Modified attapulgite for adsorbing mycotoxins and preparation method and application thereof

Abstract

The present application belongs to the technical field of feed additives, and particularly relates to a modified attapulgite for adsorbing mycotoxins and a preparation method and application thereof. The modified attapulgite is made of attapulgite and a modifier composition, and the weight ratio of the attapulgite to the modifier composition is 100:10-17. The modifier composition is made of the following raw materials in parts by weight: zeolite 25-35 parts, shrub willow powder 5-15 parts, methionine 5-10 parts, lysine 5-10 parts, yeast cell wall 10-20 parts, chitosan oligosaccharide 3-8 parts, betaine 5-15 parts, potassium sorbate 3-8 parts, and monoglyceride fatty acid ester 5-15 parts. The modified attapulgite has multiple beneficial effects such as high-efficiency and broad-spectrum adsorption of mycotoxins, improvement of animal growth performance, regulation of immune and inflammatory responses, and the like, and has significant application value and popularization prospect in the field of feed additives.

Description

Technical Field

[0001] This invention belongs to the field of feed additive technology, specifically relating to a modified attapulgite for adsorbing mycotoxins, its preparation method, and its application. Background Technology

[0002] Aflatoxin AFB1, zearalenone (ZEA), and vomitoxin DON, among other mycotoxins, are secondary metabolites of fungi, causing contamination rates in 25% of crops globally. This results in 2% of crops losing their economic value, with annual losses exceeding hundreds of billions of US dollars. In the hot and humid regions of southern my country, the detection rate of mycotoxins in silage is as high as 70% to 100%, and 2 to 8 different toxins are often detected simultaneously in forage raw materials, exhibiting characteristics of complex contamination.

[0003] Mycotoxin contamination is a global problem affecting feed safety and animal health in livestock farming. Aflatoxin, zearalenone, and vomitoxin can accumulate in animals through the feed chain, leading to growth inhibition, organ damage, and decreased immune function. More seriously, combined mycotoxin contamination can produce synergistic toxic effects, exacerbating harm to animals. For example, combined contamination can significantly reduce feed conversion ratios, decreasing daily weight gain in finishing pigs by 15%–20%, while also affecting reproductive performance and meat quality, resulting in a substantial decline in the economic benefits of livestock farming. Furthermore, mycotoxin residues in animals not only harm animal health but also enter the human food chain through livestock products such as meat, eggs, and milk, posing a potential threat to human health.

[0004] Natural clay minerals are widely used for toxin removal due to their adsorption properties. Attapulgite, a natural layered chain-like magnesium-aluminum silicate mineral, possesses a one-dimensional nanorod-like morphology, regular nanopores, and a permanently negatively charged structure, enabling it to capture polar toxins through physical adsorption and ion exchange. However, natural attapulgite has significant limitations: poor adsorption selectivity and a rod-cluster aggregation effect. Poor adsorption selectivity manifests in the difficulty of effectively binding hydrophobic toxins on its hydrophilic surface, resulting in insufficient adsorption rates for non-polar toxins. The rod-cluster aggregation effect is reflected in the "stack-like" aggregation of rod crystals in natural minerals, leading to a low specific surface area (<100 μm²). 2 (g), insufficient exposure of surface active sites limits adsorption capacity.

[0005] Therefore, there is an urgent need to develop a new type of highly efficient attapulgite that can enhance the broad-spectrum adsorption capacity for complex mycotoxins while retaining its natural mineral safety. Summary of the Invention

[0006] The purpose of this invention is to provide a modified attapulgite for adsorbing mycotoxins, which solves the problems existing in the prior art.

[0007] The technical solution adopted in this invention is: The present invention provides a modified attapulgite for adsorbing mycotoxins, wherein the modified attapulgite is made of a composition of attapulgite and a modifier, and the weight ratio of attapulgite to the modifier composition is 100:10~17. The modifier composition is made from the following raw materials in parts by weight: 25-35 parts zeolite, 5-15 parts lemon powder, 5-10 parts methionine, 5-10 parts lysine, 10-20 parts yeast cell wall, 3-8 parts chitosan oligosaccharide, 5-15 parts betaine, 3-8 parts potassium sorbate, and 5-15 parts monoglyceride fatty acid ester.

[0008] Preferably, the weight ratio of attapulgite to modifier composition is 20:3; The modifier composition is made from the following raw materials in parts by weight: Take 30 parts of zeolite, 10 parts of lemon twig powder, 7.5 parts of methionine, 7.5 parts of lysine, 15 parts of yeast cell wall, 5 parts of chitosan oligosaccharide, 10 parts of betaine, 5 parts of potassium sorbate, and 10 parts of monoglyceride fatty acid ester.

[0009] A second aspect of the present invention provides a method for preparing the modified attapulgite, comprising the following steps: After attapulgite is crushed, sulfuric acid solution is added and aged to obtain activated attapulgite. Zeolite, Ligusticum chuanxiong powder, methionine, lysine, yeast cell wall, chitosan oligosaccharide, betaine, potassium sorbate and monoglyceride fatty acid ester are mixed in the stated weight parts to obtain a modifier composition; The activated attapulgite and the modifier composition are mixed, filtered, and dried to obtain the modified attapulgite.

[0010] Preferably, the aging conditions are at room temperature for 10 to 16 hours.

[0011] Preferably, the filtration conditions are: 0.6MPa~0.8MPa.

[0012] Preferably, the drying conditions are 80℃~110℃.

[0013] A third aspect of the present invention provides an application of the modified attapulgite, wherein the modified attapulgite is used to prepare an additive for animal feed to adsorb mycotoxins in animal feed, thereby improving the production performance of animals.

[0014] Preferably, the mycotoxin includes at least one of aflatoxin, zearalenone, and vomitoxin.

[0015] Preferably, the aflatoxin includes at least one of AFB1, AFB2, AFG1 and AFM1.

[0016] Preferably, improving animal production performance includes at least one of the following: 1) Increase the animal's weight; 2) Increase the average daily weight gain of animals; 3) Increase the animals' average daily feed intake; 4) Reduce the feed conversion ratio of animals.

[0017] Compared with the prior art, the beneficial effects of the present invention are: This invention provides a modified attapulgite for adsorbing mycotoxins. The modified attapulgite is made from a composition of attapulgite and a modifier, with a weight ratio of attapulgite to the modifier composition of 100:10-17. The modifier composition is made from the following raw materials in parts by weight: 25-35 parts zeolite, 5-15 parts Ligusticum chuanxiong powder, 5-10 parts methionine, 5-10 parts lysine, 10-20 parts yeast cell wall, 3-8 parts chitosan oligosaccharide, 5-15 parts betaine, 3-8 parts potassium sorbate, and 5-15 parts monoglyceride fatty acid ester. The modified attapulgite of this invention, through synergistic compounding of attapulgite with a specific modifier composition (containing zeolite, Ligusticum chuanxiong powder, amino acids, yeast cell wall, chitosan oligosaccharide, betaine, potassium sorbate, and monoglyceride fatty acid ester), significantly enhances the broad-spectrum adsorption capacity for various mycotoxins such as aflatoxin B1, vomitoxin, and zearalenone. UPLC-QQQ-MS analysis confirmed that this modified attapulgite significantly reduced toxin concentrations in moldy feed. Animal studies further demonstrated that it effectively alleviated mycotoxin-induced growth inhibition in Hu sheep, significantly increasing final body weight, average daily weight gain, and average daily feed intake, while reducing the feed conversion ratio. Simultaneously, it restored levels of immunoglobulins (IgG, IgA, IgM) and inflammatory factors (TNF-α, IL-6, IL-10), alleviating immune stress and inflammatory damage. This invention overcomes the shortcomings of poor adsorption selectivity and rod-crystal aggregation of natural attapulgite through a triple dynamic synergistic mechanism of "physical adsorption - intestinal barrier strengthening - in vivo metabolic regulation," providing a highly efficient, safe, and green feed additive solution for the control of complex mycotoxin contamination, with promising application prospects and economic benefits. Attached Figure Description

[0018] Figure 1 The effects of different treatments on the content of major mycotoxins in feed.

[0019] Figure 2 The effects of different treatments on the growth performance of Hu sheep are shown in Figure 1. A: Initial body weight; B: Final body weight; C: Average daily weight gain; D: Average daily feed intake; E: Feed conversion ratio. Different letters indicate significant differences between groups (P < 0.05).

[0020] Figure 3The effect of attapulgite treatment on immunoglobulin and inflammatory factor levels in Hu sheep contaminated with mycotoxins. A: IgG level; B: IgA level; C: IgM level; D: TNF-α level; E: IL-6 level; F: IL-10 level. Different letters indicate significant differences between groups (P<0.05). Data are expressed as mean ± standard error. Detailed Implementation

[0021] The present invention will be further illustrated below with specific embodiments, but these embodiments do not limit the scope of the invention. Modifications or substitutions to the details and form of the technical solutions of the present invention may be made without departing from the spirit and scope of the invention, but all such modifications or substitutions fall within the protection scope of the present invention.

[0022] The inventive concept of this invention is as follows: The advanced nature of the modified attapulgite prepared by this invention lies in its triple dynamic action mechanism of "adsorption, degradation and repair".

[0023] The first layer (in the intestine): Attapulgite, with its nanoscale zeolite channels (0.37nm × 0.64nm), adsorbs mycotoxins (such as aflatoxin B1 and zearalenone). Simultaneously, the abundant silanol and aluminol groups on its surface form hydrogen bonds and coordination complexes with the β-carbonyl lactone rings of the toxin molecules. Zeolite, through its three-dimensional cage structure and π-π stacking, efficiently captures mycotoxins (such as zearalenone and ochratoxin A). Together, they achieve full-spectrum physical adsorption of toxins, from polar to weakly polar, forming stable complexes that are excreted with feces, completing the "mycotoxin removal" process before the toxins enter the intestinal epithelial cells.

[0024] The second layer (intestinal wall): Mannooligosaccharides in the yeast cell wall directly bind to toxin molecules such as zearalenone and ochratoxin A through hydrogen bonds and hydrophobic interactions, reducing the concentration of free toxins in the intestine, thereby reducing the chance of toxins binding to glucose transporters and organic anion transport peptides on the surface of intestinal epithelial cells, and indirectly inhibiting the transmembrane absorption of toxins; Chitosan oligosaccharides upregulate the expression of tight junction proteins such as Claudin-1, Occludin, and ZO-1 by activating the protein kinase C signaling pathway, thus strengthening the intestinal physical barrier; Monoglycerides of fatty acids insert into the phospholipid bilayer of harmful bacteria, disrupting cell membrane integrity and inhibiting opportunistic pathogens such as Clostridium perfringens; Water-soluble polysaccharides in Caragana korshinskii powder act as prebiotics to promote the proliferation of Lactobacillus and Bifidobacterium, and its crude fiber and crude protein content can provide certain nutritional support; Betaine acts as an osmotic regulator to maintain the volume homeostasis of intestinal epithelial cells and removes reactive oxygen species by enhancing the activity of glutathione peroxidase and superoxide dismutase. These components form a multi-layered defense between toxins and the body, including "blocking binding, strengthening the barrier, regulating the microbiome, and protecting cells."

[0025] The third aspect (in vivo metabolism): Flavonoids in *Leonurus japonicus* powder can regulate the activity of hepatic drug-metabolizing enzymes. They affect the expression of cytochrome P450 enzyme systems (CYP1A1, CYP3A4) through the aryl hydrocarbon receptor (AhR) signaling pathway. While enhancing phase I toxin metabolism, attention should be paid to the balance of metabolic activation products. Methionine generates cysteine ​​through the methionine cycle and transsulfurization pathway, providing the rate-limiting precursor for glutathione (GSH) synthesis. Sufficient GSH serves as a substrate in the binding reaction catalyzed by glutathione-S-transferase (GST), converting the activated toxin intermediate into water-soluble, non-toxic metabolites that are excreted through bile and urine. Betaine, as a methyl donor, participates in the methionine cycle, indirectly supporting GSH synthesis and metabolism. At the same time, it enhances the activity of glutathione peroxidase (GPx) and superoxide dismutase (SOD), systematically scavenging reactive oxygen species and reducing malondialdehyde levels, protecting the liver and kidneys from oxidative damage.

[0026] Fourthly, chitosan oligosaccharides insert into the interlayer of attapulgite through electrostatic self-assembly to open up the pores, and synergistically construct a three-dimensional hydrogen bond network with yeast polysaccharides to enhance chemical adsorption; monoglyceride fatty acid esters form hydrophobic microregions on the mineral surface, and capture weakly polar toxins through hydrophobic distribution and π-π stacking cascade; Ligusticum chuanxiong powder polysaccharides provide steric hindrance to stabilize colloidal dispersion and prevent agglomeration and pore blockage; betaine synergistically regulates surface charge and pH microenvironment.

[0027] The above four interactions form a synergistic system of "interfacial hybridization-hydrophobic partitioning-colloidal stability", which simultaneously improves the adsorption site density, adsorption energy and kinetic rate, and achieves efficient capture of toxins of the entire spectrum.

[0028] To enable those skilled in the art to better understand and implement the technical solutions of this invention, the invention will be further described below with reference to specific embodiments. Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art.

[0029] The raw materials used in this invention are all obtained through commercial channels, specifically including: Zeolite: The zeolite used in this invention is selected from Aladdin's 4A zeolite molecular sieve, product number F109287; it can also be selected from feed-grade clinoptilolite (such as Hebei Mufeng feed-grade zeolite powder, 200 mesh).

[0030] Caragana powder (160 mesh): belongs to the Caragana genus ( Caragana korshinskii The dried branch powder is made from fresh lemon branches that have been dried at 105℃ and mechanically pulverized through a 160-mesh standard sieve.

[0031] Methionine: Selected from Ningxia Ziguang Tianhua Methionine Co., Ltd. DL-methionine (feed grade, purity ≥98.5%, CAS 59-51-8).

[0032] Lysine: Selected from CJ brand L-lysine hydrochloride (feed grade, purity ≥98.5%, CAS 67-27-2) of CJ Biotechnology Co., Ltd.

[0033] Yeast cell wall: Selected from Angel Yeast Co., Ltd.'s "Angel" brand yeast cell wall polysaccharide (product number YCW-20, mannan oligosaccharide ≥20%, β-glucan ≥20%).

[0034] Chitosan oligosaccharide: Selected from "Honghai" brand chitosan oligosaccharide (product number HH-COS-3K, molecular weight ≤3000 Da, degree of deacetylation ≥90%, CAS 148411-57-8) from Qingdao Honghai Biotechnology Co., Ltd.

[0035] Betaine: Selected from "Xiangweisi" brand betaine hydrochloride (anhydrous betaine, purity ≥98%, CAS 590-46-5) of Shandong Xiangweisi Biotechnology Co., Ltd.

[0036] Potassium sorbate: Selected from "Wanglong" brand potassium sorbate (food grade / feed grade, purity ≥98%, CAS 590-00-1) of Shandong Wanglong Technology Co., Ltd.

[0037] Monoglyceride fatty acid esters: Selected from "Monoglycerides" (HLB 3.8, monoglyceride content ≥40%, CAS 31566-31-1) of Henan Zhengtong Food Technology Co., Ltd.

[0038] The specific goods or equivalent commercially available products mentioned above can be used to implement this invention, therefore there are no specific restrictions on their sources.

[0039] In the following examples, 1g represents one part by weight.

[0040] Example 1 A modified attapulgite for adsorbing mycotoxins is prepared by the following method: S1. Preparation of activated attapulgite.

[0041] The natural attapulgite clay mineral is crushed and passed through a 200-mesh sieve. Then, sulfuric acid aqueous solution is sprayed into it to dissolve the carbonates in the mineral. The mineral is then aged at room temperature for about 12 hours to fully activate it and obtain activated attapulgite.

[0042] In this step, the total amount of sulfuric acid sprayed is 1% of the natural attapulgite clay mineral content, and the water content of the clay mineral is 8%. Therefore, there is no longer a limit on the concentration of the sulfuric acid aqueous solution here.

[0043] S2. Preparation of the modifier composition: Weigh each raw material according to the following weights and mix them to obtain the modifier composition.

[0044] Take 30g of zeolite, 10g of lemon powder, 7.5g of methionine, 7.5g of lysine, 15g of yeast cell wall, 5g of chitosan oligosaccharide, 10g of betaine, 5g of potassium sorbate, and 10g of monoglyceride fatty acid ester.

[0045] S3. The activated attapulgite is mixed with the modifier composition.

[0046] Take 100g of activated attapulgite and 15g of the modifier composition, mix them thoroughly in a ball mill, and then add an aqueous solution containing 10% ethanol to fully wet the mixture. Continue mixing under ball milling conditions and allow the reaction to occur under mechanical force. This process involves the transformation of inert Si-O-Si bonds on the mineral surface into active Si-OH bonds under mechanical ball milling. These Si-OH bonds can form strong hydrogen bonds with small molecules such as amino acids, betaine, and potassium sorbate, locking them onto the mineral surface. Ethanol can replace water in the attapulgite structure, while monoglycerides of fatty acids can adhere to the mineral surface to form hydrophobic microdomains, which is beneficial for the adsorption of weakly polar mycotoxins. To ensure thorough mixing and complete reaction with minimal solvent usage, the ethanol aqueous solution is added until the slurry is still flowable. Then, plate and frame filtration is performed to obtain a filter cake at a filtration pressure of 0.7 MPa. The cake is then dried and pulverized at 110℃ to obtain a 200-mesh solid powder, which is the modified attapulgite feed additive product.

[0047] Example 2 An application of modified attapulgite for adsorbing mycotoxins is as follows: 1. Experimental protocol.

[0048] (1) Experimental animals and design.

[0049] Forty 8-month-old Hu sheep with similar genetic backgrounds and ages were selected for this experiment, with an initial weight of 32.12 ± 1.70 kg. Before the experiment, all sheep were dewormed and vaccinated with a triple-quadruple vaccine. The sheep were randomly divided into four treatments according to weight, with 10 sheep in each treatment. The composition and nutritional levels of the feed for each group are shown in Tables 1 and 2. The specific treatment groups and their feeds are as follows:

[0050] The basal diet group (CG); fed the basal diet group; Mycotoxin group (MG): Replacing a portion of the normal corn in CG with moldy corn; Mycotoxin + Unmodified Attapulgite Group (MNAG): 2% activated attapulgite (a product of S1) was added to the MG diet. Mycotoxin + Modified Attapulgite Group (MMAG): 2% of the modified attapulgite prepared in Example 1 was added to the MG diet.

[0051] Before the formal trial began, blood was collected from the experimental sheep, and serum was separated. Brucellosis was detected using the rose bengal plate method to ensure no infection. After confirming the absence of infection, the sheep underwent a 10-day acclimatization period to ensure adaptation to the feed and environment. The formal trial lasted 90 days, using a pen-based feeding system with free access to roughage and water. Roughage was fed daily at 6:30 AM, ensuring that more than 5% roughage remained in the feed troughs before concentrate feeding. The weight of remaining roughage was recorded, and the troughs were cleaned and concentrate fed at 5:30 PM. After the trial, the experimental sheep were slaughtered, and their production performance and meat quality were measured.

[0052] Table 1 Feed Trial Formulations Table 2. Nutritional composition of diets for each treatment group Note: In Table 1, " / " indicates that this item is not present.

[0053] (2) Production performance measurement.

[0054] During the formal trial, the amount of leftover feed for each group of sheep was recorded daily to accurately calculate their feed intake. The sheep were weighed on days 0, 30, 60, and 90 of the trial to obtain body weight data. Production performance was calculated using this data, using the following formula:

[0055] 1) Average daily gain (ADG): The average weight gain per day during the trial period, calculated using the following formula: .

[0056] in, A Average daily weight gain (kg / d); B Final weight (kg); C Initial weight (kg); d Experimental days (d).

[0057] 2) Average daily feed intake (ADFI): The average amount of feed consumed per day during the experiment, calculated using the following formula: .

[0058] in, a Average daily feed intake (g / d); b Feed consumption (g); d Experimental days (d).

[0059] 3) Feed conversion ratio: The ratio of average daily feed intake to average daily weight gain, calculated using the following formula; .

[0060] in, D Material weight ratio; a Average daily feed intake (g / d); F Average daily weight gain (g).

[0061] (3) The content of mycotoxins in the quantitative diet.

[0062] The contents of aflatoxin B1, vomitoxin and zearalenone in the diet were quantitatively analyzed using UPLC-QQQ-MS.

[0063] (4) Slaughtering performance.

[0064] After the experiment, all Hu sheep were fasted for 24 hours before slaughter. All slaughter procedures were conducted under ethically permissible conditions and strictly adhered to the Regulations on the Management of Laboratory Animals and international ethical guidelines. Pre-slaughter live weight was measured, and the neck was slit for bleeding. After bleeding, the sheep's skin, head, hooves, and internal organs were removed, and carcass weight was measured, along with the weight of the internal organs. Dressing percentage, organ index, GR value, and carcass weight were calculated. Additionally, the tissue thickness located between the 12th and 13th ribs, 11 cm from the midline of the spine, was measured as the carcass fat content (GR value). The longissimus dorsi muscle was collected 45 minutes and 24 hours post-slaughter, and the pH was measured using a portable pH meter. All samples were rapidly frozen in liquid nitrogen and then stored at -80°C for subsequent analysis. The organ index is an important indicator reflecting the growth and development of animal organs and their overall health.

[0065] (5) Physiological and biochemical indicators.

[0066] After collecting sheep serum samples, the concentrations of creatinine (CREA), gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were measured according to the kit instructions. In addition, the concentrations of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) were also measured.

[0067] The specific operating procedure is as follows: First, take out the kit and the sample to be tested, and let them stand at room temperature for 1 hour. Then, add the sample and control to the microplate and incubate with the antibody working solution. After incubation, add the enzyme label, wash the plate, and perform a second incubation. Then, add the substrate solution, terminate the reaction, and measure the absorbance at the specified wavelength. Finally, determine the values ​​of each index according to the standard curve.

[0068] (6) Meat quality indicators.

[0069] 1) Flesh color: The light absorbance of the longissimus dorsi muscle was measured using a colorimeter, and the brightness (L*), redness (a*), and yellowness (b*) values ​​of the meat samples were recorded. To ensure the reliability of the data, each meat sample was measured in triplicate, with three different points selected for each parallel measurement.

[0070] pH value: pH values ​​were measured at 45 min and 24 h post-slaughter. The electrodes of a portable pH meter were inserted approximately 2 cm into the longissimus dorsi muscle of the sheep. The readings were repeated three times, and the average value was calculated to ensure the reliability of the data.

[0071] 2) Shear Force: The longissimus dorsi muscle was harvested, surface fat and connective tissue were removed, and the muscle samples were trimmed to approximately 6cm × 3cm × 3cm. The muscle samples were placed in a No. 6 resealable bag and then heated in an 80°C water bath until the center temperature reached 70°C. Afterward, the muscle samples were hung in a cool, ventilated, and dry environment and air-dried at room temperature (approximately 20°C) for 20 minutes. Using a 1.27cm diameter circular sampler, samples were taken from the center of the meat sample along the muscle fiber direction and cut into 1.5cm × 1.0cm × 1.0cm samples, ensuring at least three samples were taken. Subsequently, the shear force of the meat samples was measured using a texture analyzer. The results are expressed in Newtons (N), and the average value was calculated.

[0072] 3) Cooking Loss: Remove fat and connective tissue from the surface of the longissimus dorsi muscle and trim it into muscle samples approximately 6cm × 3cm × 3cm. Place the muscle samples in a No. 5 food storage bag, squeeze out the gas, seal the bag, and place it in an 80℃ constant temperature water bath for 1 hour. Remove the muscle samples and hang them in a cool, dry place at room temperature for 20 minutes. First, absorb the surface moisture of the muscle samples with filter paper, then use a balance with an accuracy of 0.001 grams to measure the weight of the samples before and after cooking. The calculation formula is:

[0073] .

[0074] in, e : Loss during cooking; f Weight before cooking, kg; g Weight after steaming or boiling, kg.

[0075] 4) Drip Loss: Remove the longissimus dorsi muscle from the surface fat and connective tissue, and trim it along the muscle fibers to a length of 2cm × 1cm × 1cm. Hang one end of the meat sample inside a disposable paper cup using an S-hook, ensuring the sample does not touch the cup wall when stationary. Place the paper cup in a No. 5 resealable bag, leaving the upper end of the S-hook protruding from the bag opening. Seal the bag and store it in a 4℃ refrigerator for 24 hours. Remove the meat and gently wipe away surface moisture with filter paper. Measure the weight of the meat before and after hanging using a balance with an accuracy of 0.001 grams, and calculate the drip loss using the following formula:

[0076] .

[0077] in, h Drip loss; i : Hanging weight, g; j : after hanging, g.

[0078] 5) Moisture content, denoted as k A portion of the longissimus dorsi muscle was taken and weighed, and recorded as follows: m The unit is g; freeze-dry using a freeze dryer until the weight no longer decreases, and record the weight as g. n The unit is g.

[0079] .

[0080] 2. Experimental results.

[0081] (1) UPLC-QQQ-MS method was used to quantify the mycotoxin content in the diets of each treatment group. The UPLC-QQQ-MS results showed that replacing normal corn in the diet with moldy corn significantly increased the content of aflatoxin B1 (AFB1), vomitoxin (DON), and zearalenone (ZEA) in the diet. In the MG group, the AFB1 concentration was 4.5 ppb, which was about 5 times that of the CG group; the DON concentration was 767.1 ppb, which was nearly 2 times that of the CG group; and the ZEA concentration was 750.3 ppb, which was 7 to 8 times that of the CG group. This indicates that mycotoxin contamination poses a serious threat to the safety of the diet and verifies the success of the model. The results are shown in […]. Figure 1 .

[0082] By adding activated attapulgite (MNAG group) and modified attapulgite (MMAG group) to the diets of the model groups, it was found that both treatments reduced the mycotoxin content in the diets, with AFB1 concentrations decreasing to 4.2 ppb and 3.5 ppb, respectively (MNAG group and MMAG group). Modified attapulgite showed a better effect in reducing ZEA concentration (669.3 ppb in MMAG group). These results indicate that modified attapulgite has a stronger adsorption capacity in mitigating mycotoxin pollution, providing important theoretical basis and practical guidance for improving diet safety and controlling mycotoxin pollution.

[0083] (2) The addition of modified attapulgite improved the decline in meat sheep production performance caused by mycotoxins.

[0084] Mycotoxin contamination significantly reduced the final body weight, average daily weight gain, and average daily feed intake of Hu sheep, while significantly increasing the feed conversion ratio, indicating that mycotoxins have a significant negative impact on the growth performance and feed utilization efficiency of Hu sheep (P<0.05). Compared with the mycotoxin contaminated group (MG group), the activated attapulgite treatment group (MNAG group) and the modified attapulgite treatment group (MMAG group) significantly improved the final body weight, average daily weight gain, and average daily feed intake of Hu sheep, and significantly reduced the feed conversion ratio (P<0.05). Among them, modified attapulgite performed better in improving the feed conversion ratio. Figure 2 The results showed that modified attapulgite is an effective mycotoxin adsorbent, which can alleviate mycotoxin contamination of feed, significantly improve feed utilization efficiency and the production performance of Hu sheep, and provide important practical evidence for improving feed safety.

[0085] (3) Modified attapulgite alleviates the effects of mycotoxins on immune indicators and inflammatory factors in Hu sheep.

[0086] This invention investigated the effects of mycotoxin contamination (MG group), activated attapulgite treatment (MNAG group), modified attapulgite treatment (MMAG group), and basal diet (CG group) on immune indicators and inflammatory factors in Hu sheep. The results are as follows: Figure 3 The results showed that mycotoxin contamination significantly increased the levels of immunoglobulins (IgG, IgA, IgM) in Hu sheep, while also significantly increasing the expression of inflammatory factors (TNF-α, IL-6) and inhibiting the expression of anti-inflammatory factor (IL-10) (P<0.05), indicating that mycotoxins have a serious negative impact on the immune system and inflammatory response of Hu sheep.

[0087] Specifically, the levels of IgG, IgA, and IgM in the MG group were significantly higher than those in the CG group (P<0.05), suggesting that mycotoxins induced an immune stress response. Both the MNAG and MMAG groups significantly reduced these immunoglobulin levels (P<0.05), demonstrating the alleviating effect of attapulgite treatment on mycotoxin toxicity, with the MMAG group showing a more significant regulatory effect. Regarding inflammatory factors, the levels of TNF-α and IL-6 in the MG group were significantly increased (P<0.05), indicating that mycotoxins induced an inflammatory response. The MNAG and MMAG groups significantly reduced TNF-α and IL-6 levels (P<0.05), with the MMAG group showing a better effect than the MNAG group. Furthermore, the IL-10 level in the MG group was significantly lower than that in the CG group (P<0.05), suggesting that mycotoxins inhibited the expression of anti-inflammatory factors. The MNAG and MMAG groups significantly increased IL-10 levels (P<0.05), with the MNAG group showing a more pronounced effect. In summary, mycotoxin contamination has a significant toxic effect on the immune system and inflammatory factors of Hu sheep. Natural attapulgite and modified attapulgite can effectively alleviate these negative effects, improve the immune status and anti-inflammatory capacity of Hu sheep, and provide important theoretical basis and practical guidance for the prevention and control of mycotoxin contamination and the application of attapulgite.

[0088] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0089] The embodiments described above are merely examples of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. A modified attapulgite for adsorbing mycotoxins, characterized in that, The modified attapulgite is made from a composition of attapulgite and a modifier, wherein the weight ratio of attapulgite to the modifier composition is 100:10~17. The modifier composition is made from the following raw materials in parts by weight: 25-35 parts zeolite, 5-15 parts lemon powder, 5-10 parts methionine, 5-10 parts lysine, 10-20 parts yeast cell wall, 3-8 parts chitosan oligosaccharide, 5-15 parts betaine, 3-8 parts potassium sorbate, and 5-15 parts monoglyceride fatty acid ester.

2. The modified attapulgite as described in claim 1, characterized in that, The weight ratio of attapulgite to modifier composition is 20:3; The modifier composition is made from the following raw materials in parts by weight: Take 30 parts of zeolite, 10 parts of lemon twig powder, 7.5 parts of methionine, 7.5 parts of lysine, 15 parts of yeast cell wall, 5 parts of chitosan oligosaccharide, 10 parts of betaine, 5 parts of potassium sorbate, and 10 parts of monoglyceride fatty acid ester.

3. The method for preparing modified attapulgite as described in claim 1, characterized in that, The steps are as follows: After attapulgite is crushed, sulfuric acid solution is added and aged to obtain activated attapulgite. Zeolite, Ligusticum chuanxiong powder, methionine, lysine, yeast cell wall, chitosan oligosaccharide, betaine, potassium sorbate and monoglyceride fatty acid ester are mixed in the stated weight parts to obtain a modifier composition; The activated attapulgite and the modifier composition are mixed, filtered, and dried to obtain the modified attapulgite.

4. The preparation method according to claim 3, characterized in that, The aging conditions are room temperature and 10-16 hours.

5. The preparation method according to claim 3, characterized in that, The filtration conditions are: 0.6MPa~0.8MPa.

6. The preparation method according to claim 3, characterized in that, The drying conditions are: 80℃~110℃.

7. The application of the modified attapulgite as described in claim 1, characterized in that, The modified attapulgite is used to prepare an additive for animal feed to adsorb mycotoxins in animal feed, thereby improving animal production performance.

8. The application as described in claim 7, characterized in that, The mycotoxins include at least one of aflatoxin, zearalenone, and vomitoxin.

9. The application as described in claim 8, characterized in that, The aflatoxins include at least one of AFB1, AFB2, AFG1, and AFM1.

10. The application as described in claim 7, characterized in that, Improving animal production performance includes at least one of the following: 1) Increase the animal's weight; 2) Increase the average daily weight gain of animals; 3) Increase the animals' average daily feed intake; 4) Reduce the feed conversion ratio of animals.

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