Application of moringa oleifera leaves in preparation of product for treating and / or relieving poultry fatty liver in later egg laying period

Abstract

The invention relates to the technical field of livestock breeding, in particular to application of horseradish tree leaves in preparation of a product for treating and / or relieving poultry fatty liver in the later egg laying period. The application research finds that the moringa oleifera leaves can reduce the fatty liver and blood fat level of the hyline brown laying hens in the later egg laying period, increase the antioxidant activity, reduce the death and culling rate and slow down the lowering trend of the egg laying rate.

Description

Technical Field

[0001] This invention relates to the field of animal husbandry technology, and in particular to the application of Moringa leaves in the preparation of products for treating and / or alleviating fatty liver in poultry during the late laying period. Background Technology

[0002] The egg-laying cycle of laying hens can be divided into the pre-laying period, peak laying period, and post-laying period. As laying hens enter the post-laying period, egg production begins to decline, and egg quality also decreases. This decline in egg production during the post-laying period is not only related to nutrient intake but also to the increasing number of free radicals in the body. Excessive free radicals can damage cells and affect organ function. This decline in production performance and egg quality directly impacts the economic benefits for farmers. In the current egg-laying hen market, extending the rearing period can significantly increase profitability. This strategy not only reduces the costs and labor associated with frequent brooding but also increases total egg production by extending the laying period, thereby maximizing profits.

[0003] However, as laying hens age in the later stages of egg production, their egg production performance and quality often decline, leading to losses for the poultry and egg processing industries. Furthermore, late-laying hens face liver lipid metabolism disorders, which can result in diseases such as fatty liver hemorrhage syndrome (FLHS), posing a potential threat to hen health and the economic benefits of poultry farming. FLHS is a common and serious metabolic disease in laying hen farming. With the expansion of laying hen farming scale and the increase in intensification, the incidence of FLHS is on the rise. Affected hens often exhibit symptoms such as decreased egg production performance, hepatic steatosis, and hemorrhage; in severe cases, it can lead to death, causing huge economic losses to the laying hen industry. Currently, there are limited prevention and control methods for FLHS, making the development of safe and effective prevention and control methods an urgent priority.

[0004] Moringa leaves are rich in various bioactive components, such as flavonoids, phenols, and polysaccharides, possessing multiple biological functions including antioxidant, anti-inflammatory, and lipid-regulating effects. Moringa leaves are a natural plant with excellent safety and rich functional content, listed as a feed ingredient, and have been proven to regulate lipid metabolism disorders in laying hens. Existing studies have shown that Moringa leaves and their extracts have potential in improving the metabolism of laying hens and reducing oxidative stress; however, research on their application is still in its early stages, especially lacking systematic exploration at the level of mechanisms regulating hepatic lipid metabolism. Whether it can serve as a potential drug for alleviating fatty liver in Hy-Line Brown laying hens during the later stages of egg production, and its specific mechanism of action, remains unclear. Exploring new solutions and approaches to fatty liver in laying hens during the later stages of egg production is of great significance. Summary of the Invention

[0005] This invention provides the application of Moringa leaves in the preparation of products for treating and / or alleviating fatty liver in poultry during the late laying period. Moringa leaves can reduce fatty liver and blood lipid levels in Hy-Line Brown laying hens during the late laying period, increase antioxidant activity, reduce mortality, and slow down the decline in egg production, thus solving the problems existing in the prior art.

[0006] The technical solution adopted in this invention is:

[0007] The use of Moringa leaves in the preparation of products for treating and / or alleviating fatty liver in late-laying poultry, namely late-laying Hy-Line Brown chickens, is provided by improving hepatic lipid droplet accumulation and reducing liver damage.

[0008] Furthermore, the fatty liver is fatty liver hemorrhage syndrome; the dosage of Moringa leaves as an additive is 5-10% of the feed weight.

[0009] Furthermore, the dosage of moringa leaves added is 5%.

[0010] Furthermore, the moringa leaves alleviate fatty liver in laying hens by improving the accumulation of lipid droplets in the liver and reducing liver damage in Hy-Line Brown laying hens during the later stages of egg production.

[0011] Furthermore, the main active ingredients in Moringa leaves include isoquercitrin, quercetin-3-O-rhamnoside, rutin, kaempferol-3-O-rutin, etc.

[0012] Furthermore, the above applications can improve and achieve any of the following functions in the later stages of egg production in laying hens:

[0013] S1) Reduce the accumulation of abdominal fat in poultry and / or reduce the abdominal fat weight in poultry;

[0014] S2) Relieves liver lesions in poultry;

[0015] S3) Maintain the integrity of the structure and morphology of poultry liver and / or alleviate steatosis in poultry liver;

[0016] S4) Reduce liver lipid levels in poultry;

[0017] S5) Reduces cholesterol and / or triglyceride levels in poultry serum;

[0018] S6) Reduces liver enzyme levels in poultry serum, alleviating liver damage;

[0019] S7) alleviates the sharp drop in egg production rate in poultry during the later stages of egg production.

[0020] The beneficial effects of this invention are:

[0021] This invention demonstrates that Moringa leaves can reduce fat deposition in the liver and abdomen of Hy-Line Brown laying hens during the late laying period, alleviate fatty liver hemorrhage syndrome, and thus reduce mortality rates, thereby improving the economic benefits of egg-laying hen farming. This study indicates that Moringa leaves, when used in optimized amounts, can serve as a novel and effective feed additive for alleviating liver lesions in poultry during the late laying period or for preparing products that alleviate liver lesions in poultry during this time, showing great promise for application in the egg-laying hen farming sector. Attached Figure Description

[0022] Figure 1 Images of liver sections, HE and Oil Red O stained sections, from Hy-Line Brown laying hens fed a basal diet during the late laying period;

[0023] Figure 2 Images of liver sections, HE and Oil Red O stained sections, from Hy-Line Brown laying hens fed a high-fat diet during late-laying periods;

[0024] Figure 3 The effect of Moringa leaves on serum biochemical parameters of Hy-Line Brown laying hens fed a high-fat diet during the late laying period;

[0025] Figure 4 The effect of Moringa leaves on liver lipid levels in Hy-Line Brown laying hens fed a high-fat diet during the late laying period;

[0026] Figure 5 To investigate the effects of Moringa leaves on the expression of genes related to liver lipid metabolism in Hy-Line Brown laying hens fed a high-fat diet during the late laying period.

[0027] in, Figure 1 Image A and B show liver tissues of Hy-Line Brown chickens fed a basal diet and a basal diet supplemented with 10% Moringa leaf, respectively; image C shows liver tissue of Hy-Line Brown chickens fed a high-fat diet; images D, E, and F show HE-stained sections of livers of Hy-Line Brown chickens fed a corresponding basal diet, a basal diet supplemented with 10% Moringa leaf, and a high-fat diet, respectively; images G, H, and I show Oil Red O-stained sections of livers of Hy-Line Brown chickens fed a corresponding basal diet, a basal diet supplemented with 10% Moringa leaf, and a high-fat diet, respectively.

[0028] Figure 2 In the middle section, A, B, and C are liver tissues of Hy-Line Brown chickens fed with high-fat diets supplemented with 2.5%, 5%, and 10% Moringa leaves, respectively; D, E, and F are HE-stained sections of livers of Hy-Line Brown chickens fed with high-fat diets supplemented with different amounts of Moringa leaves, respectively; G, H, and I are Oil Red O-stained sections of livers of Hy-Line Brown chickens fed with high-fat diets supplemented with different amounts of Moringa leaves, respectively.

[0029] Figure 3 The values ​​of ALT, AST, TC, TG, HDL, and LDL in the serum of each group are represented by AF.

[0030] Figure 4 The values ​​of AC in each group represent liver index, TG, and TC content, respectively.

[0031] Figure 5 In Figure A, the expression of liver lipid metabolism-related genes is shown in the CK, CK+10% Moringa leaf, and HFD groups. In Figure B, the expression of liver lipid metabolism-related genes is shown in the high-fat diet group and the high-fat diet group with different amounts of Moringa leaf. Detailed Implementation

[0032] To clearly illustrate the technical features of this solution, the invention will be described in detail below through specific embodiments and in conjunction with the accompanying drawings.

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of this application.

[0034] This application, through investigation of the production performance, liver tissue lesions, and serum biochemical indicators of Hy-Line Brown laying hens in the late laying period, proposes the application of Moringa leaves in the preparation of products for treating and / or alleviating fatty liver in Hy-Line Brown laying hens by improving liver lipid droplet accumulation and reducing liver damage.

[0035] The term "treatment" as used above refers to the reduction, elimination, or prevention or slowing of symptoms on a temporary or permanent basis. This term includes any treatment of a disease or condition that produces any clinically necessary or beneficial effect, including but not limited to relieving or reducing one or more symptoms, or eliminating, slowing, or stopping the progression of a disease or condition.

[0036] In the above applications, the fatty liver refers to fatty liver hemorrhage syndrome; the dosage of Moringa leaves as a feed additive is 5-10% of the feed weight; preferably 5%.

[0037] The moringa leaves mentioned above are moringa leaf powder, a commercially purchased product.

[0038] The following detailed embodiments illustrate the application of Moringa leaves in the preparation of products for treating and / or alleviating fatty liver in poultry during the late laying period.

[0039] The following embodiments are for illustrative purposes only and are not intended to limit the scope of this application. Experimental methods in the following embodiments that do not specify specific conditions should first refer to the guidelines given in this application, or may be performed according to experimental manuals or conventional conditions in the art, or by referring to experimental methods known in the art. Unless otherwise specified, all methods are conventional methods in the art.

[0040] In the following specific embodiments, unless otherwise specified, slight deviations may exist within the weighing accuracy range for the measurement parameters of the raw material components. For temperature and time parameters, acceptable deviations due to instrument testing accuracy or operational precision are permissible. The equipment and raw materials used are commercially available or commonly used in the art.

[0041] Example 1

[0042] This study provides information on the application of Moringa leaves in alleviating fatty liver in Hy-Line Brown laying hens during the late laying period. Moringa leaf powder is selected from Moringa leaves.

[0043] The fatty liver model selected in this embodiment is a high-energy, low-protein (HFD) induced fatty liver model, in which Hy-Line Grey laying hens are continuously fed a high-energy, low-protein diet for 12 weeks, during which Moringa leaf intervention is provided. Specific details are as follows:

[0044] 1.1 Experimental Animals

[0045] 450 healthy, 450-day-old Hy-Line Brown chickens with similar weight and egg production rate were raised at the Institute of Poultry Science, Shandong Academy of Agricultural Sciences.

[0046] 1.2 Main Reagents

[0047] Total cholesterol (T-CHO) assay kit (catalog number A111-1-1) and triglyceride (TG) assay kit (catalog number A110-1-1) were purchased from Nanjing Jiancheng Technology Co., Ltd.; hematoxylin-eosin staining solution (catalog number B1002), Evo M-MLV reverse transcription premixed kit (catalog number AG11728), and SYBR Green Pro Taq HS premixed qPCR kit (catalog number AG11701) were purchased from Jiangsu Aidisheng Biotechnology Co., Ltd.; 4% tissue cell fixative (catalog number P1110-500ml) and Oil Red staining solution (catalog number G1015) were purchased from Servicebio; anhydrous ethanol (catalog number 10009218) and neutral resin (catalog number 10004160) were purchased from Sinopharm Chemical Reagent Co., Ltd.; and the Simply P total RNA extraction kit (catalog number BSC52M1) for nucleic acid extraction or purification was purchased from BioFlux.

[0048] 1.3 Main Instruments

[0049] Fully automated biochemical analyzer (MINDRAY, BS-360E), dehydrator (Wuhan Junjie Electronics Co., Ltd., JJ-12J), embedding machine (Wuhan Junjie Electronics Co., Ltd., JB-P5), pathological slide machine (Shanghai Leica Instruments Co., Ltd., RM2016), tissue spreader (Zhejiang Jinhua Kedi Instrument Equipment Co., Ltd., KD-P), upright optical microscope (Nikon, Eclipse CI), high-throughput tissue homogenizer (Shanghai Jingxin Industrial Development Co., Ltd., JXFSTPRP-24), gene amplification instrument (Hangzhou Baiheng Technology Co., Ltd., GE4852T), C1000 Touch Thermal Cycler (BIO-RAD, CT014878).

[0050] 1.4 Experimental grouping and treatment

[0051] Forty-fivety healthy, 450-day-old Hy-Line Brown laying hens with similar weight and egg production rates were selected. After a 10-day acclimatization period, they were randomly divided into six treatments, with five replicates per treatment and 15 hens per replicate. The replicates were evenly distributed throughout the chicken house. The treatment groups were as follows:

[0052] The diet groups were: basal diet (CK), basal diet + 10% Moringa leaf powder (CK + MOL10), high-fat diet (HFD), high-fat diet + 2.5% Moringa leaf powder (HFD + MOL2.5), high-fat diet + 5% Moringa leaf powder (HFD + MOL5), and high-fat diet + 10% Moringa leaf powder (HFD + MOL10).

[0053] Prepare basal and high-fat diets according to the formulas in Table 1. The formal trial period was 85 days, during which the number of eggs laid and daily feed intake were recorded. The experimental hens had free access to feed and water. After the trial, 10 hens were randomly selected from each group to collect blood and separate serum. The levels of total cholesterol (TC), triglycerides (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) in the serum were measured using an automated biochemical analyzer. Liver tissue was collected after dissection of the hens. After rinsing with ice-cold physiological saline, one portion of the tissue sample was fixed in 4% formaldehyde solution, embedded in paraffin, and stained with Oil Red O for observation of tissue lesions under an optical microscope. Another portion of the tissue sample (0.2g) was weighed, rinsed in ice-cold physiological saline, dried with filter paper, placed in a beaker, and 9 times the volume of anhydrous ethanol was added. The mixture was mechanically homogenized under ice-water bath conditions, centrifuged, and the supernatant was collected. The TG and TC levels in the supernatant were measured according to the kit instructions. Total RNA was extracted from liver tissue and reverse transcribed according to the kit. The reverse transcription product was then used as a substrate for qRT-PCR assay.

[0054] Table 1. Basal and High-Fat Diet Formulations

[0055]

[0056]

[0057] Note: Each kilogram of premixed feed provides: Vitamin A 9500 IU, Vitamin D3 4000 IU, Vitamin E 25 IU, Vitamin K3 2.5 mg, Vitamin B1 2.5 mg, Vitamin B2 5.5 mg, Vitamin B6 4 mg, Vitamin B... 12 0.02mg, Biotin 0.2mg, Pantothenic Acid 8mg, Folic Acid 1mg, Choline 450mg, Iron 60mg, Copper 10mg, Manganese 110mg, Zinc 100mg, Iodine 1.0mg, Selenium 0.3mg.

[0058] 1.5 Statistical Analysis

[0059] Results were processed using SPSS 19.0. Data are expressed as mean ± standard error (x ± SEM). One-way ANOVA was used, and LSD method was used for intergroup comparisons. P < 0.05 was considered statistically significant.

[0060] 2. Application Experiment Results

[0061] 2.1 Effects of Moringa Leaves on Laying Performance of Hens Feeded on High-Fat Diets

[0062] As shown in Table 2, there were significant differences in egg production rate and average daily feed intake between the CK group and the HFD group. The egg production rate of the HFD+MOL5 and HFD+MOL10 groups was significantly higher than that of the HFD group, and the HFD+MOL5 group had the highest egg production rate, showing the best effect on improving egg production rate. The average daily feed intake of the HFD+MOL5 group was significantly higher than that of the HFD group.

[0063] Table 2. Effects of Moringa Leaf on Laying Performance of Hens Fed on High-Fat Diets in Late Laying Period

[0064]

[0065] 2.2 Effects of Moringa Leaves on Liver Tissue Lesions in Late Laying Hens Fed with High-Fat Diets

[0066] Depend on Figure 1 It can be seen that, compared with the CK group, the HFD group showed a significant increase in lipid droplets and inflammatory cells in Oil Red O stained sections (see blue and red arrows in Figures F and I, respectively), while the CK+MOL10 group showed no significant difference. Figure 2 Compared with the HFD group, the HFD groups treated with different doses of MOL significantly reduced the proportion of lipid droplets in the liver. The results suggest that HFD treatment significantly increases steatosis in the liver of laying hens, while MOL treatment significantly alleviates HFD-induced steatosis.

[0067] 2.3 Effects of Moringa Leaves on Serum Biochemical Parameters of Laying Hens Fed with High-Fat Diets

[0068] like Figure 3 As shown, compared with the CK group, the levels of AST, ALT, TC, TG, HDL, and LDL in the serum of the HFD group were significantly increased (p<0.05), while there was no significant difference in the CK+MOL10 group. Compared with the HFD group, different doses of MOL added significantly reduced the levels of AST, ALT, TC, TG, HDL, and LDL in the serum (p<0.05), with the TG level significantly reduced in the HFD+MOL5 and HFD+MOL10 groups, and the reduction was most significant in the HFD+MOL5 group.

[0069] 2.4 Effects of Moringa Leaves on Liver Lipid Indicators in Laying Hens Fed a High-Fat Diet

[0070] like Figure 4 As shown, compared with the CK group, the HFD group had significantly higher liver index and liver TG content, while there was no significant difference in CK+MOL10. Compared with the HFD group, the HFD+MOL5 and HFD+MOL10 groups had significantly lower liver index, and the HFD+MOL2.5, HFD+MOL5, and HFD+MOL10 groups had significantly lower TG content. There was no significant difference in liver TC content among the groups.

[0071] 2.5 Effects of Moringa Leaf on the Expression of Lipid Metabolism-Related Genes in the Liver of Laying Hens Fed a High-Fat Diet

[0072] like Figure 5 As shown, compared with the CK group, the HFD group showed significantly decreased PPARα mRNA expression and significantly increased PPARγ, FABP1, and ACC expression, while the CK+MOL10 group showed significantly decreased FAS expression; there was no significant change in SREBP1-c among the groups. Compared with the HFD group, the HFD+MOL5 group showed significantly increased PPARα mRNA expression, the HFD+MOL10 group showed significantly decreased PPARγ expression, and the MOL groups at each dose showed significantly decreased SREBP1-c, FABP1, ACC, and FAS expression.

[0073] The above results indicate that adding the optimized dosage of Moringa leaves as described in this invention has a significant alleviating and improving effect on fatty liver in Hy-Line Brown laying hens during the later stages of egg production. It can reduce fatty liver and blood lipid levels in Hy-Line Brown laying hens during the later stages of egg production, increase antioxidant activity, significantly affect the production performance of laying hens during the later stages of egg production, and slow down the decline in egg production rate.

[0074] The above provides a detailed description of the application of Moringa leaves provided by this invention in the preparation of products for treating and / or alleviating fatty liver in poultry during the late laying period. The specific embodiments described above should not be construed as limiting the scope of protection of this invention. Any alternative modifications or variations made to the embodiments of this invention by those skilled in the art will fall within the scope of protection of this invention.

[0075] Any aspects of this invention not described in detail are well-known to those skilled in the art.

Claims

1. The application of Moringa leaves in the preparation of products for treating and / or alleviating fatty liver in late-laying poultry, characterized in that, The poultry in question are Hy-Line Brown laying hens in their late egg-laying stage.

2. The application according to claim 1, characterized in that, The fatty liver mentioned is fatty liver hemorrhage syndrome; the dosage of Moringa leaves as an additive is 5-10% of the feed weight.

3. The application according to claim 2, characterized in that, Moringa leaves were added at a rate of 5%.

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