Application of lactobacillus fermentum to improvement of fatty liver hemorrhagic syndrome of laying hens in delayed feeding period

Abstract

The invention belongs to the technical field of microorganisms, and relates to application of lactobacillus fermentum to improvement of a fatty liver hemorrhagic syndrome of laying hens in a delayed rearing period, in particular to application of lactobacillus fermentum to preparation of a product for improving the fatty liver hemorrhagic syndrome of the laying hens in the delayed rearing period. Lactobacillus fermentum is prepared into a zymophyte solution, and the zymophyte solution is administered to the laying hens with the fatty liver in an intragastric administration manner, so that the lactobacillus fermentum can effectively treat the hemorrhagic syndrome of the laying hens. Experiments prove that the lactobacillus fermentum can reduce the liver lipid and blood fat levels of laying hens, increase the antioxidant activity, reduce inflammation and relieve the fatty liver hemorrhagic syndrome of the laying hens, so that the death and culling rate of the laying hens is reduced, a new product is provided for treatment intervention of the fatty liver hemorrhagic syndrome of the laying hens, and a new application of the lactobacillus fermentum is provided.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to the application of fermented lactobacillus in improving fatty liver hemorrhagic syndrome in extended-rearing laying hens, and particularly relates to the application of fermented lactobacillus in preparing a product for improving fatty liver hemorrhagic syndrome in extended-rearing laying hens. Background Art

[0002] Extended rearing of laying hens is particularly important. High-yielding laying hens lay more than 500 eggs at 700 days of age. However, in the late egg-laying period, due to the aging of various organs, laying hens are often accompanied by a decline in production performance and egg quality, such as a decline in egg production rate, eggshell strength and Haugh units, which brings huge losses to the poultry farming and egg processing industries and is a bottleneck problem in the extended rearing plan for laying hens. At the same time, in recent years, with the continuous increase in the scale of laying hen farming, the stocking density of laying hen cages has increased accordingly. Problems of overnutrition and unbalanced feed supply often occur in laying hen production, causing metabolic disorders in laying hens, especially lipid metabolism disorders, leading to increased blood lipid content in laying hens, causing a large amount of lipid substances to be deposited in adipose tissue, causing fatty liver hemorrhage syndrome in laying hens, namely fatty liver hemorrhage syndrome, FLHS, which is a common nutritional metabolic disease characterized by a large amount of fat deposition and bleeding in the liver, which can lead to a variety of adverse effects in the short term, such as a decrease in egg production rate of the flock, delayed and shortened peak egg production, etc., and often occurs in the peak egg production period and late egg production period of commercial caged laying hens, accounting for 40%~70% of the hen mortality rate, seriously threatening the economic benefits of the laying hen farming industry.

[0003] Lactobacillus fermentum, Lactobacillus fermentum As a kind of symbiotic bacteria, a type of lactic acid bacteria, it can colonize in the intestine, produce acid, has adhesion and tolerance, and has many benefits such as regulating intestinal microbial balance, enhancing biological barriers, participating in immune system regulation, lowering cholesterol and blood pressure. It is an important type of beneficial intestinal bacteria. It is widely known for its antibacterial activity, cholesterol degradation ability, immune activity, antioxidant properties, relief of liver and intestinal damage and other probiotic properties. Studies have reported that fermented lactobacillus can be widely used in the treatment of various diseases, but it has not been reported in alleviating fatty liver hemorrhagic syndrome in laying hens, improving production performance of laying hens and egg quality.

[0004] Current treatments and shortcomings for fatty liver in laying hens: Due to the lack of a clear in vivo diagnostic method for fatty liver syndrome in laying hens, the disease process is slow, and there are differences in the disease stages among the diseased individuals in the flock, which makes it very difficult to prevent and treat fatty liver in laying hens in actual breeding. In traditional Chinese medicine, Chinese medicinal ingredients such as Ligustrum lucidum extract are often added to the diet of laying hens, but the success rate of prevention and treatment is low; Western medicine uses drugs and antioxidants for prevention and treatment, which has the potential safety hazard of polluting the environment. Therefore, under the background of the comprehensive ban on antibiotics in the feed industry, it is necessary to explore new responses and solutions for fatty liver in laying hens. Summary of the invention

[0005] In order to solve the defect of low prevention and treatment success rate in the prior art traditional Chinese medicine treatment methods, the present invention provides the use of fermented lactobacillus to improve fatty liver hemorrhagic syndrome in extended-rearing laying hens.

[0006] The invention provides application of fermented lactobacillus in preparing a product for improving fatty liver hemorrhagic syndrome in extended-rearing laying hens.

[0007] In some technical solutions of the present invention, the product is a laying hen feed additive or a prebiotic.

[0008] In some technical solutions of the present invention, the product is a liquid preparation or a solid preparation.

[0009] In some technical solutions of the present invention, when the product is a liquid preparation, the viable count of the fermented lactobacillus is 0.8×10 9 CFU / mL~1.2×10 9 CFU / mL; when the product is a solid preparation, the viable count of the fermented lactobacillus is 0.8×10 9 CFU / g~1.2×10 9 CFU / g.

[0010] In some technical schemes of the present invention, the preparation method of the liquid preparation comprises: inoculating the fermented lactobacillus into an MRS culture medium, activating it at 35° C. to 37° C. for 30 h to 40 h, to obtain the liquid preparation.

[0011] In some technical solutions of the present invention, the fermented Lactobacillus is inoculated into the MRS culture medium at 1% to 5% of the volume of the MRS culture medium.

[0012] In some technical solutions of the present invention, the number of activations is 1 to 3 times.

[0013] In some technical solutions of the present invention, the product is suitable for the entire growth cycle of the laying hen.

[0014] Preferably, the product is suitable for the egg-laying period of the laying hens.

[0015] More preferably, the laying hens are extended-care laying hens. The frequency of fatty liver in extended-care laying hens is relatively high. In order to increase the extended-care period of laying hens, this group is more representative. If there is a good improvement effect in the selected extended-care laying hens, it means that early dietary input can better improve the production performance of laying hens, protect the health of laying hens, and achieve the goal of producing 500 eggs in 100 weeks.

[0016] In some technical solutions of the present invention, the product is used to alleviate fatty degeneration of the liver of poultry.

[0017] In some technical solutions of the present invention, the product is used to reduce the blood lipid level of poultry.

[0018] In some technical solutions of the present invention, the product is used to reduce liver damage in poultry.

[0019] In some technical solutions of the present invention, the product is used to improve the production performance of poultry.

[0020] In some technical solutions of the present invention, the product is used to improve the quality of eggs produced by laying hens.

[0021] In some technical solutions of the present invention, the poultry includes laying hens.

[0022] Compared with the prior art, the present invention has the following beneficial effects: in the present invention, fermented lactobacillus is prepared into a fermented bacterial liquid, which is administered to laying hens with fatty liver by gavage, and it is found that the fermented lactobacillus can effectively reduce the liver fat and blood lipid levels of laying hens, increase antioxidant activity, reduce inflammation and alleviate fatty liver hemorrhagic syndrome of laying hens, thereby reducing the mortality rate of laying hens, providing a new product for the treatment intervention of fatty liver hemorrhagic syndrome of laying hens, and providing a new use of fermented lactobacillus.

[0023] The preparation method of the fermented bacterial liquid provided by the present invention is simple, and the fermented lactic acid bacteria are easy to obtain. Therefore, the cost of using the fermented lactic acid bacteria to treat fatty liver hemorrhagic syndrome in laying hens is low.

[0024] The fermented lactobacillus provided by the present invention can also avoid the problem of environmental pollution caused by western medicine, and provides a new approach and solution for improving fatty liver in laying hens. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The morphological and histological observation results of the liver of laying hens in different treatment groups are shown in the figure; Figure 1 A represents the morphological observation of laying hen liver in different treatment groups; Figure 1 B shows the results of hematoxylin-eosin staining and oil red O staining of laying hen livers in different treatment groups. Figure 1The first row of figures in B represents the staining results at a scale of 50 μm, the second row of figures represents the staining results at a scale of 20 μm; and the third row of figures represents the staining results at a scale of 20 μm.

[0026] Figure 2 This is a graph showing the results of liver index determination of laying hens in different treatment groups.

[0027] Figure 3 The results of blood lipid determination of laying hens in different treatment groups are shown in the figure; Figure 3 A in the figure represents the determination results of triglycerides in different treatment groups; Figure 3 B in the figure represents the result of total cholesterol determination; Figure 3 The C in the figure represents the measurement result of high-density lipoprotein cholesterol; Figure 3 D in the figure represents the measurement result of low-density lipoprotein cholesterol; Figure 3 E in FIG. 1 represents the measurement result of very low density lipoprotein cholesterol.

[0028] Figure 4 The results are the results of the determination of the antioxidant capacity of the liver of laying hens in different treatment groups; Figure 4 A in the figure represents the result of the determination of catalase; Figure 4 B in the figure shows the measurement result of total superoxide dismutase.

[0029] Figure 5 The results of the determination of serum antioxidant capacity of laying hens in different treatment groups are shown in Table 1. Figure 5 A in the figure represents the result of the determination of catalase; Figure 5 B in the figure shows the measurement result of total superoxide dismutase.

[0030] Figure 6 The results of liver enzyme activity determination in the liver of laying hens in different treatment groups are shown in the figure; Figure 6 A in the figure represents the result of the measurement of alanine aminotransferase in the liver; Figure 6 B in the figure shows the result of the measurement of aspartate aminotransferase in the liver; Figure 6 C in FIG. 1 shows the results of the measurement of alkaline phosphatase in the liver.

[0031] Figure 7 The results of the determination of liver enzyme activity in the serum of laying hens in different treatment groups are shown in Figure 2. Figure 7 A in the figure represents the result of the determination of alanine aminotransferase in serum; Figure 7 B in the figure shows the result of measuring aspartate aminotransferase in serum; Figure 7 C in FIG. 1 is a graph showing the measurement results of alkaline phosphatase in serum. DETAILED DESCRIPTION

[0032] The present invention is described in detail below in conjunction with specific examples, but it should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0033] Current treatments and shortcomings for fatty liver in laying hens: Due to the lack of a clear in vivo diagnostic method for fatty liver syndrome in laying hens, the disease process is slow, and there are differences in the disease stages among the sick individuals in the flock, which makes it very difficult to prevent and treat fatty liver in laying hens in actual breeding. In traditional Chinese medicine, Chinese medicinal ingredients such as Ligustrum lucidum extract are often added to the diet of laying hens, but there are problems with low prevention and treatment success rates and high costs of prevention and treatment plans; Western medicine uses drugs and antioxidants for prevention and treatment, which also have high costs and potential safety hazards of polluting the environment. Therefore, under the background of comprehensive antibiotic ban in the feed industry, it is necessary to explore new responses and solutions for fatty liver in laying hens.

[0034] The present invention provides the use of fermented lactobacillus to improve fatty liver hemorrhagic syndrome in extended-rearing laying hens, and specifically, provides the use of fermented lactobacillus in the preparation of a product for improving fatty liver hemorrhagic syndrome in laying hens. Specifically, in the present invention, fermented lactobacillus is prepared into a fermented bacterial liquid, and administered to fatty liver laying hens by gavage, and it is found that fermented lactobacillus can effectively treat hemorrhagic syndrome in laying hens. The fermented lactobacillus provided by the present invention can effectively reduce the liver fat and blood lipid levels of laying hens, increase antioxidant activity, reduce inflammation and alleviate fatty liver hemorrhagic syndrome in laying hens, thereby reducing the mortality rate of laying hens, providing a new product for the treatment intervention of fatty liver hemorrhagic syndrome in laying hens, and providing a new use of fermented lactobacillus.

[0035] Method for constructing a fatty liver model in laying hens: Using a high-energy, low-protein feed to induce a fatty liver pathological model in laying hens is a classic method for establishing a model, and is also the natural pathological form of fatty liver in laying hens, which is consistent with actual production conditions. The energy content exceeding the normal value and the lower protein level make it difficult for lipids to combine with a sufficient amount of protein to form lipoproteins. As the fat transport rate in the laying hen liver decreases, the fat in the laying hen liver continues to accumulate, ultimately achieving the purpose of inducing fatty liver in laying hens.

[0036] The Jingfen No. 6 laying hens used in the present invention were purchased from Beijing Huadu Yukou Poultry Co., Ltd.

[0037] The fermented lactobacillus used in the present invention Lactobacillus fermentum The strain number is BNCC255038, which was purchased from Beina Biotechnology.

[0038] The abbreviations used in the following examples have the following respective definitions: TG, triglyceride; TC, total cholesterol; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; VLDL-C, very low-density lipoprotein cholesterol; CAT, catalase; T-SOD, total superoxide dismutase; ALT, alanine aminotransferase; AST, aspartate aminotransferase; ALP, alkaline phosphatase.

[0039] Embodiment 1: 1. Animal testing The laying hen model selected in this embodiment is a fatty liver model induced by a high-energy, low-protein diet. The laying hens of Jingfen No. 6 were continuously fed a high-energy, low-protein diet for 10 weeks, during which fermented lactobacillus was intervened. The specific details are as follows: Before the start of this experiment, the laying hen house was cleaned, the feeding trough was cleaned, the ventilation was disinfected, and the chicken cages were grouped and numbered. 80 healthy laying hens of Jingfen No. 6, weighing close to 1.96±0.15kg, aged 79 weeks, i.e. 560 days, were selected and randomly divided into 4 treatment groups, with 20 chickens in each treatment group. All chickens were pre-fed for 1 week and uniformly fed a basic diet, two meals a day, 120g / day per chicken. During this period, the laying hens were replaced to ensure that the initial weight and egg production rate of the laying hens in each treatment group were basically the same. The feeding period is 80 to 90 weeks. The experimental design is shown in Table 1. The CON group was fed a basic diet, the HFLP group was fed a high-energy, low-protein diet, and the HFLP + Limosilactobacillus fermentum The LF group was fed with a high-energy, low-protein diet and gavaged with 1×10 9 CFU / mL of 1 mL of fermented lactobacillus liquid, the HFLP+HLF group was fed a high-energy, low-protein diet, and the intragastric concentration was 1×10 9 CFU / mL in 1mL of heat-killed fermented lactobacillus solution.

[0040] The experiment adopted a three-layer fully stepped arrangement of metal cages, with single cages and single cages, and the specifications of the metal cages were 40×35×40cm. The daily feeding and management of laying hens during the pre-feeding period and the experimental period were carried out in accordance with the "Jinghong Commercial Layer Feeding and Management Manual" of Yukou Laying Hens: feeding at 08:30 and 14:30 every day, the chickens were free to eat, and the chickens were free to drink water with nipple drinkers. The chicken house uses a fully automatic temperature sensing system to control the ventilation and wet curtain operation in the house. The temperature of the chicken house is maintained at 26℃±2℃, and the humidity is maintained at 50%±10%. During the experiment, the chicken house implemented a system of 16h light and 8h dark treatment, and the light intensity was maintained at 10Lux~20Lux.

[0041] Preparation method of fermented lactobacillus liquid: take out the fermented lactobacillus liquid preserved in glycerol from −80°C, inoculate it into MRS culture medium at a volume ratio of 2%, activate it twice at 37°C, each time for 36 hours; put the activated liquid into 50mL sterile enzyme-free centrifuge tubes for later use.

[0042] By selecting any value within the parameter range in the above method, a fermented lactobacillus culture solution can be obtained. Specifically, the following method is used in this embodiment: the fermented lactobacillus culture solution preserved in glycerol is taken out from -80°C, and is inoculated into MRS culture medium at a volume ratio of 2%, and activated twice at 37°C, each time for 36 hours; the activated culture solution is placed in a 50mL sterile enzyme-free centrifuge tube for standby use.

[0043] Preparation of heat-killed bacterial solution: Take out the above bacterial solution, boil it in boiling water for 15 minutes to prepare heat-killed bacterial solution, gavage it once every two days, 1 mL of bacterial solution each time, prepare fresh bacterial solution in advance every week and store it in a 4°C refrigerator.

[0044] The implementation location of this embodiment is the poultry experimental base of China Agricultural University in Zhuozhou, Hebei. The experimental design and feeding management comply with the regulations of the Animal Care and Use Committee of China Agricultural University on the management of experimental animals.

[0045] Table 1 Design of the fermentation lactobacillus experiment

[0046] 2. Experimental diet formulation, nutritional level and design The experimental diet formula was designed according to the "Chicken Feeding Standard" NY / T33-2004. The specific information on feed ingredients and nutritional value is shown in Tables 2 and 3.

[0047] Table 2 Composition and nutritional level of experimental diets

[0048] Note: “-” means there is no such item.

[0049] Table 3 Nutrient levels of experimental diets

[0050] (1) The trace element premix provides 6.8 mg copper, 66 mg iron, 83 mg zinc, 80 mg manganese and 1 mg iodine per kilogram of diet.

[0051] (2) Multivitamin premix provides per kilogram of diet: vitamin A 11700IU, vitamin D3 3600IU, vitamin E 21IU, vitamin K3 4.2mg, thiamine 3mg, riboflavin 10.2mg, folic acid 0.9mg, calcium pantothenate 15mg, niacin 45mg, pyridoxine 5.4mg, vitamin B12 24ug, biotin 150µg.

[0052] 4. Sample Collection and Preparation One day before the end of the experiment, all laying hens were deprived of feed for 8 hours, the sampling room was disinfected with ultraviolet light in advance, the sampling table and clean bench were disinfected with alcohol, and the sampling tools such as surgical scissors, scalpels, and tweezers were sterilized at 121°C for 30 minutes using a vertical steam pressure sterilizer model BKQ-B50II. After the formal trial period, the venous blood under the wings of the experimental chickens was collected in a procoagulant blood collection tube, placed in a 4°C refrigerator for about 4 hours, and centrifuged at 4°C for 15 minutes at 1500r / min. The separated serum was stored in a −80°C refrigerator for the detection of serum biochemical indicators.

[0053] At the end of the experiment, laying hens were killed, and the livers of all laying hens were taken and photographed. The livers of laying hens were collected, weighed, and the relative weight was calculated, which was the weight of tissue and organ g / body weight kg. Part of the liver tissue was quickly placed in liquid nitrogen and stored in a −80°C refrigerator; at the same time, the left lobe of the liver of appropriate size was placed in a 4% paraformaldehyde fixative for fixation, and then the liver tissue blocks fixed in the 4% paraformaldehyde solution were cut into appropriate sizes, routinely embedded in paraffin, stained with hematoxylin-eosin, i.e. H&E staining and oil red O staining, and observed under an optical microscope. At the same time, inflammatory indicators, antioxidant indicators, and lipid levels in the liver and serum were detected.

[0054] The tissue samples of the jejunum and cecum in the middle section were collected. After the intestinal contents were rinsed with PBS without mechanical damage, they were cut into appropriate lengths and placed in 4% paraformaldehyde fixative for fixation. The liver tissue blocks fixed in 4% paraformaldehyde solution were then cut into appropriate sizes for routine paraffin embedding, H&E staining and Alcian blue staining, and optical microscope observation. At the same time, the remaining jejunum was cut longitudinally, and the intestinal mucosa was scraped off with a glass slide and placed in sterile cryopreservation tubes for intestinal flora analysis; the remaining cecal contents were taken and placed in sterile cryopreservation tubes for intestinal flora analysis. Except for the samples used for analysis, the remaining tissue samples were stored at −80°C until the next analysis.

[0055] 5. Test results 1) Effects of fermented lactobacillus on production performance of laying hens during extended-life period As can be seen from Table 4, compared with the CON group, the HFLP group caused a significant decrease in the daily egg production of laying hens and a significant increase in the feed-egg ratio; while compared with the HFLP group, the egg production rate and daily egg production of laying hens in the HFLP+LF group were significantly increased, the feed-egg ratio was significantly reduced, and the treatment effect was better than that of the heat-killed treatment group, indicating that fermented lactobacillus can significantly affect the production performance of laying hens in the extended rearing period.

[0056] Table 4 Effects of fermented lactobacillus on production performance of laying hens during extended period

[0057] Note: Different letters represent significant differences. P <0.05.

[0058] 2) Effect of fermented lactobacillus on the quality of laying eggs during extended culture period After the test, all eggs from each treatment were collected for egg quality measurement. First, the eggshell strength of the sample eggs was measured and recorded using the eggshell strength tester model EFR.01. Then, the eggs were gently broken and the egg white and yolk were placed intact on the detection area of ​​the multifunctional egg quality tester model DET-6500. After pressing the start button and waiting for a few seconds, the egg albumen height, yolk color, and Haugh unit of the egg were automatically displayed on the multifunctional egg quality tester. The test results are valid only if the egg white and yolk are intact. Otherwise, the results are invalid. After one test, rinse the test tray and wipe it dry, and then repeat the above process. The eggshell thickness was measured using a spiral micrometer. A small portion of the eggshell was taken from the top, equator, and blunt end, and the eggshell membrane was peeled off to measure the eggshell thickness. The measurement was performed twice and recorded. Finally, the valid results of the above tests were recorded in the experimental record book.

[0059] As can be seen from Table 5, compared with the HFLP group, the HFLP+LF group significantly increased the protein level of laying hens' eggs, and there were no significant differences in other indicators, indicating that adding fermented lactobacillus can improve the egg quality of laying hens.

[0060] Table 5 Effect of fermented lactobacillus on the quality of laying eggs during extended culture period

[0061] 3) Fermented Lactobacillus can alleviate HFLP-induced liver steatosis in FLHS laying hens After the experiment, the livers of laying hens in different groups were taken for morphological and histological observation. Figure 1As shown in A and B, the liver of the CON group had a smooth surface, normal color and no bleeding spots, while the liver of the HFLP group was yellowish in color, with a large number of bleeding spots and blood spots, and was soft and fragile; the bleeding spots in the liver of the HFLP+LF group were significantly reduced, and the color returned to normal, while the liver of the HFLP+HLF group was still slightly yellow in color, with bleeding spots and blood spots, and was soft and fragile. H&E staining of the paraffin sections of the liver showed that the liver cells of the CON group had a clear and neatly arranged structure, with clear cell boundaries, and no obvious fat vacuoles; the liver cells of the HFLP group had fuzzy boundaries and disordered arrangement, with a large number of obvious fat vacuoles of varying sizes and numbers; the liver cells of the HFLP+LF group were obviously neatly arranged, and there were no obvious fat vacuoles; the liver cells of the HFLP+HLF group still had obvious fat vacuoles. From the oil red O staining of the frozen liver sections, it can be seen that there are very few red lipid droplets in the liver cells of the CON group; the number of lipid droplets in the HFLP group almost fills the entire field of view, all of which are red lipid droplets, and almost no normal liver cells can be seen; the number of lipid droplets in the HFLP+LF group is significantly reduced, and there are still obvious lipid droplets in the HFLP+HLF group. Figure 2 It can be seen that compared with the HFLP group, the HFLP+LF group significantly reduced the liver index, indicating that fermented lactobacillus can significantly improve fatty degeneration of the liver of laying hens.

[0062] 4) Fermented Lactobacillus can reduce blood lipid levels in laying hens with HFLP-induced FLHS Method for determination of lipids in serum: serum samples frozen at −20°C were thawed at room temperature, and the lipid metabolism indicators TG, TC, HDL-C, LDL-C, and VLDL-C in serum were determined according to the kit instructions of Nanjing Jiancheng Bioengineering Institute.

[0063] Serum lipid test results: Figure 3 As shown in AE, compared with the CON group, the HFLP group had increased TG, TC, HDL-C, LDL-C and VLDL-C levels; compared with the HELP group, the HFLP+LF group had significantly decreased TG and VLDL-C levels. P <0.05, and there was no significant difference in other parameters, but all showed a downward trend, indicating that fermented lactobacillus can regulate lipid metabolism disorders caused by FLHS in laying hens and improve liver function damage caused by FLHS.

[0064] 5) Fermented Lactobacillus can reduce the antioxidant capacity of HFLP-induced FLHS laying hens Method for determining liver antioxidant capacity: a) Thaw liver samples frozen at −80°C at room temperature and weigh 0.2 g into a 2 mL centrifuge tube.

[0065] b) Add 1.8 mL of normal saline and homogenize on ice.

[0066] c) Centrifuge at 3000 rpm, 4°C for 15 min.

[0067] d) Take 1 mL of supernatant for testing.

[0068] e) Use the Biotech BCA protein concentration assay kit to determine the protein content in the test solution.

[0069] f) The CAT and T-SOD activities of the test solution were determined using the kit produced by Nanjing Jiancheng Company.

[0070] The results of liver antioxidant capacity test: Figure 4 As shown in A and B, compared with the CON group, the HFLP group significantly decreased the content of T-SOD in the liver. P <0.05; compared with the HFLP group, the HFLP+LF group significantly increased the levels of CAT and T-SOD in the liver, P <0.05; compared with the HFLP+HLF group, the HFLP+LF group significantly increased the T-SOD content in the liver. The above results indicate that the addition of fermented lactobacillus significantly increased the reduction in liver antioxidant function caused by the high-energy, low-protein diet, and the effect was better than that of the heat-inactivated treatment group.

[0071] Method for determining serum antioxidant capacity: thaw the -20°C frozen serum samples at room temperature, and use the kit produced by Nanjing Jiancheng Company to determine the activities of CAT and T-SOD in the serum.

[0072] The results of serum antioxidant capacity test: Figure 5 As shown in A and B in Figure 3, compared with the CON group, the HFLP group significantly reduced the levels of serum CAT and T-SOD. P <0.05; compared with the HFLP group, HFLP+LF significantly increased the levels of CAT and T-SOD in serum, P <0.05; compared with the HFLP+HLF group, the HFLP+LF group significantly increased the levels of serum CAT and T-SOD, P <0.05 content. The above results show that the addition of fermented lactobacillus significantly reversed the decrease in serum antioxidant function caused by a high-energy, low-protein diet, and the effect was better than that of the heat-inactivated treatment group.

[0073] 6) Fermented Lactobacillus can reduce liver damage induced by HFLP in FLHS laying hens Method for determination of liver enzyme activity in liver and serum: liver samples frozen at −80°C and serum samples frozen at −20°C were thawed at room temperature, and the levels of ALT, AST, and ALP in liver and serum samples were determined using the kit produced by Nanjing Jiancheng Company.

[0074] Results of liver enzyme activity test in the liver: Figure 6 As shown in Figures AC, compared with the CON group, the HFLP group significantly increased the activities of AST and ALP in the liver. P <0.05; compared with the HFLP group, the HFLP+LF and HFLP+HLF groups significantly decreased the activities of ALT and ALP in the liver, P <0.05, but the relief effect of HFLP+LF group was better than that of HFLP+HLF group.

[0075] Results of serum liver enzyme activity test: Figure 7 As shown in Figures AC, compared with the CON group, the HFLP group had significantly increased serum AST and ALP activities. P <0.05; compared with the HFLP group, HFLP+LF significantly reduced the activities of ALT, AST and ALP in serum, P <0.05.

[0076] The above results indicate that fermented lactobacillus can reduce the increase in transaminase activity caused by FLHS in laying hens and inhibit the liver damage caused by FLHS in laying hens.

[0077] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0078] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Application of fermented lactobacillus in the preparation of products to improve fatty liver hemorrhagic syndrome in extended-rearing laying hens.

2. The use according to claim 1, characterized in that The product is a laying hen feed additive or a prebiotic.

3. The use according to claim 2, characterized in that The product is a liquid preparation or a solid preparation.

4. The use according to claim 3, characterized in that When the product is a liquid preparation, the viable count of the fermented lactobacillus is 0.8×10 9 CFU / mL~1.2×10 9 CFU / mL; when the product is a solid preparation, the viable count of the fermented lactobacillus is 0.8×10 9 CFU / g~1.2×10 9 CFU / g.

5. The use according to claim 4, characterized in that The preparation method of the liquid preparation comprises: inoculating the fermented lactobacillus into an MRS culture medium, activating the culture medium at 35° C. to 37° C. for 30 h to 40 h, and obtaining the liquid preparation.

6. The use according to claim 5, characterized in that The fermented lactobacillus is inoculated into the MRS culture medium according to 1% to 5% of the volume of the MRS culture medium.

7. The use according to claim 6, characterized in that The activation is performed once to three times.

8. The use according to claim 1, characterized in that The product is used for improving the production performance of laying hens.

9. The use according to claim 1, characterized in that The product is used for improving the quality of eggs produced by laying hens.

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