okay thank you very much to the organising committee here for inviting me to give this presentation so my name is Charlotte Louis and I'm from ojos University in Denmark and my presentation is entitled from oxidative stress inflammation redox balance and immune system so most of my research has been really focusing on pigs but since its poultry conference I'll try to draw in poultry as well so the concept of my lecture today is as a starting point to show for you that we have some major challenges in pic and poultry production then I'll continue to try to emphasize when and why livestock may suffer from oxidative stress and inflammatory reactions and then I'll continue on the dietary inference and redox stasis of the animal I will focus on fatty acids and immunity and then anti oxidant deficiency and immunity and supplementation of antioxidants and immunity and finally some conclusion and perspectives so the challenges in our pig production you may know that Denmark is really producing a lot of pigs many more pics and people in Denmark and we have kind of spread out our genes towards the world so we are having these hyper prolific sounds and they are giving around the twenty piglets per liter and but these leaders they are very heterogeneous and we are facing a high piglet mortality so this is a critical phase of life but also weaning from the South is a stressful period for the piglets it has to adapt to new feet and environments it is getting separated from the dam which is also a psychological stressing factor and during this period the post weaning period that is really an increased risk of enteric diseases which means that we have a high consumption of antimicrobials during this part of a piglet life and in Denmark we are currently using instead of antimicrobials the super nutritional levels of sync but from an environmental perspective this is not a good thing to do so today we are really looking for alternatives to antibiotics and sync so this will come fee a starting point of our research today is really how can we improve resilience and robustness of the piglet just to demonstrate what I have said so far in some numbers so we have this survey 10 map you can look it up this is a yearly report and the use of antimicrobial agents and occurrence of antimicrobial resistance in bacteria from food animals food and humans in Finnmark so already in the 90s we gave up using the the antibiotic growth promoters and since that time this was actually before the ban in in the European Union so and we have reduced this but also you can see that there has been a steady increase in the use of prescribed veterinary antimicrobials still we are producing more and more pigs in Denmark so today we are really saying what can we do to reduce this antibiotic usage and what we have today is this yellow card initiative given to the farmers if they are using too much the thing is now that the majority of the answer microbials in the pig production that really goes to during the post weaning period around 85% of and antimicrobials is really being used for treating and for enteric diseases in the post winners recently we have had this debate on the increase in veterinary microbials because of the matzoh resistance bacteria and now we also put in focus and medical sync so this is just to show for you that the diarrhea that's the frequent observed a gastrointestinal illness sin takes it the most out perks occurs during the second week post weaning the post weaning diarrhea what BTC can be really high up to 50% and mostly the the cause is inserted so genic ecoli stomach ulcers is also an important inflammatory reaction but so far we are not really sure about the impact of bacteria here maybe it's not the cause but a contributing factor what you should see from the pictures here really at clear clinical signs of inflammatory reactions during these diseases also just to show for you the problem with the sink the consumption of zinc oxide the Supra nutritionally also sings has increased and this is really because we couldn't use antimicrobials so far anymore this is critical and we have to do something we also see the challenges in the poultry production I should say now that Denmark is not really main producer of poultry so this illustrates also here that this segment of the livestock production is not the major contributor to the use of antimicrobials what we use of the ants microbials for in the poultry production is the treatment of coccidiosis this disease is characterized by oxidative stress inflammation diarrhea nutrient malabsorption fluid loss and dehydration so there are other inflammatory reactions and frequently observed diseases umbrellas the dis bacterial lysis it's not a well-defined inflammation of the small intestine but it occurs in the period 21 to 28 days and we have the new curtain rice rin sorry necrotic enter Isis which is a severe intestinal inflammation occurring almost the same time and here we have the major a contribution by the toxin producing frustrated by thinking's here now these infections are and we have in my research group and major focus on gut health and disease prevention now these infections trickles and inflammatory response it's basic knowledge here that the bacteria trigger macrophages to release cytokines and chemokines we have during this process of the release of inflammatory mediators later and you see the vasodilation and vascular permeability and then the influx of phagocytes fortunately this is a reaction going on it's a natural host mechanism to combat will invading pathogens one is now the length of infectious disease to oxidative stress well there was a question here about what are the sense accidents just we've seen so much today by the former speakers about oxidative stress my is the imbalance between the production of the racks of parasites and Jhin radicals and then the scavenging ability of the hooves to destroy them so the question is now when do we or now the animals produce reactive oxygen species it's not only during the elimination of foreign material by immune cells it's actually a part of normal metabolism as also demonstrated by walls of this during the first presentation but the high genetic potential under intensive farming conditions may actually elevate the production of these rates of oxygen species here you can see again how the bacteria being engulfed by the facts I ptosis which is now really and how the the bacteria is going to to cope with the invading bacteria but the very important mechanisms by which the macrophages and other immune cells are going to kill the bacteria it's just illustrate here that these oxidative rates of oxidant species they are so important to use for and immune cells to kill the bacteria these are the weapons and as we have clearly seen that the most the highest production of reactive oxygen species are going on in mitochondria and the thing is if we can't control this it may lead to mitochondrial dysfunction and this has actually been proven especially in human diseases to be linked to watts of disease progression I think it's very interesting with some recent research carried out that also the genetic factors such as intensive genetic enhancement seen as the production cycle for broilers has been shortened to 35 days and this can actually also be related to an increased production of reactive oxygen species and also the improvements or you can talk about improvement but at least genetics two or more guinea pigs but these pigs they have abnormally high serum concentrations of four active oxygen species as opposed to rules why when it means actually that you have oxidative stress conditions during resting similar to that of human during intense physical training so it means we should really keep an eye that this genetic progression may lead to a higher level in general of oxidative stress now going further the redox balance and immunity it is possible to actually buy nutrients neutral neutral modulation and to enhance the resistance to infectious diseases and this is especially placing who has been here maybe still is around has demonstrated that deficiency of fatty acids vitamins and selenium may actually lead to damage to the development of immunity and the direct regulatory actions of these nutrients so I have during my research asked myself how can you actually buy that sort of manipulation with fatty acids vitamin E is what kind of mechanisms and effects would it have an immunity in the intakes so what you can see from this figure made by Professor glazing is that you want to be here in the I don't know if you can see it yes you can see the red spot here that and this is really the region you would like to be in you don't want to be doing a deficiency system and situation or toxicity both would actually influence and the immunity of the animal towards a critical situation we have not discussed that much today about fatty acids but the singers that these inflammatory reactions they are really two really related to the level of arachidonic in some membranes a phospholipid region so once of arachidonic acid is getting liberated by phospholipase a2 it produces press Atlanta in into and this is performed by the sacra oxygen enzymes and these this process is important not only in inflammatory reactions but also for wound healing but here you can see that in parallel you can also have the production of reactive oxygen species everything here is a natural and important reaction but if it gets out of control and it is really enhancing the disease progression and this is why it is so important to try to control it all just in a few moment demonstrate how this is possible by a fatty acid manipulation and then here how you can also use antioxidant therapy to inhibit these rats of oxidant species a production it's really interesting will the research performed where you have combined n minus 6 and n minus 3 fatty acids it is very easy by dietary means to manipulate the cellular composition of fatty acids and although if you see here that from the North acid you get a production of arachidonic acid and this as I just show for you will actually produce eicosanoids which are heavy induces affirmation of inflammatory reactions but here if you now take the alpha linolenic acid that you can by chain elongation and this saturation also have a production eicosanoids and although they're very similar in molecular structure they are less important initiators and of implement inflammation so here is to show for you that we can actually play around with the ratio between and c18 3 and c18 2 in the diets and often we would our we've done a lot of research to see if we can actually influence the production of the longer chain polyunsaturated fatty acids by inclusion of more c18 free but it is not that easy just recently we performed a study with the hemp eye versus soybean oil unfortunately was not the funny hemp here it was industrialized him and we produced oil out of this and although when fat you know him for soybean oil you see here that you change the ratio between in six and then free and higher in soybean oil though for him by but although these cells which we sample blood we sample blood and milk and they were fed him soil and oil without any of the longer change of heart and seven feet fatty acids EPA DHA and so we were not able to detect anything in the also the feet but these fatty acids were presents in colostrum and milk and Jasmine of South and in the piglets plasma so as also demonstrated in other research studies these fatty acids are important for the piglets for instance in order to fall for the mucosal immune responses to appear and I think that maybe we should keep in mind that the fatty acids are probably those which are the biological most important signals for the immunity the answer oxidants are not behaving in the same way but they should again they're there to control the oxidative stress reactions but the fatty acids are those who actually influencing cytokine production and all the other signal molecules important for the immunity there are many ways by which nutrition diets can modulate incisional ecosystem this fatty acids of course are only part of it but I think one of the major important things are that you can directly influence the composition of the immune cells in the intestine some of you know the nutritional products prebiotics probiotics may be they they changed more than microvolts and I know there are a lot of studies now addressing probiotics but again interestingly the fatty acids are those which have a direct influence on the immune cells here so we have since we know that it's not possible to get the epa DHA and produced by inclusion of vegetable oils rich in c HC nancy free we having occluded fish oil for in piglets studies and that you can see that if we now study the fatty acid concentration of the god my my my closer in 56 days old pigs that's four weeks after weaning we really see a changed fatty acid profile of the god Lucas are here and when we now start to put a challenge on the immune cells here its macrophages harvested from the lungs this is more eases and harvesting macrophages from the guts and you can see that you can actually influence and the production of eicosanoids here at the PG 2 and actually the fish oil to the wieners inhibited the production of PG 2 in comparison to the sunflower and also animal says but of course you should keep in mind that once you start to play around with the fatty acids in the diets for the animals you can really influence also the susceptibility to oxidation because oxidation rate depends on the number of double bonds and you can see you have affected from one to 2004 going from C 18 up to C 18 free so this is why we need to control this reaction now when we start soon put a challenge on the animal we actually the immune cells and once they are going to be called for they change in order to increase their antimicrobial efficiency and development of immune response this I demonstrated later earlier on you see this increased production of oxygen derived products so the dietary antioxidants they can actually help to limit collateral damage by the free radicals to the host tissue now you have already heard a lot about I'm happy about that's because vitamin E is still my eyes most important biological and oxidant it is sitting there in between the fatty acyl and fatty acyl in the cell membranes and it prevents the oxidation of the fatty acids in the cellular membranes and the way it does that is to donate hydrogen and to the perks of radical in that way you terminate their reaction now we have at our Institute several animal models one of them is now ecoli inoculation two picks and interestingly enough you can see here that irrespective of the vitamin E level in the feet is was either given a control level or a supplement level you can see that once you start to challenge pigs will ecoli you actually reduce the vitamin E concentration in the liver of pigs by approximately 30% recently we in this hemp study we have confirmed these results and we actually observed a reduced plasma vitamin E concentration in pigs after an e.coli inoculation so what is going on again I think that once all these processes I said to you that once the invading bacteria come into the intestine there is this reaction of reactive oxygen species produced and then there is a need for more and succeeding protection otherwise the God's issue would be destroyed fortunately there are not only one antioxidant vitamin E but are so of them already we have heard about the closest iron peroxidases is to worksite disputatious vitamin C and so forth and this interaction you also heard about by vitamin E and glutathione peroxidase this is a bit simplistic because glutathione peroxidase we now know it's not the only one we have 25 Salina proteins and again there is a nice collaboration between vitamin E and then the more for water sobel the in somatic antioxidants sitting there in the water phase of the membrane and in that way they can scavenge and terminate the out oxidation processes now when is it actually a critical situation when do we have vitamin E and selenium deficiency and what is now the link to immunity it has been observed in from long time ago that when Atty was depleted in vitamin E and selenium you sore suppressed lymphocyte response nitrogens compared with vitamin E and selenium depletion alone again it was set by this ad here that the key may be the fatty acids suppression of the immune system might be attributable to oxidative products from fatty acids antioxidants may prevent inhibit proliferation of lymphocytes and alterations in the moon cells the critical face as I said in the beginning of my presentation is really during the post weaning period of a piglets life because while it's suckling the sow it is getting a lot of passive left Rajini immunity factors once we win the piglet there is a steady decrease and since the piglet is weaned may be too immature it is still not having an active immunity we have seen this in one of our studies that actually already at they fall after birth I mean the the piglet the pig is spawned vitamin E in a deficient but four days after and the sow has transferred so much vitamin E through the colostrum that the vitamin E status is built up then we win the piglet and you can see that vitamin E is really decreasing here this has to do with the limited capability of the piglet to absorb fat soluble nutrients and also vitamin E and I was often a limited endogenous synthesis of vitamin C during this period of life so in my eyes and we really we really have to address this period of a piglet life and because this is where you see severe accidents stress condition if you don't have enough antioxidant protection now it is possible to treat the piglets will vitamin E and in that way build up the vitamin E status you can see here we had increasing dietary levels of vitamin E and we follow the pics for weeks after weaning it's defined in the literature that when you are below point six milligrams per liter plasma is a so-called deficiency situation here you have a critical condition and some authors are talking about that in order to have the best immune status of a piglets you need to have at least three milligram vitamin E per litre Chasma however I think we need more results to demonstrate that but based on my own research I would say you can say that an e coli infection really costs in antioxidant concentration measured as vitamin E here so it means that you can easily suddenly be in a critical situation that studies around where but unfortunately you say that they are rather old now and they are not really and on performed on the same production conditions as we have today again the genetic material has changed and also the production systems so although there are some verses showing that increasing levels of vitamin E and would actually enhance immunity of pigs I think we need to again put more focus on this research we have seen that if we supplement south with increasing levels of vitamin E we actually saw an effect on the ants body responds to ecoli of the piglets and also we have seen will pick less wind from South and fed different sole sources of vitamin E but also different levels that vitamin E had no impacts on the cellular mediated immune responses but in that experiment we also had different fatty acid compositions and those were actually the players those were making the differences in immunity so am professor Mahan has contributed a lot to the research in pigs concerning organic and inorganic selenium for south and as already said that the organic selenium forms are making or giving a higher selenium status than the inorganic forms but well the reason why we have focused so much and this from a nutritional point of view is really that in order to have a piglet post weaning will a good selenium status you need to influence the status via the sow feet because hex of pure selenium status encountered the selenium deficiency sooner after weaning as demonstrated by a Meehan and the depletion of selenium reserves will advance in Paris is also something which has been dick emphasized by mayhem simply meaning that increasing parity of the South's would actually put a pressure on the selenium as well so what about supplementation of vitamin E and selenium and immunity it is well documented that alpha tocopheryl stasis can be increased by increasing vitamin E and we have also focused a lot on the differences between natural and synthetic forms of vitamin E and also that the selenium status can be increased by using especially organic selenium rather than in organic but so far very little is known regarding the influence of enhance and subsidence status on immune responses of fixed post weaning when the infectious risk is high so the role of mechanisms of biological important phone Salina proteins to cover all derivatives I think that is important so coming to the conclusion and perspectives what I started out with is that we think we should keep in mind that I've really challenges in the pig and poultry production to use expensive ionic stores infectious diseases that oxidative stress and inflammatory actions are part of normal defense mechanisms against pathogens but also that oxidative stress may be enhanced during intensive production we need to control these productions and the oxidative stress production and also inflammatory reactions to prevent tissue damage during fictions there are several mechanisms which by which we can actually control it and I think overall if we focus more in this optimal and suck Siddhant nutrition may reduce the need for antibiotics by increasing the robustness of the animal towards infection but we need certainly more research on that so finally I'll just like to show for you a picture of our Institute and of Denmark and my my group and if you don't know where Denmark is I probably think you know but you will soon find us in the world map because just close to our Institute they are building up a new Apple data center in the middle of nowhere but that's also where we are situated so with that I like to thank you for listening [Music]