good morning good afternoon good evening ladies and gentlemen it's a great pleasure to me to join this webinar and talk today by my favorite subject mycotoxins and oxidative stress i have been working with my antioxidants for the last 40 years and about 20 years were also associated with mycotoxins and today it's a good opportunity to share with you a recent finding so usually i start with this slide just for you to understand that we are living in this dangerous world of free radicals and those radicals are produced in nature and we need some kind of umbrella which we called antioxidant protection we mean sour chicken our pigs our cows everybody need this kind of umbrella when we're talking about uh oxidative stress we need to talk about free radicals what are free radicals they are actuated molecules of oxygens which actually can damage all types of biological molecules and there are internal and external sources of free radicals in biological systems and among internal sources of mitochondria are considered to be major source of free radicals mitochondria which are responsible for energy production but energy production never been hundred percent efficient and up to three percent of oxygen percent of oxygen can escape from electron transport chain and they become free radicals there are also phagocyte cells in the body which are producing free radicals as a weapon to kill pathogens for them free radical production is a way to deal with pathogens there are also some other processes associated with free radical production as well as external sources of free radical production and if you look at this slide what i would like to show you here that we have antioxidant defense system in the body which is dealing with these reactive oxygen species or free radicals and here you can also see microtoxins as a one major source exogenous source of promotional free radical production and what also here important to mention that the physiological level of these free radicals are important for immune defense for cell signaling and many other functions in the body however the overproduction of free radicals it's a damaging effect and at this slide i would like to underline once more that free radicals damage lipids they damage dna they damage proteins in fact free radicals can damage all types of biological molecules and what is important to remember that the oxidative stress as a result of this balance between antioxidant protection and free radical production is a major cause or major molecular mechanism or major stresses in poultry and animal production and if you look at this slide i just summarized main stresses in poultry production including environmental stresses technological stresses nutritional stresses internal stresses and here mycotoxins are considered to be major nutritional stresses which are causing a lot of problems and you already participated in three previous meetings related to mycotoxins where the negative consequences of mycotoxin contamination have been explained so when we're talking about mycotoxins we are talking about silent killers invisible thieves unavoidable contaminants natural toxicons all these names have been given to a group of fungal metabolites called mycotoxins and then negative effects on animal production are incalculable and if we look at this slide this is the most recent one 2020 which summarize the research in the area of mycotoxins you can see here's a 22 percent of research related to aflatoxin 18 to dawn 16 percent zero unknown 15 percent if ocra toxins and combined microtoxins studies are related to 20 of total research and then here you can also see that the research related to mycotoxins in chicken in total comprise about 14 and take 23 percent so almost 40 percent of all research related to mycotoxins is associated with their effect on pigs and chicken and as i mentioned already that mycotoxins have detrimental effect on poultry on pigs on cows on any farm animals and the major mycotoxins of concerns include aflatoxin occur toxin for monasines t2 and dawn and zeralen on and today the major point of the presentation is to consider more in detail molecular mechanisms or microtoxin action with a specific references to oxidative stress and this paper came 2008 so it's already 13 years old at that time we suggested that actually microtoxins are in the world in creations oxidative stress and the inhibition of protein synthesis dna and rna synthesis and damages to dna this related to ochre toxin tetrox toxin aflatoxin and other microtoxins most of microtoxins impose lipid peroxidation and structure membrane damages then we have also induction of the program of cell death which is called apoptosis practically all mycotoxins are involved then we we're talking about transcription factors and gene expression regulation where mycotoxins are involved and finally maintain our disturbances of redox balance in the gut and redux balance in cells are considered as a major consequences of mycotoxin toxicity so as a result we have immunosuppressant hepatotoxicity nephrotoxicity neurotoxicity genotoxicity and uh in this case we are talking about increased mortality poor feed conversion ratio to a gross rate feed refusal decreased fertility and hatch ability more recent paper 2018 confirms the same this is a review paper which again showing the tough latoxin formulas in ochre toxin zeralandon dawn nivalinol t2 toxin they impose oxidative stress they affect mitochondria the major source of free radical production they impose lipid peroxidation they impose apoptosis and they also also affect or stimulate inflammatory response therefore the pro-oxidant properties of mycotoxins are well established and this most recent review which is uh came this year actually uh showing that the major mycotoxins including aflatox and ochratoxin zeralenon fumanizine t2 ht2 and and dawn they are produced uh by uh various phone fungus and we are talking about aspergillus penicillium and fusarium as three major genera in the involved in mycotoxin production and you can see here in various food and feed materials those mycotoxins can be found and more specifically if we look at the molecular mechanism of mycotoxin action which was characterized in this most recent paper you can see that the major research in protein were done with broilers aflatoxin ochre toxin ht2 and t2 toxin were started and here you can see different doses of mycotoxins were used as an oxidative stress in this indications the melon dilde height final product of liquid peroxidation was used as well as a concentration of glutathione major antioxidants and total antioxidant potential and what they showed very clearly that most antioxidant enzyme activity were decreased due to mycotoxin contamination this means that the oxidative stress was imposed if we look at the same paper on the peak side again you can see here dawn zeralenona flatoxin for monizing they also were used in different doses a similar oxidative stress parameters were started including mda glutathione total antioxidant activity and they were compromised and most importantly again antioxidant activity or or an antioxidant enzyme activity we're down like catalase superoxide dismutase glutathione peroxidase these are enzymes which are responsible for antioxidant protection and decreased their activity is related to oxidative stress and here is also interesting that for inflammatory cytokines are increased and what about ruminants yes look here goats sheep lamb cows again the same mycotoxins were tested various levels of mycotoxins and similar to monogastic animals again melon dilde height goes up total antioxidant activity goes down glutathione is a major cellular antioxidant goes down and similar to previous studies their antioxidant enzyme activities goes down so what is the conclusion the conclusion is that for all animal species including poetry pigs and ruminant animals mycotoxins are the problem mycotoxins imposed oxidative stress and this oxidative stress is mainly related to disturbances of antioxidant enzyme activity and this paper which came 2018 is very interesting in a way they look at the effect of mycotoxin sources on the mitochondria as i mentioned that mitochondria are major source of free radical production and if the mycotoxins can actually compromise function of mitochondria more free radicals will be produced and here again you can see aflatoxin dawn fulmonazine ochre toxin t2 toxins zeralanone and they actually were tested in various modal systems and it was very clearly showed mitochondrial dysfunction due to mycotoxin contamination so this is one of the possible mechanisms of toxic effect on mycotoxins affecting mitochondria by affecting mitochondria free radical production is increased antioxidant defenses are compromised and we are talking about oxidative stress and most recent papers actually have more and more attention paid to two transcription factors these are specific factors which activate genes so one transcription factor called nrf2 this transcription factor is responsive for synthesis or activation of synthesis more than 200 of different molecules most of them are antioxidants and aflatoxin dawn or toxins they affect the expression of these transcripts in fact and this effect is those and exposure dependent in the ways that low doses and short exposure usually activate nrf2 which means at the beginning the body can try to protect itself from a toxic effect of mitochondria microtoxins while higher doses and longer exposure inhibited and left to expression and another transcription factor called an af copper b and this factor is very important because it is responsive for synthesis of pro-inflammatory cytokines and pro-inflammatory cytokines impose inflammation in their gut in other tissues and you can see here data on on the effect of dawn and other mycotoxins really known on the expression of this nf kappa b look at this paper in particular it's 2010 paper and what is shown here that an f kappa b expression increased almost six times uh uh so in this case dawn can substantially increase the expression of this transcription factor more preflammatory cytokines are synthesized and this explains why dawn is so damaging on their god now effect of mycotoxins on an f kappa b expression is condition dependent depending on type of microtoxin those duration of exposure they can activate or inhibit an f kappa b and most importantly there is a balance between between these two transcripts and factors and this balance is very much determined redox balance in the cell therefore what are consequences of redox balance disturbances by microtoxins i mentioned already lipid and protein oxidation by the way more and more attention today is paid to protein oxidation because lipid oxidation that something uh described many years ago but uh in recent years more attention is paid to protein oxidation as a major cause of negative effect of mycotoxins apoptosis which is a program cell there's leaky gut and microbiota disturbances immunosuppressions damages to healthy tissues including liver decreased productive and reproductive performance and this few papers just to confirm the negative effect of mycotoxins on apoptosis so mycotoxins in including fumanizines t2 toxin and the other fusarium microtoxins they easily impose a program cell death which is called apoptosis and here one of the paper and you can see here the tito toxin increased the activity of caspase-2 this is a specific enzyme responsible for activation of apoptosis you can see increase in four times eight times 12 times so the activation of apoptosis is very high one more point and again one of the recent paper considering compromise intestinal barrier induced by microtoxins so microtoxins can actually compromise microbiota in the gut compromise chemical by a physical barrier and immunological barrier in the god and that's why mycotoxins very often are associated with various diseases because the protective effect of the god is compromised so when we're talking about healthy gut we are talking about digestion and absorption protective barrier immunity and balance microbiota you can see here normal villis which are responsible for absorption of nutrients and these are damaged relies which could be due to mycotoxin do not expect good absorption and good conversion feed conversion ratio in this case so free radical since they got produced by mitochondria phagocytes and various enzymes and today the actual topic redox balance is a god is a very fashionable one you can see plenty of reviews in a recent couple of years and redox balance and microbiota another topic helping to explain how mycotoxin can affect their health in particular mycotoxin can compromise microbiota and in general terms if we look here the syndrome of malabsorption that's something which is uh considered to be a common for aflatoxic causes t2 toxic causes or toxic causes and this is a syndrome malabsorption is a result of damages to their god so what we have here we have mycotoxins in the feed stimulation of lipid and protein oxidation intestine oxidative stress and damages to parasites nutrient absorption decrease and toxin system is compromised at the same time mycotoxins are absorbed delivered to target tissues and metabolized and oxidative stress is imposed so we have detrimental consequences for health and generally speaking antioxidants actually can help to deal with this problem and they can help with gut protection with liver protection and immunity maintenance therefore the whole idea is that previous first part of my presentation proves the point that microtoxins in the diet can actually impose oxidative stress which means there is a disbalance with a risk between a free radical production and antioxidant differences on one side microtoxins increase free radical production for example by damaging mitochondria on the other side they decrease antioxidant differences due to decreased activity when torques and enzymes and this is related to few transcription factors including nrf2 which is responsive for synthesis more than 200 of different antioxidants and an f kappa b the specific uh transcription factor responsible for inflammation and let's now on the second part of brazilian attention let's look at nutritional antioxidants and protect as protective strategy against microtoxins we need to remember that mycotoxins are very damaging even if you use some mycotoxin binders you are never be able to bind hundred percent of mycotoxins those who which are escaping binding they are coming to the liver to be detoxified and the detoxification in the liver causing oxidative stress and damages to the liver so we need liver protection on the other side dawn is detoxified in the gut and gut microbiota are also can be damaged due to mycotoxin contamination and we need to maintain healthy god so when we're talking about antioxidant protection we need to remember that all antioxidants in the body are working together as a team we call it vitamin e headquarter one two ox and defense carotenoids communicating services of antioxidant defense flavonoids and polyphenolic antioxidant police we are talking about vitamin c and special forces selenium chief executive antioxidant defense and here we are talking about minister of defense which is gene expression and synthesis of various antioxidant enzymes so when we're talking about nutritional modulation of antioxidant system we can consider selenium vitamin d vitamin c polyphenolics and other nutrients which actually can affect antioxidant systems and by improving antioxidant defenses they can help to deal with mycotoxin contamination so i will start from selenium because it's my favorite subject and i have been working with selenium for the last 20 years today we know that selenium is a part of cell of 25 celino proteins those celino proteins are synthesized in the body in response to stress and at this slide you can clearly see that more than half so from those 25 celina proteins are involved in in their antioxidant defense and signaling and now we are talking about dietary selenium which can affect antioxidant defenses it's our review in 2019 and in this case we are talking about breeder egg embryo diocese postnatal cheeks they are protected due to dietary supplement selenium supplementation if we look at the balance between an f kappa b and nrf2 in the gut you can see that mycotoxins pathogens and various stresses can impose oxidative stress and dysbiosis and in this case an f kappa b and nrf2 can be affected in this case you can see the nf kappa b is increased so the synthesis of brain inflammatory cytokinesis increased and nf nrf2 is decreased which means that antioxidant defenses are decreased so if we look further we will be looking at inflammation apoptosis immunosuppression gut structure damages now what happens if we have optimal selenium status so optimal selenium status will help celino protein expression so there will be 25 celino proteins synthesized then we have redox balance regulation uh improvement of antioxidant protection and this antioxidant protection will affect or prevent this oxidative stress imposed by microtoxins then we have effect on prevention of activation nf kappa b which means prevention of inflammation and improvement of antioxidant defenses and we also have maintenance of commensal microbiota as a result we have activation of nrf2 activation of vita genes and decreased activity on f copper b and finally maintaining redox balance gut structure immunocompetence and gut health so here this selenium is taken as a just as an example so in general terms the antioxidants in the gut can help can help to maintain these transcription factors in optimal activity can help to maintain integrity of the god and this will help with first antioxidant defenses and secondly prevent the actually penetration of pathogens through the god and here the review of 2018 and using this review we can show how the system works so we have oxidative stress imposed by mycotoxins the disruption of epithelial barrier is taking place bacteria penetrates this barrier as a result the dendritic cells and microphages will be activated microphages started to produce reactive oxygen oxygen species which mercury radicals and they disturb redox balance as a result we you can see mitochondrial dysfunction which means that free radical production will be increased even more and what we have finally we have leaky gut leaky gut means that the barrier is disrupted and the situation is very difficult and in this case we can talk for example about syndrome or malabsorption so on the right side look what selenium can do selenium can affect the prostaglandin synthesis and activation of various transcription factors which actually prevent activation of this nf capability which is responsive for inflammation so the pro pro-inflammatory cytokines goes down anti-inflammatory goes up we are talking about additional synthesis of selena proteins we are talking about additional synthesis of antioxidant enzymes like superoxide dismutase and catalase and what is happening having here we are talking about redox balance reestablishment and inflammation resolution so this means that inflammation is taken under the control and in this case we are talking about maintenance of the gut structure and antioxidant defenses in the gut uh will keep healthy god and prevent those damages uh caused by mycotoxins so now when we're talking about selenium we need to remember that there are different sources of selenium and there is an inorganic selenium which is its first generation of supplements which came in many years ago and in this case the this sodium selenite is used in many commercial premises but the efficiency is low uh second generation of supplement include serena yeast and pure selena methionine and they have some problems like selena methionine consent proportion in selena yeast only 60 70 percent and we have also some problem with stability issues pure selenium methane so finally the such generation of supplements came on their market which is hydroxyl selenium methionine helping solving selenium problems and helping to maintain optimal optimal antioxidant defenses and this can help with mycotoxins okay that was a selenium however the most important antioxidant and most study antioxidant in biological systems is vitamin e so vitamin e is antioxidant which is located in cellular membranes and vitamin e is preventing damages to those membranes and if you look at the our review 2019 you can see that vitamin e has protective effect for breeders and cockrells for semen for egg yolk for embryo and for newly hatched chick and you can see here vitamin e increase antioxidant defenses not only by increasing vitamin e itself but also by activating superoxidase materials glutathione peroxidase and catalase you remember from my previous slide the disease enzyme activity was actually decreased due to mycotoxin contamination so in this case by using vitamin e we can re-establish defenses in stress conditions do we have any evidence related to microtoxins yes if you look at this publication in 2008 it's a effect of t2 toxin and dawn on dna damages so you can see here dawn and t2 toxin impose dna damages and by adding vitamin e you can decrease those damages and if we look more specifically in the review paper 2018 which actually look at the protective effect of vitamin e again is mycotoxins we are talking about alpha toxin dawn for magazine orchard talks and t two talks and zeralinone in all those questions cases of vitamin a has a protective effect again protecting against oxidative stress protective against uh membrane damages against uh are actually this activation of antioxidant enzymes what about vitamin c another antioxidant again you can see in the same reviews they showed that aflatoxin dawn ochre toxin keto toxin and zeralinone they also they are negative consequences also can be mitigated by using a vitamin c and again you can see increase in glutathione concentration glutathione peroxidase and superoxide dismutase activity those enzymes which are inactivated by microtoxins decreased mda level which means decreased lipid peroxidation and again in previous my slides you saw that mycotoxins actually negatively affect uh lipid peroxidation increase lipid peroxidation the vitamin c can help you and finally the most exciting part it's a polyphenolics the polyphenolics there are more than eight thousand compounds in nature uh you can see there are flower flavonoids flavanols flavanones proventations and other group of these compounds and in general terms polyphenolics received a lot of attention in the last 20 years and they were considered as important antioxidants and more and more data accumulated that the action of these polyphenolics is related to those transcription factors i mentioned already so do we have any evidence that that they have protective effect in microtoxic causes so if you look at this big study which is 2019 and you can see here that the activity of catalase glutathione terexitase and superoxide dismutase in duodenum in the gut was compromised you can see a significant decrease and by adding grapeseed extract which is a source of polyphenolics this was re-established which means that by using polyphenolics we can maintain antioxidant defenses at the ordinance the same is true for the liver you can see here again in the liver the protective effect of polyphenolics is obvious another paper 2019 the effect of polyphenols and vitamin e on antioxidant status and meat quality of broiled chickens fed naturally contaminated with ochre toxin a again you can see here the same parameters antioxidant enzymes total antioxidant potential in the blood in the liver in breast muscles and again occur toxin compromise those parameters and polyphenolics and vitamin e actually having protective effect and in some cases the protective effect can be in combination in higher than in individual antioxidants this means that by using these polyphenolics in the diet of chickens it's possible to maintain antioxidant defenses in the gut that's the place where actually damage is most important as i showed you previous slide for example leaky god at the same time uh is a god is responsible for drunken uh detoxification and again to maintain gut integrity to maintain the microbiota it's a very important task and this kind of compounds dietary compounds can help and finally one more compound called silimarine which is a plant extract which is used more than two thousand years in human medicine and this is our big review 2015 which actually described antioxidant properties of silimarine and silimarine is very popular in a way showing antioxidant protection and the in the same at the same time uh you you can you can see here that uh silimarine actually affect the toxicity of aflatoxin so you can see here uh the aflatoxin they were used in chickens and you can see body weight 20 day 28 and day 35 you can see significant decrease and by using silimarin it was recovered the same is true for feed conversion ratio again you can see control group aflatoxin be a contaminated group and protective effect of silimarine and again silimarine is the most effective compound to maintain antioxidant protection in the gut to activate a transcription factor nf2 which is responsible for synthesis of antioxidants enzyme and down regulating nrf nf kappa b transcription factor responsible for the uh inflammation uh one more paper again quite quite recent one uh aflatoxin and taking ducklings effect treatments with lycopene and silimarine you can see the protective effect and finally whatever what about other botanicals i came to the end yes the other botanicals can also help to maintain antioxidants defenses including rosemary baldo and others and one more point probably the last one bht bht is commonly used as a feed additive to deal with a lipid peroxidation and in this case the bht can also help to deal with mycotoxins so conclusions mycotoxins are major nutritional stress factors in protein production ketotoxin don xeralinone ochre toxin aflatoxin and phenomena can impose oxidative stress oxidative stress is major mechanism mycotoxin action and as a result of oxidative stress we have got integrity damages got microbiota damages immune system damages and various organs can be damaged as well this could lead to decreased production and reproductive performance and what antioxidants can do until dietary antioxidants can help to maintain antioxidant protection in the gut antioxidant protection in the tissues and the dietary supplementation of dietary antioxidant like selenium uh some photogenics or botanicals can help to maintain antioxidant protection and can help to deal with mycotoxin toxicity and at my last slide and i thank you very much for your attention okay thank you um we can now get started to the question and answer session we have about 15 minutes for that um please type your questions uh on your chat box there it can be in portuguese spanish english we can definitely translate to the moderators they can uh they can answer questions for us um let me start here with one from ecuador the use of polyphenols can reduce the partici participation of vitamin e on the diet what would be the recommendation i'll send that to dr julia dvorska okay uh i think that um well antioxidants they are as as peter showed in his presentation they're like group or team of antioxidant different antioxidants and they all work in the different parts of the cells for example vitamin e um it works in the cell's membrane to uh to kill or to deactivate the free radicals and but for example after uh the activation of free radicals there is a hydroperoxide which is also need to be deactivated in this case we need to proximity so we need selenium for that and polyphenolic they are acting uh in protection of um mitochondria mitochondria uh protecting from um like keeping the integrity not to produce uh additional free radicals due to mycotoxin mycotoxin action so they are protecting my mitochondria and decreasing the also um production of free radicals so we need already three of them so i want to say that i just wanted to say that antioxidants they are working with tea so we need all of them it's not enough only to use vitamin e and to support to maintain physiological needs of of the body and at the same time to deal with the stress stress condition on oxidative stress due to mycotoxic causes for example so in this case we need all of them for different purposes so okay thank you i will send one to dr olga brakiva now what's an acceptable level of crno in raw materials like corn when a raw material for high level my toxin is used to mix feed to these mac toxins get diluted when a small amount is used in mix and feed i get it um it depends of course on the country because there are different acceptable levels uh in different countries and are we talking about physiologically acceptable levels or governmentally acceptable levels normally government already issues some guidelines for uh main raw materials especially for the area known that will be wheat maize soya and they could be quite high especially if this grain is intended for the animal use but if we really see the levels that are allowed or guided for the feed they are not really correlating with the inclusion rate of this grain again if i go further to physiologically safe levels they are much lower than normally accepted by the government by fda or european community um and of course depend on the species so uh in case of sierra the most sensitive species are mammals so ruminants and swine and less sensitive species are poultry so i can only refer to some european levels by hard but still the physiologically safe levels are much lower and another thing that if we relate your question to the topic of presentation today oxidative stress uh the good news is that sierra zeraliano is not really a very big problem if we talk about the triggering oxidative stress so oxidative stress is mainly caused by trichotosthenes aflatoxins or her toxins so if i would pick up just a species that could be a good acceptable levels for swine i would say if you have 50 microgram per kilogram in the total feed for swine and for aluminums that is really a safe level if you go above 100 microgram per kilogram for the total ratio that could already pose issues in reproductive animals in poultry we can go much higher because sierra leone is converted to less estrogenic metabolite thank you um next one for dr dvorska what is the best strategy of binders that address my toxins directly and additives that support and protect natural antioxidant systems um well when we um talk about mycotoxins first of all we need to try to eliminate them from the body as much as possible like we are trying to bind the mycotoxins which are able to be bound like aflatoxin polar for example or ergot alkaloids and then um it's it's good to support the organs like uh like peter already mentioned that mycotoxins goes to the liver for example to the to be deactivated and induce the oxidative stress so if we can protect in this case liver or gut or all the um and also to help antioxidant system to to combat this oxidative stress that will be beneficial but first of all we need to try to eliminate mycotoxins and then to support all the organs including um including antioxidants since they are like their levels are decreasing due to mycotoxic causes and it's not enough to to deal with mycotoxins okay thank you i will uh just uh get another question on this same topic or translating here in spanish what's the best combination or the correct combination of all these different antioxidant products to help with this uh issue combination of antioxidants right yeah these they just had uh correct of all these different products or yeah antioxidants or different ingredients that you can use as already mentioned by peter uh selenium can be uh very good um additional solutions when we have an anti-mycotoxin anti-mycotoxin products and then to help uh antioxidant system to to deal with microtoxins we can use selenium as a my main part of my of antioxidant defense but then also as he mentioned very popular now it's polyphenolic compounds from different botanicals like silimarine or other compounds they also they can help to keep integrity of mitochondria and decrease decrease their free radical production by by mitochondria due to mega taxi causes so again we can use different combination of uh botanicals to to get polyphenolics and uh yes vitamin e we have already in premixes so it's no need to increase level of vitamin e since it's all we have already high levels and could be vitamin c could be added to the diet especially during the heat stress but only in this case not not not just for mica toxic causes for example yeah guillermo is frozen i believe yeah okay so mateos you can read the questions then yes let's see other questions and is the oxidative stress the the mycotoxinal result of right production of free hydrical production or a decrease of endogenous antioxidant defense it's probably like i i can address also this question from from the previous one so when micro toxic when mycotoxin induced oxidative stress first of all they are increasing production of free radicals by mitochondria so and then at the same time they are decreasing the antioxidant difference so both of them and both of them uh it's the mechanism of mycotoxicos of pro-oxidant action of mycotoxins in the cells so the answer is two of them both good and we have other and i think it's a good question to oga would the standard oxidant levels in feed enough to protect the animals against the ox the team stress if is already using bha bht or other antioxidants a good and very relevant question i hear it all the time from the customers and indeed that some synthetic antioxidants like bha bht could be uh taken in vivo could have a potential benefit against oxidative stress and vivo but this is not their uh purpose of the use and normally uh the level of antioxidants like bha bht i will call them feed antioxidants is [Music] dosed or is chosen according to the oxidative status of the feed so that means that feed oxidative status will uh impact on availability of those antioxidants so they will be consumed by free radicals which are present in feed so of course if you have excess of bha and bht in the feed it's possible that the the body systems oxidative systems will take it for themselves for any in vivo oxidative stress but in most cases uh the level is not enough it's consumed by feed free radicals already in the feed okay and we have other interesting question what type of mycotoxin binders binders is recommended to minimize mycotoxin exposure and its effects on oxidative stress what type [Laughter] yes julia is our expert in oxidative stress she has already several publications and sometimes i would need her help in answering questions so the complex mycotoxin binders or i would say deactivators would be the best solution because oxidative stress is a kind of the consequence of mycotoxins and even if you use a good binder which is claiming that they bind high levels of mycotoxins still in the nvivo systems it's not possible to bind hundred percent of any mycotoxin even aflatoxins which is very easy to bind so always that will be a limitation some mycotoxins will escape from binding and will affect the immune system and oxidative systems and i would strongly recommend not to count only on binders if you would like also to manage mycotoxin issue including oxidative stress caused by mycotoxins that the best to use some compounds like peter mentioned in his presentation some nutritional strategies to repair and to uh counteract with negative consequences of those mycotoxins which cannot be bound so this is a combination of selenium of vitamin e vitamin c sometimes even vitamin e can be involved and of course the use of some botanical compounds and curbs with proven efficacy to maintain um oxidative stress is also quite beneficial and this is one of the let's say cost-effective solutions agree with organ especially especially now when vitamin prices could go high and sometimes really going very high that could be effective maybe to adjust a little bit nutritional strategies according to the profitability results yes and during the stresses the requirement in antioxidant is increasing and micro mica toxicosis is also oxidative stress and like uh requirements are higher so the inactivate of the activator of mycotoxins this additional antioxidant would be beneficial in this case like to increase these requirements or requirements during the stresses okay yeah sorry about that i thought my internet dropped here for a few minutes thanks mattels for taking the lead on uh for a few minutes and i will follow up with some questions here um can animals be experiencing oxidative stress caused by my toxins even when they have no clinical signs of my toxicosis um one of one of you two ladies if you want to answer we can we can answer together i can start um yes animals can be uh already under the stress even if we don't see even any clinical science because the stress conditions that are happening on the cells levels like people already showed us uh like damaging of membranes of cells and in especially in attached organs for microtoxic mycotoxins for example in the liver damage of cells in the liver or in the guts and also it can lead to apoptosis program cell death is also one of the mechanism of peroxidant action of mica toxic mycotoxins and also in in the gut we can see all the on the damages from don for example on the cells level and then we can see the leaky gut or malabsorption syndrome or anything so even with if without clinical signs stress oxidative stress can be already happening on the cells when we have mycotoxins in the feet and i would add from my side that oxidative stress is a beginning of the problem so when you don't see it yet doesn't mean that it will not happen so in fact when you measure oxidative stress or you are able to control it or see it at the early stages you can prevent a lot of diseases which will be a consequence of oxidative stress so yes oxidative stress begins first but uh often i think in all cases it's not really seen by veterinarians or by um people who take care of animals okay thank you i think uh we are now on top of the hour it's been an hour that we started uh i appreciate the moderators for the question and answers and i appreciate all the our partners in the in the industry to join us to join us today for this webinar and i hope uh we can see all you again on may 25th for the for the final episode on the michael info series so thank you very much and please uh reach out if you yeah if you have any further questions we would be glad to answer them thank you bye-bye thank you thank you bye-bye