Pearl gentian stone fish feed

Abstract

The application provides a pearl gynura Gynura japonica feed and a preparation method thereof, and comprises the following components: 0.01-0.07% of methionine chromium, 26-27.5% of fish meal, 14-16% of soybean meal, 9-10% of soybean protein concentrate, 18-19.5% of chicken meal, 15.45-20.45% of cassava starch, 0.1-0.12% of methionine, 4-4.5% of fish oil, 0.9-1.1% of vitamin premix, 0.4-0.6% of mineral premix, 1.2-1.3% of sodium carboxymethyl cellulose, 1.4-1.6% of betaine, 0.004-0.006% of spermidine, 0.25-0.28% of isoleucine and 0.6-6.0% of defatted bone meal. The application can not only promote the protein digestion and absorption capacity of the pearl gynura Gynura japonica, enhance the antioxidant defense level, reduce oxidative stress and improve the liver function, but also promote the improvement of the health state of the body, so as to provide strong nutritional support for the realization of the rapid and healthy growth of the fish body and the formation of excellent meat quality.

Description

Technical Field

[0001] This invention relates to the field of animal husbandry technology, and more specifically, to a feed for pearl grouper. Background Technology

[0002] Chromium methionine is an organic compound formed by the combination of chromium and methionine. It is primarily used to regulate blood sugar, improve insulin sensitivity, and promote metabolic health. In nutrition, it is considered a trace element supplement that helps maintain glucose and lipid metabolism balance by enhancing the bioavailability of chromium. Chromium methionine participates in insulin signaling, helping cells absorb glucose more efficiently. Methionine, as an amino acid carrier, can enhance the absorption rate of chromium, which participates in protein synthesis and fat breakdown. Methionine itself is an important raw material for protein synthesis; the combination of chromium and chromium may synergistically promote muscle repair and reduce body fat accumulation. Methionine contains sulfur groups, which have antioxidant properties and can reduce free radical damage.

[0003] In recent years, with the rapid development of the global aquaculture industry, especially the increasing demand in the high-end aquatic product market, the pearl grouper, as a high-quality marine aquaculture species, has seen its farming scale and output continuously expand. However, how to improve farming efficiency, shorten the farming cycle, and simultaneously ensure the health and meat quality of the fish has become a key issue that the industry urgently needs to address. Existing feed formulations generally suffer from problems such as unbalanced nutrient ratios and insufficient functional components, making it difficult to meet the nutritional needs of the pearl grouper's rapid growth. Therefore, developing a new feed formulation that can significantly promote the growth of the pearl grouper not only has significant scientific research value but also broad market application prospects. Summary of the Invention

[0004] In view of this, the present invention proposes a pearl giant grouper feed, which aims to solve the problems of unbalanced nutritional ratio and insufficient functional components in existing pearl giant grouper feeds, making it difficult to effectively support their rapid and healthy growth and ensure meat quality. This invention proposes a feed for pearl grouper, comprising, by weight percentage: 0.01-0.07% chromium methionine, 26-27.5% fish meal, 14-16% soybean meal, 9-10% soybean protein concentrate, 18-19.5% chicken meal, 15.45-20.45% tapioca starch, 0.1-0.12% methionine, 4-4.5% fish oil, 0.9-1.1% vitamin premix, 0.4-0.6% mineral premix, 1.2-1.3% sodium carboxymethyl cellulose, 1.4-1.6% betaine, 0.004-0.006% spermidine, 0.25-0.28% isoleucine, and 0.6-6.0% degummed bone meal.

[0005] Furthermore, the above-mentioned pearl grouper feed, by weight percentage, includes: 0.03-0.07% chromium methionine, 26.78% fish meal, 15% soybean meal, 9.45% soybean protein concentrate, 18.78% chicken meal, 15.45-20.45% tapioca starch, 0.11% methionine, 4.2% fish oil, 1% vitamin premix, 0.5% mineral premix, 1.25% sodium carboxymethyl cellulose, 1.5% betaine, 0.005% spermidine, 0.26% isoleucine, and 0.665-5.715% degummed bone meal.

[0006] Furthermore, in the above-mentioned pearl grouper feed, the chromium methionine is added to the basic feed at a ratio of 0.3~0.7 g / kg, and the cassava starch content of the basic feed is 15.45%.

[0007] Furthermore, in the above-mentioned pearl grouper feed, the chromium methionine is added to the basal feed at a ratio of 0.5 to 0.7 g / kg.

[0008] Furthermore, in the above-mentioned pearl grouper feed, the chromium methionine is added to the high-sugar basal feed at a ratio of 0.5 g / kg, wherein the cassava starch content of the high-sugar basal feed is 20.45%.

[0009] Furthermore, in the aforementioned pearl grouper feed, the vitamin premix comprises, per kilogram of premix, the following components: vitamin B1 2.5 g / kg, vitamin B2 10 g / kg, vitamin B6 2.5 g / kg, vitamin B... 12 0.05g / kg, Vitamin C 14.84g / kg, Vitamin D3 0.0028g / kg, Vitamin E 6.3g / kg, Vitamin K 2g / kg, Niacin 37.5g / kg, Biotin 0.25g / kg, Calcium Pantothenate 25g / kg, Folic Acid 0.75g / kg, Inositol 26.79g / kg, Vitamin A 1g / kg, Fiber 870.5172g / kg.

[0010] Furthermore, in the above-mentioned pearl grouper feed, the mineral premix, per kilogram of premix, comprises the following components: calcium lactate 327 g / kg, potassium chloride 205.28 g / kg, calcium chloride 87.58 g / kg, magnesium sulfate heptahydrate 132 g / kg, sodium chloride 100.63 g / kg, ferric citrate 29.7 g / kg, manganese sulfate monohydrate 0.8 g / kg, aluminum chloride hexahydrate 0.15 g / kg, zinc sulfate heptahydrate 3 g / kg, potassium iodide 0.15 g / kg, cobalt chloride hexahydrate 1 g / kg, copper chloride 0.1 g / kg, and cellulose 112.61 g / kg.

[0011] The pearl grouper feed material provided by this invention effectively compensates for the deficiencies of unbalanced nutrient ratios and insufficient functional components in existing feeds by adding chromium methionine to the feed, significantly improving nutrient utilization. Experimental results show that this feed can not only promote the protein digestibility and absorption capacity of pearl grouper, enhance antioxidant defense, reduce oxidative stress and improve liver function, but also promote the improvement of the body's health status, thereby providing strong nutritional support for the rapid and healthy growth of the fish and the formation of excellent meat quality. Attached Figure Description

[0012] Figure 1 A cross-section of pearl giant grouper fed with the feed in Comparative Example 1 of the present invention; Figure 2 A cross-section of pearl giant grouper fed with the feed in Comparative Example 2 of the present invention; Figure 3 A slice of pearl giant grouper fed with the feed described in Example 1 of this invention; Figure 4 A slice of pearl giant grouper fed with the feed described in Example 2 of this invention; Figure 5 A slice of pearl giant grouper fed with the feed described in Example 3 of this invention; Figure 6 A slice of pearl giant grouper fed with the feed described in Example 4 of this invention. Detailed Implementation

[0013] The following describes preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications are also considered to be within the scope of protection of the present invention.

[0014] This invention provides a pearl grouper feed, comprising, by weight percentage: 0.01-0.07% chromium methionine, 26-27.5% fish meal, 14-16% soybean meal, 9-10% soybean protein concentrate, 18-19.5% chicken meal, 15.45-20.45% tapioca starch, 0.1-0.12% methionine, 4-4.5% fish oil, 0.9-1.1% vitamin premix, 0.4-0.6% mineral premix, 1.2-1.3% sodium carboxymethyl cellulose, 1.4-1.6% betaine, 0.004-0.006% spermidine, 0.25-0.28% isoleucine, and 0.6-6.0% degummed bone meal.

[0015] Preferably, by weight percentage, the feed comprises: 0.03-0.07% chromium methionine, 26.78% fish meal, 15% soybean meal, 9.45% soybean protein concentrate, 18.78% chicken meal, 15.45-20.45% cassava starch, 0.11% methionine, 4.2% fish oil, 1% vitamin premix, 0.5% mineral premix, 1.25% sodium carboxymethyl cellulose, 1.5% betaine, 0.005% spermidine, 0.26% isoleucine, and 0.665-5.715% degummed bone meal.

[0016] The chromium methionine used in this invention is the commercially available product Amivel. ® Chromium 1000 (Availa) ® The product contains Cr 1000, with a chromium content of 1000~1200mg / kg, a methionine content of ≥0.9%, and a chelation rate of ≥95%, meeting the standards of Q / 320205 KAVN09-2023 to ensure its bioavailability and safety. This product is manufactured by Jinbao (China) Animal Nutrition Technology Co., Ltd.

[0017] Apart from chromium methionine, the other components are conventional feed ingredients in this field, and their selection is a routine choice in aquatic compound feed.

[0018] Chromium methionine is an organic compound formed by the combination of chromium and methionine. It is primarily used to regulate blood sugar, improve insulin sensitivity, and promote metabolic health. In nutrition, it is considered a trace element supplement that helps maintain glucose and lipid metabolism balance by enhancing the bioavailability of chromium. Chromium methionine participates in insulin signaling, helping cells absorb glucose more efficiently. Methionine, as an amino acid carrier, can enhance the absorption rate of chromium, which participates in protein synthesis and fat breakdown. Methionine itself is an important raw material for protein synthesis; the combination of chromium and chromium may synergistically promote muscle repair and reduce body fat accumulation. Methionine contains sulfur groups, which have antioxidant properties and can reduce free radical damage.

[0019] In one specific embodiment of this example, the chromium methionine is added to the basal feed at a ratio of 0.3-0.7 g / kg, and the basal feed has a cassava starch content of 15.45%. In a specific implementation, the chromium methionine can also be added to the basal feed at a ratio of 0.3-0.5 g / kg.

[0020] Preferably, the chromium methionine is added to the basal feed at a ratio of 0.5 to 0.7 g / kg, and the basal feed has a cassava starch content of 15.45%.

[0021] In another specific embodiment of this example, the chromium methionine is added to the high-sugar basal feed at a ratio of 0.5 g / kg, wherein the cassava starch content of the high-sugar basal feed is 20.45%.

[0022] In this invention, the basic feed refers to a feed with a cassava starch content of 15.45%, and the high-sugar basic feed refers to a feed with a cassava starch content of 20.45%. The two have significant differences in cassava starch content and are used to study the breeding environment under normal conditions and high-sugar conditions, respectively, to verify the effect of feed on pearl grouper.

[0023] The pearl grouper feed material provided by this invention effectively compensates for the deficiencies of unbalanced nutrient ratios and insufficient functional components in existing feeds by adding chromium methionine to the feed, significantly improving nutrient utilization. Experimental results show that this feed can not only promote the protein digestibility and absorption capacity of pearl grouper, enhance antioxidant defense, reduce oxidative stress and improve liver function, but also promote the improvement of the body's health status, thereby providing strong nutritional support for the rapid and healthy growth of the fish and the formation of excellent meat quality.

[0024] The preparation method of the pearl grouper feed of the present invention includes the following steps: (1) Crush solid feed ingredients to a particle size that meets the requirements of fish feed standards.

[0025] In practice, the volume of lumpy or granular feed ingredients is reduced and they are crushed into powdered ingredients that meet the particle size requirements of fish feed standards.

[0026] (2) Weigh each solid feed ingredient according to the formula and mix them evenly to form a feed ingredient mixture.

[0027] In practice, to ensure nutritional consistency, the crude protein, crude fat and moisture content of the main raw materials can be measured first, and the weighing ratio can be adjusted accordingly to ensure that the crude protein and crude fat content of the finished feed reaches the target level.

[0028] In a specific embodiment of the present invention, the main feed ingredients include fish meal, chicken meal, soybean protein concentrate and soybean meal, which together account for more than 80% of the total weight of the feed. Therefore, it is of practical significance to measure their nutritional components.

[0029] (3) Stir the feed ingredient mixture for about 30 minutes. During the stirring process, first add the fish oil in the formula, then add water accounting for 30-50% of the total weight of the feed ingredient mixture, and then continue stirring for about 15 minutes.

[0030] (4) The mixed feed mixture is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size.

[0031] In practice, 6) Adjust the feed mold according to the required feed particle size; turn on the feed extruder switch, add the mixed feed ingredients into the feed hopper, and at the same time slowly increase the feed extruder speed.

[0032] (5) The strips are granulated to obtain pellet feed.

[0033] In practice, turn on the pellet mill, put the extruded noodle-shaped feed into the pellet mill, and pellet for 3-5 minutes.

[0034] (6) The pelleted feed is air-dried at room temperature to obtain the finished product.

[0035] To ensure that the feeds of each experimental group are equal in nitrogen and fat, the ratio can be finely adjusted during the formulation process according to the actual nutritional composition of the main protein sources (fish meal, chicken meal, soybean meal, soybean protein concentrate) so that the crude protein content of the finished feed is maintained at 48.66%~48.86% and the crude fat content is maintained at 10.04%~10.13%.

[0036] The method for preparing pearl grouper feed provided by this invention controls the feeding sequence, adding oil first and then water, controlling the amount of water added, and using a room temperature air-drying process. The resulting feed has good water stability and nutrient retention rate, which can not only meet the growth needs of pearl grouper, but also improve the intestinal tissue structure, effectively improve the growth performance of pearl grouper, and the process is simple and suitable for industrial production.

[0037] The pearl grouper feed of the present invention will be described in detail below through specific embodiments.

[0038] To verify the effect of chromium methionine on the growth performance of pearl grouper, this invention prepared several groups of isonitrogenous and isolithic feeds. By setting up a comparative example without chromium methionine and examples with different contents (0.03%, 0.05%, 0.07%) of chromium methionine, the effect of the feed on the growth performance of pearl grouper was comprehensively evaluated.

[0039] The vitamin premixes used in the various embodiments of the present invention (the multivitamins in Table 1) a Prepare per kilogram according to the following proportions: Vitamin B1 2.5g / kg, Vitamin B2 10g / kg, Vitamin B6 2.5g / kg, Vitamin B 120.05g / kg, Vitamin C 14.84g / kg, Vitamin D3 0.0028g / kg, Vitamin E 6.3g / kg, Vitamin K 2g / kg, Niacin 37.5g / kg, Biotin 0.25g / kg, Calcium Pantothenate 25g / kg, Folic Acid 0.75g / kg, Inositol 26.79g / kg, Vitamin A 1g / kg, Fiber 870.5172g / kg.

[0040] The mineral premixes used in the various embodiments of the present invention (compound minerals in Table 1) b Prepare according to the following proportions: Calcium lactate 327 g / kg, potassium chloride 205.28 g / kg, calcium chloride 87.58 g / kg, magnesium sulfate heptahydrate 132 g / kg, sodium chloride 100.63 g / kg, ferric citrate 29.7 g / kg, manganese sulfate monohydrate 0.8 g / kg, aluminum chloride hexahydrate 0.15 g / kg, zinc sulfate heptahydrate 3 g / kg, potassium iodide 0.15 g / kg, cobalt chloride hexahydrate 1 g / kg, copper chloride 0.1 g / kg, cellulose 112.61 g / kg.

[0041] Example 1 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Methionine Chromium 0.03 kg, fish meal 26.78 kg, soybean meal 15 kg, soybean protein concentrate 9.45 kg, chicken meal 18.78 kg, cassava starch 15.45 kg, methionine 0.11 kg, vitamin premix 1 kg, mineral premix 0.5 kg, sodium carboxymethyl cellulose 1.25 kg, betaine 1.5 kg, spermidine 0.005 kg, isoleucine 0.26 kg, degummed bone meal 5.685 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.2 kg of fish oil is added first, followed by 30 kg of water (accounting for 30% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product.

[0042] The total weight of the obtained feed was 100 kg; the crude protein content was 48.74%; and the crude fat content was 10.04%, meeting the requirements of isonitrogenous and islipinous feed.

[0043] Example 2 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Methionine Chromium 0.05 kg, fish meal 26.78 kg, soybean meal 15 kg, soybean protein concentrate 9.45 kg, chicken meal 18.78 kg, cassava starch 15.45 kg, methionine 0.11 kg, vitamin premix 1 kg, mineral premix 0.5 kg, sodium carboxymethyl cellulose 1.25 kg, betaine 1.5 kg, spermidine 0.005 kg, isoleucine 0.26 kg, degummed bone meal 5.665 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.2 kg of fish oil is added first, followed by 30 kg of water (accounting for 30% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product.

[0044] The total weight of the obtained feed was 100 kg; the crude protein content was 48.83%; and the crude fat content was 10.11%, meeting the requirements of isonitrogenous and islipinous feed.

[0045] Example 3 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Chromium methionine 0.05 kg, fish meal 26.78 kg, soybean meal 15 kg, soybean protein concentrate 9.45 kg, chicken meal 18.78 kg, tapioca starch 20.45 kg, methionine 0.11 kg, vitamin premix 1 kg, mineral premix 0.5 kg, sodium carboxymethyl cellulose 1.25 kg, betaine 1.5 kg, spermidine 0.005 kg, isoleucine 0.26 kg, degummed bone meal 0.665 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.2 kg of fish oil is added first, followed by 30 kg of water (accounting for 30% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product.

[0046] The total weight of the obtained feed was 100 kg; the crude protein content was 48.75%; and the crude fat content was 10.07%, meeting the requirements of isonitrogenous and islipinous.

[0047] Example 4 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Methionine Chromium 0.07 kg, fish meal 26.78 kg, soybean meal 15 kg, soybean protein concentrate 9.45 kg, chicken meal 18.78 kg, cassava starch 15.45 kg, methionine 0.11 kg, vitamin premix 1 kg, mineral premix 0.5 kg, sodium carboxymethyl cellulose 1.25 kg, betaine 1.5 kg, spermidine 0.005 kg, isoleucine 0.26 kg, degummed bone meal 5.645 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.2 kg of fish oil is added first, followed by 30 kg of water (accounting for 30% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product.

[0048] The total weight of the obtained feed was 100 kg; the crude protein content was 48.66%; and the crude fat content was 10.12%, meeting the requirements of isonitrogenous and islipinous feed.

[0049] Example 5 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Chromium methionine 0.03 kg, fish meal 27 kg, soybean meal 14.5 kg, soy protein concentrate 10 kg, chicken meal 18.5 kg, tapioca starch 15.45 kg, methionine 0.1 kg, vitamin premix 0.9 kg, mineral premix 0.4 kg, sodium carboxymethyl cellulose 1.2 kg, betaine 1.5 kg, spermidine 0.004 kg, isoleucine 0.25 kg, degummed bone meal 5.666 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.5 kg of fish oil is added first, followed by 30 kg of water (accounting for 30% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product.

[0050] The total weight of the obtained feed was 100 kg; the crude protein content was 48.86%; and the crude fat content was 10.13%, meeting the requirements of isonitrogenous and islipinous feed.

[0051] Example 6 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Chromium methionine 0.07 kg, fish meal 26 kg, soybean meal 14.5 kg, soy protein concentrate 10 kg, chicken meal 19.5 kg, tapioca starch 20.45 kg, methionine 0.1 kg, vitamin premix 0.9 kg, mineral premix 0.4 kg, sodium carboxymethyl cellulose 1.2 kg, betaine 1.4 kg, spermidine 0.004 kg, isoleucine 0.25 kg, degummed bone meal 0.726 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.5 kg of fish oil is added first, followed by 50 kg of water (accounting for 50% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product.

[0052] The total weight of the obtained feed was 100 kg; the crude protein content was 48.78%; and the crude fat content was 10.08%, meeting the requirements of isonitrogenous and islipinous feed.

[0053] Comparative Example 1 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Fish meal 26.78 kg, soybean meal 15 kg, soy protein concentrate 9.45 kg, chicken meal 18.78 kg, tapioca starch 15.45 kg, methionine 0.11 kg, vitamin premix 1 kg, mineral premix 0.5 kg, sodium carboxymethyl cellulose 1.25 kg, betaine 1.5 kg, spermidine 0.005 kg, isoleucine 0.26 kg, degummed bone meal 5.715 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.2 kg of fish oil is added first, followed by 30 kg of water (accounting for 30% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product; The total weight of the obtained feed was 100 kg; the crude protein content was 48.86%; and the crude fat content was 10.08%, meeting the requirements of isonitrogenous and islipinous.

[0054] Comparative Example 2 Crush the following solid feed ingredients to a particle size that meets the requirements of fish feed standards: Fish meal 26.78 kg, soybean meal 15 kg, soybean protein concentrate 9.45 kg, chicken meal 18.78 kg, tapioca starch 20.45 kg, methionine 0.11 kg, vitamin premix 1 kg, mineral premix 0.5 kg, sodium carboxymethyl cellulose 1.25 kg, betaine 1.5 kg, spermidine 0.005 kg, isoleucine 0.26 kg, degummed bone meal 0.715 kg; Mix the above-mentioned pulverized solid feed ingredients evenly to form a feed ingredient mixture; The feed ingredient mixture is stirred. During the stirring process, 4.2 kg of fish oil is added first, followed by 30 kg of water (accounting for 30% of the total weight of the feed ingredient mixture), and then stirring is continued. The mixed feed is extruded into strips, wherein feed molds are pre-configured according to the target feed particle size. The strip-shaped material is granulated to obtain pelleted feed; The pelleted feed is air-dried at room temperature to obtain the finished product; The total weight of the obtained feed was 100 kg; the crude protein content was 48.78%; and the crude fat content was 10.13%, meeting the requirements of isonitrogenous and islipinous feed.

[0055] The feeds prepared in Comparative Examples 1, 2, and Examples 1-4 correspond to groups A0, A1, A2, A3, A4, and A5 in Table 1 below. Group A0 is the control group, consisting of a basal feed without added methionine chromium. Comparative Example 1 corresponds to Group A0 (basic feed, without added chromium methionine). Comparative Example 1 corresponds to Group A1 (high-sugar basal diet, without added methionine chromium). Example 1 corresponds to group A2 (basic feed, supplemented with 0.03% chromium methionine). Example 2 corresponds to group A3 (basic feed, supplemented with 0.05% chromium methionine); Example 3 corresponds to group A4 (high sugar basal feed, with 0.05% methionine chromium added); Example 4 corresponds to group A5 (basic feed, with 0.07% methionine chromium added).

[0056] All groups met the isonitrogenous and islipinous requirements (crude protein content 48.66%–48.86%, crude fat content 10.04%–10.13%), and could be used in aquaculture trials to evaluate the effectiveness of chromium methionine in different carbohydrate backgrounds.

[0057] Table 1 Feed formulations for comparative examples and embodiments Experimental Example a. Feeding process The rearing experiment was conducted in indoor self-circulating glass tanks (60cm x 48cm x 60cm). Before formal rearing, the pearl grouper were temporarily housed in the glass tanks to acclimatize. After eight weeks of temporary housing, the formal rearing experiment began. The experiment used 432 pearl grouper (initial weight: 93±1g). After a 24-hour starvation period, they were randomly assigned to 24 glass tanks, with 18 pearl grouper per tank, forming 6 groups (corresponding to diets A0-A5), with 4 replicates per group.

[0058] During the breeding period, the animals were fed twice a day (8:00 and 16:30) until full. The breeding experiment lasted for 8 weeks, with a water temperature of 26~28℃. The dissolved oxygen in the breeding water was >6mg / L, ammonia nitrogen <0.2mg / L, and nitrite <0.05mg / L.

[0059] b. Sample collection At the end of the experiment, the fish were fasted for 24 hours before sampling. Nine fish were randomly selected from each tank and collected after anesthesia with MS-222 (0.1 g / L, Sigma Aldrich, USA). One fish was used for the preparation of intestinal and liver tissue sections; three fish were anesthetized and stored on ice (-20℃) for whole-fish composition analysis; the remaining five fish were used to collect blood, muscle, heart, head kidney, gallbladder, brain, liver and intestine in sequence, and the weights of the visceral mass, intestine, liver and mesenteric fat were recorded at the same time.

[0060] Two blood samples were taken from each fish. One half was added with sodium heparin and centrifuged for half an hour (4°C, 2000-3000g, 25min) to obtain a plasma sample. The other half was left to stand overnight and then centrifuged (4°C, 2000-3000g, 25min) to obtain a serum sample. All tissue samples, including those from the brain, liver, and intestines, were rapidly transferred to liquid nitrogen and subsequently stored in an ultra-low temperature freezer at -80°C.

[0061] c. Sample Analysis (i) Crude component analysis The analysis of routine nutritional components of feed and fish carcasses in whole fish, whole muscle, and whole liver samples followed the methods of AOAC (1995). Crude protein content was determined using the Dumas combustion method (Rapid Max N exceed, Elementar, Germany); crude fat content was determined using the Soxhlet extraction method (XT15i, ANKOM, USA); and moisture content was determined using both drying and freeze-drying methods.

[0062] (ii) Enzyme activity detection Samples from each tissue were removed from a -80 ℃ freezer and thawed before being tested for different enzyme activities according to the kit instructions. During the tissue enzyme activity assay, an appropriate amount of pearl grouper tissue was taken and pretreated according to the corresponding kit instructions. Specifically, the tissue was accurately weighed and homogenized at a ratio of 1:9 (weight (g) to physiological saline (mL)); the supernatant was then collected by centrifugation for subsequent measurements. After the preliminary experiment was completed, the formal batch experiments were conducted.

[0063] (iii) Observation of organizational morphology Liver tissue fixed in paraformaldehyde solution was fixed, dehydrated, embedded, sectioned, dewaxed, and stained. Following differentiation, washing, and bluing, the tissue was finally dehydrated, cleared, and mounted. The prepared sections were then observed and images acquired using Saiviewer software.

[0064] d. Data Analysis The formulas for calculating growth performance evaluation indicators are as follows: Survival rate (SR%) = Number of survivors / Total number of survivors × 100; Weight gain rate (WGR%) = (final weight - initial weight) / initial weight × 100; Specific growth rate (SGR%) / d = (ln final body weight - ln initial body weight) / number of days of rearing × 100; Feed conversion ratio (FCR) = Feed consumption / Weight gain; Hepatosomatic index (HSI%) = (weight of digestive glands / body weight) × 100; Conditioniness (CF%) = Final total weight / Body length cubed × 100%.

[0065] Protein efficiency (PER%) = (Final total weight - Initial total weight) / Feed protein intake × 100%.

[0066] Protein Production Value (PRR%) = (Final Body Protein - Initial Body Protein) / Feed Protein Intake × 100%.

[0067] Lipid deposition rate (LRR%) = (final body fat - initial body fat) / dietary fat intake × 100%.

[0068] Experimental data were analyzed using SPSS 227.0 software (IBM, USA) for one-way ANOVA. Multiple comparisons between groups were performed using the Tucky method. The significance level was P < 0.05. The results are expressed as mean ± standard deviation.

[0069] e. Experimental Results (i) Growth performance and morphological indicators The growth performance of pearl grouper after being fed with diets containing different levels of methionine and chromium was shown in Table 2 below.

[0070] Table 2. Effects of methionine and chromium addition on growth performance of pearl grouper. Note: Values ​​in the same row without letters or with the same superscript indicate no statistically significant difference (P>0.05), while different superscripts indicate a statistically significant difference (P<0.05). The same applies to the table below.

[0071] It can be seen that with the increase of chromium methionine concentration, the growth performance, feed utilization efficiency, protein deposition capacity and body condition of fish are significantly improved.

[0072] Compared with the control groups A0 and A1 without chromium methionine, the A2-A5 groups with chromium methionine generally performed better in terms of final weight, weight gain rate (WGR), specific growth rate (SGR), feed conversion ratio (FCR), protein efficiency (PER), protein deposition rate (PRR), and condition factor (CF), and most of the differences were statistically significant (P<0.05).

[0073] Among them, groups A4 and A5 achieved higher levels in many key indicators, significantly outperforming groups A0 and A1. Group A5 showed a slight decline in a few indicators compared to group A4 (possibly due to a slight plateau or feedback adjustment caused by excessively high concentrations), but was still significantly better than the low-concentration groups (groups A2 and A3) and the control groups A0 and A1.

[0074] It is worth noting that Group A4 achieved peak performance in almost all key indicators (highest in final weight, WGR, SGR, FCR, PER, PRR, and CF), demonstrating the best growth performance and feed efficiency.

[0075] The high-sugar control group (A1) showed significantly lower final weight, wgg growth rate (WGR), and sigma growth rate (SGR) compared to other groups, indicating that simply increasing the proportion of cassava starch without the addition of chromium methionine inhibited fish growth. However, the addition of chromium methionine (such as in group A4) significantly improved fish growth performance even under high-sugar conditions.

[0076] As can be seen from the above, chromium methionine can effectively promote growth, reduce feed conversion ratio, improve nutrient utilization, and maintain 100% survival rate, making it a highly efficient and safe growth promoter.

[0077] (ii) Promotes the activity of healthy enzymes in the body Table 3 shows the effects of chromium methionine on the activity of enzymes related to the overall health of pearl grouper. Table 3. Effects of chromium methionine on the activity of healthy enzymes in pearl grouper. Note: Values ​​in the same row without letters or with the same superscript indicate no statistically significant difference (P>0.05), while different superscripts indicate a statistically significant difference (P<0.05). The same applies to the table below.

[0078] As can be seen from the experimental results in Table 3, with the increase of chromium methionine concentration, the additive showed a significant dose-dependent positive effect on the digestive capacity, intestinal health, antioxidant capacity and liver protection of pearl grouper.

[0079] The effect is more obvious when the concentration is moderately increased (usually reaching the peak in the A3–A4 group). At excessively high concentrations (such as the A5 group), some indicators may decline slightly, but they are still significantly better than those at low concentrations or the control group.

[0080] This indicates that the additive promotes protein digestion, enhances antioxidant defense, reduces oxidative stress, and improves liver function, making it a potential positive feed optimizer.

[0081] Among them, most indicators in groups A3 to A4 reached their peak (high digestive enzymes, strong antioxidants, low liver damage, and lowest oxidation products), indicating that at this level of chromium methionine addition, it can maximize the promotion of digestion and absorption, enhance antioxidant and immune functions, and protect liver and intestinal health, thereby potentially improving growth performance and disease resistance.

[0082] Some indicators in group A5 (such as SOD and amylase) were slightly lower than those in group A4, but it still maintained good effects in liver protection and MDA reduction, indicating that it was well tolerated at this concentration and had no obvious negative effects.

[0083] Compared with the control group, the high sugar group (A1) performed worse in several indicators (such as weak antioxidant, high liver enzyme, and elevated MDA level), indicating that simply increasing the proportion of sugar (cassava starch) in the feed without adding methionine chromium will have a negative impact on the health of fish.

[0084] (iii) Histological observation Intestinal tissue morphology of pearl grouper, such as Figures 1-6 (As shown in Comparative Example 1, Comparative Example 2, Example 1, Example 2, Example 3, and Example 4 respectively), compared with the control group, the intestinal morphology of the pearl grouper in the experimental group with added chromium methionine was relatively intact. The intestinal villi of the pearl grouper in the group with added chromium methionine were more regularly arranged and more tightly, the surface folds were more developed, the connective tissue was more compact, and the muscle bundles wrapped around it were more intact. The above morphological structure diagrams show that the number and length of intestinal villi increased, and the intestinal wall thickened, which helps to improve the absorption of nutrients in the feed.

[0085] (iv) The safety of eating pearl grouper The amino acid chromium level in feed has a significant promoting effect on the growth performance of pearl grouper, but food safety issues cannot be ignored. Excessive chromium may lead to poisoning. my country's food safety standards stipulate that the chromium content in food should be less than 2.0 mg / kg. Therefore, the feed samples of this invention were commissioned to a qualified third-party testing institution for chromium content testing. The lowest detection limit was 0.15 mg / kg. The results showed that chromium was not detected in any samples, indicating that the chromium content of this product is less than 0.15 mg / kg, which meets the national food safety standards.

[0086] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A feed for pearl grouper, characterized in that, By weight percentage, it includes: 0.01-0.07% chromium methionine, 26-27.5% fish meal, 14-16% soybean meal, 9-10% soy protein concentrate, 18-19.5% chicken meal, 15.45-20.45% tapioca starch, 0.1-0.12% methionine, 4-4.5% fish oil, 0.9-1.1% vitamin premix, 0.4-0.6% mineral premix, 1.2-1.3% sodium carboxymethyl cellulose, 1.4-1.6% betaine, 0.004-0.006% spermidine, 0.25-0.28% isoleucine, and 0.6-6.0% degummed bone meal.

2. The pearl grouper feed according to claim 1, characterized in that, By weight percentage, it includes: 0.03-0.07% chromium methionine, 26.78% fish meal, 15% soybean meal, 9.45% soy protein concentrate, 18.78% chicken meal, 15.45-20.45% tapioca starch, 0.11% methionine, 4.2% fish oil, 1% vitamin premix, 0.5% mineral premix, 1.25% sodium carboxymethyl cellulose, 1.5% betaine, 0.005% spermidine, 0.26% isoleucine, and 0.665-5.715% degummed bone meal.

3. The pearl grouper feed according to claim 1, characterized in that, The chromium methionine is added to the basal feed at a ratio of 0.3 to 0.7 g / kg, and the basal feed has a cassava starch content of 15.45%.

4. The pearl grouper feed according to claim 3, characterized in that, The chromium methionine is added to the basal feed at a ratio of 0.5 to 0.7 g / kg.

5. The pearl grouper feed according to claim 1, characterized in that, The chromium methionine is added to the high-sugar basal feed at a ratio of 0.5 g / kg, wherein the cassava starch content of the high-sugar basal feed is 20.45%.

6. The pearl grouper feed according to claim 1, characterized in that, The vitamin premix comprises the following components per kilogram of premix: vitamin B1 2.5 g / kg, vitamin B2 10 g / kg, vitamin B6 2.5 g / kg, vitamin B... 12 0.05g / kg, Vitamin C 14.84g / kg, Vitamin D3 0.0028g / kg, Vitamin E 6.3g / kg, Vitamin K 2g / kg, Niacin 37.5g / kg, Biotin 0.25g / kg, Calcium Pantothenate 25g / kg, Folic Acid 0.75g / kg, Inositol 26.79g / kg, Vitamin A 1g / kg, Fiber 870.5172g / kg.

7. The pearl grouper feed according to claim 1, characterized in that, The mineral premix comprises the following components per kilogram: calcium lactate 327 g / kg, potassium chloride 205.28 g / kg, calcium chloride 87.58 g / kg, magnesium sulfate heptahydrate 132 g / kg, sodium chloride 100.63 g / kg, ferric citrate 29.7 g / kg, manganese sulfate monohydrate 0.8 g / kg, aluminum chloride hexahydrate 0.15 g / kg, zinc sulfate heptahydrate 3 g / kg, potassium iodide 0.15 g / kg, cobalt chloride hexahydrate 1 g / kg, copper chloride 0.1 g / kg, and cellulose 112.61 g / kg.

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