[Music] thank you very much for the invitation to talk today and thank you to the chair for the introduction so yes compiler boxer is the harmless comment sort of broiler chickens how many times have we seen that in a paper hundreds it's still even today is still quoted that it is merely a harmless commensal of compiler batter and if I can do anything in the next 35 40 minutes is to convince you they're actually compiled about is not the harmless commensal that we think it is okay so compiled of actor as a pathogen in the UK it is really really important to study in its own right it's the leading cause of foodborne zoonosis worldwide and actually compiled about sir is a good model to study environmental and management influences within a poultry house and also within animal production as well there are a hundred and ninety thousand cases across the EU so this isn't a small pathogen that we're talking about those are confirmed cases so for every one of those confirmed cases it's been estimated that there are further nine in the community this has a massive draw on the healthcare resources it's estimated that there's approximately three cases every two minutes so if we were to work that out for the duration that we're spending sat in talks today that would be quite a lot of people affects one percent of the EU population so again not not small and not insignificant so this is data from essa which is from 2016 and you can see the big orange bar at the top that's compiler beta that's why it's so important that's why we invest heavily to study it is closely is well I say closely followed actually it's not closely followed but Salmonella is its next biggest rival and in comparison to some of the other diseases that we focus on its way I in the lead ok so where is it where is it from well actually find in most animals so it's finding sheep pigs cows wild birds companion animals it's also find in water it's find in the environment it survives quite well in the soil it's finding unpasteurized milk pets with diarrhea are quite a big reservoir but actually the biggest culprit is chickens so chickens in compiler box a source of go hand in hand so here it is this is what I have spent the majority of my career to date studying I first came across compiled about her in 2004 so we've had quite a long friendship it's had its ups and it's downs just when I think I understand what it's doing and I think I've got the upper hand it throws me a curveball and I have to go back to the drawing board and think again but yeah here it is a tiny little spiral shaped bacterium with Bipolar flagella okay so just so that we're all sure what compiler bacio is going to cause us if we are unfortunate to get it well the classic symptoms of other foodborne pathogens vomiting diarrhea abdominal pain fever chills if you're really lucky you'll get a mild infection so you know a couple of days feeling under the weather and and you're okay actually in some cases it can be very serious it can last from a few hours to several days most infections are self-limiting so we don't need antibiotics to treat them but in some cases particularly in immunocompromised actually it does cause a bit more of a problem it's not trivial w-h-o is name compiled of actress the number one foodborne pathogen it's thought not to grow in food but survive in food and it has a low infectious dose of less than 100 cells okay so we used to think compiled of actor we'll just kill it by cooking we'll kill it by processing in the poultry pan there's no way it's going to survive a skull tank everything we know about compiler Vactor tells us that it's a weakling and unless the conditions are absolutely ideal it likes 37 degrees it likes 41 point five degrees the body temperature of the chicken needs a micro aerobic atmosphere there's absolutely no way that it will survive a skull tank it does it'll quite like skull tank you can isolate compiler back to quite well from some skull tanked Walter it's actually a lot more heat resistant than we thought and it can survive throughout the whole of the poultry processing chain it's got a long incubation period so you know we could have all eaten some chicken for our lunch today which I'm sure would be fine but you know if in 3 to 10 days you're starting to feel under the weather then just think back if there was chicken today and that you ate it people always tend to blame the takeaway dodgy takeaway that I had the other night that's what caused my foodborne illness actually the majority of people actually poison themselves so we have consumers that are handling chicken in the home some of these carcasses have up to 10 to the 9 bacteria on them and they spread it nicely around their kitchen cross-contamination is quite profound in that case and they poison themselves but of course it's never that it's always that take away a few days ago so yes you can get high temperature with it you can get abdominal pain diarrhea which is often bloody particularly in children and one percent of cases have long-term side effects so if you're really lucky and you get a particular serotype of compiler boxer it's gonna stay with you for the rest of your life so it'll be your your friend probably not your best friend it could give you reactive arthritis it could give you autoimmune diseases if my boss was stood on the stage today professor Tom Humphrey he would tell you that his daughter had compiled about her when she was a teenager she was one of the unlucky ones and is now in a wheelchair and suffers from reactive arthritis along with autoimmune diseases elderly people are more at risk immunocompromised are also at risk most of the deaths occur in people that already have conditions such as bowel cancer so compiler actors not a killer in the majority of cases it only kills a few people so I think that makes it okay right it's you know not headline news all the time so it's fine but chicken is the most important vehicle and it is blend for the majority of cases we can link back with traceability we can link back chicken as being the source in the majority of human cases it doesn't matter where in the EU you are your chicken flocks will have compiled of actor so this is quite old Dayton I this is from 2008 but this is the number of positive boiler batches in the EU so in the UK at the time we were looking at seventy five point three percent of our boiler batches that would have compiled about sir Spain and Portugal was slightly higher in in the 80s I guess the best place to be would be Denmark who had at a time nineteen percent so it is unless you went further I to Finland Finland are really good three point nine percent back in 2008 their numbers are still low probably something to do with the climate so they have quite harsh winters I'm in Finland so they have lots of negative boiler batches during the winter months okay so I said that it doesn't make headlines actually sometimes it does say not people die in but actually the headline here in the Daily Mail deadly chicken cover-up so yes our supermarkets in the UK have it says six in ten supermarket birds have lethal bug but watchdog watchdog won't say what stores they are so the misconception was you know that if you were buying a chicken that was organic you would paid quite a lot of money for it from one of the more upmarket retailers it wouldn't have compiled about two actually that's not true so and they now do name in shame so that is data from the UK supermarkets that will show which boilers are positive in hi many of it big incentives within the UK to reduce compiler bacter so quite heavily led by the retailers after headlines like this and work is still ongoing in that respect okay so is it a commensal is it a pathogen well it's got a long history of being called a commensal it was thought not to impact on Birdwell fare or bird health it's in the majority of birds at high numbers doesn't mind what bird it's in an organic and free-range a barn reared there's no visible signs of disease so that makes it a commensal right can't see an affair no clinical signs it must be it must be a commensal so for years and years and years that's what we thought it was a harmless commensal in every single chicken flock just sat there not doing anything but actually it's quite a lot of evidence now to suggest that it's not the harmless commensal that it's thought it was thought of so chickens have changed a lot in the last 60 years so what we've got is a chicken am from 1950 at day 68 in the top image versus a chicken at day 47 from 2008 so we've changed our chickens we've pushed them to give us more muscle because that's what we want so in doing that we have changed the chicken dynamic so we've bred them for to be heavier birds to be more efficient at converting their feed so actually this has had an impact on compiler bacter in itself so the bottom image is 1957 versus 2001 chickens and think it's quite profound there that you can see that the 2001 chickens have a lot more muscle we need to do that because chicken is in demand lots of people eat chicken it's everyday meat that we eat is not like it used to be where it was eaten on a special occasion so it's a big business this is not small and it's estimated that global chicken production will need to quadruple to satisfy demand in the future so this isn't going to go away this is only going to get worse and actually by doing that with our chickens what impact are we then having on the compiler boxer so if chicken production is going to quadruple compiler batch is not going to go anywhere so there's two health threats from compiler boxer and chicken so there's high level surface contamination on chicken carcasses up to ten to the nine CFU per gram has been estimated so this is a cross contamination risk with consumers in the kitchen but actually it's also a cross contamination risk during slaughter and that's where a lot of the surface contamination comes from so we have negative broiler batches that can go into processing as negative they come out the other side as positive on the surface so that's historically where a lot of the controls were aimed at it was aimed at to reduce surface contamination in processing because that's where we thought the biggest problem where so we did lots of interventions in the poultry processing plants in able to limit surface contamination to try and control compiler batter so we thought that that was quite good and we were sort of winning the battle and then compiler batter is telling us that actually surface contamination is it's not we're not we're still seeing a lot of cases so actually maybe there's more of a problem so that's when you know you start looking in a bit more depth and you're seeing contamination of liver and muscle tissues so this clear increases the risk when you're under cooking it because it's there in our meats that were eaten but actually where is this contamination coming from well actually if it's in the deep muscle and it's in the liver it's not gonna come in from cross-contamination during processing there's good enough controls within process in knife so actually what's happening is compiler butter is leaving the gut chore in ririn so it's leaving the gut it's going out there is into the bloodstream is it's infecting the liver is happily sitting in the chicken muscle which is increasing the risk to the consumer so chicken liver pate people like this is it's had a bit of a revival recently with lots and lots more people eating it is a dinner party favorite and we're now seeing a lot more outbreaks in humans being able to be linked to chicken liver pate and damage to the growler gut say whether that's damaged by inflammation from another bacteria whether that's damaged by other other pathogens or by worms etc helps compiler back to to spread to the tissues so compiler actor doesn't do everything on its own it makes friends with other bacteria in the gut and to do it it's quite good friends with e.coli they work quite nicely together too and break down the tight junctions to help it spread so actually intestinal spread and chicken livers well these are just three screenshots of outbreaks in the UK that have been linked to chicken liver pate and eating pink chicken livers this problem isn't going to go away it's only going to get worse our eating habits change our food habits change there's a trend to eat meat slightly raw or rare and that trend is is happening in chicken so there are some people who prefer to eat slightly raw chicken I think they like to live in on the edge for that I definitely wouldn't be touching undercooked chicken the risk is too high but there is a food trend to be able to do that so we've got some evidence which shows that in some chickens see to June I know that m1 is a particular strain of compiler back to GG night it was isolated from a human and a poultry flock at the same time the human had been in with the poultry flock and this Chasen damage to the gut epithelium so what we've got here is we've got uninfected birds and we've got to breed different breeds of birds one that grows quite fast and one that grows a little bit slower and what it what it's showing is that in the uninfected birds we've got lovely nice healthy-looking guts actually when we put compiled about 2m one in the faster-growing birds we're seeing a shortening of the villi was the infusion of the villi we're starting to see some inflammation and the effect wasn't as great in the slower growing birds but there is still some changes to the gut structure so damage to the gut and diarrhea associated with the dysregulation of inflammatory responses can happen with seated unites m1 so this study was published in 2014 and at the time it was when Professor Humphreys Greek were at the University of Liverpool and they thought that this was quite this was quite unusual because what they actually saw was that seated unites in these birds was causing diarrhea they were quite excited about this because it was probably one of the first recent evidence that but chickens given seeded you and I am one could get diarrhea at the same time I was still working at the University of Bristol and I was writing a book chapter on whether or not compiled about sir was a commensal or a pathogen in poultry be in the conscientious postdoc that I was at the time and still AM I went back to the literature and I find a study in 1984 where they took some chickens 288 chickens and they fed them with live cells of seeded unites isolated from diarrheal patients okay say what this shows is that there was evidence back in 1984 that if you give chicken a strain which has come from humans with diarrhea it causes a problem and causes diarrhea within the chickens okay so at the time I wasn't very popular with with Tom because I had just basically crushed his theory that he was going to be one of the first ones which show that compiler back to cause his diarrhea in chickens it hasn't affected it he did offer me a job in Swansea four years ago so it's okay we're friends again now so this study in 1984 diarrhea in chickens okay this study isn't heavily cited by the compiler back to research community it was published in quite an obscure journal which might explain one of the reasons but actually at the time it just didn't fit with the dogma of compiler backs have being a harmless commensal if it was a harness commensal it wouldn't be causing diarrhea back in the 1980s in chickens so this study went on to have more images where they showed distended intestines in chickens given C to June I so again evidence back in the 1980s that perhaps compiled about it isn't the harmless commensal there was being referred to at the time okay so this study went on a little bit further and they actually looked at whether there was infection of tissues and no surprises there was so they looked at the spleen the liver and the and some blood and they actually found compiled about in these chickens but 48 hours to 168 hours after feeding okay so this is evidence you know the systemic invasion was happening right back in 1984 so compiler bacter can and does have an impact on the health and welfare of chickens have a direct impact on broiler performance now of course the caveat that I always say to this is it does depend on which string of compiled of actor and the bird type just like we are all different the chickens are also different so no two birds are the same no two compiler batters are the same so compiler batter does have an effect on the feed conversion ratio compiler backs or infected birds tend to have to eat more to maintain what to gain and maintain their weight but they're less efficient at converting this food if they're eating more then the consequence of that is that they're going to the toilet more and that leads to wetter litter wetter litter leads to four and leg problems and we can see in experimental studies where we have an increase in hock marks and polo - Emma Titus in birds that are infected with compiler boxer compared to our uninfected groups so again the impacts on weight gain and health isn't new so this is a paper from 1981 which actually shows a difference in the body weight between control and experimental birds and experimental birds in this study our birds that were given CGG night and it also shows that there is an effect on mortality and an effect on diarrhea so what we thought we were studying it was new phenomenons actually had already been published over 30 years ago and again this study has been largely overlooked so bring enough a little bit back up to date with some more recent findings so this is some data from some studies metric erudite in Scotland and in Ireland and these show that compiler back to negative farms at fin so in the UK we still follow the practice of thinning our flocks so this is where we overstock and a few days before slaughter about five days before they reach the right age we remove a proportion of those birds so that they can carry on for five or six more days and so there's still at stocking density at their life and so when we do Finan obviously we break down biosecurity because we're taking in equipment and people and so we do tend to see an increase in compiler actor in our flocks between Finn and between clearance but this is this is telling us that compiled about to negative farms that finn had better feed conversion ratios than farms that were positive at the thin time point so this is showing us that actually compiled about to does have an impact on feed conversion ratio and so actually then gives us a little bit more of a tool and a bit of leverage that we can use to say that actually there's an economic impact here of having compiled about sir within the flock so there's quite an interaction between chickens and compiler bacter so we could regard it as an opportunistic pathogen in broilers maybe like e.coli or clostridia but actually we could consider it to be a true pathogen in chickens it is causing them disease it is causing them harm and it is causing an impact on their health and welfare so of course the bacteria may more easily infect animals that are compromised by poor health and poor welfare so if we do everything else right we've got a chance of controlling compiler bacter if we've got a lot of unhealthy birds in the in flocks then actually compiler back to takes advantage of this and it spreads a lot quicker and it establishes itself a lot quicker so we've got some studies which have shown links between wetter litter and actually e-coli so if we've got you coli infected birds compiler boxer seems to proliferate and compile a batter that is excreted by the first infected bird means that it actually then more easily infect others and it spreads rapidly throughout the flock so the frequent culling of these sick birds may reduce the risk of koala bacterial infection how we identify which bird is sick well that's heavily reliant on farmers knowledge of poultry but it could help to control compiler batter within the flocks got health is really really important so we've got some key questions here which is does inflammation play a role in the extra intestinal spread of compiler batter from the gut if it does would controlling inflammation help how are we going to control inflammation in the gut well actually we can give feed additives which could improve gut health there's been lots of previous studies on see people have looked at prebiotics probiotics essential oils medium chain fatty acids singly in combination one study through everything that was known to have an impact on compiler bacter or improved gut health into the chicken it didn't work so if you throw everything out the chicken a compiled about to just kind of quite likes it and takes advantage of it so what we did was something slightly different so we looked at supplementing diets with short chain fatty acids so we did this youth in salmon oil so salmon oil is high in polyunsaturated fatty acids it is a byproduct of the salmon oil the salmon farm in industry they use a little bit of the salmon oil at the time the rest of it was was literally thrown away and we've got some evidence which shows that a number of diseases in Western societies are thought to be due to an imbalance in polyunsaturated fatty acids and we know from history that actually chickens way back when in the wild would have consumed leafy plants and actually they would have had far more short chain and three fatty acids than their domestic grain fed counterparts that we have today and some studies at the University of Bristol on laying hens showed that dietary shortchanged c18 and Fries decreased fractures and enhanced bone strength in laying hens so there's some evidence there which shows that in feed in fatty acids does have an impact on the chickens and n3 polyunsaturated fatty acids can also increase intestinal barrier function and have anti-inflammatory effects so actually the signs are looking quite good here that this is probably a good target to go for so reducing inflammation could control compiled about sir we know that compiled about set acts with interferon gamma to break down tight choose between cells this causes a feedback loop of inflammation alliant allowing high numbers of compiler boxer across the epithelium but there's very little work on this area and none that at the time have used n3 polyunsaturated fatty acid diets to control compiler boxer infection so our aim was really a simple one would feed in chickens and n3 supplemented diet control compiler back to colonization so we took Ross broiler chickens we stopped them at commercial stocking density we reared them on standard diets that were supplemented with either nun 0.5 0.75 or 1% salmon oil with the levels of salmon oil being reduced by half in the finisher and withdrawal feed we infected them was compiled about 2 to Jun im1 when they were 21 days old this mimics the two to three-week infection first site that we see commercially we took ten Birds at 714 and 21 days post infection we looked at the liver and we looked at the cecum we scored them for hawk marks and powdered dermatitis we obviously recorded mortality throughout the study and we were interested in their weights so what happened well compiled about two numbers in the cecum at 714 and 21 days post infection so what you see here by the asterisks is where there's a significant difference so it's 7 days post infection we saw a significant reduction between the control and 0.75 percent supplementation by the time we got to 14 days post infection we saw a significant difference between the 0.5% supplemented and 0.75 present supplemented so this showed 14 days post infection it diets containing salmon oil could reduce compiled of actor by between 90 and 99 percent so this insignificant we're seeing up to a two log reduction in Kampala bouts of colonization now I've put the 21-day post-infection graph up there to show you two because it fits with my theory that not all chickens are the same because obviously the birds that we examine seven days post-infection well those are gone they're dead these birds that we looked at at 21 days post infection there's no significant difference here between supplementation so is it that you know we've we the supplementation has done its thing and now he's actually predisposing them to disease or is it that we because we've lowered the level by half is it then it's just not having an effect so those are still slightly unanswered questions if we look to the livers at seven days post infection and we saw significantly reduced livers in the compared to the control in the point seven five and one percent supplemented and also the point seven five to the zero point five and the one percent to the zero point five supplementation by fourteen days post infection and supplementation was still having an effect where we saw reduced livers in supplemented diets and again by 21 days post infection we didn't really see any positive livers in any of our birds one positive liver in our own supplemented group so the number of positive livers did decrease with increase in polyunsaturated fatty acid in the dire mortality so yeah there there was an effect while mortality so we saw less deaths in our supplemented diets interestingly in the one percent supplementation we saw higher numbers compared to the other two supplementation so actually in this case are we taking it too far so is one percent too much and we're actually predisposing them to diseases and Hawk marks on this graph are in black pea dermatitis in gray this is at 21 days post-infection and we can see that supplementation did reduce the number of hop marks and powder dermatitis that we saw so compiled about two numbers were reduce significantly in the seecombe by some of the n3 polyunsaturated fatty acid diets as was the frequency of Kampala Boxer positive livers the number of Kampala Boxer finding the livers did not differ significantly between the diets for those sample sizes were quite small we did only do 10 Birds for each one of these time points so these data demonstrate that the addition of salmon oil to the diet appears to reduce compiler back to numbers in the intestine and the number of birds carrier in the liver and I'm sure the question on your lips is that you fed chickens salmon oil therefore do chickens taste of fish and I can categorically say that the chickens did not have any fishy taint the reason I know that is because we reared some and we sent them off to be eaten by the University of Bristol taste panel basically they're trained eaters and there was no fishy tank in the meat at all so well what we need to say is that actually this needs to gain a large scale so this was quite small groups of birds we also need to understand a bit more why so what effect is this having and also what effect on the microbiota this also needs investigating so we wanted to in another study assess the impact of poor welfare so this time we used large industrial scale animal experiments we had over a thousand birds per group we had stressed Birds which we it either injected with six hydroxy dopamine or they had gone through production stresses so this project pet name was life end I am assured it was the life end of the chicken and not my life end as the postdoc working on it at times it did feel like my life was ending we had time points at key events during the last 24 hours of the birds life that meant I got no sleep for 37 hours the 6 a.m. point was really funny for some reason I don't know why we took the seeker we took the ileum and we took the liver we also looked at the secret on sauce so that we could do immune responses and we examine the microbiota so what we can see here is this is just show in a few of our time points so I'm showing you pre-stress so this is before any stressors had happened catch in mid transport and it final kill we did two methods of stunnin in our harvest group it final kill we use gas stun in and we use electric stun in there was no difference between the two stun methods and what this is showing is actually that there are some key points within the production and the life end of the chicken where a compiler back to levels fluctuate we took these secret on cells at the first and last time point and we measured the immune responses using real-time PCR we examined are four or five or six hour late i'll 1001 beta interferon gamma TNF alpha I was 17 and I'll 18 I'm only going to show you a selection you'll be pleased to hear so what we've got here is our six i'll eight i or 10 and i'll one beta and what we're seeing is that there's a compiler back to effect because we've got a difference between our untreated birds and our control birds and actually in some cases there's also a stress effect so there's an effect of giving these birds six hydroxy dopamine six hydroxy dopamine mimics the stress response that chickens would have it's quite an acute stress but actually it shows an impact on the immune responses as does certain stages of the life and production cycle so what this project um showed us actually was their stress is not is important during the last stage of rearing as previously thought so what it was assumed is that stress birds at their life end would have higher compiler back to levels and that's why we were seeing numbers in the processing plant because these birds were stressed and what this this animal experiment has shown is that actually that's not the case now stress earlier on in the chicken's life could actually be really important because we saw in a stress bird that counts in the seeker were higher than counts in the ileum and actually it could just be this movement in the gun stress with either the harvest or 6 hydroxy dopamine did have an effect on the immune response with greater responses compared to the control in the stress groups and Kampala bats are had an effect on the immune response I would love to be able to show you the microbiota data and that's still being analyzed it's been done as I speak to you now and the sequences are all back and we're just working our way through them so not all compiler boxes are the same so this is a hundred compiler boxes from different sources and so we've got human strange chicken strains wild bird cattle strains and this is them being infected into two cell lines so HT 29 s a human cell line and a e 11 clone of a chicken cell line and what you've got here is are late responses and you can see that there is a diverse response to the compiler batches so not all strings are the same and this is some new data from us which we will be sending off for publication and shortly so this was looking at an organic free-range flock in the southwest area of the UK we took the seeker the ileum and the liver from these birds and we cultured them and then we put them back through our a 11 cell line and what this shows is that actually isalus that have been involved in extra intestinal spread are more invasive in vitro than isolates that came from the liam or the cecum so what are the key messages well I hope that I've convinced you that compiler banter is not a harmless commensal we've got data from the field and the lab for over 30 years which shows that this isn't the case is it cannot be collapsed as a harmless commensal it affects bird health their welfare and their performance an impact does depend though on the bacterial strain and the bird type we've got a real change in host pathogen dynamic so chickens are very different from what they were and compiler boxer is very different from what we thought it was it can mutate and it can take up DNA from other bacteria it doesn't like to do all of the work itself exposure is difficult to prevent and perhaps the best way forward is to improve resistance of chickens to compile of actor so I would just like to acknowledge a few people so a lot of there all of the salmon oil where it was done when I was still at the University of Bristol and I couldn't have done it without the help of the three people there up on the screen there under the University of Bristol logo I'd like to thank the life end project consortium so that's Nick's box SRU see Stephen rushed on it Newcastle and Julian Kelly at Leicester and BBSRC who funded that work thank you very much [Applause] [Music]