Feed additive for improving fallopian tube inflammation and eggshell quality of laying hens in later laying period and application of feed additive

Abstract

The invention relates to the technical field of laying hen breeding feed, particularly discloses a feed additive for improving salpingitis and eggshell quality of laying hens in the later egg laying period and application of the feed additive, and aims to relieve the salpingitis of the laying hens in the later egg laying period and improve production performance and egg quality. The technical key points are as follows: when 200 mg / kg of ME or an equal amount of Rottlerin is added into the feed, mucosal lesion and inflammatory cell infiltration caused by salpingitis can be effectively relieved; specifically, swelling and bleeding of the fallopian tube are relieved, epithelial resistance of mucous membranes is improved, expression of tight junction protein is increased, and the level of inflammatory factors is reduced, so that the pathological state of the fallopian tube is remarkably improved; meanwhile, the additive significantly improves the laying rate and egg quality of the laying hens, especially the ultrastructure and strength of eggshells, reduces the feed-egg ratio, and optimizes the production performance of the laying hens. The additive disclosed by the invention can relieve inflammatory response, improve the health level of laying hens and reduce the use of medicines to a certain extent, so that the safe production of the laying hens is guaranteed, and the additive has important application value.

Description

Technical Field

[0001] This invention relates to the field of egg-laying hen feed technology, and more specifically, to a feed additive that improves oviduct inflammation and eggshell quality in late-laying hens and its application. Background Technology

[0002] Oviductitis is a common and serious disease in intensive egg-laying hen farming, mainly manifested as oviduct infection and inflammation, often caused by bacterial, viral, or fungal infections. Affected hens frequently experience decreased egg production, increased defective eggs, or even cessation of egg production, severely impacting production performance and economic benefits. Currently, adding antibiotics to feed is the primary means of preventing and treating oviductitis; however, with the complete ban on antibiotics in feed, traditional prevention and treatment methods face severe challenges, making the search for safe and effective alternatives an urgent priority.

[0003] *Mallotus philippensis*, a perennial shrub or small tree belonging to the Euphorbiaceae family, is widely distributed in Sichuan, Yunnan, Guizhou, Hubei, Jiangxi, and other regions of my country and is an important medicinal plant. Rottlerin, extracted from its fruit, is a natural plant polyphenol compound with significant anti-infective, anti-inflammatory, and antioxidant biological activities, showing good potential in regulating the body's immune function and alleviating inflammatory responses.

[0004] Therefore, this application has developed a novel feed additive based on the extract of *Rhizoctonia solani* and its active ingredient Rottlerin, providing a feed additive and its application to improve oviduct inflammation and eggshell quality in laying hens during the late laying period, in order to solve the above problems. This is of great significance for replacing antibiotics and effectively preventing and treating oviduct inflammation in laying hens. Summary of the Invention

[0005] The purpose of this invention is to solve the technical problems mentioned in the background section and to provide a feed additive that improves oviduct inflammation and eggshell quality in laying hens during the late laying period, and its application. The feed additive of this invention can alleviate oviduct inflammation in laying hens, improve their immunity and resistance, improve production performance, reduce drug use and residues, and ensure safe production of laying hens.

[0006] The above-mentioned objective of the present invention is achieved as follows: One aspect of the present invention is to provide a feed additive that improves oviduct inflammation and eggshell quality in laying hens during the late laying period, wherein the feed additive is a extract of *Rhizoctonia solani* or the active ingredient of *Rhizoctonia solani*, Rottlerin.

[0007] Furthermore, the dosage of the *Rhizoctonia solani* extract is 200 mg / kg; the dosage of the active ingredient Rottlerin from *Rhizoctonia solani* is 5.26 mg / kg.

[0008] In this program, the additive is added to the feed and fed to laying hens infected with oviductitis. This can effectively alleviate the decline in production performance and egg quality caused by inflammation, reduce oviduct damage and inflammatory response, and improve the immunity and resistance of laying hens to oviductitis.

[0009] Another aspect of the present invention provides a layer hen feed based on the aforementioned additives, the feed comprising a basal feed and the aforementioned additives. Feeding this feed to layer hens infected with oviduct inflammation can effectively alleviate the decline in production performance and egg quality caused by inflammation, reduce oviduct damage and inflammatory response, and improve the immunity and resistance of layer hens to oviduct inflammation.

[0010] Furthermore, the basic feed consists of the following components, expressed as a percentage of dry matter content: Corn 64%, soybean meal 25%, limestone powder 4.2%, corn gluten meal 5%, premix 1.8%; The premixed feed, calculated per kilogram of complete feed, consists of the following components: VA 12500 IU, VD3 4125 IU, VE 15 IU, VK3 2 mg, VB1 25 mg, VB2 8.5 mg, biotin 2 mg, choline 500 mg, niacin 32.5 mg, manganese 65 mg, zinc 65 mg, iron 60 mg, copper 8 mg, iodine 1 mg, and selenium 0.30 mg.

[0011] The present invention also provides the application of the above-mentioned additive or the above-mentioned layer hen feed in late-laying layer hen farming. In this invention, by adding the additive to the layer hen feed, the immunity and disease resistance of the layer hen can be improved, especially its resistance to oviductitis. Furthermore, feeding the layer hen with this feed can improve its immunity and disease resistance, especially its resistance to oviductitis.

[0012] The present invention also provides the application of the above-mentioned additives in the preparation of products that enhance the resistance of laying hens to oviduct inflammation, or in the preparation of products that enhance the immunity of laying hens, or in the preparation of products that alleviate oviduct inflammation in laying hens.

[0013] Among them, the feed additive can be prepared into corresponding products, such as feed, and applied to the breeding of laying hens, which can improve the resistance of laying hens to oviduct inflammation.

[0014] In the application of feed additives in the preparation of products that enhance the immunity of laying hens, the feed additive is either 200 mg / kg of coarse bran extract or 5.26 mg / kg of Rottlerin. Therefore, by preparing corresponding products, such as feed, using this feed additive and applying it to the raising of laying hens, the immunity and resistance of the hens can be improved.

[0015] In the application of feed additives in the preparation of products that alleviate oviduct inflammation in laying hens, the feed additive is either 200 mg / kg of coarse bran extract or 5.26 mg / kg of Rottlerin. Thus, corresponding products, such as feed, are prepared using this feed additive and applied to the raising of laying hens to alleviate and reduce the impact of inflammation on them.

[0016] The present invention also provides the application of the above-mentioned additive in the preparation of products that improve the abnormal production performance of laying hens caused by oviductitis, wherein the production performance includes average daily weight gain, laying rate, egg weight, and feed conversion ratio. By preparing corresponding products, such as feed, using this feed additive and applying it to the raising of laying hens, the loss of production performance due to inflammation can be alleviated, specifically manifested in improved production performance, thereby mitigating and reducing the impact of inflammation on laying hens.

[0017] The present invention also provides the application of the above-mentioned additive in the preparation of products that improve egg quality abnormalities caused by oviductitis in laying hens, wherein the egg quality includes Haugh unit, eggshell thickness, and eggshell strength. By preparing corresponding products, such as feed, using this feed additive and applying it to the raising of laying hens, the loss of egg quality due to inflammation can be alleviated, specifically manifested in the improvement of egg quality abnormalities, thereby mitigating and reducing the impact of inflammation on laying hens.

[0018] The present invention also provides the application of the above-mentioned additive in the preparation of products that alleviate oviduct damage caused by oviductitis in laying hens, wherein the oviduct damage includes abnormalities in one or more of the following indicators: intermucosal vacuoles and inflammatory cell infiltration. By preparing corresponding products, such as feed, using this feed additive and applying it to the raising of laying hens, the damage to the oviduct caused by inflammation can be alleviated, thereby mitigating and reducing the impact of inflammation on laying hens.

[0019] The present invention also provides the application of the above-mentioned additive in the preparation of products that inhibit inflammatory factors caused by oviductitis in laying hens, wherein the inflammatory factors include any one or more of IL-1β, IL-6, and TNF-α. By preparing corresponding products, such as feed, from this feed additive and applying them to the raising of laying hens, the effects of inflammation on laying hens can be alleviated and reduced by inhibiting inflammatory factors.

[0020] Compared with the prior art, the present invention has the following beneficial effects: 1. The additive provided by the present invention is a rough straw extract or its active ingredient Rottlerin, which is added to the feed of laying hens infected with oviductitis (treated with lipopolysaccharide modeling); studies have shown that the additive can effectively alleviate the decline in production performance, reduced egg quality, oviduct damage and inflammatory response caused by oviductitis, and significantly improve the resistance of laying hens to oviductitis. 2. The additives provided by this invention, namely, the extract of *Rhizoctonia solani* and Rottlerin, can significantly inhibit the expression of inflammatory factors, improve the oviduct mucosal structure, and repair damaged tissues. Therefore, by inhibiting the inflammatory response, this additive can effectively alleviate the symptoms of oviductitis in laying hens, improve the overall health of laying hens, and to a certain extent reduce drug use, reduce antibiotic dependence, and ensure the safety of laying hen production, thus having important application value. Attached Figure Description

[0021] Figure 1 This is a statistical result chart of production performance in the embodiments of the present invention (CON represents the control group, LPS represents the inflammation blank group, LPS+ME represents the inflammation Caulis Branbarb Extract group, LPS+Rottlerin represents the inflammation Rottlerin group, the same letter indicates no significant difference (p>0.05), different letters indicate significant difference (). P <0.05); Figure 2 This is a statistical result chart of egg quality in an embodiment of the present invention (CON represents the control group, LPS represents the inflammation blank group, LPS+ME represents the inflammation coarse bran extract group, LPS+Rottlerin represents the inflammation Rollerin group, and the same letter indicates no significant difference). P >0.05), different letters indicate significant differences ( P <0.05); Figure 3 These are pathological images of fallopian tube injury in embodiments of the present invention (CON represents the control group, LPS represents the inflammation blank group, LPS+ME represents the inflammation *Cinnamomum camphora* extract group, LPS+Rottlerin represents the inflammation Rollerin group, and the same letter indicates no significant difference). P >0.05), different letters indicate significant differences ( P <0.05); Figure 4 This is a statistical chart of inflammatory factor content in the embodiments of the present invention (CON represents the control group, LPS represents the inflammatory blank group, LPS+ME represents the inflammatory *Cinnamomum camphora* extract group, LPS+Rottlerin represents the inflammatory Rollerin group, and the same letter indicates no significant difference). P >0.05), different letters indicate significant differences ( P <0.05); Figure 5This invention relates to the effects of dietary supplementation with *Branchella esculenta* extract on oviduct inflammatory response in laying hens (A: oviduct inflammatory cytokine levels; B: serum inflammatory cytokine levels; C: oviduct inflammatory cytokine transcription levels; D: CD4+ and CD8+ gene transcription levels, n=3; E: serum and oviduct LPS levels, n=8; F: oviduct CD4+ fluorescence signal; G: average CD4+ fluorescence intensity, n=3). Figure 6 This invention relates to the effects of dietary supplementation with *Rhizoctonia solani* extract on the expression of key proteins in the oviduct PKC family and NF-κB signaling pathway (A: oviduct PKCδ fluorescence signal; B: average PKCδ fluorescence intensity; C: PKC protein transcription level in the oviduct, n=6; D: expression of key proteins in the PKCδ / NF-κB signaling pathway; E: protein grayscale analysis, n=3). Figure 7 In this embodiment of the invention, *Branchella esculenta* extract and Rotterin alleviate the reduction in feed intake, egg production rate, and egg quality in laying hens caused by LPS stimulation. In the figure: LPS / PBS represents oviduct perfusion with LPS or PBS on day 14, where CON = control group; LPS = LPS oviduct perfusion group; ME+LPS = *Branchella esculenta* extract + LPS oviduct perfusion group; Rotterin+LPS = Rotterin + LPS oviduct perfusion group (A: Average daily feed intake (ADFI); B: Egg production rate (LR); C: Average egg weight (AEW); D: Haugh unit; E: Eggshell strength; F: Eggshell thickness, n=6). Figure 8 This invention relates to the effects of dietary supplementation with *Rhizoctonia solani* extract and Rotterin on the oviduct health of laying hens under LPS stimulation (A: changes in the outer and inner surfaces of the oviduct; B: oviduct lesion score; C: weight per unit length of oviduct; D: oviduct mucosal resistance). Figure 9 This invention relates to the effects of dietary supplementation with *Rhizoctonia solani* extract and Rottlerin on oviduct function in laying hens stimulated by LPS (A: oviduct CD4+, PCNA, and Claudin-1 fluorescence signals; B: average CD4+ fluorescence intensity; C: average CAN fluorescence intensity; D: average Claudin-1 fluorescence intensity, n=3). Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0023] The implementation of the present invention will be described in detail below with reference to specific embodiments.

[0024] Reference Figures 1-9 The image shown is a preferred embodiment of the present invention.

[0025] Example: The present invention provides a feed additive for improving oviduct inflammation and eggshell quality in laying hens during the late laying period, and its application. The aim is to develop a novel feed additive based on *Rhizoctonia solani* extract and its active ingredient Rottlerin to replace antibiotics and effectively prevent oviduct inflammation in laying hens.

[0026] The specific implementation experiments of this invention are as follows: 1. Test subjects The experimental animals were 350-day-old Hy-Line Brown chickens, purchased from Suicheng Qinwang Poultry Farm in Suixi County, Guangdong Province.

[0027] 2. Modeling of oviductitis in laying hens A lipopolysaccharide (LPS) model of oviductitis in laying hens was established using the following method: One person held the hen upright with its head down and cloaca up, gently pressing down on both sides of the cloaca while the other hand supported the hen's chest to allow the oviduct opening to protrude; another person injected the prepared LPS into the uterine portion of the oviduct. After injection, the hen was placed in a cage and observed for 24 hours. Successful model establishment was indicated by swollen anus, discharge of yellowish-white feces staining the perianal feathers, and the production of hemorrhagic eggs.

[0028] 3. Experimental Design and Grouping Seventy-two healthy, late-laying hens of similar size were randomly selected and divided into a blank control group (n=18) and a model group. The 54 hens in the model group underwent lipopolysaccharide (LPS) treatment to induce an inflammation model. These 54 hens were then randomly divided into three groups: a LPS control group (inflammation blank group), a 200 mg / kg *Rhizoctonia solani* extract group (inflammation *Rhizoctonia solani* extract group), and a 5.26 mg / kg Rottlerin group (inflammation Rottlerin group). The blank control group and the LPS control group (inflammation blank group) were fed a basal diet. The 200 mg / kg *Rhizoctonia solani* extract group (inflammation *Rhizoctonia solani* extract group) was fed a basal diet supplemented with 200 mg / kg *Rhizoctonia solani* extract. The 5.26 mg / kg Rottlerin group (inflammation Rottlerin group) was fed a basal diet supplemented with 5.26 mg / kg Rottlerin. The hens were cage-housed with free access to feed and water for 16 days. The composition and nutrient levels of the basal diet are shown in Tables 1 and 2. On day 17, six animals from each group were selected for slaughter and sampling.

[0029] Table 1. Composition of the basic diet project content(%) raw material corn 64.00 soybean meal 25.00 stone powder 4.20 Corn protein powder 5.00 <![CDATA[Premix 1 > 1.80 The premixed feed consists of (calculated per kilogram of complete feed): VA 12500 IU, VD3 4125 IU, VE 15 IU, VK3 2 mg, VB1 25 mg, VB2 8.5 mg, biotin 2 mg, choline 500 mg, niacin 32.5 mg, manganese 65 mg, zinc 65 mg, iron 60 mg, copper 8 mg, iodine 1 mg, and selenium 0.30 mg.

[0030] Table 2 Nutritional Levels of Basal Diets project content(%) Nutritional level Metabolizable energy (MJ / kg) 11.29 crude protein 17.47 Coarse ash 11.25 Crude fat 2.20 calcium 3.60 Total phosphorus 0.54 Non-phytic acid phosphorus 0.35 Lysine 0.75 Methionine + Cysteine 0.74 4. Feeding and Management During the experiment, chickens were cage-raised with 16 hours of light per day. The temperature inside the chicken house was controlled at around 25℃, and the relative humidity was around 77%. They were fed artificially and had free access to water. The experiment lasted for 19 days, including a 3-day pre-feeding period, a 2-day modeling period, and a 14-day formal experiment. The condition of the flock was observed daily.

[0031] 5. Measurement Indicators and Methods 5.1 Production Performance Measurement Record the daily egg production count and total daily egg weight in repeated batches. Calculate the feed consumption of the chickens weekly. Weigh the laying hens at the beginning and end of the experiment. The formulas for calculating the various production performance indicators are as follows: (1) Egg production rate = number of eggs produced per day / number of animals in stock; (2) Average daily feed intake = total feed consumption / number of days in the trial / number of animals in stock; (3) Average egg weight = total egg weight / number of eggs laid; (4) Feed-to-egg ratio = total feed consumption / total egg weight.

[0032] 5.2 Egg Quality Determination Four eggs are randomly selected from each replicate, labeled with the group number and laying date, for determining egg quality. The calculation formula is as follows: (1) Haugh unit = 100lg(H-1.7W0.37+7.57), where: H represents the height of concentrated albumen (mm); W represents the weight of the albumen (g); (2) Eggshell thickness = (thickness at blunt end + thickness at middle end + thickness at sharp end) / 3; (3) Eggshell strength = pressure on the eggshell when it breaks / area of ​​the eggshell under pressure.

[0033] 5.3 Histopathological observation The oviduct and uterus of laying hens were taken, fixed in 4% paraformaldehyde for one week, embedded in paraffin, sectioned (4 μm), stained with hematoxylin and eosin (HE), and examined under a microscope to observe pathological changes. Another tissue was taken, fixed in glutaraldehyde, embedded in epoxy resin, ultrathin sectioned (70 nm), and double-stained. The epithelial ultrastructure was observed by transmission electron microscopy to assess cell integrity and inflammatory infiltration (SC: secretory epithelial cells; L: lymphocytes).

[0034] 5.4 Measurement of serum inflammatory markers Tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were all obtained from Nanjing Jiancheng Biotechnology Institute.

[0035] 6. Statistical Analysis Experimental data were organized using Excel and analyzed using SPSS 26.0 (IBM, USA). One-way ANOVA and Kruskal-Wallis tests were used to analyze normally and non-normally distributed data, respectively. Multiple comparisons were performed using either the parametric Tukey test or the non-parametric Tamhane test.

[0036] 7. Results Analysis 7.1 Effects of Coarse Rice Bran Extract or Rotterin on Laying Hen Production Performance Oviduct perfusion with lipopolysaccharide successfully induced oviductitis, significantly reducing the average daily feed intake, laying rate, and average egg weight in laying hens, and increasing the feed conversion ratio (FCR). Figure 1 AD in P <0.05); the addition of coarse bran extract or Rollerin to the diet can significantly reverse this decline in production performance.

[0037] 7.2 Effects of Coarse Rice Bran Extract or Rotterin on Egg Quality Oviduct perfusion with lipopolysaccharide successfully induced oviductitis and significantly reduced Haugh units in eggs produced by laying hens in the late laying period. Figure 2 A in P <0.05), eggshell thickness ( Figure 2 B in P <0.01) and eggshell strength ( Figure 2 C in P Egg quality can be significantly improved by adding coarse bran extract or Rollerin to the diet (<0.01).

[0038] 7.3 Effects of *Branch husk extract* or Rottlerin on the morphology of oviduct tissue in laying hens Depend on Figure 3 As indicated by A in the text, after HE staining of sections of the oviduct and uterus of laying hens, microscopic observation revealed varying degrees of oviduct mucosal damage and increased mucosal spaces in the inflammatory blank group, the inflammatory *Branchella esculenta* extract group, and the inflammatory Rottlerin group. Transmission electron microscopy showed the following results: Figure 3 In B, lipopolysaccharide perfusion induces oviductitis, causing swelling of secretory epithelial cells in chicken oviducts, their shape to evolve into irregular polygons, chromatin aggregation and marginalization of cell nuclei, and a large number of dark lymphocytes infiltrating the submucosa; the addition of coarse bran extract and Rotterin can restore the normal microscopic morphology and structure of oviduct epithelial cells (SC: secretory epithelial cells; L: lymphocytes).

[0039] 7.4 Effects of *Rhizoctonia solani* extract or Rotterin on inflammatory factors in laying hens Interleukin-1β (IL-1β): Compared with the control group, the concentration of the inflammatory factor IL-1β was significantly increased in the blank inflammation group. Figure 4 A in P <0.01), indicating that the inflammatory response increases the concentration of IL-1β in laying hens, and inflammation has a significant impact on the immune marker IL-1β in laying hens; while the concentration of IL-1β in the inflammatory *Brassica napus* extract group or the inflammatory Rottlerin group was significantly lower than that in the inflammatory blank group. P <0.01), its concentration was slightly higher than that of the control group ( P >0.05), indicating that the extract of *Rhizoctonia solani* or Rotterin can significantly alleviate the effects of inflammation on laying hens and improve their immunity and resistance to inflammation.

[0040] Interleukin-6 (IL-6): Compared with the control group, the concentration of the inflammatory factor IL-6 was significantly increased in the blank inflammation group. Figure 4 B in P<0.01), indicating that the inflammatory response increases the concentration of IL-6 in laying hens, and inflammation has a significant impact on the IL-6 immune marker in laying hens; while the IL-6 concentration in the inflammatory *Brassica napus* extract group or the inflammatory *Rottlerin* group was significantly lower than that in the inflammatory control group. P <0.01), its concentration was slightly higher than that of the control group ( P >0.05), indicating that the extract of *Rhizoctonia solani* or Rotterin can significantly alleviate the effects of inflammation on laying hens and improve their immunity and resistance to inflammation.

[0041] Tumor necrosis factor α (TNF-α): Compared with the control group, the concentration of the inflammatory factor TNF-α was significantly increased in the blank inflammation group. Figure 4 C, P <0.01 indicates that the inflammatory response increases the concentration of TNF-α in laying hens, and inflammation has a significant impact on the immune indicator TNF-α in laying hens; while the concentration of TNF-α in the inflammatory *Cinnamomum camphora* extract group or the inflammatory *Rottlerin* group was significantly lower than that in the inflammatory blank group. P <0.01), its concentration was slightly higher than that of the control group ( P >0.05), indicating that the extract of *Rhizoctonia solani* or Rotterin can significantly alleviate the effects of inflammation on laying hens and improve their immunity and resistance to inflammation.

[0042] The above results indicate that *Branchella esculenta* extract or Rotttlerin exhibits significant effects in reducing the expression of inflammatory factors (such as TNF-α, IL-1β, and IL-6), particularly by inhibiting the inflammatory response through regulation of the NF-κB signaling pathway. The anti-inflammatory effect of *Branchella esculenta* extract is closely related to its active ingredient, Rotttlerin, which can effectively alleviate tissue damage caused by inflammation, thus mitigating the impact of inflammation on laying hens and enhancing their immunity and resistance to inflammation.

[0043] 7.5. Extracts from *Branch husk* inhibit inflammatory cell infiltration and significantly reduce oviduct inflammation in laying hens. Compared with the control group, dietary supplementation with different concentrations of *Rhizoma Citri Reticulatae* extract significantly reduced the levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) in the fallopian tube tissue. Figure 5 A in P <0.05%, the addition of 400 mg / kg of *Branchella esculenta* extract significantly increased the levels of the anti-inflammatory cytokine IL-4 in the fallopian tubes and serum. Figure 5 AB in P <0.05; In addition, the transcriptional levels of inflammatory cytokines (IL-1β, IL-6) and T lymphocyte marker proteins (CD4+, CD8+) in the fallopian tubes of the *Corydalis yanhusuo* group were significantly downregulated ( Figure 5CD in P <0.05); Adding different concentrations of *Branchella esculenta* extract to the diet significantly reduced LPS levels in the fallopian tubes and serum ( Figure 5 E in P <0.05); Immunofluorescence staining results of CD4+ T lymphocyte expression in the fallopian tube were consistent with qPCR results. Figure 5 FG in P <0.05).

[0044] 7.6. *Branch bran* extract participates in the regulation of the NF-κB signaling pathway via PKCδ. Depend on Figure 6 According to AC, compared with the control group, dietary supplementation with *Branchella esculenta* extract inhibited the transcription level of PKCδ in a concentration-dependent manner. P <0.05; Adding different concentrations of *Branchella esculenta* extract to the diet could downregulate the expression levels of key proteins in the NF-κB signaling pathway, p-IKB, IKB, p-p65, and p65, as well as their downstream proteins IL-1β and IL-6. Figure 6 DE in P <0.05).

[0045] 7.7 Effects of *Branchella esculenta* extract and its active ingredient, Rottlerin, on the production performance and egg quality of laying hens with LPS-induced oviductitis. like Figure 7 As shown, compared with the CON group, oviduct perfusion with LPS significantly reduced the average daily feed intake and egg production rate of laying hens. Figure 7 AB in P <0.05); the addition of coarse bran extract and Rottlerin to the diet can significantly reverse this decline in production performance ( Figure 7 AB in P <0.05); In addition, LPS-induced oviduct inflammation significantly reduced the average egg weight of eggs laid by the experimental chickens ( P <0.05), Hardy units ( P <0.05), eggshell strength ( P <0.01) and eggshell thickness ( P <0.01%, dietary supplementation with coarse bran extract and Rottlerin significantly reversed the decrease in egg quality induced by LPS perfusion in the oviduct ( Figure 7 CF in P <0.05).

[0046] 7.8. The alleviating effect of *Branchella esculenta* extract and its active ingredient, Rottlerin, on oviduct inflammation in laying hens stimulated by LPS. 7.8.1. Coarse bran extract and Rotterin alleviate LPS-induced oviduct lesions in chickens. like Figure 8 As shown, compared with the control group, LPS perfusion caused oviduct edema and hemorrhage in chickens, leading to oviduct mucosal lesions. Figure 8 (A in the text); while feeding with coarse bran extract and Rotterin can significantly reduce LPS-induced fallopian tube lesions ( Figure 8 B in P <0.01); meanwhile, in the LPS group, fallopian tube edema led to a significant decrease in the weight per unit length of fallopian tube and the mucosal resistance of the fallopian tube, while extracts of *Branchella esculenta* and Rottlerin significantly increased the weight per unit length of fallopian tube and the mucosal resistance of the fallopian tube. Figure 8 CD in P <0.01).

[0047] 7.8.2. Coarse rice bran extract and Rotterin alleviate LPS-induced inflammatory cell infiltration and functional impairment of chicken oviducts. like Figure 9 As shown, immunofluorescence was used to detect the expression of CD4+ T lymphocyte marker protein CD4+, cell proliferation marker protein PCNA, and barrier marker protein Claudin-1 in the fallopian tube. Compared with the control group, LPS perfusion significantly increased T lymphocyte infiltration in the fallopian tube tissue and significantly inhibited fallopian tube epithelial cell proliferation and mucosal barrier function. Figure 9 AD in P <0.01); Coarse rice bran extract and Rotterin can reduce lymphocyte infiltration in chicken oviduct tissue after LPS perfusion, promote oviduct epithelial cell proliferation and repair its barrier function.

[0048] In summary, the above-described experimental results demonstrate the potential of *Rhizoctonia solani* extract and its active ingredient, Rotterin, as a feed additive in alleviating oviductitis in laying hens. Its anti-inflammatory and immunomodulatory effects significantly reduce oviduct inflammation, improve the health of laying hens, and enhance production performance and egg quality. *Rhizoctonia solani* extract and Rotterin, as natural, safe, and effective alternatives, hold promise for providing new solutions to reduce antibiotic use and ensure food safety.

[0049] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A feed additive for improving oviduct inflammation and eggshell quality in laying hens during the late laying period, characterized in that, The feed additive is a extract of *Rhizoctonia solani* or the active ingredient of *Rhizoctonia solani*, Rotttlerin.

2. The feed additive according to claim 1, characterized in that, The dosage of the extract of *Rhizophora stylosa* is 200 mg / kg; the dosage of the active ingredient Rottlerin from *Rhizophora stylosa* is 5.26 mg / kg.

3. A layer hen feed based on the additive of claim 1 or claim 2, characterized in that, The feed consists of a base feed and the aforementioned additives.

4. The laying hen feed according to claim 3, characterized in that, The basic feed consists of the following components, expressed as a percentage of dry matter content: Corn 64%, soybean meal 25%, limestone powder 4.2%, corn gluten meal 5%, premix 1.8%; The premixed feed, calculated per kilogram of complete feed, consists of the following components: Composition: Vitamin A 12500 IU, Vitamin D3 4125 IU, Vitamin E 15 IU, Vitamin K3 2 mg, Vitamin B1 25 mg, Vitamin B2 8.5 mg, Biotin 2 mg, Choline 500 mg, Niacin 32.5 mg, Manganese 65 mg, Zinc 65 mg, Iron 60 mg, Copper 8 mg, Iodine 1 mg, Selenium 0.30 mg.

5. The application of the additive according to any one of claims 1-2 or the layer hen feed according to claim 3 in the late-laying stage of layer hen farming.

6. The use of the additive according to any one of claims 1-2 in the preparation of products that enhance the resistance of laying hens to oviduct inflammation, or in the preparation of products that enhance the immunity of laying hens, or in the preparation of products that alleviate oviduct inflammation in laying hens.

7. The use of the additive according to any one of claims 1-2 in the preparation of a product for improving the abnormal production performance of laying hens caused by oviductitis, characterized in that, The production performance includes average daily weight gain, egg production rate, egg weight, and feed conversion ratio.

8. The use of the additive according to any one of claims 1-2 in the preparation of a product for improving egg quality abnormalities caused by oviductitis in laying hens, characterized in that, The egg quality includes the egg's ultrastructure, Hardy units, shell thickness, and shell strength.

9. The use of the additive according to any one of claims 1-2 in the preparation of a product for alleviating oviduct damage caused by oviductitis in laying hens, characterized in that, The fallopian tube damage mentioned includes abnormalities in one or more of the following indicators: intermucosal vacuoles, inflammatory cell infiltration, etc.

10. The use of the additive according to any one of claims 1-2 in the preparation of a product that inhibits inflammatory factors caused by oviductitis in laying hens, characterized in that, The inflammatory factors include any one or more of IL-1β, IL-6, and TNF-α.

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