Lactobacillus plantarum capable of improving animal intestinal health, culture, microbial inoculum and application of lactobacillus plantarum

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum capable of improving animal intestinal health, a culture, a fungicide and application of the lactobacillus plantarum. The lactobacillus plantarum QZF is preserved in the China General Microbiological Culture Collection Center (CGMCC) on July 22, 2024, and the preservation number is CGMCC No. 31389. The lactobacillus plantarum QZF has the advantages that the lactobacillus plantarum QZF is preserved in the China General Microbiological Culture Collection Center; according to the invention, a microecological preparation based on lactobacillus plantarum is created, and a solid / liquid state fermentation mode suitable for pig farms is researched and developed, so that the intestinal health of animals is improved, the efficient utilization of feed resources is promoted, and the breeding benefits are improved; when the feed additive is applied to pregnant sows, the intestinal health degree of the sows can be improved, the birth process can be shortened, meanwhile, the intestinal health of newly born piglets is improved in a mother-child transfer mode, the young piglets are assisted to establish balanced intestinal flora, and the weaning stress of the piglets is reduced.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbiology, and particularly relates to a Lactobacillus plantarum strain capable of improving animal intestinal health, a culture, a microbial agent and their applications. Background Art

[0002] The development of the innate immune function of the intestinal mucosa in neonatal animals plays a key role in their healthy growth and is closely related to the intestinal flora. Intestinal diseases are one of the main causes of morbidity and mortality in neonatal animals such as weaned piglets, weaned meat rabbits, calves, etc. Improving intestinal health is an important means to increase the survival rate of neonatal animals and improve the efficiency of aquaculture production. The imperfect development of the digestive tract and the imperfect intestinal immune function lead to neonatal animals being easily infected with pathogenic bacteria, thus causing a series of diseases. The gastrointestinal tract is not only a place for nutrient absorption and digestion, but also a colonization site for complex, diverse and dynamic microbial communities, and these microorganisms are constantly "exchanging information" with the host. During the critical window of early life development, neonates can regulate the development of the immune system through specific signals from microorganisms and increase their resistance to certain diseases.

[0003] Previous studies by many researchers have proven that intestinal microorganisms can indeed help and maintain metabolic functions, can affect the development and function of the immune system, and have important significance for the healthy growth and development of the host; the intestinal flora of human neonates can promote the innate and adaptive immune development of germ-free animals such as germ-free mice and reduce intestinal inflammatory responses after extraction, isolation and transplantation. The immune system of neonatal animals is not perfectly developed. Taking neonatal piglets as an example, they need to rely on immune factors in breast milk to prevent the colonization and overgrowth of pathogenic bacteria. Intestinal microorganisms can help the host resist pathogenic bacteria by regulating the intestinal immune system. For example, segmented filamentous bacteria are known inducers of the animal intestinal immune system and are also dominant species in the ileal flora community of weaned piglets. Intervention in the early life of animals helps neonatal animals quickly establish a good intestinal microbiota, which has a promoting effect on the later growth, immune system development and healthy growth of neonatal animals. Direct supplementation of probiotics is one of the means that can effectively improve the composition of the intestinal microbiota in the early stage of life. Supplementing lactobacilli to newly born piglets can improve the intestinal health of piglets after weaning, increase the abundances of lactobacilli and bifidobacteria, and reduce the numbers of potential intestinal pathogenic bacteria such as Escherichia coli ( Escherichia coli ), and Clostridium ( Clostridium ). Randomized double-blind clinical trials in human medical research have also found that administering probiotic mixtures to infants after birth can significantly improve the species and functional composition of the microbiota. Therefore, developing new composite microecological agents with probiotic characteristics can increase the survival rate of neonatal animals, improve the efficiency of aquaculture production, achieve "cost reduction and efficiency increase", and create economic value.

[0004] At present, in the era of "antibiotic ban", Lactobacillus plantarum, as a kind of probiotic, has become a hot research direction in the field of green feed additives due to its many advantages. However, although the research and application of Lactobacillus plantarum are increasingly valued, the problem of low live bacteria count in Lactobacillus plantarum products still exists in practical applications. Summary of the invention

[0005] In order to solve the problem that the existing plant lactobacillus in the prior art still has a low number of viable bacteria in the plant lactobacillus product in practical application, the present invention proposes a plant lactobacillus that can improve the intestinal health of animals, a culture and a bacterial agent and the application thereof. To achieve the above object, the present invention adopts the following technical scheme.

[0006] The present invention provides a strain of Lactobacillus plantarum QZF that can improve the intestinal health of animals. The Lactobacillus plantarum QZF was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration on July 22, 2024, with a deposit number of CGMCC No. 31389. The Lactobacillus plantarum QZF is also known as Lactobacillus plantarum. L. plantarum QZF.

[0007] In the prior art, when Lactobacillus plantarum is actually applied, it is often made into dry powder and then directly mixed into feed according to proportion for addition. However, since the Lactobacillus plantarum strain is in a dormant stage, it needs to be activated in the animal body before it can exert a probiotic effect, so the effect is uneven and directly related to the technical method of producing dry powder. Therefore, the existing Lactobacillus plantarum still has the problem of low viable count in Lactobacillus plantarum products in practical applications. The present invention provides a strain of Lactobacillus plantarum QZF that can improve the intestinal health of animals and its field application form, which can ensure that the animal can ingest a sufficient amount of live bacteria during production, and can also ensure the various beneficial metabolites produced by active probiotics; the pH of the feed after fermentation is low, which can inhibit the growth of harmful bacteria and has a sour aroma, improve animal appetite, and increase feed intake; compared with traditional technology, the probiotic effect of Lactobacillus plantarum can be significantly improved, and the problem of low viable count in Lactobacillus plantarum products in practical applications of the existing Lactobacillus plantarum in the prior art is solved.

[0008] Through the solid and liquid culture medium and on-site fermentation technology of the present invention, it can be ensured that the probiotics are in an active state when applied to animal production, and the content of various beneficial metabolites (such as lactic acid, indoleacetic acid, etc.) produced by the live bacteria can also be guaranteed. The pH of the fermented feed is low, which can inhibit the growth of harmful bacteria and has a sour aroma, improves the appetite of animals, and increases feed intake. These are advantages that traditional technologies cannot match.

[0009] The plant lactobacillus QZF provided by the present invention can enhance the balance of intestinal microorganisms, and effectively promote the proliferation of beneficial flora in the intestine by competitively inhibiting the growth of harmful flora, thereby significantly improving and maintaining the microbial ecological balance of the animal intestine, especially for alleviating the intestinal flora imbalance caused by stress conditions such as weaning. It has a significant effect; this strain of lactobacillus can stimulate the intestinal mucosal immune system of animals, enhance the activity of immune cells, and promote the production of immunoglobulins, thereby significantly improving the resistance of animals to intestinal pathogens. This mechanism of action is of great significance for the prevention and treatment of intestinal infectious diseases, and solves the problem of insufficient improvement of animal intestinal immunity in the prior art. At the same time, this strain of lactobacillus can promote the secretion of digestive enzymes in animals and improve the digestibility of complex carbohydrates, proteins and fats in feed. This characteristic helps to improve the growth performance of animals, especially in improving feed conversion rate and reducing breeding costs, and is superior to many similar strains widely used in the current market.

[0010] The present invention also provides application of the Lactobacillus plantarum QZF in regulating the intestinal health of animals.

[0011] Preferably, the plant lactobacillus QZF is used to promote animal feeding. After adding the plant lactobacillus QZF to the feed, the animal's feed intake can be significantly increased, especially in response to stressful environments such as weaning, the strain (plant lactobacillus QZF that can improve the intestinal health of animals) can help animals maintain a high appetite level and improve feed conversion rate; long-term application of the strain in feed, animals show better growth performance, fast weight gain, reduced diarrhea rate, and good health.

[0012] The present invention also provides a culture comprising the Lactobacillus plantarum QZF.

[0013] The present invention also provides a bacterial agent for improving the intestinal health of animals, wherein the bacterial agent comprises the culture and the culture comprises the bacterial liquid of the Lactobacillus plantarum QZF.

[0014] The bacterial agent is composed of the culture of the plant lactobacillus QZF and glucose, and the plant lactobacillus QZF is fermented anaerobicly until the number of viable bacteria reaches 1×10 8 After the CFU / mL, 10 kg of wheat bran was mixed with every 100 L of bacterial solution, and freeze-dried. After freeze-drying, 100 g of glucose was mixed with every 400 g of bacterial dry powder to obtain the target bacterial agent. The target bacterial agent is easy to use and has obvious effects. While significantly maintaining activity, it can also improve the palatability of feed.

[0015] Preferably, the culture is a solid fermentation culture or a liquid fermentation culture. By optimizing the conditions, the solid fermentation culture or the liquid fermentation culture can maintain the activity of the strain (the Lactobacillus plantarum QZF), ensuring that it plays the best effect in the process of feed addition or animal feeding. Both forms of culture are easy to handle and store, and are convenient for application in actual production. The application of the fermentation culture of the Lactobacillus plantarum QZF can effectively play its role in promoting the intestinal health of animals. This solves the problem that the existing Lactobacillus plantarum products in the prior art still have low viable counts, easy inactivation of strains, and unstable effects in practical applications. Solid fermentation culture or liquid fermentation culture can significantly improve the intestinal health level of animals, which is manifested as a more balanced intestinal microbial community, enhanced intestinal immunity, and improved nutrient absorption efficiency; it can stimulate the appetite of animals, increase feed intake, thereby promoting the improvement of growth performance, and maintaining a healthy intestinal environment, improving animal health and production efficiency.

[0016] Preferably, the culture comprises a bacterial liquid of the Lactobacillus plantarum QZF.

[0017] The bacterial liquid is a fermentation liquid obtained by inoculating the plant lactobacillus QZF into a separation and purification culture medium and fermenting the fermentation liquid.

[0018] The separation and purification culture medium includes any one of LB solid culture medium, LB liquid culture medium and MRS broth culture medium. Among the above three culture mediums, LB solid culture medium is suitable for the preliminary separation and purification of the plant lactobacillus QZF, and the solid form is convenient for observing the colony morphology for preliminary screening; LB liquid culture medium is used for the amplification and cultivation of the plant lactobacillus QZF, and provides enough bacterial volume for subsequent purification and identification; MRS broth culture medium is particularly suitable for the separation and purification and cultivation of the plant lactobacillus QZF, and the nutrients required for the growth of the plant lactobacillus QZF are particularly added to its composition, such as glucose, mannitol, ammonium citrate, etc., which can more effectively support the growth and reproduction of the plant lactobacillus QZF, and is used for a large number of expansions. According to the situation, the production efficiency of the plant lactobacillus QZF can be significantly improved, and production costs can be reduced.

[0019] Preferably, the bacterial agent is processed into an agriculturally acceptable preparation.

[0020] The dosage form of the bacterial agent is any one of liquid, powder and granule.

[0021] The present invention also provides the use of the culture or the bacterial agent in the preparation of a product that improves the intestinal health of animals, and the product includes any one or more of fermented feed, preservatives, silage additives, and livestock and poultry intestinal flora stabilizers. The culture or bacterial agent of the plant lactobacillus QZF is applied to the feed fermentation process, which can effectively improve the nutritional value and digestibility of the feed, and at the same time produce beneficial metabolites, such as lactic acid, acetic acid, etc., to adjust the pH value of the animal intestinal tract, inhibit the growth of harmful bacteria, and promote the proliferation of beneficial bacteria, thereby improving the intestinal health level of the animal. The culture or bacterial agent of the plant lactobacillus QZF has a natural antiseptic effect. Due to the metabolites such as lactic acid produced during growth, the pH is reduced, the growth of microorganisms in food or feed is inhibited, and the shelf life is extended. In addition, the culture or bacterial agent of the plant lactobacillus QZF can also be used as a silage additive to promote the fermentation process of silage, improve the quality and nutritional value of feed, inhibit the growth of harmful microorganisms during silage, and reduce the accumulation of undesirable substances in feed, thereby improving the intestinal environment of animals. The culture or bacterial agent of Lactobacillus plantarum QZF can be used as a stabilizer for the intestinal flora of livestock and poultry, and can be added into feed to directly act on the intestinal tract of animals, regulate the balance of intestinal flora, enhance intestinal immunity, and improve the resistance of animals to diseases.

[0022] The present invention also provides a product for improving the intestinal health of animals, comprising the culture of the Lactobacillus plantarum QZF or the bacterial agent, and an auxiliary material acceptable to food and / or health products.

[0023] The present invention separates plant lactobacillus from the fermentation liquid of giant fungus grass and is temporarily named Lactobacillus plants QZF, referred to as L. plantarum QZF. In the previous study, it was found that the fermented liquid of Lactobacillus plantarum has a sour and fragrant smell, which can promote animal feeding. Based on this, the present invention is based on L. plantarum Taking QZF as the starting point, with the help of solid / liquid fermentation technology and animal experiments, we systematically studied the effects of probiotic fermentation on animal reproductive performance and changes in intestinal flora composition, and analyzed the probiotic mechanism, providing data support for the further development of complex and efficient microecological preparations and their application in animal husbandry production, in order to provide the market with high-quality animal husbandry products and new microecological preparations.

[0024] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention provides a strain of Lactobacillus plantarum that can improve the intestinal health of animals. The Lactobacillus plantarum QZF that can improve the intestinal health of animals provided by the present invention was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration Committee on July 22, 2024, and the deposit number is CGMCC No. 31389. The Lactobacillus plantarum QZF that can improve the intestinal health of animals provided by the present invention and its field application form can ensure that animals can ingest sufficient live bacteria during production, and at the same time can also ensure the various beneficial metabolites produced by active probiotics; the pH of the fermented feed is low, which can inhibit the growth of harmful bacteria and has a sour aroma, improve animal appetite, and increase feed intake; compared with traditional technologies, the probiotic effect of Lactobacillus plantarum can be significantly improved, and the problem that the number of live bacteria in Lactobacillus plantarum products in the existing Lactobacillus plantarum products in practical applications in the prior art is still low.

[0025] 2. The plant lactobacillus QZF that can improve the intestinal health of animals provided by the present invention can be applied to the diet for pigs. It has a prebiotic effect, can not only improve the quality of pork, but also promote the growth and development of piglets, can also alleviate the related symptoms of weaned piglets infected with Escherichia coli, inhibit diarrhea, improve production performance, and improve intestinal health to improve the survival rate of newborn animals and improve the production efficiency of breeding. It is also possible to improve the intestinal damage caused by weaning of newborn animals by improving intestinal barrier function and antioxidant activity. In addition, plant lactobacillus also helps to relieve diarrhea, prevent colitis, regulate the structure of animal intestinal flora, reduce the risk of food production and animal disease, and plant lactobacillus can effectively inhibit fumonisin in feed. Adding plant lactobacillus to the diet can improve the utilization rate of feed by animals, while improving stress resistance and immunity. In the application of food field, probiotic plant strains usually appear in the food industry as probiotics or microbial fermentation agents. It can be applied in cheese production, and plant lactobacillus can improve the flavor, texture and storage time of cheese.

[0026] 3. The present invention creates a microecological preparation based on plant lactobacillus, and develops a solid / liquid fermentation model suitable for pig farms, which improves the intestinal health of animals, promotes the efficient use of feed resources, and improves breeding efficiency; its application in pregnant sows can improve the intestinal health of sows, shorten the delivery process, and at the same time improve the intestinal health of newborn piglets through mother-to-child transmission, assist young piglets in establishing a balanced intestinal flora, and reduce weaning stress in piglets.

[0027] At the same time, the successful development and application of corresponding microecological preparations can improve animal intestinal health, increase feed digestibility, reduce the use of antibiotics, and to a certain extent reduce the production of gases such as ammonia and hydrogen sulfide in livestock farms, thereby alleviating the environmental pressure brought about by intensive animal husbandry in our province. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The single bacterial colony obtained by streaking and the fermented feed obtained in the present invention; wherein, Figure 1 Figure A in the figure is a single colony obtained by streaking in the present invention, and Figures 1 and 2 are plates obtained by QZF streaking, which are the results obtained by parallel experiments; Figure 1 Figure B is a demonstration diagram of a fermentation tank on site, and Figures 1 and 2 are the results of parallel tests; Figure 1 Figure C is a demonstration of the on-site fermentation effect.

[0029] Figure 2 This is the sequence alignment in the present invention.

[0030] Figure 3 The jejunum gene Q-PCR result obtained in Example 2 of the present invention; wherein, Figure 3 Panel A in the figure shows the Q-PCR result of IL-4 gene; Figure 3 Figure B is the Q-PCR result of IFN-γ gene; among them, Contrl is the control group; QZF is the Lactobacillus plantarum group; QZF+EMS is Lactobacillus plantarum + yeast; QZF+BD is Lactobacillus plantarum + spores.

[0031] Figure 4 The relative changes of the total number of piglets born + the number of live piglets and the relative changes of the number of mummies + the number of dead fetuses obtained in Example 3 of the present invention; wherein, Figure 4 Figure A shows the relative changes in total litter size; Figure 4 Figure B shows the relative change of live piglets; D-Contrl is the control group; B is the Lactobacillus plantarum group; C is Lactobacillus plantarum + spores; Figure 4 Figure C shows the relative changes in the number of mummies; Figure 4 Figure D shows the relative changes in the number of dead fetuses; among them, D-Contrl is the control group; B is the Lactobacillus plantarum group; C is Lactobacillus plantarum + spores.

[0032] Figure 5 The growth performance data of piglets before weaning obtained in Example 3 of the present invention; wherein, Figure 5 Figure A shows the average weight of piglets at birth; Figure 5 Figure B shows the average weight of piglets on the 7th day after birth; Figure 5 Figure C shows the average weight of piglets on the 14th day after birth; Figure 5 Figure D shows the average weight of piglets at weaning; among them, D-Contrl is the control group; B is the Lactobacillus plantarum group; and C is Lactobacillus plantarum + spores.

[0033] Figure 6The diarrhea rate data of piglet farrowing room obtained in Example 3 of the present invention; wherein, D-Contrl is the control group; B is the Lactobacillus plantarum group; and C is Lactobacillus plantarum + spores. DETAILED DESCRIPTION

[0034] The following will be combined with the drawings and specific embodiments of the present invention to clearly and completely describe the technical solutions in the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] The methods described in the embodiments of the present invention are conventional methods unless otherwise specified. The materials, reagents, etc. used can be obtained from commercial sources unless otherwise specified.

[0036] The culture medium and reagents used in the following examples are as follows: (1) Culture medium The formula of LB liquid culture medium is: 10 g / L trypsin, 10 g / L sodium chloride, 5 g / L yeast extract, and the solvent is water.

[0037] The formula of LB solid culture medium is: 10 g / L trypsin, 10 g / L sodium chloride, 5 g / L yeast extract, 1.5% / L agar powder, and the solvent is water.

[0038] The formula of the bacterial culture medium is: 10 g / L trypsin, 10 g / L sodium chloride, 5 g / L yeast extract, 2 g / L glucose, and the solvent is water.

[0039] The prepared bacterial culture medium is used to cultivate Lactobacillus plantarum. The specific method of use is: add a single colony of Lactobacillus plantarum to the bacterial culture medium and ferment it overnight at 37°C. Wherein, overnight means ≥12h. The Lactobacillus plantarum here refers to Lactobacillus plantarum QZF that can improve the intestinal health of animals.

[0040] The seed liquid culture medium is obtained by mixing 2%wt feed powder+2%wt glucose+94wt% water+2wt% bacterial liquid.

[0041] The formula of 300mL MRS broth medium is: 16.2g MRS, 4.5g agar powder, and the solvent is water.

[0042] (2) Reagents MRS broth culture medium was purchased from Shanghai Liquid Quality Testing; the inorganic salt reagents in the above culture medium were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0043] The culture dishes were purchased from Shanghai Liquid Quality Testing Co., Ltd.

[0044] 15mL centrifuge tubes were purchased from Shanghai Liquid Quality Testing; 1.5mL centrifuge tubes were purchased from Shanghai Liquid Quality Testing; 50mL centrifuge tubes were purchased from Shanghai Liquid Quality Testing.

[0045] Example 1: Obtaining Lactobacillus plantarum that can improve intestinal health of animals 1. Isolation and purification of strains After squeezing, the moisture content of freshly cut giant grass is controlled at 65%. After anaerobic fermentation for 15 days, 1g of fermented giant grass is taken out, 10mL of sterile water is added and mixed, and then spread on MRS broth medium plates. The MRS broth medium plates are placed in an anaerobic gas production bag and an anaerobic box, and incubated at 37℃ overnight. Overnight means ≥12h.

[0046] A strain of anaerobic bacteria with small and smooth colonies was picked from the MRS broth medium plate and temporarily named as anaerobic bacteria A1. After the anaerobic bacteria A1 was isolated, it was purified and cultured: a single colony was picked and cultured in MRS broth medium, and liquid paraffin was added to the MRS broth medium for liquid sealing, and cultured at 37°C overnight. This step is called activation.

[0047] The activated anaerobic bacteria A1 were prepared by mixing the bacterial solution with 40% by volume glycerol in a volume ratio of 1:1, and 40% by volume glycerol was added to the bacterial solution and mixed, and then the mixture was stored at -80°C.

[0048] 2. Identification of Lactobacillus plantarum that can improve animal intestinal health 2.1. Observation on the morphology and culture characteristics of Lactobacillus plantarum that can improve the intestinal health of animals After the above-obtained single colony of anaerobic bacteria A1 was expanded, it was repeatedly streaked on an MRS broth medium plate and anaerobically cultured for 24 hours. The plate colony was observed and it was found that the probiotic strain was spherical in shape. After the anaerobic bacteria A1 was revived, liquid paraffin was activated and placed in a 37°C incubator for anaerobically overnight. It was found that the fermentation liquid had a sour aroma. Among them, overnight means ≥12 hours.

[0049] 2.2 Sequencing and analysis of the 16S rRNA gene of Lactobacillus plantarum that can improve animal intestinal health (1) The genomic DNA of the anaerobic bacteria A1 was isolated and extracted using the MiniBEST Bacteria Genomic DNA Extraction Kit, which was purchased from Takara Bio. The extraction steps were performed according to the instructions of the kit. After obtaining the genomic DNA, 2×Hieff Canace Advancefast PCR Master Mix was used for amplification.

[0050] Among them, Hieff Canace Advancefast PCR Master Mix was purchased from Yishen Biotechnology (Shanghai) Co., Ltd.

[0051] During the amplification process, 16s rRNA universal sequencing primers 27F and 1492R were used. The specific primer sequences are shown below:

[0052] The nucleotide sequence of 27F is shown in SEQ ID NO. 2: 5'-AGAGTTTGATCCTGGCTCA-3'.

[0053] The nucleotide sequence of 1492R is shown in SEQ ID NO. 3: 5'-GGTTACCTTGTTACGACTT-3'.

[0054] The amplification program was as follows: 98°C: 30 s, 94°C: 10 s, 60°C: 5 s, 72°C: 5-10 s / kb, 30 cycles, 72°C: 2 min.

[0055] (2) After obtaining the PCR product, it was sent to Sangon Biotech (Shanghai) Co., Ltd. for Sanger second-generation sequencing. The sequencing result is shown in SEQ ID NO. 1:

[0056] The BLAST function on the NCBI website was used to align the sequences. The alignment results are shown in Figure 2 As shown. According to the comparison results, the anaerobic bacteria A1 was determined to be Lactobacillus plantarum. The URL of the NCBI website is: https: / / blast.ncbi.nlm.nih.gov / Blast.cgi.

[0057] Based on the morphological characteristics and sequencing analysis results, anaerobic bacteria A1 was preliminarily identified as Lactobacillus plantarum. Lactobacillus plantarum QZF, referred to as L. plantarum QZF. The plant lactobacillus QZF strain has been deposited at the General Microbiology Center of the China Microbiological Culture Collection Administration. The deposit address of the plant lactobacillus QZF strain is the Graduate School of Microbiology of the Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Its deposit date is July 22, 2024, and its deposit number is CGMCC No. 31389.

[0058] Among them, the plant lactobacillus QZF strain is also known as plant lactobacillus that can improve animal intestinal health L. plants QZF strain, Lactobacillus plantarum L. plantarum QZF. Hereinafter referred to as L. plantarum QZF.

[0059] Example 2: Research and application of solid / liquid fermentation mechanism of multi-strain combination 1. Test methods 1. Experimental Grouping 96 rabbits with similar body weight after weaning were randomly selected and divided into 4 groups, including a control group and 3 treatment groups. Each group had 4 replicates and each replicate had 6 rabbits. The control group was fed a basic diet.

[0060] All three treatment groups were added with a water-to-material ratio of 1:1.5. L. plantarum QZF. The seed solutions in the three treatment groups are group B: L. plantarum QZF, C: L. plantarum QZF+Yeast, Group D: L. plantarum QZF+ Bacillus subtilis s. Among them, Yeast is brewer's yeast, which was purchased from Xiamen Huiying Animal Technology Co., Ltd. Bacillus subtilis The source of s was purchased from Xiamen Huiying Animal Technology Co., Ltd.

[0061] The seed liquid of different treatment groups prepared in the above percentage was added to the basic diet for fermentation, and fermented feed of different treatment groups was obtained after 24 hours of fermentation. The mass percentage of the added seed liquid relative to the basic diet was 4%.

[0062] The rabbits in the above different treatment groups were fed with fermented feed obtained from different treatment groups. At the same time, the control group was fed with only the basic diet. The experimental period was 67 days, including a 7-day pre-feeding period and a 60-day feeding period. During the experiment, the rabbits were fed once in the morning and once in the evening, with routine feeding management and immunization, natural lighting and ventilation. After the experiment, the rabbits were slaughtered and samples were collected.

[0063] The rabbits are New Zealand white rabbits. Rabbits that have been weaned are also called weaned rabbits.

[0064] The amount of basal diet added was 100 g basal diet per rabbit. The basal diet was purchased from Fujian Chunlong Agriculture and Animal Husbandry Technology Co., Ltd.

[0065] The preparation methods of the fermented feed obtained by the above different treatment groups are the same except that the seed liquid added is different. L. plantarum The preparation method of fermented feed with QZF seed liquid fermented for 24 hours is taken as an example. The specific steps are as follows:

[0066] Resuscitation from -80°C L. plantarum QZF was activated by adding liquid paraffin and placed in a 37°C incubator for anaerobic incubation overnight. The plate was streaked onto an MRS plate, and the MRS plate was placed in an anaerobic gas production bag and an anaerobic box in a 37°C incubator overnight. L. plantarum QZF single colony was placed in liquid MRS, and then liquid paraffin was added and cultured anaerobically overnight at 37°C to obtain L. plantarum QZF activated bacterial solution. Overnight means ≥12h.

[0067] Further, the seed solution is prepared. The formula composition of the seed solution is: 2% by mass feed powder + 2% by mass glucose + 94% by mass water + 2% by mass L. plantarum QZF activated bacterial solution.

[0068] The prepared seed solution was cultured overnight at 37°C and 180 rpm in a shaking incubator.

[0069] The seed liquid was added to the basic diet at a mass ratio of 4% to the basic diet. After mixing well, the fermentation barrel was sealed and the lid was sealed. After anaerobic fermentation for 24 hours, the L. plantarum QZF fermented feed.

[0070] Among them, the water-to-feed ratio of water added during fermentation to the basic diet is 1:1.5.

[0071] The feeds fed to the different treatment groups and the control group are as follows: I. Blank group, also known as control group: 100g basic diet per rabbit.

[0072] II. Experimental group, also known as treatment group: (67g water + 100g feed per rabbit) * 16 rabbits * (1 + 4% seed solution), then 1.072kg water + 1.6kg feed + 107g seed solution is needed every day, and the total feed weight is 2.778kg. For 3 days, 3.2kg water + 4.8kg feed + 321g seed solution (6.5g feed powder + 6.5g glucose + 6.5g bacterial solution + 302g water) is needed, and the total feed weight is 8.424kg.

[0073] 2. Detection indicators ① Body weight and food intake Before and after the experiment, the weaned rabbits were weighed on an empty stomach. The weaned rabbits were weighed at 28, 42, and 70 days after the experiment and the average daily weight gain was calculated. The provided and remaining feed was collected and weighed daily to calculate the daily feed intake. The health of the weaned rabbits was observed daily, and the incidence and mortality were recorded. The experimental animals were weighed before and after slaughter, and the live weight and carcass weight before slaughter were recorded.

[0074] ②Intestinal length and contents After weaned rabbits were slaughtered, the lengths of the small intestine, cecum, and colorectum were measured, and the length / weight ratio was calculated based on the live weight before slaughter; the pH of the intestinal contents was measured, and the concentration of volatile fatty acids was determined by gas chromatography.

[0075] The steps of determining the concentration of volatile fatty acids by gas chromatography are as follows: Take 0.5g sample and grind it in liquid nitrogen, then add 4mL chloroacetic acid methanol, 1mL n-hexane and 1mL internal standard fatty acid solution. Then vortex the sample for 1 minute and keep it in a 75C water bath for 2 hours. Among them, the internal standard fatty acid solution is 1mg / mL 11-carbon fatty acid methyl ester. The English abbreviation of 11-carbon fatty acid methyl ester is FAME.

[0076] After cooling, 5 mL of 70 g / L potassium carbonate solution was added to the sample, vortexed for 1 min, and then centrifuged at 1200 rpm for 10 min. The supernatant was then loaded onto a gas chromatograph. The gas chromatography system was equipped with a KB-FFAP column and operated at 250 °C with a flame ionization detector. A N2 mobile phase with a flow rate of 0.81 mL / min was applied to the column. The column was operated at 80 °C. KB-FFAP column: Kromat, Delran, NJ, USA.

[0077] The concentration of a single fatty acid is calculated according to the following formula: Ci = (m0 * Ai * Fi * Ri) / (A0 * m), where Ci is the concentration of a single fatty acid (mg / g); m0 is the mass of the internal standard fatty acid (mg); m is the sample mass; Ai is the peak area of a single fatty acid in the sample; A0 is the peak area of the internal standard fatty acid; Fi is the correction factor from fatty acid methyl ester (FAME) to fatty acid (FA); Ri is the correction factor of the peak area.

[0078] ③ Histomorphological observation The histomorphology of the duodenum, jejunum, and cecum fixed with 4% paraformaldehyde was observed using HE staining. The villus height (C) and crypt depth (V) of the intestine were measured, and C / V was calculated. The intestinal morphology observation and tissue paraffin section were entrusted to Wuhan Sevier Biotechnology Co., Ltd. for sectioning and measurement.

[0079] ④ Determination of digestive enzyme activity The chyme of the duodenum, jejunum, and cecum was collected, and the enzyme activities of trypsin, pancreatic lipase, pancreatic amylase, and fibrolysin were determined using an ELISA kit.

[0080] Among them, the ELISA kits for detecting trypsin, pancreatic lipase, pancreatic amylase, and fibrolysin were purchased from Yeasen Biotechnology (Shanghai) Co., Ltd., and the determination steps were carried out according to the kit instructions.

[0081] ⑤ Determination of gastrointestinal hormones The jejunum tissue was collected, and the secretion amounts of gastrointestinal hormones such as CCK, GLP-1, and PYY, as well as the gene expression levels of ZO-1, occludin, claudin, TNF-α, IFN-r, IL-1β, IL-4, etc. were determined using ELISA or qPCR methods.

[0082] Among them, the steps for determining the secretion amounts of gastrointestinal hormones such as CCK, GLP-1, and PYY are as follows: After collecting 1 g of jejunum tissue and grinding it in liquid nitrogen, the secretion amounts of gastrointestinal hormones CCK, GLP-1, and PYY were determined using an enzyme-linked immunosorbent assay kit. The ELISA kit was purchased from Yeasen Biotechnology (Shanghai) Co., Ltd., and the determination steps were carried out according to the kit instructions. Among them, the English name of enzyme-linked immunosorbent assay is Enzyme-linked immunosorbent assay, ELISA.

[0083] The steps for determining the gene expression levels of occludin, claudin, TNF-α, IFN-r, IL-1β, IL-4, etc. are as follows: The RNA of jejunal tissue was extracted by TRIzol method, and the extraction steps were carried out according to the instruction manual of TRIzol. The cDNA reverse transcription kit Hifair AdvanceFast 1 st The extracted RNA was reverse transcribed using the Strand cDNA Synthesis Kit to obtain a cDNA library. The kit was purchased from Yisheng Biotechnology (Shanghai) Co., Ltd. and the procedure was performed according to the kit instructions.

[0084] The relative expression levels of gastrointestinal hormones occludin, claudin, TNF-α, IFN-r, IL-1β, and IL-4 mRNA in the jejunum were detected by qRT-PCR. -ΔΔ The Ct method was used to calculate the relative expression level of target gene mRNA.

[0085] The qRT-PCR primer design is shown in Table 1.

[0086] Table 1 qRT-PCR primer information ⑥ Blood index determination 4 mL of fasting heart blood was collected for the determination of blood indicators. ELISA kits were used to detect the content of inflammatory factors in serum: IL-1β, IL-18, IL-6, TNF-α, IL-1A, NF-ĸB, and the concentrations of IL-6, MDA, and SOD. The ELISA kits were purchased from Yisheng Biotechnology (Shanghai) Co., Ltd., and the determination steps were performed according to the instructions of the kit. Plasma biochemical indicators BUN, HDL-C, LDL-C, GLU, TP, ALB, TG, and TC were commissioned to Shanghai Liquid Quality Testing Company for testing.

[0087] ⑦ Microbial flora structure The 16S rDNA high-throughput sequencing technology was used to detect the microbial flora structure of the intestinal flora. The specific steps are as follows: Fresh colon contents were collected and frozen at -80°C for testing. In this experiment, the QIAamp DNA Stool MiniKit kit was used to extract total microbial DNA. The kit was purchased from QIAGEN, Germany, and the extraction steps were performed according to the kit instructions. The sample DNA concentration was determined using the NanoDrop ultra-micro spectrophotometer from Thermo Fisher Scientific. The samples were amplified and sequenced using the Illumina Miseq PE3000 high-throughput sequencing platform, and the amplification and sequencing primers were 27F and 1492R universal primers.

[0088] Among them, the nucleotide sequence of 27F is shown in SEQ ID NO. 2: 5’-AGAGTTTGATCCTGGCTCA-3’.

[0089] The nucleotide sequence of 1492R is shown in SEQ ID NO. 3: 5’-GGTTACCTTGTTACGACTT-3’.

[0090] 16S sequencing was completed by Beijing Ovisen Gene Technology Co., Ltd.

[0091] II. Results 2.1 Effects of Lactobacillus plantarum on the growth performance of weaned rabbits As can be seen from Table 6, compared with the control group, different combinations L. plantarum The QZF fermented feed had significant effects on the ADG of weaned rabbits from d1 to d21, d21 to d42, and d42 to d70 ( P <0.01), which could significantly improve the growth rate of weaned rabbits, indicating that adding lactic acid bacteria to the feed for fermentation was beneficial to improving the growth performance of weaned rabbits.

[0092] Table 6 Results of the effects of Lactobacillus plantarum on the growth performance of weaned rabbits Note: ADG represents average daily gain. A: Control group; B: Lactic acid bacteria group; C: Lactic acid bacteria + yeast group; D: Lactic acid bacteria + bacillus.

[0093] 2.2 Effects of Lactobacillus plantarum on the immune organ indexes of weaned rabbits As can be seen from Table 7, compared with the control group, different combinations L. plantarum The QZF fermented feed had no obvious effects on the spleen, heart, liver, lungs, and kidneys ( P (0.05).

[0094] Table 7 Results of the effects of Lactobacillus plantarum on the immune organ indexes of weaned rabbits Note: A: Control group; B: Lactic acid bacteria group; C: Lactic acid bacteria + yeast group; D: Lactic acid bacteria + bacillus.

[0095] 2.3 Effects of Lactobacillus plantarum on the plasma biochemical indexes of weaned rabbits As can be seen from Table 8, compared with the control group, different combinations L. plantarum The QZF fermented feed had significant changes in BUN and HDL-C in the plasma of weaned rabbits (P<0.05). Among them, L. plantarum The QZF single-fermented feed could increase the content of urea nitrogen BUN in the plasma of weaned rabbits, and BUN is a protein metabolite; L. plantarumThe combined fermentation of QZF and yeast in feed can increase the HDL-C content in the plasma of weaned rabbits. L. plantarum QZF fermented feed can improve the utilization rate of protein in feed by weaned rabbits and improve the cardiovascular function of animals. L. plantarum QZF fermented feed had no significant effect on GLU, TP, ALB, TG, TC, and LDL-C in weaned rabbits ( P >0.05).

[0096] Table 8 Effects of Lactobacillus plantarum on plasma biochemical parameters of weaned rabbits Note: A: control group, B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0097] 2.4 Effect of Lactobacillus plantarum on the antioxidant function of plasma in weaned rabbits It can be seen from Table 9 that compared with the control group, different combinations L. plantarum QZF fermented feed had a very significant effect on the CAT content in the plasma of weaned rabbits (P<0.01). 2 O 2 The content changed significantly (P<0.05).

[0098] Table 9 Effects of Lactobacillus plantarum on the antioxidant function of weaned rabbit plasma Note: A: control group, B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0099] 2.5 Effects of Lactobacillus plantarum on the anti-stress function of plasma in weaned rabbits It can be seen from Table 9 that compared with the control group, different combinations L. plantarum Feeding QZF fermented feed can significantly increase the levels of ALD (aldosterone) and CORT (cortisol) in the plasma of weaned rabbits. P <0.01). The test results show that different combinations L. plantarum Feeding QZF fermented feed can increase the levels of ALD and CORT in the plasma of weaned rabbits, thereby improving the animals' anti-stress ability, which is of great significance for alleviating weaning stress in young animals.

[0100] Table 9 Effects of Lactobacillus plantarum on the anti-stress function of plasma in weaned rabbits Note: A: control group, B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0101] 2.6 Effects of Lactobacillus plantarum on immunoglobulin function in weaned rabbits It can be seen from Table 10 that compared with the control group, different combinations L. plantarum QZF fermented feed has a very significant effect on the levels of IgA, IgM and IgG in the plasma of weaned rabbits ( P <0.01), can significantly increase the levels of IgA, IgM and IgG in the plasma of weaned rabbits. The experimental results show that L. plantarum QZF fermented feed can increase the content of immunoglobulin in the plasma of weaned pigs, thereby improving the disease resistance of animals.

[0102] Table 10 Effects of Lactobacillus plantarum on immunoglobulin function in weaned rabbits Note: A: control group, B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0103] 2.7 Effects of Lactobacillus plantarum on intestinal mucosal function in weaned rabbits It can be seen from Table 11 that compared with the control group, different combinations L. plantarum QZF fermented feed had a very significant effect on the levels of sIgA, IL-1α and IL-2 in the intestine of weaned rabbits ( P <0.01). Specifically, compared with the control group, L. plants QZF and yeast combined fermentation feed (Group C) and L. plantarum QZF and Bacillus co-fermented feed (Group D) can significantly increase the sIgA content in the small intestine and colon. sIgA is an antibody that plays a leading role in the intestinal mucosal immune system. It is the first line of defense against pathogens adhering to and colonizing the intestinal mucosa. The test results show that L. plants The combination of QZF and yeast or spores in fermented feed can enhance the intestinal mucosal immunity of animals, reduce intestinal inflammatory response, and improve intestinal barrier function. L. plantarum QZF fermented feed can increase the content of IL-1α and IL-2 in the small intestine of weaned rabbits. Both IL-1α and IL-2 are important immunomodulatory factors that can affect the proliferation, differentiation and function of immune cells and play a key role in intestinal inflammatory response. The experimental results show that different combinations of L. plantarum QZF fermented feed can enhance the immune response of the intestinal mucosa and help maintain the balance and stability of the intestinal immune system.

[0104] Table 11 Effects of Lactobacillus plantarum on intestinal mucosal function in weaned rabbits Note: A: Control group; B: Lactobacillus group; C: Lactobacillus + yeast group; D: Lactobacillus + bacillus.

[0105] 2.8 Effect of Lactobacillus plantarum on intestinal enzyme activity function of weaned rabbits As can be seen from Table 12, compared with the control group, different combinations L. plantarum The QZF fermented feed had a highly significant change in the amylase content in the duodenum of weaned rabbits ( P <0.01), and significantly increased the amylase content in the duodenum, indicating that different combinations L. plantarum The QZF fermented feed could improve the absorption rate of starch in the feed nutrition of weaned rabbits, improve the utilization rate of energy, and thus improve the feed utilization rate.

[0106] Table 12 Results of the effect of Lactobacillus plantarum on intestinal mucosal function of weaned rabbits Note: A: Control group; B: Lactobacillus group; C: Lactobacillus + yeast group; D: Lactobacillus + bacillus.

[0107] 2.9 Effect of Lactobacillus plantarum on intestinal morphology of weaned rabbits As can be seen from Table 13, Table 14, and Table 15, compared with the control group, different combinations L. plantarum The QZF fermented feed had significant changes in the morphology of the duodenum, jejunum, and ileum in weaned rabbits ( P <0.05). For the duodenum, L. plants The QZF fermented feed could increase the length of the villi and the number of epithelial cells in the duodenum, L. plants The QZF fermented feed in combination with yeast or bacillus could increase the villus height and crypt depth; for the jejunum, L. plants The QZF fermented feed in combination with bacillus could increase the number of epithelial cells in the villi, and different combinations L. plants The QZF fermented feed could significantly increase the number of epithelial cells in the villi per unit length; for the ileum, L. plants The QZF fermented feed could increase the thickness of the epithelial layer of the villi. Although different combinations L. plantarum The QZF fermented feed slightly reduced the number of epithelial cells in the villi, but did not affect the number of epithelial cells in the villi per unit length in the ileum. It shows that L. plants The QZF fermented feed had a certain improvement effect on the morphology of different small intestinal segments and might have a positive effect on improving the digestibility.

[0108] Table 13 One of the results of the effect of Lactobacillus plantarum on intestinal morphology of weaned rabbits Note: A: control group, B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0109] Table 14 Effect of Lactobacillus plantarum on intestinal morphology of weaned rabbits (result 2) Note: A: treatment group B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0110] Table 15 The effect of Lactobacillus plantarum on the intestinal morphology of weaned rabbits (result 3) Note: A: treatment group B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0111] 2.10 Effects of Lactobacillus plantarum on cecal parameters in weaned rabbits It can be seen from Table 16 that compared with the control group, different combinations L. plantarum QZF fermented feed had a very significant effect on the NH3-N content in the cecum of weaned rabbits (P<0.01). The increase in NH3-N content in the cecum may be related to the enhanced metabolic activity of intestinal microorganisms.

[0112] Table 16 Effects of Lactobacillus plantarum on cecal parameters of weaned rabbits Note: A: treatment group B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0113] 2.11 Effects of Lactobacillus plantarum on intestinal immune gene expression in weaned rabbits The gene expression in jejunum is shown in Figure 3 .

[0114] Depend on Figure 3 It can be seen that compared with the control group, different combinations L. plantarum QZF fermented feed had a very significant effect on the gene expression of IL-4 and IFN-r in weaned rabbits (P<0.01). The increase in IL-4 content in the jejunum usually indicates that there is inflammation or a strong immune response in the body, and the treatment group with probiotics can significantly inhibit the IL-4 content.

[0115] As can be seen from the above figure, the IFN-r content in the control group is higher than that in the treatment group, indicating that the treatment group with probiotics can significantly inhibit the IFN-r content.

[0116] The relative abundance of 16s microbial intestinal flora is shown in Table 17: Table 17 Relative abundance of 16s microbial intestinal flora Note: A: control group, B: lactic acid bacteria group; C: lactic acid bacteria + yeast group; D: lactic acid bacteria + spores.

[0117] Example 3: Research and development of composite microecological preparations and application of solid and liquid microbial fermentation feed technology 1. Test methods 1. Experimental Grouping According to Example 1, obtain L. plantarum QZF related fermentation performance data, to carry out the application research of combined microecological preparations for pregnant and lactating sows. 45 lactating sows with good health and similar gestational age were selected and randomly divided into 3 treatment groups, 15 in each treatment, and 1 replicate for each sow. The experimental period was from 90 days of gestation to weaning. The specific experimental design is shown in Table 18, where Bacillus subtilis was purchased from Xiamen Huiying Animal Technology Co., Ltd.

[0118] Table 18 Experimental design for pregnant and lactating sows Note: The "fermented water-Lactobacillus plantarum" in experimental group A is water fermented with Lactobacillus plantarum, and the "fermented water-mixed" in experimental group C is mixed fermentation of Lactobacillus plantarum and Bacillus subtilis. The fermented water formula production process is shown below.

[0119] Fermentation water formula and production process: Based on feed mass as the measurement basis, the specific composition is as follows: 100 portions of basic daily diet, 3 portions of seed liquid, 2 portions of glucose, 1000 portions of warm water at 30-38°C, sealed and fermented at room temperature of 25°C for 2 days.

[0120] Among them, pregnant and lactating sows come from the Shunchang breeding base of Fujian Huatian Agriculture and Animal Husbandry Technology Co., Ltd.

[0121] The basal diet met the nutritional requirements of the pigs during the experiment, as in Example 1.

[0122] The specific feeding management and safety production management details of all the sows in the above groups refer to the relevant system of Shunchang Breeding Base of Fujian Huatian Agriculture and Animal Husbandry Technology Co., Ltd. in Nanping City, Fujian Province. At the beginning of the experiment, the sows were evenly grouped according to weight and gestational age. Epidemic prevention was carried out according to the normal epidemic prevention process, and the pens were kept clean and the pig houses were well ventilated. At the beginning of the trial period, the corresponding feed was added according to the group feeding. The feed was weighed before feeding the group feed to calculate the feed intake. The free feeding method was adopted, and the feed was fed 3 times a day: 08: 00, 14:00, and 20:00. Free drinking water was adopted, and water shortage was strictly prohibited. The health status of each group and each pen was recorded daily, including constipation, death, etc.

[0123] 2. Measurement indicators and methods 2.1. Determination of sow reproductive performance Record the birth process, total litter size, live piglets, stillbirths, mummies, birth weight, birth litter weight, weaning age and weaning litter weight.

[0124] 2.2. Diarrhea rate in piglet farrowing room During the experimental period, at fixed times every day: starting at 9:00 am and 16:00 pm, check and record the fecal morphology scores of piglets as shown in Table 19. When the diarrhea score is ≥ 2, it is considered diarrhea. Diarrhea rate (%) = diarrhea pig days / (total number of pigs × experimental days)*100.

[0125] Table 19 Diarrhea score reference 2.3 Piglet growth performance On the 7th and 14th days after farrowing and on the day of weaning, the number of surviving piglets in each litter and the weight of each piglet were recorded. The piglet survival rate and average daily weight gain were calculated.

[0126] 2.4. Microbial flora structure Using 16S rDNA high-throughput sequencing technology, the feces of sows and piglets were used as materials to detect the microbial flora structure of the intestinal flora. Among them, 16S rDNA high-throughput sequencing was commissioned to Wuhan Sewell Biotechnology Co., Ltd.

[0127] 2. Results 1. Effect of fermented feed on sow reproductive performance Depend on Figure 4 As shown in Figures A and B, compared with the control group, the addition of Lactobacillus plantarum and the feed fermented with Lactobacillus plantarum and Bacillus subtilis can increase the total litter size and the number of live piglets born in the litter.

[0128] Depend on Figure 4 As shown in Figures C and D, compared with the control group, adding feed fermented with Lactobacillus plantarum can effectively reduce the number of stillbirths and mummies in a litter.

[0129] 2. Effect of fermented feed on growth performance of piglets Depend on Figure 5 It can be seen that compared with the control group, the addition of Lactobacillus plantarum and the feed fermented with Lactobacillus plantarum and Bacillus subtilis can improve the growth performance of piglets and increase their body weight at 7 days, 14 days and weaning. Among them, the weight gain effect of adding Lactobacillus plantarum fermentation to the feed is the most obvious on the 14th day.

[0130] 3. Effect of fermented feed on diarrhea in piglets Depend on Figure 6It can be seen that compared with the control group, Lactobacillus plantarum QZF fermented feed and L. plantarum The combination of QZF and Bacillus fermented feed can effectively reduce the diarrhea rate of piglets, especially the use of Lactobacillus plantarum fermented feed alone has the best effect on anti-piglet diarrhea.

[0131] In summary, feed fermented with Lactobacillus plantarum QZF can significantly improve the growth performance of weaned rabbits and improve intestinal health. When used in combination with yeast or Bacillus, it has better effects in different aspects. Feed fermented with Lactobacillus plantarum QZF can improve the reproductive performance of lactating sows, improve the growth performance of piglets, reduce the diarrhea rate of piglets, and enhance immunity. The experimental results of the two animals show that Lactobacillus plantarum fermentation L. plantarum QZF feed is beneficial to the growth and development and intestinal health of weaned animals. This strain has a good effect in improving intestinal health, relieving stress, enhancing disease resistance and improving growth performance.

[0132] It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. In order to avoid redundancy, the present invention describes a preferred embodiment.

[0133] Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they understand the basic inventive concepts, and all such changes and modifications fall within the scope of the present invention.

Claims

1. A strain of Lactobacillus plantarum QZF capable of improving intestinal health of animals, characterized in that: The Lactobacillus plantarum ( Lactobacillus plantarum )QZF was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on July 22, 2024, with the deposit number CGMCC No. 31389.

2. Use of Lactobacillus plantarum QZF according to claim 1 in regulating animal intestinal health.

3. The use according to claim 2, characterized in that: The Lactobacillus plantarum QZF is used for promoting animal feeding.

4. A culture comprising the Lactobacillus plantarum QZF capable of improving the intestinal health of animals as claimed in claim 1.

5. A bacterial agent for improving the intestinal health of animals, characterized in that: The bacterial agent comprises the culture according to claim 4.

6. The bacterial agent according to claim 5, characterized in that The culture is a solid fermentation culture or a liquid fermentation culture.

7. The bacterial agent according to claim 5, characterized in that The culture includes a bacterial liquid of Lactobacillus plantarum QZF; The bacterial liquid is a fermentation liquid obtained by inoculating the plant lactobacillus QZF into a separation and purification medium; The separation and purification medium includes any one of LB solid medium, LB liquid medium and MRS broth medium.

8. The bacterial agent according to claim 5, characterized in that The bacterial agent is processed into an agriculturally acceptable preparation; The dosage form of the bacterial agent is any one of liquid, powder and granule.

9. Use of the culture according to claim 4 or the bacterial agent according to claim 5 in preparing a product for improving the intestinal health of animals, characterized in that: The product comprises any one or more of fermented feed, preservatives, silage additives and livestock and poultry intestinal flora stabilizers.

10. A product for improving the intestinal health of animals, characterized in that: The method comprises the culture according to claim 4 or the bacterial agent according to claim 5, and an auxiliary material acceptable to food and / or health care products.

Images