A pheromone composition for improving the estrus efficiency of cows and a method for preparing the same

Abstract

The present application relates to a kind of pheromone composition for improving cow estrus efficiency and its preparation method, the pheromone composition includes 0.0003~3wt% bull pheromone, 0.0001~10wt% phenolic compound and 87~99.9996wt% carrier solvent;The bull pheromone is one or more of acetic acid, trimethylamine, 1-iodo undecane, and the phenolic compound is tetramethylphenol.The preparation method of the pheromone composition is as follows: bull pheromone, phenolic compound is added into carrier solvent, material is dissolved after stirring uniformly, and semi-finished product solution is obtained;Nitrogen is filled in filling bottle, and then semi-finished product solution is filled into bottle, and is sealed, to obtain pheromone composition spray preparation finished product.The smell of the pheromone composition is closer to the smell emitted from bull, can better stimulate cow, excite sexual behavior, and improve estrus rate.

Description

Technical Field

[0001] This invention relates to the field of animal husbandry technology, and in particular to a pheromone composition for improving the estrus efficiency of cows and its preparation method. Background Technology

[0002] Accurate identification of estrus in cows is a prerequisite for efficient breeding on cattle farms. Timely artificial insemination is only successful during the estrus period, mainly because accurate estrus detection can prevent cows from being in heat but not being bred, thus reducing the period of non-pregnancy in cows.

[0003] In production, the quiet estrus behavior of cattle is not obvious, making identification difficult and impacting the economic benefits of cattle farms. During mating between bulls and cows, sensory stimulation such as touch, sight, hearing, and smell stimulates the pituitary gland of the cow, inducing estrus and ovulation. Firstly, farms typically have few bulls, requiring significant manpower and resources. Using bull pheromone sprays for estrus detection can reduce production costs and ensure biosecurity.

[0004] Early reports on pedometer-based estrus detection methods using accelerometers have largely achieved automation. However, numerous studies have shown that over 20% of cows in estrus show no significant increase in activity, indicating that latent estrus necessitates further improvement and refinement of pedometer-based estrus detection. Research has identified several estrus-specific pheromones in cow feces, urine, vaginal fluids, saliva, and milk secretions. Archunan found acetic acid, propionic acid, and 1-iodoundecane in the feces of Jersey cows during estrus, suggesting they may be contributing factors to olfactory reflexes and mounting behaviors in bulls. Sankar et al. detected 13 pheromones in the saliva of cows in estrus and, after validation with bulls, found that trimethylamine, acetic acid, propionic acid, valeric acid, and tetrapropylphenol elicited increased olfactory reflexes and mounting frequency in bulls. The response to trimethylamine was significantly higher than to other compounds, warranting further investigation.

[0005] Currently, there are few patent reports on boar pheromone composition sprays. Samuthirapandi et al. developed a buffalo tetramethylphenol and 9-octadecenoic acid estrus detection kit in 2017, but it has not been widely used in large-scale cattle farms. There are more reports on porcine pheromone composition sprays. For example, Chinese patents CN108289898B and CN111818926A both mention a three-molecule pheromone composition composed of androstenone, androstenol, and quinoline for estrus detection in sows; the corresponding product has been commercialized under the brand name Fairmont (Villon Asco, Italy). Chinese patent application CN111467309A mentions a spray for boar estrus detection containing estradiol. Since the impact of African swine fever in 2018, boar pheromone compositions have gradually been promoted and used in large-scale pig farms. Summary of the Invention

[0006] Technical problem solved: In view of the shortcomings of the existing technology, the present invention provides a pheromone composition for improving the estrus efficiency of cows and its preparation method. The pheromone composition has an odor that is closer to the smell emitted by bulls, which can better stimulate cows, arouse sexual behavior, and increase the estrus rate.

[0007] Technical solution: A pheromone composition for improving the estrus efficiency of cows, the pheromone composition comprising 0.0003-3 wt% bull pheromone, 0.0001-10 wt% phenolic compound and 87-99.9996 wt% carrier solvent; wherein the bull pheromone is one or more of acetic acid, trimethylamine, and 1-iodoundecane, and the phenolic compound is tetramethylphenol.

[0008] The aforementioned carrier solvent is composed of pure water and alcohol solvent, wherein the alcohol solvent accounts for 4.8% to 5.1% of the volume percentage of the carrier solvent.

[0009] The alcohol solvents mentioned above are one or more of ethanol, ethylene glycol, propanol, isopropanol, butanol, pentanol, hexanol, heptanol, and octanol.

[0010] The bull pheromone mentioned above is composed of acetic acid, trimethylamine, and 1-iodoundecane.

[0011] The pheromone composition described above comprises 0.0001–1 wt% acetic acid, 0.0001–1 wt% trimethylamine, 0.0001–1 wt% 1-iodoundecane, 0.0001–10 wt% phenolic compounds, and 87–99.9996 wt% transport solvent.

[0012] The pheromone composition described above comprises 0.0005–0.2 wt% acetic acid, 0.0005–0.2 wt% trimethylamine, 0.0005–0.2 wt% 1-iodoundecane, 0.0002–5 wt% tetramethylphenol, and 94.4–99.9983 wt% carrier solvent.

[0013] The pheromone composition described above comprises 0.001–0.1 wt% acetic acid, 0.001–0.1 wt% trimethylamine, 0.001–0.1 wt% 1-iodoundecane, 0.0004–0.5 wt% tetramethylphenol, and 99.2–99.9966 wt% carrier solvent.

[0014] The pheromone composition described above is formulated as a spray.

[0015] The method for preparing the pheromone composition for improving estrus efficiency in cows described above is characterized by the following steps:

[0016] Step 1: Add bull pheromones and phenolic compounds to the carrier solvent. After the materials melt, stir evenly to obtain a semi-finished solution.

[0017] Step 2: Fill the filling bottle with nitrogen, then fill the bottle with the semi-finished solution, seal the bottle, and obtain the finished pheromone composition spray formulation.

[0018] Beneficial effects: The pheromone composition and its preparation method for improving estrus efficiency in cows provided by this invention have the following beneficial effects:

[0019] 1. The pheromone composition of the present invention contains tetramethylphenol, which can effectively induce cows to actively inhale the pheromone composition, improve estrus efficiency, and enhance the reproductive and fertility of cows;

[0020] 2. The bull pheromone of the present invention is one or more of acetic acid, trimethylamine, and 1-iodoundecane, which makes the smell of the composition closer to the smell emitted by bulls, which can better stimulate cows, arouse sexual behavior, and increase estrus rate;

[0021] 3. The pheromone composition of the present invention is in the form of a spray formulation, which is simple to operate when applied to cows. Attached Figure Description

[0022] Figure 1 This is a graph showing the effect of tetramethylphenol on the behavior of cows actively sniffing pheromones in Experiment 1.

[0023] Figure 2 This is a graph showing the effect of tetramethylphenol addition on the behavior of cows actively seeking out pheromones in Experiment 2.

[0024] Figure 3 This is a graph showing the effects of different pheromone combinations on the behavior of cow Flame in Experiment 3.

[0025] Figure 4 This is a graph showing the effects of different pheromone combinations on the fertility and reproductive efficiency of cows in Experiment 4. Detailed Implementation

[0026] The technical solution of the present invention will be further described and illustrated below through specific embodiments. It should be understood that the specific embodiments described herein are only for the purpose of helping to understand the present invention and are not intended to limit the present invention. Unless otherwise specified, the raw materials used in the embodiments of the present invention are all commonly used raw materials in the art, and the methods used in the embodiments are all conventional methods in the art.

[0027] The materials used in the following examples were sourced as follows: acetic acid (≥98.5%), trimethylamine (98.00%), 1-iodoundecane (≥98%), and tetramethylphenol (≥98%) were all purchased from Sangon-Biotech; ethanol (97%, AR grade) and isopropanol (≥99%) were purchased from Tacklan Technology Co., Ltd.; and purified water was produced by Sichuan Youpu Technology Co., Ltd. using an ultrapure water system.

[0028] The components and content of each component of the pheromone compositions of Examples 1-10 and Comparative Examples 1-3 are shown in Table 1 below.

[0029] Table 1

[0030]

[0031] Note: The content of each component is in wt%.

[0032] Example 1

[0033] Weigh each component according to the proportions in Example 1 in Table 1. Add the alcohol solvent (ethanol) to the mixing bottle, and then add bull pheromones (acetic acid, trimethylamine, 1-iodoundecane) and tetramethylphenol in sequence according to the proportions. Stir at 400 rpm for 30 minutes to completely dissolve the materials. Then add purified water and continue stirring at 400 rpm for 30 minutes to obtain a uniformly mixed semi-finished solution for later use. Next, purge the filling bottle with nitrogen to achieve an oxygen-free condition, and then fill the prepared semi-finished solution into the bottle, cap and seal it to obtain the finished pheromone composition spray.

[0034] Examples 2-4

[0035] The components and contents of Examples 2-4 are shown in Table 1, and the preparation methods are the same as those of Example 1.

[0036] Examples 5-6

[0037] The components and contents of Examples 5-6 are shown in Table 1, and the preparation methods are the same as those of Example 1.

[0038] The alcohol solvents used in Examples 5-6 were ethanol and isopropanol.

[0039] Example 7

[0040] The components and contents of Example 7 are shown in Table 1, and the preparation method is the same as that of Example 1.

[0041] The alcohol solvent used in Example 7 was isopropanol.

[0042] Example 8

[0043] The components and contents of Example 8 are shown in Table 1, and the preparation method is the same as that of Example 1.

[0044] The bull pheromone used in Example 8 was composed of trimethylamine and 1-iodoundecane.

[0045] Example 9

[0046] The components and contents of Example 9 are shown in Table 1, and the preparation method is the same as that of Example 1.

[0047] The bull pheromone used in Example 9 was composed of acetic acid and 1-iodoundecane.

[0048] Example 10

[0049] The components and contents of Example 10 are shown in Table 1, and the preparation method is the same as that of Example 1.

[0050] The bull pheromone used in Example 10 is composed of acetic acid and trimethylamine.

[0051] Comparative Example 1

[0052] The components and contents of Comparative Example 1 are shown in Table 1, and the preparation method is the same as that of Example 1.

[0053] Comparative Examples 2-3

[0054] The components and contents of Comparative Examples 2-3 are shown in Table 1, and the preparation methods are the same as those in Example 1.

[0055] Comparative Examples 2-3 did not contain bull pheromones (acetic acid, trimethylamine, 1-iodoundecane).

[0056] The performance of the pheromone composition for improving estrus efficiency in cows according to the above embodiments and comparative examples was tested. The test process and results are as follows.

[0057] Experiment 1 determined the effect of tetramethylphenol on inducing cows to actively approach and sniff pheromones.

[0058] 1.1 Experimental Objective

[0059] The purpose of this experiment is to determine whether tetramethylphenol can effectively induce cows to actively seek out pheromones, and to determine whether tetramethylphenol is the only effective ingredient in the pheromone composition that induces cows to actively seek out pheromones.

[0060] 1.2 Test Plan

[0061] This experiment was conducted on a large-scale cattle farm with 195 healthy postpartum cows. All cows were kept in the same exercise area, with balanced nutrition and free access to water. A 2.8m high frame was set up in the center of the exercise area, with a 1×1m area placed horizontally on top. 2A carrier steel plate with a thickness of 0.2 cm was used. Different compositions of the same components (the compositions of Example 4 and Comparative Examples 1-2) and pure water were sprayed onto the center point of the carrier steel plate from a distance of about 10 cm perpendicular to the carrier steel plate. The number of cows that moved toward the carrier steel plate within 30 seconds was then observed, and the number of cows that exhibited sniffing behavior on the surface of the carrier steel plate (the nose and lips mirror contacted the center point of the carrier steel plate) was also recorded.

[0062] 1.3 Test Results

[0063] The effects of tetramethylphenol on the behavior of cows are shown in Table 2 below. Figure 1 As shown. Data significance was analyzed using SPSS 26 software; percentage data were analyzed using the χ² method. 2 The significance of the test was statistically analyzed, and P < 0.05 indicated a significant difference.

[0064] Table 2

[0065] Group number Number of test heads Number of moving heads Number of sniff probes Mobility rate (%) Sniffing rate (%) D 56 43 27 <![CDATA[76.79 a ]]> <![CDATA[48.21 a ]]> K 44 15 9 <![CDATA[34.09 b ]]> <![CDATA[20.45 b ]]> L 48 33 21 <![CDATA[68.75 a ]]> <![CDATA[43.75 a ]]> purified water 47 6 3 <![CDATA[12.77 b ]]> <![CDATA[6.38 b ]]>

[0066] Note: Different subscript letters for a and b indicate significant differences in data (P<0.05), while the same subscript letter indicates no significant differences in data (P>0.05). Wherein: Motion rate = (Number of moving heads / Number of test heads) × 100%; Smell detection rate = (Number of smegapixel probes / Number of test heads) × 100%.

[0067] From Table 2 and Figure 1 It can be seen that the movement rate and sniffing rate of groups D and L were significantly higher than those of group K and pure water (P<0.05), while there was no significant difference between groups D and L (P>0.05). This indicates that tetramethylphenol in the composition can effectively induce cows to sniff the carrier steel plate for pheromones, and tetramethylphenol is the only component in the pheromone composition of this invention with this function.

[0068] Screening of tetramethylphenol concentration in Experiment 2

[0069] 2.1 Experimental Objective

[0070] The purpose of this experiment was to determine the optimal amount of tetramethylphenol added when using the pheromone composition to induce cows to actively sniff pheromones.

[0071] 2.2 Test Plan

[0072] This experiment involved 156 healthy postpartum cows. All cows were kept in the same exercise area, received balanced nutrition, and had free access to water. A 2.8m high frame was set up in the center of the exercise area, with a 1×1m area placed horizontally on top. 2A carrier steel plate with a thickness of 0.2 cm was used. Different compositions of the same composition (the composition sprays in Examples 1-4) were sprayed onto the center point of the carrier steel plate from a distance of about 10 cm perpendicular to the carrier steel plate. The number of cows that moved toward the carrier steel plate within 30 seconds was then observed, and the number of cows that exhibited sniffing behavior on the surface of the carrier steel plate (the nose and lips mirror contacted the center point of the carrier steel plate) was also recorded.

[0073] 2.3 Test Results

[0074] The effects of tetramethylphenol on the behavior of cows are shown in Table 3 below. Figure 2 As shown. Data significance was analyzed using SPSS 26 software; percentage data were analyzed using the χ² method. 2 The significance of the test was statistically analyzed, and P < 0.05 indicated a significant difference.

[0075] Table 3

[0076] Group number Number of test heads Number of moving heads Number of sniff probes Mobility rate (%) Sniffing rate (%) A 40 11 7 <![CDATA[27.50 c ]]> <![CDATA[17.50 c ]]> B 35 23 16 <![CDATA[65.71 b ]]> <![CDATA[45.71 b ]]> C 37 27 24 <![CDATA[72.97 ab ]]> <![CDATA[64.86 a ]]> D 44 34 17 <![CDATA[77.27 a ]]> <![CDATA[38.64 b ]]>

[0077] Note: Different subscript letters for a and b indicate significant differences in data (P<0.05), while the same subscript letter indicates no significant differences in data (P>0.05). Wherein: Motion rate = (Number of moving heads / Number of test heads) × 100%; Smell detection rate = (Number of smegapixel probes / Number of test heads) × 100%.

[0078] From Table 3 and Figure 2 It can be seen that the movement rates of groups C and D were 72.97% and 77.27%, respectively, with no significant difference (P>0.05). However, the movement rates of group D were significantly different from those of groups A and B (P<0.05). This indicates that a tetramethylphenol addition of 0.4% has the best effect in inducing cows to move towards the carrier plate. The sniffing rates of groups D and C were 38.64% and 64.86%, respectively, with a significant difference (P<0.05), indicating that a addition of 0.04% has the best effect in inducing cows to sniff the carrier plate. Furthermore, the sniffing rates of group C were also significantly different from those of groups A and B (P<0.05), with the sniffing rate of group B being significantly higher than that of group A (P<0.05). This indicates that when the addition is reduced to 0.0004%, the effect of inducing cows to move towards and sniff the carrier plate is significantly reduced.

[0079] Experiment 3: Effects of pheromone combinations on the behavior of cow Flame

[0080] 3.1 Experimental Objective

[0081] The purpose of this experiment was to determine whether different bull pheromone compositions could enhance Flam behavior in cows during estrus.

[0082] 3.2 Test Plan

[0083] All breeding cows in this experiment were kept in the same exercise area, with balanced nutrition and free access to water. Different compositions (compositions from Examples 3, 8-10, and Comparative Example 3) were sprayed onto the faces of the cows. During or after the spraying, cows exhibiting Flame behavior were sprayed a second time. The frequency and timing of sniffing, licking of nostrils, urination, and mounting were observed and recorded within 5-10 minutes.

[0084] 3.3 Test Results

[0085] The effects of different bull pheromone compositions on the behavior of cow Flame, such as Figure 3 As shown. Data are expressed as mean ± standard deviation. Significance was determined using SPSS 26 software. P < 0.05 indicates a significant difference, and P < 0.01 indicates an extremely significant difference.

[0086] from Figure 3 It can be seen that the Flame behavior in group C was significantly higher than that in other groups (P<0.01), and that group HJ was significantly higher than that in group M and purified water (P<0.05), while there was no significant difference between group M and purified water (P>0.05). Therefore, the combined effect of acetic acid, trimethylamine, and 1-iodoundecane is significantly better than the use of bull pheromones alone, effectively inducing Flame behavior in cows and improving estrus efficiency.

[0087] Experiment 4: Effects of different pheromone combinations on fertility and reproductive efficiency in cows

[0088] 4.1 Experimental Objective

[0089] The purpose of this experiment was to determine whether compositions containing different bull pheromones could enhance fertility and reproductive efficiency in cows during estrus detection.

[0090] 4.2 Test Plan

[0091] This experiment involved 320 healthy postpartum cows, randomly grouped and provided with balanced nutrition and free access to water. Day 1 was recorded as the start of estrus detection. Different compositions (compositions from Examples 3, 6-10, and Comparative Example 3) were sprayed onto the cows' faces twice daily, once in the morning and once in the afternoon. Estrus detection was performed promptly. If a cow exhibited mounting behavior and was receptive to mounting, a first uterine insemination was performed 8-10 hours later, followed by a second insemination 8-10 hours later. If estrus characteristics persisted, spraying continued. Estrus detection was completed within 15 days of the estrus cycle. The number of cows exhibiting estrus was accurately recorded, and the number of pregnant cows was checked by ultrasound 40-45 days after the second insemination.

[0092] 4.3 Test Results

[0093] The effects of different bull pheromone compositions on the fertility and reproductive efficiency of cows are shown in Table 4 below. Figure 4 As shown. Data significance was analyzed using SPSS 26 software; percentage data were analyzed using the χ² method. 2 The significance of the test was statistically analyzed, and P < 0.05 indicated a significant difference.

[0094] Table 4

[0095]

[0096]

[0097] Note: Different subscript letters in a and b indicate significant differences (P<0.05), while the same subscript letter indicates no significant difference (P>0.05). Wherein: Estrus rate = (Number of heads in estrus / Number of heads tested) × 100%; Conception rate = (Number of heads conceived / Number of heads in estrus) × 100%.

[0098] From Table 4 and Figure 4 It can be seen that the estrus rates of groups G and M were significantly lower than those of other experimental groups. This indicates that the pheromone composition containing tetramethylphenol can induce cows to actively inhale pheromones, thereby increasing their estrus and conception rates. Furthermore, using the pheromone composition of this invention can improve the reproductive behavior and fertility of cows during estrus detection while reducing the workload of farm workers.

[0099] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

Claims

1. A pheromone composition for improving the efficiency of estrus in cows, characterized in that: The pheromone composition comprises 0.0001–1 wt% acetic acid, 0.0001–1 wt% trimethylamine, 0.0001–1 wt% 1-iodoundecane, 0.0001–10 wt% tetramethylphenol, and 87–99.9996 wt% carrier solvent; the carrier solvent is composed of purified water and an alcohol solvent, wherein the alcohol solvent accounts for 4.8%–5.1% of the volume percentage of the carrier solvent, and the alcohol solvent is one or more selected from ethanol, ethylene glycol, propanol, isopropanol, butanol, pentanol, hexanol, heptanol, and octanol.

2. The pheromone composition for improving estrus efficiency in cows according to claim 1, characterized in that: The pheromone composition comprises 0.0005–0.2 wt% acetic acid, 0.0005–0.2 wt% trimethylamine, 0.0005–0.2 wt% 1-iodoundecane, 0.0002–5 wt% tetramethylphenol, and 94.4–99.9983 wt% carrier solvent.

3. The pheromone composition for improving estrus efficiency in cows according to claim 2, characterized in that: The pheromone composition comprises 0.001–0.1 wt% acetic acid, 0.001–0.1 wt% trimethylamine, 0.001–0.1 wt% 1-iodoundecane, 0.0004–0.5 wt% tetramethylphenol, and 99.2–99.9966 wt% carrier solvent.

4. The pheromone composition for improving estrus efficiency in cows according to claim 1, characterized in that: The pheromone composition is formulated as a spray.

5. A method for preparing a pheromone composition for improving estrus efficiency in cows as described in any one of claims 1-4, characterized in that... The steps are as follows: Step 1: Add bull pheromones and phenolic compounds to the carrier solvent. After the materials melt, stir evenly to obtain a semi-finished solution. Step 2: Fill the filling bottle with nitrogen gas, then fill the bottle with the semi-finished solution, seal the bottle, and obtain the finished product of the pheromone composition spray formulation.

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