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

通过这个项目，我们提出了两项创新技术，用于阻止AMR病原体在家禽链中的传播，即来自加拿大的羟基自由基技术和来自英国的噬菌体生物杀菌技术。虽然水基羟基自由基体系被广泛使用，但在气相中的应用是一个相对较新的发展，因此需要更深入地研究气相版本对食品及其环境的有效性。家禽链有效性将解决家禽链中的抗菌素耐药性病原体。英国诺丁汉大学的已故威廉·韦茨教授是第一个看到气相高级氧化工艺（AOP）在食品加工中的潜力的人。气相-羟基自由基过程通过紫外光介导的过氧化氢和臭氧降解产生高度抗菌的蒸汽。自由基灭活微生物而不留下有毒残留物。该技术是灵活的，可以应用于隧道，批量系统或手持设备的形式。在这个项目中，我们将使用羟基自由基对家禽环境、鸡蛋、板条箱、禽肉等进行消毒。羟基自由基处理可有效灭活病原菌，但无残留抗菌活性。因此，应用噬菌体后羟基自由基治疗可防止病原体在消毒表面重新建立。我们的加拿大合作伙伴的研究已经证明了AOP在多种水果和蔬菜类型上的有效去污，并且具有延长保质期的额外好处。他们目前的研究应用了同样的方法来净化去壳鸡蛋、板条箱和禽肉。在孵化研究中，已经证明羟基自由基过程可以在10s内灭活沙门氏菌（减少5 log CFU），而不影响鸡蛋的完整性或胚胎的发育。噬菌体（简称噬菌体）是自然产生的细菌病毒，它专门感染并杀死细菌，只留下有益的微生物。噬菌体的这种能力被用来控制各种环境下的细菌。环境应用的主要问题是应用方法和噬菌体的生存能力，特别是尾部噬菌体。本项目将探索通过干燥的噬菌体粉末，在现场应用时将其溶解在水中，从而实现可持续的噬菌体应用方法，并对尾状噬菌体与非尾状噬菌体的生存能力进行比较。此外，沙门氏菌和弯曲杆菌的目标菌株在英国和加拿大是最普遍的，这将有利于这两个国家的家禽连锁店。我们的干噬菌体粉末方法的新颖之处在于开发了尖端的原型稳定噬菌体产品，可以轻松廉价地加入水中进行环境喷洒，或直接应用于动物尸体以去除沙门氏菌和弯曲杆菌。这一点很重要，因为传统的噬菌体产品不稳定，难以输送给动物或应用于肉类产品。因此，使用噬菌体的潜在好处被这些将科学转化为可行的商业产品所需的障碍所掩盖。使用喷雾干燥噬菌体的原因是该技术具有高度可扩展性，广泛应用，高效和廉价的方法。稳定的噬菌体产品不需要复杂的储存，它消除了输送的障碍。这种方法生产粉末状噬菌体的可行性已经在评估加工成粉末的潜力的研究中得到了证明。我们将整合这两种技术，其中羟基自由基技术（基于蒸汽的技术，能够提供包括空气消毒在内的整体体积）和噬菌体生物杀菌剂，以控制将雏鸡重新引入环境中的再污染。

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.