The development of a phage food additive with the aim to control Salmonella in swine and poultry

Lead Research Organisation: University of Leicester
Department Name: Infection Immunity and Inflammation

Abstract

Animal experimental design to study the role of phage therapy in acute-non typhoidal Salmonellosis
Co-investigator: Dr. Parameth Thiennimitr, M.D.,Ph.D.

Address: Department of Microbiology, Faculty of Medicine, Chiang Mai University

Species of animal: 6-8 weeks old female mouse (Mus musculus) C57BL6 strain
Protocol (in brief): C57BL/6 mice will be purchased from Nomura Siam International. Mice will be acclimatized for at least 1 week before the experiment day. 24 hour before Salmonella enterica Typhimurium (strain IR715) infection, mouse will be orally gavaged with 20 mg streptomycin sulphate to allow subsequent STM IR715 to establish a colitis. Streptomycin-treated mice will be orally infected with 100 microlitr of 109 cfu/ml STM IR715 solution. Then, 100 microlitr of Salmonella phage solution will be orally gavaged to the infected mice for consecutively 3 days. On day 4 post infection, mice will be euthanized by CO2 and cervical dislocation. Mouse colon content, colon, spleen will be collected for further analysis. Control group (no treatment) of mice will be orally fed with normal saline solution as used as a vehicle for the experimental (phage therapy) groups.
Animal protocol approval committee: All animal experiments conduct at Chiang Mai University will be approved by the Chiang Mai University Animal Care and Use Committee (CMU-ACUC). All animal experiments will be performed in the AAALAC-certified biosafety level (BSL) 2 facility located in Chiang Mai University. Numbers of animal in each group is 7. [Calculated followed Bernard, R. (2000). Fundamentals of biostatistics (5th ed). Duxbery: Thomson learning, 308, level of significance = 0.05)]

Planned Impact

Infections caused by antibiotic resistant bacteria currently result in more than 50,000 deaths annually across Europe and the US, with many more people dying worldwide. The emergence of bacterial resistance to all classes of antibiotics is a major global health concern, with the number of attributable deaths worldwide predicted to reach 10 million by 2050. The associated economic, and social loss highlights the pressing need to develop alternative therapeutics. As the identification and development of new antibiotics is slow and difficult, there is an urgent need to explore other viable alternatives. Our work is centred on developing a novel phage based therapeutic approach for Salmonella, which is an important problem worldwide. It is increasingly becoming resistant to antibiotics and both single and multidrug resistant strains have been isolated from the swine and poultry in the farm environment. There is therefore a particular incentive to identify novel therapeutics to prevent and treat infection. Although Salmonella is our focus, the novel timely approaches and tools we will develop will be widely adaptable to target other bacterial gastrointestinal pathogens.

Harnessing the lytic activity and specificity of phages offers a plausible alternative approach to treatment of MDR bacteria, and investment in development of this technology offers an opportunity to have a significant impact in the medium to long term on the quality of life and health. Whilst in the short term, our research will reduce Salmonella from a farm environment and prevent it getting into the human food chain, the longer-term future beneficiaries of this research will be much wider. Clearly the work will have a huge benefit to the export market as currently chicken meat is not allowed to be exported to the EU if it is found to be contaminated with Salmonella. The work will also benefit healthcare providers in Thailand and the UK, and reduce the amount of time nationally that is lost to illness caused by Salmonella infection and subsequent gastrointestinal disorders known as Salmonellosis. Other beneficiaries of the research include the veterinary industry and farmers who are under increasing pressure to not prescribe antibiotics but yet need to treat their livestock.

The proposed research will contribute to advances in knowledge and understanding of how phages can be optimally developed will be used to confirm they have increased efficacy. Again in a context wider than Salmonella, this work will improve efficiency of formulations, and the effective quantitative delivery of phages that target enteric diseases. The beneficiaries of this research include academic and industrial scientists working in the field of biotechnology, and specialists in drug delivery who will benefit from our novel methods and approaches. The lack of methods to allow safe delivery of biological products in the gastrointestinal tract currently limits delivery of all biological therapeutics and our encapsulation technology could also be applied to other orally delivered, biologically based drug and vaccine candidates.

To summarise, our efforts to construct a stable phage preparation will be of benefit to food producers in Thailand and the UK, and in academic and industrial researchers working on developing both prophylactic and therapeutic treatments for a wide range of gastrointestinal infections. Whilst our target country is Thailand, we could in future work extend the phage product development and make it accessible to swine and chicken producers in China, the Philippians and further afield.

Publications

10 25 50
 
Description We have identified a phage product that targets relevant strains of Salmonella associated with swine and poultry. We have shown the phages are stable during the drying progress and we are working on formulation.
Exploitation Route Our findings are currently being published. We are also exploring relationships with companies.
Sectors Agriculture, Food and Drink

 
Description We are working with a company to develop these phaeges commercially.
First Year Of Impact 2016
Sector Agriculture, Food and Drink,Healthcare
Impact Types Societal,Economic

 
Description New collaboration to phage efficacy in chickens 
Organisation University of Liverpool
Country United Kingdom 
Sector Academic/University 
PI Contribution We are developing a phage based product to remove Salmonella infection from chickens
Collaborator Contribution Prof Paul Wigley from the University of Liverpool will work with us to test the phages in his chicken model
Impact We will do the trail this year
Start Year 2019
 
Title Therapeutic phages 
Description The phages all target serovars of Salmonella associated with UK pigs and poultry. They have broad host range and are thermally stable so useful for downstream properties. 
IP Reference PCT/GB20 1815483.1 
Protection Patent application published
Year Protection Granted 2018
Licensed Commercial In Confidence
Impact Currently under negotiation.
 
Title We have carried out an efficacy trial on 750 chickens 
Description We worked with a major feed producer in order to determine how phages may be developed for use in poultry - they are currently assessing the data 
Type Therapeutic Intervention - Drug
Current Stage Of Development Initial development
Year Development Stage Completed 2020
Development Status Actively seeking support
Impact Currently investigating further 
 
Description ACTACC Annual Scientific Meeting 2021 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Gave plenary talk to UK heart and lung anaesthetists at their annual meeting in Glasgow
Year(s) Of Engagement Activity 2022
URL https://www.actacc.org/meetings-and-events
 
Description BBC R4 Life Scientific 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact This had huge impact I have developed many collaborations on the back of it and reached out to teachers and clinical researchers
Year(s) Of Engagement Activity 2019
URL https://www.bbc.co.uk/programmes/m0009b2t
 
Description BSAC/GARDP/IRAADD/DZIF Webinar Conference: Antimicrobial Chemotherapy, 2-3 February 2021 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Conference lots of questions asked and further invitations for discussions
Year(s) Of Engagement Activity 2022
 
Description CF pharmacist group meeting (Douglas McCabe - Scottish NHS) virtual 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact October 2021, phage therapy to treat MDR NTM infections in our patients with cystic fibrosis
Year(s) Of Engagement Activity 2022
 
Description Centenary Celebrations: TED Talks 2-Oct-2021 Not all viruses are bad; how bacterial viruses could help save us from the next pandemic Talk time TBC - between 11.00-16.00 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Keynote presentation
Year(s) Of Engagement Activity 2022
 
Description Department of Evolutionary and Environmental Biology, University of Haifa 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Seminar
Year(s) Of Engagement Activity 2022
 
Description IV Paranaense Congress of Microbiology - Brazil (virtual) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Talk to Brazilian scientists
Year(s) Of Engagement Activity 2022
 
Description Medical Biodefense Conference (Munich) (Hybrid virtual/in person) 28 Sept - 1 Oct 2021 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Talk to German biodefence organisation
Year(s) Of Engagement Activity 2022
 
Description Office of Biodefense, Research Resources and Translational Research/DMID/NIAID (Joe Campbell), USA on animal phage studies informing human trials 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact N/A
Year(s) Of Engagement Activity 2022
 
Description Phage Ecology Informing Applied Development, University of Nottingham (Schools of Biosciences and Veterinary Medicine) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Seminar promoted lots of thought and questions - and plans for more work
Year(s) Of Engagement Activity 2022
 
Description Talk at BBSRC meeting in Beijing 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Much better cohesion within all the Newton funded projects
Year(s) Of Engagement Activity 2018
 
Description Talk to Bogazi├ži University Science Club (Turkey) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact Part of their seminar series of scientific enlightenment - with a focus on evolutionary biology.
Year(s) Of Engagement Activity 2022
 
Description University of Coventry Seminar on phages, phage therapy and phages in animals 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Seminar
Year(s) Of Engagement Activity 2022
 
Description Virtual FIS (Federation of Infection Societies) 2021 Conference hosted by the British Infection Association 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact 15 min presentation (pre-recorded on Oct 21) + 30 min Q&A
Part of the session - Future Proofing Antibiotic Resistance: Alternatives to Antibiotic Discovery
Year(s) Of Engagement Activity 2022
URL https://www.britishinfection.org/events/fis-conference