Understanding of Phage Technology Growing by Leaps and Bounds, Unlocking Benefits for Animal Production – INDUSTRY PERSPECTIVES

In the context of an alarming escalation in antibiotic resistant strains of bacteria and a dwindling pipeline for new antibiotics, it has become clear to animal producers around the world that alternative solutions to prevent and treat bacterial infections in animals are urgently needed, allowing us to keep antibiotics working for when they are most needed.  

One of the most exciting antibiotic alternative solutions is the category of bacteriophages – natural self-replicating microscopic organisms which target specific pathogenic bacteria without disrupting the host or beneficial microbiome. While a basic understanding of phages has existed since the early 20^th century, the technological innovations in recent years have transformed our ability to understand them, unlocking a tool that can be reliably used to keep bacterial diseases in check.

With its 18 years of experience in phage technology, Poland’s Proteon Pharmaceuticals considers itself a leader in veterinary phages. Today, we hear from the company’s Chief Product and Technology Officer, Justyna Andrysiak about the breakthroughs in R&D, particularly over the last year, and how it will continue to push forward the frontiers of phage research in the future.

[Feedinfo] Science has known about phages for over a century, but in recent years, there have been technological developments that have allowed us to better understand and more consistently produce them. Can you talk about that shift in the technological landscape?

[Justyna Andrysiak] Indeed, phages were initially discovered over a century ago. However, back then, our scientific capabilities were significantly constrained, and we lacked the essential tools for distinguishing between virulent and lysogenic (temperate) phages. This limitation was primarily due to the absence of the molecular biology techniques, advanced sequencing technology, and bioinformatics resources that we now possess. Now, we know that lysogenic phages remain dormant for a certain period. As a result, when these phages were used in treatments, their presence could potentially compromise effectiveness. At Proteon, we exclusively employ virulent phages. Their singular life cycle ensures the rapid destruction of bacterial cells. Presently, we have a comprehensive toolkit for molecularly identifying and characterizing phages. We can analyze their genomes at the protein level and model their structures. Additionally, we utilize AI tools such as the PhageAI application to analyze their life cycle and immediately classify them as virulent or temperate. This guarantees absolute confidence in achieving the desired antibacterial effect when incorporating only virulent phages into our products.

[Feedinfo] Proteon has 18 years of experience in phages for animal production. How long have its products been commercially available in different markets? How has this experience improved your understanding of phages, as well as the quality of the final products?

[Justyna Andrysiak] First and foremost, I'd like to emphasize the remarkable transformation in the regulatory landscape over the past 18 years. When we embarked on this journey, few would have wagered that we could successfully bring a phage-based product to market. Yet, here we stand, as one of the pioneering phage companies with commercially available products. Our initial market entry was in India, where we've secured registrations for all our products. Furthermore, we've obtained registration approvals for our Salmonella product, BAFASAL, in Brazil and the USA, with an ongoing and highly advanced registration process in the European Union.

I want to highlight that being among the first pioneers to introduce phage products in various markets has been an invaluable experience for us. First of all, we feel that we had an influence in shaping the standards for testing phage preparations through our proactive engagement in discussions with different regulatory agencies. Secondly, we are sincerely appreciative of our clients' feedback, which has played a pivotal role in refining our products, making them more and more practical and user-friendly. We strive to meet our clients' expectations while maintaining an unwavering commitment to the quality of our products.

[Feedinfo] One of your major priorities in 2023 was testing products for shrimp. Does the aquatic environment impose any unique challenges when it comes to the delivery of a phage-based feed additive? What can you tell me about the details of the trials, and their results?

[Justyna Andrysiak] At Proteon, we fully understand the crucial role aquaculture plays in sustainable seafood production, alleviating overfishing pressures on natural populations, ensuring food security, and fostering economic growth. Indeed, the aquatic environment differs significantly from the terrestrial one in many aspects. Consequently, a dedicated effort in our research and development is applied to customize phage products to meet the distinctive requirements of the aquatic environment. This process consists of enhancing phage stability, devising efficient delivery mechanisms, and ensuring alignment with the specific conditions of the aquaculture system.

Since 2021, we've been working on a phage solution designed to prevent Vibrio parahaemolyticus infections in shrimp. Our strong partnership with a major player in the industry provides the expertise and practical knowledge needed to tailor products to meet the specific needs of our clients. Currently, our prototype is prepared, and we have conducted recent proof-of-concept testing in shrimp. While I'm unable to disclose further details at this juncture, I can confirm that the outcomes have been highly encouraging and positive.

[Feedinfo] Beyond this product-specific development work, you have also made significant progress in general-application bacteriophage research. What were some of the most important findings or accomplishments here?

[Justyna Andrysiak] Our research has truly been innovative in several key aspects. To name a few, we've successfully developed the capability to incorporate phages into various formats, ranging from microcapsules to powders, gels, and hydrogels, all while preserving their high bioactivity.

Additionally, we've constructed complex in vitro models that faithfully mimic the conditions we aim to test. The controlled environments provided by in vitro models have been instrumental in advancing our product development. They have enabled us to meticulously manipulate specific variables, allowing for a deeper understanding of the intricate interactions related to phages in a reproductive context. Moreover, they have significantly reduced the necessity for experiments on living organisms during the initial stages of our product development process. This not only accelerates the development timeline but also aligns with ethical considerations and safety concerns associated with live organism experiments.

We have recently implemented our High-Throughput Screening (HTS) methodology to efficiently isolate and characterize new bacteriophages. HTS allows us to work with more demanding bacteria and significantly speeds up the entire isolation process.  For the purpose of deep characterization of phages we are utilizing the PhageAI tool, an application that serves as both a repository of knowledge about bacteriophages and a tool that employs Artificial Intelligence for genome analysis.  

[Feedinfo] What should the industry be expecting from Proteon Pharmaceuticals in 2024?

[Justyna Andrysiak] Proteon is a pioneering company known for its bold initiatives. We proudly claim the title of being the first company to successfully apply for the registration of phage products in various markets. Additionally, we are at the forefront of the registration process for a phage-based feed additive with the European Food Safety Authority in the European Union, setting the benchmark for future applications in this domain. As an example for this process, we are one of the first laboratories in Europe or maybe even in the world to have successfully validated and implemented a quantitative method for assessing the biological activity of phages using PFU (Plaque Forming Units) enumeration in accordance with Good Laboratory Practices (GLP) standards. This approach offers an objective means of quantifying phages in solutions, while also factoring in their biological activity, in contrast to PCR-based tests. Notably, this method has received approval from European Reference Laboratory as a trustworthy and reliable assessment technique.

Presently, we are actively seeking new partners who are eager to broaden their product portfolio within the realm of phage products. Proteon stands as a dependable ally in this endeavour, demonstrating a strong commitment to the development of innovative antibacterial products.

In 2024, we are poised to inaugurate our state-of-the-art factory in India, showcasing our commitment to expanding our global presence. One of our core strengths lies in our adaptability, which drives us to keenly observe market trends and tailor our product portfolio to meet the evolving needs of our customers.

Published in association with Proteon Pharmaceuticals