USE OF A FUMONISIN B1 AND ZEARALENONE ADSORBENT IN ANIMAL FEED

Abstract

The present invention relates to the use of an animal feed additive that reduces the toxic effects of mycotoxins, especially fumonisin B1 and zearalenone. This additive is a mycotoxin adsorbent consisting of an organically modified phyllosilicate with 71% substitution of its cation exchange capacity with the organic compound dioctadecyldimethylammonium chloride.

Description

USE OF A FUMONISIN B1 AND ZEARALENNON A ADSORBENT IN BALANCED ANIMAL FEED. TECHNICAL FIELD OF THE INVENTION The present invention relates to the use of a mycotoxin adsorbent in animal feed to prevent the toxic effects of mycotoxins, with particular emphasis on fumonisin B1 and zearalenone. This invention also relates to the use of a premixture that includes a mycotoxin adsorbent for preparing animal feed. BACKGROUND Mycotoxins are low molecular weight chemical compounds produced by fungi that have pathological effects on both humans and animals. Hundreds of mycotoxins are produced by various fungi that contaminate grains and food, whether in the field or in storage silos. In cultivated fields, the fungus that commonly affects grains is Fusarium sp., which produces the mycotoxins zearalenone, fumonisins, and trichothecenes, among others. The fungus Aspergillus sp. develops primarily during storage and produces the well-known aflatoxins, but it can also produce sterigmatocystin, cyclopiazonic acid, and ochratoxin A, depending on the species. The degree of health damage caused by mycotoxins depends on the specific mycotoxin involved, the level of food contamination, and the time elapsed since the food was consumed.Mycotoxins such as aflatoxin B1, ochratoxin A, fumonisin B1, and trichothecenes can affect specific organs, such as the liver and kidneys, disrupting metabolic processes and producing adverse conditions that lead to effects such as a pale, enlarged, and friable liver, kidney inflammation, oral lesions, decreased immune response, malabsorption of nutrients, reduced growth, decreased pigmentation, and so on. Specifically, fumonisin B1 affects the respiratory system of pigs, causing pulmonary edema, and in horses, it produces leukoencephalomalacia, both of which are highly serious conditions. Other mycotoxins, such as zearalenone and its derivatives, exhibit estrogenic activity (Diekman and Green 1992).Although zearalenone and its derivatives have low toxicity, meaning their ingestion does not cause severe harm, their estrogenic and anabolic effects cause reproductive problems in all animal species, with pigs being the most affected. In pig farming, the presence of grain contaminated with zearalenone is a problem with very severe economic repercussions due to its impact on reproduction, as it severely affects the reproductive system. Currently, one of the solutions proposed in the livestock industry for controlling mycotoxicosis is the use of mycotoxin adsorbents. These adsorbents are used as feed additives and work by trapping or adsorbing mycotoxins in the animal's gastrointestinal tract after it consumes contaminated feed. These adsorbents primarily prevent the mycotoxins from being absorbed by the animal, entering its circulatory system, and consequently causing their harmful effects. For several years, the use of aluminosilicates, clays, zeolites and even organoaluminosilicates as mycotoxin adsorbents has been known (Phillips et al 1988, Kubena et al 1990). In the prior art, we find several documents that refer to mycotoxin adsorbents. For example, document DE3810004 (1989) discloses the use of bentonite for mycotoxin binding in humans and animals. This document reveals that bentonite is effective in binding zearalenone, some trichothecenes (such as deoxynivalenol and T-2 toxin), ochratoxin, and PR toxin. However, among the most widely used mycotoxin adsorbents in the livestock sector are organically modified aluminosilicates with a quaternary ammonium salt or even a secondary amine.Thus, for example, WO 00 / 41806 (2000) discloses mycotoxin adsorbents comprising an organically modified laminated silicate comprising a quaternary onium compound, wherein said onium compound includes at least one C10-C22 alkyl group and an aromatic substituent, and wherein 2 to 30% of the exchangeable cations of the laminated silicate are exchanged with quaternary onium compounds. Among the quaternary onium compounds included in this document is benzalkonium chloride, which is a well-known biocide or disinfectant. > tu r\ c N. ac -ja -t; K a For its part, the document SL Lemke, PG Grant and TD Phillips Adsorption of Zearalenone by Organophilic Montmorillonite Clay J Agrie. Food Chem. (1998), pp. 3789-3796, describes an organically modified (organophilic) montmorillonite clay capable of adsorbing zearalenone. This paper shows that organic modification of an aluminosilicate with a quaternary ammonium compound increases the zearalenone adsorption capacity compared to the unmodified aluminosilicate. It also shows that the highest in vitro zearalenone adsorption values ​​occur when the clay is modified with cetylpyridinium and hexadecyltrimethylammonium, i.e., quaternary ammonium compounds with a 16-carbon alkyl chain.However, in a subsequent study, carried out by the same group of researchers (Lemke et al 2001) showed that clays modified with Hexadecyl trimethyl ammonium or with Hexadecyl amine not only did not show protection from the estrogenic effect of Zearalenone but even produced a greater estrogenic effect, since in this bioassay with female mice it was observed that the relative weight of the uterus with respect to the weight of the mice was greater in the group consuming the clay modified with Hexadecyl trimethyl ammonium than the group that consumed Zearalenone. For its part, document US20100330235 (2010) describes a mycotoxin adsorbent, for example, Aflatoxin, Zearalenone, Ochratoxin A and Fumonisin B1, based on the combination of an organic silicate with an amorphous structure and dodecylamine (a primary amine that has a linear, nonpolar, aliphatic, twelve-carbon carbon chain). Furthermore, we found in the prior art document W002052950 (2002), which describes a modified organomineral containing a long-chain quaternary amine, for example, dioctadecyldimethylamine, octadecyltrimethylamine, octadecyldimethylbenzylamine, and similar compounds. This modified organomineral is used as a feed additive to adsorb various mycotoxins in animals at a dose of 0.2%, but it does not include fumonisin B1, a mycotoxin very common in corn, which causes serious problems in livestock production, especially in pigs, cattle, and horses. Among the quaternary ammonium compounds most commonly used to organically modify aluminosilicates are benzalkonium chloride and didecyldimethylammonium chloride, which are widely used as biocides or disinfectants. However, they are currently subject to new regulations regarding their use, which have limited their residue levels in food. This stems from the fact that quaternary ammonium compounds, in addition to being used as biocides, have also been used as pesticides. The classification of quaternary ammonium compounds as pesticides has led to the establishment of a maximum residue limit (MRL) in food, which in 2014 was set at 0.1 mg / kg (Regulation (EU) 1119 / 2014). This situation calls into question the use of these quaternary ammonium compounds for the organic modification of aluminosilicates that are used as mycotoxin adsorbents in animal feed. One possible solution to the above is to use a quaternary ammonium compound that is not considered a pesticide. One of these possible compounds is dioctadecyldimethylammonium chloride. This quaternary ammonium compound is approved in the United States for use in the refining of sugar cane for human consumption (21CFR173.400) and in Europe for use as an organic modifier of montmorillonite used in plastics that come into contact with food (EFSA Journal 2015;13(11):4285). Dioctadecyldimethylammonium chloride has already been mentioned in document W002052950 (2002), discussed above; however, this document uses a clipnotilolite-type zeolite as the aluminosilicate, not a phyllosilicate such as bentonite. Furthermore, this document does not mention the efficacy of this material against fumonisin B1, the most prevalent mycotoxin in maize. Furthermore, in the document MRStockmeyer Adsorption of organic compounds on organophilic bentonites, Applied Clay Science, Volume 6, Issue 1, May 1991, pages 39-57, he discloses the use of dioctadecyldimethylammonium chloride to modify a bentonite but with the objective of adsorbing some organic compounds that pollute the environment such as phenol, aniline, nitroethane, diethyl ketone among others, but not mycotoxins, much less Fumonisin B1. As can be seen, in the state of the art Fumonisin continues to be a problem because, although it is reported to adsorb onto organically modified aluminosilicates, these are normally produced with didecyldimethylammonium chloride and benzalkonium chloride, which are subject to new regulations. Therefore, none of the mycotoxin adsorbents known in the prior art have used the substitution of dioctadecyldimethylammonium to organically modify a bentonite that is used and formulated in animal feed for the control of mycotoxins, especially Fumonisin B1 and Zearalenone, which is part of the object of the present invention. In this application, an organically modified bentonite with dioctadecyldimethylammonium chloride is used as an adsorbent of mycotoxins in animal feed. BRIEF DESCRIPTION OF THE INVENTION The inventors of this application have unexpectedly found that an organically modified phyllosilicate containing dioctadecyldimethylammonium chloride is useful as a mycotoxin adsorbent. Specifically, the use of an organically modified bentonite containing dioctadecyldimethylammonium offers high mycotoxin adsorption efficiency and can be safely used in animal feed since this quaternary ammonium compound is not considered a pesticide. In a first aspect of the present application, the invention relates to the use of a mycotoxin adsorbent employed in balanced feed to avoid the harmful effects of mycotoxins, particularly Fumonisin B1, since it was found that at a certain level of inclusion of the organic compound dioctadecyldimethylammonium in bentonite a higher percentage of Fumonisin B1 adsorption is obtained than at other inclusions. In another aspect, the invention relates to a premix for preparing the mycotoxin adsorbent, the use of an additive for animal feed, and an animal feed formulation that includes the mycotoxin adsorbent of the invention. The present invention provides a process for preparing a mycotoxin adsorbent by reacting an aluminosilicate base with a cation exchange capacity of between 30 milliequivalents / 100 g and 60 milliequivalents per 100 g of material with dioctadecyldimethylammonium chloride. Furthermore, the present invention relates to the use of the organically modified bentonite containing dioctadecyldimethylammonium chloride of the invention for addition to feed contaminated with mycotoxins. The invention also relates to a method for preparing balanced animal feed that prevents mycotoxicosis problems in animals. The mycotoxin adsorbent is used by adding it to the contaminated food, either in granular or powder form, for better adsorption of the mycotoxins, which are then eliminated together through the feces. Thus, by applying the present invention, the absorption of mycotoxins from contaminated feed into the animal's gastrointestinal tract is prevented. This substantially improves animal health, resulting in better zootechnical performance. DETAILED DESCRIPTION The present invention is based on the fact that a modification of the surface of phyllosilicates, particularly bentonite, by means of a long-chain quaternary ammonium compound can be used for the adsorption of mycotoxins and especially for the adsorption of Fumonisin and Zearalenone. In particular, in the present invention the modification of the surface of a phyllosilicate such as bentonite is carried out with dioctadecyldimethylammonium chloride, a compound that, not being used as a pesticide, does not have the same restrictions as other long-chain quaternary ammonium compounds such as didecyldimethylammonium chloride or benzalkonium chloride regarding a residue limit in food. Dioctadecyldimethylammonium chloride, which is used for phyllosilicate surface modification, can occupy some or all of the active sites on the aluminosilicate surface. The aluminosilicate used is a phyllosilicate such as bentonite, provided that the material used has a cation exchange capacity of at least 30 milliequivalents per 100 grams of material, in the range between 30 milliequivalents per 100 grams to 60 milliequivalents per 100 grams of material and preferably 60 milliequivalents per 100 grams of material. Therefore, one embodiment of the present invention consists of an organically modified phyllosilicate with dioctadecyldimethylammonium chloride for use as a mycotoxin adsorbent, wherein the phyllosilicate has a cation exchange capacity between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g. Preferably, said phyllosilicate is a bentonite, and more preferably the bentonite has a cation exchange capacity of 60 milliequivalents per 100 grams of material. In another aspect of this modality, the invention consists of the use of an organically modified phyllosilicate with dioctadecyldimethylammonium chloride as a mycotoxin adsorbent, wherein the phyllosilicate has a cation exchange capacity between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g. Preferably, said phyllosilicate is a bentonite and more preferably the bentonite has a cation exchange capacity of 60 milliequivalents per 100 grams of material. Another aspect of this modality includes the use of an organically modified phyllosilicate with dioctadecyldimethylammonium chloride to manufacture a balanced animal feed formulation for adsorbing mycotoxins, wherein the phyllosilicate has a cation exchange capacity between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g. Preferably, said phyllosilicate is a bentonite and more preferably the bentonite has a cation exchange capacity of 60 milliequivalents per 100 grams of material. ΐνίΛ / α / ^υ^ο / υΊ Ó4Z0 Dioctadecyldimethylammonium chloride is used at a ratio of 60% to 115% of the cation exchange capacity of the phyllosilicate used, in this case bentonite. The reaction is carried out in an aqueous medium with stirring at an initial temperature of 65°C to allow dissolution of the quaternary ammonium compound, with a reaction time of 1 hour. The product is separated by filtration, dried at a temperature between 85 and 140°C, and granulated or ground to a mesh size between 100 and 200. Preferably, dioctadecyldimethylammonium chloride is used at a ratio of 71% of the cation exchange capacity, when the phyllosilicate is bentonite. The use of the additive mentioned in the present invention is a low inclusion mycotoxin adsorbent that is added to mycotoxin-contaminated food at a rate of 0.05% to 0.15% of the food weight. Therefore, one embodiment of the present invention corresponds to a balanced animal feed formulation that includes a mycotoxin adsorbent of the present invention, characterized in that the adsorbent is used in doses of 0.05% to 0.15% by weight of the feed. In another aspect of this modality, the invention consists of a balanced animal feed formulation that includes an organically modified phyllosilicate with dioctadecyldimethylammonium chloride of the present invention, characterized in that the adsorbent is used in doses of 0.05% to 0.15% by weight of the feed. A preferred embodiment of the present invention relates to the use of a mycotoxin adsorbent characterized in that it comprises an organically modified bentonite with dioctadecyldimethylammonium chloride, wherein the phyllosilicate has a cation exchange capacity between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g, and wherein the dioctadecyldimethylammonium chloride is used in a proportion of 60% to 115% of the cation exchange capacity of the bentonite. In a preferred embodiment, the invention consists of an organically modified phyllosilicate with dioctadecyldimethylammonium chloride for use as a mycotoxin adsorbent, wherein the phyllosilicate has a cation exchange capacity between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g, wherein the dioctadecyldimethylammonium chloride is used in a proportion of 71% of the cation exchange capacity of the phyllosilicate. The invention also relates to a premixture for preparing an additive for balanced animal feed comprising the organically modified phyllosilicate with dioctadecyldimethylammonium chloride in accordance with the present invention. The invention further consists of the use of an animal feed additive comprising a mycotoxin adsorbent, a premix for preparing a mycotoxin adsorbent, or an animal feed additive to treat or prevent one or more adverse effects or reproductive symptoms associated with mycotoxin poisoning such as Fumonisin B1 and Zearalenone. Therefore, an additional modality relates to a balanced animal feed formulation that includes an organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to the present invention, for use in treating or preventing one or more harmful effects or symptoms in the digestive tract associated with mycotoxin poisoning, selected from the group that includes Aflatoxin B1, Ochratoxin A, Fumonisin B1, and Zearalenone. One aspect of this modality relates to the use of an organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to the present invention, to manufacture a balanced animal feed formulation to treat or prevent one or more adverse effects or symptoms in the digestive tract associated with mycotoxin poisoning, selected from the group that includes Aflatoxin Bl, Ochratoxin A, Fumonisin B1 and Zearalenone. IVIA / a / ZUZÓ / UI Ó4Z0 This application also relates to a process for preparing the organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to the present invention, said process being characterized by the following steps: a) Heat water to 65°C and add dioctadecyldimethylammonium chloride to dissolve it and form a solution, wherein the amount of dioctadecyldimethylammonium chloride must be equivalent to react with 60% or 115% of the cation exchange capacity of the phyllosilicate used; b) In another container, put water at room temperature and disperse in it the phyllosilicate with a cation exchange capacity of between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g of material in an aqueous medium with agitation; c) Transfer the phyllosilicate suspension formed in step (b) to the dioctadecyldimethylammonium chloride solution formed in step (a) and stir while maintaining heating at a temperature of approximately 65°C for 15 minutes, then stop heating but continue stirring for another 45 minutes. The suspension is expected to reach room temperature at the end. d) separate by filtration and dry at a temperature between 40 and 150°C; and e) grind to a 200 mesh particle size or granulate it. In particular, the phyllosilicate in the process for preparing the organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to the present invention is bentonite. Preferably, in the process for preparing the organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to the present invention, the dioctadecyldimethylammonium chloride is used in a proportion of 60% to 115% of the cation exchange capacity of the phyllosilicate used and, more preferably, the dioctadecyldimethylammonium chloride is used in a proportion of 71.0% of the cation exchange capacity and the phyllosilicate is bentonite. The following examples are provided for the purpose of describing the best known method or the best way envisaged by the applicant to implement the invention, as well as information supporting the industrial application of the present invention, and not to limit the scope of the claims. Example 1 FIRST PREPARATION OF THE MYCOTOXIN ADSORBENT AND ITS IN VITRO EVALUATION. 114.8% replacement of the cation exchange capacity. For this synthesis and in vitro evaluation to determine its feasibility as a mycotoxin adsorbent, a commercial dioctadecyldimethylammonium chloride (DODAC) product with a molecular weight of 586 and an active ingredient concentration of 75% by weight was used. The bentonite used has a cation exchange capacity of 60 milliequivalents per 100 g. Since DODAC has low solubility in water, the process requires heating the water. The specific synthesis procedure was: a) Dissolve 35 g of DODAC in 350 mL of hot water at 65 °C in a beaker. Stir for 15 minutes. Use a heating plate with stirring for heating. b) In another beaker, 65 g of bentonite are placed in 150 mL of water. It is stirred for 15 minutes to make a suspension. c) Transfer the bentonite to the beaker with the DODAC and stir, maintaining the temperature at 65 °C for 15 more minutes. Then turn off the heat and continue stirring for 45 minutes. Finally, wait until it reaches room temperature. d) Filter the suspension and let it rest for two hours. e) Recover all solid material and dry at 135 °C until the material is dry. f) Grind to 200 mesh. With the mass proportions used in this first synthesis, there is a substitution by the quaternary of 114.8% of the cation exchange capacity of bentonite of 60 milliequivalents per 100 g. The material thus obtained was subjected to an in vitro evaluation of its individual adsorption capacity for four of the main known mycotoxins: Aflatoxin B1, Zearalenone, Ochratoxin A, and Fumonisin B1. The concentrations of the mycotoxins used were equivalent to 3000 ppb of each, with an adsorbent dosage of 0.15%. Adsorption was carried out at pH 3 and desorption at pH 6.5. The difference between adsorption and desorption gives the adsorption efficiency. The results obtained for this sample are presented below: % Adsorption % Desorption % Efficiency Aflatoxin B1 95 4 91 Zearalenone 94 1 93 Ochratoxin A 97 1 96 Fumonisin B1 78 6 These results show very high efficiency values, except for Fumonisin B1. Example 2 SECOND PREPARATION OF THE MYCOTOXIN ADSORBENT AND ITS IN VITRO EVALUATION. 102.6% replacement of the cation exchange capacity. A second synthesis and evaluation of the mycotoxin adsorbent was carried out. This synthesis was performed to try to obtain better efficiency for Fumonisin B1. For this second synthesis, the same procedure as in example 1 was followed, where the amount of DODAC was decreased to 32.5 g and the amount of bentonite was adjusted to 67.5 g. With these proportions, a substitution of 102.6% of the cation exchange capacity of bentonite is obtained. The in vitro evaluation of this mycotoxin adsorbent is presented below. % Adsorption % Desorption % Efficiency Aflatoxin B1 96 3 93 Zearalenone 97 1 96 Ochratoxin A 98 0 98 Fumonisin B1 80 0 80 This modification improved the adsorption efficiency of Fumonisin Bl. Example 3 THIRD PREPARATION OF THE MYCOTOXIN ADSORBENT AND ITS IN VITRO EVALUATION. 91.3% replacement of the cation exchange capacity. A third synthesis and evaluation of the mycotoxin adsorbent was carried out. This synthesis was performed to try to observe, contrary to all expectations, whether a decrease in the active ingredient and consequently a decrease in the substitution of the cation exchange capacity would improve the adsorption efficiency for Fumonisin B1 without affecting the adsorption of the other mycotoxins, mainly Zearalenone. For this third synthesis, the same procedure as in example 1 was followed, where the amount of DODAC was decreased to 30.0 g and the amount of bentonite was increased to 70.0 g. With these proportions, a 91.3% substitution of the cation exchange capacity of bentonite is achieved. The in vitro evaluation of this mycotoxin adsorbent is presented below. % Adsorption % Desorption % Efficiency Aflatoxin B1 96 4 92 Zearalenone 99 1 98 Ochratoxin A 99 0 99 Fumonisin B1 90 1 89 Improved efficiency was obtained for Fumonisin B1 without affecting the efficiency of the other mycotoxins. Example 4 FOURTH PREPARATION OF THE MYCOTOXIN ADSORBENT AND ITS IN VITRO EVALUATION. 80.8% replacement of the cation exchange capacity. In the fourth experiment, the concentration of the active ingredient DODAC was reduced again, which would lead to a further reduction in cost. In this experiment, the amount of DODAC was decreased to 27.5 g and the amount of bentonite was increased to 72.5 g. With these proportions, an 80.8% substitution of the bentonite's cation exchange capacity was achieved. In the in vitro evaluation, shown in the table below, it can be seen that the adsorption efficiency of Zearalenone is maintained and the adsorption of Fumonisin B1 is increased. % Adsorption % Desorption % Efficiency Phiatoxin B1 93 5 88 Zearalenone 100 0 100 Ochratoxine A 100 0 100 Fumonisin B1 95 i 94 Example 5 FIFTH PREPARATION OF THE MYCOTOXIN ADSORBENT AND ITS IN VITRO EVALUATION. 71.0% replacement of the cation exchange capacity. A fifth experiment was conducted, following the unexpected observation that decreasing the percentage of substitution increased the adsorption efficiency of Fumonisin B1, without affecting the adsorption efficiency of the other mycotoxins. To carry out the synthesis, the active ingredient DODAC was reduced to 25 g and the bentonite was increased to 75 g. With these proportions, a 71.0% substitution of the cation exchange capacity of bentonite is obtained, with a cation exchange capacity of 60 milliequivalents per 100 g. The results were surprisingly superior as Fumonisin B1 is shown below, especially in the case of % Adsorption % Desorption % Efficiency Aflatoxin B1 97 3 94 Zearalenone 100 1 99 Ochratoxin A 97 1 96 Fumonisin B1 100 3 97 Example 6. IN VIVO EVALUATION OF THE EFFICACY OF THE MYCOTOXIN ADSORBENT IN CONTROLLING THE ESTROGENIC EFFECTS OF ZEARALENONE. With the aim of verifying the effectiveness of the prototype of example 5, identified as DODAC, since it was the one that presented the best in vitro results, an in vivo evaluation was carried out using pre-pubertal female pigs to study the effectiveness against the estrogenic effect of Zearalenone and with contamination of Deoxynivalenol and Fumonisin B1. Thirty-two recently weaned female pigs, 21 days old, were used and distributed into three treatments with two female pigs per replicate. The negative control group consumed feed with low mycotoxin content and without the addition of the DODAC prototype. The positive control group included feed contaminated with high levels of mycotoxins, primarily zearalenone, and did not include the prototype. The DODAC treatment included feed contaminated with high levels of mycotoxins and 0.15% by weight of the DODAC prototype feed. The negative and positive control treatments had five replicates each, and the challenge treatment had six replicates. The first seven days were for adaptation. The experimental diets were as follows: Treatment DODAC (g / kg) Zearalenone (pg / kg) Deoxynivalenol (pg / kg) Fumonisin B1 (pg / kg) Negative control 0 81 < 20 994 Positive control 0 395 660 1115 DODAC Challenge 1.5 391 555 1065 Zearalenone was obtained from a natural contamination with the fungus Fusarium graminearum. The feed was commercially available. The concentration of ZEA added to this feed was verified in the laboratory by HPLC. Female pigs were received, weighed, and placed in their pens for an adaptation period (7 days). Subsequently, they were weighed again and distributed in groups of two animals per replicate, with their respective treatment. Individual weight was recorded weekly until the end of the experiment. Feed conversion ratio, feed intake, weight gain, and vulva measurements (length x width x depth) were calculated weekly. > tü r\ c K.1a -t; N a Because the estrogenic effect of ZEA manifests as inflammation, reddening of the vulva, and growth of the reproductive tract, these parameters were considered to measure ZEA toxicity. At the end of the experiment, the females were sacrificed; subsequently, their reproductive tract, liver, kidneys, heart, lung, and spleen were removed, weighed individually to obtain relative weight, and samples were taken for histopathological testing. The results obtained after 21 days of experimentation showed no statistically significant differences between treatments in weight gain and feed consumption, only in feed conversion (Table 1). Table No. 1 Initial Weight, Final Weight (21 days), Weight Gain, Feed Conversion and Feed Intake. Treatments Weight < 3 in g Weight gain Feed conversion Feed intake Means ± standard error Means ± standard error Means ± standard error Means ± standard error Means ± standard error Means ± standard error Negative control 5640 ±190 a 14430 ±547 a 8790 ± 425 a 1.46 ±0.030 a 12800 ± 566 a Positive control ZEA 5790 ±164 a 14060 ± 419 a 8270 ± 309 a 1.60 ± 0.030 b 13260 ± 449 a DODAC + ZEA 6200 ± 179 a 15010 ± 316 a 8810 ± 286 a 1.54 ± 0.037 ab 13570 ±298 a Means with different letters are statistically significant for p < 0.05. Regarding reproductive parameters, the negative effect of Zearalenone was observed, with statistically significant differences from the first week until the end of the experiment. The inclusion of prototype 5 DODAC improved the evaluated parameters compared to the positive control (Table 2). Table No. 2 Vulva volume (cm3 / kg), percentage (%) of the relative weight of the vulva and reproductive system at the end of the test. PR Treatments Reproductive System (%) PR Vulva (%) Vulvar Volume (cm3 / kg) Means ± standard error Means ± standard error Means ± standard error Negative Control 0.0822 ± 0.0041 a 0.0226 ♦ 0.0019 a 0.100 ± 0.013 a Positive Control ZEA 0.1597 ± 0.0077 c 0.0483 ± 0.0024 c 0.364 ± 0.027 c DODAC + ZEA 0.1182 ± 0.0055 b 0.0313 ± 0.0014 b 0.189 ± 0.014 b DODAC Effectiveness 53.5% 66.1% 66.3% Means with different letters are statistically significant for p < 0.05. The sum total of the effectiveness on the reproductive system resulted in a 61.9% benefit when using the prototype from Example 5, identified as DODAC 15, for 21 days on a diet contaminated with 300 ppb of zearalenone, higher than the concentration in the negative control diet. The relative weights of the organs (liver, kidney, spleen) were not affected by zearalenone consumption.

Claims

1. An organically modified phyllosilicate with dioctadecyldimethylammonium chloride for use as a mycotoxin adsorbent, wherein the phyllosilicate has a cation exchange capacity between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g.

2. The phyllosilicate organically modified with dioctadecyldimethylammonium chloride for use according to claim 1, wherein the phyllosilicate is a bentonite.

3. The organically modified phyllosilicate with dioctadecyldimethylammonium chloride for use according to claim 2, wherein the bentonite has a cation exchange capacity of 60 milliequivalents per 100 g.

4. The phyllosilicate organically modified with dioctadecyldimethylammonium chloride for use in accordance with any of claims 1 to 3, wherein the dioctadecyldimethylammonium chloride is used in a proportion of 60% to 115% of the cation exchange capacity of the phyllosilicate used.

5. The phyllosilicate organically modified with dioctadecyldimethylammonium chloride for use according to any of claims 1 to 4, wherein the dioctadecyldimethylammonium chloride is used in a proportion of 71.0% of the cation exchange capacity of the bentonite.

6. A premixture for preparing an animal feed additive comprising 26-dioctadecyldimethylammonium chloride organically modified phyllosilicate according to any of claims 1 to 5.

7. A balanced animal feed formulation that includes the organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to any of claims 1 to 5, characterized in that the organically modified phyllosilicate with dioctadecyldimethylammonium chloride is used in doses of 0.05% to 0.15% by weight of the feed.

8. A formulation according to claim 7, for use in treating or preventing one or more adverse effects or symptoms in the digestive tract associated with mycotoxin poisoning, selected from the group including Aflatoxin B1, Ochratoxin A, Fumonisin B1, and Zearalenone.

9. A process for preparing the organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to any of claims 1 to 5, said process characterized by the following steps: a) heating water to 65°C and adding dioctadecyldimethylammonium chloride to dissolve it and form a solution, wherein the amount of dioctadecyldimethylammonium chloride is equivalent to react with 60% or 115% of the cation exchange capacity of the phyllosilicate used; b) in another container, placing water at room temperature and dispersing the phyllosilicate with a cation exchange capacity of between 30 milliequivalents per 100 g and 60 milliequivalents per 100 g of material in an aqueous medium with stirring;c) Transfer the phyllosilicate suspension formed in step (b) to the dioctadecyldimethylammonium chloride solution formed in step (a) and stir, maintaining heat at approximately 65°C for 15 minutes, then stop heating but continue stirring for another 45 minutes. The suspension should eventually reach room temperature. d) Separate by filtration and dry at a temperature between 40 and 150°C; and e) Grind to a 200 mesh size or granulate.

10. The process for preparing the organically modified phyllosilicate with dioctadecyldimethylammonium chloride according to claim 9, wherein the phyllosilicate is bentonite and wherein the dioctadecyldimethylammonium chloride is used in a proportion of 71.0% of the cation exchange capacity.