CN113101318B

CN113101318B - A Coptidis rhizoma extractive solution and its application in treating animals infected with Staphylococcus pseudointermedius

Info

Publication number: CN113101318B
Application number: CN202110596695.4A
Authority: CN
Inventors: 李宏梅; 郭慧君; 成子强; 赵于清; 朱毅松
Original assignee: Shandong Agricultural University
Current assignee: Shandong Agricultural University
Priority date: 2021-05-31
Filing date: 2021-05-31
Publication date: 2022-08-30
Anticipated expiration: 2041-05-31
Also published as: CN113101318A

Abstract

The invention discloses a coptis chinensis extracting solution and application thereof in treating animals infected by staphylococcus pseudointermedium. The research of the invention finds that the coptis chinensis extracting solution has obvious bacteriostatic activity on the staphylococcus pseudointermedium and does not generate drug resistance. The coptis chinensis extract prepared by the method can effectively treat diseases such as skin soft tissue injury and the like caused by infection of pseudo-intermediate staphylococcus to animals by continuously taking the extract for 5d to 7d according to 2-5 ml/kg BW, the cure rate reaches 100 percent, and the extract has no drug residue, toxic or side effect and no drug resistance.

Description

A Coptidis rhizoma extractive solution and its application in treating animals infected with Staphylococcus pseudointermedius

Technical Field

The invention relates to the technical field of pseudo-intermediate staphylococcus infection treatment, in particular to a coptis chinensis extracting solution and application thereof in treating animals infected by pseudo-intermediate staphylococcus.

Background

The Staphylococcus pseudointermedius is a novel Staphylococcus discovered and named in 2005 by Devriese and the like, is a gram-positive facultative anaerobe, mainly inhabits dogs and cats companion animals, is also reported to be infected by foxes, rabbits, fur-bearing animals, humans and the like, is part of a normal skin microflora, and is a common symbiotic bacterial pathogen on skin and mucosa.

Staphylococcus pseudomedians is a opportunistic pathogen that causes invasive infections and is found in a wide range of animal skin, hair follicles, coat, nasopharyngeal mucosa, and intestinal tracts, especially in the skin mucosa areas such as the mouth, nose, groin, perineum, and anus. When the resistance of the host is reduced and the skin barrier is changed by susceptible factors, so that a series of opportunistic infections of the animal body can be caused, and the chest, the abdomen and the limbs are caused, wherein the symptoms of subcutaneous tissue swelling, suppuration, ulceration and skin scabbing are shown, and the serious patients flow off yellow and white secretion or emit rancid odor. In addition to skin infections, there are other infections such as otitis externa, urinary tract infections, postoperative infections, bacteremia, etc.

The most common clinical manifestations of canine infection with staphylococcus pseudointermedia are superficial bacterial folliculitis and atopic dermatitis; on postnatal day 1, pathogens can be isolated from the skin and mucous membranes. The clinical symptoms of human infection are local skin lesions such as folliculitis, cellulitis and the like, and severe patients even have bacteremia, brain abscess, endocarditis or pneumonia, bacteremia and the like. Raccoon dogs are mainly characterized by skin suppuration and respiratory dysfunction, and the suppuration mainly occurs in joints, neck and inguinal area and is pustules filled with yellowish white pus, blood and tissue fluid. Respiratory dysfunction is manifested by cough, sneezing, asthma, shortness of breath, cyanosis of mucous membrane in abdominal respiration, mucus secretion in nasal cavity, and purulent nasal discharge. Also can cause various animal diseases such as cow mastitis and endometritis, and is considered as a main animal-derived pathogenic bacterium.

The staphylococcus pseudointermedia belongs to the staphylococcus intermedia (including staphylococcus intermedia, staphylococcus dolphin and staphylococcus pseudointermedia), is similar to the staphylococcus intermedia, but is different from the staphylococcus intermedia and the staphylococcus dolphin in biochemical characteristics, evolutionary relations and the like (the staphylococcus pseudointermedia can make maltose and trehalose positive and mannitol negative, the staphylococcus intermedia can make trehalose positive and maltose and mannitol negative, and the staphylococcus dolphin can make maltose and mannitol positive and trehalose negative). The staphylococcus intermedium and the staphylococcus aureus are greatly different in culture characteristics, biochemical characteristics, pathogenic factors, evolutionary relationships and the like, and are non-congeneric staphylococcus.

Staphylococcus pseudointermedius can be classified into methicillin-sensitive Staphylococcus pseudointermedius (MSSP) and methicillin-resistant Staphylococcus pseudointermedius (MRSP) according to the drug resistance characteristics. Besides the resistance to beta-lactam antibiotics, MRSP also exhibits resistance to most of the antibacterial agents on the market, such as macrolides, lincosamides, aminoglycosides, tetracyclines, chloramphenides, quinolones, and the like. The incidence of MRSP has increased since 2006, becoming an important animal health problem in the veterinary field.

Repeated use of antibiotics by animals with recurrent pyoderma has a strong bearing on the rapid global spread of MRSP. The growing spread of multidrug resistance (MDR) and MRSP strains has greatly limited the therapeutic options for bacterial skin infections, making the treatment of MRSP infections a major challenge in the veterinary field. Overall, the potential impact of MRSP on animal health, human health, animal production is enormous.

MRSP colonizes healthy animals and animal workers who may become carriers of MRSP, thereby facilitating the spread of MRSP among animal and human populations. It has been reported that dogs with MRSP can carry over one year after clinically significant infection, and after antibiotic treatment dogs have been sampled after the treatment, MRSP is still found to be positive, so that there is an urgent need for alternative antibiotic therapy to treat drug-resistant Staphylococcus pseudointermedius infection.

Disclosure of Invention

In view of the prior art, the invention aims to provide a coptis chinensis extract and application thereof in treating animals such as dogs and cats infected by staphylococcus pseudointermedium. The research of the invention finds that the coptis chinensis extracting solution has obvious bacteriostatic activity on the staphylococcus pseudointermedium, does not generate drug resistance, and can effectively treat diseases caused by the infection of the staphylococcus pseudointermedium on animals.

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

in a first aspect of the invention, the application of the coptis chinensis extract in the following (1) or (2) is provided:

(1) preparing an antibacterial product for inhibiting the proliferation of the pseudo-intermediate staphylococcus;

(2) preparing a medicament for treating diseases caused by infection of the staphylococcus pseudointermedium.

Preferably, the staphylococcus pseudointermedia is methicillin-resistant staphylococcus pseudointermedia (MRSP).

Preferably, the disease caused by infection with Staphylococcus pseudointermedius comprises: the animal skin soft tissue injury, suppuration, ulcer, pneumonia, endocarditis, brain abscess, urinary tract infection, bacteremia and the like.

Preferably, the medicament can be in the form of oral liquid, injection, tablets, capsules, granules and the like.

Preferably, the coptis chinensis extract is prepared by the following method:

soaking Coptidis rhizoma in 10-15 times of water for 30-60min, decocting for 20-30min, filtering, and separating residue to obtain first filtrate;

adding 5-10 times of water into the residue, decocting for 15-20min, and filtering to obtain a second filtrate;

combining the first filtrate and the second filtrate, and concentrating until the weight of the filtrate is equal to the weight of the initially added coptis chinensis, so as to obtain a concentrated filtrate;

centrifuging the concentrated filtrate, separating supernatant, and aseptically filtering or sterilizing the supernatant to obtain Coptidis rhizoma extractive solution.

Preferably, the concentration of the coptis extract is more than or equal to 1 g/ml.

The second aspect of the invention provides a traditional Chinese medicine oral liquid for treating pseudo-intermediate staphylococcus infection, which takes coptis chinensis extract as an active ingredient.

Preferably, the coptis chinensis extract is prepared by the following method:

In a third aspect of the present invention, there is provided a method for treating a Staphylococcus pseudointermedia infection, which comprises administering to a subject at risk of or diagnosed with a disease caused by the Staphylococcus pseudointermedia infection a therapeutically effective amount of the above Coptidis rhizoma extract.

The actual dosage to be used may be determined according to the relevant circumstances. These include: the physiological state of the subject, the route of administration, age, weight, individual response to the drug, severity of the symptoms, and the like. Illustratively, the therapeutically effective amount may be 2-5 ml/kg. BW/time, 1 time/day, for 5-7 consecutive days.

The invention has the beneficial effects that:

the coptis chinensis extract prepared by the method has the effects of obviously inhibiting the growth of the staphylococcus pseudointermedium, resisting the infection of the staphylococcus pseudointermedium and regulating the immune function, can effectively treat diseases caused by the infection of the staphylococcus pseudointermedium, and has no drug residue, no toxic or side effect and no drug resistance. The test proves that: the coptis chinensis extract prepared by the method can effectively treat diseases such as skin soft tissue injury and the like caused by pseudo-intermediate staphylococcus infected animals after being continuously orally taken for 5d to 7d according to 2 to 5ml/kg of BW, and the cure rate reaches 100 percent.

Drawings

FIG. 1: the coptis chinensis extracting solution has an antibacterial effect on pseudo-intermediate staphylococcus; in the figure, No. 1, coptis extract; number 2, scutellaria baicalensis extracting solution; number 3, forsythia suspensa extract; no. 4, saline.

FIG. 2: the bacteriostatic effects of different products on the pseudo-intermediate staphylococcus are respectively 18h (left) and 24h (right); in the figure, No. 1, meropenem; no. 2, coptis extract; number 3, coptidis rhizoma shangqing pills; no. 4, SHUANGHUANGLIAN oral liquid; and 5, normal saline.

FIG. 3: the Minimum Inhibitory Concentration (MIC) of Coptidis rhizoma extract.

FIG. 4 is a schematic view of: and (4) measuring the MIC of the rhizoma coptidis liquid by a seed dibbling method.

FIG. 5: comparison of skin soft tissue lesions before and after treatment of mice infected with Staphylococcus pseudointermedius (A, before treatment; B, after treatment).

FIG. 6: the skin soft tissue lesions of the red raccoon dog infected with the staphylococcus pseudointermedius before and after administration are compared (A, before administration; B, after administration).

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

As mentioned in the background section, staphylococcus pseudointermedia is a new type of staphylococcus discovered only in recent years, and mainly infects canine and feline companion animals, fur-bearing animals such as foxes and raccoon dogs, and the like, and can cause serious diseases and losses. Compared with the existing common pathogenic bacteria staphylococcus aureus, the bacillus subtilis has obvious differences in culture characteristics, biochemical characteristics, pathogenicity and the like. At present, few reports about staphylococcus pseudointermedia exist, and the technical maturity of related researches is low.

Pseudointermediate staphylococci, particularly oxacillin-resistant pseudointermediate staphylococci (MRSP), exhibit resistance to most of the antibacterial agents on the market, such as macrolides, lincosamides, aminoglycosides, tetracyclines, chloramphenics, quinolones, and the like. Therefore, it is extremely difficult to control infection caused by Staphylococcus pseudointermedius.

In the research process, the invention unexpectedly discovers that the coptis extract as the only active component can obviously inhibit the growth of the staphylococcus pseudointermedium, and more importantly, the coptis extract can not generate drug resistance. The coptis chinensis extracting solution does not need to be matched with other Chinese herbal medicines or components, has the advantages of definite effect, more definite active ingredients and lower cost compared with a compound prescription, and is more convenient and more favorable for preparing an injection in the future.

In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.

The test materials used in the examples of the present invention were all conventional in the art and commercially available.

Example 1: preparation of Coptidis rhizoma extract

(1) Pulverizing Coptidis rhizoma 100g, soaking in 1.5L water for 45min, decocting, boiling with strong fire, decocting with slow fire for 20min, and filtering with 2 layers of sterile gauze to obtain filtrate.

(2) Adding water into the left residue to 1000mL, decocting the residue again to boil, filtering the liquid medicine with gauze, and discarding the residue.

(3) And (3) combining the filtrates in the step (1) and the step (2), concentrating to 100mL by using a small fire to obtain a concentrated filtrate, and cooling to room temperature.

(4) Centrifuging the concentrated filtrate with a centrifuge at 1200 rpm for 6 minutes, collecting supernatant, filtering with a filter on a super clean bench under sterile condition to obtain Coptidis rhizoma extractive solution with concentration of 1g/mL, and storing in a refrigerator.

Example 2: in vitro bacteriostasis test

1. Bacteriostatic investigation of coptis extract on pseudo-intermediate staphylococcus

A prepared staphylococcus pseudointermedium (staphylococcus pseudointermedium strain is separated and identified from a laboratory published paper Zhu et al,2020, DOI:10.1111/tbed.13847, and a test with a duplicate can be obtained by Shandong agriculture university) suspension is evenly inoculated on a TSA agar plate culture medium by using an inoculation glass rod, a medicine name mark is made on the plate culture medium, a sterilized Oxford cup is placed on a corresponding medicine mark position under an ultraclean workbench, a slight press is carried out to ensure that no gap is left on the contact surface of the Oxford cup and the solid culture medium, then 200 mu L of coptis extract, scutellaria extract and forsythia extract with the mass concentration of 1g/mL are respectively sucked by a liquid transfer gun, and the preparation method of the scutellaria extract and the forsythia extract is the same as that in example 1, except that the raw material coptis chinensis is replaced by scutellaria baicalensis or forsythia suspensa and the physiological saline solution of a negative control group is injected into the Oxford cup, and (3) gently transferring to a 37 ℃ incubator for culturing for 18-24 h, measuring the size of the inhibition zone by using a vernier caliper for 18h and 24h respectively, and repeatedly measuring the diameter of the inhibition zone of each medicament for multiple times to obtain an average value.

The determination result of 24h is shown in figure 1, and the result shows that the bacteriostatic circle of the coptis extract (No. 1) can reach 25 +/-0.5 mm, and the bacteriostatic effect is close to that of the antibiotic meropenem. The antibacterial zones of the other two Chinese herbal medicine radix Scutellariae extract (No. 2), fructus forsythiae extract (No. 3) and normal saline (No. 4) are respectively 10 + -0.1 mm, 16 + -0.5 mm and 0 + -0.0 mm.

2. Different products have a comparison on the in vitro bacteriostatic effect of the pseudo-intermediate staphylococcus:

the primary screening of the coptis extract with better antibacterial effect is compared with the in vitro antibacterial effect of the commercial Shuanghuanglian oral liquid, the coptis supernatant pill and the meropenem respectively, and the antibacterial result after culturing for 18h is shown in figure 2. The Meropenem, the coptis extract and the coptis supernatant pill are found to have the antibacterial effect from strong to weak, the average antibacterial circle diameters are 26 +/-0.8 mm, 25 +/-0.5 mm and 11 +/-0.3 mm respectively, and the Shuanghuanglian oral liquid has almost no antibacterial effect. And continuing culturing for 24h, and finding that a new bacterial colony begins to appear in the meropenem bacteriostatic zone, and the situation of growing bacteria indicates that the drug resistance is generated, but no new bacterial colony appears in the coptis chinensis extract bacteriostatic zone, which indicates that the pseudo-intermediate staphylococcus has the drug resistance to the meropenem and has no drug resistance to the coptis chinensis extract.

3. And (3) determining the Minimum Inhibitory Concentration (MIC) of the coptis extract to the staphylococcus pseudointermedium:

taking 11 sterile test tubes with rubber plugs, and marking the test tubes with the numbers of 1-11; adding 3mL of TSB liquid culture medium into each tube, adding 3mL (with the concentration of 1g/mL) of coptis extract into the 1 st test tube, uniformly mixing, sucking 3mL into the 2 nd tube, repeating the above operations until reaching the 9 th tube, uniformly mixing, and discarding 3 mL; no drug solution was added as a positive control in the 10 th tube; the diluted staphylococcus pseudointermedia bacterial liquid (the concentration is 1 multiplied by 10) is sucked ⁷ cfu/mL) of the culture medium is added into No. 1 to No. 10 tubes respectively, No. 11 tubes are not inoculated with bacteria, and the same amount of normal saline is added as a negative control, and then the culture medium is placed in a water-proof constant temperature incubator at 37 ℃ for culture for 12 to 16 hours. Observing the test result, because the color of the Chinese medicinal liquid is dark and the minimum inhibitory concentration can not be judged by observing the clarity of each test tube liquid with naked eyes, adopting a dibbling method to dibble 7 mu L of each test tube liquid on a solid culture medium, and culturing at 35 DEG CAfter 24h, the presence or absence of bacterial growth was observed, and the presence or absence of bacterial growth was marked as "+" and the absence of bacterial growth was marked as "-". The MIC value of the test drug was determined as the minimum drug concentration for no bacterial growth.

FIG. 3 shows that the dilution ratio of the 1 st to 9 th wells is 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:128, 1:256, 1:512g/mL, i.e. the drug concentration is 500mg/mL, 250mg/mL, 125mg/mL, 62.5mg/mL, 31.25mg/mL, 15.63mg/mL, 7.81mg/mL, 3.91mg/mL, 2.0mg/mL respectively; FIG. 4 shows the results of the measurement of the Minimum Inhibitory Concentration (MIC) of Coptidis rhizoma by the dibbling method to be 7.81mg/mL (1:128 dilution ratio).

Example 3: infection treatment test in mice

Healthy Kunming mice with 22 g-30 g weight at 4 weeks of age were randomly divided into 4 groups: the traditional Chinese medicine comprises a traditional Chinese medicine group, an antibiotic group, a positive control group and a negative control group, wherein each group comprises 6. Firstly, infection test is carried out, wherein a negative control group is inoculated with 0.2mL of sterilized TSB culture medium in each abdominal cavity, and a positive control group, a traditional Chinese medicine group and an antibiotic group are inoculated with 1 multiplied by 10 in each abdominal cavity of each mouse ⁹ CFU/mL 0.2mL pseudo-middle staphylococcal liquid, once daily, continuously inoculated for 7 days, continuously observed for 2 w. Treatment trials were performed on skin soft tissue lesions: the traditional Chinese medicine is administrated by drenching 0.2mL of 1g/mL coptis extract for each mouse, 0.2mL of 0.1g/mL meropenem antibiotic solution is administrated by drenching 0.2mL of sterilized TSB culture medium for each mouse of the antibiotic group and the positive control group and the negative control group, the traditional Chinese medicine is administrated once a day for 7 days continuously, 2w is observed, and samples are taken to detect various evaluation indexes.

Continuously observing for 14d after inoculation and infection, wherein no symptoms or damage occurs to the negative control group mice; the positive control group, the traditional Chinese medicine group and the antibiotic group have symptoms such as slow movement, sleepiness, depression, inappetence, eye closure, necking, heavy eyes, fear of coldness, sleepiness, squatting in a cage, wrinkle of fur and the like, most of the mice have symptoms such as belly skin ulceration and suppuration with different degrees of severity, one of the mice has belly skin ulceration, and part of intestinal tract with serious adhesion can be seen in the eye; some mice had smaller suppuration foci and were less suppurative (fig. 5A). Separating and culturing ulcerative skin pus of infected mice, detecting gram-positive cocci by smear microscopy, and identifying by 16SrDNA, wherein the separated strains are consistent with the inoculated strains and are all pseudo-intermediate staphylococci. After treatment, mice in the antibiotic group and the traditional Chinese medicine group had increased appetite, increased activity, increased body weight, better mental status, gradual healing of skin ulcer and suppuration, and the like, and did not die (fig. 5B).

Statistics are carried out on the infection rate, the death rate and the cure rate of mice infected with staphylococcus pseudointermedia before and after treatment, and the results are shown in table 1.

TABLE 1 infection, mortality, and cure rates of mice infected with Staphylococcus pseudointermedius before and after drug treatment

The result shows that the infection rate of the traditional Chinese medicine and the antibiotic group is 100 percent and the death rate is 0 percent; after treatment, the infection rate is 0%, the death rate is 0%, and thus the cure rates of the two groups are 100%. While the infection rate of the positive control group is 100%, all mice do not have improved disease condition and even almost half of mice die. One reason is that the body of the mice has a reduced resistance to bacterial invasion, and on the other hand, it is stated that if left untreated, a Staphylococcus pseudomedians infection can lead to severe ulceration and suppuration of the soft tissues of the skin, and even death.

The body weight, wound diameter and lesion number of mice infected with staphylococcus pseudointermedia before and after treatment were counted, and the results are shown in table 2.

TABLE 2 comparison of body weight, wound diameter, lesion number of mice infected with Staphylococcus pseudointermedius before and after drug treatment

Note: the different letters in the shoulder note indicate significant difference (P <0.05), the same letter designation indicates insignificant difference (P > 0.05).

The results show that: at 14d after infection of staphylococcus pseudointermedius, the weight of mice in the positive control group, the traditional Chinese medicine group and the antibiotic group is remarkably reduced (P is less than 0.05) relative to that in the negative control group, and the three groups have no remarkable difference; after the administration treatment, the weight of the mice of the traditional Chinese medicine group and the antibiotic group is remarkably increased compared with that of the positive control group (P <0.05), but is still lower than that of the negative control group. Mice infected with staphylococcus intermedia 14d developed ulcer lesions of varying sizes on the abdominal and leg skins. The detection shows that the number of skin ulcer lesions of each mouse of the positive control group, the antibiotic group and the traditional Chinese medicine is between 1 and 3, and the ulcer area is from the size of soybean grains to the size of adult nails and is continuously enlarged. The mice treated by the drug have the lesions disappeared and the skin healed, while the number and the size of the lesions of the untreated positive control mice are increased.

Anticoagulated blood is collected before and after treatment of mice infected with the staphylococcus pseudointermedius, and blood routine detection is carried out. The results are shown in Table 3.

TABLE 3 results of routine blood tests of mice infected with Staphylococcus pseudointermedius before and after drug treatment

As a result, the mice infected with 14d had a significant increase in leukocytes and the like (P <0.05), and the erythrocytes, hemoglobin and platelets all had a significant decrease (P <0.05), showing significant symptoms such as bacterial infection and anemia. After 14 days of treatment by using the coptis chinensis extract and the antibiotics, the indexes are restored to a certain degree, and the symptoms are greatly relieved, wherein the white blood cells, the lymphocytes and the neutrophils in the traditional Chinese medicine are more obviously reduced than those in the antibiotic group, and the restoration speed and the effect are obviously better than those in the antibiotic group; the recovery of the number of red blood cells and the content of hemoglobin of the traditional Chinese medicine is faster than that of the antibiotic group.

Example 4: canidae animal red raccoon dog infection treatment test

12 red raccoon dogs aged for 95 days are taken alongThe machine is divided into 3 groups: 4 positive control groups, 4 traditional Chinese medicine groups and 4 negative control groups. The positive control group and the traditional Chinese medicine group are inoculated to each raccoon dog by 1 x 10 subcutaneous inoculation at the neck part of 2mL/kg BW ⁹ And (3) inoculating the CFU/mL pseudo-intermediate staphylococcus liquid and a sterilized TSB culture medium with the same dose to the neck part of the negative control group subcutaneously, inoculating once a day and continuously inoculating for 7 days, and recording skin soft tissue lesions of the abdomen, the legs and the like. After 2w, the traditional Chinese medicine group is irrigated with 1g/mL coptis extract according to 2mL/kg & BW, and each raccoon dog is irrigated once a day for 7 days continuously. The positive control group and the negative control group are drenched with the death sterilization TSB culture medium with the same dose.

After the artificial infection of the red raccoon dog 14d, soft tissue ulcer and suppuration lesions similar to those of the infected mouse appear on the skin of the abdomen and the legs, and the result is shown in fig. 6A. After the coptis chinensis extract is infused, the soft tissue ulcer and suppuration of 14d raccoon skin are healed, the tissue of the pathological change part is proliferated, and new hair appears; the untreated raccoon skin soft tissue ulceration and suppuration were further enlarged or worsened, and the results are shown in fig. 6B.

The infection rate, the death rate and the cure rate of the red raccoon dog infected with the staphylococcus pseudomedians before and after treatment are counted, and the results are shown in table 4.

TABLE 4 infection rate, mortality rate, and cure rate of racoon dogs infected with Staphylococcus pseudointermedius before and after the treatment of the Coptidis rhizoma extractive solution

Statistics shows that the infection rate of the positive control group of the raccoon dog in a treatment experiment is 100 percent, and the death rate is 0 percent; the infection rate of the traditional Chinese medicine group before treatment is 100%, the death rate is 0%, and the cure rate after treatment by the traditional Chinese medicine is 100%; the negative control group was consistently free of infection and death.

The weight, the wound diameter and the number of pathological changes of the red raccoon dog infected with staphylococcus pseudomedians before and after treatment are counted, and the results are shown in table 5.

TABLE 5 comparison of racoon dog body weight, wound diameter, and lesion number infected with Staphylococcus pseudointermedius before and after treatment with Coptidis rhizoma extract

The results show that 14d after infection of the staphylococcus intermedius, the raccoon dog has serious weight reduction and the increase of skin ulcer or suppurative wound (P < 0.05); after the coptis chinensis extracting solution is used for treatment, the weight of 14d raccoon dogs is obviously increased (P is less than 0.05), skin ulcers or suppurative wounds are reduced, and all wound lesions disappear; the weight of the raccoon dog of the untreated group is continuously reduced, the diameter of the wound is continuously enlarged, the number of the lesion wound is also obviously increased, and the infection is further worsened.

Collecting anticoagulated blood before and after treatment of the red raccoon dog infected with the pseudo-intermediate staphylococcus, and performing conventional blood detection. The results are shown in Table 6.

TABLE 6 racoon dog blood routine test result of infection with staphylococcus pseudointermedia before and after treatment of Chinese herbal medicine extract of coptis chinensis

From the results in table 6, it can be seen that 14d after infection with staphylococcus pseudointermedia, the numbers of leukocytes, lymphocytes and neutrophils in the blood of the raccoon dog are obviously increased (P < 0.05); significant decreases in red blood cell number, hemoglobin concentration, platelet number (P <0.05) appeared, indicating that severe bacterial infection induced significant white blood cell anti-infective responses and anemia symptoms. After the coptis chinensis extract is used for treatment, the number of white blood cells, the number of lymphocytes and the number of neutrophils is obviously reduced, and the anti-infection reaction of the white blood cells is obviously reduced (P is less than 0.05); the red blood cells, the thrombocytes and the hemoglobin are obviously improved (P is less than 0.05), and the anemia symptom is relieved or disappeared, so that the coptis chinensis extracting solution has an obvious relieving or treating effect on the staphylococcus pseudointermedium infection.

The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. The application of the coptis extract as the only active component in the following (1) or (2):

(2) preparing a medicament for treating diseases caused by infection of staphylococcus pseudointermedius;

the staphylococcus pseudointermedia is methicillin-resistant staphylococcus pseudointermedia;

the diseases caused by the infection of the staphylococcus pseudointermedium are animal skin soft tissue injury, suppuration, ulcer, bacteremia, leucocyte anti-infection reaction or anemia;

the coptis chinensis extracting solution is prepared by the following method: soaking Coptidis rhizoma in 10-15 times of water for 30-60min, decocting for 20-30min, filtering, and separating residue to obtain first filtrate; adding 5-10 times of water into the residue, decocting for 15-20min, and filtering to obtain a second filtrate; combining the first filtrate and the second filtrate, and concentrating until the weight of the filtrate is equal to the weight of the initially added coptis chinensis, so as to obtain a concentrated filtrate; centrifuging the concentrated filtrate, separating supernatant, and aseptically filtering or sterilizing the supernatant to obtain Coptidis rhizoma extractive solution; the concentration of the coptis extract is more than or equal to 1 g/L.

2. The use of claim 1, wherein the medicament is in the form of oral liquid, injection, tablet, capsule or granule.