Odinagogue for large-scale breeding of bufo bufo gargarizans and spawning induction method thereof
By using Ringer's solution, domperidone and progesterone-releasing hormone A2 oxytobacterium, combined with specific incubation conditions, the low egg laying rate and inconcentration of time in artificial large-scale breeding of Chinese toads were solved, and efficient induced and hatching effects were achieved, and large-scale artificial breeding of Chinese toads was promoted.
Patent Information
- Application Number
- CN202510483891.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-17
- Publication Date
- 2025-07-08
AI Technical Summary
In artificial large-scale breeding of Chinese toads, there are problems such as large amount of toads, low natural egg laying rate, inconcentrated egg laying time, and out-synchronized seedling development, which affects the large-scale artificial reproduction efficiency of medicinal Chinese toads.
Ringer's solution, domperidone and progesterone-releasing hormone A2 are used as oxytoxins, and the abdomen of the Chinese toads are injected subcutaneously into the abdomen of the Chinese toads, combined with male and female toads to hold and lay eggs, and then the egg belt is incubated at a specific temperature to achieve induced childbirth and hatching.
The yield rate reaches 95-98%, the concentrated egg laying rate is high, the seedling survival rate is high, the specifications are neat, and it is easy to manage, which has promoted the large-scale artificial breeding of the Chinese toad and the development of the pharmaceutical industry.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of agricultural breeding, in particular to the technical field of Chinese giant toad breeding, and relates to an oxytocin for large-scale breeding of Chinese giant toad and an oxytocin method thereof. Background Art
[0002] Toad venom is a traditional and precious animal medicinal material in my country. The Chinese giant toad Bufo bufo gargarizans Cantor is one of the original animals of toad venom. According to the "Notice of the State Forestry and Grassland Administration on Standardizing the Classification and Management Scope of Prohibited Wild Animals" (Linhufa
[2020] No. 90), the breeding of Chinese toad Bufo gargarizans for the purpose of eating is prohibited, but breeding for non-food purposes such as medicine, display, and scientific research is allowed.
[0003] Against the background that the country allows the artificial breeding of Chinese giant toads for medicinal purposes, large-scale breeding of Chinese giant toads has gradually emerged in Shandong, Anhui and other places. However, in the process of artificial large-scale breeding of Chinese giant toads, there are common problems such as large amount of breeding toads, low natural egg-laying rate, unconcentrated egg-laying time, and asynchronous development of seedlings. Summary of the invention
[0004] In view of the above-mentioned shortcomings of the prior art, the purpose of the present invention is to provide an oxytocin and an oxytocin method for large-scale breeding of Chinese toads, so as to improve the efficiency of artificial breeding of Chinese toads.
[0005] To achieve the above-mentioned and other related purposes, the present invention provides, in a first aspect, an oxytocin for large-scale breeding of Bufo bufo gargarizans, comprising Ringer's solution, domperidone (DOM), and luteinizing hormone-releasing hormone A2 (LRH-A2).
[0006] The second aspect of the present invention provides a method for preparing an oxytocin for large-scale breeding of Chinese giant toads, comprising: mixing the components of the oxytocin for large-scale breeding of Chinese giant toads provided by the first aspect of the present invention in a ratio to provide the required oxytocin.
[0007] A third aspect of the present invention provides a use of an oxytocin for large-scale breeding of Bufo bufo gargarizans in artificial induction of labor in Bufo bufo gargarizans.
[0008] The fourth aspect of the present invention provides a method for inducing labor in large-scale breeding of Chinese giant toads, comprising: subcutaneously injecting the oxytocin for large-scale breeding of Chinese giant toads provided by the first aspect of the present invention into the abdomen of Chinese giant toads, taking female Chinese giant toads and male Chinese giant toads and placing them in a breeding box for pairing and laying eggs.
[0009] The fifth aspect of the present invention provides an incubation method for the large-scale reproduction of Bufo gargarizans, which includes: performing spawning by using the oxytocic method for the large-scale reproduction of Bufo gargarizans provided in the fourth aspect of the present invention, collecting the egg ribbons and putting them into a breeding pond for incubation to hatch tadpole seedlings.
[0010] As described above, the oxytocic agent and the oxytocic method for the large-scale reproduction of Bufo gargarizans provided by the present invention have the following beneficial effects:
[0011] (1) The oxytocic agent and the oxytocic method for the large-scale reproduction of Bufo gargarizans provided by the present invention adopt the oxytocic agent and the artificial oxytocic method, and the oxytocic rate for Bufo gargarizans is 95-98%. It can solve the problems of large consumption of breeding toads, low natural spawning rate, non-concentrated spawning time, and asynchronous development of fry during the process of artificial large-scale breeding of Bufo gargarizans.
[0012] (2) The oxytocic agent and the oxytocic method for the large-scale reproduction of Bufo gargarizans provided by the present invention improve the reproduction efficiency of Bufo gargarizans, can realize the large-scale artificial reproduction of Bufo gargarizans, effectively promote the development of the artificial large-scale breeding industry of medicinal Bufo gargarizans, and have important popularization significance.
[0013] (3) The oxytocic agent and the oxytocic method for the large-scale reproduction of Bufo gargarizans provided by the present invention have the advantages that the artificially reproduced tadpole seedlings of Bufo gargarizans have high survival rate, uniform specifications, convenient management, and long growth cycle during the tadpole stage, and have important significance in aspects such as the development of amphibian reproduction technology and genetic breeding for the artificial large-scale breeding of medicinal Bufo gargarizans. Specific Embodiments
[0014] The first aspect of the present invention provides an oxytocic agent for the large-scale reproduction of Bufo gargarizans, which includes Ringer's solution, domperidone (DOM), and luteinizing hormone-releasing hormone A2 (LRH-A2).
[0015] In an embodiment, in the oxytocic agent for the large-scale reproduction of Bufo gargarizans, the ratio of the volume (mL) of Ringer's solution to the mass (mg) of domperidone (DOM) is 1.0:1-10, specifically such as 1.0:1-2, 1.0:2-3, 1.0:3-10, and the most preferred is 1.0:5.
[0016] In an embodiment, in the oxytocic agent for the large-scale reproduction of Bufo gargarizans, the ratio of the volume (mL) of Ringer's solution to the mass (μg) of luteinizing hormone-releasing hormone A2 (LRH-A2) is 1.0:1-10, specifically such as 1.0:1-2, 1.0:2-3, 1.0:3-10, and the most preferred is 1.0:5.
[0017] In one embodiment, the Ringer's solution is the commonly used compound sodium chloride injection. Specifically, the Ringer's solution is obtained by adding potassium chloride and calcium chloride to physiological saline.
[0018] In one embodiment, the CAS number of domperidone (DOM) is 57808-66-9.
[0019] In one embodiment, the CAS number of luteinizing hormone-releasing hormone A2 (LRH-A2) is 9034-40-6.
[0020] The second aspect of the present invention provides a method for preparing an oxytocic agent for large-scale reproduction of Bufo gargarizans, including: taking the components in the ratio of the oxytocic agent for large-scale reproduction of Bufo gargarizans provided in the first aspect of the present invention and mixing them to provide the required oxytocic agent.
[0021] The third aspect of the present invention provides the use of an oxytocic agent for large-scale reproduction of Bufo gargarizans in artificial oxytocin induction of Bufo gargarizans.
[0022] The fourth aspect of the present invention provides an oxytocin induction method for large-scale reproduction of Bufo gargarizans, including: subcutaneously injecting the oxytocic agent for large-scale reproduction of Bufo gargarizans provided in the first aspect of the present invention into the abdominal part of Bufo gargarizans, and placing female Bufo gargarizans and male Bufo gargarizans together in a breeding box for embracing and spawning.
[0023] In one embodiment, the Bufo gargarizans selected are well-developed female and male breeding toads.
[0024] In a preferred embodiment, the development requirements for the female Bufo gargarizans are: well-developed gonads, enlarged abdomen, and a granular feeling when touched by hand.
[0025] In a preferred embodiment, the development requirements for the male Bufo gargarizans are: large size, well-developed gonads, healthy physique, and good vitality.
[0026] In one embodiment, during the subcutaneous injection, use forceps to pick up the abdominal skin of Bufo gargarizans and insert the needle at an angle of 40-50 degrees, preferably 45 degrees.
[0027] In one embodiment, during the subcutaneous injection, do not insert the syringe needle into the abdominal cavity of Bufo gargarizans.
[0028] In one embodiment, when the Bufo gargarizans is female, the injection volume for the female Bufo gargarizans is 0.4-0.6 mL / individual, specifically such as 0.4-0.5 mL / individual, 0.5-0.6 mL / individual, and preferably 0.5 mL / individual.
[0029] In one embodiment, when the Chinese giant toad is male, the injection volume for the male Chinese giant toad is 0.2 - 0.3 mL per toad, specifically such as 0.2 - 0.25 mL per toad, 0.25 - 0.3 mL per toad, and preferably 0.25 mL per toad.
[0030] In one embodiment, the quantity ratio of female Chinese giant toads to male Chinese giant toads in the same breeding box is 1:1 - 2, and preferably 1:1 - 1.5.
[0031] In one embodiment, the breeding box is a hollow box with an open top for conventionally breeding Chinese giant toads. The material of the breeding box is plastic.
[0032] In one embodiment, water is added to the breeding box, and the water depth in the breeding box is 10 - 30 cm, preferably 10 - 20 cm.
[0033] In one embodiment, fresh branches are placed in the breeding box. Placing fresh branches is beneficial for female toads to lay eggs.
[0034] In a preferred embodiment, the branches are selected from at least one of willow branches or peach branches.
[0035] In a preferred embodiment, the quantity of branches placed is 5 - 8 for every 40 pairs of female and male Chinese giant toads.
[0036] In one embodiment, the egg - laying time is not less than 24 h after female Chinese giant toads and male Chinese giant toads are placed together in the breeding box, preferably 24 - 48 h.
[0037] The fifth aspect of the present invention provides an incubation method for large - scale breeding of Chinese giant toads, including: performing egg - laying by using the oxytocic method for large - scale breeding of Chinese giant toads provided in the fourth aspect of the present invention, collecting the egg ribbons and putting them into a breeding pond for incubation to hatch tadpole seedlings.
[0038] In one embodiment, the water temperature in the breeding pond is 12 - 18 °C.
[0039] In one embodiment, the incubation time is 10 - 20 days, preferably 14 - 18 days, and more preferably 14 days.
[0040] The water temperature in the above - mentioned breeding pond is negatively correlated with the incubation time. When the water temperature in the breeding pond is higher, the incubation time is shorter, and vice versa.
[0041] For the technical solutions provided by the present invention, unless otherwise specified, the reagents or materials used can be obtained commercially.
[0042] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the protection scope of the present invention.
[0043] The following specific examples illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.
[0044] Example 1
[0045] Take each component and mix them. Each component includes 30 mL of Ringer's solution, 150 mg of domperidone, and 0.15 mg of luteinizing hormone-releasing hormone A2, to obtain the oxytocic agent sample 1# for the large-scale reproduction of Bufo gargarizans.
[0046] Example 2
[0047] Take each component and mix them. Each component includes 35 mL of Ringer's solution, 150 mg of domperidone, and 0.15 mg of luteinizing hormone-releasing hormone A2, to obtain the oxytocic agent sample 2# for the large-scale reproduction of Bufo gargarizans.
[0048] Example 3
[0049] Take each component and mix them. Each component includes 28 mL of Ringer's solution, 150 mg of domperidone, and 0.15 mg of luteinizing hormone-releasing hormone A2, to obtain the oxytocic agent sample 3# for the large-scale reproduction of Bufo gargarizans.
[0050] Comparative Example 1
[0051] Take 30 mL of Ringer's solution to obtain the comparative oxytocic agent sample 1*.
[0052] Comparative Example 2
[0053] Take each component and mix them. Each component includes 30 mL of Ringer's solution, 3000 IU of chorionic gonadotropin, and 0.15 mg of luteinizing hormone-releasing hormone A2, to obtain the comparative oxytocic agent sample 2*.
[0054] Comparative Example 3
[0055] Take 15 mg, 30 mg, 90 mg, 300 mg, and 600 mg of domperidone respectively, and mix them with 30 mL of Ringer's solution and 0.15 mg of luteinizing hormone-releasing hormone A2 respectively to obtain the comparative oxytocic agent samples 3*-7*.
[0056] Comparative Example 4
[0057] Take 15 μg, 30 μg, 90 μg, 300 μg, and 600 μg of luteinizing hormone-releasing hormone A2 respectively, and mix them with 30 mL of Ringer's solution and 150 mg of domperidone respectively to obtain the comparative oxytocic samples 8*-12*.
[0058] Example 4
[0059] Select well-developed female and male Chinese giant toads as breeding toads. Among them, for female Chinese giant toads, it is required that the gonads are well-developed, the abdomen is enlarged, and there is a granular feeling when touched by hand; for male Chinese giant toads, it is required that the individual is large, the gonads are well-developed, the physical condition is healthy, and the vitality is good.
[0060] Pick up the abdominal skin of the selected female and male Chinese giant toads with forceps, insert the needle at a 45-degree angle, and be careful not to pierce into the abdominal cavity. Subcutaneously inject 0.5 mL of the oxytocic sample 1# prepared in Example 1 into the abdomen of each female Chinese giant toad, and subcutaneously inject 0.25 mL of the oxytocic sample 1# prepared in Example 1 into the abdomen of each male Chinese giant toad.
[0061] The injected female and male Chinese giant toads as breeding toads are placed together in a plastic breeding box for breeding. The number ratio of female Chinese giant toads to male Chinese giant toads in the same breeding box is 1:1. The water depth in the breeding box is 15 cm, and 6 fresh willow branches are placed in the breeding box to facilitate the female toads to lay eggs. The female and male Chinese giant toads start to lay eggs 24 hours after being placed together in the breeding box and end at 48 hours. After reaching the effective time, put the fertilized egg ribbons into the breeding pond and incubate them at 18 °C for 14 days, and then the tadpole seedlings 1# can be hatched.
[0062] Example 5
[0063] Select well-developed female and male Chinese giant toads as breeding toads. Among them, for female Chinese giant toads, it is required that the gonads are well-developed, the abdomen is enlarged, and there is a granular feeling when touched by hand; for male Chinese giant toads, it is required that the individual is large, the gonads are well-developed, the physical condition is healthy, and the vitality is good.
[0064] Pick up the abdominal skin of the selected female and male Chinese giant toads with forceps, insert the needle at a 48-degree angle, and be careful not to pierce into the abdominal cavity. Subcutaneously inject 0.4 mL of the oxytocic sample 2# prepared in Example 2 into the abdomen of each female Chinese giant toad, and subcutaneously inject 0.2 mL of the oxytocic sample 2# prepared in Example 2 into the abdomen of each male Chinese giant toad.
[0065] After injection, the female and male adult Chinese giant toads were placed together in a plastic breeding box for reproduction. The ratio of female to male Chinese giant toads in the same breeding box was 1:1.2. The water depth in the breeding box was 12 cm, and 5 fresh willow branches were placed in the breeding box to facilitate egg-laying by female toads. Egg-laying started 24 hours after the female and male Chinese giant toads were placed together in the breeding box and ended at 42 hours. After reaching the effective time, the fertilized egg ribbons were placed in a breeding pond and hatched at 12 °C for 18 days, and then tadpole seedlings 2# could be hatched.
[0066] Example 6
[0067] Adult Chinese giant toads, including well-developed females and males, were selected. Among them, for female Chinese giant toads, it was required that their gonads were well-developed, their abdomens were swollen, and there was a granular feeling when touched by hand; for male Chinese giant toads, it was required that they were large in size, had well-developed gonads, were in good physical health, and had good vitality.
[0068] The selected adult female and male Chinese giant toads were picked up by forceps to hold the abdominal skin, and the needle was inserted at a 42-degree angle, taking care not to pierce into the abdominal cavity. Each female Chinese giant toad was subcutaneously injected with 0.6 mL of the oxytocic agent sample 3# prepared in Example 3, and each male Chinese giant toad was subcutaneously injected with 0.3 mL of the oxytocic agent sample 3# prepared in Example 3.
[0069] After injection, the female and male adult Chinese giant toads were placed together in a plastic breeding box for reproduction. The ratio of female to male Chinese giant toads in the same breeding box was 1:1.5. The water depth in the breeding box was 18 cm, and 8 fresh willow branches were placed in the breeding box to facilitate egg-laying by female toads. Egg-laying started 24 hours after the female and male Chinese giant toads were placed together in the breeding box and ended at 45 hours. After reaching the effective time, the fertilized egg ribbons were placed in a breeding pond and hatched at 15 °C for 16 days, and then tadpole seedlings 3# could be hatched.
[0070] Test Example 1
[0071] 1. Test process
[0072] An oxytocic reproduction experiment was carried out at the Bufo bufo gargarizans medicinal material base of Shanghai Hutchison Pharmaceutical Co., Ltd. in Weishan County, Jining City, Shandong Province. The test time was from February 25th, 2025 to March 1st, 2025. A total of 3 batches of oxytocic reproduction experiments were carried out, and a total of 355 pairs of adult female and male Chinese giant toads were induced to ovulate. The adult female and male Chinese giant toads in these 3 batches were used for large-scale reproduction production. The oxytocic agent sample 1# of Example 1 was taken, and oxytocic induction and hatching were carried out according to Example 4, and the evaluation indexes of oxytocic effect were statistically analyzed. The specific results are shown in Table 1 below.
[0073] Meanwhile, take the oxytocic agent sample 1# of Example 1, and use 40 pairs of female and male Chinese giant toads as breeding toads for oxytocin induction and hatching according to Example 4, and count the evaluation indexes of the oxytocin induction effect. The specific results are shown in Table 1 below.
[0074] In addition, take the comparative oxytocic agent sample 1* prepared in Comparative Example 1, and use 40 pairs of female and male Chinese giant toads as breeding toads for oxytocin induction and hatching according to Example 4, and count the evaluation indexes of the oxytocin induction effect. The specific results are shown in Table 1 below.
[0075] Furthermore, take the comparative oxytocic agent sample 2* prepared in Comparative Example 2, and use 40 pairs of female and male Chinese giant toads as breeding toads for oxytocin induction and hatching according to Example 4, and count the evaluation indexes of the oxytocin induction effect. The specific results are shown in Table 1 below.
[0076] 2. Evaluation indexes of oxytocin induction effect
[0077] During the temporary rearing period, carefully count the number of ovulating breeding toads in each batch and the number of dead breeding toads during the temporary rearing period, and calculate the oxytocin induction rate, concentrated ovulation rate and mortality rate of each batch of breeding toads.
[0078] Oxytocin induction rate (%) = (the number of female breeding toads ovulating during the temporary rearing period / the total number of female breeding toads induced by oxytocin) × 100%, which is used to represent the overall oxytocin induction effect of the oxytocic agent.
[0079] Concentrated ovulation rate (%) = (the number of female breeding toads ovulating within a certain 15-hour stage during the temporary rearing period / the total number of female breeding toads induced by oxytocin) × 100%. The higher the concentrated ovulation rate, the more concentrated the effect time and the better the oxytocin induction effect.
[0080] Mortality rate (%) = (the number of dead female and male breeding toads during the temporary rearing period / the total number of female and male breeding toads induced by oxytocin) × 100%, which is used to reflect the death situation of breeding toads during the temporary rearing period after oxytocin induction.
[0081] 3. Comparison of test results
[0082] As can be seen from Table 1, for the oxytocin induction experiment results of 40 pairs of female and male Chinese giant toads, it is found that after using the oxytocic agent sample 1#, the oxytocin induction rate reaches 95%, the concentrated ovulation rate is 87.5%, and the mortality rate is 0%. The oxytocin induction rate and concentrated ovulation rate are higher than those of using the comparative oxytocic agent sample 1* in Comparative Example 1 for oxytocin induction. The oxytocin induction experiment results of Comparative Example 1 show that spawning is not concentrated, the temporary rearing time is longer, 5 toads die, and the mortality rate is 12.5%, which is relatively high. The oxytocin induction experiment results of Comparative Example 2 show that compared with using the oxytocic agent sample 1#, the oxytocin induction rate and concentrated ovulation rate of using the comparative oxytocic agent sample 2* in Comparative Example 2 are both lower than those of Example 1, and the mortality rate during the temporary rearing period is higher than that of Example 1.
[0083] Moreover, the experimental results of the large-scale propagation and production of three batches showed that the spawning rate of Bufo gargarizans after induction in the three batches exceeded 95%, the centralized spawning rate was over 85%, and the mortality rate during the temporary rearing period was less than 5%. It can be seen that the results of large-scale propagation are consistent with the experimental results, with good repeatability, and can be popularized and applied in practice.
[0084] Table 1 Effect of Spawning Induction Test
[0085]
[0086] 4. Conclusion
[0087] The spawning inducer of the present invention has a good spawning induction effect in the large-scale production of Bufo gargarizans, with a spawning rate of over 95%, a centralized spawning rate of over 85%, and a mortality rate of less than 5% during the temporary rearing period of the breeding toads. It can be popularized and used in the artificial large-scale breeding of medicinal Bufo gargarizans. In actual work, trials and demonstrations have been carried out in many places such as the toad venom medicinal material base of Shanghai Hehuang Pharmaceutical Co., Ltd., Weishan County, Jining City, Shandong Province, and Shenqiu County, Zhoukou City, Henan Province, etc., realizing the artificial large-scale reproduction of Bufo gargarizans.
[0088] Test Example 2
[0089] Take the comparative spawning inducer samples 3*-7* prepared in Comparative Example 3, and use 40 pairs of breeding toads of female and male Bufo gargarizans for spawning induction and hatching according to Example 4. Statistically evaluate the spawning induction effect indicators according to Test Example 1, and compare with the spawning induction and hatching results of the spawning inducer sample 1# in Example 1. The specific results are shown in Table 2 below.
[0090] Take the comparative spawning inducer samples 8*-12* prepared in Comparative Example 4, and use 40 pairs of breeding toads of female and male Bufo gargarizans for spawning induction and hatching according to Example 4. Statistically evaluate the spawning induction effect indicators according to Test Example 1, and compare with the spawning induction and hatching results of the spawning inducer sample 1# in Example 1. The specific results are shown in Table 3 below.
[0091] Table 2
[0092]
[0093] Table 3
[0094]
[0095] As can be seen from Tables 2 and 3, the spawning induction and hatching results of the spawning inducer sample 1# in Example 1 are the best, the spawning induction and hatching results of the comparative spawning inducer samples 5*, 6*, 10*, 11* are also relatively good, the spawning induction and hatching results of the comparative spawning inducer samples 4*, 9* are the second but still relatively good, and the spawning induction and hatching results of the comparative spawning inducer samples 4*, 7*, 8*, 12* are relatively poor.
[0096] In summary, the oxytocic agent and its oxytocic method for large-scale breeding of Bufo gargarizans provided by the present invention improve the breeding efficiency of Bufo gargarizans, can achieve large-scale artificial breeding of Bufo gargarizans, effectively promote the development of the artificial large-scale breeding industry of medicinal Bufo gargarizans, and are of great significance in promoting the development of artificial large-scale breeding amphibian breeding technology and genetic breeding of medicinal Bufo gargarizans. Therefore, the present invention effectively overcomes various disadvantages in the prior art and has high industrial utilization value.
[0097] The above embodiments are only illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Any person familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes made by those with ordinary knowledge in the technical field without departing from the spirit and technical ideas disclosed by the present invention should still be covered by the claims of the present invention.
Claims
1. An oxytocic agent, comprising Ringer's solution, domperidone, and luteinizing hormone-releasing hormone A2.
2. The oxytocic agent according to claim 1, wherein, In the oxytocic agent, the volume ratio of the Ringer's solution to the mass of domperidone is 1.0:1 - 10, mL / mg.
3. The oxytocic agent according to claim 1, characterized in that, In the oxytocic agent, the volume ratio of the Ringer's solution to the mass of luteinizing hormone-releasing hormone A2 is 1.0:1 - 10, mL / μg.
4. A preparation method of an oxytocic agent, comprising: Mix the components according to the ratio of the oxytocic agent described in any one of claims 1 - 3 to provide the required oxytocic agent.
5. Use of the oxytocic agent described in any one of claims 1 - 3 in the artificial oxytocin induction of Bufo gargarizans.
6. An oxytocic method for large-scale breeding of Bufo gargarizans, comprising: Subcutaneously inject the oxytocic agent described in any one of claims 1 - 3 into the abdomen of Bufo gargarizans. Separate the female and male Bufo gargarizans and place them together in a breeding box for amplexus and spawning.
7. The oxytocic method for large-scale breeding of Bufo gargarizans according to claim 6, characterized in that, Including any one or more of the following conditions: A1) When performing the subcutaneous injection, use forceps to pick up the abdominal skin of Bufo gargarizans and insert the needle at an angle of 40 - 50 degrees; A2) When the Bufo gargarizans is female, the injection volume for each female Bufo gargarizans is 0.4 - 0.6 mL / individual; A3) When the Bufo gargarizans is male, the injection volume for each male Bufo gargarizans is 0.2 - 0.3 mL / individual; A4) The number ratio of female Bufo gargarizans to male Bufo gargarizans in the same breeding box is 1:1 - 2; A5) Add water to the breeding box, and the water depth in the breeding box is 10 - 30 cm; A6) Place fresh branches in the breeding box; A7) The spawning time is not less than 24 h after the female and male Bufo gargarizans are placed together in the breeding box.
8. The oxytocic method for large-scale reproduction of Bufo gargarizans according to claim 7, characterized in that, Including any one or more of the following conditions: A61) In item A6, the branches are selected from at least one of willow branches or peach branches; A62) In item A6, the amount of branches placed is 5 - 8 for every 40 pairs of female and male Bufo gargarizans; A71) In item A7, the spawning time is 24 - 48 h after the female and male Bufo gargarizans are placed together in the breeding box.
9. A hatching method for large-scale breeding of Bufo gargarizans, comprising: Spawn according to the oxytocin induction method for large-scale reproduction of Bufo gargarizans described in any one of claims 6 - 8, collect the egg ribbons and put them into a breeding pond for hatching to hatch tadpole seedlings.
10. The hatching method for large-scale breeding of Bufo gargarizans according to claim 9, characterized in that, The water temperature in the breeding pond is 12 - 18 °C; and / or, the hatching time is 10 - 20 days.