[Music] okay well I guess I sort of wanted to start my presentation talking a little bit about antibiotic use in the United States and how that has changed recently Todd Todd introduced the idea that through nae we have started to reduce the use of antibiotics within the livestock industry and our FDA has actually come out and is providing some information now on there they're talking about it as usage but really if you look at it this is sales and distribution so these aren't a direct one-to-one relationship between a sale and a utilization in animals but they do give us some sort of indication of what's going on from a livestock species we have I think they have they've released information going back to 2009 but I've just summarized 16 to 17 here and what you can see here well probably the first thing that jumps out to you is that at least in the US cattle and swine are using the majority of the antibiotics followed by turkeys chickens and then other and this is the FDA is actually breaking this down into medically important and then also non medically important so this is the the medically important so what you can see here is from a poultry standpoint I think all of our species are generally reducing their antibiotic use but from a chicken standpoint you can see a fairly large reduction here almost a halving of the medically important antibiotics that we're using currently now if we look at the non medically important it actually tells a little bit of a different story here and you can see the non medically important end for poultry and Cattle these are almost exclusively 90% plus ionophores so we are still having some challenges and and we are still using some ionophores out there what what I'm going to predict is that when FDA actually releases this information for 2018 as we've really ramped up that na production at least in the United States this number here will continue to go down for chickens but overall I just wanted to provide a short update on sort of where the u.s. is in terms of antibiotic use at least according to the FDA and I wanted to use that as an introduction to this talk so that I can talk about okay well what are some of the consequences that we've run into from that that standpoint so so really I'm gonna start talking about coccidia infections first and then I'd like to talk a little bit about Clostridium infections as well you know we usually say coccidia and we talk about that as we have coccidia but as everyone knows there are several different species that have that affect chickens different species that affect turkeys as well and they don't infect all the same place so I think this is something important that we need to we need to consider as well is a serval ina is actually going to be infective in the upper GI tract the duodenum and and that upper area and this is where when we start talking about a coccygeal infection we need to be a little bit specific because if you have this infection in the duodenum what you're going to see is you're going to see a larger effect on energy utilization I'll talk about that a little bit later but some of our other Ameria species can affect the our infective elsewhere this is actually Tinelli in the Sica and here's where we'd see some different signs here you'd actually see some some blood in the in the excreta but I wanted to talk a little bit about this is this is a little bit dated now but this was the most recent estimate that I could come up with 90 million to control in the US and 300 million in losses I think especially transitioning to the removal of ionophores that's going to go up tremendously but I actually haven't seen any data released for that so I just wanted to go back and talk a little bit about really what are we looking at for for coccygeal infection here is a bird that is really really not doing well this is a very very serious very serious coccygeal infection here you can see this bird you can see there's some stunted growth there you can see that the head they're still covered and down you can see though this bird does have substantial feather cover but those feathers are ruffled up certainly not doing well you can see discolored stool here and it's a little bit hard to see projected up here but you can also see a little bit of lightning here in the in the shank area especially lightening up here we compare that to a similar age broiler you can see the vast differences between these two birds so with that just some causes of this certainly this is ubiquitous to chickens and turkeys or turkeys We certainly have some seasonal effects here and a lot of this goes back to this litter moisture because that's important for the sporulation process but we tend to see this more in the winter and fall and that sort of makes sense as temperatures get colder we're not moving as much air through our houses and we're going to end up with a higher moisture content looking at that spore ulation and that's really where we're gonna see most of our problems the other thing that we're dealing with is is we're recycling oases every 6 to 7 days so you might have a a minor challenge to begin with but that's going to build over time hopefully that those birds will also build immunity over time and that's sort of what the coxy vaccine is there to help provide but what we see is when it gets real bad it's usually after probably 3 maybe 4 cycles and then we get we get a serious outbreak so I sort of want to transition here and talk about start talking about necrotic enteritis as well but there's actually very good link between the two and I'm sure at this point I'm not saying anything that that's not not familiar to you guys but one of the one of the one of the real causative agents and in most of the models that we use for necrotic enter itis we actually have some sort of coxy challenge and there's a couple reasons for that one when you get the coxy challenge you get the the coccygeal spores are going to enter and actually infect cells and that does a couple of things one it's going to decrease that gut integrity so you're going to get leakage of plasma proteins into this intestinal barrier area you're also the body is going to respond to that and it's actually going to increase mew copra mucus enos's so it's going to increase the amount of mucin in that area so why is that important when we start thinking about Clostridium perfringens really it needs a couple things to really bloom one is excess protein which is going to come from these plasma proteins leaking into this area the other is mucus it's actually thrives and does very well with excess mucus so what we're seeing here then is after we have a coccygeal infection we really open up some sort of challenge some sort of insult that actually allows the Clostridium perfringens that is there naturally to bloom and become more of a challenge for us so if we look at this from a from a growth standpoint these are just some examples you can see the lesions are very much different but you can see still there's a lot of damage here these are generally in the in the small intestine area you can see in very serious cases there's a lot of disruption a lot of damage here there's a little bit newer estimate here and it's estimated with one to 1.2 billion in losses the other thing that I want to point out is as with some of the coccygeal species when you have a necrotic outbreak you can definitely see this in the in the excreta and I want to come back to that later because I think we've got some issues here We certainly have some issues here with digestion if you just look at the damage caused here and and what's coming out the back end that certainly changed that digestive capability of that bird so I do want to get into a little bit more about the the metabolism and how that hosts how that bird is actually viewing this challenge because I think it's important to come back and see we know we have this infection we know it's going to decrease production but what is that bird actually doing metabolically to deal with that or to not deal with that so this is just a table and a figure out of freitas 2008 where they looked at plasma glucose in both a the group a was challenged with with a tuxedo challenge so this was just straight Cox you there was no Clostridium put in there at all and then the solid line here are the control birds now what you'll notice first is that the data do jump around but the way they have it presented here these are actually pretty tight so you're gonna see about a 250 milligrams per deciliter concentration of glucose in the blood very very normal for poultry and then if you would order to average these across you're probably going to see about a 15 milligram drop with with the coccidiosis and that I think can be attributed just to that damage that the coccidia are are impairing on the on the intestinal system so you can't pull as much as much carbohydrate and well as much glucose across that barrier system so you see just a slight depression it is a slight depression but it but it is there but I want to talk about lipids next because this is someplace especially when we have infection with a serval ina which is up in that upper digestive tract this is where we can run into some problems with with fat digestion so if you see here this is total liver lipid as a percentage and you see again same same thing that the dark is the control and then this would be the coxy infected birds you can see here starting a day after and within two days we see a significant drop in liver lipid level most of these well the lipids here going to be coming from two sources one from the diet and two from lipid epigenesis they're in the liver so part of this is probably attributed to lack of fat digestion the other part would be attributed due to lower energy status overall but where it starts to get a little bit more interesting in this story is when we start looking at plasma levels so here we have plasma fatty acid concentrations and triglycerides over here and what you can see here even though the liver has returned itself to a more normal metabolic standpoint within about two days what we see in the blood is we see a continued drop in both fatty acids and also triglycerides going on eight days up to almost two weeks so serum fatty acid triglyceride levels require a lot longer to recover here and this gets into some of the ideas that we have around that energy metabolism that Todd was talking about earlier and those huge drops in energy digestibility that we see with coccygeal infection so we've got some digestibility data here and this was done back in dark dr. Parsons lab but we did both some amino acid digestibility and then we also did AME N and what I want to point out here is you'll see there's a little bit of variation here threonine was about 30% 20 percent the rest of them were pretty close to between 12 and 18 going back to what Todd was saying earlier it's not surprising to me that threonine digestibility is going to go down that probably has less to do with actual digestibility and more to do with increased mucin being secreted during this challenge and these numbers are actually generated about four days after infections so going really four to five days so going really into that peak first infection and we actually have data going out after that where we see after this first period of infection the bird actually recovers from a digestibility standpoint fairly well but the thing that I want to point out is here you know if we discard disregard the threonine we see between twelve and thirteen say for an average say sixteen percent reduction and amino acid digestibility here but again from an energy standpoint we see a 30% reduction this is going to go back to that disruption in the duodenum and really a disruption of fat metabolism that we can see carried out through that entire bird starting in the liver and going into the plasma as well so one of the things that we've done a little bit more recently as we started looking at body composition in comparison to some of our different disease statuses and birds and body composition just takes that next step from digestibility and says really how is that bird then able to utilize in or store the nutrients that it's taking in and this was an experiment we did at Virginia Tech I was done in collaboration with dr. de Lewis lab and what we saw is after we challenged these birds we saw about a 7% reduction in lean tissue we have a DEXA there at Virginia Tech and after the birds have been euthanized wildy feather them and take them through that DEXA process but the really interesting thing that goes hand-in-hand with our digestibility is that when we looked at the fat stores in these birds we saw fatty tissue actually had a 17% reduction there so this goes further along with our story that from a coccygeal standpoint we're really affecting energy metabolism fat metabolism specifically and we are really altering not just the body weight of these birds but we're altering the composition of these birds they just it really comes back to an energy deficiency in this standpoint and this is really what's limiting that growth when we get into a coccidia infection so just to summarize sort of my thoughts on the coccygeal infection it's caused by a protozoan marry of species and it's made with multiple points within the intestinal tract so again there's some specificity here it can cause that damage needed to facilitate necrotic enteritis it's not the only thing associated with necrotic enteritis but it is one of the predisposing factors and it reduces performance overall invasion of the intestine causes that immune response that that 10% number that dr. clays has come out with I think is still very appropriate for here it decreases both amino acid and energy digestibility and we certainly see that it changes fatty acid metabolism I think that's where when we see these differences in fat response or in energy response are coming through that lipid metabolism area so just to to continue from a body composition standpoint reduced fat digestion reflected in the larger proportional reduction in body fat compared to that lean tissue so I want to talk a little bit now about necrotic enteritis or the disease the disease associated with unchecked Clostridium perfringens and here actually I specifically will say this about the digestibility values but here in general I think we have a little bit less knowledge about how the bird itself responds to this I actually want to comb through the literature I couldn't find anything from how necrotic enteritis actually affects either amino acid or energy metabolism I was actually sort of surprised by that but with some of the issues we have with replicating that in the lab I really wasn't all that surprised at the end of the day but some of the information that I was able to find is looking at cellular damage so smart eight amino transferase and alanine aminotransferase are basically just indicators if you see them in the plasma they're indicators of cellular damage so what we're looking at here our Birds that were infected with a necrotic enteritis model and you can see there were significant increases in both of these both of these enzymes in the in the plasma so as you would expect by looking at those first pictures you're getting cellular damage or you're getting release of those contents into the plasma I was able to find some amino acid or I'm sorry some enzyme data that goes hand-in-hand with the with the Clostridium perfringens model and what we see for amylase so general carbohydrate we see so so this white bar is the non challenged control and then we have the necrotic and Rytas model here so we see a significant reduction when we have necrotic enteritis so so this is again that cellular damage within the intestine actually reducing that enzyme activity that we see same thing for for trypsin so this would be an indicator therefore protein we see a significant reduction here again indicating a general response reduction in enzyme the interesting thing here is for lipase now this is in the jejunum or the ileum and i realize that a lot of the lipase is coming from the pancreas or the duodenum up higher in the digestive tract but we actually don't see any significant differences here so this is this is sort of the first indication where we might have some difference is between the host response to the coccidiosis where we saw a definite change in fat metabolism and now this secondary bacterial infection where we see a more consistent effect in both carbohydrate and protein maybe less so here in the lipid enzymes we have some plasma glucose concentrations here actually just the opposite of what happened in in the occido infection here we saw an increase but then when we look at triglycerides in the blood we see no change across two different experiments so again some differences when we're looking at blood concentrations of some of our metabolites during these infection models we've got a couple different things here we've got some hepatic fatty acid synthesis going on here these data are sort of contrary in the first model you see an increase in both ACC and FAS this is mRNA expression but in the second data there's really no difference here between the the non-infected controls and the necrotic enteritis model so maybe some differences maybe some up regulation from an mRNA level but no real strong consistent responses here from from a metabolic standpoint I don't want to spend a whole lot of time on these first three bars I'll just make the statement here that Clostridium can't alter fat metabolism but what I do want to focus on here is this last column this abdominal fat so this is just an abdominal fat pad weight expressed as a percentage of the body weight and what you can see here it's a very small increase but in this experiment it was significant you can see a significant increase in abdominal fat pad weight with this necrotic enteritis model so very different from what we've seen with a coccidia model where we're really pushing that energy loss and we actually saw a loss in in fact composition we've actually oh I didn't want to come back to the digestibility and just reiterate this if you look at all the cellular damage that occurs in the intestines and this there's got to be nutrient digestibility losses here but unfortunately this is something that I could not find I think there's been some work on I just couldn't find it in the literature so hopefully we get some reported values on this here soon but again I think this goes back to some of the challenges we have working with this model and we need to continue to work on this model develop this and and generate some of the background data that we have with coxy so that we can understand more what this Clostridium is doing from from a host standpoint so we do have some body composition responses with a Clostridium challenge and as you would expect here so here was a total mass of the birds that we looked at from a DEXA standpoint and then when we look at it from a fat mass standpoint and a lean mass standpoint we can see very very similar reductions so contrary to what we saw before with with coccidiosis it looks like this is a more one-to-one reduction lean mass to fatty mass here and there's not that real challenge to two fat digestion here so just to summarize what we have from a necrotic enteritis standpoint caused by secondary you really need a primary insult bacterial spate species that invades the intestinal tract reduces performance overall if it's a bad outbreak it's certainly going to be accompanied with increased mortality invasion of the intestines causes that immune response has been talked about earlier little information out there on amino acid and energy digestibility but doesn't seem to have as large effect on fat metabolism or it might even increase it a little bit but I think those those those values are still sort of questionable in my mind I think we need a little bit more info from that standpoint but from a body composition standpoint we certainly see that reduced performance and that's partition apart across both that lean and fatty tissue pretty equally and then in conclusion in the absence of antibiotics both coccidiosis and necrotic enteritis will continue to be a major economic drivers and the antibiotics here I'm talking about are mostly the ionophores certainly necrotic counter itis can be controlled with with antibiotics as well but again as that coxy is is really a precursor to necrotic you know the removal of ionophores I think is going to be very difficult to overcome although both can attack poultry in similar areas and they can they can facilitate cross infection the metabolic responses of these birds really do seem to differ and the coxy appears to alter that fat metabolism more and you see increased issues with with fat absorption and even longer term issues with fat metabolism in general and then that the necrotic actually appears to be relatively consistent across both that lean tissue and that fatty tissue so and the body composition data do seem to validate these conclusions so certainly thank you guys for your attention and if I'd be able to answer any questions [Applause] [Music]