From curiosity to curative- developing Bdellovibrio as living antibiotics between farm and fork

Lead Research Organisation: University of Nottingham
Department Name: School of Life Sciences

Abstract

Bdellovibrio are small, human-friendly, predatory bacteria which invade the cells of other pathogenic bacteria (such as ones that give humans food poisoning) and kill them. Bdellovibrio have no activities against human, animal or bird cells but they are naturally good at killing other bacteria. Pathogenic bacteria do not have simple cell surface receptors for Bdellovibrio attachment and invasion and so it is not easy for them to develop resistance to the Bdellovibrio (unlike the situation with conventional antibiotics) Bdellovibrio were discovered in the 1960s and recently we and others have been researching their genes and trying to understand how they may be useful to humans. Previous scientists have published short reports showing that Bdellovibrio has predatory activity against a range of food-borne pathogens, from farm settings, but they usually studied this on food preparation surfaces such as boards or knives, or in foods themselves. No-one has published data on Bdellovibrio treatment of food borne pathogens by Bdellovibrio in farm animals. This leaves Bdellovibrio sitting on the sidelines with much reported potential as an antibacterial agent but no real steps forward in applying it. We wish to do this in our application with human pathogenic Salmonellas including the phage type 4 Salmonella Enteritidis which causes human disease in Europe UK and USA, and in poultry and with bird pathogenic and colonizing strains including Salmonella gallinarum. We have carried out a small pilot project that shows that giving chicks that are colonized with Salmonella, some lab strain Bdellovibrio to drink, does not harm them, but does reduce the numbers of Salmonellas a lot in their intestines. Now we are asking for the funding to be able to study what the best dose of Bdellovibrio is for getting rid of the maximum amount of Salmonellas. How the Bdellovibrio do it exactly inside the chicks and what factors are important, and whether if we use Bdellovibrios we have isolated from inside chickens poultry farms, these ones are better than the lab Bdellovibrio at removing the Salmonellas without harming the chickens, or the people who may go on to cook and eat them. We will work with a mathematical modeller on all the data we collect to calculate what is going on inside each chicken's gut as the Bdellovibrio are killing the Salmonella and to work out how this might prevent infections in flocks of birds. The idea is to develop Bdellovibrio as a real new treatment for human, animal and bird disease and this is the first step.

Technical Summary

We are applying to use predatory Bdellovibrio bacteria as a living antibiotic against Salmonella in chickens, particularly timely given recent EU Zoonoses Legislation (Directive 2003/99, Regulation 2160/2003) which requires member states to have National Salmonella in poultry control programmes operational by 2008 and by 2009 no eggs from flocks which are infected with S.Enteritidis will be allowed to be sold into the fresh egg market. We carried out a small, pilot study that proves that our concept that orally delivered Bdellovibrio do kill Salmonella colonizing the intestines of chicks; that the Bdellovibrio do survive gut passage well. We wish to extend that study to titer the efficacy of Bdellovibrio in Salmonella treatment regimen and any longer term effect of Bdellovibrio on bird health and also to test the behaviour of Bdellovibrio mutant strains that we have show to have reduced predation in vitro, in the chicks; analysing and modelling the data gathered to determine the important parameters in the Salmonella reduction process and their possible impact on flock level infections . We will also carry out parallel in vitro predation assays to see whether in vitro assays can predict in vivo predatory behaviour. In pilot isolations from 25 different birds on several different farms, we have found that Bdellovibrio and like bacteria are rare constituents of the normal flora of some, but not all chicken caeca. We will test the prey range and predatory efficiency of these Bdellovibrio-chicken isolates in vitro and if strains with enhanced anti- Salmonella properties, over our existing lab strain, are found; we will test these in vivo. It is our aim to develop predatory Bdellovibrio bacteria as true friendly bacteria, that are safe and effective on the farm for the removal of food-borne infections of birds and also later as treatments for other infections of animals birds and man. This application is the first step along that path.
 
Description The key findings of this project were:



1) That predatory Bdellovibrio bacteria can be administered orally to living animals (in this case poultry) without adverse effects on their well being and growth.



2) That if the bird's guts are colonised with Salmonella infection (important as this can spread also to humans) then the Bdellovibrio a) reduce the gut inflammation caused by the Salmonella, b) reduce the Salmonella numbers in the gut by 90% with a single oral dose.



3) The Bdellovibrio were short-lived in the gut (48hours) so repeated dosing would be required for an active therapy, 4) the Bdellovibrio were not shed live into the pens or the drinking water of the birds so no extra spread in a farm setting would be expected (though in our experiments the work was done in contained animal houses, not on farms).
Exploitation Route Animal and human health. Under investigation. We are exploring exploitation follow ups currently.
Sectors Agriculture, Food and Drink

 
Description Used in a further project investigating the safety of Bdellovibrio as a therapeutic in other living creatures. US Govt project. This led to another paper (Willis et al Current Biology 2016) showing that Bdellovibrio can be used as live injected therapies. Without this first BBSRC project we might not have gone on to that next work which has been well received. The original Atterbury Hobley et al paper (2011) from the BBSRC project has been cited quite a bit recently by groups thinking about live bacterial therapies.
First Year Of Impact 2015
Sector Government, Democracy and Justice,Security and Diplomacy
Impact Types Policy & public services

 
Description Co-authored a blog with Carolyn Shaw of PEW Trust on Targeted Research Approaches to Antibiotic Alternatives
Geographic Reach Multiple continents/international 
Policy Influence Type Participation in a advisory committee
URL http://www.pewtrusts.org/en/research-and-analysis/analysis/2017/01/05/out-of-the-box-approaches-can-...
 
Description Contributed to PEW Trust Consultation on New Scientific Approaches To Improve the Antibiotic Pipeline
Geographic Reach North America 
Policy Influence Type Participation in a advisory committee
URL http://www.pewtrusts.org/~/media/assets/2016/05/ascientificroadmapforantibioticdiscovery.pdf
 
Description DARPA Pathogen Predators
Amount $7,000,000 (USD)
Organisation Defense Advanced Research Projects Agency (DARPA) 
Sector Public
Country United States
Start 04/2015 
End 03/2017
 
Description Leverhulme Trust Research leave Fellowship
Amount £45,000 (GBP)
Funding ID RF-2013-348 
Organisation The Leverhulme Trust 
Sector Academic/University
Country United Kingdom
Start 10/2013 
End 09/2014
 
Description A panel Q and A membership at a University Chancellors Lecture event in Royal College of Physicians with Lord O'Neill on Antimicrobial resistance 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact I was one of three panel members answering questions on new approaches to combat AMR after a short presentation by Lord Jim O'Neill to an audience in London.
Year(s) Of Engagement Activity 2017
 
Description Gave Aberdeen Annual Microbiology Lecture Sept 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Many questions from schools audience

None but received appreciative feedback
Year(s) Of Engagement Activity 2013
 
Description Gave an invited Darwin Lecture to the Secular Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other audiences
Results and Impact Gave the invited Darwin Lecture to the Secular Society explaining how an evolutionary genetics lab thrives best staffed by people of all faiths and none and how sampling bacteria world wide, including with the help of religious sites where waters may have a particular characteristic can lead to new science discoveries. The idea was to show that faith and science are compatible and to have a healthy debate. Which we did.
Year(s) Of Engagement Activity 2018
 
Description Hosted school student for 1 week research in lab 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Lower 6th pupil experienced hands on Biology research prior to deciding on University applications.

Student applied to University to do Biology
Year(s) Of Engagement Activity 2014
 
Description Interview on BBC Radio 4 Inside Science 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Interview on recent paper using injected predatory bacteria as cures for infections in zebrafish.
Most of this work was funded by DARPA but some participation and relevance of BBSRC projects
Year(s) Of Engagement Activity 2016
URL http://www.bbc.co.uk/programmes/b082vz39
 
Description Interview on BBC Radio 4 The Life Scientific 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact I was interviewed in an episode of Life Scientific on BBC Radio 4 discussing my labs work on Bdellovibrio in its entirety. We have since had many contacts from a company, from other scientists and members of the public in UK and abroad where the podcast was available
Year(s) Of Engagement Activity 2017
URL http://www.bbc.co.uk/programmes/b08mb1fm/clips
 
Description Pre Pint of Science Talk on Predatory Bacteria at Canal House Nottingham 10th Feb 2017 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Gave a public talk in a Pub 7-8pm at the Canal House 48-52 Canal St, Nottingham NG1 7EH for Pre Pint of Science. Topic was predatory bacteria as living antibiotics
Year(s) Of Engagement Activity 2017
 
Description School visit 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact 14 year old school pupils (60) visited labs and interviewed our research group

Very positive feedback...school is in disadvantaged area..we tried to raise their aspirations re higher study
Year(s) Of Engagement Activity 2012
 
Description Took part in a Pew Foundation Panel and meeting on the Antibiotic Pipeline and alternatives to antibiotics in Boston USA 
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 Took part in Pew Trust Panel answering questions on alternatives to antibiotics from physicians and pharmacists policy makers and scientists. also contributed to part of the Pew Antibiotic pipeline report- named and thanked for this and a co-authored a blog with PEW
Year(s) Of Engagement Activity 2016
URL http://www.pewtrusts.org/en/research-and-analysis/reports/2016/05/a-scientific-roadmap-for-antibioti...
 
Description Was on SfAM panel on antimicrobial resistance at a meeting in London Royal Institution of Civil Engineers 24th November 2016 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Sat on a panel and took questions from the audience on antimicrobial resistance and how to tackle it.
Year(s) Of Engagement Activity 2016