CN102580093B - Injection for preventing piglet diseases and preparation method thereof - Google Patents
Injection for preventing piglet diseases and preparation method thereof Download PDFInfo
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- CN102580093B CN102580093B CN2011104582205A CN201110458220A CN102580093B CN 102580093 B CN102580093 B CN 102580093B CN 2011104582205 A CN2011104582205 A CN 2011104582205A CN 201110458220 A CN201110458220 A CN 201110458220A CN 102580093 B CN102580093 B CN 102580093B
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Abstract
The invention discloses an injection for preventing piglet diseases. The injection is prepared from the following raw materials: 100-120mg of ovomucoid, 20-50mg of antibiotics, 1-2ml of buffer solution and 1.5-3.5ml of water for injection. Due to the adoption of the ovomucoid, the discharging time of lincomycin hydrochloride can be delayed and prolonged, the blood concentration of lincomycin hydrochloride is increased, the acting time of lincomycin hydrochloride is prolonged, and the effect of realizing a good curative effect with a small dosage is achieved.
Description
Technical field
The present invention relates to a kind of injection and preparation method thereof, particularly relate to a kind of injection that prevents the piglet disease and preparation method thereof.
Background technology
At present, it is many that the pig-breeding production of China is being faced with swine diseases, difficult control, dead high, medication is big, high investment, the predicament of low repayment, the breed unit all can suffer from any time because of losing to defeat and come to an end, people are in order to prevent the various diseases of piglet, the pleura pneumonia that causes of positive bacteria for example, streptococcicosis, staphylococcus aureus etc., a large amount of antibiotic and all feeds additives of using, this and current its people disagree the strong request and the cry of carnivorous foodsafety, again because the pathogenic microorganism of swine diseases, because of contacting with medicine for a long time, from drug resistance to the transition of variability direction, cause existing antibiotic to be difficult to reach again effective preventive effect.
Summary of the invention
The present invention seeks in order to solve the deficiency of above-mentioned technology, a kind of injection that prevents the piglet disease and preparation method thereof is provided, the present invention utilizes ovum gallinaceum mucoprotein as carrier, slow release and the efflux time that has prolonged lincomycin hydrochloride, the blood drug level and the action time that have strengthened lincomycin hydrochloride.
In order to achieve the above object, technical scheme provided by the present invention is: a kind of injection that prevents the piglet disease, it comprises that following raw material makes: the mucoprotein 100mg – of ovum gallinaceum 120mg, antibiotic 20mg-50mg, buffer 1ml-2ml and water for injection 1.5ml-3.5ml.
Further, described buffer is the phosphate buffer of PH7-7.2.
Further, described antibiotic is a lincomycin hydrochloride.
Further, described antibiotic is a gentamycin sulfate.
A kind of preparation method of preventing the injection of piglet disease, it comprises the steps:
(1) get an amount of water for injection with ovum gallinaceum is mucoprotein be diluted to the mucoprotein solution of ovum gallinaceum after, mix with buffer, slowly stirred 15-35 minute;
(2) get an amount of water for injection with after the antibiotic dissolving, mixes with above-mentioned mixed liquor, and slowly stirred 3 – 7 minutes, promptly get the described injection that prevents the piglet disease.
Further, described buffer is 1ml.
Further, the volume of the antibiotic solution of making in step (2) is 1.2ml.
Further, the injection of described prevention piglet disease is 4ml.
Adopt technique scheme, technique effect of the present invention has:
(1) the present invention is mucoprotein as carrier with ovum gallinaceum, by artificial immunity, stimulates pig body generation humoral immunization and cellular immunization, thereby utilizes the immunity of self to kill the intravital target cell of intrusion.
(2) but the present invention utilize the mucoprotein slow release of ovum gallinaceum and prolonged the efflux time of lincomycin hydrochloride, the blood drug level and the action time that have strengthened lincomycin hydrochloride, realize the effect of the big curative effect of low dose of performance.
The specific embodiment
The mucoprotein antigenicity that can utilize self in the present invention of ovum gallinaceum, stimulate the pig body, improve the immune antibody of pig body self, strengthen the ability of self resisting disease, and owing to ovum gallinaceum is mucoprotein is a kind of colloid thing, but the efflux time of slow release and prolongation lincomycin hydrochloride, is realized the effect of the big curative effect of low dose of performance at the blood drug level and the action time that have strengthened lincomycin hydrochloride.
Lincomycin hydrochloride is as narrow-spectrum antibiotic, gram positive coccus there is better effect, especially anaerobic bacteria, golden Portugal bacterium and streptococcus pneumoniae are had efficiently, its mechanism of action is similar with erythromycin, belong to antibacterial, be mainly used in responsive microbial various infection, as pneumonia, meningitis, endocarditis, cellulitis, tonsillitis, erysipelas, furuncle and urinary system infection etc.
Gentamycin sulfate all has antibacterial and bactericidal action as the aminoglycosides broad ectrum antibiotic to multiple gram-negative bacteria and positive bacteria.Stronger to effects such as gram-negative bacteria such as bacillus pyocyaneus, aerobacteria, pneumobacillus, Salmonella, escherichia coli and Bacillus proteus and golden Portugal bacterium.
But buffer is the Balancing degree in the present invention, avoids injecting porkling of the present invention and stress occurs.
Water for injection is regulated the concentration of each component in the present invention as a kind of solvent, helps porkling and absorbs.
Below being embodiment provided by the present invention, only is to further specify application of the present invention, rather than limits.
Embodiment 1
A kind of injection that prevents the piglet disease, it comprises that following raw material makes:
The mucoprotein 120mg of ovum gallinaceum;
Lincomycin hydrochloride 24mg;
Buffer 1ml;
Water for injection 3ml;
Wherein, buffer is the phosphate buffer of PH7.2.
A kind of preparation method of preventing the injection of piglet disease, it comprises the steps:
(1) get 1.2ml water for injection with the 120mg ovum gallinaceum is mucoprotein be diluted to the mucoprotein solution of ovum gallinaceum after, mix with the 1ml buffer, slowly stirred 30 minutes, mixed liquor produces bubble when preventing to stir;
(2) get 1.2ml water for injection with after the 24mg lincomycin hydrochloride dissolving, mixes with above-mentioned mixed liquor, and slowly stirred 5 minutes, reuse 0.6ml water for injection dilutes, and promptly gets the described injection that prevents the piglet disease.
Embodiment 2
A kind of injection that prevents the piglet disease, it comprises that following raw material makes:
The mucoprotein 120mg of ovum gallinaceum;
Gentamycin sulfate 48mg;
Buffer 1ml;
Water for injection 3ml;
Wherein, buffer is the phosphate buffer of PH7.2.
A kind of preparation method of preventing the injection of piglet disease, it comprises the steps:
(1) get 1.2ml water for injection with the 120mg ovum gallinaceum is mucoprotein be diluted to the mucoprotein solution of ovum gallinaceum after, mix with the 1ml buffer, slowly stirred 30 minutes;
(2) get 1.2ml water for injection with after the 48mg gentamycin sulfate dissolving, mixes with above-mentioned mixed liquor, and slowly stirred 5 minutes, reuse 0.6ml water for injection dilutes, and promptly gets the described injection that prevents the piglet disease.
Embodiment 3
A kind of injection that prevents the piglet disease, it comprises that following raw material makes:
The mucoprotein 100mg of ovum gallinaceum;
Lincomycin hydrochloride 20mg;
Buffer 2ml;
Water for injection 1.5ml;
Wherein, buffer is the phosphate buffer of ph7.
A kind of preparation method of preventing the injection of piglet disease, it comprises the steps:
(1) get 0.8ml water for injection with the 100mg ovum gallinaceum is mucoprotein be diluted to the mucoprotein solution of ovum gallinaceum after, mix with the 2ml buffer, slowly stirred 15 minutes;
(2) get 0.7ml water for injection with after the 20mg lincomycin hydrochloride dissolving, mixes with above-mentioned mixed liquor, and slowly stirred 3 minutes, promptly get the described injection that prevents the piglet disease.
Embodiment 4
A kind of injection that prevents the piglet disease, it comprises that following raw material makes:
The mucoprotein 130mg of ovum gallinaceum;
Gentamycin sulfate 40mg;
Buffer 2ml;
Water for injection 3.5ml;
Wherein, buffer is the phosphate buffer of ph7.
A kind of preparation method of preventing the injection of piglet disease, it comprises the steps:
(1) get 1.5ml water for injection with the 130mg ovum gallinaceum is mucoprotein be diluted to the mucoprotein solution of ovum gallinaceum after, mix with the 2ml buffer, slowly stirred 35 minutes;
(2) get 2ml water for injection with after the 40mg gentamycin sulfate dissolving, mixes with above-mentioned mixed liquor, and slowly stirred 7 minutes, promptly get the described injection that prevents the piglet disease.
Below be the mucoprotein aminoacid sequence of ovum gallinaceum among the present invention:
GACATACAGCTAGAAAGCTGTATTGCCTTTAGCACTCAAGCTCAAAAGACAACTCAGAGTTCACCATGGGCTCCATCGGCGCAGCAAGCATGGAATTTTGTTTTGATGTATTCAAGGAGCTCAAAGTCCACCATGCCAAT
M G S I G A A S M E F C F D V F K E L K V H H A N
GAGAACATCTTCTACTGCCCCATTGCCATCATGTCAGCTCTAGCCATGGTATACCTGGGTGCAAAAGACAGCACCAGGACACAGATAAATAAGGTTGTTCGCTTTGATAAACTTCCAGGATTCGGAGACAGTATTGAAGC
E N I F Y C P I A I M S A L A M V Y L G A K D S T R T Q I N K V V R F D K L P G F G D S I E A
TCAGTGTGGCACATCTGTAAACGTTCACTCTTCACTTAGAGACATCCTCAACCAAATCACCAAACCAAATGATGTTTATTCGTTCAGCCTTGCCAGTAGACTTTATGCTGAAGAGAGATACCCAATCCTGCCAGAATACT
Q C G T S V N V H S S L R D I L N Q I T K P N D V Y S F S L A S R L Y A E E R Y P I L P E Y
TGCAGTGTGTGAAGGAACTGTATAGAGGAGGCTTGGAACCTATCAACTTTCAAACAGCTGCAGATCAAGCCAGAGAGCTCATCAATTCCTGGGTAGAAAGTCAGACAAATGGAATTATCAGAAATGTCCTTCAGCCAAGC
L Q C V K E L Y R G G L E P I N F Q T A A D Q A R E L I N S W V E S Q T N G I I R N V L Q P S
TCCGTGGATTCTCAAACTGCAATGGTTCTGGTTAATGCCATTGTCTTCAAAGGACTGTGGGAGAAAACATTTAAGGATGAAGACACACAAGCAATGCCTTTCAGAGTGACTGAGCAAGAAAGCAAACCTGTGCAGATGAT
S V D S Q T A M V L V N A I V F K G L W E K T F K D E D T Q A M P F R V T E Q E S K P V Q M M
GTACCAGATTGGTTTATTTAGAGTGGCATCAATGGCTTCTGAGAAAATGAAGATCCTGGAGCTTCCATTTGCCAGTGGGACAATGAGCATGTTGGTGCTGTTGCCTGATGAAGTCTCAGGCCTTGAGCAGCTTGAGAGTA
Y Q I G L F R V A S M A S E K M K I L E L P F A S G T M S M L V L L P D E V S G L E Q L E S
TAATCAACTTTGAAAAACTGACTGAATGGACCAGTTCTAATGTTATGGAAGAGAGGAAGATCAAAGTGTACTTACCTCGCATGAAGATGGAGGAAAAATACAACCTCACATCTGTCTTAATGGCTATGGGCATTACTGAC
I I N F E K L T E W T S S N V M E E R K I K V Y L P R M K M E E K Y N L T S V L M A M G I T D
GTGTTTAGCTCTTCAGCCAATCTGTCTGGCATCTCCTCAGCAGAGAGCCTGAAGATATCTCAAGCTGTCCATGCAGCACATGCAGAAATCAATGAAGCAGGCAGAGAGGTGGTAGGGTCAGCAGAGGCTGGAGTGGATGC
V F S S S A N L S G I S S A E S L K I S Q A V H A A H A E I N E A G R E V V G S A E A G V D A
TGCAAGCGTCTCTGAAGAATTTAGGGCTGACCATCCATTCCTCTTCTGTATCAAGCACATCGCAACCAACGCCGTTCTCTTCTTTGGCAGATGTGTTTCCCCTTAAAAAGAAGAAAGCTGAAAAACTCTGTCCCTTCCAA
A S V S E E F R A D H P F L F C I K H I A T N A V L F F G R C V S P
CAAGACCCAGAGCACTGTAGTATCAGGGGTAAAATGAAAAGTATGTTCTCTGCTGCATCCAGACTTCATAAAAGCTGGAGCTTAATCTAGAAAAAAAATCAGAAAGAAATTACACTGTGAGAACAGGTGCAATTCACTTT
TCCTTTACACAGAGTAATACTGGTAACTCATGGATGAAGGCTTAAGGGAATGAAATTGGACTCACAGTACTGAGTCATCACACTGAAAAATGCAACCTGATACATCAGCAGAAGGTTTATGGGGGAAAAATGCAGCCTTC
CAATTAAGCCAGATATCTGTATGACCAAGCTGCTCCAGAATTAGTCACTCAAAATCTCTCAGATTAAATTATCAACTGTCACCAACCATTCCTATGCTGACAAGGCAATTGCTTGTTCTCTGTGTTCCTGATACTACAAG
GCTCTTCCTGACTTCCTAAAGATGCATTATAAAAATCTTATAATTCACATTTCTCCCTAAACTTTGACTCAATCATGGTATGTTGGCAAATATGGTATATTACTATTCAAATTGTTTTCCTTGTACCCATATGTAATGGG
TCTTGTGAATGTGCTCTTTTGTTCCTTTAATCATAATAAAAACATGTTTAAGC
Below illustrate the mucoprotein preparation method of ovum gallinaceum among the present invention:
1, employing method: organic solvent precipitation method, through DEAE---the cellulose chromatography purification obtains qualified products.
2, reagent and equipment:
(1) propanol (analytical pure);
(2) 0.5mol/L trichloroacetic acid solution;
(3) 0.02mol/LPH6.5 phosphate buffer;
(4) 0.3mol/L NaCL-0.02mol/LPH6.5 phosphate buffer;
(5) 0.5mol/L NaCL-0.02mol/LPH6.5 phosphate buffer;
(6) DEAE---cellulose (DE-32 or 52);
(7) chromatographic column: 2.8 * 30cm;
3, operation:
(1) preparation process
With carrying out the negative egg of bird flu Detection of antigen, under gnotobasis, collect its Ovum Gallus domesticus album 300g, place beaker, beaker is put into 30 ℃ of water-baths, under constantly stirring, add isopyknic trichloroacetic acid---propanol solution (40ml0.5mol/L trichloroacetic acid and 80ml propanol mix, i.e. 1:2V/V).Continue to stir 30min again, be placed on then leave standstill 10-16h under 4 ℃ the condition after, cross its light yellow limpid filtrate of leaching.Get 1ml filtrate and put into water-bath in test tube, under 80 ℃ of slow stirrings, add the propanol 500ml of 4 ℃ of pre-coolings, post precipitation occurs, leave standstill 2h under 4 ℃ condition, sucking filtration obtains the ovomucoid filter cake.
(2) purification of ovomucoid
The ovomucoid filter cake that makes is dissolved in the distilled water, and transferring pH value is 4.5, and the bag filter of packing into is used distill water dialysis, removes trichloroacetic acid, reuse 0.02mol/LPH6.5 phosphate buffer dialysis equilibrium (under 4 ℃ condition); The DEAE that will handle well---cellulose ion exchanger soaked 1-5 minute with the 0.02mol/LPH6.5 phosphate buffer again, after changing clothes 3-6 time, the chromatographic column (2.8 * 30cm) of packing into, continue the chromatographic column of flowing through with the 0.02mol/LPH6.5 phosphate buffer, identical up to the PH of effluent with the buffer solution ph 6.5 that enters.
The ovomucoid upper strata suction post that to dialyse good then with 0.02mol/LPH6.5 phosphate buffer eluting, first peak occurs earlier on detector, this is a foreign protein,
Then use the 5mol/LNaCL-0.02mol/LPH6.5 phosphate buffer instead and soak eluting, second peak appears in this moment, is ovomucoid, uses 0.5mol/L NaCL-0.02mol/LPH6.5 phosphate buffer at last, is eluted to no albumen and occurs.
Merge ovomucoid and collect liquid, add the ammonium sulfate powder, make saturatedly to reach 0.8, behind the 10-16h that saltouts, centrifuging and taking gets precipitate, and it is mucoprotein to be ovum gallinaceum.
The present invention prevents the injection of piglet disease to be mainly used in 5 and the piggy injection of 22 ages in days, clinical trial proves, the mucoprotein injection that is mixed and made into of gentamycin sulfate and ovum gallinaceum is used for the piglet of 5 ages in days, the diseases such as bacillus pyocyaneus, aerobacteria, pneumobacillus, Salmonella, escherichia coli and Bacillus proteus that can effectively prevent positive bacteria to cause; The mucoprotein injection that is mixed and made into of lincomycin hydrochloride and ovum gallinaceum is used for the piglet of 22 ages in days, pneumonia, meningitis, endocarditis, cellulitis, tonsillitis, erysipelas, furuncle and the urinary system infection etc. that can effectively prevent negative bacteria to cause; Can make the sickness rate of piglet drop at ordinary times 50%, embody really: piglet is vivaciously healthy, anosis in good health.
Usage: 1 gentamycin sulfate of injection and the mucoprotein injection that is mixed and made into of ovum gallinaceum during porkling 5 ages in days, 1 lincomycin hydrochloride of injection and the mucoprotein injection that is mixed and made into of ovum gallinaceum when porkling 22 ages in days.
Following table for the present invention to injecting piglet intramuscular injection efficacy experiment of the present invention:
Experimental result:
| Dead (head) | Eliminate on (head) | The examination end survives (head) | Examination phase weightening finish (kg) | Examination phase medication (unit) | |
| Experimental group | Do not have | 1 | 61 | 27.58 | 8.2 unit |
| Matched group | 2 | 1 | 53 | 26.04 | 15.2 unit |
As seen from the above table, experimental group does not have extremely, eliminates 1, accounts for 62 1.16%; Dead 2 of matched group is eliminated 1, accounts for 56 5.4%, differs 3.79%, and difference is fairly obvious.
The ovum gallinaceum mucin is a complete antigen, has two kinds of performances.The one, stimulate body to produce immunoreactive immunogenicity or title antigenicity: the 2nd, with the bonded reactionogenicity of corresponding immune material generation specificity.
As everyone knows, gentamycin sulfate and lincomycin hydrochloride all are molecular weight very low (lincomycin are 461.02), only have the haptenic chemical drugs of reactionogenicity.So, be injected into body separately after very fast (4-6h) will be excreted, the time of keeping drug effect can be very short.With the gentamycin sulfate is example, belongs to aminoglycoside.Antibacterial very easily produces drug resistance to this medicine, for this reason, must accomplish 3-4 administration every day when using this medicine separately, and dosage is wanted " jump ".
But with the ovum gallinaceum mucin compound after, also just had antigenicity, the ovum gallinaceum mucin belongs to the gluey body of macromolecule in addition, after same gentamycin sulfate, the lincomycin hydrochloride combination above two kinds of chemical medicine can be discharged from for a long time under ovum gallinaceum mucin (jelly) effect, make blood drug level keep the long period (2-4 days) in vivo, this has just overcome the drawback of independent use chemical medicine.Experimental result shows: this mixture intramuscular injection effect once is equal to single curative effect with gentamycin 5-8 time fully.
The ovum gallinaceum mucin mixes the back respectively from (pharmacokinetics) angle with gentamycin sulfate, hydrochloric acid woods Mycosporin, can play " mark, this " double effect of controlling.According to being: the ovum gallinaceum mucin can go to control " basis " from the autoimmunity that antigenic angle goes to strengthen the pig body, but since antigen in vivo mechanism need the time, in autoantibody forms as yet, with the disinthibite therapeutical effect of idle period of gentamycin sulfate and hydrochloric acid woods Mycosporin.The two again can be complementary on the contrary, mutually promotes, and can reach single target that can't realize with a certain side at last.
By test of many times, selected birth 5 ages in days (120h) intramuscular injection the present invention is the most reasonable.This be because: too early piglet is to the mucinous immunoreation ability of ovum gallinaceum; Too late, create condition can for again the colibacillary invasion of pathogenicity, lose meaning.If after injection this product, a pig autoantibody will form at the 10-12 age in days, can have gentamycin sulfate to go to get ahead of others but can have during this time when birth 5 ages in days.
Injected the present invention during by piglet 5 ages in days, generated and the digestive tract disease in prevention and control piglet stage is certain to play very big protective effect for the high titre of autoantibody.But we should not forget: need 5-7d could produce corresponding antibody gradually after the ovum gallinaceum mucin is injected into human body for the first time, and titre is also lower, tire time of keeping is also shorter, but owing to stimulate fashion that fraction T cell and the differentiation of B cell Halfway Stopping are arranged for the first time, antigenic information is stored in the born of the same parents, be converted into immunological memory cell, this class cell can be survived the several months to the several years.After stimulated by same antibody, proliferation and differentiation quickens immunoreation rapidly.The titre that produces antibody can be primary 50-100 doubly, and the time of keeping can up to the several months to the several years.Here it is at 5 ages in days with the theoretical foundation of reuse the present invention injection during 17d to 22 age in days at interval after the intramuscular injection of the present invention.
It should be noted that at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although the present invention is had been described in detail with reference to preferred embodiment, those of ordinary skill in the art is to be understood that, can make amendment or be equal to replacement technical scheme of the present invention, and not breaking away from the spirit and scope of technical solution of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (6)
1. injection that prevents the piglet disease, it is characterized in that: it comprises that following raw material makes: the mucoprotein 100mg – of ovum gallinaceum 120mg, antibiotic 20mg-50mg, buffer 1ml-2ml and water for injection 1.5ml-3.5ml, described antibiotic is lincomycin hydrochloride or gentamycin sulfate.
2. the injection of prevention piglet disease according to claim 1, it is characterized in that: described buffer is the phosphate buffer of pH7-7.2.
3. the preparation method of the injection of prevention piglet disease according to claim 1, it is characterized in that: it comprises the steps:
(1) get an amount of water for injection with ovum gallinaceum is mucoprotein be diluted to the mucoprotein solution of ovum gallinaceum after, mix with buffer, slowly stirred 15-35 minute;
(2) get an amount of water for injection with after the antibiotic dissolving, mixes with above-mentioned mixed liquor, and slowly stirred 3 – 7 minutes, promptly get the described injection that prevents the piglet disease.
4. the preparation method of the injection of prevention piglet disease according to claim 3, it is characterized in that: described buffer is 1ml.
5. according to claim 3 or 4 described preparation methoies of preventing the injection of piglet diseases, it is characterized in that: the volume of the antibiotic solution of making in step (2) is 1.2ml.
6. the preparation method of the injection of prevention piglet disease according to claim 5 is characterized in that: the injection of described prevention piglet disease is 4ml.
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| CN105030665B (en) * | 2015-07-24 | 2018-06-29 | 浙江大飞龙动物保健品有限公司 | A kind of long-acting veterinary lincomycin hydrochloride injection and preparation method thereof |
| CN107260663A (en) * | 2017-07-01 | 2017-10-20 | 山东中牧兽药有限公司 | A kind of lincomycin hydrochloride injection and preparation method thereof |
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| CN101708330A (en) * | 2009-08-04 | 2010-05-19 | 胡海玉 | Antibody-containing chicken egg protein injection for controlling swine fever (diseases) |
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