A mycotoxin toxicity mitigant and uses thereof

Abstract

The application discloses a mycotoxin toxicity reducing agent and application, and the composition comprises peanut shell extract 30-40 parts by weight, naringin 2-5 parts by weight, and pinene-4-ol 1-3 parts by weight. In mycotoxin contaminated feed, the peanut shell extract, the naringin and the pinene-4-ol cannot relieve the toxicity of the mycotoxin alone. Compared with the combination of any one of the three components, the complete addition of the three components can synergistically relieve the mitochondrial calcium overload induced by the mycotoxin, significantly relieve the mitochondrial damage caused by the mycotoxin and improve the mitochondrial function, and significantly relieve the toxicity effect of the mycotoxin on livestock and poultry. The three components have significant interaction in relieving the mitochondrial calcium overload and improving the mitochondrial function, and have a positive synergistic combination effect. Through the synergistic effect of the components, the application can significantly relieve the toxicity effect of the mycotoxin on livestock and poultry, and improve the production performance and health status of the livestock and poultry.

Description

Technical Field

[0001] This invention relates to the field of additives for livestock and poultry farming, specifically to a mycotoxin toxicity reducer and its application. Background Technology

[0002] Feed mold and mycotoxin contamination cause significant economic losses to livestock and poultry farming. Aflatoxin, vomitoxin (deoxynivalenol), zearalenone, ochratoxin, fumonisin, and T-2 toxin are common mycotoxins in feed. When multiple mycotoxins are present simultaneously, their toxic effects are not simply additive, but rather exhibit a significant synergistic enhancement. Even if the mycotoxin contamination level in feed is within the relevant feed hygiene standards, the synergistic effect of multiple toxins still harms livestock and poultry health and production performance.

[0003] Adsorption and degradation are the two main mechanisms for reducing the toxicity of mycotoxins in feed. However, most adsorbents can only adsorb highly polar mycotoxins such as aflatoxin, and their adsorption effect on toxins such as vomitoxin is poor. Problems with the biodegradation of mycotoxins include: the detoxifying strains are prone to degradation and their degradation activity is unstable; the detoxifying enzymes have poor stability, and because degrading enzymes are highly specific, and feed contains multiple mycotoxins, a single degrading enzyme cannot comprehensively cover them, thus requiring the formulation of multiple enzymes.

[0004] Mitochondria are the target of mycotoxins. Regardless of the complexity of the types and sources of mycotoxins, one of their core toxic mechanisms is the disruption of mitochondrial function. Mycotoxin attenuation technologies that protect mitochondria are a universal strategy to intercept toxic pathways at the "end point," and can address the synergistic attack of multiple mixed toxins. Mycotoxins induce mitochondrial calcium overload, leading to mitochondrial dysfunction, which is an important pathway of mycotoxin toxicity and can serve as a target for mitigating mycotoxin toxicity.

[0005] Therefore, developing a mycotoxin attenuator that mitigates mycotoxin toxicity at the mitochondrial level would be of great significance for livestock and poultry farming. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of conventional adsorption and degradation technologies in reducing mycotoxins, and to provide a mycotoxin toxicity reducer and its application.

[0007] To achieve the above-mentioned objectives, the technical solution adopted by this invention is as follows:

[0008] In a first aspect, the present invention provides a mycotoxin toxicity reducer comprising the following components in parts by weight: 30-40 parts of peanut shell extract, 2-5 parts of rutin, and 1-3 parts of terpinene-4-ol.

[0009] As a preferred embodiment of the first aspect above, the mycotoxin toxicity reducing agent, by weight, is composed of 30-40 parts of peanut shell extract, 2-5 parts of rutin, and 1-3 parts of terpinene-4-ol.

[0010] As a preferred embodiment of the first aspect above, the peanut shell extract is obtained from peanut shells by ethanol extraction. The extraction method is as follows: after pulverizing dried peanut shells, ethanol is used as a solvent to extract the peanut shells multiple times at a temperature of 40~60℃. The extracts are combined, concentrated, purified and dried to obtain the peanut shell extract.

[0011] Furthermore, during the extraction process using ethanol as a solvent, an extraction aid is required, including polyvinylpyrrolidone or polyethylene glycol 200; the purification operation employs alcohol precipitation or water washing.

[0012] As a preferred embodiment of the first aspect above, the peanut shell extract is obtained from peanut shells by ethanol extraction.

[0013] In a second aspect, the present invention provides a feed containing a mycotoxin toxicity reducer as described in any one of the first aspects above.

[0014] As a preferred embodiment of the second aspect above, the amount of mycotoxin toxicity reducer added to the feed is 0.015 wt.% to 0.03 wt.%.

[0015] As a preferred embodiment of the second aspect above, the feed is pig feed or broiler feed.

[0016] Thirdly, the present invention provides an application of the mycotoxin toxicity reducer as described in any of the first aspects above in livestock and poultry feeding, specifically used to alleviate the toxic effects of mycotoxins by regulating mitochondrial calcium transporters, alleviating mitochondrial calcium overload and improving mitochondrial function.

[0017] As a preferred embodiment of the third aspect above, the mycotoxin toxicity reducer is added to the feed for livestock and poultry in the form of an additive.

[0018] As a preferred embodiment of the third aspect above, the livestock or poultry is pig or broiler chicken.

[0019] The mycotoxin toxicity reducing agent of the present invention has the following characteristics:

[0020] (1) This invention has developed a mycotoxin toxicity reducer by means of network pharmacology, high-throughput screening, molecular docking and other technologies. It can significantly affect the expression of mitochondrial calcium uniporter (MCU) protein and alleviate mitochondrial calcium overload, enhance mitochondrial oxidative phosphorylation, improve mitochondrial function, and thus alleviate the toxic effects of mycotoxins.

[0021] (2) In mycotoxin-contaminated feed, the addition of peanut shell extract, rutin, and terpinene-4-ol alone could not alleviate the toxicity of mycotoxins. Compared with the combination of two components (i.e., the combination lacking one of the components), the addition of the three components of this invention can synergistically alleviate mycotoxin-induced mitochondrial calcium overload, significantly alleviate mycotoxin-induced mitochondrial damage and improve mitochondrial function, and significantly alleviate the toxic effects of mycotoxins on livestock and poultry.

[0022] (3) Peanut shell extract, rutin, and terpinene-4-ol have significant interaction effects in alleviating mitochondrial calcium overload and improving mitochondrial function, and have a positive significant synergistic effect.

[0023] (4) The mycotoxin toxicity reducer of the present invention can significantly alleviate the toxic effects of mycotoxins in feed on livestock and poultry through the synergistic effect of each component, and improve the production performance and health status of livestock and poultry. Attached Figure Description

[0024] Figure 1 The values ​​represent the growth performance, intestinal barrier, liver function, oxidative stress, and inflammation of pigs in each group in Example 1. Different letters indicate significant differences (P<0.05).

[0025] Figure 2 The values ​​represent MCU, mitochondrial calcium, and mitochondrial function in each group of Example 1, with significant differences observed for groups with different letters (P<0.05). The mean value for the control group was 1.00, and the values ​​for other groups represent relative fold increases.

[0026] Figure 3 The values ​​represent the growth performance, intestinal barrier, liver function, oxidative stress, and inflammation of pigs in each group in Example 2. Different letters indicate significant differences (P<0.05).

[0027] Figure 4 The values ​​for MCU, mitochondrial calcium, and mitochondrial function in each group in Example 2 are given, with significant differences observed for groups with different letters (P<0.05). The mean value for the control group is 1.00, and the values ​​for other groups represent relative fold increases.

[0028] Figure 5 The values ​​represent the growth performance, intestinal barrier, liver function, oxidative stress, and inflammation of broilers in Example 3, with different letters indicating significant differences (P<0.05).

[0029] Figure 6 The values ​​represent MCU, mitochondrial calcium, and mitochondrial function in each group in Example 3, with significant differences observed for groups with different letters (P<0.05). The mean value for the control group was 1.00, and the values ​​for other groups represent relative folds. Detailed Implementation

[0030] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention can be practiced in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below. Technical features in various embodiments of the present invention can be combined accordingly without mutual conflict.

[0031] This invention, through studying the molecular mechanisms of mycotoxin toxicity, discovered that mycotoxins damage mitochondria and impair mitochondrial function, weakening mitochondrial oxidative phosphorylation and ATP production. Abnormal function of the mitochondrial calcium uniporter (MCU) and mitochondrial calcium overload are important mechanisms by which mycotoxins damage mitochondria and can serve as targets for mitigating mycotoxin toxicity. Based on this, this invention provides a mycotoxin toxicity reducer, comprising at least three components in parts by weight: 30-40 parts by weight of peanut shell extract, 2-5 parts by weight of rutin, and 1-3 parts by weight of terpinene-4-ol.

[0032] The peanut shell extract described above is obtained from peanut shells using an ethanol extraction method. This type of peanut shell extract can be prepared using existing methods or purchased commercially. In an embodiment of the present invention, the ethanol extraction method for peanut shell extract is as follows: dried peanut shells are pulverized, and ethanol is used as a solvent for multiple extractions at a temperature of 40-60°C (an extraction aid, such as polyvinylpyrrolidone or polyethylene glycol 200, may be added during the extraction process). The extracts are combined, concentrated, purified (using methods such as alcohol precipitation or water washing), and dried to obtain the peanut shell extract. Both rutin and terpinene-4-ol mentioned above can be purchased commercially. The CAS number for rutin is 14259-46-2, and the CAS number for terpinene-4-ol is 562-74-3.

[0033] It should be noted that the above three components are the core components of this mycotoxin toxicity reducer. In theory, in addition to the above three components, other auxiliary components can be added to this mycotoxin toxicity reducer, and there are no restrictions on this.

[0034] The mycotoxin toxicity reducer of the present invention can be added to a basic feed to form a feed with mycotoxin toxicity reduction function. The recommended addition amount of the mycotoxin toxicity reducer in the feed is 0.015 wt.% to 0.03 wt.%, and the basic feed can be pig feed or broiler feed, etc., depending on the specific type of livestock or poultry being targeted.

[0035] The aforementioned mycotoxin toxicity reducers can be added to the feed of livestock and poultry such as pigs or broilers as additives during the breeding process. They are used to regulate mitochondrial calcium transporters, alleviate mitochondrial calcium overload, and improve mitochondrial function, thereby mitigating the toxic effects of mycotoxins.

[0036] The three components in the above formulation were screened using techniques such as network pharmacology, high-throughput screening, and molecular docking. Currently, there are no reports on the combined use of peanut shell extract, rutin, and terpinene-4-ol to alleviate the toxicity of mycotoxins in feed. However, this invention has experimentally discovered that peanut shell extract, rutin, and terpinene-4-ol exhibit significant interactive effects in alleviating mitochondrial calcium overload and improving mitochondrial function, demonstrating a positive and significant synergistic effect. This can significantly alleviate the toxic effects of mycotoxins in feed on livestock and poultry, and improve their production performance and health.

[0037] The following examples demonstrate the specific application effects of the aforementioned mycotoxin toxicity reducer. The materials used in each of the following examples can be commercially available products or prepared using methods reported in existing technologies, and are not limited thereto.

[0038] Example 1

[0039] In this embodiment, the mycotoxin toxicity reducer is prepared according to the following steps 1 and 2:

[0040] Step 1: Place dried peanut shell powder in a flask, add 70% ethanol at a mass-to-volume ratio of 1:20, heat to about 60°C with stirring and extract for 1.6 h. After filtering to separate the filter cake and filtrate, the first extraction is completed. Add the same mass ratio of 70% ethanol to the filter cake and repeat the extraction twice. Combine the filtrates obtained from the three extractions and concentrate them. Add ethanol for alcohol precipitation, let stand overnight and filter. Evaporate the filtrate until there is no ethanol odor and then dry to obtain the peanut shell extract used in this example.

[0041] Step 2: Take 30 kg of peanut shell extract, 2 kg of rutin, and 1 kg of terpinene-4-ol, mix them thoroughly to obtain a mycotoxin toxicity attenuator (MTA). The addition amount in piglet feed is 0.03 wt.%. For ease of subsequent description, this experimental group is referred to as the MTA group.

[0042] One hundred and ninety-two Duroc-Landrace-Landrace-Bloodline piglets with an average weight of 12.8 kg were randomly divided into eight groups: ① Control group (basal diet contaminated with mycotoxins); ② Peanut shell extract group; ③ Rutin group; ④ Terpinene-4-ol group; ⑤ Rutin + Terpinene-4-ol group; ⑥ Peanut shell extract + Terpinene-4-ol group; ⑦ Peanut shell extract + Rutin group; ⑧ MTA group. The specific procedures for each group are as follows:

[0043] ① Control group: Feed contaminated with mycotoxins, the contents of aflatoxin B1, vomitoxin, zearalenone and fumonisin in the feed were 7, 852, 146 and 3960 μg / kg, respectively;

[0044] ② Peanut shell extract group: In addition to the control group, peanut shell extract was added alone to the same mycotoxin-contaminated feed;

[0045] ③ Rutin group: Rutin was added alone to the same mycotoxin-contaminated feed, based on the control group.

[0046] ④ Terpinene-4-ol group: Terpinene-4-ol was added alone to the same mycotoxin-contaminated feed, based on the control group.

[0047] ⑤ Rutin + Terpinene-4-ol group: Based on the control group, rutin and terpinene-4-ol were added separately to the same mycotoxin-contaminated feed.

[0048] ⑥ Peanut shell extract + terpinene-4-ol group: In addition to the control group, peanut shell extract and terpinene-4-ol were added separately to the same mycotoxin-contaminated feed.

[0049] ⑦ Peanut shell extract + rutin group: In addition to the control group, peanut shell extract and rutin were added separately to the same mycotoxin-contaminated feed.

[0050] ⑧MTA group: This group includes the aforementioned MTA composition from this embodiment, containing all three components, with an MTA addition amount of 0.03 wt.% in the feed.

[0051] Except for the control group and the MTA group, the content of each component added in groups ② to ⑦ above is consistent with the content of the corresponding component in the feed of the MTA group.

[0052] For the eight groups mentioned above, each group had three replicates, with eight piglets per replicate. The experimental period was 35 days, and the average daily weight gain and feed conversion ratio were calculated. After the feeding trial, intestinal, liver, and blood samples were collected from each group of pigs after slaughter for testing. The results of each indicator are as follows: Figure 1 and Figure 2 As shown.

[0053] Depend on Figure 1 As can be seen, compared with the control group, there were no significant differences in daily weight gain, feed conversion ratio, diarrhea rate, villus height / crypt depth (villus-crypt ratio), serum diamine oxidase (DAO), serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT), serum total antioxidant capacity (TAC) and malondialdehyde (MDA), and serum TNF-α in the single-drug groups ②-④ and the combination groups of any two components ⑤-⑦ (P>0.05). This indicates that single or two-component combinations cannot effectively alleviate mycotoxin-induced growth inhibition, intestinal and liver damage, oxidative stress, and inflammation. However, when the three components were used in combination (MTA group), all of the above indicators were significantly improved (P<0.05). Bliss independence model analysis showed that the three components had a very strong synergistic effect, which far exceeded the simple sum of the effects of the individual drugs. Peanut shell extract, rutin, and terpinene-4-ol, in a specific ratio, had a significant synergistic effect on alleviating mycotoxin-induced growth inhibition, intestinal damage, liver function damage, oxidative stress, and inflammation, producing unexpected technical effects.

[0054] Depend on Figure 2 As can be seen, compared with the control group, there were no significant differences in MCU protein expression, mitochondrial calcium content, mitochondrial membrane potential (ΔΨm), mitochondrial ROS, respiratory chain complex I activity, cellular oxygen consumption rate (OCR), basal respiration (BR), maximal respiration (MR), and ATP production in the single-drug groups ②-④ and the groups with any two components ⑤-⑦ (P>0.05). Furthermore, morphological changes of intestinal and liver mitochondrial damage (such as swelling, vacuolation, and cristae damage) were observed in groups ①-⑦. This indicates that neither a single component nor a combination of two components can effectively alleviate mycotoxin-induced mitochondrial functional damage. Compared with the control group, the MTA group showed significantly decreased MCU protein expression, mitochondrial calcium content, and mitochondrial ROS (P<0.05), and significantly increased mitochondrial membrane potential and respiratory chain complex activity (P<0.05); it also enhanced mitochondrial oxidative phosphorylation, manifested as a significant increase in cellular oxygen consumption rate (OCR), basal respiration (BR), maximal respiration (MR), and ATP production (P<0.05). Bliss independence model analysis showed that the three components had a very strong synergistic effect, far exceeding the simple sum of the effects of each individual drug. In summary, peanut shell extract, rutin, and terpinene-4-ol, in a specific ratio, exhibited a significant synergistic effect in alleviating mycotoxin-induced mitochondrial calcium overload and improving mitochondrial function, producing unexpected technical benefits.

[0055] Example 2

[0056] In this embodiment, the mycotoxin toxicity reducer is prepared according to the following steps 1 and 2:

[0057] Step 1: Crush dried peanut shells into small pieces using a mechanical method, pass them through a 60-mesh sieve, weigh the peanut shell powder and place it in a three-necked flask. Add 70% ethanol at a material-to-liquid mass ratio of 1:15, and add polyvinylpyrrolidone (PVP) at a peanut shell powder mass ratio of 0.025:1. Extract at 40°C for 7 hours. After vacuum filtration, the first extraction is complete, yielding a filtrate and a filter cake. The filter cake is then re-added with the same mass ratio of PPV and 70% ethanol as in the first extraction, and the extraction is repeated twice. The filtrates from the three extractions are combined. The filtrate is rotary evaporated to a paste state. The paste is crushed, washed once with water, and dried to obtain the peanut shell extract used in this example.

[0058] Step 2: Take 35 kg of peanut shell extract, 3 kg of rutin, and 2 kg of terpinene-4-ol, mix them thoroughly to obtain a mycotoxin toxicity attenuator (MTA). The addition amount in pig feed is 0.02 wt.%. For ease of subsequent description, this experimental group is referred to as the MTA group.

[0059] Ninety-six Jinhua pigs with an average weight of 52 kg were randomly divided into eight groups: ① Control group (basal diet contaminated with mycotoxins); ② Peanut shell extract group; ③ Rutin group; ④ Terpinene-4-ol group; ⑤ Rutin + Terpinene-4-ol group; ⑥ Peanut shell extract + Terpinene-4-ol group; ⑦ Peanut shell extract + Rutin group; ⑧ MTA group. The specific procedures for each group are as follows:

[0060] ① Control group: Feed contaminated with mycotoxins, the contents of aflatoxin B1, vomitoxin, zearalenone and fumonisin in the feed were 12, 826, 184 and 3237 μg / kg, respectively;

[0061] ② Peanut shell extract group: In addition to the control group, peanut shell extract was added alone to the same mycotoxin-contaminated feed;

[0062] ③ Rutin group: Rutin was added alone to the same mycotoxin-contaminated feed, based on the control group.

[0063] ④ Terpinene-4-ol group: Terpinene-4-ol was added alone to the same mycotoxin-contaminated feed, based on the control group.

[0064] ⑤ Rutin + Terpinene-4-ol group: Based on the control group, rutin and terpinene-4-ol were added separately to the same mycotoxin-contaminated feed.

[0065] ⑥ Peanut shell extract + terpinene-4-ol group: In addition to the control group, peanut shell extract and terpinene-4-ol were added separately to the same mycotoxin-contaminated feed.

[0066] ⑦ Peanut shell extract + rutin group: In addition to the control group, peanut shell extract and rutin were added separately to the same mycotoxin-contaminated feed.

[0067] ⑧MTA group: This group includes the aforementioned MTA composition from this embodiment, containing all three components, with an MTA addition amount of 0.02 wt.% in the feed.

[0068] Except for the control group and the MTA group, the content of each component added in groups ② to ⑦ above is consistent with the content of the corresponding component in the feed of the MTA group.

[0069] For the eight groups mentioned above, each group had three replicates, with four Jinhua pigs per replicate. The experimental period was 56 days, and the average daily weight gain and feed conversion ratio were calculated. After the feeding trial, intestinal, liver, and blood samples were collected from each group of pigs after slaughter for testing. The results of each indicator are as follows: Figure 3 and Figure 4 As shown.

[0070] Depend on Figure 3 As can be seen, compared with the control group, there were no significant differences in daily weight gain, feed conversion ratio, chorionic villus ratio, serum diamine oxidase (DAO), serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT), serum total antioxidant capacity (TAC) and malondialdehyde (MDA), and serum TNF-α in the single-drug groups ②-④ and the combination groups of any two components ⑤-⑦ (P>0.05). This indicates that single or two-component combinations cannot effectively alleviate mycotoxin-induced growth inhibition, intestinal and liver damage, oxidative stress, and inflammation. However, when the three components were used in combination (MTA group), all of the above indicators were significantly improved (P<0.05). Bliss independence model analysis showed that the three components had a very strong synergistic effect, far exceeding the simple sum of the effects of the individual drugs. Peanut shell extract, rutin, and terpinene-4-ol, in a specific ratio, showed a significant synergistic effect in alleviating mycotoxin-induced growth inhibition, intestinal damage, liver function damage, oxidative stress, and inflammation, producing unexpected technical effects.

[0071] Depend on Figure 4As can be seen, compared with the control group, there were no significant differences in MCU expression, mitochondrial calcium content, mitochondrial membrane potential (ΔΨm), mitochondrial ROS, respiratory chain complex I activity, cellular oxygen consumption rate (OCR), basal respiration (BR), maximal respiration (MR), and ATP production in the single-drug groups ②-④ and the groups with any two components ⑤-⑦ (P>0.05); and morphological changes of intestinal and liver mitochondrial damage (such as swelling, vacuolation, and cristae damage) were observed in groups ①-⑦. This indicates that neither a single component nor a combination of two components can effectively alleviate mycotoxin-induced mitochondrial functional damage. Compared with the control group, the MTA group showed significantly decreased MCU protein expression, mitochondrial calcium content, and mitochondrial ROS (P<0.05), and significantly increased mitochondrial membrane potential and respiratory chain complex activity (P<0.05); it also enhanced mitochondrial oxidative phosphorylation, as evidenced by significantly increased cellular oxygen consumption rate (OCR), basal respiration (BR), maximal respiration (MR), and ATP production (P<0.05). Bliss independence model analysis showed that the three components had a very strong synergistic effect, far exceeding the simple sum of the effects of each individual drug. In summary, peanut shell extract, rutin, and terpinene-4-ol, in a specific ratio, exhibited a significant synergistic effect in alleviating mycotoxin-induced mitochondrial calcium overload and improving mitochondrial function, producing unexpected technical benefits.

[0072] Example 3

[0073] In this embodiment, the mycotoxin toxicity reducer is prepared according to the following steps 1 and 2:

[0074] Step 1: Crush dried peanut shells into small pieces using a mechanical method, pass them through a 60-mesh sieve, weigh the peanut shell powder and place it in a three-necked flask. Add 70% ethanol at a material-to-liquid mass ratio of 1:15, and add polyethylene glycol 200 at a peanut shell powder mass ratio of 0.02:1. Extract at 40℃ for 7 hours. After vacuum filtration, the first extraction is complete, yielding filtrate and filter cake. The filter cake is then re-added with the same mass ratio of polyethylene glycol 200 and 70% ethanol as in the first extraction, and extracted three times. Combine the filtrates from the three extractions. Rotary evaporate the filtrate to a paste state. Crush the paste, wash it once with water, and dry it to obtain the peanut shell extract used in this example.

[0075] Step 2: Take 40 kg of peanut shell extract, 5 kg of rutin, and 3 kg of terpinene-4-ol, mix them thoroughly to obtain a mycotoxin toxicity attenuator (MTA). The addition amount in broiler feed is 0.015 wt.%. For ease of subsequent description, this experimental group is referred to as the MTA group.

[0076] Three hundred and sixty one-day-old yellow-feathered broilers were randomly divided into eight groups: ① Control group (basal diet contaminated with mycotoxins); ② Peanut shell extract group; ③ Rutin group; ④ Terpinene-4-ol group; ⑤ Rutin + Terpinene-4-ol group; ⑥ Peanut shell extract + Terpinene-4-ol group; ⑦ Peanut shell extract + Rutin group; ⑧ MTA group. The specific procedures for each group are as follows:

[0077] ① Control group: Feed contaminated with mycotoxins, the contents of aflatoxin B1, vomitoxin, zearalenone and T-2 toxin in the feed were 9, 2369, 465 and 418 μg / kg, respectively;

[0078] ② Peanut shell extract group: In addition to the control group, peanut shell extract was added alone to the same mycotoxin-contaminated feed;

[0079] ③ Rutin group: Rutin was added alone to the same mycotoxin-contaminated feed, based on the control group.

[0080] ④ Terpinene-4-ol group: Terpinene-4-ol was added alone to the same mycotoxin-contaminated feed, based on the control group.

[0081] ⑤ Rutin + Terpinene-4-ol group: Based on the control group, rutin and terpinene-4-ol were added separately to the same mycotoxin-contaminated feed.

[0082] ⑥ Peanut shell extract + terpinene-4-ol group: In addition to the control group, peanut shell extract and terpinene-4-ol were added separately to the same mycotoxin-contaminated feed.

[0083] ⑦ Peanut shell extract + rutin group: In addition to the control group, peanut shell extract and rutin were added separately to the same mycotoxin-contaminated feed.

[0084] ⑧ MTA group: This group includes the aforementioned composition MTA from this embodiment, containing all three components, with an MTA addition amount of 0.015 wt.% in the feed.

[0085] Except for the control group and the MTA group, the content of each component added in groups ② to ⑦ above is consistent with the content of the corresponding component in the feed of the MTA group.

[0086] For the eight groups mentioned above, each group had three replicates, with 15 birds per replicate, and the experimental period was 70 days. After the feeding trial, intestinal, liver, and blood samples were collected from each group of broilers after slaughter for testing. The results of each indicator are as follows: Figure 5 and Figure 6 As shown.

[0087] Depend on Figure 5 It is evident that, compared with the control group, there were no significant differences in daily weight gain, feed conversion ratio, chorionic villus ratio, serum diamine oxidase (DAO), serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT), serum total antioxidant capacity (TAC) and malondialdehyde (MDA), and serum TNF-α in the single-drug groups ②-④ and any two-component combination groups ⑤-⑦ (P>0.05). This indicates that single or two-component combinations cannot effectively alleviate mycotoxin-induced growth inhibition, intestinal and liver damage, oxidative stress, and inflammation. However, when all three components were used in combination (MTA group), all of the above indicators were significantly improved (P<0.05). Bliss independence model analysis showed that the three components had a very strong synergistic effect, far exceeding the simple sum of the effects of the individual drugs. Peanut shell extract, rutin, and terpinene-4-ol, in specific ratios, showed significant synergistic effects in alleviating mycotoxin-induced growth inhibition, intestinal damage, liver function damage, oxidative stress, and inflammation, producing unexpected technical effects.

[0088] Depend on Figure 6 As can be seen, compared with the control group, there were no significant differences in MCU expression, mitochondrial calcium content, mitochondrial membrane potential (ΔΨm), mitochondrial ROS, respiratory chain complex I activity, cellular oxygen consumption rate (OCR), basal respiration (BR), maximal respiration (MR), and ATP production in the single-drug groups ②-④ and the groups with any two components ⑤-⑦ (P>0.05); and morphological changes of intestinal and liver mitochondrial damage (such as swelling, vacuolation, and cristae damage) were observed in groups ①-⑦. This indicates that neither a single component nor a combination of two components can effectively alleviate mycotoxin-induced mitochondrial functional damage. Compared with the control group, the MTA group showed significantly decreased MCU protein expression, mitochondrial calcium content, and mitochondrial ROS (P<0.05), and significantly increased mitochondrial membrane potential and respiratory chain complex activity (P<0.05); it also enhanced mitochondrial oxidative phosphorylation, as evidenced by significantly increased cellular oxygen consumption rate (OCR), basal respiration (BR), maximal respiration (MR), and ATP production (P<0.05). Bliss independence model analysis showed that the three components had a very strong synergistic effect, far exceeding the simple sum of the effects of each individual drug. In summary, peanut shell extract, rutin, and terpinene-4-ol, in a specific ratio, exhibited a significant synergistic effect in alleviating mycotoxin-induced mitochondrial calcium overload and improving mitochondrial function, producing unexpected technical benefits.

[0089] The embodiments described above are merely some preferred implementations of the present invention and are not intended to limit the invention. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention. Therefore, all technical solutions obtained through equivalent substitution or transformation fall within the protection scope of the present invention.

Claims

1. A mycotoxin toxicity reducer, characterized in that, The mycotoxin toxicity reducer, by weight, is composed of 30-40 parts peanut shell extract, 2-5 parts rutin, and 1-3 parts terpinene-4-ol; The peanut shell extract was obtained from peanut shells by ethanol extraction. The extraction method is as follows: dried peanut shells are crushed and extracted multiple times at a temperature of 40~60℃ using ethanol as a solvent. The extracts are combined, concentrated, purified and dried to obtain the peanut shell extract.

2. The mycotoxin toxicity reducer according to claim 1, characterized in that, During the extraction process using ethanol as a solvent, an extraction aid is required, including polyvinylpyrrolidone or polyethylene glycol 200; the purification operation includes alcohol precipitation or water washing.

3. A feed containing a mycotoxin toxicity reducer as described in claim 1 or 2.

4. The feed as described in claim 3, characterized in that, The amount of mycotoxin toxicity reducer added to the feed is 0.015 wt.% to 0.03 wt.%.

5. The feed as described in claim 3, characterized in that, The feed is pig feed or broiler feed.

6. The application of a mycotoxin toxicity reducer as described in claim 1 or 2 in the preparation of livestock and poultry feed, characterized in that, It is used to alleviate the toxic effects of mycotoxins by regulating mitochondrial calcium transporters, relieving mitochondrial calcium overload, and improving mitochondrial function.

7. The application as described in claim 6, characterized in that, The mycotoxin toxicity reducer is added as an additive to the feed for livestock and poultry.

8. The application as described in claim 6, characterized in that, The livestock and poultry mentioned are pigs or broiler chickens.

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