Project description:Purpose: Searching for sRNAs in Salmonella pullorum by RNA sequencing and exploring their functions.Methods: High-throughput sequencing of RNA extracted from Salmonella pullorum under normal growth conditions to detect newly discovered sRNAs, followed by experiments to verify their functions.Results: The proportion of Clean Reads of this sequencing was >65%, and the base Q30s were all above 85%, indicating that the sequencing quality is good and can be used for subsequent analysis. The sRNAscanner software predicted that 148 new sRNAs might exist on the reference genome of Salmonella fowl dysentery, and the reads obtained from sequencing were compared to the genome, and it was found that 110 out of the 148 newly predicted sRNAs could be detected.Conclusions: sRNAs are widely found in bacteria and are involved in many physiological processes. In this study, we detected new sRNAs in Salmonella pullorum by RNA-seq, which lays the foundation for the subsequent investigation of the regulatory functions of sRNAs in bacteria.
Project description:Salmonella enterica Pullorum(S. Pullorum) is one of the most important pathogens in poultry. A better understanding of the immune response and molecular modulation resulting from infection by S. Pullorum will facilitates the control of this pathogen. In this study, we determined the relationships among identified differential expressed genes (DEGs) and pathways via deeply mining microarray data from Guangxi Huang Chicken challenged with S. Pullorum.
Project description:Salmonella enterica PullorumM-oM-<M-^HS. PullorumM-oM-<M-^I is one of the most important pathogens in poultry. A better understanding of the immune response and molecular modulation resulting from infection by S. Pullorum will facilitates the control of this pathogen. In this study, we determined the relationships among identified differential expressed genes (DEGs) and pathways via deeply mining microarray data from Guangxi Huang Chicken challenged with S. Pullorum. The chicks were then sacrificed via cervical dislocation immediately after anesthesia with 150 mg/kg sodium pentobarbital. This process was repeated at 2 hours post-infection (hpi), 4 hpi, 8 hpi, 24 hpi, 3 days post-infection (dpi), 5dpi, 7dpi, 12dpi, and 21dpi.The spleens were used to carrying out the microarray experiment. After total RNA extraction and quality control for each splenic sample were performed according to the standard protocol provided by Agilent Technologies, two random mRNA samples collected from the challenged group were equally mixed to hybridize with one chicken whole genome expression chip (Agilent.SingleColor.26441M-oM-<M-^I for each time point. In the challenged group, there were three biological replications (chips) for every time point. However, in the control group at each time point only one chip was used to hybridize with equally mixed mRNA sample containing the three control samples.
Project description:The objectives of this study were to determine the protective effects of organic acids (OA) in broilers exposed to Salmonella Pullorum challenge at early stage and to explore the potential benefits of OA by metabolomics analysis. The treatment groups included non-challenged, S. Pullorum-challenged, challenged group supplemented with virginiamycin, challenged group supplemented with OA in drinking water, challenged group supplemented with OA in feed, and challenged group supplemented with OA in combination in drinking water and feed. Results showed that early Salmonella challenge induced an acute systemic infection of broilers in the starter phase, followed by the grower phase without triggering clinical signs. OA supplementation promoted growth during the grower phase, and while OA in water contributed more, the positive effects of OA in combination were comparable to those of virginiamycin supplementation in challenged birds. Furthermore, OA could modulate the systemic metabolic perturbation caused by challenge as it alleviated stress responses mediated by steroid hormone, potentially attenuated antioxidant or immune defense, and modified intestinal microbiota metabolism. These results show a metabolic mechanism that may partly explain the potential benefits of OA in Salmonella challenged birds, and may contribute to the use of OA to control or reduce S. Pullorum infection in farm animals.
Project description:This study was conducted to evaluate the effects of dietary supplemental magnolol and honokiol in broilers infected with S. pullorum. A total of 360 one-day-old broilers were selected and randomly divided into four groups with six replicates: the negative control group (CTL), S. pullorum-infected group (SP), and the S. pullorum-infected group supplemented with 300 mg/kg honokiol (SPH) or magnolol (SPM). Chicks in the SP, SPH, and SPM groups were orally treated with a 0.5 ml (4×108 CFU/mL) S. pullorum solution at 5 days of age, while chicks in the control (CTL) group received the same amount of sterilized PBS at the same age.At 14 and 21 days of age, one chick from each replicate was randomly selected to be weighed and slaughtered by jugular exsanguination after a 12-h fasting period. The ileum samples were collected to analyze the differential expression genes.
Project description:Purpose: To elucidate the survival strategies in egg white of Salmonella Pullorum, a host-restricted pathogen with vertical transmission capability. Methods: The logarithmic-phase wild-type and Δfim mutant strains were inoculated into LB medium and egg white (final concentration 80%) and statically cultured at 42°C for 6 hours. RNA samples were then extracted for sequencing. Three biological replicates were carried out per sample. RNA sample was sequenced with Illumina HiSeq 2000 System. EdgeR method was used to calculate the differentially expressed genes. qRT–PCR and gene mutation were used to validate RNA-seq result. Results: A total of 12 samples were sequenced. The average clean bases of each sample was 3.63 Gb, and the average comparison rate with the reference genome was more than 90%. Compared with LB broth, there were 1606 differentially expressed genes (FDR ≤0.05 and |Log2Fold change| ≥1) in egg white with 762 genes upregulated and 844 genes downregulated. These accounted for about 35% of the whole genome indicating a large shift in the transcriptional response to egg white. qRT–PCR result of 7 selected genes highly consistent with the RNA-seq results. Five mutants showed decreased survival ability in egg white, suggested that the DEGs from the RNAseq results correlated significantly with the egg white resistance phenotype.Moreover, a significant downregulation of all genes within the fim gene cluster was observed and the RNA-seq analysis of Δfim mutant in the egg white was further conducted. In comparison with wild-type in egg white, the fim mutant showed 36 differentially expressed genes, with most of them associated with energy metabolism pathways.