Zanthoxylum bungeanum leaf-vinasse fermentation composition

Abstract

The invention discloses a Zanthoxylum bungeanum leaf-vinasse fermentation composition which comprises the following raw materials: vinasse, Zanthoxylum bungeanum leaves and a mixed microbial agent, the mixed bacterial agent is lactobacillus brevis, lactobacillus plantarum, bacillus subtilis and saccharomyces cerevisiae; the zanthoxylum bungeanum leaf-vinasse fermentation composition is obtained by anaerobic solid-state fermentation of the raw materials, that is, vinasse and zanthoxylum bungeanum leaves are crushed and uniformly mixed, then the mixture is uniformly mixed with a mixed bacterial agent, and sealed anaerobic fermentation is performed for 4-6 days at the fermentation temperature of 35-40 DEG C, so that the zanthoxylum bungeanum leaf-vinasse fermentation composition is obtained. By mixing the bunge pricklyash leaves with the vinasse and then inoculating the mixed microbial inoculum for fermentation, the antioxidant activity of the bunge pricklyash leaves is more reserved, the nutritional value is higher, the method has positive significance for replacing antibiotics and protecting animal health, the weight loss ratio is reduced after the mixed microbial inoculum is added, the content of soluble sugar is increased, and true protein is not obviously lost; higher nutritional value can be provided for animals.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a Zanthoxylum bungeanum leaf-vinasse fermentation composition. Background Art

[0002] The fruit of Zanthoxylum bungeanum maxim is small in size. In the process of collecting raw materials of Zanthoxylum bungeanum, the fruit of Zanthoxylum bungeanum is often picked together with the branches and leaves of Zanthoxylum bungeanum. The raw materials for making Zanthoxylum bungeanum in the food industry are mainly the peel of Zanthoxylum bungeanum. In the process of producing Zanthoxylum bungeanum, a large number of by-products such as Zanthoxylum bungeanum maxim leaves are produced. These by-products are often discarded or incinerated, which eventually causes environmental pollution and waste of resources. Zanthoxylum bungeanum maxim leaves contain rich nutrients such as protein, carbohydrates, phenolic compounds, vitamins, etc. Directly discarding them is a major waste of resources. Therefore, some studies have explored the inclusion of Zanthoxylum bungeanum maxim leaves in animal feed, thereby recycling resources and solving the problem of feed resource shortage. However, the high anti-nutritional factors of Zanthoxylum bungeanum maxim leaves will have adverse effects on animals, which greatly limits the promotion and application of production by-products such as Zanthoxylum bungeanum maxim leaves.

[0003] Distillers' grains (DG) are the residue left after alcohol is distilled from grains. These grains are rich in nutrients, but their direct processing causes environmental pollution and waste of resources. Given the high protein content in DG, the current popular treatment method is to use it as an additive in animal feed. However, the high acid content in DG may easily induce acidosis in animals fed directly. In addition, wet DG can deteriorate rapidly in the air, producing excessive mycotoxins, which affect the health of livestock. Therefore, DG should be treated using appropriate methods before being used as feed to improve its safety and nutritional value.

[0004] At present, traditional feed resources are insufficient to meet the needs of animal feeding, and new unconventional feed resources such as pepper leaves and distiller's grains need to be developed. However, various problems with these resources still exist, hindering their widespread promotion and utilization. Summary of the invention

[0005] An object of the present invention is to solve at least the above problems and / or disadvantages and to provide at least the advantages which will be described hereinafter.

[0006] In order to achieve these purposes and other advantages of the present invention, a Zanthoxylum leaf-vintner's lees fermentation composition is provided, which comprises the following raw materials: vintner's lees, Zanthoxylum leaf and mixed bacterial agent;

[0007] The mixed bacterial agent comprises Levilactobacillus brevis, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae.

[0008] Preferably, the dry mass ratio of the wine lees to the Zanthoxylum bungeanum leaves is 1:0.5-2; and the dosage of the mixed bacterial agent is 5-15wt% of the total mass of the wine lees and the Zanthoxylum bungeanum leaves.

[0009] Preferably, the Zanthoxylum bungeanum leaf-vinasse fermentation composition is obtained by anaerobic solid-state fermentation of the raw materials.

[0010] Preferably, in the mixed bacterial agent, Lactobacillus breve, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae are composed in a mass ratio of 1-3:1-3:1-3:1-3.

[0011] Preferably, the mixed bacterial agent is Lactobacillus breve, Bacillus subtilis and Saccharomyces cerevisiae.

[0012] Preferably, in the mixed bacterial agent, Lactobacillus breve, Bacillus subtilis and Saccharomyces cerevisiae are composed in a mass ratio of 1-3:1-3:1-3.

[0013] Preferably, the mixed bacterial agent is Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae.

[0014] Preferably, in the mixed bacterial agent, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae are composed in a mass ratio of 1-3:1-3:1-3.

[0015] Preferably, the specific steps of the anaerobic solid-state fermentation are: crushing the vinasse and Zanthoxylum bungeanum leaves into 20-50 meshes and mixing them evenly, then mixing them with the mixed bacterial agent, stirring them evenly, adjusting the water content of the raw materials to 50-70wt%, and sealing the anaerobic fermentation for 4-6 days at a fermentation temperature of 35-40°C to obtain a Zanthoxylum bungeanum-vinasse fermentation composition.

[0016] Preferably, the raw materials also include additives; the additives are obtained by mixing stachyose, trehalose, magnesium sulfate, manganese sulfate and diammonium hydrogen citrate in a mass ratio of 1-3:0.5-1.5:0.1-0.3:0.1-0.3:0.3-0.8; and the amount of the additive added is 8wt% of the total mass of the lees and the prickly ash leaves.

[0017] Preferably, the specific steps of the anaerobic solid-state fermentation are: mixing stachyose, trehalose, magnesium sulfate, manganese sulfate and diammonium hydrogen citrate to obtain an additive, adding 5 to 10 times the mass of distilled water, stirring evenly, and obtaining an additive solution; crushing vinasse and Zanthoxylum bungeanum leaves to 20 to 50 meshes and mixing evenly, then adding the additive solution, stirring evenly, irradiating the obtained mixed material, and then mixing with the mixed bacterial agent, stirring evenly, adjusting the water content of the raw material to 50 to 70wt%, and sealing anaerobic fermentation for 4 to 6 days at a fermentation temperature of 35 to 40°C to obtain a Zanthoxylum bungeanum-vinasse fermentation composition.

[0018] Preferably, the irradiation uses 60 cobalt gamma rays, the irradiation dose is 10 to 30 Gy, and the dose rate is 5 to 15 Gy / min.

[0019] A use of the Zanthoxylum bungeanum leaf-vinasse fermentation composition as described above in an animal feed additive.

[0020] The present invention at least includes the following beneficial effects: the present invention mixes Zanthoxylum leaves with distiller's grains, and then inoculates a mixed bacterial agent for fermentation, so that the antioxidant activity of Zanthoxylum leaves is more retained and the nutritional value is higher. After Zanthoxylum leaves are mixed with distiller's grains, their nutritional structure is more reasonable and suitable for the growth and fermentation of microorganisms, but not inoculating a mixed bacterial agent will lead to the growth of miscellaneous bacteria, increase the weight loss rate, destroy the nutritional structure, and lose more antioxidant properties. The present invention adopts Bacillus subtilis, Saccharomyces cerevisiae and Lactobacillus breve and / or Lactobacillus plantarum to prepare a mixed bacterial agent. By inoculating three mixed bacterial agents, the microbial community structure in the solid-state fermentation system is changed, the growth of harmful microorganisms is effectively controlled, the content of anti-nutritional factors is reduced, the antioxidant activity is improved, DM loss is reduced, the soluble sugar content is increased, and there is no obvious loss of true protein, which can provide animals with higher nutritional value. The mixed bacterial agent inoculated with breve lactobacillus, plantarum lactobacillus, bacillus subtilis and saccharomyces cerevisiae exhibits the most comprehensive and optimal solid-state fermentation performance, and its antioxidant activity can be more retained, which has positive significance for replacing antibiotics and protecting animal health, and is of great significance for the comprehensive recycling of Zanthoxylum bungeanum leaves and distiller's grains in the production of unconventional feed resources.

[0021] Other advantages, objectives and features of the present invention will be embodied in part through the following description, and in part will be understood by those skilled in the art through study and practice of the present invention. DETAILED DESCRIPTION

[0022] The present invention is described in further detail below so that those skilled in the art can implement it according to the description.

[0023] It should be understood that the terms such as “having”, “including” and “comprising” used herein do not exclude the existence or addition of one or more other elements or combinations thereof.

[0024] In the following examples, the raw materials and test materials used, unless otherwise specified, were purchased from commercial channels; the experimental methods used, unless otherwise specified, were conventional methods.

[0025] In the following examples, Zanthoxylum bungeanum leaves were from a company in Mianyang, China, and distiller's grains were from a winery in Mianyang, China.

[0026] In the following examples, the saccharomyces cerevisiae used was CICC 1202, purchased from the China Industrial Microbiological Culture Collection Administration Center; Bacillus subtilis was CCTCC NO: M2024804 (Chinese patent CN2024107543110); Levilactobacillus brevis was CICC20297, purchased from the China Industrial Microbiological Culture Collection Administration Center; Lactobacillus plantarum was CICC 24936, purchased from China Industrial Microbiological Culture Collection Center; pre-screening was carried out in YPD medium (Saccharomyces cerevisiae), LB medium (Bacillus subtilis), and MRS medium (Lactobacillus breve and Lactobacillus plantarum), respectively, with the initial pH of the medium being 6.5, 7.0, and 5.7, respectively. Saccharomyces cerevisiae and Bacillus subtilis were cultured at 37°C, 110 rpm under shaking conditions for 24 h, while Lactobacillus plantarum and Lactobacillus breve were cultured at 37°C without shaking for 24 h.

[0027] Example 1

[0028] A Zanthoxylum leaf-vinasse fermentation composition comprises the following raw materials: vinasse, Zanthoxylum leaf, and mixed bacterial agent; wherein, based on dry weight, the mass ratio of vinasse to Zanthoxylum leaf is 1:1, and the amount of the mixed bacterial agent is 10wt% of the total mass of vinasse and Zanthoxylum leaf;

[0029] The mixed bacterial agent is composed of Lactobacillus breve, Bacillus subtilis and Saccharomyces cerevisiae in a mass ratio of 1:1:1;

[0030] The preparation method of the Zanthoxylum leaf-vintner's lees fermentation composition is as follows: crushing the vintner's lees and the Zanthoxylum leaf into 30 meshes and then mixing them evenly, then mixing them with the mixed bacterial agent, stirring evenly, adjusting the water content of the raw materials to 60wt%, putting them into a fermentation bag, setting the fermentation temperature to 37°C±0.5°C, and conducting sealed anaerobic fermentation for 5 days to obtain the Zanthoxylum leaf-vintner's lees fermentation composition.

[0031] Example 2

[0032] This embodiment is substantially the same as embodiment 1, except that the mixed bacterial agent of this embodiment is composed of Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae in a mass ratio of 1:1:1.

[0033] Example 3

[0034] This embodiment is basically the same as Embodiment 1, except that the mixed bacterial agent of this embodiment is composed of Lactobacillus breve, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae in a mass ratio of 1:1:1:1.

[0035] Example 4

[0036] A Zanthoxylum leaf-vintner's grain fermentation composition comprises the following raw materials: vintner's grains, Zanthoxylum leafs, mixed bacterial agent and additives; wherein, based on dry weight, the mass ratio of vintner's grains to Zanthoxylum leafs is 1:1, the amount of the mixed bacterial agent is 10wt% of the total mass of the vintner's grains and Zanthoxylum leafs, and the amount of the additive is 8wt% of the total mass of the vintner's grains and Zanthoxylum leafs;

[0037] The mixed bacterial agent is composed of Lactobacillus breve, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae in a mass ratio of 1:1:1:1; the additive is obtained by mixing stachyose, trehalose, magnesium sulfate, manganese sulfate and diammonium hydrogen citrate in a mass ratio of 2:1:0.2:0.2:0.5;

[0038] The preparation method of the Zanthoxylum bungeanum-vinasse fermentation composition comprises the following steps: mixing stachyose, trehalose, magnesium sulfate, manganese sulfate and diammonium hydrogen citrate in a mass ratio of 2:1:0.2:0.2:0.5, adding the mixture to 8 times the mass of distilled water, stirring evenly, and obtaining an additive solution; crushing the vinasse and Zanthoxylum bungeanum to 30 mesh and mixing evenly, adding the additive solution, stirring evenly, irradiating the obtained mixture, mixing the mixture with a mixed bacterial agent, stirring evenly, adjusting the water content of the raw material to 60wt%, loading the raw material into a fermentation bag, setting the fermentation temperature to 37°C±0.5°C, and conducting sealed anaerobic fermentation for 5 days, and obtaining the Zanthoxylum bungeanum-vinasse fermentation composition; wherein the irradiation adopts 60 cobalt gamma rays, the irradiation dose is 20Gy, and the dose rate is 10Gy / min.

[0039] Example 5

[0040] This embodiment is basically the same as Example 4, except that this embodiment does not perform irradiation treatment, that is, the preparation method of the Zanthoxylum leaf-vintner's lees fermentation composition of this embodiment is: stachyose, trehalose, magnesium sulfate, manganese sulfate and diammonium hydrogen citrate are mixed in a mass ratio of 2:1:0.2:0.2:0.5, added to 8 times the mass of distilled water, and stirred evenly to obtain an additive solution; the vintner's lees and Zanthoxylum leaf are crushed to 30 mesh and then mixed evenly, and then the additive solution is added, stirred evenly, and then mixed with the mixed bacterial agent, stirred evenly, the water content of the raw material is adjusted to 60wt%, and the raw material is loaded into a fermentation bag. The fermentation temperature is 37°C±0.5°C, and the sealed anaerobic fermentation is carried out for 5 days to obtain the Zanthoxylum leaf-vintner's lees fermentation composition.

[0041] Comparative Example 1

[0042] This comparative example is basically the same as Example 1, except that the raw materials of this comparative example do not contain distiller's grains, but only have Zanthoxylum bungeanum leaves and mixed bacterial agents to obtain a Zanthoxylum bungeanum leaf fermentation composition.

[0043] Comparative Example 2

[0044] This comparative example is basically the same as Example 2, except that the raw materials of this comparative example do not contain distiller's grains, but only have Zanthoxylum bungeanum leaves and mixed bacterial agents to obtain a Zanthoxylum bungeanum leaf fermentation composition.

[0045] Comparative Example 3

[0046] This comparative example is basically the same as Example 3, except that the raw materials of this comparative example do not contain distiller's grains, but only have Zanthoxylum bungeanum leaves and mixed bacterial agents to obtain a Zanthoxylum bungeanum leaf fermentation composition.

[0047] Comparative Example 4

[0048] This comparative example is basically the same as Example 1, except that the raw materials of this comparative example do not contain Zanthoxylum bungeanum leaves, but only contain distiller's grains and mixed bacterial agents to obtain a distiller's grains fermentation composition.

[0049] Comparative Example 5

[0050] This comparative example is basically the same as Example 2, except that the raw materials of this comparative example do not contain Zanthoxylum bungeanum leaves, but only contain distiller's grains and mixed bacterial agents to obtain a distiller's grains fermentation composition.

[0051] Comparative Example 6

[0052] This comparative example is basically the same as Example 3, except that the raw materials of this comparative example do not contain Zanthoxylum bungeanum leaves, but only contain distiller's grains and mixed bacterial agents to obtain a distiller's grains fermentation composition.

[0053] The various indicators of the fermented compositions obtained in the examples and comparative examples were examined, including: ① conventional indicators (dry matter content DM, weight loss rate DML, dry weight retention rate DMR, neutral detergent fiber content NDF, water-soluble carbohydrate content WSC, pH); ② anti-nutritional factors (α-sanshool content, α-sanshool degradation rate, tannin content, tannin degradation rate, phytic acid content, phytic acid degradation rate); ③ protein content (true protein content TPN, ammonia nitrogen NH 3 -N); ④ Functional nutrients (antioxidant activity, organic acid content).

[0054] The changes in the physicochemical properties and the contents of various substances of the compositions of Examples 1 to 3 and Comparative Examples 1 to 6 before and after fermentation are shown in Table 1, the changes in the organic acid content are shown in Table 2, the changes in the anti-nutritional factor content are shown in Table 3, and the changes in the antioxidant activity are shown in Table 4. In the table, the CK group is a group not inoculated with a mixed bacterial agent; RM is a raw material (before fermentation); experimental groups: Group A is inoculated with Lactobacillus breve, Bacillus subtilis and Saccharomyces cerevisiae, Group B is inoculated with Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae, and Group C is inoculated with Lactobacillus breve, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae; DPPH: scavenging activity of DPPH free radicals; ABTS: ABTS + Free radical scavenging activity; “-” means no or not detected.

[0055] (1) Changes in the fermentation of Zanthoxylum bungeanum leaves when inoculated with different combinations of bacterial agents.

[0056] After natural fermentation without inoculation of mixed bacteria (CK group), pH and NH 3 -N (Ammoniacal Nitrogen) was higher than that of the raw material, while WSC (Water Soluble Carbohydrates), tannins, phytic acid, α-sanshool and antioxidant activity were all lower than that of the raw material.

[0057] Compared with the RM group, the pH, tannin, phytic acid, α-sanshool and antioxidant activity were all reduced in the experimental groups (the DPPH scavenging activity of group C was the most retained, and the ABTS scavenging activity of group B was the most retained), NH 3 -N increased, TPN (True protein content) and NDF content increased in both the experimental and CK groups.

[0058] Compared with the CK group, after inoculation with the mixed bacterial agent, the NH 3-N was higher, the antioxidant activity of the experimental groups was mostly higher than that of CK (the ABTS free radical scavenging ability of group B was 57.7% higher than that of group CK, and the ABTS of group C was lower than that of group CK), there was no significant difference in tannin content, the phytic acid content of groups B and C was significantly lower than that of CK (reduced by 67.6% and 62.3% relative to the raw material, and reduced by 51.2% and 41.9% relative to the CK content), the content of α-sanshool in the experimental groups was higher than that in the CK group, and at the same time, because the mixed bacterial agent utilized more carbon sources, the WSC content was further reduced.

[0059] (2) Changes in the inoculation of different bacterial agents when Zanthoxylum bungeanum leaves were mixed with distiller's grains for fermentation.

[0060] Compared with the raw material, the pH of the CK group increased, the true protein content increased, and the NH 3 -N did not change significantly, NDF increased, and the other parameters decreased.

[0061] Compared with the raw material, the experimental group 3 -N and ABTS scavenging abilities were increased, tannin, phytic acid and α-sanshool contents were decreased, WSC of group C was increased, WSC of groups A and B was slightly decreased, and TPN content had no significant change.

[0062] Compared with the CK group, the experimental group had lower pH, lower TPN content, and lower NH 3 -N, WSC, tannin, phytic acid, α-sanshool contents and antioxidant activity were higher, and DMR increased.

[0063] (3) The mixed fermentation of Zanthoxylum bungeanum leaves and distiller's grains was compared with the fermentation of Zanthoxylum bungeanum leaves alone.

[0064] The mixed fermentation experimental group had lower pH, lower TPN content, lower weight loss rate, and NH 3 -N and WSC were higher, the phytic acid degradation rate and tannin degradation rate were lower, the degradation rate of α-sanshool was partially improved (the mixed fermentation group B was higher than the single fermentation group, and the A and C groups were lower than the single fermentation group), the retention rate of DPPH scavenging ability was higher, and the ABTS scavenging ability changed from a decrease in single fermentation to an increase.

[0065] When Zanthoxylum leaves were mixed with distiller's grains for fermentation, the growth of microorganisms could be promoted without inoculation of bacterial agents, and the weight loss rate increased due to microbial activity. However, the weight loss rate was significantly reduced after inoculation of mixed bacterial agents, indicating that the inoculated mixed bacterial agents could inhibit the growth of miscellaneous bacteria and prevent excessive loss of nutrients. After inoculation of mixed bacterial agents, the WSC content increased, which may also be due to the inhibition of the growth of miscellaneous bacteria. At the same time, the growth metabolism of the inoculated mixed bacterial community decomposed some insoluble carbohydrates into soluble sugars.

[0066] The TPN content of the experimental group was lower than that of the CK group. The possible reason was that the weight loss rate of the CK group was higher, and many non-protein components were decomposed and utilized (such as the significantly reduced WSC content), resulting in a lower total weight, so the TPN content was relatively higher. Overall, the inoculation of mixed bacterial agents did not significantly increase or decrease TPN. 3 The increase in -N content may be caused by the degradation of other non-protein nitrogen.

[0067] After fermentation, the NDF content increased relative to the raw materials. The possible reason is that the degradation of other raw materials led to a decrease in total weight, while the NDF content changed relatively little, resulting in an increase in the proportion of total weight. Among them, the NDF of group A, which consisted of Zanthoxylum leaves, distiller's grains, and Zanthoxylum leaves + distiller's grains, was the lowest after fermentation. It is speculated that the microorganisms in group A have better NDF degradation capabilities.

[0068] As shown in Table 2, the raw material of Zanthoxylum bungeanum contains only a small amount of acetic acid and no other organic acids. After natural fermentation, the organic acid content did not change significantly, and only the acetic acid content increased. After inoculation with mixed bacterial agents, the contents of lactic acid and acetic acid increased significantly, especially the lactic acid content of Group B and Group C reached more than 40 mg / g, and the ratio of lactic acid to acetic acid was 2.37 and 2.18, respectively, which is close to the general silage requirements. Due to the production of liquor, microorganisms produce various organic acids, and there are organic acid residues in the lees. The lees raw materials contain various organic acids. After fermentation, the content of these organic acids further increases, especially the lactic acid content, which increases to 78.42 mg / g in Group B. After mixed fermentation of Zanthoxylum bungeanum and lees, the content of acetic acid and lactic acid in Group CK increased compared with the raw material, but the increase in acetic acid was much higher than that of lactic acid, resulting in a significant decrease in the ratio of lactic acid to acetic acid. In the experimental group, the acetic acid content was lower than 8 mg / g, the lactic acid content was higher than 35 mg / g, and the ratio of lactic acid to acetic acid was higher than 5.5, indicating that the inoculated mixed bacterial agent inhibited the production of acetic acid and promoted the production of lactic acid in the mixed raw materials. In addition, a significant increase in propionic acid content was observed in the CK group of Zanthoxylum leaves + lees, which may be caused by the growth of Clostridium.

[0069] Phytic acid and tannins are anti-nutritional compounds that chelate proteins and ions. Phytic acid in plant feeds can cause reproductive problems. Polygastric animals can degrade phytic acid through their intestinal microbiota, but monogastric animals (poultry, pigs, fish, humans) cannot. Phytic acid accumulation occurs by reducing protein and ions (Fe 3+ , Ca 2+ 、Zn 2+ Mg 2+) and thus harm animal health by reducing the bioavailability of tannins. Tannins are defensive metabolites evolved by plants to prevent infection by animals and pathogens and inhibit the growth and biodegradation of microorganisms. Tannins can bind to proteins to form complexes. Monogastric animals have poor tolerance to the astringent effects caused by tannin-protein complexes. α-Sanshool is a special anti-nutritional factor in Zanthoxylum bungeanum leaves. α-Sanshool has a long-chain unsaturated fat structure and is the main component that causes the tingling and numbness in Zanthoxylum bungeanum leaves. α-Sanshool also exhibits other physiological functions such as analgesia, anesthesia, antioxidant activity, and antibacterial and anti-inflammatory properties.

[0070] As shown in Table 3, the phytic acid degradation rates of groups B and C fermented with Zanthoxylum leaves alone were significantly improved. This may be related to the good phytic acid degradation ability of Lactobacillus plantarum. In addition, after inoculating mixed bacterial agents with Zanthoxylum leaves and distiller's grains as single substrates or mixed substrates, the degradation rates of tannins, phytic acid and α-sanshool in most experimental groups were lower than those in group CK. This may be because the inoculated microorganisms inhibited the growth of other microorganisms, while the uninoculated raw materials contained some microorganisms that could degrade tannins, phytic acid and α-sanshool, and their growth was not restricted, which could better degrade these substances. It is worth noting that when Zanthoxylum leaves were fermented alone and mixed, the degradation rates of α-sanshool in group B reached 48.81% and 53.93%, respectively, which were higher than those in other experimental groups, but only slightly lower than those in group CK fermented with Zanthoxylum leaves alone. This shows that the microorganisms in group B have good α-sanshool degradation ability. Although various anti-nutritional factors in Zanthoxylum leaves can be effectively degraded without inoculation of exogenous microorganisms, the excessive growth of harmful microorganisms (such as Escherichia coli) is harmful to solid-state fermentation.

[0071] After fermentation with distiller's grains as the sole raw material, except for the significant decrease in WSC content, the changes in other parameters in Table 1 were not significant. This shows that distiller's grains are difficult to ferment as a single substrate due to their high acidity and rich antibacterial substances, which hinders the rapid growth of most microorganisms. Its DPPH scavenging ability decreased, and there was no significant difference between the experimental group and the CK group, while the ABTS scavenging ability increased, which means that some antioxidant substances may be produced after distiller's grains fermentation. When distiller's grains were mixed with Zanthoxylum bungeanum leaves for fermentation, the ABTS scavenging ability was further enhanced, indicating that the mixed fermentation of distiller's grains and Zanthoxylum bungeanum leaves can promote the synthesis of antioxidant active substances.

[0072] Providing exogenous antioxidants can improve animal health, potentially reduce the need for antibiotic supplements, and provide alternative feeding options that can still maintain healthy and safe animal products. Zanthoxylum bungeanum leaves have many antioxidant active substances, so the antioxidant activity is high before fermentation. After fermentation, the anti-nutritional factors are reduced and some antioxidant active substances are also lost. Whether Zanthoxylum bungeanum leaves are fermented as raw materials alone or mixed with distiller's grains, the antioxidant activity is more retained after inoculation of mixed bacterial agents, and the nutritional value is higher. The increase in the retention rate of antioxidant activity may be because the metabolism of lactic acid bacteria inhibits the growth of miscellaneous bacteria, the degradation of antioxidant substances is reduced, and at the same time, microbial metabolism also produces some substances with antioxidant activity, such as peptides.

[0073] After Zanthoxylum bungeanum leaves are mixed with distiller's grains, their nutritional structure is more reasonable, suitable for the growth and fermentation of microorganisms, but not inoculating mixed bacterial agents will cause the growth of miscellaneous bacteria, increase the weight loss rate, and the nutritional structure is destroyed, and the antioxidant activity loss is more. The present invention adopts bacillus subtilis, saccharomyces cerevisiae and short growth-promoting lactobacillus and / or plant lactobacillus to prepare mixed bacterial agents, and by inoculating three kinds of mixed bacterial agents, the microbial community structure in the solid-state fermentation system is changed, the growth of harmful microorganisms is effectively controlled, the content of anti-nutritional factors is reduced, the antioxidant activity is improved, the DM loss is reduced, the soluble sugar content is increased, and the true protein has no obvious loss, which can provide animals with higher nutritional value. Wherein, embodiment 3 (short growth-promoting lactobacillus, plant lactobacillus, bacillus subtilis and saccharomyces cerevisiae) shows the most comprehensive and best solid-state fermentation performance, and its antioxidant activity can be more retained, and can replace antibiotics as animal feed additives, which is of positive significance for protecting animal health. The present invention is of great significance for the comprehensive recycling of Zanthoxylum bungeanum leaves and distiller's grains in the production of unconventional feed resources.

[0074] Table 1

[0075]

[0076]

[0077] Table 2

[0078]

[0079]

[0080] Table 3

[0081]

[0082] Table 4

[0083]

[0084] The dry weight retention rate DMR, tannin degradation rate, phytic acid degradation rate, α-sanshool degradation rate, antioxidant activity retention rate and true protein content TPN of Examples 3-5 are shown in Table 5. It can be seen that in Examples 4-5, the additives made of stachyose, trehalose, magnesium sulfate, manganese sulfate and diammonium hydrogen citrate are added to the fermentation substrate, which is more suitable for the growth and fermentation of the four microorganisms (Lactobacillus brevis, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae) of the present invention; in addition, in Example 4, the fermentation substrate is irradiated and pretreated before inoculation of the bacterial agent to make the fermentation substrate active, and the subsequent inoculation of the mixed bacterial agent can promote the metabolism of the microorganisms. The obtained Zanthoxylum leaf-vintner's lees fermentation composition has a higher retention rate of antioxidant activity, a higher degradation rate of tannin, phytic acid and α-sanshool, a higher DMR, and a reduced DM loss than that of Example 3.

[0085] Table 5

[0086]

[0087] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to the specific details and the embodiments shown and described herein.

Claims

1. A Zanthoxylum bungeanum leaf-vinasse fermentation composition, characterized in that: It includes the following raw materials: Wine lees, Zanthoxylum bungeanum leaves and mixed bacterial agents; The mixed bacterial agent comprises Levilactobacillus brevis, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae.

2. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 1, characterized in that: The dry weight ratio of the wine lees to the Zanthoxylum bungeanum leaves is 1:0.5-2; the dosage of the mixed bacterial agent is 5-15wt% of the total weight of the wine lees and the Zanthoxylum bungeanum leaves.

3. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 1, characterized in that: The Zanthoxylum bungeanum leaf-vintner's lees fermentation composition is obtained by subjecting the raw materials to anaerobic solid-state fermentation.

4. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 1, characterized in that: In the mixed bacterial agent, Lactobacillus breve, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae are composed in a mass ratio of 1-3:1-3:1-3:1-3.

5. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 1, characterized in that: The mixed bacterial agent comprises Lactobacillus breve, Bacillus subtilis and Saccharomyces cerevisiae.

6. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 5, characterized in that: In the mixed bacterial agent, Lactobacillus breve, Bacillus subtilis and Saccharomyces cerevisiae are composed in a mass ratio of 1-3:1-3:1-3.

7. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 1, characterized in that: The mixed bacterial agent is lactobacillus plantarum, bacillus subtilis and saccharomyces cerevisiae.

8. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 7, characterized in that: In the mixed bacterial agent, Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae are composed in a mass ratio of 1-3:1-3:1-3.

9. The Zanthoxylum bungeanum leaf-vinasse fermentation composition according to claim 3, characterized in that: The specific steps of the anaerobic solid-state fermentation are: crushing distiller's grains and Zanthoxylum leaves into 20-50 meshes and then mixing them evenly, then mixing them with a mixed bacterial agent, stirring evenly, adjusting the water content of the raw materials to 50-70wt%, and conducting sealed anaerobic fermentation for 4-6 days at a fermentation temperature of 35-40°C to obtain a Zanthoxylum leaf-distiller's grains fermentation composition.

10. Use of the Zanthoxylum bungeanum leaf-vinasse fermentation composition according to any one of claims 1 to 9 in an animal feed additive.

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