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ARS Home » Southeast Area » Fayetteville, Arkansas » Poultry Production and Product Safety Research » Research » Publications at this Location » Publication #384440

Research Project: Multi-hurdle Approaches for Controlling Foodborne Pathogens in Poultry

Location: Poultry Production and Product Safety Research

Title: Sodium butyrate modulates chicken macrophages proteins essential for Salmonella Enteritidis invasion

Author
item GUPTA, ANAMIKA - University Of Arkansas
item BANSAL, MOHIT - University Of Arkansas
item LIYANAGE, ROHANA - University Of Arkansas
item UPADHYAY, ABHINAV - University Of Connecticut
item RATH, NARAYAN - Retired ARS Employee
item Donoghue, Ann - Annie
item SUN, XIAOLUN - University Of Arkansas

Submitted to: PLoS ONE
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/19/2021
Publication Date: 4/28/2021
Citation: Gupta, A., Bansal, M., Liyanage, R., Upadhyay, A., Rath, N., Donoghue, A.M., Sun, X. 2021. Sodium butyrate modulates chicken macrophages proteins essential for Salmonella Enteritidis invasion. PLoS ONE. 16(4):e0250296. https://doi.org/10.1371/journal.pone.0250296.
DOI: https://doi.org/10.1371/journal.pone.0250296

Interpretive Summary: Salmonella Enteritidis (S. Enteritidis) is a food-borne bacterial pathogen causing human gastrointestinal and systemic illness with limited prevention and treatment options. Butyrate is a short chain fatty acid derived from microbiota fermentation. In this study, chicken macrophages (HTC cells) were infected with S. Enteritidis, treated with sodium butyrate, and proteomic analysis was performed. A growth curve assay was conducted to determine sub-inhibitory concentration (SIC, concentration that do not affect bacterial growth compared to control) of sodium butyrate against S. Enteritidis. HTC cells were infected with S. Enteritidis in the presence and absence of SIC of sodium butyrate. The proteins of the cells were extracted and analyzed by tandem mass spectrometry. We found that the SIC for S. Enteritidis was 45 mM butyrate. Notably, S. Enteritidis-infected HTC cells were increased with proteins involved in ATP synthesis uch as ATP synthase subunit alpha (ATP5A1), ATP synthase subunit d, mitochondrial (ATP5PD) and cellular apoptosis such as Cytochrome-c (CYC). In addition, butyrate reduced the actin cytoskeletal rearrangement protein expression in S. Enteritidis-infected HTC cells such as WD repeat-containing protein-1 (WDR1), Alpha actinin-1 (ACTN1), Vinculin (VCL) and Protein disulfide isomerase (P4HB), as well as intracellular bacterial survival protein of V-type proton ATPase catalytic subunit A (ATPV1A). Interestingly, butyrate increased the expression of bacterial killing protein of Vimentin (VIM) in infected HTC cell. In conclusion, sodium butyrate modulates the expression of HTC cell proteins and the alteration may relate to S. Enteritidis invasion.

Technical Abstract: Salmonella Enteritidis is an intracellular foodborne pathogen that has developed multiple mechanisms to alter poultry intestinal physiology and infect the gut. Short chain fatty acid butyrate is derived from microbiota metabolic activities, and it maintains gut homeostasis. There is limited understanding on the interaction between S. Enteritidis infection, butyrate, and host intestinal response. To fill this knowledge gap, chicken macrophages (also known as HTC cells) were infected with S. Enteritidis, treated with sodium butyrate, and proteomic analysis was performed. A growth curve assay was conducted to determine sub-inhibitory concentration (SIC, concentration that do not affect bacterial growth compared to control) of sodium butyrate against S. Enteritidis. HTC cells were infected with S. Enteritidis in the presence and absence of SIC of sodium butyrate. The proteins were extracted and analyzed by tandem mass spectrometry. Our results showed that the SIC was 45 mM. Notably, S. Enteritidis-infected HTC cells upregulated macrophage proteins involved in ATP synthesis through oxidative phosphorylation such as ATP synthase subunit alpha (ATP5A1), ATP synthase subunit d, mitochondrial (ATP5PD) and cellular apoptosis such as Cytochrome-c (CYC). Furthermore, sodium butyrate influenced S. Enteritidis-infected HTC cells by reducing the expression of macrophage proteins mediating actin cytoskeletal rearrangements such as WD repeat-containing protein-1 (WDR1), Alpha actinin-1 (ACTN1), Vinculin (VCL) and Protein disulfide isomerase (P4HB) and intracellular S. Enteritidis growth and replication such as V-type proton ATPase catalytic subunit A (ATPV1A). Interestingly, sodium butyrate increased the expression of infected HTC cell protein involving in bacterial killing such as Vimentin (VIM). In conclusion, sodium butyrate modulates the expression of HTC cell proteins essential for S. Enteritidis invasion.