good morning afternoon or evening everyone so as it has been said i'm going to talk about the impact of mycotoxin on immune system so i'm not going to introduce what mycotoxin are because it's not the first seminar but i'm just going to give you an example of the multiple effect that mycotoxin can have and i will take the example in pigs so we know that there is there are different mycotoxin and we know for example that aflatoxin b1 has its main effect on liver it induces liver toxicity but has also an impact on growth as far as oprah toxin is concerned it more targets the kidney dioxin ivalenol and t2 toxin are true trikutican and trichotic and are known to affect growth and feed intake inducing anorexia femonism b1 is known especially in pig to target the lung and the allen on is known to target the reproductive tract but as you see all this mycotoxin have an effect on the immune system especially aflatoxin femonism and trichoticking in fact when talking about the immune system it can be divided into two kind of response there is first innate immune response this respond is rapid but not specific and this innate immune response involves immune and non-immune cell such as epithelial cell endothelial cell and immune cells such as macrophage neutrophils or dendritic cell conversely the acquired immune response is very specific and it is the type of immune response that you have during vaccination but it's slow and the aqua immune response only involves immune cell especially lymphocyte so it's not possible during 40 minutes to give you an overview of the effect of all the mycotoxin on all the type of immune response so i'm just going to give you some example here is the first example that the mycotoxin aflatoxin b1 has an effect on macrophages and as you see on this slide on this part of the slide you see that it reduces the effect on production of nitrogen oxide and about oxygen reactive species in a dose-dependent manner and this lead to an increased cell death of macrophage in the type of apoptosis so we have an effect here showing that aflatoxin b1 decrease the functionality and increase the mortality within some immune cell in our lab we did quite a lot of experiment trying to investigate the effect of mycotoxin on the immune response in peace the experimental protocol that we use was most of the time uh the same the animal were exposed to mycotoxin for four weeks and during these four weeks they also were vaccinated with a model vaccine which is of albumin and during the this four week we look at the immune response both the immoral immune response which is mainly antibody and the cell mediated immune response which is mainly lymphocyte proliferation and we look at this immune response both with for towards the antigen we use the vaccine that we're using but also the total immune response so total antibody or total lymphocyte proliferation and here are some results that we get here is an expert is this experiment where we use t2 toxin and the animal receive either control food or food contaminated with 0.5 1.3 or 2.1 microgram per kilogram t2 toxin during the four weeks and they were vaccinated with this model vaccine of albumin and we look at the antibody response when we look at the total antibody response you clearly see that there were absolutely no effect but when we look at the specific antibody response meaning the antibody response towards the vaccine you clearly see that the group of animals receiving the highest level of mycotoxin had reduced antibody production compared to the control animal or the animal receiving the lowest dose of mycotoxin we did exactly the same response with the same experiment with another mycotoxin here it's athletics in b1 and the animal receive either control feed or feed contaminated with 0.4.9 or 1.8 microgram per kilogram aflatoxin per kilogram feed and we look at lymphocyte proliferation and when we stimulate all the lymphocytes we didn't have any effect but when we look when we stimulate the lymphocyte with the antigen that we use in vaccine you clearly see especially after two weeks that the control animal the animal receiving the control feed responds really well by contrast the animal receiving the feed contaminated didn't respond that way so it indicates that low dose of mycotoxin decrease the antigen specific immune response without altering the global immune response so really to see an effect of mycotoxin on the immune system we need to stimulate the immune system so um as i show you here we look at the immoral and cellular immune response for two mycotoxin t2 toxin or aflatoxin b1 and i show you that for t2 toxin we have to decrease immune where the decrease in immune response especially in terms of antibody production and with aflatoxin we have to decrease lymphocyte proliferation but when we look at the antibody production for aflatoxin b1 there was even the specific antibody production there was absolutely no effect and when we look at the specific cellular immune response for t2 toxin we didn't have any effect so it really indicates that each mycotoxin act differently on the immune response t2 toxin affect the immoral immune response where whereas aflatoxin affects the cellular immune response and so if we need to characterize the immune effect of mycotoxin we need to investigate different parameters of the immune response so to conclude on this first part we've shown that each mycotoxin act differently on the immune response i show you that t2 toxin alter the antibody specific production but doesn't alter the cellular immune response lymphocyte proliferation and this is the opposite effect for aflatoxin b1 so we really need to investigate the effect of mycotoxin on several parameters to be able to see what are the effects of mycotoxin on the immune response another conclusion of this experiment it shows that loads of mycotoxin decrease the specific immune response without altering the global immune response so we really need to stimulate the immune response to investigate the immunosuppressive effect of mycotoxin what is even more important than knowing exactly what mycotoxin are doing on the immune response is to is to study the consequence of the modification of the immunity on pig heads and basically the fact that mycotoxin alters the immune response has two consequences the first is an increased susceptibility to infection and the second is a decreased vaccine efficacy and i'm going to give you some example on these two parameters so here is an experiment that we did also in our lab with piglet that were exposed to feminism b1 and they were after that infected with an oral infection with a pathogenic e coli and what you can see that at the end of the experiment when we look at the number of bacteria in the intestine there was much more bacteria in animal that were exposed to the mycotoxin this is a log scale so the increase was at least tenfold to 100 for more bacteria in the intestine of the animal that get the mycotoxin and here in these photos you can see here the small dots are the bacteria that were stained and you can clearly see the bacteria that are attached to the intestine so this really shows you that the fact that mycotoxin can increase susceptibility to inf to infection this is another example still with feminism v1 but here is in chicken and still e coli infection hits um 21 day old chicks that were injected with e coli and after the bacterial injection they were killed and the number of bacteria were determined in different organ spleen liver or lung and you clearly see that there was um 4.9 10 um in log 10 there was 4.9 bacteria in the control uh animal receiving no uh feminism but uh when they receive feminism it was more than one log one log higher meaning that it was more than 10 fold higher and we see that in the spleen there was also an increase but it was smaller in the liver and there was no effect in the lung this is just for example but if we go through literature there's quite a lot of example that has been [Music] observed and here is just a survey of a literature survey of experiments that have been done in chicken so with done an acetyl form of dawn they are lenient and humanism and with emmeria maxima there was a decrease of cd4 and cda t cell in the mucosal in the mucosa with uh also done acetylated form and then there was um an effect after also infection with emelia there was uh in effect with e coli salmonella clostridia salmonella galinarum or also e coli so there is a lot of evidence that really the fact that mycotoxin altered the immune system will increase susceptibility to infection here's a review on poultry but i can have the same slide on pigs for example this is an experiment that i like to show because um here it's not an experimental infection in this experiment what happened is that it was in a in a form the they were given awkward toxin in the field at one ppm or three ppm and basically what happened is in this form in animal that received the loaders of okra toxin two out of the six animal develop salmonella infection and in the in the group of animals that receive the high dose of mycotoxin they all develop a salmonella infection so he had really indicated that the sanitary status of the farm was not very good because there was sand manila around but basically the animal receiving the normal the control feed didn't develop an infection their immune system was strong enough that they can fight again the infection but when they receive a mycotoxin they could not fight and then they develop an infection and what is even very interesting in this experiment then they did the experiment again and they say okay we have a salmonellosis problem in our farm so we are going to vaccinate against salmonella and they repeat the experiment the same way here they use only the animal feed with a control feed or with high level of okra toxin they didn't have to wait 13 days they have to wait much longer but after 47 days in fact out of the animal receiving the high level of mycotoxin in the feed one developed brachiospheral infection and one compilobacter infection so basically it really indicates that if you have mycotoxin in your feed and you don't have a good sanitary status whatever the pathogen that is around the animal will catch the pathogen and will be infected by this pathogen so this is the first set of example that i show you indicating that the fact that mycotoxin decrease the immune system will increase susceptibility to infection but there is also the immune system it is also used during vaccination and it has also been demonstrated that the fact that mycotoxin decreases the immunity will have an effect and will decrease vaccine efficacy here is some example so here it's an experiment with aflatoxin on viral infection so basically there was more than 600 cheeks two months of age that received uh feed contaminated with aflatoxin and they were vaccinated and then challenged they were vaccinated to different infection and then challenged with uh this um the the pathogen and basically you can see that in the first set they were challenged with newcastle disease there was an increased antibody uh title and leading to an increased mortality in the animal that received aflatoxin exactly the same result with infectious on bronchitis there was a decreased antibody tidal and increased mortality after challenge by contrast with infectious birth bursal disease there wasn't decreased antibody title but there was no effect on mortality this is another experiment that we did in our lab with the vaccination and it was a mycoplasma vaccination again macroplasma and we when we look at the total antibody response there was absolutely no effect but when we look at the specific antibody response you clearly see that at the end of the experiment the animal that received the feed contaminated with aflatox with feminism sorry get a lower antibody response than animal receiving the control feed in this case because we are scientists we try to look to look at the mechanism involved and we see that it was because of cytokine because there was an alteration of the th1 th2 balance that is really important for antibody production and in addition we had a decrease in fossil proliferation that lead to decreased antibody production and lead to decreased vaccine efficacy i think that's quite important because breakdown in vaccinal immunity induced by mycotoxin may lead to occurrence in disease even in properly vaccinated pigs and the problem also is when you get the disease you don't you know that you have vaccinated your animal but you most of the time you don't remember what type of feed you were giving to your animals once or two months earlier when you were vaccinated your animals so if we try to conclude on this part we can see that mycotoxin can alter several aspects of the immune response i show you that t2 toxin alter the production of antibody whereas aflatoxin b1 alter the lymphocyte proliferation and this really have some consequence in term of pig health or even in animal health in general it increased susceptibility to infection i didn't show you but there are there have been experiments also showing that it did it reactivate chronic infection i show you that it decreases vaccine efficacy and there are also experiments that show that ingestion of mycotoxin may also decrease drug efficacy however in most of the case the underlying mechanism still need to be elucidated i show you in for feminism b1 we did some experiment and now we know uh what is the mechanism it it is due to cytokine production and lymphocyte proliferation but for some other case it's really not known for example the case of oprah toxin that i show you that decreases that ingestion of okra okra toxin increase susceptibility to several infection we don't know exactly the mechanism so i'm almost finished this part of my talk but before finishing i wanted also to tackle a new problem that um is today on mycotoxin and this is the problem of mycotoxin co-contamination co-contamination is a reality it's a reality for several reasons the first reason is that fungi produce several mycotoxins simultaneously because also feed may be contaminated by several fungi at the same time and because the animal diet is composed of multiple raw material for all this reason we have a co-contamination and this is for example a survey that has been done in almost 10 years ago on more than 7 000 samples and in this survey it indicates that about 20 percent get low level of mycotoxin below the limit of detection but 33 percent in this sample there was one mycotoxin but in 48 percent they detect more than one mycotoxin and so contamination by several mycotoxins is a rule not the exception and most studies have investigated the effect of mycotoxin when present separately and we really need now to investigate the effect of mycotoxin co-contamination what are the effects of disco contamination so the experimental design to study mycotoxin mixture you need to have a two-step approach first you need to do a dose effect of each mycotoxin when used individually and this is quite important this is just two example two dose response example in the one on the top you see that when you multiply the level of a contaminant by two from two to four the response is only increased by 13 but in other example in the bottom when you go from two to four you increase the response by eleven fold so you really need to first have those response analysis and then when you have this dose response analysis for each mycotoxin you can compare the predicted effect of the combination with the single effect of the combination and then you can determine if they have no interaction of if they have synergistically or antagonistically and the big problem is that many studies addressing mycotoxin interaction are very difficult to interpret because there is no dose response experiment we did some experiment with mycotoxin mixture especially on cell on human cell or on porcine cell and we see that in the vast majority of the case we observe some synergy here it was with different trichotic and dawn and three acetyl done done and 50 lasting dawn fifteen and three are sitting down don and ivana dawn and from infusion on x nivalinon and fleurs are not x and you see that at low concentration in the vast majority of the case the main type of interaction is um observed is a synergy so that really indicate that this energy need to be taken into account so um now i go to my conclusion so mycotoxin have multiple effect on animal and among them they are able to modulate the immune system and the main consequence of this is in terms of susceptibility to infectious disease and in reduced vaccine efficacy something that i didn't show you also is that mycotoxin are able to alter the intestinal barrier function then the intestine is not a barrier anymore and these have some consequence in term especially in terms of pathogen translocation across the industry what i show you is that despite of the frequent co-occurrence of different mycotoxin little is known about the interaction between this toxin and also something that is new is that there is more and more new fungal metabolites that are identified emergent mycotoxin mask mycotoxin and now we really need to address their toxicity and with that i finished my presentation and i thank you for your attention and i'm open to question i'm going to remove my screen so i can see the question isabel now we don't see you no she probably left the stage it's possible yeah margo can you help us to bring isabel back yes isabel is coming back here she is margo thank you thank you yeah margot also thank you very much for support isabel it was a fantastic presentation thank you very much uh it raises several difficult questions or at least for me and then we would like to ask you to help in answers and the first question was how do you test the immune response if you are struggling to find a good antibody titer to test antibody titers from dead vaccines are not very good so i i think the the real problem is that we don't know so i agree that sometimes it's difficult to see antibody response to vaccination and so then it's difficult to see how mycotoxin can affect the immune response because also maybe in this scheme the mycotoxin will not target the in the antibody response but they may also target the lymphocyte with proliferation so i think unfortunately there is not one response that tell you what you should do to determine the effect of mycotoxin on the on the immune response but you need to do several tests do you have any advice on what type of vaccine to test for instance in chickens or pigs maybe you have already some experience also field experience i think it's it's difficult because um in laboratory you need a vaccine where you know exactly how to measure the response to the vaccine and in the field it's much more difficult for example in laboratory why why i was using a model vaccine it's not because i prefer to use i would have preferred to use a real vaccine than a model vaccine but to use a real vaccine you need you have you need the company to give you the antigen that is making the vaccine and usually it's kind of secret so it's why i'm using a model vaccine because then i know exactly what i'm giving and it's easier to determine the response of the animal thank you exactly the next question is there is at one article mentioned that t2 reduced the intestinal colonization but enhanced this is systemic distribution of this bacteria in the animal body what would be the explanation [Music] that can be that can be different thing because it can really also the problem of the immune response is you can have a local immune response that can be very different from the systemic immune response so if you have a local and a systemic immune response that are different you can have one effect locally on the intestine and a different effect or at the systemic level okay thank you [Music] there is another question when addressing the impact of mycotoxins on the response of the animals to any xenobiotics you advise to always consider a combination of low doses of mycotoxins as often happen in practical situations what are the most relevant combinations okay the most relevant combination really depends on the on the feed that you are giving to your animal for example feminism is only present on maize it's never present on on wheat so it really depends on what you are what you are using and i think it's also depend what is your question because if the question is is it better to give a non-contaminated feed than a contaminated feed i'm not sure that you need to do an experiment but when you need then to really understand at the mechanism level what is happening then you need to have a feed that is really well characterized and you need to have control for example when you speak about mixture if you really want to understand what happened you it's better to have a co-contaminated feed and also feed contaminated with a different mycotoxin then you can see the effect you are observing to which mycotoxin is in you because you can have an effect that is only due to one mycotoxin or that is due to a mixture to the mixture and then you can observe a synergy stick effect for example okay thank you very much the next question is uh do you think that swine and poultry immune system reacts differently to mycotoxins overall i don't think so now you can have some slight difference but basically the immune response is quite similar in human and in animal and in different animal species so we would expect the same effect right um julia i missed the question that you were uh answer asking the first time i just want to avoid that i asked the double time can you please remind it was um t2 article 92 okay yeah and then um so then the next question is it's known that for monisians have very low bioavailability knowing that for monisense are almost entirely remaining in the gastrointestinal tract what would be the mode of action on the immunity that's a very good that's a very good question and that's even not only on immunity it's true that we it's estimated that the absorption of humanism is only four percent but feminism have a lot of effect not only in the immune system it induced pulmonary edema in pigs it induced leukoencephalomalacia in horses and we still don't know why it has such an important effect with such a low absorption so it's still uh to study okay thank you still not known okay the next question is um how to measure efficacy vaccination in the field probably you started to discuss this already yes yes i would say the the best way of measuring efficacy or vaccination is to have a challenge infection but you never hope that you have a challenge infection because you never that so then a surrogate marker can be uh to measure the antibody response or the length for side proliferation because sometimes the vaccination is through antibody and in some other case it's more through lymphocyte proliferation okay thank you yeah we have many questions they are popping up the next one is use of toxin binder feed additive and poultry diet what type of toxin binder is the most effective one very challenging question i'm not sure i'm the best one to respond to this question but i think then it's also depend to the mycotoxin because we know that mycotoxin have a very are very different one to each other so usually you can have separate um different way to bind the toxin but i think it was more the topic of the first uh seminar that you had that you had before me okay thank you you're right you're right it's not like the topic of this webinar but still yeah yeah it's very good many customers are asking yeah you're right but it's again it depends on the type of the challenge the type of the mycotoxins and again right okay the next questions is uh in a challenge model on swine is there a recommended cocktail of mycotoxins to see any effect on immune response [Music] what i show you in my presentation is that a lot of most mycotoxins have have an effect so to my point of view whatever the cocktail of mycotoxin you will use you will have an impact on the immune response and it's why it's very challenging because unfortunately you are not always sure to have a free mycotoxin feed and so you are sure that your animal will have a proper and the best immune response so no any recommendation any we can use any uh mycotoxin and we will definitely have negative effect on immune system rate most of the time unfortunately and can you name the number one like the most important for immune system mycotoxin it's fine for example there have been a lot of work on the effect of uh done on the immune system and aflatoxin on the immune system i think this is a mycotoxin that will have the highest effect on the the immune system then i will put feminism and last i will put okra toxin so don't don't is number one even in the immune system right and is those which are important because they have different uh toxic levels uh known for monizing socratoxins for instance if we would like to challenge swine or we expect some immune effect of hormone agents it would be the same level as dawn and ochre toxin or we would need a lower high level uh yes it's in most of the case there is a dose response the effect on the immune system is those dependent so the more mycotoxin you have the more effect on the immune system in the immune system you will have so usually it's really deleterious and the more mycotoxin you will have the weaker your immune system is going to be and then the more consequence you will have in terms of susceptibility to infection and if you would expect the same response of immune system from for instance one ppm of dawn one ppm of homology or one ppm of ocratoxin no you have in terms of potency you have mycotoxin that are more potent than other for example a feminism you need to use higher level of feminism to get an effect on the immune system compared to those so it might also be related to the poor absorption of feminism and the high absorption of done for example but it's true that the effect on the immune system will not be the same on a milligram per kilogram feed basis for each mycotoxin okay and we have next question connected with dawn uh what is the mod of action of dawn on immunity in which organ this takes place so um what is the the mode of action of don uh the mode of action of dawn is i would say there is two effects the first thing is the mod of the dawn will bind to the ribosome and this will lead what we call um ribotoxy stress this ribotoxy stress will first give a very [Music] burst of cytokine which is not always good because when there is too much cytokine this that's not very good for the organism and then it will decrease the immune system on a this is a in short term we have an increase of all cytokine and long term we have a decrease of the immune response on which organ the immune system the effect on the immune system will be very diffuse because it mainly acts on lymphocyte and there is lymphocyte all over the body so the it's more difficult to say to say if there's an organ specific for the immune system than for other pathology because the immune system is very diffused on all the organ all the body uh yeah um i have another question how do the levels of mycotoxins used in your experimental models compare with those seen in commercial situations either alone or in combination do they represent uh in the first experiment that i show you the level that we use in the lab are usually higher than the one that we see in in the field but on the other hand when i'm doing an experiment in the lab i'm doing one or two experiments per year so between two experiments there is six months where the facility is completely empty which is not the field situation the number of animals per pen is lower than in the field situation so i know that i'm using a higher level of mycotoxin but this is still a model because this is not the real situation where there is quite a lot of animal that there is a lot of animal around so i know that it's a higher level but i think on the other hand the way we have we are handling the animal are also different from a real situation but the lowest level are close to the maximum recommended level so indicating that you can have an effect with unfortunately already recommended maximum level of mycotoxin if i may add my observation when we make animal trials of course a number of animals is very crucial for getting a good statistical data or significantly different data so and of course in the practical conditions with thousands of animals probably we can get the much more visual results having even lower levels of microtoxins but in the university sometimes we are restricted to have a number of animals and for instance dealing with only seven animals we have to really bring the statistically significant data and of course we have to challenge anyone more to see the first results and see the differences exactly okay and we have another questions um what do you mean in your presentation by need to stimulate the immune response to be sure to be able to see an effect what i what i was [Music] meaning is that basically the immune system is very strong if you look at the antibody level of an animal with or without mycotoxin in the field the total antibody level will be the same it's only when the immune system needs to work when it's challenged that you see that mycotoxin will have an effect for example if you look at just the anti i'm going to take the example of antibody again but if you look at antibody level total antibody level on animal you will need very very high level of mycotoxin to see an effect but if you vaccinate an animal and you look at the antibody level for this vaccination here you have stimulate the immune system with a vaccination here you will need a much lower level of mycotoxin to see an effect it's what i'm it's what i said that i need to stimulate the immune system on the steady state level the immune system when the immune system doesn't have to work you need very high level of mycotoxin to change it but when the immune system is acting to fight an infection to respond during vaccination here the level of mycotoxin that is needed to alter the immune response is much much slower okay thank you and the same as you showed that when after vaccination you had uh challenged by viruses from the uh destination also it was like stimulation of immune response yes and you was able you were able to see the negative effect of mycotoxin right it's like again stimulation of immune system exactly and the bacteria and the coccidia can be also the same yes okay challenging immune system we still have a couple of questions and almost finished what do you consider as low doses of mycotoxins in a combination approach what i consider as a low dose of mycotoxin is those that are close to the regulatory limit because i think the regulatory limit are very important because this is the maximum limit that are tolerated in in the feed so if you have an effect with those dose it mean that they are not protective as they should and the be is based on the idea that if you follow the recommendation you wouldn't have any effect because it's why recommendations are made so if you have an effect with this level i consider them as low-dose we have participants and from different countries and different countries apparently have uh different levels which are accepted for the feed does it mean that animals uh in one country are more resistant to like examples but i feel that the field of legislator who made that legislation i don't know why in some countries they estimate that they can tolerate this level of mycotoxin and another country they generate lower level that's not a scientific problem that more a risk assessment problem i think the eu in that case is quite advanced i heard that also efsa is considering now to reduce the physiologically safe or accepted levels for the feed that there is a discussion and i hope that you probably can give some highlights on that as well to us or is it still a secret no i i think it's true that in term of regulation europe is quite strict even more strict than most of the the other country so um efsa didn't did not decide to reduce the level or not but it's true that from time to time the european um [Music] commission asked efsa to re-evaluate the mycotoxin but also other contaminants with new data and sometimes new data show that we have level you have effect of lower level and then if you show that then you reduce the the tolerable level but this is this is quite a this is quite normal the more data you have the more uh accurate uh effect you can have and the more accuracy you can see at these dose you have an effect or these dose you have no effect okay thank you very much and we have a couple of questions left so one of them is uh is there an effect of age on the response of the immune system to mycotoxins what are the most critical situations youngs breeders or else usually but in in general not just for mycotoxin but young animals are very susceptible to infection because young animal usually their immune system is not completely mature and for example to have a completely mature immunity you would need to win pigs around i think six weeks of age which nobody does so uh usually younger animals are more susceptible because their immune system is not completely immature and you also have a problem with very old but this small for human because usually you don't keep farm animals that very old what about breeders for example we are keeping them little even breeders are mothers older as elderly people okay can they give their immunity to their offspring it depends sometimes yes some time no it's either through the placenta or through the colostrum so that's uh but this is this is true that people try to give immunity from mother to offspring which will be very good for the offspring at the when they are born meaning that if the animals like breeders has mycotoxins in their feet they have some negative effect on immunity and this immunity will go through the cheeks for progeny that have been demonstrated for example for aflatoxin if it has been there are some papers showing that when mother were filled with aflatoxin contaminated feed there was a decrease in unity on the offspring but it really taken on mycotoxin i think for john it has never been demonstrated neither uh nor for feminism but it has been demonstrated for aflatoxin but does it mean that it's it's not true oh it's just not be demonstrated you it's not been demonstrated so uh it's a potential uh top potential for the new research okay last question and then we will wrap up because i have a message also to participants the last question is do you think that the combination of those below limits might be significant you mentioned synergistic effect i think this is a real real question and this is a big topic to know whether and it has been demonstrated for some mycotoxin not always for the immune for the immune response but that when you have all the mycotoxin at the maximum limit you still have some effect so it really indicates that there is an interaction between the different toxin okay thank you very much uh last question just about the synergy which synergy is well do you know um how can we you know do we have any issues or any possibility to calculate the synergism of different microtoxins i know that there is some creation some methods but we don't use them in the practice maybe no that's that's very complicated because the problem of the synergy is really it's only it's infancy we don't know much and so far all the regulatory limit or everything they don't consider synergy they consider additive effect and even if you look for example from efsa most of the time now it's not regulated mycotoxin by mycotoxin but family of mycotoxin for example it's aflatoxin b1 b2 and m1 that are regulated together feminism b1 b2 now it's done and modified form they are in on unmodified form so now we have a group group tdi for all this mycotoxin but most of the time it's done we you consider that it's with additive effect we don't consider synergy because it's too complicated and if i want to make the story even more complicated giulia is that we are not only exposed to mycotoxin and for example in our lab we find interaction between mycotoxin and heavy metal that's interesting for example so we are not just exposed to mycotoxin and cadmium at really high levels or just elevated levels at uh so now we go uh at levels that are possible in the in the in the field really oh yes that's something that's even more that's even more complicated for the story wow yeah exactly yeah okay okay let's wrap up uh i would like to thank you isabel very much for precious information that she shared with us and also answering uh the challenging questions and giving a really a very valuable information um so we will wait uh for participants for another session but i also got some messages that some people were joining late today and they would like to get a record of the presentation i would like to say that there will be a replay of this session next tuesday at three o'clock paris brussels time so and those people who were missing some talk of isabel can also see the records next tuesday you can register for that [Music] so thank you very much and i wish you a very nice day evening and hopefully to see you soon back thanks a lot isabel thank you okay have a good day