A method for preventing prolapse of the cloaca in artificially bred juvenile chinese toad

Abstract

This invention provides a feed additive for preventing prolapse of juvenile Toad (Bufo gargarizans) in artificial breeding and its preparation method. This invention also provides a wet pellet feed for preventing prolapse of juvenile Toad (Bufo gargarizans) in artificial breeding and its preparation method. This invention further provides the use of the above-mentioned feed additive and / or wet pellet feed in preventing prolapse of juvenile Toad (Bufo gargarizans). This invention also provides a method for preventing prolapse of juvenile Toad (Bufo gargarizans). The method for preventing prolapse of juvenile Toad (Bufo gargarizans) in artificial breeding provided by this invention improves the stress resistance, immunity, and antioxidant capacity of juvenile Toads, thereby improving the overall health of juvenile Toads in artificial breeding and effectively preventing prolapse.

Description

Technical Field

[0001] This invention belongs to the technical field of agricultural breeding, specifically the technical field of medicinal animal breeding, and relates to a method for preventing prolapse of the anus in artificially bred young Chinese giant toads. Background Technology

[0002] Modern research shows that toad venom possesses various pharmacological activities, including cardiotonic, antitumor, analgesic, and local anesthetic effects. The Chinese giant toad (Bufo bufo gargarizans) is one of the basal organisms of toad venom. Due to habitat degradation and improper utilization, the wild population of the Chinese giant toad has declined significantly.

[0003] The Chinese giant toad primarily hatches into tadpoles via egg-carrying larvae, which then metamorphose into toad nymphs. After emerging onto land, these nymphs are highly susceptible to prolapse due to overeating of large-mouthed insects or changes in their diet caused by transitioning from live insects to live feed. The incidence rate ranges from 40% to 70%. Affected nymphs become lethargic, often lying under vegetation near water, stop eating, gradually become emaciated, and eventually die. The spontaneous recovery rate is extremely low, and group treatment is very difficult, necessitating preventative measures. Summary of the Invention

[0004] In view of the shortcomings of the prior art described above, the purpose of this invention is to provide a method for preventing prolapse of the anus in artificially bred Chinese toad larvae, which can enhance the stress resistance of the larvae and effectively reduce the incidence of prolapse in the larvae.

[0005] To achieve the above and other related objectives, the first aspect of this invention provides a feed additive for preventing prolapse of juvenile Toads in artificial breeding, which is selected from any one or more combinations of feed additive A, feed additive B, or feed additive C.

[0006] The feed additive A, by weight, comprises the following components:

[0007] Allicin 0.5-3 parts;

[0008] Pray for 0.5-3 portions of spirit;

[0009] Eucommia ulmoides extract 0.1-2 parts;

[0010] Bile acids 0.5-3 parts;

[0011] Antibiotics 0.5-3 parts;

[0012] The feed additive B, by weight, comprises the following components:

[0013] Carotenoids 0.5-3 parts;

[0014] Pray for 0.5-3 portions of spirit;

[0015] Multivitamins 0.5-3 servings;

[0016] Lactic acid bacteria 0.5-3 parts;

[0017] The feed additive C, by weight, comprises the following components:

[0018] Carotenoids 0.5-3 parts;

[0019] Pray for 0.5-3 portions of spirit;

[0020] Multivitamins 0.5-3 servings;

[0021] 0.5-3 parts of Bacillus.

[0022] The second aspect of the present invention provides a method for preparing a feed additive for preventing prolapse of artificially bred juvenile Toads, comprising: mixing any one of the components of feed additive A, feed additive B or feed additive C in the above-mentioned feed additive for preventing prolapse of artificially bred juvenile Toads.

[0023] A third aspect of the present invention provides a wet pellet feed for preventing prolapse of artificially bred juvenile Chinese toads, comprising the aforementioned feed additive for preventing prolapse of artificially bred juvenile Chinese toads.

[0024] The fourth aspect of this invention provides a method for preparing wet pelleted feed for preventing prolapse of artificially bred Chinese giant toad larvae, comprising: adding feed additives for preventing prolapse of artificially bred Chinese giant toad larvae to pelleted feed and water according to a specified ratio, and mixing them evenly.

[0025] The fifth aspect of the present invention provides the above-mentioned feed additive for preventing prolapse of artificially bred Toad larvae, and / or the use of the above-mentioned wet pellet feed for preventing prolapse of artificially bred Toad larvae in preventing prolapse of artificially bred Toad larvae.

[0026] The sixth aspect of the present invention provides a method for preventing prolapse of artificially bred juvenile Chinese toads, comprising: first feeding the newly opened juvenile Chinese toads with a mixture of live insect bait and the aforementioned wet pellet feed in a first stage, and then feeding them with the aforementioned wet pellet feed in a second stage.

[0027] As described above, the method for preventing prolapse of the anus in artificially bred young Chinese giant toads provided by the present invention has the following beneficial effects:

[0028] (1) After young toads come ashore, they are prone to prolapse when switching from live larvae to live feed, especially when they consume larvae of excessively large size or when their diet changes. Treatment of prolapsed young toads is difficult, necessitating preventative measures. This invention provides a method for preventing prolapse in artificially bred young Toads of the Chinese Giant Toad (Bufo gargarizans). By feeding young toads appropriately sized larvae, preparing wet pellet feed, and using feed additives, the method enhances the young toads' stress resistance, immunity, and antioxidant capacity, effectively preventing prolapse.

[0029] (2) The present invention provides a method for preventing prolapse of artificially bred Toad larvae. The method determines the appropriate size of the insects the larvae should ingest. Newly hatched larvae should feed on live insects approximately 3-5 mm long and 0.5 mm in diameter. When the larvae weigh more than 1.0 g, they should feed on live insects approximately 5-8 mm long and 1.0 mm in diameter. Feeding larvae live insects of the above specifications effectively prevents prolapse caused by ingesting excessively large live insects.

[0030] (3) Since live insects have a water content of 60-70%, while commercially available pelleted feed generally has a water content of around 10%, the diet of young toads changes during the transition from live insects to fresh feed, making them prone to stress-induced enteritis and ultimately leading to prolapse. This invention provides a method for preventing prolapse in artificially bred young Toads of the Chinese Giant Toad. The optimal water-to-feed ratio was determined through experimentation. When mixing the feed, tap water of approximately 30% of the feed weight is added to increase the water content of the pelleted feed, softening it. After the young toads consume the wet pelleted feed, their digestibility and absorption rate improves, further preventing prolapse.

[0031] (4) The present invention provides a method for preventing prolapse of artificially bred Chinese toad larvae. It uniquely adopts multiple wet pellet feed formulas and feeds them in rotation. In multiple feed additive formulas, it adds basil to improve the stress resistance of larvae and prevent stress enteritis during the transition period; Eucommia ulmoides extract enhances the immunity of larvae; carotenoids improve the antioxidant stress resistance of larvae; probiotics improve the intestinal flora of larvae, and improve the physical condition of artificially bred Chinese toad larvae from all aspects, effectively reducing the prolapse rate of larvae. Attached Figure Description

[0032] Figure 1 The image shown is a diagram of the individual state of a young Chinese giant toad with prolapsed rectum, as shown in Comparative Example 1.

[0033] Figure 2 The diagram shows the group status of young Chinese giant toads with rectal prolapse, as shown in Comparative Example 1.

[0034] Figure 3 The diagram shows the population state of young toads feeding on 3-5 mm open-mouthed insects in Example 5.

[0035] Figure 4The diagram shows the population state of young toads feeding on 5-8 mm open-mouthed insects in Example 5.

[0036] Figure 5 The diagram shows the population state of juvenile toads in Example 5, who were fed a mixture of open-mouth insects and wet pelleted feed containing feed additives.

[0037] Figure 6 The image shown is a group diagram of the Chinese giant toad larvae growing to 10.0g in Example 5.

[0038] Figure 7 The graph shows the effect of feed additives on the prolapse rate of juvenile toads in comparative test example 2.

[0039] Figure 8 The diagram shows the relative abundance of gut microbiota genera in juvenile toads in comparative test example 2, where H_BUFO1 represents the control group toads and H_BUFO2 represents the experimental group toads. Detailed Implementation

[0040] The first aspect of this invention provides a feed additive for preventing prolapse of juvenile Toads in artificial breeding, comprising any one or more combinations selected from feed additive A, feed additive B, or feed additive C.

[0041] The feed additive A, by weight, comprises the following components:

[0042] Allicin: 0.5-3 parts; specifically, 0.5-1 part, 1-2 parts, or 2-3 parts.

[0043] Bai Ji Ling: 0.5-3 portions; specifically, 0.5-1 portion, 1-2 portions, or 2-3 portions.

[0044] Eucommia ulmoides extract, 0.1-2 parts; specifically, 0.1-0.5 parts, 0.5-1 part, or 1-2 parts;

[0045] Bile acids 0.5-3 parts; specifically 0.5-1 part, 1-2 parts, 2-3 parts;

[0046] Antibiotics: 0.5-3 parts; specifically, 0.5-1 part, 1-2 parts, or 2-3 parts.

[0047] The feed additive B, by weight, comprises the following components:

[0048] Carotenoids: 0.5-3 parts; specifically, 0.5-1 part, 1-2 parts, or 2-3 parts.

[0049] Bai Ji Ling: 0.5-3 portions; specifically, 0.5-1 portion, 1-2 portions, or 2-3 portions.

[0050] Multivitamins: 0.5-3 servings; specifically, 0.5-1 serving, 1-2 servings, or 2-3 servings.

[0051] Lactic acid bacteria: 0.5-3 parts; specifically, 0.5-1 part, 1-2 parts, or 2-3 parts.

[0052] The feed additive C, by weight, comprises the following components:

[0053] Carotenoids: 0.5-3 parts; specifically, 0.5-1 part, 1-2 parts, or 2-3 parts.

[0054] Bai Ji Ling: 0.5-3 portions; specifically, 0.5-1 portion, 1-2 portions, or 2-3 portions.

[0055] Multivitamins: 0.5-3 servings; specifically, 0.5-1 serving, 1-2 servings, or 2-3 servings.

[0056] Bacillus subtilis 0.5-3 parts; specifically, 0.5-1 part, 1-2 parts, or 2-3 parts.

[0057] In one embodiment, the feed additive A comprises, by weight, the following components:

[0058] Allicin 1-2 parts;

[0059] Pray to the spirits 1-2 times;

[0060] Eucommia ulmoides extract 0.5-1 part;

[0061] Bile acids 1-2 parts;

[0062] 1-2 doses of antibiotics;

[0063] The feed additive B, by weight, comprises the following components:

[0064] Carotenoids 1-2 parts;

[0065] Pray to the spirits 1-2 times;

[0066] 1-2 servings of multivitamins;

[0067] 1-2 servings of lactic acid bacteria;

[0068] The feed additive C, by weight, comprises the following components:

[0069] Carotenoids 1-2 parts;

[0070] Pray to the spirits 1-2 times;

[0071] 1-2 servings of multivitamins;

[0072] 1-2 portions of Bacillus.

[0073] In one embodiment, the above allicin, Baijiling, Eucommia ulmoides extract, bile acid, antibiotic, carotenoid, compound vitamin, lactic acid bacteria, and bacillus are all corresponding components commonly used and can be obtained by purchasing from the market.

[0074] In a preferred embodiment, the allicin is purchased from Hebi Xingwang Biotechnology Co., Ltd., production license number: Yu Si Tian (2019) H06174.

[0075] In a preferred embodiment, the Baijiling is purchased from Elanco (Sichuan) Animal Health Co., Ltd., production license number: Chuan Si Yu (2020) 01013.

[0076] In a preferred embodiment, the Eucommia ulmoides extract is purchased from Anhui Best Biotechnology Co., Ltd., production license number: Wan Si Tian (2023) H01008.

[0077] In a preferred embodiment, the bile acid is purchased from Henan Jiuchu Biotechnology Co., Ltd., production license number: Yu Si Tian (2021) H01315.

[0078] In a preferred embodiment, the antibiotic is gentamicin sulfate, with a CAS number of 1405 - 41 - 0. Specifically, the antibiotic is gentamicin sulfate soluble powder, purchased from Jiangsu Blue Whale Longma Biotechnology Co., Ltd., production license number: (2020) Veterinary Drug Production License No. 1016601013.

[0079] In a preferred embodiment, the carotenoid is purchased from Hubei Xinhe Biotechnology Co., Ltd., production license number: SC20142120100012.

[0080] In a preferred embodiment, the compound vitamin is purchased from Guangzhou Jinshui Animal Health Products Co., Ltd., production license number: Yue Si Yu (2020) 01062.

[0081] In a preferred embodiment, the lactic acid bacteria are purchased from Henan Nanhua Qianmu Biotechnology Co., Ltd., production license number: Yu Si Yu (2019) H0709.

[0082] In a preferred embodiment, the bacillus is purchased from Guangzhou Jinshui Animal Health Products Co., Ltd., production license number: Shou Si Tian (2019) H010118.

[0083] The second aspect of the present invention provides a preparation method for a feed additive for preventing prolapse of the anus in artificially - cultured juvenile Bufo gargarizans, including: mixing the components of any one of feed additive A, feed additive B, or feed additive C in the above feed additive for preventing prolapse of the anus in artificially - cultured juvenile Bufo gargarizans to obtain it.

[0084] In one embodiment, the mixing is carried out using a feed mixer.

[0085] A third aspect of the present invention provides a wet pellet feed for preventing prolapse of artificially bred juvenile Chinese toads, comprising the aforementioned feed additive for preventing prolapse of artificially bred juvenile Chinese toads.

[0086] In one embodiment, the wet pellet feed for preventing prolapse of juvenile Toads in artificial breeding comprises the following components by weight:

[0087] 1000 portions of pelleted feed;

[0088] Water 200-400 parts; specifically, 200-250 parts, 250-350 parts, 350-400 parts, preferably 300 parts;

[0089] The feed additive for preventing prolapse of the anus in artificially bred juvenile Chinese toads is selected from any one or more combinations of feed additive A, feed additive B, or feed additive C. Specifically, feed additive A is 2.1-14 parts, such as 2.1-4.5 parts, 4.5-9 parts, or 9-14 parts, preferably 4.5-9 parts; feed additive B is 2-12 parts, such as 2-4 parts, 4-8 parts, or 8-12 parts, preferably 4-8 parts; and feed additive C is 2-12 parts, such as 2-4 parts, 4-8 parts, or 8-12 parts, preferably 4-8 parts.

[0090] In a preferred embodiment, the pelleted feed is a conventionally used extruded compound feed for frogs. Specifically, the pelleted feed is an extruded compound feed for black-spotted frogs, purchased from Yangzhou Hongda Feed Co., Ltd., with production license number: Su Si Zheng (2020) 10003.

[0091] In a preferred embodiment, the particle size of the pelleted feed is 0.8-5.0 mm, specifically such as 0.8-1.0 mm, 1.0-1.7 mm, 1.7-3.3 mm, 3.3-5.0 mm, and preferably such as 0.8 mm, 1.0 mm, 1.7 mm, 3.3 mm, 5.0 mm.

[0092] In a preferred embodiment, the water is tap water.

[0093] The fourth aspect of this invention provides a method for preparing wet pelleted feed for preventing prolapse of artificially bred Chinese giant toad larvae, comprising: adding feed additives for preventing prolapse of artificially bred Chinese giant toad larvae to pelleted feed and water according to a specified ratio, and mixing them evenly.

[0094] In one embodiment, the mixing is carried out using a feed mixer.

[0095] The fifth aspect of the present invention provides the above-mentioned feed additive for preventing prolapse of artificially bred Toad larvae, and / or the use of the above-mentioned wet pellet feed for preventing prolapse of artificially bred Toad larvae in preventing prolapse of artificially bred Toad larvae.

[0096] The sixth aspect of the present invention provides a method for preventing prolapse of artificially bred juvenile Chinese toads, comprising: first feeding the newly opened juvenile Chinese toads with a mixture of live insect bait and the aforementioned wet pellet feed in a first stage, and then feeding them with the aforementioned wet pellet feed in a second stage.

[0097] In one embodiment, the larvae are obtained by taking selected larvae of the Chinese giant toad and feeding them live insects during the larval stage to induce them to open their mouths.

[0098] In a preferred embodiment, the weight of the juvenile toad is 0.2-0.5g per toad. The preferred juvenile toads are those that have just come ashore.

[0099] In a preferred embodiment, the live insect bait is a live insect commonly used to feed young toads, specifically including, but not limited to, live mealworms, live black soldier flies, and live fly larvae.

[0100] In a preferred embodiment, the live insects in the live insect bait are 3-5 mm in length and 0.4-0.6 mm in diameter, preferably 0.5 mm.

[0101] In a preferred embodiment, the daily feeding amount of the live insect bait is 9-11 wt% of the body weight of the juvenile toad that has come ashore, preferably 10 wt%.

[0102] In a preferred embodiment, the live insect bait is fed at 5-7 a.m. and 5-7 p.m. daily, preferably at 6 a.m. and 6 p.m. daily.

[0103] In a preferred embodiment, the live insect bait is fed 2-3 times per day, preferably 2 times per day.

[0104] The opening period refers to the time from when the young toads come ashore until their average weight increases to 1g.

[0105] In one embodiment, the first stage of feeding and / or the second stage of feeding are carried out on a feeding platform. The feeding platform is a conventional toad feeding platform equipped with a vibrator to keep the feed vibrating. The feeding platform is kept clean, and any remaining feed is promptly removed.

[0106] In one embodiment, during the first stage of feeding, the daily amount of the live insect bait and the aforementioned wet pellet feed is 9-11 wt% of the body weight of the newly hatched toad, preferably 10 wt%.

[0107] In one embodiment, the feeding times for the first stage of feeding and / or the second stage of feeding are 5-7 a.m. and 5-7 p.m. daily, preferably 6 a.m. and 6 p.m. daily.

[0108] In a preferred embodiment, the amount of feed given in the morning is 25-35% of the daily feed amount, preferably 30%; and the amount of feed given in the afternoon is 65-75% of the daily feed amount, preferably 70%.

[0109] In a preferred embodiment, the feeding frequency of the first stage feeding and / or the second stage feeding is 2-3 times / day, preferably 2 times / day.

[0110] In one embodiment, during the first stage of feeding, with the total amount of live insect bait and wet pellet feed fixed at 100 wt%, on day 1, the mixing ratio of live insect bait and wet pellet feed is 9:1 (i.e., the amount of live insect bait is 90 wt% and the amount of wet pellet feed is 10 wt%); from day 2 to day 9-19, preferably day 9-14, the amount of pellet feed is increased by 10 wt% for 1-2 days, changing to a mixing ratio of live insect bait and wet pellet feed of 1:9 (i.e., the amount of live insect bait is 10 wt% and the amount of wet pellet feed is 90 wt%).

[0111] In one embodiment, during the second stage of feeding, the daily amount of the wet pellet feed is 9-11 wt% of the body weight of the newly hatched toad, preferably 10 wt%.

[0112] In one embodiment, during the second stage of feeding, from the first day after the end of the first stage of feeding until the average weight of the juvenile toads reaches 10g, the feeding uses only wet pellet feed (i.e., the amount of live insect bait is 0wt%, and the amount of wet pellet feed is 100wt%).

[0113] In one embodiment, during the first stage of feeding, the live insects in the live insect bait are 5-8 mm in length and 0.9-1.1 mm in diameter, preferably 1.0 mm.

[0114] In one embodiment, during the first stage of feeding and / or the second stage of feeding, the feed additive in the wet pellet feed is feed additive A during the first stage; during the second stage, the feed additive in the wet pellet feed is feed additive B during the second stage; during the third stage, the feed additive in the wet pellet feed is feed additive C; and during the subsequent stages, the feed additives are added to the wet pellet feed in the above order in a cyclical manner.

[0115] In a preferred embodiment, the stage duration is not less than 3 days, preferably 3 days. The stage duration includes, but is not limited to, the first stage duration, the second stage duration, the third stage duration, and the subsequent stage duration.

[0116] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of protection of the present invention.

[0117] The following specific examples illustrate the implementation 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 embodiments, and 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.

[0118] Example 1

[0119] Feed additive sample A (sample 1) was prepared by mixing 2g allicin, 2g acarbose, 1g eucommia extract, 2g bile acids, and 2g gentamicin sulfate in a feed mixer. Feed additive sample B (sample 1) was prepared by mixing 2g carotenoids, 2g acarbose, 2g multivitamins, and 2g lactic acid bacteria in a feed mixer. Feed additive sample C (sample 1) was prepared by mixing 2g carotenoids, 2g acarbose, 2g multivitamins, and 2g Bacillus subtilis in a feed mixer. Thus, feed additive sample 1 was obtained.

[0120] Example 2

[0121] Feed additive sample A (sample 2) was prepared by mixing 1g allicin, 1g acarbose, 0.5g eucommia extract, 1g bile acid, and 1g gentamicin sulfate in a feed mixer. Feed additive sample B (sample 2) was prepared by mixing 1g carotenoids, 1g acarbose, 1g multivitamins, and 1g lactic acid bacteria in a feed mixer. Feed additive sample C (sample 2) was prepared by mixing 1g carotenoids, 1g acarbose, 1g multivitamins, and 1g Bacillus subtilis in a feed mixer. Thus, feed additive sample 2 was obtained.

[0122] Example 3

[0123] Take 1000g of 0.8mm pelleted feed and 300g of tap water, add 9g of feed additive A sample 1 prepared in Example 1, and mix evenly in a feed mixer to obtain wet pelleted feed sample A1. Take 1000g of 0.8mm pelleted feed and 300g of tap water, add 8g of feed additive B sample 1 prepared in Example 1, and mix evenly in a feed mixer to obtain wet pelleted feed sample B1. Take 1000g of 0.8mm pelleted feed and 300g of tap water, add 8g of feed additive C sample 1 prepared in Example 1, and mix evenly in a feed mixer to obtain wet pelleted feed sample C1. Thus, wet pelleted feed sample 1# is obtained.

[0124] Example 4

[0125] Take 1000g of 0.8mm pelleted feed and 300g of tap water, add 4.5g of feed additive A sample 2 prepared in Example 2, and mix evenly in a feed mixer to obtain wet pelleted feed sample A2. Take 1000g of 0.8mm pelleted feed and 300g of tap water, add 4g of feed additive B sample 2 prepared in Example 2, and mix evenly in a feed mixer to obtain wet pelleted feed sample B2. Take 1000g of 0.8mm pelleted feed and 300g of tap water, add 4g of feed additive C sample 2 prepared in Example 2, and mix evenly in a feed mixer to obtain wet pelleted feed sample C2. Thus, wet pelleted feed sample 2# is obtained.

[0126] Example 5

[0127] Selected juvenile Chinese toads, weighing 0.2-0.5g each, were fed live insects during their initial feeding period to induce them to start eating, resulting in juveniles weighing 1.0g. The live insects used were live mealworms, such as... Figure 3 As shown, the live insects were 4 mm long and 0.5 mm in diameter. The daily feeding amount of live insects was 10 wt% of the juvenile toad's body weight, and the feeding times were 6:00 AM and 6:00 PM, twice a day.

[0128] After reaching a weight of 1.0g, the newly hatched toads were first fed a mixture of live insect bait and wet pellet feed sample 1# prepared in Example 3, followed by a second stage of feeding with the same wet pellet feed sample 1# prepared in Example 3. The live insect bait consisted of live mealworms, such as... Figure 4 As shown, the length of the live insect bait was 7 mm; the diameter of the live insect was 1.0 mm. In the first stage of feeding, the daily feeding amount of live insect bait and wet pellet feed sample 1# was 10 wt% of the body weight of the newly hatched toads; in the second stage of feeding, the daily feeding amount of wet pellet feed sample 1# was 10 wt% of the body weight of the newly hatched toads.

[0129] In both the first and second stages of feeding, feeding was conducted on a feeding platform at 6:00 AM and 6:00 PM daily. The morning feeding amount was 30% of the daily total, and the afternoon feeding amount was 70% of the daily total. From the start of feeding, for days 1-3, wet pellet feed sample #1 was mixed with wet pellet feed sample A1 for three days; for days 4-6, wet pellet feed sample #1 was mixed with wet pellet feed sample B1 for three days; for days 7-9, wet pellet feed sample #1 was mixed with wet pellet feed sample C1 for three days, and so on, in a cyclical manner.

[0130] In the first stage of feeding, such as Figure 5 As shown, with the total amount of live insect feed and wet pellet feed fixed at 100 wt%, on day 1, the amount of live insect feed was 90 wt% and the amount of wet pellet feed was 10 wt%. From day 2 to day 10, the amount of pellet feed was increased by 10 wt% per day, changing to a total amount of live insect feed of 10 wt% and wet pellet feed of 90 wt%.

[0131] In the second feeding phase, from the day after the end of the first feeding phase until the juvenile toads reached an average weight of 10g, only wet pellet feed was used. Specifically, the amount of live insect feed was 0wt%, and the amount of wet pellet feed was 100wt%. Figure 6 As shown, the final sample of juvenile toads with an average weight of 10g was obtained.

[0132] Example 6

[0133] Young Chinese toads, weighing 0.2-0.5g each, were fed live insects during their initial feeding period to induce them to start eating, resulting in juveniles weighing 1.0g. The live insects used were live black soldier flies, with a length of 5mm and a diameter of 0.6mm. The daily feeding amount of live insects was 10.5wt% of the juvenile toad's body weight, and the feeding times were 6:00 AM and 6:00 PM, twice a day.

[0134] After reaching a weight of 1.0g, the newly hatched toad larvae were first fed a mixture of live insect bait and wet pellet feed sample 2# prepared in Example 4, followed by a second stage of feeding with the same wet pellet feed sample 2#. The live insect bait consisted of live black soldier flies, each 6mm in length and 0.9mm in diameter. During the first stage, the daily amount of live insect bait and wet pellet feed sample 2# was 10.5wt% of the larvae's body weight; during the second stage, the daily amount of wet pellet feed sample 2# was also 10.5wt% of the larvae's body weight.

[0135] In both the first and second phases of feeding, feeding was conducted on a feeding platform at 6:00 AM and 6:00 PM daily. The morning feeding amount was 28% of the daily total, and the afternoon feeding amount was 72% of the daily total. From the start of feeding, for days 1-3, wet pellet feed sample 2# was mixed with wet pellet feed sample A2 for three days; for days 4-6, wet pellet feed sample 2# was mixed with wet pellet feed sample B2 for three days; for days 7-9, wet pellet feed sample 2# was mixed with wet pellet feed sample C2 for three days, and so on, in a cyclical manner.

[0136] In the first stage of feeding, with the total amount of live insect feed and wet pellet feed fixed at 100 wt%, on day 1, the amount of live insect feed was 90 wt% and the amount of wet pellet feed was 10 wt%. From day 2 to day 10, the amount of pellet feed was increased by 10 wt% per day, until the amount of live insect feed was 10 wt% and the amount of wet pellet feed was 90 wt%.

[0137] In the second feeding phase, from the day after the end of the first feeding phase until the average weight of the juvenile toads reached 10g, only wet pellet feed was used, i.e., the amount of live insect bait was 0wt%, and the amount of wet pellet feed was 100wt%. Finally, juvenile toad sample 2# with an average weight of 10g was obtained.

[0138] Comparative Example 1

[0139] After selecting 0.2-0.5g juvenile Chinese toads and feeding them using conventional methods for two weeks, it was observed that the juveniles experienced rectal prolapse, sluggish movement, often lying under waterside vegetation, ceasing to eat, gradually becoming emaciated, and eventually dying. Figure 1 As shown, the individual characteristics of prolapsed toad larvae can be observed, such as... Figure 2 As shown, the group status of prolapsed toad larvae can be observed.

[0140] Comparison Test Example 1

[0141] Two breeding ponds were selected, divided into an experimental group and a control group. The initial weight of the juvenile toads in both groups was 1.0g, and the experiment lasted for 40 days. The experimental group was fed two stages of different feeds according to Example 5, while the control group was fed only with feed additive sample 1 prepared in Example 1. Other feeding conditions were the same as in Example 5. The experimental results are shown in Table 1.

[0142] Table 1. Data on juvenile toad rearing in different embodiments

[0143] experimental group control group Rectal prolapse rate (%) 1.2 83 Daily weight gain (%) 1.09 1.07 Time (days) to reach 10g 27 34

[0144] As shown in Table 1, the prolapse rate of the juvenile toads fed in Example 5 was lower than that in Example 1, and the daily weight gain rate was higher than that in Example 1. It took 27 days for the juvenile toads in Example 5 to grow to 10g, which was faster than the 34 days required for the juvenile toads in Example 1 to grow to 10g.

[0145] Comparison Test Example 2

[0146] Two breeding ponds were selected, divided into an experimental group and a control group. The initial weight of the juvenile toads in both groups was 1.0g, and the experiment lasted for 40 days. The experimental group was fed two stages of different feeds as described in Example 5, while the control group was fed only 30% of the feed weight of tap water-soaked wet pellets without feed additives. Other feeding conditions were the same as in Example 5. The experimental results are shown below. Figure 7 and Figure 8 .

[0147] Depend on Figure 7 It can be seen that the prolapse rate in the experimental group was 1.2%, while the prolapse rate in the control group was 47.0%. The prolapse rate of the young toads fed using Example 5 was lower than that in the control group.

[0148] Depend on Figure 8 Analysis of the small intestinal flora in both the experimental and control groups revealed that the top four dominant genera at the genus level were *Citrobacter*, *Bacteroides*, *Faecalicatena*, and *Desulfovibrio*. The abundance of *Citrobacter*, a potentially pathogenic bacterium, was higher in the control group than in the experimental group, indicating that young toads in the control group were more susceptible to bacterial infection.

[0149] Comparison Test Case 3

[0150] Three breeding ponds were selected and divided into an experimental group, a control group 1, and a control group 2. The initial weight of the juvenile toads in all three groups was 1.0g, and the experiment lasted for 40 days. The experimental group was fed the same as in comparative test example 1, but with two different feeds in two stages according to Example 5 (tap water accounted for 30% of the feed weight in wet pellet feed sample 1#). Control group 1 was fed wet pellet feed containing 30% tap water by weight without feed additives, and control group 2 was fed wet pellet feed containing 10% tap water by weight without feed additives. Other feeding conditions were the same as in Example 5. The experimental results are shown in Table 2.

[0151] Table 2. Feeding juvenile toads with different proportions of wet pelleted feed

[0152] experimental group Control group 1 Control group 2 Rectal prolapse rate (%) 1.2 21 64 Daily weight gain (%) 1.09 1.07 1.06 Time (days) to reach 10g 27 34 40

[0153] As shown in Table 2, the prolapse rate of the experimental group was lower than that of the control groups 1 and 2, and the daily weight gain rate was higher than that of the control groups 1 and 2. The experimental group's toads only needed 27 days to grow to 10g, which was faster than the 34 days required for the toads in control group 1 to grow to 10g and the 40 days required for the toads in control group 2 to grow to 10g.

[0154] In addition, the prolapse rate of juvenile toads fed with wet pelleted feed containing 30% tap water by weight in control group 1 was lower than that in control group 2 fed with 10% tap water by weight. The daily weight gain rate of control group 1 was slightly higher than that of control group 2 fed with 10% tap water by weight. It took 34 days for juvenile toads in control group 1 fed with 30% tap water by weight to grow to 10g, which was faster than the 40 days required for juvenile toads in control group 2 fed with 10% tap water by weight to grow to 10g.

[0155] In summary, the present invention provides a method for preventing prolapse of artificially bred Toad larvae, improving their stress resistance, immunity, and antioxidant capacity, thereby enhancing their overall health and effectively preventing prolapse. Therefore, this invention effectively overcomes the shortcomings of existing technologies and possesses high industrial application value.

[0156] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

Claims

1. A feed additive, comprising any one or more combinations selected from feed additive A, feed additive B, or feed additive C. The feed additive A, by weight, comprises the following components: Allicin 0.5-3 parts; Pray for 0.5-3 portions of the spirit; Eucommia ulmoides extract 0.1-2 parts; Bile acids 0.5-3 parts; Antibiotics 0.5-3 parts; The feed additive B, by weight, comprises the following components: Carotenoids 0.5-3 parts; Pray for 0.5-3 portions of spirit; Multivitamins 0.5-3 servings; Lactic acid bacteria 0.5-3 parts; The feed additive C, by weight, comprises the following components: Carotenoids 0.5-3 parts; Pray for 0.5-3 portions of spirit; Multivitamins 0.5-3 servings; 0.5-3 parts of Bacillus.

2. A method of preparing a feed additive comprising: The feed additive described in claim 1 is prepared by mixing any one of the components of feed additive A, feed additive B, or feed additive C.

3. A wet pelleted feed comprising the feed additive of claim 1.

4. The wet-pelleted feed according to claim 3, characterized in that, The wet pelleted feed, by weight, comprises the following components: 1000 portions of pelleted feed; 200-400 parts water; The feed additive is selected from any one or more combinations of feed additive A, feed additive B, or feed additive C, wherein feed additive A is 2.1-14 parts; feed additive B is 2-12 parts; and feed additive C is 2-12 parts.

5. A method of preparing a wet pelleted feed comprising: The feed additive described in claim 1 is added to the pelleted feed and water according to the specified ratio and mixed evenly.

6. The use of the feed additive of claim 1, and / or the wet pellet feed of any one of claims 3 or 4 in preventing prolapse of juvenile Toads in artificial breeding.

7. A method for preventing prolapse of the cloaca in artificially bred Bufo bufo gargarizans tadpoles, comprising: The young toads of the Chinese giant toad were first fed a mixture of live insect bait and wet pellet feed as described in claim 3 or 4 in the first stage, and then fed a second stage of wet pellet feed as described in claim 3 or 4.

8. The method for preventing proctedysis of the artificial breeding Bufo bufovar. gargarizans tadpole according to claim 7, characterized in that, The aforementioned juvenile toads are obtained by taking selected juvenile toads of the Chinese giant toad and feeding them live insects during the opening period to induce them to open their mouths.

9. The method for preventing proctedysis of the artificial breeding Bufo bufovar. gargarizans tadpole according to claim 8, characterized in that, Includes one or more of the following conditions: A1) The weight of the juvenile toad is 0.2-0.5g / toad; A2) The live insect bait is selected from at least one of live mealworms, live black soldier flies, and live fly larvae. A3) The live insects in the live insect bait have a length of 3-5 mm and a diameter of 0.4-0.6 mm; A4) The daily feeding amount of the live insect bait is 9-11 wt% of the body weight of the juvenile toads that have come ashore; A5) The feeding time for the live insect bait is from 5:00 AM to 7:00 AM and from 5:00 PM to 7:00 PM every day.

10. The method for preventing prolapse of the cloaca in juvenile artificial breeding Chinese toad according to claim 7, characterized in that, Includes one or more of the following conditions: B1) During the first stage of feeding, the daily feeding amount of the live insect bait and the above-mentioned wet pellet feed is 9-11 wt% of the body weight of the newly opened toad larvae; B2) The feeding times for the first stage of feeding and / or the second stage of feeding are 5-7 am and 5-7 pm every day; preferably, the amount of feeding in the morning is 25-35% of the daily feeding amount; and the amount of feeding in the afternoon is 65-75% of the daily feeding amount. B3) In the first stage of feeding, with the total amount of live insect bait and wet pellet feed fixed at 100 wt%, on day 1, the mixing ratio of live insect bait and wet pellet feed is 9:1; from day 2 to day 9-19, the amount of pellet feed is increased by 10 wt% every 1-2 days, changing to a mixing ratio of live insect bait and wet pellet feed of 1:9; B4) In the second stage of feeding, the daily amount of wet pellet feed is 9-11 wt% of the body weight of the newly hatched toads; B5) In the second stage of feeding, that is, from the first day after the end of the first stage of feeding until the average weight of the young toads increases to 10g, only wet pellet feed is used for feeding. B6) In the first stage of feeding, the live insects in the live insect bait are 5-8 mm in length and 0.9-1.1 mm in diameter; B7) In the first stage of feeding and / or the second stage of feeding, during the first stage, the feed additive in the wet pellet feed is feed additive A; during the second stage, the feed additive in the wet pellet feed is feed additive B; during the third stage, the feed additive in the wet pellet feed is feed additive C; during the subsequent stages, the feed additives are added to the wet pellet feed in the above order in a cyclical manner; preferably, the stage time is not less than 3 days.

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