CN103989725A - Acalypha australis L. and amikacin containing compound medicine for livestock and poultry - Google Patents

Abstract

The invention relates to a compound medicine for livestock and poultry containing sarcophagus and amikacin, which is composed of the following raw materials in parts by weight: 3-30 parts of amikacin and 5-200 parts of sarcophagus. Studies have shown that the addition of succulents to antibacterial drugs can reverse the resistance of bacteria to antibacterial drugs, thereby improving the therapeutic effect of antibacterial drugs. The compound drug is made into powder, tablet, oral liquid or granule according to conventional methods, and added to livestock and poultry feed according to the addition amount of 0.1-7g main drug/kg feed, which can greatly reduce the cost of drug use and drug residues , to create higher economic benefits for farmers, but also to ensure food safety.

Description

A kind of compound medicine for livestock and poultry containing Zhazhucai and amikacin

technical field

The invention belongs to the technical field of treating Escherichia coli and Salmonella infection in livestock and poultry, and in particular relates to a compound medicine for livestock and poultry containing Guangxi Yao medicine Herba chinensis and amikacin.

Background technique

Avian salmonellosis is a general term for poultry diseases caused by one or more Salmonella in the genus Salmonella. According to the different antigenic structures of pathogens, they can be divided into three categories: pullorum, avian typhoid and avian paratyphoid. Among them, Pullorum pullorum and Salmonella typhi have host specificity, mainly causing disease in chickens and turkeys; Salmonella avian paratyphi can widely infect a variety of animals and humans, and poultry and their products contaminated by it are human Salmonella infection and food poisoning one of the main sources of . Avian salmonellosis often forms a rather complex cycle of transmission. Sick poultry and infected poultry are the main sources of infection, which can be transmitted horizontally through the digestive tract, respiratory tract or conjunctiva, and can infect young poultry after vertical transmission through eggs, causing most of them to die. Use this as a breeding egg to hatch, then it can be passed on from generation to generation again and again, causing poultry salmonellosis to be difficult to purify in flocks, causing serious economic losses. Because Salmonella has a large pollution area in chicken flocks and is prone to drug resistance, the vaccines currently on the market have unsatisfactory immune effects. Therefore, quarantine of breeder flocks, drug control, environmental disinfection and purification, strengthening feeding management and sanitation and epidemic prevention are still fundamental measures to control and prevent Salmonella from entering the food chain. After hatching, the chicks drink 0.01% potassium permanganate solution or other drugs for 1 to 2 days for drug prevention. The bacteria are resistant to commonly used drugs, forcing the production of various drugs to be used alternately in order to better control drug-resistant bacteria. At present, the bacteria have strong drug resistance to aminoglycoside drugs. Aminoglycoside inactivating enzymes modify some groups necessary to protect antibacterial activity in these antibacterial drug molecules, so that they can interact with the target ribosome affinity is greatly reduced. These inactivating enzymes include aminoglycoside acyltransferase (AAC), aminoglycoside adenylyltransferase (AAD), aminoglycoside phosphotransferase (APH), adenosylnucleosyltransferase (ANT), and the like. At present, inactivating enzymes with different characteristics have been found in many aminoglycoside-resistant bacteria.

Escherichia coli is a common conditional pathogen and a common disease. It can cause acute sepsis, peritonitis, hepatitis, pneumonia, enteritis and other severe infections in chickens, and can also be secondary or mixed with viral diseases. , causing huge economic losses to the farming industry. Amoxicillin, ampicillin, ceftiofur, ceftriaxone, amoxicillin/clavulanic acid, ampicillin/sulbactam and other β-lactam ring antibiotics are commonly used important drugs for the prevention and treatment of this disease. The degree of scale intensification is increasing, and the use of drugs is becoming more and more widespread. There are more and more drug-resistant strains, and there are more reports on the drug resistance of E. coli. Studies have confirmed that one of the main mechanisms of drug resistance is the production of β-lactamase, especially extended-spectrum β-lactamase (Extended-Spectrum β-lactamase, ESBLs), which can strongly and rapidly hydrolyze β-lactamase. The amide ring makes these antibiotics lose their antibacterial activity. ESBLs are the most important enzyme type of β-lactamases. It is not only resistant to third-generation cephalosporins and aztreonam, but also cross-resistant to aminoglycosides, quinolones, and sulfonamides. It is only resistant to carbapenems, cephalosporins, and Sensitivity to mycophenes. Many literatures have reported that the multidrug resistance of Escherichia coli is closely related to the production of ESBLs, which has attracted more attention at home and abroad. The inventor has recently carried out studies on the detection and extraction of food animal-derived pathogenic bacteria ESBLs, the hydrolysis rate of enzymes on antibiotics, and the enzyme-inhibiting protective effects of sulbactam, tazobactam and traditional Chinese medicines. The drug sensitivity of some ESBLs Escherichia coli Tests showed that its multi-drug resistance rate was significantly higher than that of non-ESBLs strains. In addition, the results also showed that ESBLs-producing strains were not only severely resistant to third-generation cephalosporins such as ceftiofur, but also resistant to fluoroquinolones, aminoglycosides, sulfonamides, and fosfomycin. Severe multidrug resistance. The inventor isolated fosfomycin-resistant Escherichia coli producing the fosA3 drug-resistant gene from ESBLs-producing strains, which further enhanced the drug resistance of the bacteria.

The drug resistance of Salmonella and Escherichia coli increases the cost of medication for farmers, prolongs the course of treatment, increases the mortality of livestock and poultry, brings greater economic losses to production, and increases the risk of drug residues in livestock and poultry. Therefore, in actual production, there is an urgent need for a drug that can have a better therapeutic effect on Salmonella and Escherichia coli no matter whether it is drug-resistant or not, and can shorten the course of treatment, reduce mortality, reduce economic losses and reduce drug residues. Create higher economic benefits and protect human health and safety.

Contents of the invention

The technical problem to be solved by the present invention is to provide a compound drug for livestock and poultry containing Guangxi Yao medicine Herba chinensis and amikacin, which can kill bacteria faster and overcome the problem of antibacterial treatment caused by drug resistance. The defects of poor curative effect, large dosage, high residue and high cost avoid the problems of narrow bactericidal spectrum and slow bactericidal effect of simple traditional Chinese medicine.

In order to solve the above technical problems, the present invention adopts the following technical scheme: a compound drug for livestock and poultry containing Herba Amaranthus and Amikacin, which consists of the following raw materials in parts by weight: 3-30 parts of Amikacin, Grass 5-200 parts.

The compound medicine is prepared into powder, tablet, oral liquid or granule.

The preparation method of the compound medicine is as follows: pulverize and pass through a 60-mesh sieve, then mix with powdered antibacterial medicine, and prepare powder, tablet, oral liquid or granule according to the conventional method.

Acalypha australis L., also known as Acalypha australis L., also known as Acalypha australis L., belongs to the genus Acalyphaceae of Euphorbiaceae, herb to subshrub, with slender stems, less than 3 mm in diameter, and bract leaves of female flowers Shaped, large and leaf-shaped, up to 1 cm long, undivided, serrated at the edge, heart-shaped at the base, kidney-shaped when the female flower is open, and clam-shell-shaped when closed, with fruits hidden in it, so there is "sea clam containing The reputation of "bead" is also easy to identify. Produced in various parts of Guangxi, born on wasteland or in humid places near villages. Oral administration of whole herb decoction can treat bacillary dysentery, diarrhea and so on.

Aiming at the problem of drug resistance existing in Escherichia coli and Salmonella infection in livestock and poultry at present, the inventors use Herba chinensis and antibacterial drugs to form a compound drug. Studies have shown that the addition of succulents to antibacterial drugs can reverse the resistance of bacteria to antibacterial drugs, thereby improving the therapeutic effect of antibacterial drugs. The compound drug is made into powder, tablet, oral liquid or granule according to conventional methods, and added to livestock and poultry feed according to the addition amount of 0.1-7g main drug/kg feed, which can greatly reduce the cost of drug use and drug residues , to create higher economic benefits for farmers, but also to ensure food safety.

Detailed ways

1. Study on the reversal of antimicrobial resistance of C.

Antibacterial drugs themselves have the effect of treating Escherichia coli and Salmonella, but due to the strong drug resistance of Escherichia coli and Salmonella producing drug resistance genes, antibacterial drugs do not show a good therapeutic effect under normal dosage conditions .

1. Antimicrobial resistance inhibition test of Escherichia coli after mixed with antibacterial drugs

The minimum inhibitory concentration of antibacterial drugs against Escherichia coli strains producing ESBLs and fosA3 drug-resistant genes and the effect of E. Sub-inhibitory concentration, and then add the sub-inhibitory concentration in the culture medium, and then use the micro test tube double dilution method to culture and determine the minimum inhibitory concentration of antibacterial drugs; use the continuous passage method to add the same sub-inhibitory concentration Subculture Escherichia coli strains producing ESBLs and fosA3 drug-resistant genes in the culture medium containing Herba chinensis and the same concentration of antibacterial drugs until the observed minimum inhibitory concentration of antibacterial drugs is more than 4 times smaller than that before subculture. It is judged that Herba chinensis has drug resistance reversing effect on antibacterial drugs. The statistics of the test results are shown in Table 1:

Table 1 The subinhibitory concentration (0.5g/mL) of subinhibitory concentration (0.5g/mL) of E. coli producing ESBLs and fosA3 Escherichia coli western medicine MIC changes (μg/mL).

It can be seen from the above table that the minimum inhibitory concentration of antibacterial drugs against drug-resistant Escherichia coli was reduced by more than 4 times, and it can be judged that the grass has a drug resistance reversal effect on ESBLs and fosA3 drug-resistant Escherichia coli.

2. Antimicrobial resistance inhibition test of Salmonella after mixing Herba chinensis and antibacterial drugs

On the 96-well plate, the minimum inhibitory concentration of antibacterial drugs against drug-resistant Salmonella strains and the sub-inhibitory concentration of A. The antimicrobial concentration of the grass was cultured again using the micro test tube double dilution method to determine the minimum inhibitory concentration of the antibacterial drug; using the continuous passage method in the culture medium with the same sub-inhibitory concentration and the same concentration of the antibacterial drug The Salmonella drug-resistant strains were subcultured until the observed minimum inhibitory concentration of antibacterial drugs was more than 4 times smaller than that before the subculture, and it could be judged that the grass had a drug-resistant reversal effect on antibacterial drugs. The statistics of the test results are shown in Table 2:

Table 2 Subinhibitory concentration (0.5g/mL) changes in the MIC of drug-resistant Salmonella western medicine (μg/mL).

It can be seen from the above table that the minimum inhibitory concentrations of antibacterial drugs against drug-resistant Salmonella are all reduced by more than 4 times, and it can be judged that the herb has a drug resistance reversal effect on drug-resistant Salmonella.

Two, the application example of compound medicine of the present invention

Preparation of Example 1-Example 10 according to the formula in Table 3 and the following method: After crushing the 60-mesh sieve of the grass, as the basic raw material, put it into the mixer according to the weight ratio of the powdered antibacterial drug, and mix evenly. According to conventional methods, the compound medicine is prepared into a preparation product.

The formula of table 3 embodiment 1-10 and preparation dosage form

serial number Antibacterials Pearl grass dosage form Example 1 Amikacin 20g 80g Powder Example 2 Amikacin 13g 10g tablet Example 3 Amikastar 7g 160g oral liquid Example 4 Amikastar 30g 110g Granules Example 5 Amikacin 15g 120g Powder

Example 6 Amikacin 25g 50g tablet Example 7 Amikacin 10g 80g oral liquid Example 8 Amikastar 30g 160g Granules Example 9 Amikacin 3g 5g Powder Example 10 Amikastar 30g 200g tablet

The inventor obtained the compound drug product by the above-mentioned embodiment and carried out the following experiments:

1. The curative effect experiment of the compound drug product of embodiment 1-4 gained to the piglet diarrhea of 40 days of age

Clinical symptoms: In a pig farm in Fangchenggang, Guangxi, 40-day-old piglets began to have diarrhea after weaning and turning around, and pulled watery feces. The feces contained undigested feed. Diarrhea, dehydration and death, no symptoms of vomiting, the farmers used a variety of drugs, but the treatment did not have the desired effect.

Necropsy lesions: After necropsy, the intestinal wall of the pig is thin, especially in the rear end of the small intestine and the colon, the intestinal mucosa is shed, and the intestinal wall is congested and bleeding.

Laboratory Diagnosis: Stained slices of the lesioned liver were taken, Gram stained, observed under a microscope, and red Gram-negative bacilli were seen; it was preliminarily determined to be Escherichia coli disease. Through Escherichia coli culture and biochemical test identification, it was judged that the piglets were infected with Escherichia coli disease, and PCR amplification proved that there were CTX-M type ESBLs drug resistance gene and rosA3 drug resistance gene.

Experimental drug regimen: the diseased pigs were divided into 6 groups for administration, the first group was administered according to the addition amount of 1.5g/kg feed in Example 1, and the second group was administered according to the addition amount of 0.5g/kg feed in Example 2 , the third group is administered according to the addition amount of 7g/kg feed with embodiment 3, and the fourth group is administered according to the addition amount of 1.3g/kg feed with embodiment 4; The fifth group uses amikacin powder according to the dosage of 0.3g The addition amount of /kg feed was administered, and the sixth group was administered with the addition amount of 7g/kg feed; the above six groups of medication regimens were used continuously for 5 days.

Feedback on the use effect: 2 days after the first group to the fourth group of medication, the pigs no longer pull watery feces, the feces are formed, and the feces no longer contain feed. Continue to use for 3 days, and the pigs are cured; 2 days after the fifth group of medication, The pigs are still unhealthy, continue to use for 3 days, and basically recover to health, but some still have mushy feces, but after 3 days of stopping the drug, the pigs relapse; 2 days after the sixth group of drugs, there is no obvious improvement in the pigs, continue to use for 3 days , the pigs continued to have diarrhea, and the rest were more serious. It can be seen that the curative effect of the compound drug containing C. chinensis is better than that of amikacin and C. singulae alone on pigs.

2. The curative effect experiment of the compound drug product obtained in Examples 5-10 on 30-day-old meat duck diarrhea

Clinical symptoms: In a duck farm in Nanning, Guangxi, 30-day-old meat ducks pulled watery feces, and there were obvious undigested feed in the feces. Individual ducks with severe diarrhea gradually appeared soft feet and died slowly.

Necropsy lesions: the dead ducks were seriously dehydrated, some ducks were still quite large, their livers were swollen and congested, some ducks had white membranes on their livers, and the air sacs were cloudy. The initial diagnosis was E. coli or duck serositis.

Experimental drug regimen: the diseased ducks were divided into 8 groups for administration, the first group was administered according to the addition amount of 2.5g/kg feed with Example 5, and the second group was administered with Example 6 according to the addition amount of 0.9g/kg feed , the third group is administered according to the addition amount of 2.5g/kg feed with embodiment 7, and the fourth group is administered according to the addition amount of 1.7g/kg feed with embodiment 8; the fifth group is administered with embodiment 9 according to 0.8g/kg feed The dosage administration of kg feed, the sixth group is administered according to the dosage addition of 2.3g/kg feed with embodiment 10; The seventh group is administered according to the dosage addition of 0.3g/kg feed with amikacin powder, the eighth group The group was administered with 4 g/kg of feed in the coarse powder of Containing Grass; the above-mentioned eight groups of medication schemes were used continuously for 5 days.

Feedback on the use effect: 3 days after the first group to the sixth group of medication, the sick ducks no longer pull watery feces, the feces are formed, and no dead ducks appear. Continue to use for 2 days, and the ducks are basically cured; 3 days after the seventh group of medication, The effect of the duck flock is not obvious, many of them still pull watery feces, containing undigested feed, continue to use for 2 days, a little better, but not completely, dilute watery feces still appear, use the compound drug obtained in Example 5 instead, After continuing to use for 3 days, all the feces became normal; after 3 days of using the eighth group, the ducks did not have much obvious improvement, and continued to use for 2 days, and the death of ducks was still not effectively controlled. It can be seen that the curative effect of the compound medicine containing Zhazhucao on duck diarrhea is better than that of the single medicine.

3. The curative effect experiment of the compound drug product obtained in Example 5-8 on Salmonella chicken disease

Clinical symptoms: In a chicken farm in Liuzhou, Guangxi, 15-day-old broiler chickens had diarrhea, white, pasty and loose feces, the fluff around the anus was contaminated by feces, there were lime-like lumps, and some screamed in pain. death phenomenon. Autopsy of individual chicken liver enlargement is dark red to dark purple, or slightly khaki, crisp and easy to break, the surface is scattered or densely covered with grayish-white, grayish-yellow necrotic spots, sometimes red bleeding spots, typical characteristics of salmonellosis.

Laboratory Diagnosis: Stained slices of the lesioned liver were taken, Gram stained, observed under a microscope, and red Gram-negative bacilli were seen; it was preliminarily judged to be Salmonella disease. Through Salmonella culture and biochemical test identification, it was judged that the chicken infection was Salmonella disease, and the results of the drug susceptibility test were judged to be drug-resistant Salmonella strains according to the NCCLS standard.

Experimental medication scheme: the diseased chickens are divided into 6 groups for administration, the first group is administered according to the addition amount of 2.5g/kg feed with Embodiment 5, and the second group is administered according to the addition amount of 0.9g/kg feed with Embodiment 6. Medicine, the third group uses embodiment 7 according to the dosage administration of 2.5g/kg feed, the 4th group uses embodiment 8 according to the dosage administration of 1.7g/kg feed; The 5th group uses amikacin powder according to The dosage of 0.3g/kg feed was administered, and the sixth group was administered with the addition amount of 4g/kg feed with Grass japonicus coarse powder; the above six groups of medication schemes were used continuously for 5 days.

Effect feedback: After the first group to the fourth group of chickens took medicine, the feed intake of the diseased chickens increased, and no dead chickens appeared. After 3 days of continuous use, the chickens were basically cured; Most of the flocks are still unhealthy, and some chickens still die. Continue to use for 3 days, and the death of chickens has not been effectively controlled; after 2 days of medication, the chickens in the sixth group have not changed significantly. Continue to use for 3 days, the chickens The death phenomenon has not been effectively controlled. It can be seen that the curative effect of the compound medicine containing Zhucao is better than that of the single prescription.

Claims (2)

1. contain the poultry compound medicine containing pearl grass and amikacin, it is characterized in that, by the raw material of following weight portion, formed: amikacin 3-30 part, contains the careless 5-200 part of pearl.

2. the poultry compound medicine containing containing pearl grass and amikacin according to claim 1, is characterized in that, described compound medicine is prepared into powder, tablet, oral liquid or granule.