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病原体在家禽产业链中的传播。来自加拿大的羟基自由基技术，以及来自英国的噬菌体生物香料技术。尽管水基羟基自由基体系被频繁使用，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在食品及其环境中的有效性。家禽链的有效性将解决家禽链中的AMR病原体。英国诺丁汉大学已故教授威廉·韦茨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-羟基自由基法通过紫外光介导的过氧化氢和臭氧的降解产生高度抗微生物的蒸气。这些自由基能灭活微生物，不会留下有毒残留物。该技术灵活，可以隧道、批量系统或手持设备的形式应用。在本项目中，我们将使用羟基自由基对禽类环境、鸡蛋、板条箱、禽肉等进行消毒。羟基自由基处理可以有效地灭活病原体，但没有残留的抗菌活性。因此，噬菌体后羟基自由基处理的应用将起到防止病原体在消毒表面重新建立的作用。我们的加拿大合作伙伴的研究表明，AOP在各种水果和蔬菜类型上都能有效地去污，而且还能延长货架寿命。他们目前的研究也应用了同样的方法来净化去壳的鸡蛋、板条箱和禽肉。在孵化场的研究中，已经证明羟基自由基过程可以在10s(5logCFU减少量)内灭活沙门氏菌，而不会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，它们专门感染和杀死细菌，而不是有益的微生物。噬菌体的这种能力正在被利用来控制各种环境中的细菌。环境应用的主要关注点是噬菌体的使用方法和生存能力，尤其是尾噬菌体。本项目将探索通过干噬菌体粉剂进行可持续的噬菌体应用的方法，并检测尾噬菌体与非尾噬菌体的活性。此外，将针对的沙门氏菌和弯曲杆菌是英国和加拿大最流行的菌株，这将有利于这两个国家的家禽连锁店。我们的干噬菌体粉法的创新之处在于开发了尖端原型稳定的噬菌体产品，这些产品可以轻松且廉价地掺入水中进行环境喷洒，或直接应用于动物身体以去除沙门氏菌和弯曲杆菌。这一点很重要，因为传统上噬菌体产品不稳定，很难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将科学转化为可行的商业产品所需的这些障碍所掩盖。之所以使用喷雾干燥噬菌体，是因为该技术是一种高度可扩展、广泛使用、高效和廉价的方法。稳定的噬菌体产品消除了复杂储存的需要，并消除了运送障碍。这种方法生产噬菌体粉末的可行性已经在评估加工成粉末的潜力的研究中得到证明。我们将整合这两项技术，其中羟基自由基技术(基于蒸气的技术能够提供包括空气消毒在内的整个体积)和噬菌体生物消毒剂来控制重新引入孵化日小鸡到环境中的再次污染。

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.