Effect of electron beam irradiation on the immunosuppressive properties of deoxynivalenol in HD11 chicken macrophage cells following lipopolysaccharide challenge

Authors: KAPPARI, LAHARIKA - University Of Georgia; Jesudhasan, Palmy; SELVARAJ, RAMESH - University Of Georgia; APPLEGATE, TODD - University Of Georgia; Shanmugasundaram, Revathi

Submitted to: Poultry Science Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 4/23/2025
Publication Date: N/A
Citation: N/A

Interpretive Summary: N/A

Technical Abstract: Deoxynivalenol (DON), a prevalent mycotoxin in poultry feed, adversely impact chicken health due to its immunosuppressive effect and results in production losses. Electron beam irradiation (EBI) is widely used for the decontamination of microorganisms in feedstuff. Hence, our objective was to assess the efficacy of EBI to denature DON. In vitro testing was used to determine the optimal dose in kilogray (kGy) that can effectively denature DON using chicken HD11 macrophage cell line. We hypothesized that increasing exposure of EBI from 2.5 to 30 kGy would denature the DON as a result of reduced immunosuppressive effects and restore nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated HD11 chicken macrophage cells comparable to the control + LPS group. HD11 cells at 1x105 cells/mL concentration were plated with 0 or 1'g/mL of DON and EB irradiated DON (2.5, 5, 10, 15, 20, and 30 kGy), followed by stimulation with 10 'g/mL lipopolysaccharide (LPS). After 48h of incubation, the supernatant was used to measure NO production, and real-time qPCR was performed to analyze the Toll-like receptor (TLR)-2, TLR-4, and inducible nitric oxide synthase gene expression (iNOS) (n = 3). The statistical analysis was done using one-way ANOVA, and means were separated using the Tukey HSD test. The non-irradiated DON with the LPS challenge significantly reduced the nitrite concentration by 54% (p < 0.05). Similarly, the EBI treated DON with LPS challenge at doses of 2.5-5 kGy, 10-15 kGy, and 20-30 kGy significantly reduced nitrite concentration by 53-58%, 83-86%, and 76%, respectively, when compared to the control + LPS challenge group (p < 0.05). Gene expression analysis results showed that there is no significant difference in TLR-4 expression (p > 0.05), whereas TLR-2 expression was significantly downregulated in both nonirradiated and irradiated treatment groups (p < 0.05) when compared to the control group, indicating the immunosuppressive effect of toxin even after irradiation. There was a significant reduction in inducible nitric oxide synthase (iNOS) expression by 0.35-fold in the non-irradiated DON group with the LPS challenge (p < 0.05). Similarly, there was a significant reduction in iNOS expression in irradiated treatment groups when compared to control+LPS (p < 0.05). These results indicate that EBI up to 30 kGy did not mitigate the negative impact of DON on HD11 cells, likely due to the formation of additional toxic metabolites resulting from EBI that induce negative effects. In order to verify this, HPLC can be used to quantify DON metabolites in the cell culture and can be done in future experiments. In conclusion, EBI is ineffective for pre-formed toxins but may be useful for decontaminating fungi in feedstuffs. Keywords: Electron beam irradiation; Fumonisin; Deoxynivalenol; Lipopolysaccharide.