CN107114562B

CN107114562B - Anti-inflammatory and antibacterial and antioxidant composite plant water extract and preparation method and application thereof

Info

Publication number: CN107114562B
Application number: CN201710271907.5A
Authority: CN
Inventors: 陶新; 徐子伟; 吴宛澐; 孙雨晴; 邓波; 门小明; 吴杰; 周文俊
Original assignee: Zhejiang Academy of Agricultural Sciences
Current assignee: Zhejiang Academy of Agricultural Sciences
Priority date: 2017-04-24
Filing date: 2017-04-24
Publication date: 2020-08-14
Anticipated expiration: 2037-04-24
Also published as: CN107114562A

Abstract

The invention relates to a composite plant water extract and application thereof in the field of feed and animal health products, in particular to an anti-inflammatory and anti-oxidation composite plant water extract and a preparation method and application thereof. The anti-inflammatory and antibacterial composite plant water extract is prepared by extracting the following plant raw materials in percentage by mass: pomegranate bark 4.17%, lemon 54.16%, cassia twig 4.17%, sweet wormwood 16.67% and magnolia bark 20.83%. The invention discloses a set of simple process suitable for extracting multiple active ingredients of plants, and is suitable for processing and production of various small and medium-sized enterprises. The plant raw materials for forming the composite plant extract in the invention are all low-price or medium-price, and have abundant domestic resources and high cost performance. Compared with the original formula, the pomegranate rind with higher cost performance is kept, the cost of the compound formula is greatly reduced, and the application dosage on piglets can be obviously reduced.

Description

Anti-inflammatory and antibacterial and antioxidant composite plant water extract and preparation method and application thereof

Technical Field

The invention relates to a composite plant water extract and application thereof in the field of feed and animal health products, in particular to an anti-inflammatory and anti-oxidation composite plant water extract and a preparation method and application thereof.

Background

Feeding antibiotics have contributed to the development of livestock and poultry breeding industry, but the problems of drug resistance, drug residues and the like are increasingly highlighted. After antibiotics are forbidden, the research of antibiotic substitutes becomes a problem to be solved for the first time in the breeding industry. The plant extract is rich in various active ingredients such as protein, glucoside and phenolic acid, has the functions of bacteriostasis, antioxidation, anti-stress, antivirus, immunoregulation and the like, is not easy to generate the problems of bacterial drug resistance, drug residue and the like, and becomes a research hotspot of antibiotic substitutes. However, most of the currently studied extracts are extracted from a single plant raw material, so that the problems of low content of bioactive components, single function, insignificant effect and the like exist. For the compound extract, the defects of weak compound scientificity, high addition amount, high cost and the like exist. Therefore, if a compound plant extract with multiple functions and high cost performance can be screened out to replace the antibiotics for feeding piglets, a great problem of the livestock breeding industry can be undoubtedly solved. The compound plant extract developed in the early stage by the applicant has good effects of promoting growth and replacing feed antibiotics on weaned pigs, but has limited application prospect due to the problems of high price, large addition amount and the like.

Disclosure of Invention

The invention aims to provide a composite plant water extract which is used for replacing feed antibiotics and finally achieves the 'environment-friendly, non-toxic, safe and green' breeding goal. The second purpose of the invention is to provide the preparation method of the composite plant aqueous extract, which has simple process and low cost. The third purpose of the invention is to provide the application of the composite plant water extract.

In order to achieve the first object, the invention adopts the following technical scheme:

the anti-inflammatory and antibacterial composite plant water extract is prepared by extracting the following plant raw materials in percentage by mass:

pomegranate rind 4.17%

54.16 percent of lemon

4.17 percent of cassia twig

16.67 percent of sweet wormwood herb

20.83 percent of officinal magnolia.

In order to achieve the second object, the invention adopts the following technical scheme:

the method for preparing the composite plant aqueous extract comprises the following steps:

1) grinding and crushing plant raw materials, and mixing, wherein the ratio of material to water is 1: 20, placing the mixture into an extraction equipment tank, heating to 80 ℃, extracting for 6 hours, transferring the mixture into a concentration tank, and concentrating the mixture to the specific gravity of 1.25 under the conditions of 0.085MPa and 60 ℃;

2) transferring the concentrated product to a spray drying device, and spray drying the concentrated product under the conditions of air inlet temperature of 150 ℃, air flow rate of 500L/H and feeding amount of 20%.

In order to achieve the third object, the present invention adopts the following technical solutions:

the composite plant water extract is used for preparing animal feed additives for replacing antibiotics.

The invention also provides an animal feed additive, and the animal feed additive package is the composite plant water extract.

The invention also provides animal feed which comprises the composite plant water extract.

In summary, the technical scheme adopted by the invention is as follows:

(1) in order to realize the primary purpose of the invention, firstly, a set of simple process parameters suitable for extracting a plurality of active ingredients of plants is searched and established: the material-water ratio is 1: 20. extracting at 80 deg.C for 6 hr, concentrating the extractive solution at 60 deg.C to specific gravity of 1.25, and spray drying (air inlet temperature of 150 deg.C, air flow rate of 500L/H, and feed rate of 20%).

(2) Then, the in-vitro anti-inflammatory, bacteriostatic and antioxidant capacities are taken as evaluation indexes, and the raw material composition of the composite plant extract is preliminarily screened from 23 plant raw materials: pomegranate rind, magnolia bark, cassia twig, sweet wormwood herb and lemon. The original formula comprises the following raw materials: pomegranate rind, folium artemisiae argyi, fructus schisandrae and fructus evodiae.

(3) Further, the formula (pomegranate peel 1g, cassia twig 1g, sweet wormwood 4g, magnolia bark 5g and lemon 13 g) of the composite plant extract and the antibacterial function (MIC 125 mg/mL) anti-Inflammatory (IC) of the extract are verified through an orthogonal combination experiment₅₀0.138mg/mL) and antioxidant capacity (IC)₅₀0.40 mg/mL). The original formula (pomegranate rind 1g, argy wormwood leaf 3g, south schisandra fruit)Seed 9g and fructus evodiae 9 g) and verification of bacteriostatic function (MIC is 250 mg/mL) and antioxidant capacity (IC)₅₀6.46mg/mL), its anti-inflammatory activity was not detected.

(4) Respectively and continuously gavage 125, 250, 500 and 1000 mg/kg-bw for the mice^-1The compound extract has good mental state, bright hair color and no abnormal death phenomenon in 21 days, and the addition amount of the compound plant extract is less than 125 mg/kg-bw after comprehensive analysis^-1。

(5) 0.3mg/ml (equivalent to 100 mg/kg. bw) was added to the mouse drinking water^-1Dosage) of the composition is 21 days, the daily gain of mice can be remarkably improved by 13.3 percent (P is 0.008), and the material-to-weight ratio is remarkably reduced by 10.8 percent (P)<0.05). The original formulation was not studied, but only the mice were gavaged with 1000 mg/kg-bw per day^-1The dosage is continuously gavage for 14 days, so that the daily gain of the mice can be increased.

(6) 0.3mg/ml (equivalent to 100 mg/kg. bw) was added to the mouse drinking water^-1Dose) for 21 days, the diarrhea rate of the mice can be reduced after the mice are subjected to LPS modeling treatment, and the villus height of the small intestine and the value (P) of the height of the villus/the depth of crypt are obviously improved<0.05) and remarkably reduces the gene expression (P) of proinflammatory factors IL-1 β and IL-6 of the small intestinal mucosa<0.05）。

(7) Compared with the original formula, the dosage of the compound plant extract in mice is only 10% of that of the original formula, so that the cost performance of the compound plant extract is improved to a great extent, and the compound plant extract has a good application prospect in livestock breeding.

The invention has the following advantages:

1. the invention discloses a set of simple process suitable for extracting multiple active ingredients of plants, and is suitable for processing and production of various small and medium-sized enterprises.

2. The plant raw materials for forming the composite plant extract in the invention are all low-price or medium-price, and have abundant domestic resources and high cost performance.

3. Compared with the original formula, the pomegranate rind with higher cost performance is kept, the cost of the compound formula is greatly reduced, and the application dosage on piglets can be obviously reduced.

Drawings

FIG. 1 is a photograph of a photomicrograph of the villi of the jejunum of a mouse 4 hours after LPS injection. FIG. 1A shows a normal control group (saline injection 4 h), FIG. 1B shows an LPS group (LPS injection 4 h), and FIG. 1C shows an anti-inflammatory complex group (LPS injection 4 h).

FIG. 2 is a photograph of a photomicrograph of the villi of the jejunum of a mouse 24 hours after LPS injection. FIG. 2A is LPS group (LPS injection 24 h), and FIG. 2B is anti-inflammatory complex group (LPS injection 24 h).

FIG. 3 is a graph showing the gene expression levels of proinflammatory factors in the jejunal mucosa of mice 4 hours after LPS injection.

FIG. 4 is a graph showing the gene expression levels of proinflammatory factors in the jejunal mucosa of mice 24 hours after LPS injection.

Detailed Description

Firstly, extracting and screening plant raw materials and detecting compound active ingredients

1. The extract comprises 23 plant materials including herba Artemisiae Annuae, fructus Ligustri Lucidi, fructus Citri Limoniae, herba Euphorbiae Helioscopiae, ramulus Cinnamomi, pericarpium Granati, fructus Citri Limoniae, radix Acanthopanacis Senticosi, Achyranthis radix, herba Gynostemmatis, semen Ginkgo, cortex Fraxini, herba Houttuyniae, fructus Psoraleae, cortex Magnolia officinalis, rhizoma Cyperi, herba Artemisiae Scopariae, herba Rosmarini officinalis, cortex Cinnamomi, Atractylodis rhizoma, herba Epimedii, Glycyrrhrizae radix, folium Bambusae, herba Filipendulae, etc.

2. The extraction process comprises the following steps: grinding and crushing plant raw materials, and mixing the raw materials according to a material-water ratio of 1: 20, placing the mixture into an extraction equipment tank, heating to 80 ℃, extracting for 6 hours, transferring the mixture into a concentration tank, and concentrating the mixture to 1.25 at the temperature of 60 ℃ under the pressure of 0.085 MPa. Transferring the concentrated product to a spray drying device, and spray drying the concentrated product under the conditions of air inlet temperature of 150 ℃, air flow rate of 500L/H and feeding amount of 20%.

3. Determination of anti-inflammatory rate: the ultraviolet spectrophotometer determines 23 the inhibition rate of the plant water extract on the activity of Lipoxygenase (LOX) to characterize the in vitro anti-inflammatory activity of the plant water extract. The specific operation is as follows: (1) test groups: adding 1 mu L of 0.5g/mL Chinese medicinal material water extract into a 1.5mL eppendorf tube containing 20 mu L of enzyme solution, carrying out water bath at 30 ℃ for 30min, adding 150 mu L of substrate, reacting at 30 ℃ for 3 min, adding 500 mu L of absolute ethyl alcohol to terminate the reaction, and then adding 500 mu L of distilled water to be uniformly mixed to be tested; (2) positive control group: replacing the water extract of the traditional Chinese medicinal materials with 1 mu L of distilled water, and the rest steps are the same as the step (1); (3) negative control group: taking 1 μ L of Chinese medicinal material water extract, replacing enzyme solution with 20 μ L of distilled water, and the rest steps are the same as (1); (4) blank control group: 1 mu L of distilled water is used for replacing the water extract of the traditional Chinese medicinal materials, 500 mu L of absolute ethyl alcohol is added before the substrate is added to stop the reaction, and the rest steps are the same as the step (1). And (3) taking 200 mu L of the solution to be detected to a 96-hole ultraviolet detection plate, and selecting the absorbance OD value of the reaction solution on a microplate reader at the wavelength of 234 nm. The inhibitory activity of the aqueous extract of Chinese medicinal materials is expressed by the anti-inflammatory rate, and the formula is as follows:

ODa, ODb, ODc, ODd represent OD values of the test group, positive control group, negative control group, and blank control group, respectively.

TABLE 1 inhibitory Activity of aqueous plant extracts on lipoxygenase (anti-inflammatory Rate)

Species of extract	Ramulus Cinnamomi	Rhizoma Cyperi	Herba Artemisiae Scopariae	Glossy privet fruit	Herba Epimedii	Cortex Cinnamomi
							Anti-inflammatory Rate (%)	68.06	12.0	72.01	20.0	72.3	85.66
Species of extract	Herba Euphorbiae Helioscopiae	Gynostemma pentaphylla	Root of bidentate achyranthes	Cortex Magnoliae officinalis	White atractylodes rhizome	Licorice root, radix Glycyrrhizae
							Anti-inflammatory Rate (%)	89.34	32.78	6.71	91.25	5.88	1.13
Species of extract	Ginkgo leaf	Cortex Fraxini	Filipendula ulmaria (L.) Kuntze	Rosemary	Pomegranate rind	Radix Et caulis Acanthopanacis Senticosi
							Anti-inflammatory Rate (%)	23.5	47.4	88.92	90.84	96.06	34.29
Species of extract	Sweet wormwood herb	Fructus psoraleae	Lemon	Bamboo leaf	Herba Houttuyniae
							Anti-inflammatory Rate (%)	—	—	91.47	—	—

Note: no anti-inflammatory effect

Through the above studies (table 1), the plant materials with strong anti-inflammatory ability were selected as follows: pomegranate rind, lemon and magnolia bark.

4. And (3) determining the bacteriostatic rate: taking Escherichia coli C83912 as a tested strain, wherein the crude drug concentration of the extract is 0.5g/mL, and the ratio of the crude drug concentration to the crude drug concentration is 1:10 to each medium, inoculating a bacterial solution of 1:1000, culturing on an enzyme label plate at 37 ℃ and 210r/min for 24h, and measuring the OD value at 600 nm. The formula for calculating the bacteriostasis rate is as follows:

TABLE 2 bacteriostatic ratio of aqueous plant extract to E.coli

Species of extract

Ramulus Cinnamomi

Rhizoma Cyperi

Herba Artemisiae Scopariae

Glossy privet fruit

Herba Epimedii

Cortex Cinnamomi

Rate of inhibition of bacteria

+++

—

+++

Species of extract

Herba Euphorbiae Helioscopiae

Gynostemma pentaphylla

Root of bidentate achyranthes

Cortex Magnoliae officinalis

White atractylodes rhizome

Licorice root, radix Glycyrrhizae

Rate of inhibition of bacteria

—

+

—

++

Species of extract

Ginkgo leaf

Cortex Fraxini

Filipendula ulmaria (L.) Kuntze

Rosemary

Pomegranate rind

Radix Et caulis Acanthopanacis Senticosi

Rate of inhibition of bacteria

++

—

+

—

+++

+

Species of extract

Sweet wormwood herb

Fructus psoraleae

Lemon

Bamboo leaf

Herba Houttuyniae

Rate of inhibition of bacteria

+++

—

Note: the bacteria inhibiting rate is 50%, the bacteria inhibiting rate is 30-50%, the bacteria inhibiting rate is 30%, and no bacteria inhibiting effect

Through the above research (table 2), the plant materials with strong bacteriostatic rate are screened out as follows: pomegranate rind, cassia twig and sweet wormwood herb.

5. And (3) measuring the antioxidant capacity: when the crude drug concentration of the extract is detected to be 100, 50, 25, 12.5 and 6.25mg/mL respectively, an enzyme label plate (100 muL/hole) is adopted, and the volume ratio is 1: 1. clearance of DPPH (0.5 mg/mL) at 37 ℃ for 30min, OD at 516nm, according to the formula:

calculating radical clearance and converting the half radical clearance (IC) by using CurveExpert fitting regression equation₅₀）。

TABLE 3 Elimination of DPPH free radicals by aqueous plant extracts IC₅₀Value (antioxidant IC)₅₀）

Species of extract	Ramulus Cinnamomi	Rhizoma Cyperi	Herba Artemisiae Scopariae	Glossy privet fruit	Herba Epimedii	Cortex Cinnamomi
							Oxidation resistant IC₅₀，mg/ml	10.41	36.08	1.04	2.55	5.34	1.5
Species of extract	Herba Euphorbiae Helioscopiae	Gynostemma pentaphylla	Root of bidentate achyranthes	Cortex Magnoliae officinalis	White atractylodes rhizome	Licorice root, radix Glycyrrhizae
							Oxidation resistant IC₅₀，mg/ml	13.08	1.1	199.27	1.54	10.91	78.45
Species of extract	Ginkgo leaf	Cortex Fraxini	Filipendula ulmaria (L.) Kuntze	Rosemary	Pomegranate rind	Radix Et caulis Acanthopanacis Senticosi
							Oxidation resistant IC₅₀，mg/ml	2.87	0.71	8.52	0.31	0.08	50.62
Species of extract	Sweet wormwood herb	Fructus psoraleae	Lemon	Bamboo leaf	Herba Houttuyniae
							Oxidation resistant IC₅₀，mg/ml	2.71	149.17	20.56	25.34	5.19

Through the above studies (table 3), the plant materials with strong antioxidant capacity were selected as follows: pomegranate rind.

5. Orthogonal combinatorial testing: by using L₁₆(5⁴) Experimental design (Table 4), 16 composite plant extracts containing lemon, magnolia officinalis, sweet wormwood, cassia twig and pomegranate bark in different proportions are respectively prepared and tested for in vitro antibacterial rate and antioxidant capacity, and according to test results, the optimal composite extract formula of pomegranate bark 1g, cassia twig 1g, sweet wormwood 4g, magnolia officinalis 5g and lemon 13 g) and the antibacterial function (MIC 125 mg/mL) and anti-Inflammatory (IC) function of the extract formula are verified₅₀0.138mg/mL) and antioxidant capacity (IC)₅₀0.40 mg/mL).

Table 4 orthogonal combination design form of five plant materials

TABLE 5 in vitro bacteriostatic, anti-inflammatory and antioxidant IC of orthogonally designed individual combined plant aqueous extracts₅₀Value of

Combination of	Bacteriostatic ratio (%)	Anti-inflammatory Rate (%)	Oxidation resistant IC₅₀（mg/mL）
				1	34.53	86.93	0.82
2	33.73	78.53	0.82
				3	-4.99	69.55	0.67
4	36.53	75.03	0.41
				5	26.95	57.12	0.29
6	31.74	73.07	0.27
				7	17.76	79.87	0.48
8	38.52	94.31	0.69
				9	59.08	72.21	0.72
10	79.44	91.81	2.01
				11	-1.20	91.67	0.33
12	79.64	92.81	0.26
				13	60.50	73.51	0.29
14	28.54	79.71	0.25
				15	81.04	93.47	1.09
16	71.06	103.91	0.72

According to the orthogonal design and the bacteriostatic, anti-inflammatory and antioxidant results of each combination (table 5), the influence factors of the anti-inflammatory effect are determined by a range analysis method and are as follows in sequence: the combination level of lemon, pomegranate peel, magnolia bark, sweet wormwood and cassia twig, which has the best in-vitro anti-inflammatory effect and has the functions of bacteriostasis and antioxidation is as follows: the compound is marked as an anti-inflammatory compound, wherein the mass of the raw materials corresponding to each level is 1g, 1g, 4g, 5g and 13g respectively, and the pomegranate bark is 1, the cassia twig is 1, the sweet wormwood herb is 1, the mangnolia officinalis is 4 and the lemon is 4. Determining the influence factors of the antioxidant effect by adopting a range analysis method in turn: the combination level of the pomegranate rind, the cassia twig, the magnolia bark, the lemon and the sweet wormwood herb with the best in-vitro antioxidant effect and the functions of bacteriostasis and anti-inflammation is as follows: the compound is marked as an antioxidant compound, wherein the mass of the raw materials corresponding to each level is 7g, 1g, 4g, 3g and 13g respectively, and the weight of the compound is 1, 2 and 4.

6. Preparation of composite plant extract, determination of antibacterial, anti-inflammatory and antioxidant activities

The formula of the composite plant extract comprises the following components:

pomegranate rind 4.17%

54.16 percent of lemon

4.17 percent of cassia twig

16.67 percent of sweet wormwood herb

20.83 percent of officinal magnolia.

The preparation method comprises the following steps: 1) grinding and crushing plant raw materials, and mixing the raw materials according to a material-water ratio of 1: 20, placing the mixture into an extraction equipment tank, heating to 80 ℃, extracting for 6 hours, transferring the mixture into a concentration tank, and concentrating the mixture to the specific gravity of 1.25 under the conditions of 0.085MPa and 60 ℃;

(1) Anti-inflammatory IC₅₀Detection of the value: the aqueous extract was diluted in equal proportions and the anti-inflammatory rate of each concentration gradient was determined as described above. Fitting a regression equation by using CurveExpert software according to the anti-inflammatory rate and the concentration of the aqueous extract, and calculating the concentration of the aqueous extract when the anti-inflammatory rate is 50%, namely anti-inflammatory IC₅₀。

(2) And (3) detection of the bacteriostatic MIC value: the macrobouillon dilution method was used. Concentrating the compound water extract to crude drug concentration of 1g/mL, and diluting 0.5 McLeeb's inoculum with MH liquid culture medium at a ratio of 1: 1000. Taking a sterilized test tube, adding 1mL of sterile broth culture medium, and diluting in multiple times until the concentration of the aqueous extract in each test tube is 500, 250, 125, 62.5 and 31.25 mg/mL respectively; and a positive control and a negative control were prepared simultaneously. All tubes were incubated in a 37 ℃ incubator for 24 h. And after the culture is finished, observing the growth condition of bacteria, wherein the sterile test tube with the lowest concentration is the Minimum Inhibitory Concentration (MIC) of the water extract.

(3) Oxidation resistant IC₅₀Detection of the value: the method is the same as above.

TABLE 6 antibacterial MIC and antiinflammatory IC of anti-inflammatory and antioxidant compounds₅₀And antioxidant IC₅₀Results

As can be seen from Table 6, the anti-inflammatory formulation had a bacteriostatic MIC of 125mg/mL and an anti-inflammatory IC₅₀Value of 0.14, antioxidant IC₅₀The value was 0.40.

7. Application effect of compound plant extracts with different dosages on mice

54 Balb/c female mice were selected and randomly divided into 3 treatments according to body weight: a control group (lavage normal saline), an anti-inflammatory compound group and an antioxidant compound group, and two compoundsAre respectively 125, 250, 500 and 1000 mg/kg · bw ^-14 levels (gavage corresponding aqueous extract), 6 mice per level, 21 days of the experimental period. During the whole feeding test period, all experimental mice have good mental state, bright hair color and no abnormal death phenomenon; as can be seen from tables 7-12, 2 compound high doses of the compound drug have significant effects on the daily gain and the index of part of the viscera (P)<0.05), especially the index of partial organs of antioxidant compound mouse is even 250mg/kg bw^-1The difference of the dosage is still significant with the control group (P)<0.05). Therefore, the growth promoting effect on mice was studied only on the anti-inflammatory complex.

TABLE 7 daily gain and feed intake in mice of different treatment groups

Note: indicates that the significant difference P is less than 0.05

TABLE 8 organ index liver of mice in different treatment groups

TABLE 9 organ index-Kidney of mice in different treatment groups

TABLE 10 organ index of mice in different treatment groups-thymus

TABLE 11 organ index of mice in different treatment groups-spleen

TABLE 12 organ index-Heart of mice in different treatment groups

8. Research on growth promoting effect of anti-inflammatory compound on mice

About 60 Balb/c female mice of about 15g were selected and divided into 2 groups: control group (normal drinking water), anti-inflammatory compound group (drinking water added with 0.3 mg/ml), each group has 3 replicates, and the test period is 21 days. The test results show (table 13): compared with a control group, the anti-inflammatory recovery can remarkably increase the daily gain of mice by 13.3 percent (P is 0.008) and remarkably reduce the material-to-weight ratio by 10.8 percent (P is less than 0.05).

TABLE 13 Effect of anti-inflammatory Compounds on growth Performance in mice

	Control	Anti-inflammatory compound
			Daily food intake g	3.11±0.09	3.25±0.08
Daily gain g	0.15±0.01^a	0.17±0.01^b
			Material to weight ratio	21.21±1.63^a	18.91±1.09^b

9. Anti-inflammatory compound protective effect on intestinal health of mice

45 Balb/c female mice were selected and divided into 3 treatments: the control group (normal drinking water), LPS + compound group (drinking water added with 0.3mg/ml anti-inflammatory compound), feeding test till 21 days, injecting normal saline into the control group, injecting LPS into the other 2 groups, slaughtering and sampling respectively at 4h (8 in each group) and 24h (7 in each group) after LPS injection. As can be seen from FIGS. 1 and 2, the results of the 4h test after LPS injection show (tables 14-16): all mice in LPS group have diarrhea, the diarrhea rate is 100%, and compared with a control group, the CD of the jejunum crypt depth is obviously increased (P < 0.05), and the VH/CD is obviously reduced (P < 0.05); the diarrhea rate of LPS + compound mice is 75%, and compared with LPS group, VH/CD is obviously increased (P is less than 0.05); the gene expression levels of IL-1. beta. and IL-6 in LPS + complex were reduced by 40.1% and 60.8%, respectively (P < 0.01). As can be seen from FIGS. 3 and 4, the test results 24h after LPS injection show that the diarrhea rate of mice in LPS group is 71.43%; compared with LPS group, LPS + compound group has obviously raised jejunum VH (P < 0.05), and IL-1 beta and IL-6 gene expression level lowered 52.27% and 59.46% (P < 0.01), respectively.

TABLE 14 diarrhea in mice of different treatment groups 4h and 24h after LPS injection

TABLE 15 values for jejunal villus height, crypt depth and villus height/crypt depth for different treatment groups of mice 4h post LPS injection

	Control group	LPS group	LPS + compound group
				Fluff height (VH)	474.23±41.25	431.55±42.14	457.79±44.13
Depth of recess (CD)	155.90±21.92^a	225.54±27.05^b	201.04±30.45^b
				Villus height/crypt depth (VH/CD)	3.23±0.28^c	1.95±0.11^a	2.35±0.25^b

TABLE 16 Colon height, crypt depth and villus height/crypt depth of different treatment groups of mice 24h after LPS injection

	LPS group	LPS + anti-inflammatory complex
			Fluff height (VH)	331.00±60.26^a	462.49±38.93^b
Depth of recess (CD)	168.15±15.74^a	219.87±31.75^a
			Villus height/crypt depth (VH/CD)	2.01±0.34^a	2.20±0.30^ab

Claims

1. The anti-inflammatory and antibacterial composite plant water extract is characterized by being prepared by extracting the following plant raw materials in percentage by mass:

pomegranate rind 4.17%

54.16 percent of lemon

4.17 percent of cassia twig

16.67 percent of sweet wormwood herb

20.83 percent of officinal magnolia;

the aqueous extract is prepared by the following method: 1) grinding and crushing plant raw materials, and mixing the raw materials according to a material-water ratio of 1: 20, placing the mixture into an extraction equipment tank, heating to 80 ℃, extracting for 6 hours, transferring the mixture into a concentration tank, and concentrating the mixture to the specific gravity of 1.25 under the conditions of 0.085MPa and 60 ℃;

2. The use of the anti-inflammatory and antioxidant composite plant water extract of claim 1 in the preparation of an animal feed additive for replacing antibiotics.

3. An animal feed additive, which comprises the anti-inflammatory and anti-oxidant composite plant aqueous extract of claim 1.

4. An animal feed, which comprises the anti-inflammatory and anti-oxidant composite aqueous plant extract of claim 1.