Preparation and application of apple-pomace-containing pet bacterium-enzyme synergistic fermentation feed

Abstract

The invention relates to the technical field of fermented feeds, in particular to a fermented feed adopting apple pomace as a main raw material for pets. The invention provides a preparation method and application of a pet bacterium-enzyme synergistic fermentation feed taking apple pomace as a main raw material. A formula of a fermentation substrate is innovated, pichia pastoris protein hydrolysate is used for replacing a traditional soybean meal nitrogen source, and a bacterium-enzyme synergistic fermentation technology is utilized, so that on one hand, fibers and anti-nutritional factors in raw materials are hydrolyzed by utilizing an enzyme preparation; on the other hand, beneficial metabolites are generated through microbial fermentation. The produced product has a relatively high dietary fiber level, a proper proportion of insoluble and soluble fibers, contains beneficial microbial metabolites, and has remarkable effects of relieving pet constipation, improving intestinal flora balance and improving intestinal health of pets.

Description

Technical Field

[0001] The invention relates to the technical field of fermented feed, in particular to a fermented feed for pets which adopts apple pomace as a main raw material. Technical Background

[0002] As more and more people keep pets, the pet feed industry is also expanding. Intestinal problems are one of the most common problems for pets. The intestine is not only the main place for the digestion and absorption of nutrients, but also an innate barrier to ensure the homeostasis of the body's internal environment. Food digestion relies on the power generated by the digestive tract and the participation of many substances and enzymes that help digestion. When food raw materials pass through the digestive tract, each part of the digestive tract performs specific and different digestive functions on it. When a pet's intestine has problems, its digestion and absorption function is impaired, affecting the absorption of nutrients, etc., threatening the pet's health and even life.

[0003] There are many factors that affect the intestinal function problems of pets, such as pathogenic factors, nutritional factors, poisoning factors, environmental factors, etc. Feeding pets with incomplete, unbalanced, and low-quality feed can easily cause nutritional deficiencies in the body, which will directly affect the function of the intestinal mucosal immune system and the health of the gastrointestinal tract. Although the factors that cause intestinal problems are different, they will all cause intestinal flora imbalance and intestinal dysfunction, causing indigestion in the stomach and intestines, and easily cause constipation, fecal odor and other problems (Lv Shaohua, Wu Guangming, Lv Shaojun. A health food for regulating the intestinal function of pet dogs and cats and its preparation method [P]. CN108323641A.2024-10-12.), which seriously affects the growth and development of pets and is also a headache for pet breeders and technicians.

[0004] Apple pomace is the residue left after fresh apples are crushed, squeezed and juiced. It is mainly composed of peel, core and residual pulp. Apple pomace is cheap, widely available and rich in dietary fiber. The dietary fiber content of dry apple pomace is as high as 58.8%. Studies have found that adding 10% dry apple pomace to the diet makes the experimental animals' defecation smoother, the amount of feces increases, the color becomes darker, and the incidence of constipation is significantly reduced (Deng Dun, Duan Yehui, Yao Kang, et al. Effects of adding apple pomace to the diet on feces and reproductive performance of pregnant sows [J]. Journal of Animal Nutrition, 2015, 27(2): 535-540.). Lack of crude fiber can cause problems in the body's defecation and digestive system. In addition, the dietary fiber level, that is, the ratio of insoluble to soluble fiber, has an important influence on relieving animal constipation and exerting reproductive potential (Wu De. Fiber nutrition is an important factor in fully exerting the reproductive potential of sows [J]. Northern Animal Husbandry, 2018, 561(17): 21.). Microbial fermentation technology can increase the soluble fiber content in apple pomace and improve its hydration properties (Qiu Jiying, Chen Leilei, Wang Weiming, et al. Research progress in the preparation of high-quality dietary fiber by fermentation [J]. Chinese Food and Nutrition, 2010, (6): 24-27.), which plays an important role in relieving pet constipation and improving intestinal problems.

[0005] CN106615810A discloses a fermented material made from corn, soybean meal, premix, bran, wet brewer's grains, corn germ meal, cornmeal, bean curd residue, animal skin fluid, stone powder, microbial fermentation agent and Chinese herbal medicine additive (made from jujube, Rehmannia root, wolfberry, astragalus and motherwort, mixed and ground into a dry powder preparation). The fermentation technology is used to degrade anti-nutritional factors in unconventional feed raw materials, improve the utilization rate of unconventional feeds such as brewer's grains, reduce feed costs, increase beneficial bacteria and enzyme activity in the animal intestine, degrade mycotoxins, and reduce the occurrence of diseases such as constipation.

[0006] CN104161210A discloses a feed raw material for pregnant sows, which is prepared by fermenting and enzymolyzing 400-600 parts of apple pomace, 4-6 parts of brown sugar, 4-6 parts of lysine and 0.4-0.6 parts of ferrous sulfate monohydrate, and then adding corn flour, soybean meal, soybean oil, calcium powder, calcium hydrogen phosphate, salt and additives to prepare feed for pregnant sows. Since the feed contains a large amount of crude fiber, probiotics, oligopeptides, lactic acid, vitamins, unknown growth factors and the like, it has the effects of improving the palatability of livestock and poultry, increasing the digestion and absorption rate of nutrients, promoting the growth of livestock and poultry, and preventing and treating constipation and diarrhea.

[0007] In recent years, with the vigorous development of the bio-feed industry, the bacterial enzyme synergistic fermentation technology has gradually evolved. The bacterial enzyme synergistic fermentation technology is designed based on the characteristics of the fermentation substrate, with a unique combination of microorganisms, substrates and enzymes, taking into account the microbial nutrition, animal nutrition and intestinal flora balance, and showing more outstanding technical advantages in improving animal health and production performance indicators. However, there are no reports on the use of bacterial enzyme synergistic technology to process apple pomace to produce fermented feed for pets. Summary of the invention

[0008] In view of the deficiencies in the prior art, the present invention provides a preparation method and application of a pet feed prepared by co-fermentation of fungi and enzymes with apple pomace as a main raw material.

[0009] The invention provides a method for preparing a bacterial enzyme synergistic fermented feed additive for pets, which comprises taking apple pomace as a main raw material, mixing a fermented raw material mixture in a certain proportion, then inoculating a mixed bacterial agent and a composite enzyme preparation, fully mixing and fermenting, and preparing the bacterial enzyme synergistic fermented feed for pets.

[0010] Among them, the mixed bacterial agent is composed of Bacillus and Saccharomyces kudriavzii.

[0011] Among them, the inoculum amount of bacteria is: 4~15×10 6 CFU / kg, Kudriavzia yeast 5~20×10 9 CFU / kg.

[0012] More specifically, the Bacillus is Bacillus subtilis YQ-XJ-2, which is commercially available, for example, from Beijing Baozang Biotechnology Co., Ltd.

[0013] Kudriavzevii yeast is Kudriavzevii yeast Kud-A strain, which is deposited in the General Microbiological Center of China Microorganism Culture Collection Administration (referred to as CGMCC, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing), with a deposit number of CGMCC NO.26899 and a deposit date of February 24, 2023. Its classification name is Pichia kudravzevii (and disclosed in Chinese patent application 2023107076301).

[0014] The complex enzyme preparation is fermented and produced by Trichoderma reesei A2H. The strain Trichoderma reesei A2H is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number: CGMCC No.21470, the deposit time: March 17, 2021, and the address of the deposit unit: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing (and disclosed in Chinese patent application 2023112060144).

[0015] The amount of enzyme preparation added is 0.03% to 0.05% of the dry matter of the fermentation raw material mixture.

[0016] The fermentation raw material mixture consists of the following components by weight percentage: 55% to 70% of apple pomace, 10% to 20% of rice bran meal, 10% to 15% of bran, and 10% to 15% of Pichia pastoris hydrolyzate.

[0017] Apple pomace, rice bran meal, bran and Pichia pastoris hydrolysate are crushed to a particle size of ≤2 mm as required.

[0018] Prepare a fermentation raw material mixture by mixing according to the above proportions, take an appropriate amount of fluoride-free water, dissolve the mixed bacterial agent and compound enzyme preparation in the water, and evenly spray the mixture to make the moisture content of the mixture 30% to 40%. After fully mixing, put it into a breathing bag and seal it, and ferment it at 25 to 30°C for 5 days.

[0019] The present invention also provides a fungus enzyme cooperatively fermented feed additive for pets obtained by the preparation method.

[0020] Further provided is a pet-use bacterial enzyme synergistic fermented feed, which is obtained by adding the feed additive to a feed ration, preferably, the added amount is 1-15%, preferably 2-10%, specifically 4-5%.

[0021] The present invention further provides the aforementioned microbial enzyme synergistically fermented feed additive for pets, or the use of the aforementioned microbial enzyme synergistically fermented feed for pets in raising pets.

[0022] Specifically, the pet is a pet fragrant pig; the breeding stage is the gestation period, especially the late gestation period; specifically, it is used to relieve pet constipation, improve the balance of intestinal flora and / or improve the intestinal health of pets.

[0023] The fermentation indexes and nutritional indexes of the pet bacterial enzyme synergistically fermented feed containing apple pomace provided by the present invention are shown in Table 1.

[0024] Table 1. Nutritional composition analysis

[0025]

[0026] The present invention uses an innovative fermentation substrate formula, replaces the traditional soybean meal nitrogen source with Pichia pastoris protein hydrolysate, and utilizes bacterial enzyme synergistic fermentation technology. On the one hand, enzyme preparations are used to hydrolyze the fiber and anti-nutritional factors in the raw materials; on the other hand, microbial fermentation is used to produce beneficial metabolites. The product produced has a high level of dietary fiber, a suitable ratio of insoluble to soluble fiber, and contains beneficial microbial metabolites, which has a significant effect on relieving pet constipation, improving the balance of intestinal flora, and improving the intestinal health of pets. DETAILED DESCRIPTION

[0027] The present invention is described below through specific implementation modes in order to better understand the present invention, but it does not constitute a limitation of the present invention.

[0028] Raw material preparation example

[0029] Preparation of Bacillus subtilis: Culture in LB medium (yeast extract 5.0 g / l, peptone 10.0 g / l, sodium chloride 10.0 g / l) at 37°C for 24 hours.

[0030] Preparation of Pichia kudriavzii KuA: Culture on YPD medium (peptone 20 g / l, yeast extract 10 g / l, glucose 20 g / l) at 30°C for 24 hours.

[0031] Preparation of compound enzyme preparation:

[0032] Trichoderma reesei A2H was cultured under the following conditions: culture temperature of 24°C to 28°C, pH 4.8 to 5.2, rotation speed of 250 to 300 rpm, dissolved oxygen content of 25 to 35% (v / v) in the fermentation broth, and cultured for 120 hours. The culture medium contained the following components: 20 to 30 g / L glucose, 2 to 6 g / L corn steep liquor powder, 1.60 to 1.72 g / L KOH, 2.6 to 3.0 g / L (NH4)2SO4, and 0.4 to 0.8 g / L MgSO4.

[0033] Preparation of other raw materials: Apple pomace, rice bran meal, bran and Pichia yeast hydrolysate are crushed to a particle size of ≤2 mm as required.

[0034] Example 1

[0035] This embodiment relates to a bacterial enzyme-assisted apple pomace fermented feed additive for improving pet constipation. According to weight percentage, 60% apple pomace, 10% rice bran meal, 15% bran and 15% Pichia pastoris hydrolyzate are mixed, and the mixed bacteria and compound enzyme are dissolved in an appropriate amount of water and sprayed on the mixture so that the moisture content of the mixture is 30% to 40%. After fully mixing, the mixture is sealed in a breathing bag and fermented at 25 to 30°C for 5 days. The addition amount of each bacteria in the mixed bacteria is: Bacillus subtilis YQ-XJ-24.0-6.4×10 6 CFU / kg, Pichia kudriavitzii 5.5×10 9 CFU / kg. The added amount of the complex enzyme was 0.04% of the dry matter of the fermentation raw material mixture.

[0036] The obtained pet enzyme synergistic fermentation feed additive has sour and wine aroma, no bad smell, is loose and soft without lumps, and the nutritional composition is shown in Table 2.

[0037] Table 2: Nutritional composition of bacterial enzyme-co-fermented feed additives for pets during pregnancy

[0038]

[0039]

[0040] Example 2

[0041] This embodiment relates to a bacterial enzyme-assisted apple pomace fermented feed additive for improving constipation in pets during pregnancy. According to the weight percentage, 70% apple pomace, 10% rice bran meal, 10% bran and 10% Pichia pastoris hydrolyzate are mixed, and the mixed bacteria and compound enzyme are dissolved in an appropriate amount of water and sprayed on the mixture so that the moisture content of the mixture is 30% to 40%. After fully mixing, the mixture is sealed in a breathing bag and fermented at 25 to 30°C for 5 days. The addition amount of each bacteria in the mixed bacteria is: The addition amount of each bacteria in the mixed bacteria is: Bacillus subtilis YQ-XJ-210×10 6 CFU / kg, Pichia kudriavitzii 3.2×10 7 CFU / kg. The added amount of the complex enzyme was 0.03% of the dry matter of the fermentation raw material mixture.

[0042] The obtained bacterial enzyme synergistic fermentation feed additive for pets has sour and wine aroma, no bad smell, is loose and soft without lumps, and the nutritional composition is shown in Table 3.

[0043] Table 3. Nutritional composition of bacterial enzyme synergistic fermentation feed additives for pets during pregnancy

[0044]

[0045] Example 3

[0046] This embodiment relates to a bacterial enzyme-assisted apple pomace fermented feed additive for improving constipation in pets during pregnancy. According to weight percentage, 55% apple pomace, 20% rice bran meal, 15% bran and 10% Pichia pastoris hydrolyzate are mixed, and the mixed bacteria and compound enzyme are dissolved in an appropriate amount of water and sprayed on the mixture, so that the moisture content of the mixture is 30% to 40%. After being fully mixed, the mixture is sealed in a breathing bag and fermented at 25 to 30°C for 5 days.

[0047] The amount of each bacteria added in the mixed bacteria is: Bacillus subtilis YQ-XJ-27.2×10 6 CFU / kg, Kudriavzia yeast 4.5×10 7 CFU / kg. The added amount of the complex enzyme is 0.05% of the dry matter of the fermentation raw material mixture.

[0048] The obtained bacterial enzyme synergistic fermentation feed additive for pets has sour aroma and wine aroma, no bad smell, and is loose, soft and free of lumps.

[0049] Test example

[0050] 45 late-pregnancy pet pigs with similar parity, body condition and good health were selected and randomly divided into 3 groups, with 15 replicates in each group and 1 pig in each replicate. The experimental pigs were fed with commercial early-pregnancy compound feed Antaibao (1-89 days) and late-pregnancy compound feed Santongbao (90-114 days), both purchased from Jiuzhou Dadi Feed Co., Ltd. Experimental Group 1 and Group 2 were additionally added with 5% and 10% of the fermented apple pomace feed prepared in Example 1 on the basis of commercial feed, respectively. The experimental pet pigs were kept in a limited fence, raised in a single fence, with restricted feeding and free drinking water. The pens were disinfected and cleaned regularly to keep the pens clean and airy. The test results are shown in Table 5. Adding 5% fermented apple pomace feed additive to the diet significantly reduced the incidence of constipation in fragrant pigs (P < 0.01), with a reduction of 39%; the number of piglets born alive, litter weight at birth and average weight of newborn individuals were also significantly improved (P < 0.05); the number of piglets born tended to increase (P> 0.05).

[0051] Table 5 Effects of adding apple pomace fermented feed additive to the diet on the incidence of constipation and reproductive performance of pregnant pet pigs

[0052]

[0053] Data in the same row with no letters or the same letters indicate no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

Claims

1. A method for preparing a bacterial enzyme-co-fermented feed additive for pets, characterized in that: A fermentation raw material mixture with apple pomace as the main raw material is then inoculated with a mixed bacterial agent and a compound enzyme preparation, and then fully mixed and fermented to prepare a bacterial and enzyme synergistic fermented feed for pets; Among them, the mixed bacterial agent is composed of Bacillus and Pichia kudriavzi; Trichoderma reesei Trichoderma reesei A2H fermentation production.

2. The preparation method according to claim 1, characterized in that The inoculum size of each bacterium is: 4~15×10 6 CFU / kg, Bacillus and Pichia kudriavitzii 5~20×10 9 CFU / kg; The addition amount of the complex enzyme preparation is 0.03%~0.05% of the dry matter of the fermentation raw material mixture.

3. The preparation method according to claim 1, characterized in that: The fermentation raw material mixture consists of the following components by weight: 55% to 70% apple pomace, 10% to 20% rice bran meal, 10% to 15% bran, and 10 to 15% Pichia pastoris hydrolyzate.

4. The preparation method according to claim 3, characterized in that: Apple pomace, rice bran meal, bran and Pichia pastoris hydrolysate were ground to a particle size of ≤2 mm as required.

5. The preparation method according to claim 1, characterized in that: Prepare a fermentation raw material mixture by mixing in proportion, take an appropriate amount of fluoride-free water, dissolve the mixed bacterial agent and compound enzyme preparation in the water, and evenly spray the mixture so that the moisture content of the mixture is 30%~40%; After fully mixing, put it into a breathing bag, seal it, and ferment it at 25-30℃ for 5 days to prepare it as a feed additive.

6. The fungus-enzyme synergistic fermentation feed additive for pets obtained by the preparation method according to any one of claims 1 to 5.

7. A bacterial enzyme synergistically fermented feed for pets, obtained by adding the feed additive according to claim 6 to a feed diet, preferably, the added amount is 1-15%, preferably 2-10%, specifically 4-5%.

8. Use of the bacterial enzyme synergistically fermented feed additive for pets as claimed in claim 6, or the bacterial enzyme synergistically fermented feed for pets as claimed in claim 7 or 8 in raising pets.

9. The use according to claim 7, characterized in that The pet is a pet fragrant pig; the breeding stage is the gestation period, especially the late gestation period.

10. The use according to claim 9, characterized in that Used to relieve pet constipation, improve intestinal flora balance and / or improve pet intestinal health.

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