of DE innate immune genes were established by STRING website. Considering the high expression of gga-miR-6631-5p across all the four time points, DE circRNAs or mRNAs with the possibility to bind to gga-miR-6631-5p were screened out. Among DE genes that had the probability to interact with gga-miR-6631-5p, 7 genes were found to be related to innate immunity. Furthermore, gga-miR-6631-5p promoted LaSota NDV replication by targeting insulin induced gene 1 (INSIG1) in DF-1 chicken fibroblast cells. Taken together, our data provided the comprehensive information about molecular responses to NDV LaSota vaccine in Chinese Partridge Shank Chickens and elucidated the vital roles of gga-miR-6631-5p/INSIG1 axis in LaSota NDV replication.

Objective: The study was conducted to assess the effect of supplementation with Lactobacillus lactis (LL) on growth performance, hematological parameters, meat quality and intestinal flora in pigs from growing until slaughter. Methods: A total of 72 growing pigs (30.46 ± 3.08 kg) were randomly assigned to 3 groups (including 3 pens for each group, with 8 pigs in each pen). The three treatments comprised a basal diet (O-0) and two experimental diets supplemented for 14 weeks with 0.01% (O-100) and 0.03% (O-300) LL, respectively. Results: The final body weights of the pigs in the O-100 and O-300 groups were significantly higher (p < 0.05) than those of the O-0 group. In the grower phase, the average daily weight gain (ADG) and average daily feed intake (ADFI) of pigs fed the O-300 diet were higher (p < 0.05) than those of pigs fed the O-0 diet during the grower phase. BUN and MDA were significantly higher (p < 0.05 for all) in the O-0 group than in the O-100 and O-300 groups during the grower phase. No difference (p > 0.05) was observed in the hematological parameters among the three groups during the finisher phase. Counts of LL in the stomach were significantly higher (p < 0.05) in the O-300 group than in the O-0 group. Counts of Escherichia coli in the jejunum were significantly higher (p < 0.05) in the O-0 group than in the O-300 group. Furthermore, the hardness, cohesiveness, gumminess and resilience of longissimus dorsi muscle collected from pigs fed the O-300 diet were higher (p < 0.01; p = 0.024; p = 0.003; p = 0.014, respectively) than those of tissue collected from pigs fed the O-0 diet. Conclusion: Dietary LL supplementation increased final body weight, increased ADG in the grower phase and enhanced meat quality.

The LaSota strain of Newcastle disease virus (NDV) is a commonly used vaccine to control Newcastle disease. However, improper immunization is a common reason for vaccine failure. Hence, it is imperative to thoroughly explore innate immunity-related molecular regulatory responses to the LaSota vaccine. In this text, 140 long non-coding RNAs (lncRNAs), 8 microRNAs (miRNAs), and 1514 mRNAs were identified to be differentially expressed by RNA sequencing analysis in the thymic tissues of Chinese Partridge Shank chickens after LaSota vaccine inoculation. Moreover, 70 dysregulated genes related to innate immunity were identified based on GO, Reactome pathway, and InnateDB annotations and differential expression analysis. Additionally, dysregulated lncRNAs and innate immunity-related mRNAs that could interact with dysregulated miRNAs were identified based on bioinformatics prediction analysis via the miRanda software and differential expression analysis. Among these transcripts, expression patterns of five lncRNAs, seven miRNAs, and six mRNAs were further examined by RT-qPCR assay. Both RNA-seq and RT-qPCR outcomes showed that 10 transcripts (MSTRG.22689.1, ENSGALT00000065826, ENSGALT00000059336, ENSGALT00000060887, gga-miR-6575-5p, gga-miR-6631-5p, gga-miR-1727, paraoxonase 2 (PON2), mitogen-activated protein kinase 10, and cystic fibrosis transmembrane conductance regulator (CFTR) were highly expressed, and 4 transcripts (MSTRG.188121.10, gga-miR-6655-5p, gga-miR-6548-3p, and matrix metallopeptidase 9 (MMP9) were low expressed after NDV infection. Additionally, two potential competing endogenous RNA networks (ENSGALT00000060887/gga-miR-6575-5p/PON2 or MSTRG.188121.10/gga-miR-6631-5p/MMP9) and some co-expression axes (ENSGALT00000065826/gga-miR-6631-5p, MSTRG.188121.10/gga-miR-6575-5p, MSTRG.188121.10/CFTR, ENSGALT00000060887/MMP9) were identified based on RT-qPCR and co-expression analyses. In conclusion, we identified multiple dysregulated lncRNAs, miRNAs, and mRNAs after LaSota infection and some potential regulatory networks for these dysregulated transcripts.