Canada_IPAP: Application of Hydroxyl radical process and phage biosanitizers to reduce the spread of AMR pathogens in the poultry chain.

Through this project we are proposing two innovative technologies to be used to stop the spread of AMR pathogens in poultry chain viz. Hydroxyl radicals' technology from Canada, and a phage biosanitizer technology from the UK. Although aqueous-based hydroxyl radical systems are used frequently, the application in the gas phase is a relatively new development and hence more in-depth studies on the effectiveness of gas phase version on food commodities and their environments is needed. Poultry chain effectiveness will address AMR pathogens in the poultry chain. Late Prof William Waites of the University of Nottingham, UK was the first to see the potential of gas phase Advanced Oxidation Process (AOP) in food processing. The gas phase-hydroxyl radical process generates highly antimicrobial vapour through the ultraviolet light mediated degradation of hydrogen peroxide and ozone. The radicals inactivate microbes without leaving toxic residues. The technology is flexible and can be applied in the form of a tunnel, batch system or handheld device. In this project we will use Hydroxyl radicals to disinfect poultry environments, eggs, crates, poultry meat etc. The hydroxyl-radical treatment can effectively inactivate pathogens although there is no residual antimicrobial activity. Therefore, the application of bacteriophage post-hydroxyl radical treatment will act to prevent pathogens becoming re-established on the disinfected surface. Research by our Canadian partner has demonstrated effective AOP decontamination over a diverse range of fruit and vegetable types with an added benefit of extending shelf-life. Their current research has applied the same method for decontaminating shelled eggs, crates and poultry meat. Within the hatchery studies it has been demonstrated that the hydroxyl radical process can inactivate of Salmonella within 10s (5 log CFU reduction) without effecting the egg integrity or embryo development.

Bacteriophages (phages in short) are naturally occurring bacterial viruses which specifically infect and kill bacteria leaving good microbes alone. This ability of the phages is being harnessed in controlling bacteria in various settings. The major concern for environmental application is method of application and viability of phages especially of the tailed phages. This project will explore the sustainable method of phage application through dry phage powder which could be dissolved into water during field application and also check the viability of tailed phages compared to non-tailed phages. Moreover, the strains of Salmonella and Campylobacter that will be targeted are the most prevalent in the UK and Canada which will be beneficial to the poultry chains in both the countries. The novelty of our dry phage powder approach lies in the development of cutting-edge prototype stable phage products that can be easily and cheaply incorporated into water for environmental spraying, or applied directly to animal carcasses to remove Salmonella and Campylobacter spp. This is important because traditionally phage products are unstable, and difficult to deliver to animals or applied to meat products, and thus potential benefits of using phages have been overshadowed by these hurdles needed to translate the science into a viable commercial product. The reason for using spray dried phages is that the technique is a highly scalable, widely used, efficient and inexpensive method. A stable phage product negates the need for complicated storage and it removes barriers for delivery. The feasibility of this method to produce powdered phages has already been proven in studies assessing the potential of processing into powders.

We will integrate these two technologies wherein Hydroxyl radical technology (vapour based which is able to provide whole volume including air disinfection) and phage biosanitizers to control recontamination from re-introduction of day-of-hatch chicks into the environment would be possible.

Two innovative technologies viz. hydroxyl ion disinfection and phage dry powder as a biosanitizer are core part of this collaborative project. We would put efforts to bring clear synergy between these two technologies to tackle AMR in poultry chain.

Although aqueous-based hydroxyl radical systems are used frequently, the application in the gas phase is a relatively new development and hence more in-depth studies on the effectiveness of gas phase version on food commodities and their environments is needed. Poultry chain effectiveness will address AMR pathogens in the poultry chain. This process is based on the UV mediated degradation of hydrogen peroxide and ozone gas generating antimicrobial vapour. Studies with the hydroxyl-radical treatment will focus on engineering designs and operating parameters to be applied to the different settings of the poultry chain. This will enable the design of the gas-phase advanced oxidation process unit by our Canadian industrial partner to be validated in field trials.

We will also use dry powder phage biosanitizers to specifically tackle strategically important bacteria in poultry environment such as Salmonella spp and Campylobacter spp. Mainly, optimise formulation (determine suitable excipients) and spray drying parameters to minimise phage loss during spray drying and upon long term storage at both refrigerated and ambient conditions. Prior to this, the most active phages against a selection of Salmonella and Campylobacter isolates that are most prevalent in the UK and Canada would be characterized biologically, physically and genomically. Cocktail of phages will be formulated for broad spectrum activity and then field studies would be carried for evaluating the efficacy (of tailed Vs non-tailed phages) in poultry chain by spraying phages after dissolving dry phage powder into water.

This is a translational proof-of-concept study that combines above two novel approaches to control AMR pathogens in poultry chain.