阅读说明：本技术 一种信息素组合物喷雾制剂及其制备方法和应用 (Pheromone composition spray preparation and preparation method and application thereof ) 是由 岑桂英 钱星宇 李伟 吴春山 原敏 何玲飞 于 2019-10-14 设计创作，主要内容包括：本发明涉及信息素组合物,尤其涉及一种信息素组合物喷雾制剂及其制备方法和应用。一种信息素组合物喷雾制剂,该喷雾制剂按质量比计包括：雄甾烯酮和/或雄甾烯醇0.001-0.02%；淫羊藿提取物0.001-0.02%；二甲基亚砜0.2-20%；吐温-800.01-0.5%；防腐剂0.1-1.0%；纯化水80.0-99.0%。该喷雾制剂更为安全可靠,并且可靠地改善母猪的生殖表现(即更高的可配发情率),提高母猪利用率,提高母猪生育力。(The invention relates to a pheromone composition, in particular to a pheromone composition spray preparation, a preparation method and application thereof. A pheromone composition spray preparation, which comprises the following components in percentage by mass: androstenone and/or androstenol 0.001-0.02%; herba Epimedii extract 0.001-0.02%; 0.2 to 20 percent of dimethyl sulfoxide; tween-800.01-0.5%; 0.1 to 1.0 percent of preservative; 80.0-99.0% of purified water. The spray preparation is safer and more reliable, reliably improves the reproductive performance of sows (namely higher matchable estrus rate), improves the utilization rate of the sows and improves the fertility of the sows.)

1. A pheromone composition spray preparation is characterized by comprising the following components in percentage by mass:

androstenone and/or androstenol 0.001-0.02%;

herba Epimedii extract 0.001-0.02%;

0.2 to 20 percent of dimethyl sulfoxide;

tween-800.01-0.5%;

0.1 to 1.0 percent of preservative;

80.0-99.0% of purified water.

2. A pheromone composition spray formulation according to claim 1, which comprises, by mass:

androstenone and/or androstenol 0.002-0.01%;

herba Epimedii extract 0.002-0.01%;

0.5 to 2.0 percent of dimethyl sulfoxide;

tween-800.02-0.08%;

0.2 to 0.8 percent of preservative;

purified water 95.0-99.0%.

3. A pheromone composition spray formulation according to claim 1, wherein the spray formulation comprises androstenone and androstenol; the mass ratio of androstenone to androstenol is 0.001-0.005% and 0.001-0.005%.

4. A pheromone composition spray formulation according to claim 1, wherein the pH of the spray formulation is in the range of 5 to 8.

5. A pheromone composition spray formulation according to claim 1, which is composed of the following components in mass ratio:

androstenone 0.002%

Androstenol 0.002%;

herba Epimedii extract 0.004%;

1.0% of dimethyl sulfoxide;

tween-800.02%;

0.3 percent of preservative;

98.642% of purified water.

6. A pheromone composition spray formulation according to claim 1, which is composed of the following components in mass ratio:

androstenone 0.002%

Androstenol 0.002%;

herba Epimedii extract 0.002%;

1.0% of dimethyl sulfoxide;

tween-800.05%;

0.3 percent of preservative;

98.644% of purified water.

7. A pheromone composition spray formulation according to claim 1, which is composed of the following components in mass ratio:

androstenol 0.002%;

herba Epimedii extract 0.002%;

1.0% of dimethyl sulfoxide;

tween-800.05%;

0.3 percent of preservative;

98.646% of purified water.

8. A pheromone composition spray formulation according to claim 1, which is composed of the following components in mass ratio:

androstenone 0.002%

Herba Epimedii extract 0.002%;

1.0% of dimethyl sulfoxide;

tween-800.05%;

0.3 percent of preservative;

98.646% of purified water.

9. The process for preparing a pheromone composition spray preparation according to any one of claims 1 to 8, wherein the androstenone, androstenol, and epimedium extract are precisely weighed according to the formula amount, added to dimethyl sulfoxide of the formula amount, and subjected to ultrasonic assisted dissolution for 10 to 30 minutes; adding the mixture into purified water after the mixture is completely dissolved, starting stirring, adding the Tween-80 according to the prescription amount while stirring, continuing stirring, adding the preservative, adding the purified water according to the prescription amount, continuing stirring for 1.5-3 hours, and filtering with a 0.40-0.50 micron filter membrane to obtain the product.

10. Use of a pheromone composition spray formulation according to any one of claims 1 to 8 for stimulating reproductive behavior and increasing reproductive success and productivity in female suid animals.

Technical Field

The invention relates to a pheromone composition, in particular to a pheromone composition spray preparation, a preparation method and application thereof.

Background

The term "Pheromone" was proposed by researchers in the fifties of the last century and was synthesized by the two ancient greek words Pherein and hormone. It is considered to be a chemical species capable of causing an allogenic animal sexual response, and is therefore also known as pheromone, playing an important role in some behavioral and sexual interactions, as well as in the control of reproduction. Pheromones are chemical substances secreted from the exocrine glands of insects to the outside of the body, and act on other individuals of the same species through contact of individual examination or air transmission to cause specific behaviors or physiological responses. The insect sex pheromone can be mainly used for predicting and forecasting pests, directly trapping and killing males, cutting off the relation among pests and the like. The artificially synthesized sex pheromone is called 'sex attractant', and when the sex pheromone is scattered in the field in a large amount, the male insects can get lost and can not mate, thereby achieving the purpose of prevention and control.

At present, there are more than 18 kinds of pheromones and analogues thereof, the chemical structures of which are known and known. However, the pheromones ' androstenone (androst-16-en-3-one) and ' androstenol (androst-16-en-3-ol) ' secreted and released in the body of the boar are found to play an important role in the reproductive behavior of the sow. However, boar androstenone and androstenol have not only been chemically defined, but also have been produced as sprays and powders for use in swine production and for scientific research. According to the past accumulated knowledge, boars sexually stimulate sows before mating. Pheromone, auditory, visual and tactile stimuli are involved in this process, all of which affect the pituitary oxytocin release in sows and replacement gilts (Madej et al, 2005). Langendijk et al (2003) reported that the presence of boars induced oxytocin release in sows and significantly increased the activity of the myometrium in sows. The effects of boar exposure also include stimulation of follicle growth, resulting in oestrus and ovulation in more gilts (Langendijk et al, 2000). The salivary pheromone released by submaxillary gland of 10-month-old boar can stimulate oestrus and oestrus behavior of sow. The pharmaceutical product containing the pig pheromone can be used for enhancing the oestrus expression of sows and detecting the oestrus behavior change of the sows. In practical application, the boar pheromone is found to be used for sows, so that the arrival of the initial period of the sows can be promoted. In addition, studies have shown that boar pheromones can also affect the rhythm and intensity of oestrus in sows and affect the interval between weaning and oestrus. In weaned sows, when they encounter a princess or other form of pheromone stimulation, they develop certain behavioural changes, such as lordosis or orthoreflexes, which signal them to target male proximity, which for the breeder means that they can be bred or inseminated (a.i.).

Estrus refers to the phenomenon of the reproductive cycle of a sexually mature female mammal that manifests itself during a particular season, physiologically as ovulation, ready for fertilization and pregnancy, and behaviorally as attraction and admission of heterosis. Firstly, the ovarian follicle on the ovary is maturing and then ovulatory; female hormones secreted by follicles cause libido and are subject to crawling by males; at the same time, physiological changes such as mucosal congestion, edema, mucus increase, cervix release and the like of the genital catheter occur. The duration of the oestrus behavior and reproductive physiological changes is the oestrus period. The oestrus expression of animals is an important prerequisite for the production of reproductive results. However, in actual production, some hormones, such as estrogen, can be used to induce heat in female animals, but the heat generated in this case is mostly only "false heat", i.e. simple heat, and cannot be bred (artificial insemination) and normal breeding results cannot be generated. When the female animals induced by methods such as estrogen and the like are in estrus, the activity or development state of the ovaries of the female animals is not in a state due to the estrus, or no follicles are developed, or the ovaries do not have normal ovulation function, even corpus luteum exists on the ovaries, and other reproductive organs are not prepared in all aspects of mating and breeding. The action of inducing or promoting the oestrus of the female animals by the pheromone can ensure that the female animals are fertilized, pregnant and farrowing, and the oestrus of the female animals has a normal ovulation function.

Then, factors influencing the breeding of the sows are various, such as oestrus, oestrus identification, semen quality, mating technology, nutritional status, environmental factors and the like, the oestrus identification of the sows is one of important technical links in the breeding work, and the mating period can be correctly determined through the oestrus identification so as to improve the conception rate. The existing estrus identification mostly adopts external observation, and combines methods such as a back pressing test, boar estrus test and the like. However, in actual production, part of sows have normal ovarian function, normal follicular development, normal reproductive system and normal ovulation, but have no oestrus (no oestrus expression or behavior), i.e. recessive oestrus or no obvious oestrus, difficult oestrus identification, mismatch, missed mating and low loss in production. Although external observation is simple, it is difficult to determine the recessive or inconspicuous sow, and the mating period is easily missed. In large-scale production, one sow may not see a few boars from birth to sexual maturity and then mate, and the sow does not contact the boars, which is also a factor for recessive oestrus of the sow. It is also believed that the cause of recessive estrus is due to insufficient estrogen secretion caused by external factors interfering with the normal function of the pituitary.

At present, most pig farms use boars to check whether weaned sows or multiparous sows are in oestrus before sow mating or artificial insemination in order to improve the mating rate; however, even if boars are used for the heat-check method, there is no response in about 20% of sows. In addition, the method has the main problems that a separate colony house is needed for feeding the boars, the boars play a single role in the daily production of a pig farm, and the feeding cost is high. In addition, the disease infection risk is increased by boar inspection, and the boar inspection is a large leak in epidemic prevention links, for example, the epidemic of african pigs spreading China at present, a large part of epidemic situations are caused by that boars randomly pass in and out of a sow colony house, and the sow colony house which is not infected originally is also infected, so that destructive attack is caused to part of pig farms. According to incomplete statistics, about 1/3 sows in China die (fatality of disease + slaughter) due to African swine fever in 2018-2019. The boar oestrus test method is not only complicated, but also not beneficial to the epidemic prevention of the pig farm.

In foreign countries, a bionic method, namely simulating the sound and smell of breeding boars, is adopted to test whether sows are oestrous.

In 1968, UK patent GB1267514A mentions that pheromone has the function of improving artificial insemination of sows, and the product of the pheromone uses diethyl ether as a solvent of androstenone, wherein the diethyl ether has a low boiling point and a flash point of about 34.6 ℃, and is easy to explode when being used and stored at a higher temperature; in addition, the packaging is a pressure bottle, a large amount of alkanes such as propane and butane are filled in the pressure bottle to serve as propellants, the propane and the butane belong to flammable and explosive dangerous chemicals, and particularly, the product has great risks in storage and transportation in hot summer; daily preservation needs to be performed in a shady and cool place or in a refrigerator; in the using process, the paint is volatile and has certain irritant toxicity to respiratory tracts and the like of operators. Therefore, the products are very inconvenient to use and store and have potential safety hazards. The actual effectiveness of the product is low, namely only 55 percent (22/40), and the effect is not ideal, mainly because the product has too fast volatility, the active ingredients are not contacted by the sows, and the active ingredients are volatilized into the ambient air, and the concentration is diluted, so the actual use effect is poor.

Melrose et al, BRITISH VETERINARY JOURNAL 127(1971)497-502, compared the oestrus of sows when different pheromones were used, even after the two steroid hormones 5 α -androst-16-en-3-one and 3 α -hydroxy-5 α -androst-16-ene, the rate of the quiescence reaction (oestrus expression, mating) of sows was only 56% at the highest; C.B. Breeding et al, BRITISH VETERINARY JOURNAL (British VETERINARY science) 130(1974)61-67, studied the use of different androgenic steroid hormones in combination for oestrus detection of sows, when the pheromone is used alone, the proportion of the oestrus expression (mating) of the sows is only 58%.

The bionic method adopted by researchers in China, Heying Jun, oestrus identification of sows [ J ]. Zhejiang animal husbandry and veterinarian, 1995(4) reports that oestrus identification rate is basically the same (276vs 284) with external observation method, but conception rate is obviously higher than that of other methods. Mainly because the sow has the coupling response to the heat identification of the bionic method only when coupling is carried out (namely the full-time estrus), and the sow at the initial stage, the final stage or other sows which do not have heat has no coupling response, therefore, the sow with the coupling response basically has the right mating period, at the moment, the mating has higher conception rate, so the bionic method is not only an effective heat identification method.

A pheromone medicine combination and a method are mentioned in the US9480689B1, which can stimulate the reproductive behavior and the reproductive success rate of sows. The combination comprises at least one steroid hormone and one heterocyclic aromatic compound, after the product is used, the rate of the sow having a standing reaction is 88%, although the clinical effect is obviously improved, the steroid hormone is various, and a plurality of steroid hormones are assimilated and can not be used at will, otherwise the food safety can be influenced, such as estrogen (not beneficial to embryo implantation), androgen (inhibiting follicular development) and the like, if the use time is improper or inaccurate, the reproductive behavior of the sow can not be stimulated, but the reproductive behavior of the female livestock can be inhibited. In addition, heterocyclic aromatic compounds, which are also special, have certain inhalation toxicity besides certain odor, for example, quinoline is an aromatic compound and is combustible by open fire and high heat. Andoxidizing agentThe reaction may take place. Toxic nitric oxide fume is released after heated and decomposed. When the quinoline vapor is used by a user, the quinoline vapor is irritant to the nose and the throat. It can cause headache, dizziness, nausea, and irritation to eyes and skin. In addition, quinoline has certain ecotoxicity, for example, can kill aquatic daphnia, algae and other organisms, and influences the ecological stability of water. The half-life period of biodegradation can reach 960h, and the half-life period of photooxidation in air can reach 99 h. Therefore, if the quinoline is improperly used, the health of users is affected, and certain influence is also caused to the ecological environment. In addition, there have been some researchesThe heterocyclic aromatic compound is found to have a certain growth promoting effect, the growth promoting effect can interfere the growth and development of the female animals and is not beneficial to the development of embryos, the food safety can be influenced, and substances or medicines with the growth promoting effect are strictly prohibited for food animals in many countries. Therefore, the product combination is not perfect enough, and great potential safety hazard also exists.

Therefore, the existing method cannot meet the requirements of the batch production technology of sows in the aspects of effectiveness, safety and the like, namely higher configurable estrus rate.

Disclosure of Invention

In view of the above, it is an object of the present invention to provide a pheromone composition spray preparation which is safer and more reliable, and reliably improves reproductive performance (i.e., higher fertile oestrus rate) of sows, improves utilization rate of sows, and improves fertility of sows.

In order to achieve the above object, the present application adopts the following technical solutions:

a pheromone composition spray preparation, which comprises the following components in percentage by mass:

preferably, the spray formulation comprises, by mass:

the best technical scheme of the composition is as follows: a combination comprising androstenone, androstenol, and epimedium extract. Preferably, the spray formulation comprises androstenone and androstenol; the mass ratio of androstenone to androstenol is 0.001-0.005% and 0.001-0.005%. According to the traditional Chinese medicine, the epimedium herb is pungent, sweet and warm in taste and can walk the liver and kidney meridians. Is an essential herb for nourishing life gate, benefiting vital essence and qi, strengthening bones and muscles, tonifying kidney and strengthening yang, and is clinically used for treating impotence and impotence, spermatorrhea, premature ejaculation, urinary incontinence, infertility of women and other symptoms. The herba Epimedii extract is perennial grassThe main components of the substance extracted from epimedium herb (with the scientific name: Epimediumbrevicornum Maxim.) include icariin,Essential oils、Wax alcohol、Plant sterol、Tanning Quality of food、Vitamin EThe components belong to Chinese herbal medicine extracts, are safer and do not cause adverse stimulation to users and target animals. Icariin (icarin) is preferred, and the purity is more than or equal to 94.0 percent.

Preferably, the formulations herein have a pH in the range of 5 to 8. Between this pH range, an unexpected stabilizing effect of the composition can be obtained. Therefore, the inventor further finds that the pH value is between 5 and 8, the use requirement can be met, when the pH value is lower than 4 or higher than 8, the nasal cavity of the sow can be stimulated to a certain extent, even the nasal mucosa is damaged, the acceptance degree of the sow is poor, in this case, the stability of the medicine is poor, and when the pH value is between 5 and 8, the stability of the solution of the composition is improved and is basically close to the normal condition of the body, and when the composition is used, the acceptance degree of the sow is high.

As a specific embodiment, the spray preparation is composed of the following components in mass ratio:

as a specific embodiment, the spray preparation is composed of the following components in mass ratio:

as a specific embodiment, the spray preparation is composed of the following components in mass ratio:

as a specific embodiment, the spray preparation is composed of the following components in mass ratio:

the application also discloses a preparation method of the pheromone composition spray preparation, which comprises the steps of precisely weighing androstenone, androstenol and epimedium herb extracts according to the formula amount, adding the androstenone, androstenol and epimedium herb extracts into dimethyl sulfoxide of the formula amount, and performing ultrasonic assisted dissolution for 10-30 minutes; adding the mixture into purified water after the mixture is completely dissolved, starting stirring, adding the Tween-80 according to the prescription amount while stirring, continuing stirring, adding the preservative, adding the purified water according to the prescription amount, continuing stirring for 1.5-3 hours, and filtering with a 0.40-0.50 micron filter membrane to obtain the product.

The application also discloses the use of said pheromone composition spray formulation for stimulating reproductive behavior and increasing reproductive success and productivity in female suid animals. The technical scheme of the application reliably improves the reproductive performance (namely higher matchable estrus rate) of the sow, improves the utilization rate of the sow and improves the fertility of the sow.

Drawings

FIG. 1 is a different type of spray apparatus; a and B are two different mechanical spray bottles; c, D is an electric spraying device.

FIG. 2 is a graph of the actual effectiveness increase for different pheromone processing groups; remarking: androstenone, 5 α; androstenol, 3 α; androstenone + androstenol, 5 α +3 α; epimedium 1, Y-1; epimedium 2, Y-2; androstenone + epimedium 1, 5 alpha + Y-1; androstenol + epimedium 1, 3 alpha + Y-1; androstenone + androstenol + epimedium 1, 5 alpha +3 alpha + Y-1; androstenone + androstenol + epimedium 2, 5 alpha +3 alpha + Y-2.

FIG. 3 is a schematic diagram of distribution of different piggeries in the same house in the same pig farm (I-VI represents distribution of different piggeries).

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is illustrated by the following specific embodiments and examples, which are intended to better explain the present invention without limiting the scope of the present invention, and all modifications and variations that are within the basic idea and principle of the present invention are included in the scope of the present invention as claimed.

Preparation examples

The following terms describe the composition of androstenone, androstenol and epimedium extract in specific mixing ratios in the examples of the invention.

Comparative example 1

20.0mg of androstenone (Sigma-Aldrich,. gtoreq.98.00%) was added to 10.0g of dimethyl sulfoxide (Sigma-Aldrich, chromatographic grade) according to the formulation amount of the pheromone-containing composition solution listed in Table 1, and the mixture was subjected to ultrasonic assisted dissolution for 20 minutes; after complete dissolution, the solution was added to 500ml of MilliQ-water, stirring was started, 0.5g of Tween-80 (Sigma-Aldrich) was added while stirring, then stirring was continued, 3g of preservative (Caron, LONZA, Switzerland) was added, then MilliQ-water was added to 1000.0g, stirring was continued for 2 hours, and finally filtration was performed using a 0.45 micron filter (Merk-Millipore) to obtain composition A solution. Before use, the solution was dispensed into a spray device at 100ml per bottle.

TABLE 1 pheromone composition solution formulations

Components	Percent by weight	g
			Androstenone	0.002	0.002
Dimethyl sulfoxide	1.0	1.0
			Tween-80	0.05	0.05
Preservative	0.3	0.3
			Purified water	98.648	98.648
Total of	100.000	100.000

Comparative example 2

According to the formulation of the composition containing pheromone androstenol (Sigma-Aldrich,. gtoreq.98.00%) as listed in Table 2. Adding 20.0mg of androstenol into 10.0g of dimethyl sulfoxide, and performing ultrasonic assisted dissolution for 20 minutes; after the dissolution, the mixture was added to 500ml of MilliQ-water, stirring was started, 0.5g of Tween-80 was added while stirring, then stirring was continued, 3g of preservative (Kathon, LONZA, Switzerland) was added, MilliQ-water was added to 1000.0g, stirring was continued for 2 hours, and finally filtration was performed using a 0.45 micron filter (Merk-Millipore) to obtain a solution of composition B. Before use, the solution was dispensed into a spray device at 100ml per bottle.

TABLE 2 pheromone composition solution formulations

Components	Percent by weight	g
			Androstenol	0.002	0.002
Dimethyl sulfoxide	1.0	1.0
			Tween-80	0.05	0.05
Preservative	0.3	0.3
			Purified water	98.648	98.648
Total of	100.000	100.000

Comparative example 3

The formulation of the solution containing pheromone androstenone and androstenol composition is shown in table 3. 1000ml of the C composition solution was prepared by following the solution preparation methods of examples 1 and 2. Before use, the solution was dispensed into a spray device at 100ml per bottle.

TABLE 3 pheromone composition solution formulations