CN115887535B - Compound Chinese medicinal residue fermentation liquor for preventing and treating colibacillosis of poultry and preparation method thereof - Google Patents

Abstract

The invention provides a compound Chinese medicinal residue fermentation liquor for preventing and treating colibacillosis of birds and a preparation method thereof, belonging to the technical field of Chinese medicinal residue fermentation liquor. The compound Chinese medicinal dreg fermentation broth is prepared by fermenting lactobacillus rhamnosus PC30-2-1, can reduce the inflammatory cytokine level of pathogenic escherichia coli of artificially infected poultry, improve bursal index, improve appetite and disease resistance of the poultry, promote weight gain, improve production benefit, reduce or replace the use of conventional antibacterial medicines, reduce medicine residues, reduce the selection pressure of the antibacterial medicines, and inhibit or slow down the generation of clinical pathogenic bacteria resistance.

Description

Compound Chinese medicinal residue fermentation liquor for preventing and treating colibacillosis of poultry and preparation method thereof

Technical Field

The invention belongs to the technical field of traditional Chinese medicine residue fermentation liquor, and in particular relates to a compound traditional Chinese medicine residue fermentation liquor for preventing and treating colibacillosis of birds and a preparation method thereof.

Background

Colibacillosis is a common infectious disease of various livestock and poultry, and pathogenic Escherichia coli (E.coli) is an important zoonotic primordium. Not only causes huge economic loss for poultry farming industry and prevents continuous healthy development of the poultry farming industry, but also is extremely easy to cause animal food safety problem and brings potential threat to human health and living environment. At present, the disease is mainly prevented and treated by the antibacterial drugs clinically, but multi-drug resistance, cross-drug resistance and even super-drug resistance strains are continuously generated due to the unreasonable use of the antibacterial drugs, and great difficulty is brought to preventing and treating the disease. For this reason, reducing or eliminating the resistance of pathogenic E.coli is critical for effective control of the disease. The traditional Chinese medicine has various biological activities such as antibiosis, antivirus, immunoregulation and the like, is difficult to generate drug resistance due to various components, and has important significance for effectively preventing and treating bacterial diseases of livestock and poultry caused by pathogenic escherichia coli and guaranteeing human health and ecological environment. With the release of relevant regulations such as national veterinary antibacterial drug use reduction action scheme (2021-2025) of agricultural rural department of China, the use of veterinary traditional Chinese medicines for reducing or replacing conventional veterinary antibacterial drugs has become a new direction for green and healthy development of industry research hotspots and animal husbandry. Lactobacillus rhamnosus is a safe and widely applied probiotic bacterium, can resist the environment of animal digestive tracts, can be planted in human and animal intestinal tracts, and has the effects of regulating intestinal flora, preventing and treating diarrhea, removing toxins, preventing decayed teeth, improving organism immunity and the like. It has the characteristics of acid resistance, oxygen resistance and strong field planting force, and has higher current acceptance.

In conclusion, on the basis of traditional Chinese medicine processing, the lactobacillus fermentation technology is utilized, so that the content of active ingredients of the traditional Chinese medicine can be improved, the active ingredients which are not originally contained in some traditional Chinese medicines can be changed or generated, and the macromolecular substances of the traditional Chinese medicine can be converted into small molecules which can be directly absorbed and utilized by an organism, thereby improving the disease resistance and economic benefit of the animal organism.

Disclosure of Invention

The invention aims to provide a compound traditional Chinese medicine residue fermentation liquor for preventing and treating avian colibacillosis and a preparation method thereof.

The invention firstly provides a preparation method of a compound traditional Chinese medicine residue fermentation liquor for preventing and treating colibacillosis of birds, which comprises the following steps:

step one: lactobacillus rhamnosus PC30-2-1 is obtained by separating from commercial pickle, and strain identification is carried out through 16S rDNA, so as to obtain lactobacillus rhamnosus PC30-2-1 (Lactobacillus rhamnosus PC-2-1) with the preservation number of CCTCC NO: M2022083, wherein the preservation unit is: china center for type culture collection, with the preservation addresses: the preservation date of the university of Chinese Wuhan is: inoculating lactobacillus rhamnosus PC30-2-1 into a culture medium at 37 ℃ for 12-18h under shaking culture at 150r/min at 2022, 1 month and 17 days to obtain seed culture solution;

step two: slicing herba asari, glycyrrhrizae radix, cortex Cinnamomi, radix Angelicae sinensis, radix Paeoniae Rubra, fructus Jujubae and caulis Akebiae, decocting in water, and mixing filtrates;

step three: drying the dregs obtained in the second step, adding distilled water into the obtained dry dregs, sterilizing, and cooling to room temperature to obtain dregs culture solution;

step four: inoculating the seed culture solution in the second step into the residue culture solution in the third step, shake culturing at 37deg.C and 150r/min for 12-18 hr, centrifuging, and collecting fermentation supernatant to obtain compound Chinese medicinal residue fermentation liquor.

Preferably, the culture medium in the first step is MRS liquid culture medium.

Preferably, the second step comprises the following steps: 12-15 parts of asarum, 12-15 parts of liquorice, 12-15 parts of cinnamon, 12-15 parts of angelica sinensis, 12-15 parts of red paeony root, 12-15 parts of Chinese date and 8-10 parts of akebia stem.

Preferably, the second step comprises the following steps: 12 parts of asarum, 12 parts of liquorice, 12 parts of cinnamon, 12 parts of angelica sinensis, 12 parts of red paeony root, 12 parts of Chinese date and 8 parts of akebia stem. Preferably, the second time is 2 times, the first time is 1h, and the second time is 40-45min.

Preferably, the drying temperature in the third step is room temperature, and the drying time is 48 hours.

Preferably, the feed liquid ratio of the Chinese medicine residues to distilled water in the third step is 1:10.

Preferably, the sterilization temperature in the third step is 121 ℃ and the pressure is 103.4kPa.

Preferably, the step four centrifugation and collection includes: centrifuging the residue fermentation broth at 4deg.C at 8000r/min for 10min, vacuum filtering the supernatant with 0.45 μm filter membrane, and concentrating the filtrate by rotary evaporation (45deg.C, 75 r/min) to volume ratio of 1:3.

The invention also provides the compound Chinese medicinal dreg fermentation liquor for preventing and treating the colibacillosis of the poultry, which is obtained by the preparation method.

The pharmacology and the drug property of each raw material medicine in the dregs of decoction are as follows:

asarum herb: warm nature, pungent taste and small toxicity. It enters heart, lung and kidney meridians. Dispelling pathogenic wind, dispelling cold, inducing resuscitation, relieving pain, warming lung, and eliminating phlegm. Belongs to the pungent and warm exterior-releasing herbs of the category of exterior-releasing herbs. 1. Local anesthetic action; 2. inhibiting arthritis; 3. antibacterial effect; 4. has effects in enhancing lipid metabolism and increasing blood sugar, and regulating smooth muscle function. The volatile oil can reduce blood pressure of anesthetized animals, and the decoction can raise blood pressure; 5. anti-inflammatory and anticonvulsant effects; 6. pain relieving, tranquillizing, fever suppressing and fever reducing; 7. antihistaminic and antiallergic responses; 8. exciting respiration.

Licorice root: sweet in taste, mild in nature, and good in effect in invigorating spleen, replenishing qi, clearing away heat and toxic materials, eliminating phlegm, relieving cough, relieving pain, and alleviating drug properties. Is used for treating weakness of spleen and stomach, listlessness, debilitation, palpitation, short breath, cough with excessive phlegm, abdominal pain, limb spasm, acute pain, carbuncle, swelling and sore. Pharmacological actions include: 1. adrenocortical hormone-like action (mineralocorticoid-like action, glucocorticoid-like action); 2. enhancing nonspecific immunity and cellular immunity, but inhibiting humoral immunity; 3. acting on digestive system (antiulcer, liver protecting, spasmolysis); 4. detoxification; 5. anti-inflammatory; 6. cough relieving and phlegm eliminating; 7. an anti-pathogenic microorganism; 8. antiarrhythmic; 9. an anti-tumor; 10. reducing blood lipid and resisting atherosclerosis.

Cinnamon: pungent and sweet in flavor, strong in heat, and enters spleen, kidney, heart and liver meridians, with the effects of tonifying fire and yang, inducing fire to enter sources, dispelling cold and relieving pain, and activating blood circulation and dredging meridians. Can be used for treating sexual impotence, cold womb, cold pain of waist and knee, asthma due to kidney deficiency, conjunctival congestion, pharyngalgia, cold pain of heart and abdomen, vomiting and diarrhea due to deficiency cold, cold hernia, amenorrhea, dysmenorrhea, etc. Pharmacological actions include: 1. has effects on cardiovascular system (tonifying heart, dilating blood vessel, lowering blood pressure); 2. acting on digestive system (water decoction inhibits small intestinal peristalsis of mice, and cassia oil promotes intestinal peristalsis, so that secretion of digestive juice is increased, and gastrointestinal cramp pain is relieved); 3. antiulcer; 4. anti-platelet aggregation, anti-coagulation; 5. exciting the pituitary-adrenal cortex system; 6. regulate body immunity.

Chinese angelica root: sweet and pungent taste, warm nature, and good effect in nourishing liver, heart and spleen meridians, and has effects of replenishing blood, promoting blood circulation, regulating menstruation, relieving pain, loosening bowel, and relieving constipation. Is mainly used for treating blood deficiency and sallow complexion, dizziness and palpitation, irregular menstruation, amenorrhea and dysmenorrhea, deficiency-cold abdominal pain, constipation due to dryness of the intestines, rheumatalgia, traumatic injury, carbuncle, ulcer and the like. Pharmacological actions include: 1. has effects on blood system (promoting hematopoiesis of bone marrow, inhibiting platelet aggregation, resisting thrombosis, reducing blood lipid, and resisting atherosclerosis); 2. has effects on cardiovascular system (anti-myocardial ischemia, anti-fibrosis after myocardial infarction, anti-arrhythmia, dilating blood vessel, lowering blood pressure); 3. exciting or inhibiting uterine smooth muscle; 4. enhancing the immune function of the organism; 5. radiation resistance; 6. damage resistance; 7. protecting liver; 8. anti-tumor.

Radix paeoniae rubra: bitter taste and slightly cold nature. Enter liver meridian. Has effects in clearing away heat and cooling blood; promoting blood circulation to remove blood stasis. Main toxic heat and spot; hematemesis and epistaxis; the wind of the intestines descends blood; conjunctival congestion with swelling and pain; carbuncle, swelling and sore; amenorrhea and dysmenorrhea; collapse of the belt and turbid urine; hypochondriac pain due to stasis; hernia mass; traumatic injury. Can be used for treating toxic heat, speckle, hematemesis, epistaxis, conjunctival congestion, swelling and pain, liver Yu Xie pain, amenorrhea, dysmenorrhea, abdominal pain, traumatic injury, carbuncle, swelling and ulcer. It is often indicated for fever bleeding, conjunctival congestion, swelling and pain, carbuncle, swelling and sore toxicity due to warm disease and heat entering blood system. Pharmacological actions include: 1. antithrombotic effect; 2. anti-platelet aggregation; 3. reducing blood lipid and resisting arteriosclerosis; 4. effects on the cardiovascular system; 5. an antitumor effect; 6. protecting liver.

Jujube: sweet in flavor and warm in nature. Enter spleen and stomach meridians. Tonify middle energizer, replenish qi, nourish blood and tranquilize mind. Can be used for treating spleen deficiency, anorexia, debilitation, loose stool, and dysphoria of women. Pharmacological actions include: 1. central inhibitory action; 2. protecting liver; 3. enhancing muscle strength and resisting fatigue; 4. anti-allergic reactions; 5. an anti-tumor; 6. tranquilizing and allaying excitement; 7. increasing leukocyte cAMP and enhancing immunity.

Akebia stem: bitter in flavor and cold in nature. It enters heart, small intestine and bladder meridians. Clear heat and induce diuresis, activate blood and promote blood circulation. Reddish urine, stranguria with turbid urine, edema, dysphoria with smothery sensation in the chest, throat pain, sore of mouth and tongue, rheumatalgia, galactostasis and amenorrhea. Pharmacological actions include: 1. urination promoting; 2. antibacterial and antifungal.

The beneficial effects of the invention are that

The invention provides a compound Chinese medicinal dreg fermentation liquor for preventing and treating fowl colibacillosis and a preparation method thereof, wherein the compound Chinese medicinal dreg fermentation liquor is prepared by fermenting lactobacillus rhamnosus PC30-2-1, and can reduce the inflammatory cytokine level of pathogenic escherichia coli of artificially infected fowl, improve the bursal index, improve the appetite and disease resistance of the fowl, promote weight gain, improve production benefits, reduce or replace the use of conventional antibacterial medicines, reduce medicine residues, reduce the selection pressure of the antibacterial medicines and inhibit or slow down the generation of clinical pathogenic bacteria resistance.

Seven traditional Chinese medicines in the traditional Chinese medicine residue prescription are all common traditional Chinese medicines, have low price, have no toxic or side effect on the bodies of the ill livestock and poultry, have no residue in the bodies of the livestock and poultry, and have no drug resistance. In vitro experiments show that the medicine has good inhibition effect on escherichia coli; in vivo experiments show that the compound Chinese medicinal residue fermentation liquor has better protection and treatment effects on artificially infected fowl pathogenic escherichia coli, and is expected to replace common antibacterial medicaments for preventing and treating livestock colibacillosis.

Drawings

FIG. 1 is a growth bar graph of mice drenched with the fermentation broth of example 2;

FIG. 2 is a bar graph of Zhou Zengchong of test chickens drenched with the fermentation broth of example 2;

FIG. 3 is a view of a sectioned organ lesion;

FIG. 4 is a bar graph of cytokines for IL-6 content;

FIG. 5 is a bar graph of IL-8 content of cytokines;

FIG. 6 is a histogram of cytokines for serum IFN-gamma content.

Detailed Description

Example 1 isolation and characterization of Lactobacillus rhamnosus PC30-2-1

(1) Separation and purification

The commercial pickle juice is placed in a sterile ultra-clean workbench, the sample is diluted 10 times in gradient by using normal saline, 100 mu L of diluent with proper gradient is evenly coated on MRS culture medium containing 2% calcium carbonate, and the coated culture medium is placed in an anaerobic tank for culturing for 48 hours at 37 ℃. Different forms of colonies with obvious calcium dissolving rings are selected and inoculated in a corresponding liquid culture medium for enrichment culture for 24 hours, then streaked and inoculated on an MRS solid culture medium again, and after separation and purification for 3 times, single colonies are selected and subjected to gram staining. Colonies positive for gram staining were subjected to enrichment culture and stored at-80℃in MRS liquid medium (containing 20% glycerol). All strains were subjected to 3 subcultures prior to use. After purification by MRS separating culture medium, round, milky white, regular edge, smooth surface, diameter of 1-1.5mm, paired or chain arranged bacterial colony, and gram staining microscopic examination to obtain dispersed and chain arranged short rod-shaped bacterial body.

(2) 16S rDNA sequencing identification

The purified strain was subjected to enrichment culture in 5mL of liquid medium for overnight, and genomic DNA of the isolated strain was extracted using a bacterial DNA genome extraction kit from Shanghai Co., ltd.

The specific fragment of the isolated strain 16S rDNA was PCR amplified by using the above genomic DNA as a template and by using the general primer (forward sequence: 27F:5'-AGAGTTTGATCMTGGCTCAG-3' and reverse sequence: 1490 2R:5'-TACGGYTACCTTG TTACGACTT-3') for bacterial 16S rDNA synthesized by Bio-company, and a 50. Mu.L PCR reaction system was shown in tables 1-2. The PCR amplification procedure was set as follows, pre-denaturation: 94 ℃ for 5min; denaturation: 94 ℃ for 30s; annealing: 55 ℃ for 30s; extension: 72 ℃ for 90s;30 cycles; terminal extension: 72 ℃ for 10min.

TABLE 1 PCR reaction System

Tab.1 Reaction system of PCR

And 5 mu L of PCR amplified products are taken, 1% agarose gel electrophoresis detection is carried out, a sample with a clear band at 1600bp is subjected to gel cutting recovery, the gel recovery products are connected with a pMD18-T cloning vector and are transformed into DH5 alpha competent cells, and positive cloning plasmids are sent to a biological engineering (Shanghai) stock company for sequencing analysis. The resulting sequencing results were compared to the data in NCBI using BLAST to determine their species. The comparison result of the 16S rDNA sequence shows that the sequence of the strain is 99 percent consistent with that of L.rhamnosus.

Example 2 preparation of Compound Chinese medicinal fermentation liquor

Step one: inoculating lactobacillus rhamnosus PC30-2-1 preserved in a laboratory into 5mL of sterilization MRS liquid culture medium according to 1% inoculation amount, and performing shake culture at 37 ℃ for 12h at 150r/min to obtain seed culture solution;

step two: weighing 12g of asarum, 12g of liquorice, 12g of cinnamon, 12g of angelica, 12g of red paeony root, 12g of jujube and 8g of akebia stem, slicing the traditional Chinese medicines, adding 5L of water, decocting for 60min to 2L, and filtering the filtrate with gauze; adding water for 3L again, boiling water, decocting with slow fire for 45min to 1L, and mixing filtrates;

step three: drying the residues, weighing a certain amount of dried residues in a conical flask, adding distilled water according to a feed-liquid ratio of 1:10, sterilizing at 121deg.C under high pressure, and cooling to room temperature;

step four: aseptically inoculating the seed culture solution obtained in the first step into the conical flask in the third step according to the volume ratio of 0.5%, performing shake culture for 18h at 37 ℃ and 150r/min, centrifuging the dreg fermentation liquor at 4 ℃ and 8000r/min for 10min, vacuum-filtering the supernatant obtained after centrifugation through a 0.45 mu m filter membrane, and concentrating the filtrate by rotary evaporation in a rotary evaporator (45 ℃ and 75 r/min) until the volume ratio is 1:3; obtaining the compound traditional Chinese medicine fermentation liquor.

EXAMPLE 3 oxford cup method test of antibacterial Activity of Compound traditional Chinese medicine fermentation liquor on E.coli Standard strain ATCC25922

Inoculating ATCC25922 strain preserved in laboratory into sterilized LB liquid medium at 1% strain, shaking culturing at 37deg.C for 150r/min to OD ₆₀₀ =0.5 (about 10 ⁸ CFU/mL), 100-fold serial dilutions were made using LB liquid medium for drug susceptibility testing;

100 mu L of the cultured ATCC25922 strain is uniformly coated on an LB solid culture medium plate, a sterilized oxford cup is uniformly placed, 200 mu L of the concentrated fermentation liquid prepared in the example 2 is respectively added into the oxford cup, and the mixture is subjected to static culture for 12 hours in a constant temperature incubator at 37 ℃;

observing whether the flat plate has a bacteriostasis ring or not, and measuring the diameter of the bacteriostasis ring by using a ruler; the oxford cup experiment result shows that the concentrated fermentation supernatant of the traditional Chinese medicine residue has a certain antibacterial effect on the escherichia coli ATCC25922 strain with the diameter of about 12 mm.

Example 4

Healthy Kunming mice (weight 20+/-2 g) are selected, and 10 mice/group are infused with the prepared dreg fermentation broth of the example 2 for 15 continuous days, 200 ul/day, and the feeding, drinking, weight change and health condition of the mice are observed;

as shown in fig. 1, fig. 1 is a growth histogram of mice drenched with the fermented liquid of the residues prepared in example 2, and as can be seen from fig. 1, the test mice were better in drinking water, and were bright by Mao Pingshun, and are psychoactive and have no abnormality after drenching with the fermented liquid of the compound Chinese medicinal residues. The weight of mice infused with 1% of the compound Chinese medicine residue fermentation liquor is obviously increased compared with the weight of mice infused with the compound Chinese medicine residue fermentation liquor in a control group 10 days before the test.

Example 5

30 healthy Rockwell chickens (with basic immunity completed) of 7 days old are selected, and the chickens are adapted to the environment for 5 days, and are set as a test group and a control group, 10 chickens/group, and male and female chickens are half. The compound traditional Chinese medicine fermentation broth of the embodiment 2 is fed for 21 days by continuously drinking water according to the addition amount of 1% and 5%, the drinking water is calculated for 2 times/day, and each group of test chickens can drink completely within 2 hours each time. On day 22 of the experiment, the addition of the fermentation broth was stopped and the bird-pathogenic E.coli CC1 strain was infected with 0.5 mL/strain (about 10 ⁹ CFU/mL). And (3) 48 hours after the bacterial attack, observing the states, clinical symptoms and the like of the test chickens, collecting venous blood under the wings, detecting the levels of cytokines such as serum IL-2, IL-6, IL-8, IFN-gamma and the like, carrying out section inspection on 6 test chickens in each group, observing organ lesions, and calculating organ indexes such as spleen, bursa of Fabricius and the like.

As shown in fig. 2, fig. 2 is a Zhou Zengchong bar graph of test chickens drenched with the prepared residue broth of example 2, and as can be seen from fig. 2, 1% and 5% of the residue broth uniformly promote weight gain to some extent in the test chickens. Wherein, the weight gain effect of the 5% addition group is optimal at the 2 nd week, the weight gain effect of the 1% addition group is optimal at the 3 rd week, but the difference with the control group is not obvious. The results show that the compound traditional Chinese medicine residue fermentation liquor is safe to test animals, has no toxic or side effect, can improve the appetite of the animals to a certain extent, promote weight gain and reduce feed conversion ratio.

Fig. 3 is a graph of organ lesion of a section, wherein a represents a control group (not administration), b represents a fermentation broth adding group, and the pericardium and liver are section-examined, and fig. 3 illustrates that after bacteria attack, the feeding water intake of each group of test chickens is reduced to different degrees in comparison with ordinary times, individual chickens in the control group only show mental depression, eye and nose secretion is increased, wings are relaxed, and the overall feeding and mental condition of the compound Chinese medicinal dreg fermentation broth adding group is slightly better than that of the control group. Individual chickens were examined only for pathological changes such as thickening and turbidity of pericardium, cellulose pericarditis, hepatomegaly, bleeding blood stasis, and cellulose exudates on the liver surface. The chickens with the symptoms are fewer in the compound Chinese medicinal residue fermentation broth added group (figure 3 b), and the overall symptoms are lighter than those in the control group (figure 3 a).

FIG. 4 is a bar graph of IL-6 content, FIG. 5 is a bar graph of IL-8 content, FIG. 6 is a bar graph of serum IFN-gamma content, FIGS. 4-6 illustrate that both 1% and 5% of the fermentation broth of the drug residue significantly reduced serum IL-6 levels compared to the control group; in addition, 1% of the Chinese medicinal residue fermentation liquor can also obviously reduce secretion of cytokines such as serum IL-8, IFN-gamma and the like, which shows that the Chinese medicinal residue fermentation liquor has a certain anti-inflammatory effect.

Table 2 shows that the compound Chinese medicinal residue fermentation liquor has an effect of promoting the development of bursa of Fabricius of test chickens by 1% and 5% of Chinese medicinal residue fermentation liquor on immune organ indexes of the test chickens as shown in Table 2, wherein the bursa indexes are higher than those of the control group, but the difference is not obvious; and 1% and 5% of the Chinese medicinal residue fermentation liquor reduce the spleen index of the test chicken, and the spleen index is also insignificant compared with the control group.

Table 2 Compound Chinese medicinal residue fermentation broth%

Claims (9)

1. A preparation method of a compound Chinese medicinal residue fermentation broth for preventing and treating colibacillosis of poultry is characterized by comprising the following steps of:

step one: lactobacillus rhamnosus PC30-2-1 is obtained by separating from commercial pickle, and strain identification is carried out through 16S rDNA, so as to obtain lactobacillus rhamnosus PC30-2-1 (Lactobacillus rhamnosus PC-2-1) with the preservation number of CCTCC NO: M2022083, wherein the preservation unit is: china center for type culture collection, with the preservation addresses: the preservation date of the university of Chinese Wuhan is: inoculating lactobacillus rhamnosus PC30-2-1 into a culture medium at 37 ℃ for 12-18h under shaking culture at 150r/min at 2022, 1 month and 17 days to obtain seed culture solution;

step two: slicing herba asari, glycyrrhrizae radix, cortex Cinnamomi, radix Angelicae sinensis, radix Paeoniae Rubra, fructus Jujubae and caulis Akebiae, decocting in water, and mixing filtrates; the traditional Chinese medicine comprises the following raw materials in proportion: 12-15 parts of asarum, 12-15 parts of liquorice, 12-15 parts of cinnamon, 12-15 parts of angelica sinensis, 12-15 parts of red paeony root, 12-15 parts of Chinese date and 8-10 parts of akebia stem;

step three: drying the dregs obtained in the second step, adding distilled water into the obtained dry dregs, sterilizing, and cooling to room temperature to obtain dregs culture solution;

step four: inoculating the seed culture solution in the first step into the residue culture solution in the third step, shake culturing at 37deg.C and 150r/min for 12-18 hr, centrifuging, and collecting fermentation supernatant to obtain compound Chinese medicinal residue fermentation liquor.

2. The method for preparing the compound Chinese medicinal residue fermentation liquor for preventing and treating fowl colibacillosis according to claim 1, wherein the culture medium in the first step is MRS liquid culture medium.

3. The method for preparing the compound Chinese medicinal dreg fermentation liquor for preventing and treating fowl colibacillosis according to claim 1, wherein the Chinese medicinal raw materials in the second step are as follows: 12 parts of asarum, 12 parts of liquorice, 12 parts of cinnamon, 12 parts of angelica sinensis, 12 parts of red paeony root, 12 parts of Chinese date and 8 parts of akebia stem.

4. The method for preparing the compound Chinese medicinal residue fermentation liquor for preventing and treating the colibacillosis of poultry according to claim 1, wherein the second decoction time is 2 times, the first decoction time is 1h, and the second decoction time is 40-45min.

5. The method for preparing the compound Chinese medicinal residue fermentation liquor for preventing and treating fowl colibacillosis according to claim 1, wherein the drying temperature in the third step is room temperature, and the drying time is 48h.

6. The method for preparing the compound Chinese medicine residue fermentation liquor for preventing and treating fowl colibacillosis according to claim 1, wherein the feed liquid ratio of the Chinese medicine residue to distilled water in the third step is 1:10.

7. The method for preparing a compound Chinese medicinal residue fermentation broth for preventing and treating fowl colibacillosis according to claim 1, wherein the sterilization temperature in the third step is 121 ℃ and the pressure is 103.4kPa.

8. The method for preparing the compound Chinese medicinal residue fermentation liquor for preventing and treating colibacillosis of poultry according to claim 1, wherein the step four of centrifugal collection comprises the following steps: centrifuging the residue fermentation broth at 4deg.C at 8000r/min for 10min, vacuum filtering the supernatant with 0.45 μm filter membrane, and concentrating the filtrate by rotary evaporation under the conditions of 45deg.C and 75r/min to volume ratio of 1:3.

9. The compound Chinese medicinal dreg fermentation broth for preventing and treating colibacillosis of birds, which is obtained by the preparation method of claim 1.