Animal feed product comprising dehydrated, activated and ground diatomaceous earth with p-cymenol, allicin and carvacrol, method for producing same and use thereof

Abstract

The present invention relates to an animal feed product comprising dehydrated, activated and ground diatomaceous earth in combination with p-cymenol, allicin and carvacrol, as well as to its method of production and use thereof as animal feed. The product, which combines only naturally sourced products, leads to enhanced productivity on animal breeding farms when added to the feed of said animals, by combining the ability to reduce the rate of forward movement of the feed in the digestive tract provided by the diatomaceous earth with the ability to physiologically stimulate enterocytes and the enhanced control of pathogen microorganisms provided by p-cymenol, allicin and carvacrol.

Description

[0001] DESCRIPTION

[0002] ANIMAL FEED PRODUCT COMPRISING DEHYDRATED, ACTIVATED AND GROUND DIATOMACEOUS EARTH WITH P-CIMENOL, ALLICIN AND CARVACROL, PROCEDURE FOR ITS OBTAINING AND USE

[0003] Technical field

[0004] The present invention relates to the field of animal feed. In particular, it relates to an animal feed product comprising dehydrated, activated, and ground diatomaceous earth in combination with p-cimenol, allicin, and carvacrol, and the process for obtaining it.

[0005] Background of the invention

[0006] A well-regulated intestinal transit speed is essential for feed digestion and nutrient absorption [2]-[4], processes that allow high production performance in poultry and pig farms [3], [5]. In this sense, rapid transit disorders should be avoided because they cause the feed to remain in the intestinal lumen for less time and not to have sufficient contact with the digestive enzymes [4] and intestinal villi responsible for nutrient assimilation [2]. Ultimately, the feed is not assimilated correctly and, despite the consumption of large quantities of feed, the use of the diet is worsened and feed costs on farms increase.

[0007] It is also known that, in animal feed, high amounts of undigested proteins reach the intestine; these proteins cannot be absorbed and, on the other hand, are used by pathogenic bacteria that metabolize them. This causes bacterial populations to increase, reducing the population of intestinal flora. Consequently, the intestine will not function properly, and in some cases, diarrhea may even occur. It would be useful to have an animal feed product that would increase the time food remains in the digestive tract and, in addition, help achieve better control of pathogenic microorganisms in the intestine.

[0008] The present invention provides a solution to that problem.

[0009] Summary description of the invention

[0010] A first object of the present invention relates to an animal feed product comprising dehydrated, activated, and ground diatomaceous earth in combination with p-cimenol, allicin, and carvacrol, as defined in the claims. Since this product comprises several components, it is also referred to throughout this specification as the composition of the invention.

[0011] A second object of the present invention relates to a process for obtaining a product as defined in the first object of the present invention and in the claims.

[0012] A third object of the present invention is the use of the product as defined in the first object of the present invention for feeding animals, preferably poultry and pigs.

[0013] Brief description of the figures

[0014] Fig. 1. Molecular structure of p-cimenol.

[0015] Fig. 2. Molecular structure of allicin.

[0016] Fig- 3. Molecular structure of carvacrol.

[0017] Fig- 4. Structure of a diatom of the genus Thalassiosira.

[0018] Fig- 5. Structure of a diatom of the genus Actinoptychus.

[0019] Detailed description of the invention The present invention provides a novel product that uses diatomaceous earth as a carrier of the three active ingredients p-cimenol, allicin and carvacrol, which stimulate the activity of enterocytes, cells of the digestive mucosa.

[0020] Diatomaceous earth is made from diatoms, the exoskeletons of unicellular algae that have settled over millions of years. Their cell walls are made of opaline silica (hydrated silicon dioxide). The structure of diatoms, composed of two valves with pores on their surface and a space between them, has a high absorption capacity. Their use as transporters or carriers has been described [1]. Due to their structure, diatoms are capable of absorbing certain components and releasing them over a more or less prolonged period within the digestive tract of animals. As the active ingredients are released, the diatoms are freed and are able to reduce the rate at which food moves through the digestive tract.

[0021] p-Cimenol, allicin and carvacrol, on the other hand, physiologically stimulate the activity of enterocytes, cells that make up the intestinal mucosa, thus optimizing their functions related to digestive processes and the absorption of nutrients [6]-[9]. They also help to achieve greater control of pathogenic microorganisms in the intestine, which decreases the incidence of clinical and subclinical diseases, thanks to the improvement of the state and functions of enterocytes [7]—

[0010] .

[0022] Thus, the combination of p-cimenol, allicin, and carvacrol with diatomaceous earth allows for the creation of an invention with a significant capacity to improve the condition of the intestinal mucosa and an effect that lasts for a long time, thanks to the diatoms gradually releasing the three active ingredients within the digestive tract.

[0023] Ultimately, the invention improves the productivity of animal breeding farms [8],

[0011] ; this is observed by an improved feed conversion ratio (ratio of weight gained to feed consumed) and growth rate (grams of daily weight gain), thanks to improved intestinal well-being [7],

[0010] . Furthermore, the food product is prepared by combining only products of natural origin: diatomaceous earth and plant extracts (Thumus vulgaris, Allium sativum and Origanum vulgare) concentrated, respectively, in p-cimenol, allicin and carvacrol.

[0024] Thus, in a first aspect, the present invention relates to an animal feed product comprising dehydrated, activated, and ground diatomaceous earth in combination with p-cimenol, allicin, and carvacrol. Preferably, the product is in powder form.

[0025] Furthermore, the present inventors have developed a process for obtaining the product of the invention that allows obtaining a powdered product that is easy to store and administer, a process that constitutes a second aspect of the invention.

[0026] Furthermore, the use of the food product of the first aspect of the invention as an additive in animal feed is also part of the invention and constitutes a third aspect thereof. Preferably, the animals are selected from the group of mammals and birds used in animal production. Thus, the use of the invention includes, but is not limited to, pigs among the mammals, and broiler chickens, laying hens, breeding hens, turkeys, and ducks among the birds used in animal production.

[0027] Dehydrated, activated and ground diatomaceous earth

[0028] In a preferred embodiment, the diatomaceous earth in the food product, which comprises dehydrated, activated, and ground diatomaceous earth, has a minimum content of 60% diatoms of the genera Thalassiosira and Actinoptychus relative to the total diatoms in the additive. In another possible embodiment, compatible with the above, the dehydrated, activated, and ground diatomaceous earth is present in at least 50% by weight relative to the total weight of the food product composition.

[0029] The dehydrated, activated and ground diatomaceous earth acts, firstly, as a transporter or carrier for the rest of the components of the invention and, secondly, as the components of the invention are released into the intestinal lumen, as a slower of intestinal transit.

[0030] This diatomaceous earth undergoes selection, dehydration, activation, and grinding processes to obtain diatomaceous earth with a high volume-to-surface ratio and eliminate water content, thereby increasing its capacity to absorb p-cimenol, allicin, and carvacrol.

[0031] As used in this application, dehydrated diatomaceous earth is defined as earth in which at least 80% by weight of the water content, preferably at least 95%, has been removed. This can be achieved, for example, by heating to temperatures above 100°C, as explained below.

[0032] On the other hand, activated diatomaceous earth is understood to be that in which both the pores and the silanol groups on the surface of the diatoms have been freed of water and other possible impurities, which can be achieved, as explained below, by heating to a temperature above 100°C, so that the heating that serves for dehydration is also useful for activation.

[0033] As for ground diatomaceous earth, it is understood as having an average particle size of less than 0.2 - 5 mm, preferably less than 125 pm. More preferably, for the purposes of the invention, in ground diatomaceous earth at least 95% of the particles have a size between 50 pm and 125 pm. p-cimenol

[0034] In another preferred embodiment, p-cimenol (Fig. 1) is present in at least 2.1% by weight of the composition of the present invention. For the purposes of the present invention, said compound is preferably contained in a concentrated extract of Thymus vulgaris, which forms part of the composition of the present invention. More preferably, the Thymus vulgaris extract comprises p-cimenol in a weight percentage between 14% and 16% with respect to the total weight of said extract and the extract is present in the composition in a percentage of 15% by weight with respect to the total weight of the composition of the invention. p-cimenol is a monoterpene whose functions as a bactericide, fungicide and preservative are known

[0012] ,

[0013] and is present in many botanical species, including Thymus vulgaris.

[0035] In vitro assays and field trials indicate that p-cimenol is effective against pathogenic microorganisms such as Escherichia coli, Salmonella, Staphylococcus aureus

[0013] , Aspergillus, Penicillium, Fusarium and Candida

[0013] ,

[0014] ,

[0036] Contact with p-cimenol causes, in pathogenic bacteria and fungi, the immediate release of cellular contents into the medium, caused by the perforation of the bacterial membrane that leads to the destruction of the cell by osmotic shock

[0012] , This is due to the interaction of p-cimenol with the lipids that make up the cytoplasmic membrane

[0013] , In fungi, p-cimenol also acts by inhibiting the synthesis of ergosterol

[0014] , the most important sterol in its cell membrane, and causing morphological alterations in the hyphae

[0015] ,

[0037] Allicin

[0038] In another preferred embodiment, allicin (Fig. 2) is present in at least 3.25% by weight with respect to the total weight of the composition of the present invention. For the purposes of the present invention, said compound is preferably contained in a concentrated extract of Allium sativum that forms part of the composition of the present invention. More preferably, the Allium sativum extract contains allicin in a percentage by weight between 13% and 14% with respect to the total weight of said extract, and the extract is present in the composition in a percentage of 25% by weight with respect to the total weight of the composition of the invention.

[0039] Allicin is a thiosulfate produced from the reaction of alliin with the enzyme alliinase, after grinding garlic (the bulb of Allium sativum, which is the part generally used for food) and breaking down its tissues

[0016] ,

[0017] , It promotes antioxidant and anti-inflammatory activity and has a positive effect on digestive functions, promoting peristalsis and digestive emptying, helps regenerate damaged digestive tissue and promotes the activity of digestive enzymes

[0018] , It also has antimicrobial action against various pathogenic bacteria in animals such as E. coli, Staphylococcus spp., Salmonella spp. and Enterococcus

[0016] ,

[0017] , and fungi such as Candida, Aspergillus and Penicillium

[0016] , since it acts on the integrity of the fungal cell wall and the organelles of microorganisms.

[0040] Carvacrol

[0041] In another preferred embodiment, carvacrol (Fig. 3) is present in at least 1.7% by weight relative to the total weight of the composition of the present invention. For the purposes of the present invention, said compound is preferably contained in a concentrated extract of Origanum vulgare that forms part of the composition of the present invention. More preferably, the extract of Origanum vulgare contains carvacrol in a percentage by weight between 17% and 20% relative to the total weight of said extract, and the extract is present in the composition in a percentage of 10% by weight relative to the total weight of the composition of the invention.

[0042] Carvacrol is a phenolic monoterpene that has antimicrobial action

[0013] ,

[0015] ,

[0019] by increasing the permeability of the cytoplasmic membrane of bacteria and inhibiting the ATPase enzyme, necessary to perform biological functions

[0020] , It also has antioxidant activity, which favors the functioning of the digestive system of animals

[0019] ,

[0043] Method of the present invention

[0044] In a second aspect, the present invention relates to a process for obtaining an animal additive, according to the first aspect of the invention, that is, an animal additive comprising dehydrated, activated and ground diatomaceous earth in combination with p-cimenol, allicin and carvacrol, comprising the following steps:

[0045] (a) obtaining a diatomaceous earth with a minimum content of 60% diatoms of the genera Thalassiosira (fig. 4) and Actinoptychus (fig. 5) with respect to the total quantity of diatoms;

[0046] (b) activating and dehydrating the diatomaceous earth; (c) grinding the diatomaceous earth obtained in step (b);

[0047] (d) obtaining a concentrated extract of p-cimenol from the leaves of Thymus vulgaris,'

[0048] (e) obtaining an extract concentrated in allicin from bulbs of Allium sativum,'

[0049] (f) obtaining a concentrated extract of carvacrol from the leaves of Origanum vulgare c,

[0050] (g) mixing the diatomaceous earth from step (c) with the extracts from steps (d), (e), and

[0051] (f) until homogeneity.

[0052] Preferably, step (a) comprises the following sub-steps:

[0053] (al) taking at least one series of at least 10 samples of a diatomaceous earth to be evaluated;

[0054] (a2) determine under optical microscope in each of the samples, the percentage of diatoms of the genera Thalassiosira and Actinoptychus;

[0055] (a3) select a diatomaceous earth where 100% of the observed samples comprise at least 60% diatoms of the genera Thalassiosira and / or Actinoptychus.

[0056] Preferably, in sub-step (al) 10 samples of diatomaceous earth are examined in each sample series. Preferably, at least two series of at least 10 samples each are examined. Also preferably, each of the samples weighs in the range of 0.9-1.1 grams, with 1 gram samples being a suitable embodiment.

[0057] Sub-step a2) of determining the composition of diatomaceous earth is carried out based on the symmetry of the frustules or silica shells, which must be radial in both genera, with the pores distributed evenly over the entire surface, as in Fig. 4 (genus Thalassiosira) or radial, as in Fig. 5 (genus Actinoptychus). The total percentage of diatoms belonging to one genus or another is calculated by identifying and counting the number of diatoms of each genus in each sample and calculating their percentage of the total number of diatoms in the sample. In the selected diatomaceous earth, with a minimum of 60% diatoms of the genera Thalassiosira and / or Actinoptychus, the remaining percentage (maximum 40%) is made up of other diatoms.Preferably, step (b) is carried out by exposure to temperatures between 150°C and 225°C, preferably between 190°C and 210°C, more preferably at 200°C, for between 25 and 30 minutes.

[0058] This temperature range is sufficient to activate the diatoms, keeping them intact, and evaporate more than 95% of the water content.

[0059] Optionally, it can be confirmed under a light microscope that the structure of the diatoms remains intact after heat treatment.

[0060] Activating diatomaceous earth removes water molecules and most of the impurities present on the surface and pores of the diatoms it contains. The goal of this activation is, therefore, to release the pores and silanol groups from the surface of the diatoms to increase their water-binding capacity.

[0061] Preferably, step (c) comprises the following sub-steps:

[0062] (el) grinding the activated dehydrated diatomaceous earth in a crushing and compression process;

[0063] (c2) Sieve the ground diatomaceous earth so that at least 95% passes through a sieve size of 125 pm opening.

[0064] The crushing process can be carried out using grooved rollers and reduces the average particle size to between 0.2 and 5 mm. The compression process can be carried out using smooth rollers and produces average particle sizes between 50 and 150 µm, with at least 95% being less than 125 µm. The size values ​​given here refer to the particle dimension with the greatest width.

[0065] This results in dehydrated and activated diatomaceous earth with at least 60% diatoms of the genera Thalassiosira and Actinoptychus and a particle size of less than 125 pm.

[0066] This dehydrated, activated and ground diatomaceous earth is present in at least 50% by weight of the composition of the present invention and will be used as a transporter or carrier for the rest of the components of the product of the invention and as a slower of intestinal transit, as the components of the invention are released.

[0067] Preferably, step (d) comprises the following sub-steps:

[0068] (di) cold press the leaves of Thymus vulgaris,'

[0069] (d2) suspend the extract obtained in (di) in water;

[0070] (d3) distill the oil from the leaf extract;

[0071] (d4) extracting the oil from the aqueous phase;

[0072] (d5) extract the phenolic content of the oil with a base and solubilize it in water;

[0073] (d6) extract p-cimenol with ether;

[0074] (d7) evaporate the ether.

[0075] In this way, a concentrated extract of p-cimenol is obtained. Example 1 of this application shows a more specific embodiment of said step d) comprising the steps (di) to (d7) above.

[0076] Preferably, step (e) comprises the following sub-steps:

[0077] (el) allowing allicin to be formed from alliin by the enzyme alliinase by macerating garlic powder with an aqueous solution of ethyl p-hydroxybenzoate and allowing to stand at room temperature;

[0078] (e2) centrifuge at 3500 rpm for 10 minutes;

[0079] (e3) recover the supernatant and add citric acid to stop the enzymatic catalysis reaction;

[0080] (e4) filter the solution obtained in (e3), recovering the filtrate obtained.

[0081] In this way, the allicin extract is obtained. Example 2 shows a particular embodiment of said step (e) with substeps (e1) to (e4).

[0082] Preferably, step (f) comprises the following sub-steps:

[0083] (fl) grinding and drying the Origanum vulgare leaves to obtain a powdered product;

[0084] (f2) macerate the powder obtained in (fl) in ethanol, preferably for 4 hours;

[0085] (f3) solidify the oleoresin obtained in (f2) by absorption with silica gel and evaporate the solvent, preferably maintaining the same temperature; (f4) centrifuge at 6000 rpm for 30 minutes;

[0086] (f5) Discard the aqueous phase.

[0087] This is how the carvacrol extract is obtained.

[0088] Use of the product of the invention for animal feed

[0089] As previously discussed, one aspect of the invention is the use of the product of the invention for animal feed. Preferably, the animals are either mammals, such as pigs, or birds. The birds are preferably birds commonly used in animal production and may be, for example, but not limited to, broiler chickens, laying hens, breeder hens, turkeys, and ducks.

[0090] Preferably, the feed product of the invention is added to the feed supplied to the animals, by mixing it with said feed. In the case of pigs, the composition of the invention can be added to their feed, as one possible embodiment, in an amount of 0.5 kg per ton of feed.

[0091] Examples

[0092] Example 1, Obtaining p-cimenol from an extract of Thymus vulgaris

[0093] To obtain the concentrated extract of p-cimenol, the leaves intended for thyme extract (Thymus vulgaris) were first cold pressed and ground until they became a homogeneous powder.

[0094] To obtain a higher concentration of active ingredient, the crushed leaves of Thymus vulgaris were placed in a flask with enough water to achieve the suspension of the sample, with a proportion of 1 kg of dry T vulgaris in 3 liters of water.

[0095] Minimal sample manipulation was sought to avoid degrading the oils, which are the main source of the compound to be purified. Therefore, steam distillation was used to extract the T. vulgaris oil. Because it is a highly volatile oil and insoluble in water, the product was allowed to form two phases. Subsequently, water extraction was performed to separate the pure oil using a separatory funnel. The oil was separated into a covered container and protected from light to prevent light decomposition.

[0096] The aqueous portion was further extracted with ether to separate the excess oil that remained solubilized in this phase. Traces of water were extracted from the ether phase using sodium sulfate as a drying agent. An acid-base extraction was performed to reduce the amount of components to be separated by chromatography.

[0097] To obtain p-cimenol, one of the most acidic compounds in the oil, an acid-base extraction was performed. First, the oils obtained were placed in the separation funnel along with distilled water. Then, 50 ml of 1N potassium hydroxide was added, causing p-cimenol, one of the most acidic components in the oil, to migrate to the aqueous phase.

[0098] Once the aqueous phase was isolated from the rest, it was transferred to a new separation funnel for acidification with 15 ml of 2N hydrochloric acid, and 100 ml of ether were added. In this way, the ethereal phases were obtained, which contained a higher concentration of p-cimenol.

[0099] All the ether phases were combined and filtered with anhydrous sodium sulfate as a drying agent to remove traces of water. The compound was then heated in a mafia bath to evaporate the ether and obtain the isolated phenols. This oil was absorbed with the activated and dehydrated diatomaceous earth, resulting in a fine powder product with between 14% and 16% by weight of p-cimenol relative to the total weight of the powdered extract obtained.

[0100] Example 2, Obtaining allicin from Allium sativum bulbs

[0101] To obtain the concentrated allicin extract, a solution of ethyl p-hydroxybenzoate was first prepared by diluting 0.53 g of p-hydroxybenzoate per liter of water (~3 mM). A 1 mM aqueous solution of citric acid was also prepared and used to stop the allicin formation reaction by the action of the alliinase enzyme.

[0102] Standardized garlic powder extract was macerated in an Erlenmeyer flask with the ethyl p-hydroxybenzoate solution for approximately 2.5 min and left to stand at room temperature for 30 min.

[0103] The mixture was then centrifuged at 3500 rpm for 10 minutes. Three ml of the supernatant was taken and placed in a 10 ml volumetric flask, which was then filled to the mark with 1 mM citric acid solution.

[0104] The final solution was filtered and adsorbed onto the activated, dehydrated, and ground diatomaceous earth. This resulted in an Allium sativum extract containing 13% to 14% allicin by weight relative to the total extract weight.

[0105] Example 3, Obtaining carvacrol from Origanum vulgare extract

[0106] First, the oregano plant (Origanum vulgare) was destemmed to separate the leaves from the woody parts. The Origanum vulgare leaves were ground in a porcelain mortar and dried to facilitate subsequent ethanol extraction.

[0107] 100 kilos of dry powdered Origanum vulgare leaves were macerated with 96% ethanol in a 1:3 ratio for 4 hours. The product was obtained with the consistency of an oleoresin, which must be adsorbed with silica gel and processed in a rotary evaporator at 6000 rpm for 30 minutes.

[0108] 100 kilos of powdered product yield 18 kg of carvacrol-rich extract, equivalent to between 17% and 20% of carvacrol by weight.

[0109] Example 4, Obtaining the food product of the invention Once all the active ingredients have been purified, after quantitative analysis to check the concentrations, the preparation of the final product was carried out.

[0110] This preparation took place by simple mixing of the concentrated extracts to obtain the product in solid form and the subsequent addition of the carrier, which in this case is the dehydrated, activated and ground diatomaceous earth obtained previously.

[0111] To prepare the powdered composition of the invention, 25 kg of concentrated allicin extract from Allium sativum, 15 kg of concentrated p-cimenol extract from Thymus vulgaris, and 10 kg of concentrated carvacrol extract from Origanum vulgare were added. The mixture was mixed for 10 minutes, at which point the homogeneity of the product could be ensured. The product was then ground to ensure a particle size suitable for use in industrial facilities and for the end users of the product.

[0112] Finally, 50 kg of dehydrated, activated and ground diatomaceous earth were added and the whole mixture was mixed for another 15 minutes, to obtain the final product ready to be consumed with a guaranteed minimum composition, expressed relative to the total weight of the composition, of 3.25% by weight of allicin, 2.1% by weight of p-cimenol and 1.7% by weight of carvacrol and 50% of diatomaceous earth containing 60% of diatoms of the genera Thalassiosira and Actinoptychus.

[0113] Example 5, Testing the effectiveness of the composition of the invention

[0114] To demonstrate the effectiveness of the composition of the invention on the speed of intestinal transit and the productive behavior of pigs, a test was carried out on a farm with growing and fattening pigs, where the effect of the composition of the invention on the productive parameters and the retention time of intestinal contents was verified.

[0115] This trial was conducted on a commercial farm in Arequipa with 100 weaned pigs, aged 30 to 35 days and with an initial average weight of 10.5 kg. The pigs were divided into two experimental groups and received the same rations, except for the group that included the composition of the invention. Experimental groups:

[0116] • Control group: consisting of 50 pigs receiving a standard basal diet (SBD) without additives

[0117] • Treatment group: consisting of 50 pigs receiving a DBE plus the composition of the invention at a rate of 0.5 kilograms per ton of feed continuously for 98 days

[0118] After 98 days of testing, the pigs in the treatment group receiving the composition of the invention reached an average weight of 74.43 kg, compared to 68.04 kg in the control group. This represents a 9.4% increase in the pigs' weight.

[0119] On average, pigs fed the composition of the invention had greater daily weight gain, 10.6% more, compared to pigs in the control group. The feed conversion ratio, the ratio of feed intake to daily weight gain, was 9.3% higher in the treatment group.

[0120] Intestinal transit time decreased in pigs receiving the composition of the invention; the feed was retained in the pigs for 21:33:32 minutes compared to 19:46:23 minutes for pigs in the control group. On average, pigs in the treatment group retained feed 1:47:08 minutes longer than the control pigs.

[0121] Slowing intestinal transit time had a significant positive effect on the pigs' performance parameters, resulting in improved nutrient utilization. Nutrients remained in the digestive tract longer, and because they were released slowly, they were better utilized, resulting in improved performance.

[0122] References

[0123] [1] DP Medarevic, D. Losie, and SR Ibric, “Diatoms - Nature materials with great potential for bioapplications,” Hem. Ind., vol. 70, no. 6, pp. 613-627, 2016.

[0124] [2] B. Svihus and K. Itani, “Intestinal Passage and Its Relation to Digestive Processes,” J. Appl. Poult. Res., vol. 28, no. 3, pp. 546-555, 2019. [3] IPT Nóbrega et al., “Rate of feed passage in Japanese quail,” Animal, pp. 1-8,

[0125] 2020.

[0126] [4] B. Fuerjiafu, “The effect of the grit stone on feed passage rate in broiler chickens,” Dep. Anim. Aquae. Sci., vol. Master of, pp. 1-41, 2016.

[0127] [5] M. D. Subramaniam and I. H. Kim, “Clays as dietary supplements for swine: A review,” J. Anim. Sci. BiotechnoL, vol. 6, no. 1, pp. 1-9, 2015.

[0128] [6] J. Borrell, C. Domenech, N. Martin, and A. Tesouro, “Pronutrients use in poultry nutrition (SI 1-0224),” in XXV World’s Poultry Congress, 2016, p. 97.

[0129] [7] W. Windisch, K. Schedle, C. Plitzner, and A. Kroismayr, “Use of phytogenic products as feed additives for swine and poultry,” J. Anim. Sci., vol. 86, no. 14, pp. E140-E148, 2008.

[0130] [8] A. Díaz, D. Diez, C. Domenech, and W. Ruiz, “Use of botanic molecules (Pronutrients and Cimenol Ring) for intestinal health in broilers.” International Conference on Poultry Intestinal Health (IHSIG), Rome, Italy, pp. 206-207,

[0131] 2019.

[0132] [9] J. Borrell, “Uso de pronutrientes de origen natural en veterinaria,” RACVE (Real Acad. Ciencias Vet. España), 2005.

[0133]

[0010] T. Chowdhury, S. Haidar, D. Diez, C. Domenech, and J. Pie, “Use of pronutrients (plant-based molecules) alone and combined with butyrate and probiotics to improve performance in broilers through improving gut health.” Symposium on Gut Health in Production of Food Animals, Saint Louis, Estados Unidos, p. 32,

[0134] 2021.

[0135]

[0011] M. K. Sharma, T. Dinh, and P. A. Adhikari, “Production performance, egg quality, and small intestine histomorphology of the laying hens supplemented with phytogenic feed additive,” J. AppL Poult. Res., vol. 29, no. 2, pp. 362-371,

[0136] 2020.

[0137]

[0012] J. Borrell, “Metabolites fúngicos en los alimentos,” Real Acad. Ciencias Vet. España, 1990.

[0138]

[0013] A. Marchese et al., “Update on monoterpenes as antimicrobial agents: A particular focus on p-cymene,” Materials (Basel)., vol. 10, no. 8, Aug. 2017.

[0139]

[0014] F. Tian, S. Young Woo, S. Yoo Lee, and H. Sook Chun, “p-Cymene and its derivatives exhibit antiaflatoxigenic activities against Aspergillus flavus through multiple modes of action,” AppL Biol. Chem., vol. 61.

[0140]

[0015] S. Kordali, A. Cakir, H. Ozer, R. Cakmakci, M. Kesdek, and E. Mete, “Antifungal, phytotoxic and insecticidal properties of essential oil isolated from Turkish Origanum acutidens and its three components, carvacrol, thymol and p- cymene,” Bioresour. TechnoL, vol. 99, no. 18, pp. 8788-8795, 2008.

[0141]

[0016] G. El-Saber Batiha et al., “Chemical Constituents and Pharmacological Activities of Garlic (Allium sativum L.): A Review.”

[0142]

[0017] J. Borlinghaus, F. Albrecht, M. C. H. Gruhlke, I. D. Nwachukwu, and A. J. Slusarenko, “Allicin: Chemistry and Biological Properties,” Molecules, vol. 19, pp. 12591-12618, 2014.

[0143]

[0018] A. Shang et al., “Bioactive compounds and biological functions of garlic (Allium Sativum L.),” Foods, vol. 8, no. 7, pp. 1-31, 2019.

[0144]

[0019] M. Sharifi-Rad et al, “Carvacrol and human health: A comprehensive review,” Phyther. Res., vol. 32, no. 9, pp. 1675-1687, 2018.

[0145]

[0020] K. Can Baser, “Biological and Pharmacological Activities of Carvacrol and Carvacrol Bearing Essential Oils,” Curr. Pharm. Des., vol. 14, no. 29, pp. 3106- 3119, 2008.

Claims

CLAIMS 1 Animal feed product comprising dehydrated, activated and ground diatomaceous earth, with p-cimenol, allicin and carvacrol. 2.- Animal food product according to claim 1, wherein the diatomaceous earth comprises at least 60% diatoms of the genera Thalassiosira and Actinoptychus with respect to the total amount of diatoms in the food product. 3.- Animal food product, according to claims 1 and 2, in which the dehydrated, activated and ground diatomaceous earth is present in at least 50% by weight with respect to the total weight of said food product. 4.- Animal food product according to any of the preceding claims, wherein p-cimenol is present in at least 2.1% by weight with respect to the total weight of said food product. 5.- Animal food product according to any of the preceding claims, wherein allicin is present in at least 3.25% by weight with respect to the total weight of said food product. 6.- Animal food product according to any of the preceding claims, wherein carvacrol is present in at least 1.7% by weight with respect to the total weight of said food product. 7.- Animal food product according to claim 4, comprising a 15% by weight, relative to the total weight of the food product, of a powdered extract of Thymus vulgaris in which p-cimenol is present in a percentage by weight between 14% and 16% relative to the weight of the extract. 8.- Animal food product according to claim 5, comprising a 25% by weight, relative to the total weight of the food product, of a powdered extract of Allium sativum in which allicin is present in a percentage by weight between 13% and 14% relative to the weight of the extract. 9.- Animal food product according to claim 6, comprising a 10% by weight, relative to the total weight of the food product, of a powdered extract of Origanum vulgare in which carvacrol is present in a percentage by weight between 17% and 20% relative to the weight of the extract. 10.- Method, according to any of the preceding claims, for obtaining a food product for animals comprising diatomaceous earth Dehydrated, activated and ground, with p-cimenol, allicin and carvacrol, comprising the steps: (a) obtain a diatomaceous earth with a content of at least 60% diatoms of the genera Thalassiosira and Actinoptychus with respect to the total quantity of diatoms; (b) activate and dehydrate the diatomaceous earth; (c) grinding the diatomaceous earth obtained in step (b); (d) obtaining a concentrated extract of p-cimenol from the leaves of Thymus vulgaris,' (e) obtaining an extract concentrated in allicin from the bulb of Allium sativum,' (f) obtaining a concentrated extract of carvacrol from the leaves of Origanum vulgare c, (g) mixing the diatomaceous earth from step (c) with the extracts from steps (d), (e), and (f) until homogeneous. The method according to claim 10, wherein step (a) comprises the following substeps: (al) taking at least one series of at least 10 samples each of a diatomaceous earth to be evaluated; (a2) determine by observation under optical microscope of the samples, the percentage of diatoms of the genera Thalassiosira and Actinoptychus with respect to the total quantity of diatoms; (a3) selecting a diatomaceous earth in which 100% of the observed samples comprise at least 60% diatoms of the genera Thalassiosira and Actinoptychus. Method according to claim 11, wherein in step (al), at least two series of 10 diatomaceous earth samples are taken and each sample weighs 0.9 - 1.1 grams; in step (a2), the symmetry of the frustules or silica coverings and the distribution of the pores in the diatoms are examined, determining that: i) those diatoms with radial symmetry and pores distributed homogeneously over the entire surface belong to the genus Thalassiosira; ii) those diatoms with radial symmetry and pores distributed radially belong to the genus Actinoptychus. Method according to any of claims 10 to 12, wherein the step (b) Obtaining dehydrated and activated diatomaceous earth is carried out by exposure to temperatures between 150°C and 225°C, for 25 to 30 minutes. 14.- Method according to any of claims 10 to 13, in which the step (c) comprises the following sub-stages: (the) grinding of activated dehydrated diatomaceous earth in a crushing process followed by compression, (c2) Sift the ground diatomaceous earth through a 125 pm opening sieve. 15.- Method according to claim 14, wherein in step (e) the crushing is carried out with grooved cylinders until the average size of the particles is reduced to 0.2 - 5 mm and the compression process is carried out with smooth cylinders until obtaining particles of between 50 and 150 pm where at least 95% of the particles are of a size less than 125 pm. 16.- Method according to any of claims 10 to 15, in which the step (d) comprises the following sub-stages: (di) cold press the leaves of Thymus vulgaris,' (d2) suspend the extract obtained in (di) in water; (d3) distill the oil from the leaf extract; (d4) extracting the oil from the aqueous phase; (d5) extract the phenolic content of the oil with a base and solubilize it in water; (d6) extract p-cimenol with ether; (d7) evaporate the ether. 17.- Method according to any of claims 10 to 16, in which the step (e) comprises the following sub-stages: (el) allowing allicin to be formed from alliin by the enzyme alliinase by macerating the garlic powder extract with an aqueous solution of ethyl p-hydroxybenzoate and allowing to stand at room temperature; (e2) centrifuge at 3500 rpm for at least 10 minutes; (e3) recover the supernatant and stop the enzymatic catalysis with citric acid; (e4) filter the solution obtained in (e3), and recover the filtrate obtained. 18.- Method according to any of claims 10 to 17, in which the step (f) comprises the following sub-stages: (fl) grinding and drying the leaves of Origanum vulgare to obtain a powdered product; (f2) macerate the powder obtained in (fl) in ethanol for at least 4 hours; (f3) solidify the oleoresin obtained in (f2) by absorption with silica gel and evaporate the solvent, maintaining the same temperature; (f4) centrifuge at at least 6000 rpm for at least 30 minutes; (f5) Discarding the aqueous phase. 19.- Use of the food product of any one of claims 1 to 9 for animal feed. 20.- The use according to claim 19, for feeding mammals or birds. 21.- The use according to claim 20, for feeding pigs. 22.- The use according to claim 21, wherein the food product is added to the pig feed in a proportion of 0.5 kg per ton of feed. 23.- The use according to claim 20, for feeding birds selected from the group of broiler chickens, laying hens, breeding hens, turkeys and ducks. 24.- The use according to any one of claims 19 to 23, wherein the food product is added to the animal's feed.

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