CN102631474A - Nano emulsion for treating anaerobic infections of intestinal tracts of livestocks and preparation method of nano emulsion - Google Patents
Nano emulsion for treating anaerobic infections of intestinal tracts of livestocks and preparation method of nano emulsion Download PDFInfo
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- CN102631474A CN102631474A CN2012101344040A CN201210134404A CN102631474A CN 102631474 A CN102631474 A CN 102631474A CN 2012101344040 A CN2012101344040 A CN 2012101344040A CN 201210134404 A CN201210134404 A CN 201210134404A CN 102631474 A CN102631474 A CN 102631474A
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Abstract
The invention discloses a nano emulsion for treating anaerobic infections of intestinal tracts of livestocks. The nano emulsion comprises the following raw materials in percent by mass: 0.1-8.0 percent of linaloe oil, 0.1-7.0 percent of carvacrol, 0.01-0.7 percent of metronidazole, 19.0-35.0 percent of surfactant, 0-5.0 percent of cosurfactant and the balance of distilled water, wherein the sum of the mass percent of the raw materials is 100 percent. The nano emulsion has stronger inhibiting and killing functions to most of obligate anaerobes, and is mainly used for treating enteritidis caused by infections of anaerobic bacteria such as bacteroides, clostridium, aerogenesis bacillus, peptococcus, eubacteria and streptococcus faecalis, for the livestocks. The nano emulsion has the advantages of high bioavailability, remarkable antibacterial action and treatment effect, and wide market prospect in the field of veterinary medicines.
Description
Technical field
[0001] the invention belongs to field of veterinary, relate to a kind of medicine that can be used for the enteritis that poultry cause because of anaerobic infection, be specifically related to a kind of nano-emulsion that is used to treat the animal and bird intestines anaerobic infection and preparation method thereof.
Background technology
Anaerobe is extensively to be present in natural its metabolic process not need oxygen to participate in the quasi-microorganism flora that can grow and breed.Mainly comprise gram negative bacilli, gram-positive cocci, Gram-negative coccus etc.Anaerobe is a main flora in the body intestinal, can account for more than 99% of all totals number of bacteria, under the normal condition; Owing to receive mutual restriction and the influence of intestinal environment between the different floras; General be in the Mutualism state with body, body can not fallen ill, but when immunity of organisms lowly or during the intestinal environment change; Anaerobe just can be waited for an opportunity a large amount of breedings, causes bacterial enteritis and health is threatened.
When causing enteritis when poultry generation anaerobic infection, stench feces is drawn in animal performance appetite depression or abolishment, is with blood sometimes in the feces, and the animal performance is become thin, and happiness is crouched and lied prostrate, and is reluctant to walk about, and lethargy usually breaks away from colony.Primary disease is as if untimely treatment, and the state of an illness increases the weight of easily, and is dead because of dehydration at last.Simultaneously, because the intestinal wall mucosa is destroyed, pathogen can wait for an opportunity to get into blood through intestinal wall, causes bacteremia and septicemia and quickens animal dead.Some pathogen has the aerogenesis characteristic, and such as bacillus perfringens etc., the gas of these antibacterial outputs under normal circumstances can be got rid of by the rhythmic wriggling of intestinal; But when intestinal is inflamed; The intestinal peristalsis promoting rule is destroyed on the one hand, causes gas to discharge obstacle, on the other hand because a large amount of breedings of pathogen; Total gas flow of output finally can make animal the intestinal air-blowing occur and death than increasing greatly under the normal condition.
Primary disease has certain infectiousness, mainly through alimentary infection, in the breeding process, in case the symptom that discovery has animal to have has loose bowels quarantines it immediately, feces is cleared up, and environment is carried out thorough disinfection, provides against hidden trouble.The morbidity animal is treated targetedly, and at present commonly used have little ecological therapy and a pharmacotherapy.Little ecological therapy is the exclusiveness of utilizing between microorganism species; Utilize the advantage of probiotic bacteria and suppress the breeding of pathogen, but this method at present technology that is that all right is ripe, especially duodenum of animal and jejunum leading portion; The intestinal juice environment is difficult to be fit to growth of microorganism; General oral microbial ecological agent is when arriving this section, and probiotic bacteria meeting mortality is difficult to effectively arrive the affected part intestinal segment; Pharmacotherapy is the main therapy of present aquaculture, mainly is to use antibiotic, though effectively, caused a lot of social problems and food-safety problem.Too much use antibiotic not only can make aquaculture cost rise, and also can select the drug resistance flora simultaneously, and in the course of time, the drug resistance of flora strengthens gradually, and recovering raises difficult questions controls.Because the system of China is gone back imperfection at present; There is quite a few plant blindly to pursue interests; Do not follow the medicine off-drug period, make medicine pass through food such as meat, egg, milk and get into human body, health is threatened; The meat product of China also repeatedly influences outlet because of antibiotic remains exceeds standard, and has caused a series of social problems.
The pharmaceutical dosage form of treatment poultry anaerobic infection enteritis is main with former powder, powder and injection at present; But former powder and powder non-uniform phenomenon occurs stirring because of consumption less easily when spice; Some former powder drug solubility when the drinking-water administration is low or dissolving is slow, causes situation such as poultry dosage deficiency or obstruction aqueduct.Drug administration by injection is wasted time and energy, and animal stress be big, is unfavorable for the jumpbogroup operation, and is fast not as oral curative effect aspect treatment digestive tract enteritis.Therefore, be necessary to research and develop a kind of novel pharmaceutical formulation and replace conventional dosage forms, improve the deficiency of original dosage form, meet the need of market.
Summary of the invention
The present invention is directed to the defective of the problems referred to above and prior art, provide a kind of bioavailability good, stability is high to be used for the nano-emulsion of the treatment of the enteritis that the animal and bird intestines anaerobic infection causes.This nano-emulsion of experiment proof safely, have no side effect and evident in efficacy.
The technical scheme that realizes the foregoing invention purpose is this nano-emulsion that is used to treat the animal and bird intestines anaerobic infection, is made up of following raw materials by weight percentage:
Linaloe oil 0.1%～8.0%
Carvacrol 0.1%～7.0%
Metronidazole 0.01%～0.7%
Surfactant 19.0%～35.0%
Cosurfactant 0～5.0%
Surplus is a distilled water
The mass percent sum of above-mentioned raw materials is 100%;
On surfactant selection, the present invention selects the nonionic surfactant of avirulence and good biocompatibility for use.Nonionic surfactant is more stable in solution, is not subject to the influence of strong electrolyte, inorganic salts, also is not subject to the influence of soda acid, and good with the compatibility of other surfactants, and haemolysis is less.In theory; The HLB value that the preparation of O/W type nanometer emulsion needs surfactant is between 8～18; Consider preparation technology's simplicity; Be the easy formation property of nano-emulsion and the stability of the nano-emulsion for preparing, the present invention selects the liquid nonionic surfactant of HLB between 10～15 for use.The present invention selects three kinds of tween 80, polyoxyethylene ether Oleum Ricini (EL-40) and polyoxyethylene (40) castor oil hydrogenated (RH-40).
The present invention with linaloe oil, carvacrol and metronidazole as combination; Be because the linaloe oil main component is terpenoids such as caryophyllene oxide, eucalyptol; These chemical compounds are the Lignum Aquilariae Resinatum main component of essential oil, and its content height is directly relevant with drug effect, has effects such as promoting the circulation of QI to relieve pain, warming middle-JIAO to arrest vomiting, helping inspiration to relieve asthma; Can treat abdominal distention, vomiting singultus etc., gastrointestinal tract is had conditioning and health-care effect; Carvacrol has bactericidal activity efficiently, can act on the bacteria cell wall structural protein, makes its degeneration and solidifies, and penetrates the pathogenic microorganism cell membrane rapidly, and cellular content is run off, and destroys that crucial metabolic process finally makes pathogen dead in the cell; Metronidazole is the nitro glyoxaline antibacterials; Its contained nitro reducible one-tenth in oxygen-free environment is amino and show the anaerobe resistant effect; Its antimicrobial spectrum comprises Bacteroides, Clostridium, aerogenesis bacillus cereus, Peptococcus, Eubacterium and streptococcus faecalis etc., has ideal therapeutic effect; After the said medicine combination, no incompatibility, curative effect strengthens.
Cosurfactant of the present invention is not the essential condition that this nanoemulsions forms, and at cosurfactant is can form a kind of nano-emulsion that is used to treat the animal and bird intestines anaerobic infection under zero the situation yet.
The present invention detects through transmission electron microscope, and droplet diameter distribution is between 1～100nm, and outward appearance is colourless or flaxen transparency liquid, has stability preferably:
1, ageing stability
Nano-emulsion of the present invention is at storage under the room temperature nature change condition after 3 months, through the lasting clear of check outward appearance, finds that muddiness, layering, medicine the breakdown of emulsion situation such as separate out, and explains that ageing stability is good.
2, heat storage stability
This nano-emulsion is sub-packed in several vials, places 50 ℃ of calorstats to store 15d after the sealing, the every sampling at a distance from 5d observed.The result shows, this nano-emulsion heat storage back check outward appearance still keeps the outward appearance of clear, finds that muddiness, layering, medicine the breakdown of emulsion situation such as separate out, and explains that heat storage stability is good.
3, anti-freeze-stable property
With this nano-emulsion in refrigerator-10 ℃ preserve a week after.Return to room temperature.This nano-emulsion can be liquid by solid-state thawing, and outward appearance clear is still placed all continued and checked through dilution check back, and discovery muddiness, layering, medicine such as separate out at the breakdown of emulsion situation, show that freezing-resistance is good.
4, accelerated stability
This nano-emulsion is placed test tube, sealing, under the rotating speed of 15000r/min centrifugal 20 minutes, outward appearance clear still after the dilution check, discovery muddiness, layering, medicine such as separate out at the breakdown of emulsion situation.
5, light stability experiment
This nano-emulsion of part is loaded in the vial, and sealing places under the daylight, and light at room temperature is taken a sample respectively in 1d, 3d, 5d, 10d according to 10d.Keep clear through this nanoemulsions of dilution inspection show, do not have muddiness, layering, medicine and the breakdown of emulsion situation such as separate out, show that light stability is good.
A kind of nano-emulsion that is used to treat the animal and bird intestines anaerobic infection of the present invention has the effect of the enteritis that treatment causes because of the intestinal anaerobic infection.
Usage: oral or drinking-water.
A kind of nano-emulsion that is used to treat the animal and bird intestines anaerobic infection of the present invention compared with prior art has the following advantages:
1) good water solubility.Can be in water infinite dilution and can breakdown of emulsion, during use dissolving evenly, blocking pipe not, the animal-use drug amount is neat;
2) thermodynamic stability is high.This nano emulsion composition stores not phase-splitting, does not precipitate between-10 ℃ and 50 ℃, not muddy, the storage time is long, and stability improves;
3) bioavailability is high.This nano-emulsion emulsion droplet particle diameter is easy to get in the pathogen thalline through cell membrane and brings into play curative effect between 1 ~ 100nm; Compare with general formulation, reduced drug dose, shortened the off-drug period greatly, medicine noresidue or less residue;
4) improved the dissolubility of metronidazole in water;
5) method technology is simple, is fit to large-scale production.
The specific embodiment
Below come the further safety and the beneficial effect of elaboration medicine according to the invention through drug safety test with the comparative efficacy test.
Test Example 1Nanoemulsion medicine safety testing of the present invention
This experiment adopts the improvement karber's method to survey the LD of animal
50, laboratory animal is 1 monthly age laying chickling, The time limit of experiment was two weeks.
With 70 one the monthly age health-care eggs chickling be divided into 7 groups at random; Every group 10; Wherein first group is the experiment contrast group; Second group is that nano-emulsion stock solution group of the present invention, the 3rd group are the nano-emulsion stock solution of 0.6 times of dilution, and the like, concentration of each group medicinal liquid is 0.6 times of last group of concentration (group that is the design of this acute toxicity testing is apart from being 0.6).Each treated animal is weighed respectively; Each group is irritated the corresponding nanoemulsions of clothes according to the dosage of 10g/kg body weight respectively, and matched group is irritated the normal saline of clothes equivalent, and each treated animal was raised in same environment following 14 days after the dispenser; Observe the mental status, diet situation and the active situation etc. of animal during this time; After finishing, experiment, observes each organ lesion situation with dead sacrifice of animal, and record.
The result shows: each organizes chicken crowd active situation and health status is all normal, cuts open the also no abnormal pathological changes of inspection each internal organs of back, zero difference between each group, LD
50﹥ 10g/kg body weight, the nontoxic scope in true border, this result show that nanoemulsions safety of the present invention is good.
Test Example 2Nano-emulsion of the present invention and the test of metronidazole treatment rabbit clostridium property diarrhoea effect comparison
Clostridium property diarrhoea is that the common alimentary infection of rabbit is sick, and pathogen is a bacillus perfringens, and primary disease is multiple when change in weather; Mostly morbidity rabbit crowd is the young rabbit below 4 monthly ages, and symptom mainly shows as lethargy, inappetence, the soft distortion of loose and watery stool or is gelatin, is with blood sometimes; Pathogen will breed in a large number in the rabbit intestinal if untimely treatment is suffered from, and constantly aerogenesis causes abdominal part to expand, and has knocked tympanitic resonance; Body weight falls sharply, and blows hair and observes skin purple darkly, gets into the generally very difficult treatment of this phase.
Choose and face 18 of the trouble rabbits of examining the natural occurrence clostridium property diarrhoea initial stage, body weight is divided into 3 groups of matched group, Metronidazole group and this nano-emulsion groups, 6 every group with it at random between 1.9 ~ 2.1kg.Every of matched group is irritated clothes normal saline 10ml; Every oral metronidazole 50mg of Metronidazole group, this nano-emulsion group are irritated clothes this nanoemulsions of 10g (drug loading is 0.5%), once a day, and continuous use 3 days; Observe during this time and suffer from rabbit diet, activity and death condition, and note down.
Curative effect judging standard:
Invalid: as to suffer from that the rabbit abdominal part expands, body weight falls sharply, not improvement of appetite, sx or death;
Produce effects: trouble rabbit appetite is improved, feces is shaped, spirit is good, the behavior recovery;
Cure: suffer from rabbit feces and be shaped behavior performance, diet situation and active situation and healthy rabbit indifference;
Total effective rate=obvious effective rate+cure rate
Table 1This nano-emulsion and the test of metronidazole treatment rabbit clostridium property diarrhoea effect comparison
Can be known by table 1: all suffer from rabbit death full matched group, and the Metronidazole group cure rate is 33%, and obvious effective rate is 17%, and total effective rate is 50%; This nano-emulsion group cure rate is 67%, and obvious effective rate is 0%, and total effective rate is 67%.
Can be found out by the result: the nano-emulsion treatment rabbit clostridium property diarrhoea effect that the present invention is a kind of to be used to treat the animal and bird intestines anaerobic infection is superior to the former medicine of metronidazole.
Below come further to set forth medicine according to the invention through specific embodiment composition and method for preparing.
Embodiment 1
1) accurately take by weighing linaloe oil 3.0g, carvacrol 3.0g, metronidazole 0.3g, tween 80 7.0g, polyoxyethylene ether Oleum Ricini (EL-40) 20.0g, 1 by prescription, 2-propylene glycol 2.0g, distilled water 64.7g, subsequent use;
2) metronidazole with said amount is dissolved in the carvacrol of said amount, and adds the linaloe oil of said amount, and the back that stirs is as oil phase;
3) surfactant and the cosurfactant with said amount joins step 2) preparation oil phase in, stir fast;
4) distilled water is slowly joined as water in the mixture of step 3) preparation, the limit edged stirs, and this system can occur becoming sticky thickly again by the thinning process of thickness by rare, and final system clear is nanoemulsions of the present invention.
Embodiment 2
Form by linaloe oil 8.0g, carvacrol 0.1g, metronidazole 0.01g, polyoxyethylene ether Oleum Ricini (EL-40) 32.0g, dehydrated alcohol 1.0g, distilled water 58.89.
Embodiment 3
By linaloe oil 2.0g, carvacrol 4.0g, metronidazole 0.3g, polyoxyethylene (40) castor oil hydrogenated (RH-40) 23.0g, polyoxyethylene ether Oleum Ricini (EL-40) 5.0g, 1,2-propylene glycol 3.0g, distilled water 62.7g form.
Embodiment 4
By linaloe oil 4.0g, carvacrol 2.0g, metronidazole 0.1g, polyoxyethylene (40) castor oil hydrogenated (RH-40) 25.0g, tween 80 5.0g, 1,2-propylene glycol 1.0g, distilled water 62.9g form.
Embodiment 5
Form by linaloe oil 6.0g, carvacrol 1.0g, metronidazole 0.05g, tween 80 12.0g, polyoxyethylene ether Oleum Ricini (EL-40) 20.0g, distilled water 60.95g.
Embodiment 6
Form by linaloe oil 0.1g, carvacrol 7.0g, metronidazole 0.7g, polyoxyethylene (40) castor oil hydrogenated (RH-40) 5.0g, polyoxyethylene ether Oleum Ricini (EL-40) 22.0g, dehydrated alcohol 5.0g, distilled water 55.9g.
Embodiment 7
By linaloe oil 1.5g, carvacrol 2.5g, metronidazole 0.2g, polyoxyethylene ether Oleum Ricini (EL-40) 30.0g, tween 80 5.0g, 1,2-propylene glycol 0.5g, distilled water 60.3g form.
Embodiment 8
Form by linaloe oil 7.0g, carvacrol 1.0g, metronidazole 0.05g, polyoxyethylene ether Oleum Ricini (EL-40) 31.0g, distilled water 60.95g.
Embodiment 9
Form by linaloe oil 2.0g, carvacrol 2.0g, metronidazole 0.1g, tween 80 24.0g, dehydrated alcohol 3.0g, distilled water 68.9g.
Embodiment 10
Form by linaloe oil 2.5g, carvacrol 5.0g, metronidazole 0.5g, polyoxyethylene (40) castor oil hydrogenated (RH-40) 9.0g, tween 80 10.0g, dehydrated alcohol 4.0g, distilled water 69.0g.
Claims (3)
1. a nano-emulsion that is used to treat the animal and bird intestines anaerobic infection is characterized in that, is made up of following raw materials by weight percentage:
Linaloe oil 0.1%～8.0%, carvacrol 0.1%～7.0%, metronidazole 0.01%～0.7%, surfactant 19.0%～35.0%, cosurfactant 0～5.0%, surplus are distilled water, and the mass percent sum of above-mentioned raw materials is 100%;
One or more mixture in described glass or plastic containers tween 80, polyoxyethylene ether Oleum Ricini and the polyoxyl 40 hydrogenated castor oil;
Described cosurfactant is dehydrated alcohol and 1, any one in the 2-propylene glycol.
2. a kind of nano-emulsion that is used to treat the animal and bird intestines anaerobic infection according to claim 1 is characterized in that the particle diameter of this nano-emulsion is between 1～100nm.
3. a kind of nano-emulsion method for preparing that is used to treat the animal and bird intestines anaerobic infection according to claim 1 is characterized in that, may further comprise the steps:
1) accurately take by weighing linaloe oil, carvacrol, metronidazole, surfactant, cosurfactant and distilled water by prescription, subsequent use;
2) metronidazole with said amount is dissolved in the carvacrol of said amount, and adds the linaloe oil of said amount, and the back that stirs is as oil phase;
3) surfactant and the cosurfactant with said amount joins step 2) preparation oil phase in, stir fast;
4) distilled water is slowly joined as water in the mixture of step 3) preparation, the limit edged stirs, and this system can occur becoming sticky thickly again by the thinning process of thickness by rare, and final system clear promptly gets.
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| CN106343163A (en) * | 2016-08-29 | 2017-01-25 | 厦门惠盈动物科技有限公司 | Emulsified nano oregano oil and garlic oil additive and preparation method thereof |
| CN107197860A (en) * | 2016-03-17 | 2017-09-26 | 新加坡国立大学 | Nano-emulsion containing carvacrol and preparation method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107197860A (en) * | 2016-03-17 | 2017-09-26 | 新加坡国立大学 | Nano-emulsion containing carvacrol and preparation method thereof |
| CN107197860B (en) * | 2016-03-17 | 2021-12-07 | 新加坡国立大学 | Nanometer emulsion containing carvacrol and preparation method thereof |
| CN106343163A (en) * | 2016-08-29 | 2017-01-25 | 厦门惠盈动物科技有限公司 | Emulsified nano oregano oil and garlic oil additive and preparation method thereof |
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Application publication date: 20120815 |