Thermoregulatory animal feed, method for its preparation and use

Abstract

The present application belongs to the technical field of feed, and particularly relates to a feed for poikilothermic animals, a preparation method and application thereof. The feed for poikilothermic animals has a core and a shell layer arranged on the surface of the core. The core comprises sodium alginate, a protein base, an attractant A and a fermentation product synergist. The preparation raw materials of the shell layer comprise sodium alginate and calcium lactate. The shell layer comprises an attractant B. The attractant A comprises an insect-derived attractant, an animal-derived attractant and a plant-derived attractant. The attractant B comprises a freeze-dried powder containing fructose and an insect enzymatic hydrolysate. Through the synergistic effect of the insect-derived attractant, the animal-derived attractant and the plant-derived attractant, the umami synergistic effect of the fermentation product synergist and the insect-derived attractant in the attractant A, and the preparation process combining low temperature and irradiation sterilization technology, the feed for poikilothermic animals has good feeding attraction, nutrient retention rate and safety.

Description

Technical Field

[0001] This invention belongs to the field of feed technology, and specifically relates to a cold-blooded animal feed, its preparation method, and its application. Background Technology

[0002] In recent years, with the improvement of people's living standards, the pet industry, which focuses on keeping cold-blooded animals (including fish, reptiles, amphibians, and other animal groups with weak thermoregulation capabilities), has flourished. From ornamental fish (such as arowanas, tigerfish, and podocarpus), amphibians (such as horned frogs and axolotls) to reptiles (such as geckos, lizards, and corn snakes), these pets have become an important part of the pet market due to their unique ecological habits, low space requirements, and diverse ornamental value. The corresponding pet food needs to adapt to the different dietary differences of these groups—fish rely on plankton, algae, or small aquatic animals; amphibians mainly prey on insects or small fish and shrimp; and reptiles include insectivorous, herbivorous, and carnivorous (including fish and rodents) types, posing diverse requirements for the nutritional composition, palatability, physical form, and safety of their feed.

[0003] Although the pet food market has formed a diversified product system, including extruded feed for fish, insect feed, fruit puree feed, and paste feed for amphibians and reptiles, there are still many problems in terms of nutritional composition, feeding convenience, palatability, physical form, and safety.

[0004] Insect feed includes live insects, preserved insects, and freeze-dried insects. While live insects are highly palatable and can trigger natural predation behavior, they require suitable breeding space and experience, are prone to escape or death during breeding, pose a risk of carrying parasites, and have a short storage period (≤7 days). Preserved insects have a long shelf life, up to 12 months, and are portable, but added preservatives may affect pet health, and they require refrigeration at 4°C. Once opened, they are prone to mold, and their shelf life is ≤1 month. Freeze-dried insects retain some of their active odor, but freeze-drying is energy-intensive and costly. After rehydration, they become loose in texture, and the loss of volatile substances reduces their palatability. Fruit puree / paste-like feeds are convenient to feed and nutritionally balanced, but traditional sterilization processes, such as pasteurization, require high temperatures, which degrade heat-sensitive components and cannot completely kill heat-resistant spores. Furthermore, puree and paste-like foods provide no chewing feedback for pets, and their sticky surface can easily cause discomfort by sticking to the pet's mouth and nasal cavity. Additionally, their palatability relies on artificial flavorings, resulting in poor palatability. Extruded feeds, while stable in storage and low in cost, are hard and can easily damage the oral mucosa. After rehydration, they become loose and brittle, failing to mimic the hard-outer-soft-inner structure of insects. Moreover, the high temperatures destroy flavor compounds, further reducing their palatability.

[0005] Therefore, it is of great significance to develop a feed for poikilothermic animals that is highly palatable, retains a high percentage of nutrients, and is safe. Summary of the Invention

[0006] The present invention aims to solve one or more technical problems existing in the prior art, and at least provide a beneficial solution. Specifically, the present invention provides a poikilothermic animal feed, which has good palatability, can improve the feeding rate of pets, shorten the response time to aggression, and has a high nutrient retention rate and is safe.

[0007] The inventive concept of this invention: The cold-blooded animal feed of this invention has a core and a shell layer disposed on the surface of the core; the core includes sodium alginate, protein base, attractant A, and fermentation product synergist; the raw materials for preparing the shell layer include sodium alginate and calcium lactate; the shell layer includes attractant B; attractant A includes insect-derived attractant, animal-derived attractant, and plant-derived attractant; attractant B includes freeze-dried powder containing fructose and insect enzymatic hydrolysate.

[0008] Sodium alginate and calcium lactate undergo a cross-linking reaction to form a surface gel layer with a hardness of 1.5-2.0N. This essentially creates a deformation-resistant structure similar to an insect exoskeleton, effectively preventing food debris from entering the pet's respiratory tract during eating, significantly reducing the risk of suffocation and providing a solid guarantee for the pet's health. Furthermore, the hydrophilic migration properties of calcium lactate during gelation allow it to carry attractant B, which diffuses directionally to the gel surface, forming a "flavor layer" rich in attractant factors. This greatly enhances the dual stimulation of the pet's sense of smell and taste, significantly improving feeding induction efficiency. Simultaneously, the protein base, attractant A, and fermentation product synergist form a core structure with a hardness ≤0.3N. This unique "hard outside, soft inside" structure highly mimics the hard exoskeleton and soft interior of natural foods like insects, providing pets with a more natural hunting experience and increasing palatability.

[0009] Simultaneously, the natural flavor substances released from the endogenous base material, formed by the synergistic effect of insect-derived, animal-derived, and plant-derived attractants, and attractant B carried by the exogenous calcium source, create a gradient flavor field. This not only rapidly stimulates olfactory receptors on the surface but also prolongs taste stimulation through the continuous release of flavor substances from the core, overcoming the shortcomings of the single flavor release mode of traditional feeds, thus improving palatability and providing rich nutrition. Furthermore, the fermentation product synergist and the insect-derived attractant in attractant A can create a synergistic umami effect, enhancing the pet's feeding motivation and increasing the feeding rate, resulting in excellent palatability.

[0010] Therefore, a first aspect of the present invention provides a feed for poikilothermic animals.

[0011] Specifically, the poikilothermic animal feed has a core and a shell disposed on the surface of the core;

[0012] The core includes sodium alginate, protein base, palatability enhancer A, and fermentation product synergist.

[0013] The raw materials for preparing the shell include sodium alginate and calcium lactate; the shell contains a feeding attractant B.

[0014] The attractant A includes insect-derived attractants, animal-derived attractants, and plant-derived attractants;

[0015] The attractant B comprises a freeze-dried powder containing fructose and insect enzymatic hydrolysate.

[0016] Preferably, the insect-derived attractant includes at least one of cricket hydrolysate, mealworm hydrolysate, and insect oil extract.

[0017] Preferably, the insect oil extract includes at least one of mealworm oil, black soldier fly oil, and silkworm pupa oil.

[0018] Preferably, the animal-derived palatability enhancer includes at least one of chicken liver powder, hydrolyzed fish protein, and fish liver oil hydrolysate.

[0019] Preferably, the plant-derived palatability enhancer includes at least one of apple puree and carrot extract.

[0020] Specifically, insect-derived attractants provide the umami substance monosodium glutamate, animal-derived attractants provide the fishy substance trimethylamine, and plant-derived apple puree provides the fruity flavor substance ethyl acetate. The three form a synergistic flavor effect, which can shorten the pet's attack response time to food, increase the feeding rate, and thus improve palatability.

[0021] Preferably, in the attractant A, the mass ratio of the insect-derived attractant, animal-derived attractant, and plant-derived attractant is 1:(0.55-16.5):(0.35-4.5).

[0022] More preferably, in the attractant A, the mass ratio of the insect-derived attractant, animal-derived attractant, and plant-derived attractant is 1:(0.6-15):(0.4-4).

[0023] Preferably, the freeze-dried powder containing fructose includes at least one of honey freeze-dried powder and fruit freeze-dried powder.

[0024] Preferably, the fruit freeze-dried powder includes at least one of banana freeze-dried powder, strawberry freeze-dried powder, and blueberry freeze-dried powder.

[0025] Preferably, the insect enzymatic hydrolysate includes at least one of cricket enzymatic hydrolysate, mealworm enzymatic hydrolysate, black soldier fly enzymatic hydrolysate, and silkworm pupa enzymatic hydrolysate.

[0026] Specifically, the insect hydrolysate in attractant B contains ≥5% free amino acids, which, together with the honey fructose in the freeze-dried honey powder, form a sweet and fresh complex flavor. As it is a natural ingredient, the addition of natural attractants can greatly enhance the appeal of the feed to pets, stimulate their appetite, and improve the palatability of the feed.

[0027] Preferably, in the attractant B, the mass ratio of the fructose-containing freeze-dried powder to the insect enzymatic hydrolysate is 1:(0.6-1.5); more preferably, in the attractant B, the mass ratio of the fructose-containing freeze-dried powder to the insect enzymatic hydrolysate is 1:(0.8-1.2); even more preferably, in the attractant B, the mass ratio of the fructose-containing freeze-dried powder to the insect enzymatic hydrolysate is 1:1 or 1:1.5.

[0028] Preferably, the protein base material includes at least one of fish meal, shrimp meal, insect meal, blood meal, and spirulina powder.

[0029] Specifically, the protein-based feed provides pets with a high-quality protein source to meet their energy needs for growth and daily activities.

[0030] Preferably, the fermentation product enhancer includes at least one of yeast extract and lactic acid fermentation product.

[0031] Preferably, the core further includes plasticizers, additives, and solvents.

[0032] Preferably, the plasticizer includes at least one of glycerol and sorbitol.

[0033] Specifically, in the preparation of poikilothermic animal feed, plasticizers can be used to improve the flexibility and plasticity of the paste, enabling it to be smoothly shaped when extruded through different nozzles. Sorbitol also acts as a humectant to prevent the paste from hardening during storage.

[0034] Preferably, the additive includes at least one of vitamins and minerals; more preferably, the additive includes both vitamins and minerals to ensure that the pet receives comprehensive and balanced nutrition.

[0035] Preferably, the palatability enhancer B accounts for 0.45-2.2% of the total mass of the calcium lactate and palatability enhancer B; more preferably, the palatability enhancer B accounts for 0.5-2.0% of the total mass of the calcium lactate and palatability enhancer B.

[0036] A second aspect of the present invention provides a method for preparing poikilothermic animal feed as described in the first aspect of the present invention.

[0037] Specifically, the method for preparing the poikilothermic animal feed includes the following steps:

[0038] Sodium alginate, protein base, palatability enhancer A, fermentation product synergist, and solvent are mixed, filled, sterilized, and extruded to obtain a paste.

[0039] Calcium lactate, palatability enhancer B, and solvent are mixed to obtain a mixed solution;

[0040] The paste is immersed in the mixed solution and gelled to obtain the poikilothermic animal feed;

[0041] The sterilization process employs irradiation sterilization.

[0042] Preferably, the mixture is first degassed under vacuum to remove air mixed into the paste during stirring, so as to avoid affecting the quality of the paste due to the presence of air bubbles during subsequent processing or storage.

[0043] After degassing, the paste is filled into a tube. Select the appropriate nozzle size according to the pet's size and install it on the tube containing the paste. Due to the different nozzle sizes, the paste can be extruded into different shapes to meet the feeding needs of different pets.

[0044] Irradiation sterilization ensures feed safety while maximizing the retention of heat-sensitive nutrients, such as crude protein and crude fat, providing pets with superior nutritional support. Furthermore, irradiation sterilization offers greater reliability in microbial control, and the sterilized paste can be stored at room temperature, allowing for convenient long-term preservation and readily available use.

[0045] Preferably, by mass percentage, the amounts of each raw material used in the preparation of the paste are: sodium alginate 1.3-5.5%, protein base 22-65%, palatability enhancer A 0.9-4.5%, and fermentation product synergist 0.1-0.9%.

[0046] More preferably, by mass percentage, the amounts of each raw material used in the preparation of the paste are: sodium alginate 1.5-5%, protein base 25-60%, palatability enhancer A1-4%, and fermentation product synergist 0.1-0.8%.

[0047] Preferably, the preparation process of the paste further includes mixing with additives, plasticizers and solvents.

[0048] Preferably, by mass percentage, the amounts of each raw material used in the preparation of the paste are as follows: sodium alginate 1.3-5.5%, protein base 22-65%, palatability enhancer A 0.9-4.5%, fermentation product enhancer 0.1-0.9%, additives 0.1-3.3%, plasticizer 0.9-9%, and solvent 11.8-74.7%.

[0049] More preferably, by mass percentage, the amounts of each raw material used in the preparation of the paste are as follows: sodium alginate 1.5-5%, protein base 25-60%, palatability enhancer A 1-4%, fermentation product synergist 0.1-0.8%, additives 0.1-3%, plasticizer 1-8%, and solvent 19.2-71.3%.

[0050] Preferably, the solvent in the ointment includes water.

[0051] Preferably, in the preparation of the mixed solution, the ratio of the total mass of calcium lactate and palatability enhancer B to the amount of solvent is 1 g : (9-22) mL; more preferably, the ratio of the total mass of calcium lactate and palatability enhancer B to the amount of solvent is 1 g : (10-20) mL. A reasonable ratio ensures the smooth progress of the cross-linking reaction between calcium lactate and sodium alginate and the stability of the gel feed's quality.

[0052] Preferably, the solvent in the mixed solution includes water.

[0053] A third aspect of the present invention provides an application of the poikilothermic animal feed described in the first aspect of the present invention in the field of pet breeding.

[0054] Compared with the prior art, the beneficial effects of the technical solution provided by the present invention are as follows:

[0055] (1) In this invention, sodium alginate and calcium lactate undergo a cross-linking reaction to form a surface gel layer with a hardness of 1.5-2.0 N. Simultaneously, the protein base, palatability enhancer A, and fermentation product synergist form a core structure with a hardness ≤0.3 N. This unique "hard on the outside, soft on the inside" structure highly simulates the characteristics of the hard exoskeleton and soft interior of natural foods such as insects, providing pets with a palatability experience closer to natural predation. At the same time, the surface gel layer, similar to the deformation-resistant structure of an insect exoskeleton, can effectively prevent feed debris from entering the pet's respiratory tract during eating, greatly reducing the risk of suffocation and providing a solid guarantee for the pet's health.

[0056] (2) This invention achieves a significant improvement in feed feeding effectiveness through the design of a dual feeding system using attractant A and attractant B, coupled with attractant-calcium source coupling technology. Attractant B forms a "flavor layer" rich in attractant factors on the gel surface, greatly enhancing the dual stimulation of the pet's sense of smell and taste, and significantly improving feeding induction efficiency. The natural flavor substances released by the endogenous base material formed by the synergistic action of insect-derived, animal-derived, and plant-derived attractants, together with attractant B carried by the exogenous calcium source, form a gradient flavor field. This not only rapidly stimulates olfactory receptors on the surface but also prolongs taste stimulation through the continuous release of flavor substances from the core, overcoming the shortcomings of the traditional single flavor release mode of feed, improving feeding efficiency, and providing rich nutrition. In addition, the fermentation product synergist and the insect-derived attractant in attractant A can form a synergistic umami effect, enhancing the pet's feeding motivation and improving feeding efficiency, resulting in a good feeding effect.

[0057] (3) The combination of the low-temperature preparation process and the irradiation sterilization technology of this invention minimizes the damage to nutrients, strictly controls the protein denaturation rate to <5%, and the vitamin B2 retention rate to >95%, ensuring that pets can ingest comprehensive, balanced and highly active nutrients to meet their growth, development and daily activities.

[0058] (4) The feed sterilized by irradiation has good storage stability at room temperature and a shelf life of more than 12 months. At the same time, the tube packaging effectively isolates the intrusion of external air, moisture and microorganisms, further extending the shelf life of the product and reducing the risk of feed deterioration and nutrient loss.

[0059] (5) This invention is simple to operate, taking less than 10 minutes from extrusion of the paste to completion of gelation, requiring no complex equipment or professional skills. It is easy for both pet breeding companies and pet owners to use, meeting the needs of different scenarios, and is especially suitable for daily family feeding. Furthermore, different nozzle shapes (such as mouse-shaped, worm-shaped, etc.) can be selected according to the pet species to precisely control the size of the extruded paste, adapting to pets of different sizes. Simultaneously, by adjusting the concentration of the calcium lactate solution and the soaking time, the thickness of the gel layer can be autonomously controlled, and the size of the shaped paste and gel parameters can be adjusted to meet the needs of different growth stages. Attached Figure Description

[0060] Figure 1 This is a schematic diagram of the preparation process of poikilothermic animal feed in Example 1 of the present invention;

[0061] Figure 2 The graph shows the feed intake results of corn snakes on the poikilothermic animal feed of Example 1 and Comparative Examples 1-3;

[0062] Figure 3The graph shows the response time of corn snakes to attacks from poikilothermic animal feed in Example 1 and Comparative Examples 1-3.

[0063] Figure 4 Figure showing the effect of different sterilization methods on crude protein and crude fat;

[0064] Figure 5 The graph shows the effects of different sterilization methods on vitamin B2 and vitamin C. Detailed Implementation

[0065] To enable those skilled in the art to more clearly understand the technical solutions described in this invention, the following embodiments are provided for illustration. It should be noted that the following embodiments do not constitute a limitation on the scope of protection claimed by this invention.

[0066] Unless otherwise specified, the raw materials, reagents or devices used in the following examples are available from conventional commercial sources or can be obtained by existing known methods.

[0067] In Examples 1-3, the vitamin premix consists of the following substances: Vitamin A: 25000 IU / kg, D-Biotin: 0.3 mg / kg, D-Calcium Pantothenate: 30 mg / kg, Choline Chloride: 2000 mg / kg, Vitamin D3: 5500 IU / kg, Folic Acid: 3 mg / kg, α-Tocopherol: 330 mg / kg, Vitamin K3: 3 mg / kg, Vitamin B1: 30 mg / kg, Vitamin B2: 30 mg / kg, Vitamin B6: 30 mg / kg, Vitamin B12: 0.055 mg / kg, Niacin: 60 mg / kg, and Inositol: 300 mg / kg.

[0068] In Examples 1-3, the mineral premix consists of the following substances: glycine iron complex: 125 mg / kg, amino acid zinc complex: 100 mg / kg, methionine manganese complex: 60 mg / kg, glycine copper complex: 30 mg / kg, calcium iodate: 1.1 mg / kg, cobalt chloride: 0.55 mg / kg, sodium selenite: 0.3 mg / kg, magnesium sulfate: 600 mg / kg, and potassium chloride: 1250 mg / kg.

[0069] Example 1

[0070] A method for preparing cold-blooded animal feed for pet snakes includes the following steps:

[0071] (1) Preparation of paste

[0072] S1: Weigh 0.35 kg of sodium alginate and 0.5 kg of sorbitol and dry mix them. Add 4.65 L of purified water at 45 °C and stir magnetically at 300 rpm for 1 hour until completely dissolved. The solution should be transparent and free of visible particles. Then add 2.5 kg of spray-dried pig blood powder, 1.4 kg of fish meal, 0.06 kg of vitamin premix, 0.06 kg of mineral premix, 0.2 kg of chicken liver powder, 0.1 kg of hydrolyzed fish protein, 0.08 kg of apple puree, 0.02 kg of cricket enzymatic hydrolysate, 0.05 kg of yeast extract, and 0.03 kg of lactic acid fermentation product. Transfer the mixture to a high-speed homogenizer and homogenize for 15 minutes to obtain a mixed paste.

[0073] S2: Transfer the mixed paste to a vacuum degassing tank for vacuum degassing at a vacuum degree of -0.09MPa for 35 minutes until no bubbles are released; fill the tubes with a diameter of 30mm and a capacity of 200mL, then heat seal them at a temperature of 180℃, a pressure of 0.3MPa, and a time of 2 seconds. Finally, sterilize the paste by cobalt-60 gamma irradiation at a dose of 4.0kGy, a dose uniformity (DUR) of ≤1.4, and an ambient temperature of 25±2℃. After irradiation, cool the paste to room temperature and store it away from light to obtain the radiation-sterilized paste.

[0074] (2) Preparation of calcium lactate mixed solution

[0075] Weigh out 9.8 kg of calcium lactate, 0.1 kg of freeze-dried honey powder, and 0.1 kg of black soldier fly hydrolysate. First, transfer the freeze-dried honey powder and black soldier fly hydrolysate to a mixer and mix at 30 rpm for 15 minutes. Then add calcium lactate and continue mixing for 20 minutes. Pass the mixture through an 80-mesh sieve to obtain a mixed powder. Take 10 g of the mixed powder, add 200 mL of purified water at 25°C, and stir for about 2 minutes until transparent. The pH is 6.8 ± 0.2, thus obtaining a calcium lactate mixed solution.

[0076] (3) Surface gelation

[0077] Select a cylindrical nozzle with a diameter of 10mm × 20mm (10mm in diameter and 20mm in length) to simulate the body shape of snake prey (such as pinkie mice). Tighten the nozzle to the opening of the hose and vertically squeeze the radiation-sterilized paste from the hose into the 5% calcium lactate mixed solution obtained in step (2). Let it stand for 4 minutes. The paste will gel in the calcium lactate mixed solution and be freely shaped into a cold-temperature animal feed for pet snakes that mimics pinkie mice. It can be directly picked up and fed.

[0078] Example 1: Schematic diagram of the preparation process of poikilothermic animal feed. Figure 1 As shown.

[0079] Example 2

[0080] A method for preparing poikilothermic animal feed for geckos includes the following steps:

[0081] (1) Preparation of paste

[0082] S1: Weigh out 0.3 kg of sodium alginate and 0.4 kg of glycerin, mix them dry, add 4.7 L of purified water at 45 °C, and stir magnetically at 300 rpm for 1 hour until completely dissolved. The solution should be transparent and free of visible particles. Then add 3 kg of cricket powder, 1.2 kg of mealworm powder, 0.08 kg of vitamin premix, 0.08 kg of mineral premix, 0.05 kg of chicken liver powder, 0.03 kg of apple puree, 0.08 kg of black soldier fly enzymatic hydrolysate, 0.03 kg of yeast extract, and 0.05 kg of lactic acid fermentation product. Transfer the mixture to a high-speed homogenizer and homogenize for 15 minutes to obtain a mixed paste.

[0083] S2: Transfer the mixed paste obtained in step S1 to a vacuum degassing tank for vacuum degassing at a vacuum degree of -0.08 MPa for 30 minutes until no bubbles are released; then fill it into a 30 mm diameter, 200 mL tubing, heat seal it at a temperature of 180°C, a pressure of 0.3 MPa, and a time of 2 seconds; finally, sterilize it by cobalt-60 gamma irradiation at a dose of 10 kGy, a dose uniformity (DUR) ≤1.5, and an ambient temperature of 25 ± 2°C. After irradiation, cool it to room temperature and store it in the dark to obtain the radiation-sterilized paste.

[0084] (2) Preparation of calcium lactate mixed solution

[0085] Weigh out 9.9 kg of calcium lactate, 0.05 kg of freeze-dried honey powder, and 0.05 kg of cricket enzymatic hydrolysate. First, transfer the freeze-dried honey powder and black soldier fly enzymatic hydrolysate to a mixer and mix at 30 rpm for 15 minutes. Then add calcium lactate and continue mixing for 20 minutes. Pass the mixture through an 80-mesh sieve to obtain a mixed powder. Take 15 g of the mixed powder, add 300 mL of purified water at 25°C, and stir for about 2 minutes until transparent. The pH is 6.8 ± 0.2, thus obtaining a calcium lactate mixed solution.

[0086] (3) Surface gelation

[0087] Select a cylindrical nozzle with a diameter of 5mm × 10mm (5mm in diameter and 10mm in length) and a strip-shaped nozzle that mimics the shape of a worm. Tighten the nozzle to the opening of the hose and vertically squeeze the radiation-sterilized paste from the hose into the 5% calcium lactate mixed solution obtained in step (2). Let it stand for 3 minutes. The paste will gel in the calcium lactate mixed solution and be freely shaped into a cold-blooded animal feed for geckos that mimics worms. It can be directly picked up and fed.

[0088] Example 3

[0089] A method for preparing poikilothermic animal feed for horned frogs includes the following steps:

[0090] (1) Preparation of paste

[0091] S1: Weigh 0.4 kg of sodium alginate and 0.5 kg of glycerin and dry mix them. Add 4.1 L of purified water at 45°C and stir magnetically at 600 rpm for 1 hour until completely dissolved and no visible particles remain. Then add 3.6 kg of fish meal, 1 kg of Antarctic krill powder, 0.05 kg of spirulina powder, 0.05 kg of vitamin premix, 0.05 kg of mineral premix, 0.1 kg of cod liver oil hydrolysate, 0.05 kg of hydrolyzed fish protein, 0.05 kg of carrot extract, 0.03 kg of mealworm hydrolysate, 0.01 kg of yeast extract, and 0.01 kg of lactic acid fermentation product. Transfer the mixture to a high-speed homogenizer and homogenize for 15 minutes to obtain a mixed paste.

[0092] S2: Transfer the mixed paste to a vacuum degassing tank for vacuum degassing at a vacuum degree of -0.09 MPa for 35 minutes until no bubbles are released. Then, fill the tubes with a diameter of 30 mm and a capacity of 200 mL, heat-seal them at a temperature of 180℃, a pressure of 0.3 MPa, and a time of 2 seconds. Finally, sterilize the paste by cobalt-60 gamma irradiation at a dose of 3.5 kGy, a dose uniformity (DUR) ≤ 1.4, and an ambient temperature of 25 ± 2℃. After irradiation, cool the paste to room temperature and store it away from light to obtain the radiation-sterilized paste.

[0093] (2) Preparation of calcium lactate mixed solution

[0094] Weigh out 9.95 kg of calcium lactate, 0.02 kg of freeze-dried honey powder, and 0.03 kg of mealworm enzymatic hydrolysate. First, transfer the freeze-dried honey powder and mealworm enzymatic hydrolysate to a mixer and mix at 30 rpm for 15 minutes. Then add calcium lactate and continue mixing for 20 minutes. Pass the mixture through an 80-mesh sieve to obtain a mixed powder. Take 20 g of the mixed powder and add it to 300 mL of purified water at 25°C. Stir for about 2 minutes until transparent, with a pH of 7.0 ± 0.2, to obtain a calcium lactate mixed solution.

[0095] (3) Surface gelation operation

[0096] Select a cylindrical nozzle with a diameter of 5mm × 15mm (5mm in diameter and 15mm in length) to match the oral cavity size of horned frogs with a body length of 8-12cm. Tighten the nozzle to the opening of the hose and vertically squeeze the radiation-sterilized paste from the hose into the 6% calcium lactate solution obtained in step (2). Let it stand for 5 minutes. The paste will gel in the calcium lactate solution and be freely shaped into cold-blooded animal feed for horned frogs, which can be directly picked up and fed.

[0097] Comparative Example 1

[0098] The only difference between Comparative Example 1 and Example 1 is that Comparative Example 1 uses an animal-derived attractant to replace the insect-derived attractant in an equal amount, that is, 0.02 kg of chicken liver powder is used to replace the cricket enzymatic hydrolysate in an equal amount. Otherwise, it is the same as Example 1.

[0099] Comparative Example 2

[0100] The only difference between Comparative Example 2 and Example 1 is that Comparative Example 2 uses an insect-derived attractant to replace the animal-derived attractant in an equal amount, that is, 0.3 kg of cricket enzymatic hydrolysate is used to replace the animal-derived attractant in an equal amount. Otherwise, it is the same as Example 1.

[0101] Comparative Example 3

[0102] The only difference between Comparative Example 3 and Example 1 is that Comparative Example 3 uses an animal-derived attractant to replace the plant-derived attractant in an equal amount, that is, 0.08 kg of chicken liver powder is used to replace apple puree in an equal amount. Otherwise, it is the same as Example 1.

[0103] Comparative Example 4

[0104] The only difference between Comparative Example 4 and Example 2 is that Comparative Example 4 uses 0.08 kg of maltose to replace the fermentation product enhancer (0.03 kg of yeast extract and 0.05 kg of lactic acid fermentation product), i.e., it does not contain the fermentation product enhancer. Otherwise, it is the same as Example 2.

[0105] Comparative Example 5

[0106] The only difference between Comparative Example 5 and Example 1 is that Comparative Example 5 uses pasteurization, which involves heating the filled tubing in a water bath at 63°C for 30 minutes. The rest is the same as in Example 1.

[0107] Performance testing

[0108] 1. Feed intake rate test

[0109] The feeding rate of corn snakes on the poikilothermic animal feed prepared in Example 1 and Comparative Examples 1-3 was tested, and the specific process is as follows:

[0110] Forty corn snakes (Pantherophis guttatus) of the same species, age, good health, and similar size were selected and randomly divided into four groups of 10 snakes each. Each group of snakes was placed in an independent enclosure, with the ambient temperature controlled at 28-30℃ and the humidity maintained at 60-70%. Suitable hiding places and clean drinking water were provided.

[0111] Before the experiment, the snakes were allowed to acclimatize to their respective enclosures for 7 days, during which they were fed once a day to ensure they adapted to the environment and had a normal appetite. At the start of the experiment, they were fed once every 5 days, for a total of 10 feedings. Each feeding was scheduled for 10:00 AM. Appropriate poikilothermic animal feed (Example 1, Comparative Example 1, Comparative Example 2, Comparative Example 3) was placed in each enclosure. The amount of feed was determined based on the snake's size, approximately 10% of its body weight per snake. After feeding, the snakes were observed for 30 minutes, and whether they ate was recorded. If a snake bit and swallowed the feed, it was considered to have eaten; if it only smelled or touched the feed but did not swallow, it was considered not to have eaten. The feeding rate was then calculated using the following formula:

[0112] Feeding rate = (Total number of feedings per group / (Number of snakes per group × Number of experimental days)) × 100%.

[0113] The feed intake results of corn snakes on poikilothermic animal diets in Examples 1 and 1-3 are as follows: Figure 2 As shown.

[0114] Depend on Figure 2 It can be seen that Example 1 achieved the highest feeding rate, with an average of 96%, through the synergistic effect of insect, animal, and plant-derived attractants. Comparative Example 1, where an equal amount of animal-derived attractant replaced the insect-derived attractant, resulted in a surge in fishy-smelling substances leading to a rejection reflex, and the feeding rate dropped to 86%, with significant fluctuations. Comparative Example 2, where an equal amount of insect-derived attractant replaced the animal-derived attractant, improved the insect umami flavor, but lacked a fishy-smelling buffer, resulting in an average feeding rate of 88% and taste fatigue. Comparative Example 3, where an equal amount of animal-derived attractant replaced the plant-derived attractant, completely lost the fruity aroma modulation, resulting in the lowest feeding rate of 81%. Analysis of variance showed significant differences among the groups (p<0.05), indicating that the ternary system of insect-derived attractants providing core umami, animal-derived attractants triggering predatory instincts, and plant-derived attractants regulating flavor balance is irreplaceable. The combined effect of all three effectively improves the feeding rate, and the absence of any one component leads to a significant decrease in the feeding rate.

[0115] 2. Measurement of attack response time

[0116] The test examined the attack response time of corn snakes to the poikilothermic animal feed prepared in Example 1 and Comparative Examples 1-3. The specific process is as follows:

[0117] A high-definition camera was placed in a location where the snakes and feed could be clearly seen to record the entire experimental process. The grouping of corn snakes and the use of cold-blooded animal feed were the same as those used in the above-mentioned feeding rate determination. Timing was started from the moment the feed entered the enclosure. The time of the first attack action (such as suddenly raising its head or opening its mouth to bite the feed) of the corn snake was recorded by video playback, accurate to 0.1 seconds. If the snake did not make an attack action within 30 minutes, it was recorded as no response.

[0118] Attack response time is calculated using the following formula:

[0119] Attack response time = (sum of attack response times of each corn snake in the group) / number of corn snakes in each group.

[0120] The response time of corn snakes to attacks from poikilothermic animal diets in Examples 1 and 1-3 is as follows: Figure 3 As shown.

[0121] Depend on Figure 3 It was found that Example 1 had the shortest attack response time, with an average of 1.54 seconds. This indicates that, under the synergistic effect of insect, animal, and plant-derived attractants, poikilothermic animal feed has a strong attraction for corn snakes and can quickly stimulate their attack behavior. This may be because this synergistic effect makes the feed more similar in flavor and texture to the characteristics of the corn snake's natural prey, thereby triggering the corn snake's predatory instincts and enabling it to make an attack response in a shorter time.

[0122] Comparative Example 1, where an equal amount of animal-derived attractant was used to replace the insect-derived attractant, showed an average attack response time of 3.27 seconds, significantly longer than Example 1. This may be because replacing the insect-derived attractant with an animal-derived attractant alters the flavor of the feed, potentially increasing the amount of fishy substances, which reduces the corn snake's recognition and acceptance of the feed, requiring more time to react or making a more cautious attack decision.

[0123] Comparative Example 2, which replaced the animal-derived attractant with an equal amount of insect-derived attractant, showed an average attack response time of 2.58 seconds, significantly higher than Example 1. Although the increase in insect-derived attractant enhanced certain flavor components, the reduction in animal-derived components may have altered the overall flavor and texture balance of the feed, failing to achieve the same close resemblance to natural prey as in Example 1, thus prolonging the attack response time of corn snakes.

[0124] Comparative Example 3, where an animal-derived attractant was used to replace the plant-derived attractant in equal amounts, resulted in the longest attack response time, averaging 4.27 seconds. This may be because the lack of plant-derived attractant made the feed flavor monotonous, failing to effectively stimulate the corn snake's olfactory and gustatory receptors, thus reducing its interest in the feed. Consequently, the corn snake required a longer time to respond to an attack, and may even become confused or unaccustomed to the feed.

[0125] In summary, insect, animal, and plant-derived attractants have a synergistic effect. A complete ternary attractant system of insect, animal, and plant-derived attractants can significantly shorten the attack response time. The absence of any one type of attractant will lead to a prolonged attack response time, with the lack of plant-derived attractants having the most significant impact.

[0126] That is, the cold-blooded animal feed of Embodiment 1 of the present invention can improve the feeding rate of corn snakes, shorten the attack response time, and has good palatability.

[0127] 3. Synergistic effect test of umami flavor between fermentation product enhancers and insect enzymatic hydrolysates

[0128] The following experimental procedure was used to verify the synergistic effect of fermentation product enhancers and insect enzymatic hydrolysates on umami flavor. The specific procedure is as follows:

[0129] Twenty leopard geckos (Eublepharismacularius) of the same species, age, good health and similar size were selected and randomly divided into two groups of 10 geckos each, weighing 30-40g. Each gecko was placed in an independent enclosure with an ambient temperature of 28±1℃, humidity of 60±5%, and a light cycle of 12 hours of light and 12 hours of darkness. Suitable hiding places and clean drinking water were also provided.

[0130] Before the experiment, the geckos were allowed to acclimatize in their respective enclosures for 7 days, during which they were fed normally. The lights were turned off during feeding to simulate the dark environment of the experiment, ensuring that the geckos adapted to the environment and had a normal appetite. At the beginning of the experiment, they were fed once every 3 days (for a total of 3 cycles). Fresh feed (the poikilothermic animal feed of Example 2 and Comparative Example 4) was given at 18:00 every evening, and the amount of feed was 10% of their body weight (accurate to 0.1g). The feeding amount was adjusted weekly by weighing. The feed was fixed in the center of the enclosure during feeding, and the enclosure was cleaned after feeding to avoid residual interference. The lights were turned off during the experiment, and only a weak red light was used for observation.

[0131] Record indicators: first feeding time (the number of seconds from when the feed is placed to when the gecko first touches it), calculate the feeding rate (%), and the method for calculating the feeding rate is the same as above.

[0132] In addition, the contents of umami substances, such as free monosodium glutamate (Glu), disodium 5'-inosinate (IMP), disodium 5'-guanylate (GMP), and total free amino acids (FAA), in the poikilothermic animal feeds of Example 2 and Comparative Example 4 were measured. Simultaneously, a sensory evaluation team composed of 10 trained sensory evaluators conducted a sensory evaluation of the umami intensity, scoring it from 1 to 10.

[0133] Free monosodium glutamate was tested according to GB 7300.1001-2020; disodium 5'-inosinate and disodium 5'-guanylate were tested according to T / SDFA 038-2023; and total free amino acids were tested according to GB / T 18246-2019.

[0134] The results of the umami content and umami intensity of the poikilothermic animal feed in Example 2 and Comparative Example 4, as well as the first feeding time and feeding rate of geckos, are shown in Table 1.

[0135] Table 1: Content and intensity of umami substances in poikilothermic animal feeds of Example 2 and Comparative Example 4; time of first feeding and feeding rate of geckos.

[0136] index Example 2 Comparative Example 4 p-value Total Glu (g / kg) 4.10±0.82 4.03±0.75 ＞0.05 IMP + GMP (g / kg) 0.032±0.007 0.000±0.000 ＜0.01 Total FAA (g / kg) 7.25±0.95 7.12±0.90 ＞0.05 Umami intensity rating / point 8.5±0.5 6.2±0.8 ＜0.01 First feeding time / s 28±5 55±10 ＜0.01 Food intake rate / % 98.6±1.2 82.1±3.5 ＜0.01

[0137] As can be seen from Table 1, the cold-blooded animal feed of Example 2 of the present invention contains umami substances IMP and GMP, and the basic umami substances monosodium glutamate (Glu) and total free amino acids (FAA) are slightly higher, resulting in a significant umami intensity. This can attract pets to eat, reduce the time to first feeding, and increase the feeding rate.

[0138] The initial feeding time in Example 2 was significantly shorter than that in Comparative Example 4, and the feeding rate was significantly increased. This indicates that the synergistic effect of the fermentation product enhancer and the umami of the insect enzymatic hydrolysate directly enhanced the gecko's feeding motivation. Immediate appeal stems from the high concentration of umami substances in the surface flavor enrichment layer, while sustained palatability benefits from the gradient stimulation formed by the slow release of Glu and nucleotides from the core matrix.

[0139] In Comparative Example 4, the total monosodium glutamate (Glu) and total free amino acid (FAA) content were not significantly different from those in Example 2 (p>0.05), indicating that the absence of the fermentation product enhancer did not affect the composition of these basic umami substances in the feed. However, due to the lack of the fermentation product enhancer, Comparative Example 4 lacked the umami substances IMP and GMP, and the umami intensity was significantly reduced. The synergistic coefficient (Q value) calculation (Q is approximately 1.27) confirmed a nonlinear synergistic effect between nucleotides and Glu (Q>1). This may be because, from a molecular mechanism perspective, the heterodimeric structure of the umami receptor T1R1 / T1R3 has a synergistic response to the binding of nucleotides and Glu. Glu activates the receptor, while nucleotides prolong the receptor activation time, increase the receptor's affinity for Glu, and enhance umami perception.

[0140] The Q value is a quantitative indicator that measures whether the combined action of two or more substances produces a synergistic effect. Here, it is used to measure the synergistic effect when monosodium glutamate (Glu) and nucleotides (IMP+GMP) are combined.

[0141] The calculation formula is: Q = Actual observed effect (umami intensity of combined effect) / Theoretical expected effect (sum of individual effects). If Q = 1, it means that the combined effect equals the sum of individual effects (no synergistic effect); if Q > 1, it means that the combined effect exceeds the sum of individual effects (synergistic effect exists).

[0142] The calculation process for the Q value in this experiment is as follows:

[0143] Actual observation results: The umami intensity score of Example 2 was 8.5 points, representing the actual effect of the combined action of Glu and (IMP+GMP);

[0144] Theoretical expected effect: The umami intensity score of Comparative Example 4 is 6.2 points + theoretical contribution of 0.5 points from the effect of nucleotides alone.

[0145] Q = 8.5 / (6.2+0.5)≈1.27>1, indicating that the combined effect of Glu and (IMP+GMP) is not a simple additive effect, but rather a synergistic effect produced through molecular mechanisms.

[0146] The above comparison shows that fermentation product enhancers and insect enzymatic hydrolysates have a synergistic effect on umami flavor. Fermentation product enhancers are irreplaceable. The combined effect of the two can enrich umami substances, increase the umami intensity of poikilothermic animal feed, thereby reducing the time to first feeding and increasing the feeding rate.

[0147] 4. The effect of different sterilization methods on the retention rate of nutrients and sterilization effect of ointments

[0148] The retention rate of nutrients and the content of microorganisms of poikilothermic animal feed prepared by different sterilization methods in Example 1 and Comparative Example 5 were tested. Poikilothermic animal feed prepared without sterilization during preparation (i.e., compared with Example 1, the only difference is that the paste is kept sealed and not sterilized) was used as the control group.

[0149] The nutritional components tested included crude protein, crude fat, vitamin B2, and vitamin C, which were determined using the Kjeldahl method, Soxhlet extraction method, high performance liquid chromatography, and 2,6-dichlorophenolindophenol titration method, respectively.

[0150] The types of microorganisms involved in the test include bacteria, Salmonella, coliforms, and molds. The total bacterial count was determined using the plate count method, Salmonella content was determined according to GB / T 13091-2018, and coliforms and mold content were determined according to GB / T 18869-2019 and GB / T13092-2006, respectively.

[0151] The effects of different sterilization methods on crude protein and crude fat are as follows: Figure 4 As shown, the effects of different sterilization methods on vitamin B2 and vitamin C are as follows: Figure 5 As shown.

[0152] Depend on Figure 4It can be seen that the crude protein and crude fat contents in the poikilothermic animal feed of the irradiation sterilization group in Example 1 were 47.2 g / 100 g and 17.7 g / 100 g, respectively, which were very close to those of the control group, with a retention rate of 98.3% for both. In contrast, the crude protein and crude fat contents in Comparative Example 5 were 44.6 g / 100 g and 16.8 g / 100 g, respectively, with retention rates of 92.9% and 93.3%. The crude protein and crude fat retention rates of Example 1 were higher than those of Comparative Example 5 (crude protein loss of 7.1% and crude fat loss of 6.7%).

[0153] Depend on Figure 5 It can be seen that, for heat-sensitive vitamins, the vitamin B2 and vitamin C contents in the poikilothermic animal feed of the irradiation sterilization group in Example 1 were 0.81 mg / 100g and 24.2 mg / 100g, respectively, which are close to those of the control group. The retention rates of vitamin B2 and vitamin C in Example 1 were 95.3% and 96.8%, respectively. In contrast, the vitamin B2 and vitamin C contents in the poikilothermic animal feed of the pasteurization group in Comparative Example 5 were 0.62 mg / 100g and 18.5 mg / 100g, respectively, with nutrient retention rates of 72.9% and 74.0%, respectively.

[0154] That is, the nutrient retention rate of the poikilothermic animal feed prepared by radiation sterilization in Example 1 was significantly higher than that in Comparative Example 5. This difference stems from the non-thermal effect of irradiation sterilization, which directly destroys microbial DNA through gamma rays, avoiding the damage to protein spatial structure and vitamin activity caused by high-temperature treatment.

[0155] The sterilization effects of different sterilization methods are shown in Table 2.

[0156] Table 2: Sterilization effects of different sterilization methods

[0157]

[0158] As shown in Table 2, the total bacterial count, coliform count, and mold count in the irradiation sterilization group were significantly lower than those in the pasteurization group, and all fully met the standard requirements. This indicates that irradiation sterilization has higher reliability in terms of microbial control.

[0159] In other words, Embodiment 1 of the present invention uses irradiation sterilization technology to ensure feed safety while preserving heat-sensitive nutrients to the greatest extent, providing pets with better nutritional protection.

[0160] In addition, the poikilothermic animal feeds prepared in Examples 2 and 3 of this invention also have good palatability, umami intensity, nutrient retention rate, and bactericidal effect.

[0161] In summary, this invention improves feeding rates and shortens attack response time through the synergistic effect of insect-derived, animal-derived, and plant-derived attractants, demonstrating excellent palatability. The synergistic effect of fermentation product synergists and insect enzymatic hydrolysates creates a savory synergistic effect, enhancing pets' feeding motivation and improving feeding efficiency, resulting in excellent palatability. Furthermore, the combination of low-temperature preparation and irradiation sterilization technology improves nutrient retention and provides excellent sterilization, ensuring pets receive comprehensive, balanced, and highly active nutrients to meet their growth, development, and daily activity needs.

[0162] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A feed for poikilothermic animals, characterized in that, The poikilothermic animal feed has a core and a shell disposed on the surface of the core; The core includes sodium alginate, protein base, palatability enhancer A, and fermentation product synergist. The raw materials for preparing the shell include sodium alginate and calcium lactate; the shell contains a feeding attractant B. The attractant A includes insect-derived attractants, animal-derived attractants, and plant-derived attractants; The attractant B comprises a freeze-dried powder containing fructose and insect enzymatic hydrolysate; The insect-derived attractant includes at least one of cricket enzymatic hydrolysate, mealworm enzymatic hydrolysate, and insect oil extract; the animal-derived attractant includes at least one of chicken liver powder, hydrolyzed fish protein, and cod liver oil enzymatic hydrolysate. The plant-derived palatability enhancer includes at least one of apple puree and carrot extract; The freeze-dried powder containing fructose includes at least one of honey freeze-dried powder and fruit freeze-dried powder; The insect enzymatic hydrolysate includes at least one of cricket enzymatic hydrolysate, yellow mealworm enzymatic hydrolysate, black soldier fly enzymatic hydrolysate, and silkworm pupa enzymatic hydrolysate; The fermentation product enhancer includes at least one of yeast extract and lactic acid fermentation product. In attractant A, the mass ratio of the insect-derived attractant, animal-derived attractant, and plant-derived attractant is 1:(0.55-16.5):(0.35-4.5); in attractant B, the mass ratio of the freeze-dried powder containing fructose to the insect enzymatic hydrolysate is 1:(0.6-1.5).

2. The poikilothermic animal feed according to claim 1, characterized in that, The protein base material includes at least one of fish meal, shrimp meal, insect meal, blood meal, and spirulina powder.

3. The poikilothermic animal feed according to claim 1, characterized in that, The core also includes additives, plasticizers and solvents; the palatability enhancer B accounts for 0.45-2.2% of the total mass of the calcium lactate and palatability enhancer B.

4. The method for preparing poikilothermic animal feed according to any one of claims 1-3, characterized in that, The preparation method includes the following steps: Sodium alginate, protein base, palatability enhancer A, and fermentation product synergist are mixed, filled, sterilized, and extruded to obtain a paste. Calcium lactate, palatability enhancer B, and solvent are mixed to obtain a mixed solution; The paste is immersed in the mixed solution and gelled to obtain the poikilothermic animal feed; The sterilization process employs irradiation sterilization.

5. The preparation method according to claim 4, characterized in that, By mass percentage, the amounts of each raw material used in the preparation of the paste are as follows: sodium alginate 1.3-5.5%, protein base 22-65%, palatability enhancer A 0.9-4.5%, and fermentation product synergist 0.1-0.9%; In the preparation of the mixed solution, the ratio of the total mass of the calcium lactate and the palatability enhancer B to the amount of the solvent is 1 g: (9-22) mL.

6. The preparation method according to claim 4, characterized in that, The preparation process of the paste also includes mixing with additives, plasticizers and solvents.

7. The application of the poikilothermic animal feed according to any one of claims 1-3 in the field of pet breeding.

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