Researcher(s)
- Violet Geary, Animal Science, University of Delaware
Faculty Mentor(s)
- Alexander Yitbarek, Animal and Food Sciences, University of Delaware
Abstract
Clostridium perfringens (c. perfringens) is a bacteria in the gastrointestinal tract that is known to be one of the lead causes for necrotic enteritis (NE) in chickens. In this experiment, the pathogenicity of c. perfringens at different iron doses in chicken embryos was evaluated to determine whether a high iron concentration could be linked to NE. This was the first time to our knowledge that C. perfringens was tested using chicken embryos. C. perfringens was grown in a Brain Heart Infusion broth (BHI) media. Treatments included a PBS control, only C. perfringens challenge, C. perfringens + 10ug iron sulfate solution in PBS, and C. perfringens + 30ug iron sulfate solution in PBS. Chicken embryos were challenged on day 17 with 10^8 CFU of C. perfringens and necropsies were performed on day 19 to collect samples from the ileum, spleen, and bursa of Fabricius. mRNA was extracted and nucleic acid concentration (NAC), 260/280, and 260/230 purity levels were determined in a Nanodrop machine. To ensure credible mRNA reliability, mRNA purification performed and re-measured NAC, 260/280, and 260/230 in the Nanodrop machine once more. Once mRNA levels were satisfactory, cDNA synthesis was performed followed by gene expression analysis using quantitative real-time PCR to measure the CT values of samples. Finally, the fold expression of IL-1Beta, IL-10 and IL-22 was calculated using relative quantification. Data were analysed using ANOVA in R (R Studio) and a P < 0.05 was considered statistically significant. Results indicated that chicken embryos may be a useful model in studying host-pathogen interactions and assessment of intervention strategies against necrotic enteritis, and that Fe with C. perfrigens was associated with high pro-inflammatory markers in the ileum and spleen.