everybody and thank you for your attendance to this webinar it's an honorary pleasure for me to speak about this topic in this talk i'll speak about binding and mycotoxin absorption and i will give you the options that we can have in these fields of science toxins are secondary metabolites of some microscopic fungi they are not associated with growth or primary metabolism their biological meaning is not completely clear they are produced in optimal conditions of temperature and humidity and they are stable to the physical chemical and biological processes of food processing the problem is that mycotoxins are toxic for humans and animals and they are able to induce microtoxicosis in this slide you can see the chemistry of the most important mycotoxins that you can find in foodstuff they are not a unique chemical class they have a relatively low molecular weight a moderate or high polarity low volatility and high stability each mycotoxin is produced by one or more fungal species and in some cases a fungal species can produce more than one mycotoxin here an overview about the presence of mycotoxins in foodstuffs as you can see the most of mycotoxins are present in corn corn by product in cereals there is a toxin m1 that is a big problem in milk and derivatives and the problem here usually is that there will be an occurrence of mycotoxins in a lot of foodstuffs there are different decontamination detoxification strategies that having they have to have in common some futures they have to inactivate destroy or remove the mycotoxin avoid the production of toxic residues in the final products an important point is that they have to retain the nutritive value and food feed acceptability of the products preserve technological properties and be economical simple and not time consuming among these strategies you can find physical strategies such as removal of fines clinics gamma radiation and so on the use of different chemical compounds to reduce microtoxin contamination biological strategies such as microbial degradation and feed food processing so thermal and mechanical treatments applied to raw materials the problem is that these strategies show different drawbacks for example physical treatment are usually expensive with uncertain results often connected with high feed food losses so with limited practical application concerning chemical treatment they are expensive to usually change in palatability and the nutritive value and the important point that toxic by-products are possible so no practical application an alternative to these strategies there are feed additives affiliatives european regulations in 2009 established a new functional group of feed additives in the category of technological artists these feed additives have to suppress reduce absorption promote the excretion and modify the mode of action of mycotoxins anyway no increase of the existing max guidance levels of mycotoxin feed are done anyway to uh put in the in the market i feed additives uh it needs an authorization procedure so uh the applicant need to prepare and submit a dossier according to the fsa guidelines responsible for the valuation of the data and can provide a positive opinion after this positive opinion this product the feed additive can be released in the market in this slide you can see the three feed additives regulated in europe as mycotoxin detoxifying agents the first one is the bentonite uh using uh used and feed additive for ruminants pole trim picks as afrotoxin binder the second one is a microorganism to uh biotransform trichotisms for pigs and all agent species and the third one is a group of regulations concerning an enzyme for degradative femoris in contaminated field of course other feed active can be registered in in this regulation if the applicant wants to invest some extra cost to register his product for saying the requirements of the assessment of the microtoxin detoxifying agent we need to characterize the additive uh by some instrumental methods we need to assess the safety of the the feed additive in particular for mycotoxin binder we need to assess that no adverse effect on digestion and absorption of nutrients uh happened and of course we need to evaluate the efficacy so to specify uh for which microtoxins additive will exert its function and target species and to declare and demonstrate the mode of action of the additive if is an absorption for example or a biotech transformation a thickest assessment can be done by in vitro study a very potential tool to screen a different microtoxin the toxic anagens and to study the mode of action of the additive but it does not mimic the conditions of the digestive tract so efsa in europe required individual studies to demonstrate the efficacy under practical conditions by short-term studies in target species the analysis of mycotoxins and metabolites can be done using the biomarker approach to evaluating microtoxin excretions in physics urine or concentration in blood plasma serum or in tissues or animal products concerning the methods to assess the efficacy of the product we can have in vitro study and inside this study we found the absorption trials using a single concentration of microtoxins adsorption trials using different concentration of mycotoxins or different adsorbent dosage of the product so this is the famous methods of the isotherms we can also study uh the desorption processes to understand the strength of binding between our product and microtoxins and to understand the more specific physical chemical properties of these processes we can still we can study also the thermodynamic of that solution we can use the for a fixed assessment also gas intestinal models static or dynamics and in the end in vivo study you're studying no specific parameters such as the state of the health of the animal weight of the animal and so on or specific parameters the biomarkers approach what we can say about the literal vivo comparison both of its of these triads show advantages and drawbacks concerning in vitro advantages foreign welcome to everybody organic binders such as east cell wall agriculture by products or other organics polymers inorganics binders the most famous clays and synthetics binder in particular modified minerals such as organoclay activated clay or mixture between organic and inorganic binder more than hundred commercial products are available on the market most of them claim the ability to absorb several toxins but simultaneous multi-toxin absorption has been poorly addressed okay we can start with an overview about organic binders i want to show three examples of this kind of binders starting from each cell wall these in vitro studies show how it's a wall used at two percent of weight per volume dosage absorbed more than eighty-five percent of geraldine on ocratoxin a and the fumonism b1 at ph3 and seven used to simulate the gastrointestinal tract of the animal whereas afratoxin d1 absorption was lower than other microtoxins how we can explain the uh absorption mechanism on this kind of binder there are different papers in literature that explain this topic i selected two interesting papers in which the authors take into account the complex structure of each cell wall that is characterized by polymers of mannose monoproteins beta declines and other organic compounds they found how beta-b glucans are the east component responsible for the complexation of the animal weak hydrogen and bodenville's bonds are involved in this chemical complex formation the authors extend these these results also to other kind of mycotoxins in the right part of these strides there is a work of yaniculocetal in which they found a correlation between the amount of beta-glucans and cell walls and complex forming efficacy increasing the amount of beta glucans in necessary walls you can find an increase in the amount of geraldine absorbed by yeast cell walls this product was also tested in vivo usually using uh 24 sexually mature girls that were fed with the three diets for 42 days first diet was the control second diet was the naturally contaminated diet with geraldine and the third diet was the naturally contaminated diet supplemented by each cell wall product at 0.2 percent of dosage during samples 24-hour post-administration were collected and the the amount of drama and metabolites were analyzed using the biomarker approach this work is in submission in a scientific journal what about results as you can see from this box plot this graph at day 42 uh the group supplemented the group of piglets sacrament supplemented with the east cell wall showed 68 percent of zeralandon and its metabolite reduction comparing to the group without a cell wall product this reduction was a statistically significant and this means that this product was able to reduce the fixed urine excretion of the ralenon and metabolites okay moving on the second examples uh i want to speak about agriculture by product in particular about great polymer promise this product was obtained uh inside the big european project microread seventh framework program and we found in some with some in vitro studies that using this product at two percent weight per volume over over dosage they promise absorbed more than ninety percent of afrotoxin one zeralinone okra toxinate despite two ph values used whereas the human gb1 assumption was lower than other mycotoxins same product was also in the same project the european project was tested in vivo in order to assess uh the ability to of this product to reduce mycotoxin bioavailability in piglets as you can see from this graph gryphoma is reduced significantly urinary mycotoxin biomarker of afrotoxin b1 of 67 percent reduction and zeroland 69 whereas reductions statistically not significant were observed for humanism one dioxin valine and ochre toxin so these are very interesting results for details about this product about in vitro and individuals can be found in these two recent papers published in the journal of agriculture and food chemistry considering the third examples of organic binders i want to speak about durian pill and in particular about its modification this work was done in collaboration between institute of sciences of food production and tamasa university in thailand why in thailand because this is a typical fruit in thailand its waste was more than 200 tons per year and it caused environmental and social problems in diet so we tested this product as such and we performed also an acid hydrolysis on this product to obtain a modification of this product called acid treated durian peel after that we evaluated the multi-microtoxin absorption activity what about results in this graph you can see that the acidity treated durian pill red bars showed the high absorption values towards athatoxin b1 xeralenone or cartoxinate and the human db1 donor absorption was negligible and the ph affected occatoxin a and fibonacci one absorption anyway the modification of this product of natural durian pill increases the microtoxin's absorption this acid pretty durian pill was also tested with the gastrointestinal static model following the minecraft protocol that can be found in the paper published in 2014 and we studied the capacity of this product to reduce the bioaccessibility in the gastric and intestinal phases of these four mycotoxins as you can see from the results acid treatment pill reduced the bioaccessibility of mycotoxins in uh gastrica in gastric phase uh and in particular aflatoxin one and zeralinone showed a reduction in the by accessibility of also in the intestinal phase also for this product for more details uh you can read this interesting paper published in 2020 concerning the effectiveness of durian pill as a multi-microtoxin absorbent of course this is not an exhaustive list about organic binders and about agriculture by products and other information about comparison of different kind of agricultural metal products to be used for microtoxins absorption can be found in this other work published in 2019 comparative efficacy of agriculture byproducts in sequestering mycotoxins okay moving on the second category of binders i want to speak about inorganic binders in particular about clays clays belonging to the philosophicate class and to the spectac family because these are the the most uh an organic binder studied in literature concerning the mycotoxins absorption these plays are characterized by different sheets of tetradic octahedric and tetraelic structure t-o-t sheets and this nectar family can be divided into the octahedrals magnetite and three octagonals mechanic inside the deoctohedral necktie we found montmorillonite very famous minerals to be used for microtoxin absorption and inside the trioctyler spectite group we can have for example saponite or hectorite among these magnetites the octahedral or theocratic we can find bentonites bentonites are clay minerals belonging to the phyllosilicate class and to the spectate group i want to speak about bent light in particular because in europe bentonite is authorized as a feed additive particularly as a substance for reduction of the contamination of bioflotoxins for ruminants poultry and pigs ventrites received the claim 1m558 to be used for this issue but to be obtained disclaimer uh regulations say that bentonite has to show more than 70 percent in smackdown percentage less than 10 percent in other minerals uh contaminants and this is nectar as in the form has to be in the form of the octahedral montmorillonite and above all after a specific absorption essay described in the regulation aflatoxin one binary capacity uh has to be more than ninety percent or we can explain the mechanism of that suction between spectites and mycotoxins in particular autotoxin b1 there are two uh most accepted suggested mechanism for this kind of process one is the electron donor acceptor and the other one is potential collision electron donor acceptor in this mechanism uh outers explain how spectites are negatively charged due to isomorphic substitution in this structure and so they can attract positively charged ions to balance discharge this positive recharge can be the carbons in the carbonyl system of a flatoxin b1 that are partially positive moreover aflatoxin b1 is planar so can go inside the interlayer space between one sheet and another one in the philosophy it can be absorbed so in terminal region from this mechanism interlaminal region is the primary site of the binding of afrotoxin b1 instead external surfaces accounting for only minor absorptions and the external surfaces can explain also the absorption of other kind of necrotoxins that present a structure different from afrotoxin b1 potential calculation explains how mycotoxins in particular afrotoxin can be called by some cations present in the interior space of the smactites or by some various edge site metals anyway not all doctrinal neckties are the same in our inner city study uh performed in a big european project microkey project we found our zmectites uh photoxyl1 absorption from zmectites can be affected by geological origin of these minerals we divided the spectites in two groups in a sedimentary group and hydrothermal group concerning taking the count of daily geological origin and we performed absorption ice attempts on these products as you can see from these box plots we found that sedimentary spectres show significantly higher values for maximum absorption capacity and absorption affinity than hydrothermals magnetites so geological origin can be an important topic to to see to use this product as a toxin b1 absorption for more details concerning the role of geological origin and their physical chemical properties on a fatoxin absorption you can be uh observed you can be read this paper published in 2019 on the applied clay science journal natural peptides are able also in uh to absorb humanity one but as you can see from the graph this absorption is affected by ph in particular humanism one can be absorbed at ph3 and with the less extent at ph 5 but assumption at ph 7 is negligible you can see an um an example of uh a montmorilloni uh montmorillonite clay used in vivo in rats the outdoors uh micheletta uh explain uh how this product this commercial product is able to reduce aflatoxin one and humanism b1 biomarkers in rats exposed to single and co-exposures of toxins they study uh pharmacokinetic uh exclusion of fratoxin m1 in urine and humanism b1 in urine of rats and they found a significantly reduction in bioavailability of afrotoxin b1 if you wanted a new one in this animal model so they conclude that this product may be used to reduce toxins levels below the thresholds for affecting adverse biological effects okay it is like the uh i want to focus the attention about the uh direct hydraulic group and the chocolate group of smactite even if the regulation uh explained how smacktite has to be in form of your headless neck or the octahedral structure in our experience we found how following the european regulation the absorption essay the screen described in regulation also other kind of smactite in particular trioctyldrazonectite can be fulfill the regulation and can be reached at the 90 percent of a fretoxyb1 binding capacity so since that this kind of group of minerals can be properly excluded by the european regulations here you can see an example of the use of a trioctyledrasmictite in vivo this is a big european italian project farming project and in this project we studied the the capacity of these three octahedrons neckties to reduce aflatoxin and one amount in cows in bulk milk cows exposed to afrotoxin b1 as you can see from this graph the group of cows [Music] treated with the trioctyledrasmic diet showed a significantly reduction in the aflatoxin m1 concentration and the flux toxin 1 total excretion in bulk ink so this product seems to be a promising product promising minerals to be used in animals okay moving on the last part of uh binders the third categories i want to speak about synthetic bentonite based material and i want to spend a few words uh on organoclay that are very famous uh synthetic uh materials organoclades are characterized are a mixture between natural smactite natural bentonite that are treated with an quaternary alkyl ammonium ions these categories kill ammonium ions can go inside the interior spaces of smactites and can be characterized and they can be obtained a new organophilic compound to be used for different issues such as microtoxin assumption as you can see these three examples of organoclay you found a very good mycotoxin absorption values for aflatoxin b1 zeralinone ocartoxinae fumonismi1 and typotoxins despite two ph values used dioxin valeno assumption was negligible also for this product concerning a carbon activated carbon or mixture between ventricles and carbon these are very good in vitro products they can they are able to bind almost hundred percent of all mycotoxins uh and for activated carbon also done is absorbed on these minerals these two categories of products show but show are very good as in vitro binders but they show some problems organocrates are not suitable for fib ingredients due to toxicity of the interlayer categories and carbons is a specific sequester so this reduces possibilities for its practical application okay another interesting product that can be used as a multi-toxic solvent is activated clay in this in the european uh project uh big european project microkey we obtained this new product selecting a robin light and activate activated and functionalized this row bentonite with an acid the window an organic modifier in order in order to obtain a synthetic new materials that was done in vitro and in vivo test test it concerning in vitro speakers assessment using a very low absorbent dosage of this product one kiloton as you can see from the red bars activated bentonite showed a higher microtoxin absorption values comparing with the native bentonite these results were confirmed also by absorption isotherms studies here you can see an example of an absorption isotope using the language mathematical model and we found that for these activated clay maximum absorption capacity values determined for aflatoxin b1 humanism 1 were higher than values found for deralon and ochre toxinate another interesting studies on this product was the desorption studies the social studies consist in an absorption at ph3 and then in adsorption at ph 7. in this study we performed also adsorption in a strong organic solvent to understand the strength of binding between activated clay and microtoxins as you can see from the red and green bars a change of medium ph from ac to neutral did not release the absorbed toxins and in particular afrotoxin one and humanism one state kept bound on the minerals also after an extraction with metal this to underline the good binding between this activated clay and microtoxins this product was also tested in vivo using two different animal models rats models and piglets models and using two different experimental design rats were divided into two groups one treated with activated clay and the other one untreated with activated clay a single integrative bolus administration of each mycotoxins was was given to the rats and urine samples at different endpoints were collected to perform a toxiokinetic studies concerning piglets we by a single intragastic bonus administration of a mycotoxin cocktail of aphatoxin b1 nocatoxin in humanism one and zero element and dividing again piglets in two groups uh treated and untreated with deactivated clay in this case blood samples at different time points were collected and also in this case a toxicokinetic studies were performed the analysis of mycotoxins were well performed using the biomarker approach okay what about the results concerning this results results we studied uh what's happened are two important uh at least two important toxicokinetic parameters such as aoc area under the curve and maximum urine or blood concentration in of microtoxins concerning rats we found in the ioc a statistical statistically significant reduction of the aussie curve comparing control entity group for aflatoxin m1 or catoxin a and human db1 we didn't find any lc reduction for the end metabolites but we found a statistically significant reduction in the cmax values an important another important toxicogenetic parameter so you can see that this product activates clay is able to reduce bioavailability of mycotoxins in rats concerning piglets trials was found a statistically significant reduction in alpha toxin b1 for aflatoxin b1 endocatoxin a values in the oc values comparing control and treated pulp and a trend reduction was found also for humanity one and four zeraland on gluconide that in in uh blood uh in piglet's blood is the biomarker of geraldine for their allele no seed reduction was found but was found the trend reduction for the c-max values how we can put together these uh these trials in vitro trials and in vivo tires of this activated clay we can say i would to say that first of all before to try in vivo product we need to we need to have a very good mycotoxin binders in vitro as you can see this product was able to uh to absorb more than 82 percent of mycotoxins of this mycotoxin tested and concerning in vivo despite to the different experimental design between rats and piglets models results were quite similar for these four mycotoxins and i would say that it's rats models can be a good model to test a prior product in vivo before to go to other more complex and with difficult management animal models such as piglets or other kind of animals okay concluding my talk i want to summarize these results and you can see that majority of bad sorbents uh activated carbons are ineffective to add sodium oxide volume each civil based products absorb preferably xeralinone uh agriculture by products such as great formation or durian pre-modified are quite effective for afrotoxin one zeralinone endocrine natural clay minerals absorb preferably aflatoxin b1 at ph3 and seven endocrine toxin a and humanity mu1 at ph3 organoclase can act as multi-toxin and solvent but they are toxic so this reduce possibility to uh or for application in vivo and activated clay can be a promising potential multi-toxin adsorbent to be used in vitro in vivo for different animal species so i want to thank you for your kind attention if you have some questions sorry talking to myself here so um we have uh the question area on the side you can open that uh question box just type it in sent to us it can be in spanish or portuguese or english we can translate and ask the questions the answers though will be in english but we can send uh on an email as a follow-up in spanish and portuguese dancers as well so i will give a couple minutes for questions as i start to receive them here you will have another 15-20 minutes for that okay let me ask the first uh question to dr olga verkieva how to compare the different microtoxin binders in the market uh it's a very relevant question a lot of customers have issues to compare different products and the most available tool is to compare in vitro efficacy but i would not really recommend to use it as a only tool to compare products the best of course if you can test your product in your animals if you cannot do this i would advise to request as much data research data from the producer of the product as possible and please focus on microtoxins which are relevant for your situation if you have dioxin valeno issue you can't use data on aflatoxin studies or if you have pigs you cannot always rely on data that you get in poultry especially in specific microtoxins like zeralinone which has different metabolic pathways and different species in vitro binding data could mislead you and you could see it on one of the latest slides of dr vito that there was a binder binding azira lenon at 99 percent but when it was tested in vivo it was not it was not effective at all so it's always important to have in vivo proof okay thank you i have a question here to dr julia um what about phytogenics to recover liver or other tissues damaged by microtoxins yes that's very good questions they are really important but first of all we need to eliminate their mycotoxins from their from the body as much as possible and after then we can start taking care of the liver and all other organisms or organs we can support during these mycotoxic causes because even in good cases we cannot uh absorb 100 of mycotoxins for example even when we speak about aflatoxin b1 uh we know that in vitro studies absorption is more than 90 percent even 100 but we will never see the same in vitro in vivo meaning that maximum 80 percent would be able uh to bentonite would be able to absorb for example aflatoxin b1 and the rest will go to the liver and in that case we will definitely need something to support liver to to help liver so phytogenic in that case would be help helping a lot thank you we will have another session one of the five we have for this set of webinars which will be also devoted to the organ support and i think this question will be even more answered uh during this session in april i believe should be in the end of april next question uh dr oga is there a standardized efsa absolute test for my toxin binding which is accepted in other jurisdictions uh this is available uh only for aflatoxin and this is standardized for uh binding with aflatoxin by um 200 grams of bentonite per ton of feed equivalent at ph um five and this is available in the european legislation for bentonite with number one zero six zero from 2000 uh 13 i think so this method to describe in the law okay thank you next question uh can be to either of you can you please talk about different methods of activation i think it means about uh the clay activation olga would you start um yes probably i will do there are many different activation uh processes and the most known is the process of activation of natural clays with cations that for instance bentonites in nature they are existing mainly as a calcium bentonite but calcium bentonite is not very good in binding capacity for aflatoxins for instance and producers of bentonite in order to approach the feed industry or microtoxin binding producing binder producer companies they are activating calcium bentonite with sodium salts and sodium has more potential it charges the layers of clays with more negative charge and it's more potential for aflatoxin binding another way of activation this is also extreme i would say it's activation with organic uh molecules like quarter nariamo ammonium or they also called quats and in some markets of latin america this organoclase are also existing they are the same principle like sodium cations but they have much stronger positive charge and they are bigger molecules and they are resulting in very very high absorption and they can absorb a lot of molecules of mycotoxins but they are non-selective they can also do vitamins minerals but the problem with this organoclase they are toxic because quarternary ammonium is a detergent it's used for washing powder or some disinfectants which are not allowed for using feed there are some patterns available uh for activation of clays with for instance collinchlorite which is organic molecule but it's more gentle it's not toxic to animals can be used in feed and many other molecules could be potentially used for the activation organic or inorganic molecules okay thank you next question uh real minerals in gastrointestinal tract interact with microtoxin binders bentonite activate charcoals and others dr julia um yes that's a very good question and it's very often asked um of course any clays any compounds will interact with nutrients with my with vitamins vitamins and minerals and even bind to some extent but in the levels the most of products used like we used bentonites uh the absorption of the vitamins and minerals will be not significant meaning that it will not uh interfere with the main function and speaking about the activated uh carbon it's uh it should be used in in high levels and it's like more uh binding capacity is more potent on this compound and meaning that the binding capacity of against vitamins and minerals can be higher and of course most of the experiments with the binding of vitamin minerals is done in vitro and you can find very nice reviews even from the recent papers showing that the clays can bind very good vitamins and minerals but again you need to to understand that we can it's difficult to compare results in vitro with real real results in vivo uh even it's the same ph level and so on the conditions will be completely different with different uh many other parameters with influence on these these things and for example we have nice results showing that mycotoxin negatively effect antioxidant system and we measured level of vitamins in blood of piglets for example and seeing that the level of vitamins uh was significantly decreased when the mycotoxins were in feed and then when we used product microtubules managing product level of vitamins was significantly higher and even higher in control meaning that there is if even there is some adsorption but the level of that absorption is not significant and it's not interfering with the old function this antioxidant system in this case so well if you'd like to comment more uh the binding capacity or binding affinity towards different vitamins also depends a lot and the most sensitive would be probably oil soluble vitamins and some vitamins of b group maybe b1 b2 but not all of them but indeed the in vivo proof showing that there is no interaction uh levels in blood for instance like julia was mentioning this is a very good confirmation that product is friendly to to premixes to essential nutrients but of course products which have already in vitro binding capacity very strong and they can bind non-selectively mycotoxins that there will be already risk that they can bind also some vitamins and minerals um so it's a balance let's say you can't say that product does not bind anything that probably it will be not effective against microtoxins and if it binds everything that it will be probably binding also essential things yeah agree it's all about balance thank you next question here is uh inclusion of my toxin binders can you talk about inclusion of my toxin binders and if there's a limit of how much clay you can use on a diet uh i will take this question it depends again on clay and depends also on the legislation in the country like for instance for the european community for bentonite there is a certain limit like not more than 20 kilo per ton of feed but 20 kilo it's a lot so normally the binders are used at maximum 5 kilo per ton of feet for other clays as far as i know there are no limitations but i am not i don't know by heart how fda limits the use of clay but i would say if you use uh binders or clays for whatever reason maybe for not for mycotoxin binding if you use it below 5 kilos i would say there will be no worry and if you have a worry it will be maybe around the nutrient binding in in small levels it would be uh no problem even i recently came across the article from medicine human medicine and they are using cpo light by the way cpu light is also part of our products in most countries of the world the pure light is used in human medicine as a carrier of some nutrients for slow release so even it's beneficial in fact interesting okay thank you um nice question what about t2 they act by contact what's the most important mode of action of this product with d2 julia do you want to answer okay yeah i will start and all doesn't comment yes sure for for t2 especially in portrait we can we can very often see these oral regions and meaning that it's so difficult to to act in this case when it starts acting by contact already already in oral for taking um leading to these oral lesions but again you need to make everything to in order to to get the mycotoxin from the body and then support support the digestive tract and support the epithelium as much as before as possible to decrease this negative effect and we have even excellent results with t2 toxin when we saw this um a very like severe case in the field in layers and then we used our product and in two weeks the oral lesions were significantly decreased meaning that even if we cannot remove from the mouse from the from the beginning of t2 we can even uh with other other means we can decrease this negative effect uh yeah it's important to know that binding is happening only when feed is mixed with gastric juices so in the liquid uh liquid surroundings so in the mouth there is already the first contact and it's uh quite naive to believe that we can bind or any product combine t2 already in the mouth in the feed which is mixed with saliva but there are some other modes of actions for instance some products contain ingredients which can be also circulating in the blood and then they increase the recovery of epithelium also in the mouth and as julia mentioned already we have field evidence we can't say that we can really protect 100 percent from mouth lesions but we can reduce notably reduce the appearance and severity of mouth lesions caused by t2 toxin okay thank you next question uh the speaker mentioned about 30 to 40 percent absorption of ones in b1 and cr lenon is that enough binding and how can i evaluate commercial toxin binders efficacy in chicken and swine feed i will take this question uh probably um i don't remember exactly when it was uh the slide for in vivo or in vitro i believe that was in vivo uh in vitro yeah but even if it's in vivo and or in vitro doesn't matter we can't expect hundred percent protection uh for any mycotoxin even aflatoxin which is the easiest to bind in some animal species we can bind up to eighty percent or uh give a protection up to 80 percent in some species uh it's not possible like in swine because it's very fast go into the bloodstream same for crayon it's a molecule which is absorbed already in the smaller intestine and the upper parts it will some xerolino will escape from the binding and will go to the blood so of course when you have high levels of xeralinone you can't believe that you have a product or you can find the product will protect your animal hundred percent so uh what i would advise best to choose a product with the proven efficacy in vivo because in vitro you also could see on the slide that 99 in vitro was resulting nothing almost nothing in vivo in pigs and rats in reds was something like 12 percent reduction in pigs was zero um if you found such product uh you play with the dosage the higher the dosage the better would be the protection if you find the effective product showing you at least um 50 of uh reduction in bioavailability and my personal advice uh nowadays that there are some bolus or toxic kinetic studies are available for testing products in in vivo so you give the bolus to the pigo chicken with mycotoxin and with the products and you see the dynamic of mycotoxin and metabolites in blood if the binder is able to bind microtoxin you will see the blood levels lower than in the control so vito was showing this type of trial and that would be interesting for you for assessment of mycotoxin binding product it will be more expensive than in vitro data but it's much less expensive than the really performance trial through several weeks because this trial takes a couple of days only okay and it's something uh we we had this the bolus trials oligo mentioned in piglets in dawn studying their pharmacokinetic so it was very interesting to see and to to see how how fast is going to uh to blood and how fast it's neutralized by the product so if possible yes it's this this kind of of trials can easily showing how it's how it's going there are several facilities are available in the world and the methods are mainly based on the publication of dr matthews matteos de frese from gent university he published his paper in 2012 and his method was applied in ghent university and you can test it here or a producer of the additive can test it here also indra toulouse isabella's wealth can do this and uh josephine aventagiato is a co-worker or partner of dr vito who was presenting today they are also doing these models thank you very much okay so we are about a minute uh to finish um we unfortunately have no more time for questions and answers but everybody that sends the question we'll try to do our best to send the answers back on an email and a follow-up um margot is just sharing a link to uh to fill up um to fill up a form just uh just to let us know how everything went for you in terms of this webinar if it's interested and of course to receive information on the next uh series we will have and i will give a couple minutes as well to mattel to say a few words to the spanish speakers so thank you very much everybody here in north america and south america and see you next time [Music] [Music] [Music] see you next time thank you thank you bye-bye