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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - PHAGOVET (A cost-effective solution for controlling Salmonella and Escherichia coli in poultry production)

Teaser

Summary

The Phagovet project is devised to satisfy market demand for a more secure and reliable solution to control Salmonella and E. coli infections in poultry farms. To date, avian colibacillosis and salmonellosis are considered the main bacterial infections in the poultry sector having an important economic impact worldwide. In addition, among other causes, the overuse of antibiotic in farming has caused a rise in antimicrobial resistance 700,000 people die annually from drug resistant infections and it is estimated that this number will rise to 10 million by 2050. In response to the unmet need, Phagovet proposes a cost-effective alternative to antibiotics against Salmonella and E. coli. Phagovet consists of a biocide and two technological food additive products based on selected bacteriophages able to kill the target bacteria. Phagovet will be the first phage-based solution to naturally and effectively control both bacteria in poultry production while avoiding side effects and residues associated to antibiotic use. It will reduce Sallmonella infections by 20% and reduce the use of antibiotics on E. coli by 50%. Thus, Phagovet provides a promising effective control measure for a concerning problem for poultry farmers and for human health

Work performed

The main objective of the tasks performed in this period was to define the best phage composition of Phagovet products, which must accomplish both criteria of wide range of lytic activity and safety. For this, phages specific against E. coli have been isolated and their lysis efficiency were evaluated in vitro by spotting method in a large collection of E. coli strains isolated from European poultry industries. Seven phages, which presented the highest lysis efficiencies, were selected for further studies. On the one hand, they were morphological characterized by TEM. Taxonomic analysis showed that all of them belong to Caudovirales order, one is a member of the Podoviridae family and the others, to the Myoviridae family. On the other hand, genetic analysis were performed. RAPD-PCR assays were carried out to assess genetic diversity between the phages. The results obtained showed different band patterns from 6 phages. Also, we have already sequenced the genome of four phages. Preliminary analysis did not revealed any similarities neither with known virulence-associated genes nor with any antibiotic resistance genes. Genes commonly associated with lysogeny were not found in two phages, which also showed high similarity with lytic bacteriophages. However, the others two phages apparently have putative genes associated with genome integration abilities. Nevertheless, further studies are mandatory to confirm if theses phages are lysogenic and/or have transduction capability. Lysogen formation assays are being optimized and carried out.
In view of the scaling-up phage production, the growth parameters of the phages were evaluated by One-Step Growth Curve experiments. Preliminary analysis showed that the latent period of the phages was of 15-20 minutes, while their average burst size was highly variable.
The E. coli strains tested for phage lytic activity, were also examined by PCR for the presence of 8 virulence factors associated with colibacillosis. Different combinations of selected genes were found among the strains. 84.0% of the isolates were positive for at least one of the genes

Final results

Nowadays, antibiotics have been routinely used in poultry to promote animal growth or prevent disease outbreaks. However, among other causes, their overuse has led to drug-resistant strains, limiting its therapeutic effectiveness and increasing antimicrobial resistances, being then a major problem, both in animal and human medicine. What is shocking, is that in Europe 3 times more antibiotics are used on livestock rather on people due to routine mass medication. Development of new antibiotics is difficult and slow, with a recent discovery of new antibiotic class after 30 years of no breakthrough. Thus, because of the serious risk to public health, in 2017 the American Food and Drug Administration (FDA) has limited the use of antibiotics only for treating or preventing disease only with veterinary prescriptions. In 2006 the European Union implemented a ban on antibiotics as growth promoters and will implement new restrictive legislation in 2018. These are likely to follow FDA legislation. Finally, consumers are growing more aware of the problem and choose antibiotic-free products.
Phagovet is an innovative solution based on patented technology of phagovet cocktail and proprietary processes combined with years of lab research and experience in the microbiology field, we will produce the first variety of bacteriophage-based products for the effective control of both E. coli and Salmonella in poultry farms.
The project will tap into the global Animal Healthcare market which was worth â‚¬28.4bn in 2016 and is growing at 5.1% CAGR.17 As a consequence, we expect to reach â‚¬17.76M profit at the end of the 2026 as the project will open the European and, most importantly, the global phage market. In this way, not only will it strengthen our industrial partners and boost the investigation capabilities of our R&D partners, but also have a significant long-term impact on EU economy and its employment.
Apart from financial success, our goal is to provide our clients, poultry farmers, an efficient, cost-effective, easy-to use and safe tool to combat E. coli and Salmonella, an alternative or compliment to the currently used antibiotics which are gradually phased out due to the concern with antimicrobial resistance (AMR) rise. Consequently, bacteriophages are currently the most promising answer to this global health and safety problem and Phagovet will be a major step towards an effective solution. As a result, we will significantly enhance the EU health safety