Elimination of Salmonella, Campylobacter, and spoilage bacteria in ground chicken patties through the application of Electron Beam (eBeam) Technology

Authors: PERERA, RUVINDU - University Of Arkansas; Jesudhasan, Palmy; ALRUBAYE, ADNAN - University Of Arkansas

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 4/28/2024
Publication Date: N/A
Citation: N/A

Interpretive Summary: Electron Beam (eBeam) technology is one of the best methods to develop safe and efficacious vaccine to animals, and humans. Developing vaccine is also a best alternative to antibiotics. The eBeam is generated using electricity, which is eco-friendly, cost-effective, devoid of health hazards, and rapidly provides targeted, high doses in a planar manner. Using eBeam, we can develop eBeam-killed vaccines. How eBeam inactivates pathogens is unique by shredding nucleic acids, so the cells can not replicate. The exposure of eBeam to bacteria does not affect epitopes or antigens, so the killed cells (vaccines)) are highly efficacious. The eBeam vaccines are arguably as safe as killed vaccines and as efficient as live vaccines. Salmonella is a major cause of non-typhoidal human salmonellosis, infecting approximately 1.5 million people in the US annually. Staphylococcus causes bacteremia and infective endocarditis, leading to osteoarticular, skin, soft tissue, pleuropulmonary, urinary tract, and device-related infections, and is a major human pathogen. Consumption of contaminated chicken is a major pathway for human infection by the above pathogens that also leads to the loss of revenue via condemnation of infected birds at the marketing age. With a long-term aim to produce vaccines against Staphylococcus and Salmonella infection in broiler chickens, this study strives to determine the optimum time to immunize birds based on retention of viability after eBeam treatment. We hypothesized that if birds are immunized while the bacterial cells are viable, the Pathogen Activated Molecular Patterns produced by the organisms may contribute to immunogenicity. Cultures of both bacteria (~108 CFU/mL) were inactivated at a range of high energy eBeam doses (4.5 through 9kGy), previously known for complete inactivation, and their viability was evaluated (using Promega - BacTiter-Glo™ Microbial Cell Viability Assay) weekly over 24 days. This assay quantifies the amount of Oxygen and ATP in viable cells by oxygenating Beetle Luciferin to luminescent Oxyluciferin by a luciferase enzyme, which then outputs the viability as relative luminescence. Each sample/time point viability was measured in triplicate and compared to a fresh bacterial sample. Brown-Forsythe and Welch ANOVA, with Dunnett’s T3 multiple means comparison, was conducted to compare the viability of samples/time points at the 95% confidence level. The viability of Salmonella was triggered throughout, while Staphylococcus significantly dropped viability by the 3rd week after eBeam exposure, irrespective of the dose. This test concluded that an eBeam-killed-Salmonella vaccine is usable even after a month, but an eBeam-killed-Staphylococcus vaccine may be used for immunization within its first week.

Technical Abstract: Electron Beam (eBeam) treatment is among the most successful options to produce safe and effective vaccines against poultry pathogens as an alternative to antibiotics. The source of eBeam is electricity, which is eco-friendly, cost-effective, devoid of health hazards, and rapidly provides targeted, high doses in a planar manner. eBeam shreds nucleic acids (restricting reproduction) and retains the membrane integrity of exposed bacteria, thus preserving antigenic epitopes and immunogenicity. So, eBeam vaccines are arguably as safe as killed vaccines and as efficient as live vaccines. Salmonella is a major cause of non-typhoidal human salmonellosis, infecting approximately 1.5 million people in the US annually. Staphylococcus causes bacteremia and infective endocarditis, leading to osteoarticular, skin, soft tissue, pleuropulmonary, urinary tract, and device-related infections, and is a major human pathogen. Consumption of contaminated chicken is a major pathway for human infection by the above pathogens that also leads to the loss of revenue via condemnation of infected birds at the marketing age. With a long-term aim to produce vaccines against Staphylococcus and Salmonella infection in broiler chickens, this study strives to determine the optimum time to immunize birds based on retention of viability after eBeam treatment. We hypothesized that if birds are immunized while the bacterial cells are viable, the Pathogen Activated Molecular Patterns produced by the organisms may contribute to immunogenicity. Cultures of both bacteria (~108 CFU/mL) were inactivated at a range of high energy eBeam doses (4.5 through 9kGy), previously known for complete inactivation, and their viability was evaluated (using Promega - BacTiter-Glo™ Microbial Cell Viability Assay) weekly over 24 days. This assay quantifies the amount of Oxygen and ATP in viable cells by oxygenating Beetle Luciferin to luminescent Oxyluciferin by a luciferase enzyme, which then outputs the viability as relative luminescence. Each sample/time point viability was measured in triplicate and compared to a fresh bacterial sample. Brown-Forsythe and Welch ANOVA, with Dunnett’s T3 multiple means comparison, was conducted to compare the viability of samples/time points at the 95% confidence level. The viability of Salmonella was triggered throughout, while Staphylococcus significantly dropped viability by the 3rd week after eBeam exposure, irrespective of the dose. This test concluded that an eBeam-killed-Salmonella vaccine is usable even after a month, but an eBeam-killed-Staphylococcus vaccine may be used for immunization within its first week. A complete understanding of the effect of viability on immunogenicity warrants further follow-up studies.