A composition, microemulsion for regulating the intestinal health of livestock and poultry, and a preparation method and use thereof

Abstract

The present application belongs to the field of biotechnology, and discloses a composition for regulating the intestinal health of livestock and poultry, which comprises black soldier fly larva intestinal extract, and further comprises poplar flower extract, dihydromyricetin and pine needle powder extract; the weight ratio of the black soldier fly larva intestinal extract, the poplar flower extract, the dihydromyricetin and the pine needle powder extract is 1:0.1-10:0.1-0.5:0.1-10. The composition can improve the intestinal health and reduce the diarrhea of weaned piglets; the composition of the present application is not only beneficial to the improvement of the intestinal health of piglets, but also has a good intestinal health improvement effect on pigs of other ages; in addition, the present application further provides a microemulsion containing the composition, a preparation method and use thereof.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a composition, microemulsion, preparation method, and uses for regulating the intestinal health of livestock and poultry. Background Technology

[0002] Chinese invention patents ZL201910055164.7, 202110218005.1, and 201811428762.6, proposed by Rong Ting et al., started their research with the intestinal extract of black soldier fly larvae. They extensively explained the antibacterial properties of the black soldier fly larvae intestinal extract itself, its use as a topical spray, and its use as an oral additive. Through a large number of experiments, they proved that the black soldier fly larvae intestinal extract can be effectively compounded with various ingredients to improve intestinal health, inhibit harmful bacteria, and promote the growth of beneficial bacteria.

[0003] The problem this case aims to solve is: how to improve the gut health of pigs and reduce the incidence of intestinal diseases by optimizing product formulation. Summary of the Invention

[0004] The purpose of this invention is to provide a composition for regulating the intestinal health of livestock and poultry, which can improve intestinal health and reduce diarrhea in weaned piglets and chickens; the composition of this invention is not only beneficial to the intestinal health of piglets and chickens, but also has a good effect on improving the intestinal health of pigs of other ages.

[0005] In addition, the present invention also provides microemulsions, methods for their preparation, and uses.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a composition for regulating the intestinal health of livestock and poultry, comprising black soldier fly larvae intestinal extract, and further comprising poplar flower extract, dihydromyricetin, and pine needle powder extract; wherein the weight ratio of the black soldier fly larvae intestinal extract, poplar flower extract, dihydromyricetin, and pine needle powder extract is 1:0.1-10:0.1-0.5:0.1-10.

[0007] In the above-mentioned composition for regulating the intestinal health of livestock and poultry, the weight ratio of the black soldier fly larvae intestinal extract, poplar flower extract, dihydromyricetin, and pine needle powder extract is 1:0.5-3:0.1-0.3:0.5-3.

[0008] In the above-mentioned composition for regulating the intestinal health of livestock and poultry, the weight ratio of the black soldier fly larvae intestinal extract, poplar flower extract, dihydromyricetin, dihydromyricetin, and pine needle powder extract is 1:1-2:0.2-0.3:1-2.

[0009] In the above-mentioned composition for regulating the intestinal health of livestock and poultry, the black soldier fly larvae intestinal extract contains 18.0 wt% to 22.0 wt% oleic acid glycerides, 7.7 wt% to 10.0 wt% lauric acid, and 15.5 wt% to 18.0 wt% myristic acid.

[0010] Meanwhile, the present invention also provides a microemulsion that is beneficial to the intestinal health of livestock and poultry, comprising an oil phase, an aqueous phase, a composition as described above, and an emulsifier;

[0011] The emulsifier comprises 5-8% of the total weight of the microemulsion;

[0012] The composition comprises 1-5% of the total weight of the microemulsion;

[0013] The oil phase comprises 10-15% of the total weight of the microemulsion.

[0014] In the aforementioned microemulsion beneficial to the intestinal health of livestock and poultry, the emulsifier is composed of soybean lecithin, glyceryl monolaurate, and Tween 80 in a weight ratio of 1:2:7.

[0015] The aforementioned microemulsions that are beneficial to the intestinal health of livestock and poultry also include oil-soluble vitamins, water-soluble vitamins, and amino acid compositions.

[0016] The amount of each of the oil-soluble vitamins, water-soluble vitamins, and amino acid composition shall not exceed 1% of the total weight of the microemulsion.

[0017] In the aforementioned microemulsions beneficial to the intestinal health of livestock and poultry, the oil-soluble vitamins include vitamin A, vitamin D, vitamin E, and vitamin K; the weight ratio of vitamin A, vitamin D, vitamin E, and vitamin K is 1:0.1-10:0.1-10:0-0.1.

[0018] The water-soluble vitamins include B vitamins and vitamin C; the ratio of B vitamins to vitamin C is 1:2-5.

[0019] The amino acid composition contains 1-5% threonine, 1-5% arginine, and 1-5% glutamic acid.

[0020] Finally, the present invention also provides a method for preparing the microemulsion and its uses. Specifically, the method for preparing the microemulsion is as follows: an oil-soluble component is added to an oil phase, a water-soluble component is added to an aqueous phase, the oil phase and the emulsifier are mixed uniformly, and finally the aqueous phase is added and sheared and dispersed to obtain an oil-in-water microemulsion.

[0021] The microemulsion is used to add 1-10 kg of the microemulsion per ton of drinking water to livestock and poultry drinking water.

[0022] Compared with the prior art, the beneficial effects of the present invention are:

[0023] 1. Black soldier fly larvae intestinal extract has good antibacterial effects and is effective in preventing and treating diarrhea. Poplar flower extract mainly contains flavonoids, organic acids, amino acids, sugars, and alkaloids, which can directly inhibit pathogens, improve the body's resistance, increase the tension of gastrointestinal muscles and sphincter muscles in animals, inhibit secretion function, and exhibit astringent and antidiarrheal effects. It promotes digestion and growth. Pine needle powder extract, extracted from pine needles, is rich in flavonoids, various vitamins, amino acids, and minerals, and has strong antioxidant effects. Dihydromyricetin's main active ingredient is a flavonoid compound, which has a variety of unique effects such as scavenging free radicals, antioxidation, antithrombosis, antitumor, and anti-inflammation. Dihydromyricetin is a relatively special flavonoid compound, and compared with ordinary flavonoid compounds, it can show better synergistic effects with black soldier fly larvae intestinal extract, poplar flower extract, and pine needle powder extract.

[0024] 2. When black soldier fly larvae intestinal extract and poplar flower extract are combined, the content of organic acids is further increased, which can further improve the resistance of piglets' intestines to pathogens. Moreover, it does not cause obvious stress response in piglets' intestines. During feeding, the amount added is small and the effect is rapid and obvious.

[0025] 3. This case uses a microemulsion formulation as an oral preparation, which can be well dispersed in drinking water and act quickly on the intestines. It contains compound vitamins and amino acids, among which threonine, arginine and glutamic acid have significant promoting effects on inhibiting intestinal stress response and improving intestinal motility.

[0026] The above improvements can significantly enhance the intestinal health of young livestock and poultry, with low dosage and rapid results. Detailed Implementation

[0027] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0028] Example 1

[0029] Preparation method of microemulsion:

[0030] Step 1: Mix poplar flower extract, dihydromyricetin, pine needle powder extract, and water, and dissolve them completely to obtain an aqueous phase;

[0031] Step 2: Dissolve the black soldier fly larvae intestinal extract in soybean oil to obtain the oil phase;

[0032] Step 3: Add the emulsifier to the oil phase and disperse it evenly;

[0033] Step 4: Slowly add the aqueous phase to the system in Step 3, and then use a shear emulsifier to emulsify at high speed to obtain an oil-in-water emulsion.

[0034] The formula for each substance is as follows:

[0035] Black soldier fly larvae intestinal extract (refer to Example 1 of 201910055164.7, the contents of glyceryl oleate, lauric acid and myristic acid in the effective components are 18.02%, 7.71% and 15.51% respectively) 1g;

[0036] Poplar flower extract (Baoji Fangcheng Biotechnology Development Co., Ltd., specification 20:1, brownish-yellow powder) 2g;

[0037] Dihydromyricetin (powder, net content 98%) 0.25g;

[0038] Pine needle powder extract (Xi'an Youshuo Biotechnology Co., Ltd., specification 10:1, brownish-yellow powder) 2g;

[0039] 6g of emulsifier;

[0040] 12g soybean oil;

[0041] Remaining water.

[0042] The amounts of each component in the emulsifier are as follows:

[0043] Soybean lecithin 0.6g;

[0044] 1.2g of glyceryl monolaurate;

[0045] Tween 80 4.2g.

[0046] Example 2

[0047] Preparation method of microemulsion:

[0048] Step 1: Mix poplar flower extract, dihydromyricetin, and pine needle powder extract with water until fully dissolved to obtain an aqueous phase;

[0049] Step 2: Dissolve the black soldier fly larvae intestinal extract in soybean oil to obtain the oil phase;

[0050] Step 3: Add the emulsifier to the oil phase and disperse it evenly;

[0051] Step 4: Slowly add the aqueous phase to the system in Step 3, and then use a shear emulsifier to emulsify at high speed to obtain an oil-in-water emulsion.

[0052] The formula for each substance is as follows:

[0053] Black soldier fly larvae intestinal extract (refer to Example 1 of 201910055164.7, the contents of glyceryl oleate, lauric acid and myristic acid in the effective components are 18.02%, 7.71% and 15.51% respectively) 1g;

[0054] Poplar flower extract (Baoji Fangcheng Biotechnology Development Co., Ltd., specification 20:1, brownish-yellow powder) 3g;

[0055] Dihydromyricetin (powder, net content 98%) 0.15g;

[0056] Pine needle powder extract (Xi'an Youshuo Biotechnology Co., Ltd., specification 10:1, brownish-yellow powder) 3g;

[0057] 8g of emulsifier;

[0058] 10g soybean oil;

[0059] Remaining water.

[0060] The amounts of each component in the emulsifier are as follows:

[0061] Soybean lecithin 0.8g;

[0062] 1.6g of glyceryl monolaurate;

[0063] Tween 80 5.6g.

[0064] Example 3

[0065] Preparation method of microemulsion:

[0066] Step 1: Mix poplar flower extract, dihydromyricetin, pine needle powder extract, and water, and dissolve them completely to obtain an aqueous phase;

[0067] Step 2: Dissolve the black soldier fly larvae intestinal extract in soybean oil to obtain the oil phase;

[0068] Step 3: Add the emulsifier to the oil phase and disperse it evenly;

[0069] Step 4: Slowly add the aqueous phase to the system in Step 3, and then use a shear emulsifier to emulsify at high speed to obtain an oil-in-water emulsion.

[0070] The formula for each substance is as follows:

[0071] Black soldier fly larvae intestinal extract (refer to Example 1 of 201910055164.7, the contents of glyceryl oleate, lauric acid and myristic acid in the effective components are 18.02%, 7.71% and 15.51% respectively) 1g;

[0072] Poplar flower extract (Baoji Fangcheng Biotechnology Development Co., Ltd., specification 20:1, brownish-yellow powder) 1g;

[0073] Dihydromyricetin (powder, net content 98%) 0.35g;

[0074] Pine needle powder extract (Xi'an Youshuo Biotechnology Co., Ltd., specification 10:1, brownish-yellow powder) 1g;

[0075] 5g of emulsifier;

[0076] 14g soybean oil;

[0077] Remaining water.

[0078] The amounts of each component in the emulsifier are as follows:

[0079] Soybean lecithin 0.5g;

[0080] 1g of glyceryl monolaurate;

[0081] Tween 80 3.5g.

[0082] Example 4

[0083] Preparation method of microemulsion:

[0084] Step 1: Mix poplar flower extract, dihydromyricetin, pine needle powder extract, water, water-soluble vitamins, and amino acids, and dissolve them thoroughly to obtain the aqueous phase;

[0085] Step 2: Dissolve the black soldier fly larvae intestinal extract and oil-soluble vitamins in soybean oil to obtain the oil phase;

[0086] Step 3: Add the emulsifier to the oil phase and disperse it evenly;

[0087] Step 4: Slowly add the aqueous phase to the system in Step 3, and then use a shear emulsifier to emulsify at high speed to obtain an oil-in-water emulsion.

[0088] The formula for each substance is as follows:

[0089] Black soldier fly larvae intestinal extract (refer to Example 1 of 201910055164.7, the contents of glyceryl oleate, lauric acid and myristic acid in the effective components are 18.02%, 7.71% and 15.51% respectively) 1g;

[0090] Poplar flower extract (Baoji Fangcheng Biotechnology Development Co., Ltd., specification 20:1, brownish-yellow powder) 2g;

[0091] Dihydromyricetin (powder, net content 98%) 0.3g;

[0092] Pine needle powder extract (Xi'an Youshuo Biotechnology Co., Ltd., specification 10:1, brownish-yellow powder) 2g;

[0093] 0.5g of water-soluble vitamins;

[0094] 0.5g of oil-soluble vitamins;

[0095] 1g of amino acid composition;

[0096] 6g of emulsifier;

[0097] 12g soybean oil;

[0098] Remaining water.

[0099] The amounts of each component in the emulsifier are as follows:

[0100] Soybean lecithin 0.6g;

[0101] 1.2g of glyceryl monolaurate;

[0102] Tween 80 4.2g;

[0103] The ratio of oil-soluble vitamins is: Vitamin A: Vitamin D: Vitamin E: Vitamin K = 1:1:3:0.1.

[0104] The ratio of water-soluble vitamins is: B vitamins: vitamin C = 1:5.

[0105] The amino acid composition is a complex amino acid provided by Hubei Boyuan Biotechnology Co., Ltd., which contains threonine, arginine and glutamic acid.

[0106] Comparative Example 1

[0107] Similar to Example 1, the formulation of each substance is as follows:

[0108] Dihydromyricetin (powder, net content 98%) 0.25g.

[0109] Poplar flower extract (Baoji Fangcheng Biotechnology Development Co., Ltd., specification 20:1, brownish-yellow powder) 3g;

[0110] Pine needle powder extract (Xi'an Youshuo Biotechnology Co., Ltd., specification 10:1, brownish-yellow powder) 3g;

[0111] 6g of emulsifier;

[0112] 12g soybean oil;

[0113] Remaining water.

[0114] Comparative Example 2

[0115] Similar to Example 1, the formulation of each substance is as follows:

[0116] Black soldier fly larvae intestinal extract (refer to Example 1 of 201910055164.7, the contents of glyceryl oleate, lauric acid and myristic acid in the effective components are 18.02%, 7.71% and 15.51% respectively) 3g;

[0117] Dihydromyricetin (powder, net content 98%) 0.25g.

[0118] 6g of emulsifier;

[0119] 12g soybean oil;

[0120] Remaining water.

[0121] Application performance testing

[0122] Establishment of a pathological model of diarrhea in weaned piglets induced by artificial infection with K88

[0123] I. Evaluation of Prevention Effectiveness

[0124] 1) Feeding management and grouping of experimental piglets

[0125] 216 healthy three-way crossbred (Duroc × Landrace × Large White) weaned piglets with similar initial weights at 28 days were selected and randomly divided into 6 groups according to sex and weight (healthy control group, infection control group, group I (Example 1), group II (Example 4), group III (Comparative Example 1), and group IV (Comparative Example 2), with 3 replicates in each group and 12 weaned piglets (half male and half female) in each replicate).

[0126] All groups were fed a corn-soybean meal basal diet. The pre-trial period was 7 days, and the experimental period was 7 days. During the pre-trial period, piglets found to have diarrhea were culled. The basal diets were all formulated according to NRC (2012) standards.

[0127] All experimental piglets were raised on raised beds in a closed weaning piglet house with natural lighting and free access to feed and water. Experimental groups are shown in Table 1.

[0128] Table 1 Experimental Grouping and Design

[0129]

[0130]

[0131] Remark:

[0132] No other nutrients were added to the drinking water of the healthy control group and the infected control group;

[0133] Add 10 kg / ton of the microemulsion from Example 1 to the drinking water of Group I;

[0134] Add 10 kg / ton of the microemulsion from Example 4 to the drinking water of Group II;

[0135] Add 10 kg / ton of the microemulsion from Comparative Example 1 to the drinking water of Group III;

[0136] Add 10 kg / ton of microemulsion from Comparative Example 2 to the drinking water of Group IV.

[0137] The microemulsion was added to the drinking water according to the above protocol for 7 days during the experimental period.

[0138] 2) Artificial infection experiment

[0139] At the end of the pre-test period, weaned piglets in the control group, group I, group II, and group III were orally administered 2 ml / head of sterilized 1.2% sodium bicarbonate half an hour before viral challenge. Half an hour later, artificial infection and viral challenge were performed, with each piglet orally administered 2 ml / head of E. coli K88 bacterial solution (LD50). 50 5×10 9 (CFU / mL)

[0140] 3) Administration method

[0141] Five days prior to viral challenge, piglets were administered medication as per Table 1 for five consecutive days. Virus challenge commenced on day 5, followed by two days of continued medication. Neither the infected nor healthy control groups received medication. The observation period was seven days, with free access to feed, and diarrhea was recorded in piglets. Piglets that survived seven days were disposed of harmlessly.

[0142] 4) Detection of Escherichia coli count in feces

[0143] The differential culture medium method was used to detect the number of Escherichia coli in feces. Accurately collect 10.0 g of feces and count the colonies using MacConkey medium according to the standard colony counting method. The colony count is expressed as lg (CFU / g).

[0144] 5) Efficacy evaluation indicators

[0145] a. Piglet mortality rate, cure rate, and diarrhea rate; b. Total E. coli count in feces.

[0146] 6) Data Analysis

[0147] All experimental data were statistically analyzed using SPSS 13.0. One-way ANOVA and Duncan's multiple comparisons were performed. Results are presented as mean ± standard error (M ± SE). Mortality and diarrhea rates were expressed as χ². 2 Test and analysis.

[0148] 7) Test Results

[0149] As shown in Tables 2 and 3, the microemulsion groups of the composition can reduce the negative impact on the growth performance of piglets after infection to varying degrees, with obvious preventive effects. They can significantly reduce mortality and diarrhea rate (P<0.01) and significantly reduce the number of Escherichia coli in feces (P<0.01).

[0150] The experimental results can be found in Table 2.

[0151] Table 2. Weight changes of weaned puppies before and after onset of disease in different treatment groups (unit: kg)

[0152]

[0153] Note: Data with the same or no subtitle in the same column indicate no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01). The same applies below.

[0154] Table 3. Records of diarrhea in weaned piglets in different treatment groups

[0155]

[0156] 2. Artificial infection of chickens with bacteria O 78 Trial on the prevention and control efficacy of Salmonella enteritidis

[0157] (1) Feeding management and grouping of experimental chickens

[0158] The experiment was conducted in September 2021 at the Baiyun Experimental Base of the Institute of Animal Science, Guangdong Academy of Agricultural Sciences. 720 healthy fast-growing yellow-feathered broilers (1 day old) were randomly selected and raised to 14 days old. Chickens with similar initial weights (P>0.05) were randomly divided into 6 groups (healthy control group, group I, group II, group III, group IV, and infection control group), with 6 replicates per group and 20 chickens per replicate. Both the healthy and infection control groups were fed a corn-soybean meal basal diet (formulated according to NRC (2012) chicken nutritional requirements). Artificial infection trials began on day 9 of age for each group. All experimental chickens were floor-raised in enclosed broiler houses with sawdust as bedding, 24-hour lighting, and free access to feed and water. The experiment employed a routine immunization program and routine disinfection. The basal diet consisted of corn and soybean meal, selected according to NRC (1994) and combined with the "Chicken Feeding Standards" (NY / T33-2004).

[0159] (2) Artificial infection and treatment

[0160] a. Artificial infection with bacteria O 78 Experiment: Ninety chickens were selected from each of the following groups: Group I, Group II, Group III, Group IV, and the infection control group. Each chicken was subcutaneously injected with 0.5 ml (LD50) of bacterial suspension. 50 Approximately 5.7 × 10 8(cfu / ml). After challenge, chickens were administered 0.5 mL of microemulsion (0.01 g / mL) from groups I, II, III, and IV orally for 5 consecutive days. All groups were fed a corn-soybean meal basal diet for one week with free access to feed, and mortality was recorded. Chickens that survived one week were disposed of harmlessly.

[0161] b. Artificial infection with Salmonella enteritidis: 90 chickens were selected from each of groups I, II, III, IV, and the infection control group. Each chicken was intramuscularly injected with 0.5 ml of Salmonella enteritidis suspension (LD50 approximately 4.8 × 10⁻⁶). 8 After challenge with the virus, chickens were administered 0.5 mL of microemulsion (0.01 g / mL) from groups I, II, III, and IV orally for 5 consecutive days. All groups were fed a corn-soybean meal basal diet and observed for one week with free access to feed. Chicken mortality was recorded. Chickens that did not die after one week were disposed of harmlessly.

[0162] (3) Evaluation indicators of prevention and control effectiveness

[0163] a. Detection of bacteria and Salmonella content in feces: Accurately collect 10.0g of feces and count the colonies using MacConkey medium and Salmonella chromogenic medium according to the conventional colony counting method. The colony count is expressed as lg (CFU / g).

[0164] b. Chicken mortality rate, cure rate, and diarrhea rate;

[0165] c. Total number of bacteria in feces, including total number of Escherichia coli and Salmonella in feces.

[0166] 4) Data Analysis

[0167] All experimental data were statistically analyzed using SPSS 13.0. One-way ANOVA and Duncan's multiple comparisons were performed. Results are presented as mean ± standard error (M ± SE). Mortality, cure rate, and diarrhea rate were expressed as χ². 2 Test analysis. The same applies below.

[0168] 5) Test Results

[0169] Table 4 shows that the microemulsions of the composition significantly improved the growth performance of artificially infected chickens with bacterial diseases (P<0.01), significantly reduced mortality and increased cure rate (P<0.01), and significantly reduced the bacterial content in feces (P<0.01); the effect was: Group II > Group I > Group IV > Group III. Table 5 shows that the microemulsions of the composition significantly improved the growth performance of artificially infected chickens with bacterial diseases (P<0.01), significantly reduced mortality (P<0.01) and diarrhea rate (P<0.01), and significantly reduced the number of bacteria in feces (P<0.01). Therefore, Group II of the composition can effectively prevent and treat chicken bacterial diseases and enteritis-type salmonellosis.

[0170] Table 4 Artificially induced chicken infection with chicken bacteria O 78 Efficacy test with enteritis-type Salmonella

[0171]

[0172] Note: Data with the same or no subtitle in the same column indicate no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

[0173] Table 5 Artificially induced chicken infection with chicken bacteria O 78 Efficacy test with enteritis-type Salmonella

[0174]

[0175]

[0176] Note: Data with the same or no subtitle in the same column indicate no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

[0177] Aquaculture Case 1:

[0178] 1. Feeding and management of experimental animals

[0179] The application effect was evaluated at a large-scale pig farm in Shaoguan, Guangdong. Six weaned piglet pens were selected and randomly divided into a healthy control group (3 pens) and a combination II group (3 pens). Four pens were selected from the control group and the combination II group as experimental groups, with 50 weaned piglets (7.84±0.65kg three-way crossbred (Duroc × Landrace × Large White) weaned piglets) in each pen. The healthy control group was fed a corn-soybean meal basal diet, while the combination II group was fed a corn-soybean meal basal diet plus 1% of combination II (prepared according to Example 4). The experimental period was 14 days. The basal diet was prepared according to the NRC (2012) standard. All piglets were housed in temperature-controlled, closed pens. During the experiment, all piglets had free access to feed and water. At the end of the experiment, the weaned piglets were weighed and the feed was collected. ADG, ADFI, and F:G were calculated for each replicate group. Throughout the trial, the feces or anal condition of each piglet were recorded at 8:30 a.m. every day, and the diarrhea rate was recorded.

[0180] 2. Sample collection and testing of experimental animals

[0181] One day before the end of the experiment (14 days), feed was removed but water was not. Five weaned piglets were randomly selected from each replicate group for weighing, blood collection, and slaughter for intestinal extraction. Indicator detection:

[0182] (1) Serum cytokine detection: Serum was collected by centrifugation of whole blood. The levels of pro-inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-a (TNF-a), as well as anti-inflammatory factors interferon-γ (IFN-γ) and interleukin-10 (IL-10) in the serum were detected using a detection kit. All kits were purchased from Shanghai Enzyme-Linked Biotechnology Co., Ltd.

[0183] (2) Detection of Escherichia coli count in fecal waste: The differential culture medium method was used to detect the number of Escherichia coli in feces. Accurately collect 10.0g of feces and count the colonies using MacConkey medium according to the conventional colony counting method. The colony count is expressed as lg (CFU / g).

[0184] (3) Detection of antioxidant and mucosal immune function indicators in the jejunum of piglets:

[0185] Antioxidant indicators: The levels of glutathione peroxidase (GSH-Px), superoxide dismutase (T-SOD), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) were all determined using reagent kits. All reagent kits were purchased from Nanjing Jiancheng Bioengineering Institute.

[0186] Mucosal immune function indicators: The levels of immunoglobulin G (IgG), secretory immunoglobulin A (sIgA), tumor necrosis factor-α (TNF-α), and the anti-inflammatory factor interferon-γ (IFN-γ) were all determined using kits. All kits were purchased from Shanghai Enzyme-Linked Biotechnology Co., Ltd.

[0187] All experimental data were statistically analyzed using SPSS 13.0. One-way ANOVA and Duncan's multiple comparisons were performed. Results are presented as mean ± standard error (M ± SE). Diarrhea rate was expressed as χ². 2 Test and analysis.

[0188] The results, as shown in Tables 6 to 8, indicate that Composition II significantly improved the growth performance of weaned piglets, significantly reduced piglet diarrhea, and effectively inhibited the number of harmful bacteria; it also increased the content of anti-inflammatory factors in piglet serum and significantly reduced the release of pro-inflammatory factors. Furthermore, it significantly improved the antioxidant capacity and immunity of the piglet jejunum, thereby improving intestinal health.

[0189] Table 6. Effects of Composition II on growth performance, diarrhea rate, and number of pathogenic bacteria in piglets 14 days post-weaning.

[0190]

[0191] Note: Data in the same row with the same letter or no letter above the label indicates no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01); the same applies below.

[0192] Table 7. Effects of Composition I on serum cytokines in piglets 14 days post-weaning.

[0193]

[0194]

[0195] Note: Data in the same row with the same letter or no letter above the label indicates no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

[0196] Table 8. Determination of antioxidant capacity and immune function indicators in the jejunum of weaned piglets.

[0197]

[0198] Note: Data in the same row with the same letter or no letter above the label indicates no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

[0199] Aquaculture Case 2:

[0200] 1. Feeding and management of experimental animals

[0201] The application effect was evaluated in a large-scale chicken farm in Yingde City, Qingyuan. Eight broiler houses were selected and randomly divided into a healthy control group (4 houses) and a composition II group (4 houses). Six pens were selected from the control group and the composition II group as experimental subjects, with 500 chickens in each pen. The healthy control group was fed a corn-soybean meal basal diet with free access to water. The composition II group was fed a corn-soybean meal basal diet with continuous addition of composition II microemulsion (prepared according to Example 4) to the drinking water, with free access to water. The experimental period for each group was 42 days. The basal diet was formulated according to NRC (1994) and combined with the People's Republic of China Agricultural Industry Standard "Chicken Feeding Standard" (NY / T33-2004). All chicks were raised in temperature-controlled, closed houses. During the experiment, all broilers had free access to feed and water. At the end of the experiment, various growth performance indicators of the broilers were measured. Throughout the experiment, the feces of each chicken or the anal condition of piglets were recorded at 8:30 am every day, and the diarrhea rate was recorded.

[0202] 2. Sample collection and testing of experimental animals

[0203] One day before the end of the experiment (42 days), feed was removed but water was not. Fifty birds were randomly selected from each pen in each group for weighing, blood collection, and ileum mucosa retrieval. Various indicators were measured:

[0204] (1) Serum cytokine detection: Serum was collected by centrifuging whole blood from chickens. The levels of pro-inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-a (TNF-a), as well as anti-inflammatory factors interferon-γ (IFN-γ) and interleukin-10 (IL-10) in the serum were detected using a detection kit. All kits were purchased from Shanghai Enzyme-Linked Biotechnology Co., Ltd.

[0205] (2) Detection of antioxidant and mucosal immune function indicators in the ileum of broilers:

[0206] Antioxidant indicators: The levels of glutathione peroxidase (GSH-Px), superoxide dismutase (T-SOD), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) were all determined using reagent kits. All reagent kits were purchased from Nanjing Jiancheng Bioengineering Institute.

[0207] Mucosal immune function indicators: The levels of immunoglobulin G (IgG), secretory immunoglobulin A (sIgA), tumor necrosis factor-α (TNF-α), and the anti-inflammatory factor interferon-γ (IFN-γ) were all determined using kits. All kits were purchased from Shanghai Enzyme-Linked Biotechnology Co., Ltd.

[0208] (3) Detection of Escherichia coli count in fecal waste: The differential culture medium method was used to detect the number of Escherichia coli in feces. Accurately collect 10.0g of feces and count the colonies using MacConkey medium according to the conventional colony counting method. The colony count is expressed as lg (CFU / g).

[0209] All experimental data were statistically analyzed using SPSS 13.0. One-way ANOVA and Duncan's multiple comparisons were performed. Results are presented as mean ± standard error (M ± SE). Diarrhea rate was expressed as χ². 2 Test and analysis.

[0210] The results showed that, as shown in Tables 9 to 11, Composition II significantly improved the growth performance of 42-day-old broilers, significantly reduced broiler diarrhea, effectively inhibited the number of harmful bacteria, significantly increased the content of anti-inflammatory factors in broiler serum and significantly reduced the release of pro-inflammatory factors in serum, and significantly improved the antioxidant capacity and immunity of chicken ileum, thereby improving intestinal health.

[0211] Table 9. Effects of Composition II on growth performance, diarrhea rate, and number of pathogenic bacteria in broilers aged 1-42 days.

[0212]

[0213] Note: Data in the same row with the same letter or no letter above the label indicates no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

[0214] Table 10 Effects of Composition II on serum cytokines in 42-day-old broilers

[0215]

[0216] Note: Data in the same row with the same letter or no letter above the label indicates no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

[0217] Table 11 Determination of antioxidant capacity and immune function indicators in the ileum of broilers

[0218]

[0219]

[0220] Note: Data in the same row with the same letter or no letter above the label indicates no significant difference (P>0.05), different lowercase letters indicate significant difference (P<0.05), and different uppercase letters indicate extremely significant difference (P<0.01).

[0221] in conclusion:

[0222] The following conclusions can be drawn from the above prevention and treatment experiments:

[0223] 1. Black soldier fly larvae intestinal extract has excellent effects in treating and preventing diarrhea in piglets.

[0224] 2. The combination of black soldier fly larvae intestinal extract and poplar flower extract, dihydromyricetin, and pine needle powder extract is superior to the combination of poplar flower extract, pine needle powder extract, dihydromyricetin, and black soldier fly larvae intestinal extract and dihydromyricetin.

[0225] When the composition was used in infection models of piglets and chickens, it was found that its inhibition of pathogenic bacteria was particularly significant when the preferred embodiment (Example 4) was adopted.

Claims

1. A composition for regulating the intestinal health of livestock and poultry, comprising an intestinal extract of black soldier fly larvae, characterized in that, It also includes poplar flower extract, dihydromyricetin, and pine needle powder extract; the weight ratio of the black soldier fly larvae intestinal extract, poplar flower extract, dihydromyricetin, and pine needle powder extract is 1:0.5-3:0.1-0.3:0.5-3; the black soldier fly larvae intestinal extract contains 18.0wt% to 22.0wt% oleic acid glycerides, 7.7wt% to 10.0wt% lauric acid, and 15.5wt% to 18.0wt% myristic acid.

2. The composition for regulating the intestinal health of livestock and poultry according to claim 1, characterized in that, The weight ratio of the black soldier fly larvae intestinal extract, poplar flower extract, dihydromyricetin, and pine needle powder extract is 1:1-2:0.2-0.3:1-2.

3. A microemulsion beneficial to the intestinal health of livestock and poultry, characterized in that, Includes an oil phase, an aqueous phase, the composition as described in any one of claims 1-2, and an emulsifier; The emulsifier constitutes 5-8% of the total weight of the microemulsion; The composition comprises 1-5% of the total weight of the microemulsion; The oil phase comprises 10-15% of the total weight of the microemulsion.

4. The microemulsion beneficial to the intestinal health of livestock and poultry according to claim 3, characterized in that, The emulsifier is composed of soybean lecithin, glyceryl monolaurate and Tween 80 in a weight ratio of 1:2:

7.

5. The microemulsion beneficial to the intestinal health of livestock and poultry according to claim 3, characterized in that, It also includes oil-soluble vitamins, water-soluble vitamins, and amino acid compositions; The amount of each of the oil-soluble vitamins, water-soluble vitamins, and amino acid composition shall not exceed 1% of the total weight of the microemulsion.

6. The microemulsion beneficial to the intestinal health of livestock and poultry according to claim 5, characterized in that, The oil-soluble vitamins include vitamin A, vitamin D, vitamin E, and vitamin K; the weight ratio of vitamin A, vitamin D, vitamin E, and vitamin K is 1:0.1-10:0.1-10:0-0.

1. The water-soluble vitamins include B vitamins and vitamin C; the ratio of B vitamins to vitamin C is 1:2-5.

7. A method for preparing a microemulsion as described in any one of claims 3-6, characterized in that, The oil-soluble component is added to the oil phase, the water-soluble component is added to the aqueous phase, the oil phase and the emulsifier are then mixed uniformly, and finally the aqueous phase is added. The mixture is then sheared and dispersed to obtain an oil-in-water microemulsion.

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