HYDROLYZED COMPOUND PROTEIN FOR LARVAL LARGEMOUTH BASS (MICROPTERUS SALMOIDES) AND USE THEREOF

Abstract

A hydrolysate compound protein for larval largemouth bass (Micropterus salmoides) and use thereof for aquatic animal feeds. The micro-diet was formulated with the following components: 72-74 parts by weight of a hydrolyzed protein pre-mixture, 3-5 parts by weight of mixed fish soluble meal, 4-6 parts by weight of gluten flour, 2-4 parts by weight of sodium caseinate, and 0.4-0.6 parts by weight of a yeast hydrolysate, where the hydrolyzed protein pre-mixture includes white fish meal, fermented soybean meal, shrimp meal and blood meal with the weight ratio of 58-62: 5-7: 4-6: 2-3. The compound protein is obtained by fully pulverizing the foregoing components, passing through a 100-mesh sieve, fully mixing in proportion, and being hydrolyzed with neutral protease and keratinase.

Claims

1. A compound protein for larval largemouth bass (Micropterus salmoides), comprising the following components: 72-74 parts by weight of a hydrolyzed protein pre-mixture, 3-5 parts by weight of mixed fish soluble meal, 4-6 parts by weight of gluten flour, 2-4 parts by weight of sodium caseinate, and 0.4-0.6 parts by weight of a yeast hydrolysate, wherein the compound protein for larval largemouth bass is obtained by fully pulverizing the foregoing components, passing through a 100-mesh sieve, and fully mixing in proportion, and the hydrolyzed protein pre-mixture comprises white fish meal, fermented soybean meal, shrimp meal and blood meal, and the weight ratio is 58-62: 5-7: 4-6: 2-3, respectively.

2. The compound protein for larval largemouth bass according to claim 1, wherein the compound protein for larval largemouth bass comprises the following components: 73 parts by weight of the hydrolyzed protein pre-mixture, 4 parts by weight of the mixed fish soluble meal, 5 parts by weight of the gluten flour, 3 parts by weight of the sodium caseinate, and 0.5 parts by weight of the yeast hydrolysate, wherein the hydrolyzed protein pre-mixture comprises white fish meal, fermented soybean meal, shrimp meal and blood meal with the weight ratio of 60: 6: 5: 2.

3. The compound protein for larval largemouth bass according to claim 1, wherein the hydrolyzed protein pre-mixture is prepared by a method comprising the following steps: step 1, premixing the white fish meal, the fermented soybean meal, the shrimp meal, and the blood meal to obtain a premix, and fully mixing the premix with sterile distilled water in a solid-liquid ratio of 1:2; step 2, weighing 2 wt% neutral protease in the premix and 1 wt% keratinase in the premix, mixing evenly, and slowly adding a mixture obtained in step 1 while stirring; step 3, heating a mixture in step 2 in a 50° C. water bath for 30 min, and stirring the mixture once every 10 min; step 4, putting an enzymatic hydrolysate in step 3 into an air-drying oven at 105° C. for sterilization and enzyme inactivation for 30 min; and step 5, keeping drying a product after the sterilization and the enzyme inactivation in step 4 at 80° C. for 12 h, and obtaining the hydrolyzed protein pre-mixture.

4. The compound protein for larval largemouth bass according to claim 2, wherein the hydrolyzed protein pre-mixture is prepared by a method comprising the following steps: step 1, premixing the white fish meal, the fermented soybean meal, the shrimp meal, and the blood meal to obtain a premix, and fully mixing the premix with sterile distilled water in a solid-liquid ratio of 1:2; step 2, weighing 2 wt% neutral protease in the premix and 1 wt% keratinase in the premix, mixing evenly, and slowly adding a mixture obtained in step 1 while stirring; step 3, heating a mixture in step 2 in a 50° C. water bath for 30 min, and stirring the mixture once every 10 min; step 4, putting an enzymatic hydrolysate in step 3 into an air drying oven at 105° C. for sterilization and enzyme inactivation for 30 min; and step 5, keeping drying a product after the sterilization and the enzyme inactivation in step 4 at 80° C. for 12 h, and obtaining the hydrolyzed protein pre-mixture.

5. A micropellet feed for larval largemouth bass, comprising the compound protein for larval largemouth bass of claim 1.

6. Use of the compound protein for larval largemouth bass according to claim 1 in promoting the growth, intestinal development and reduction of deformity rate of larval largemouth bass.

7. The use of according to claim 6, wherein the compound protein for larval largemouth bass comprises the following components: 73 parts by weight of the hydrolyzed protein pre-mixture, 4 parts by weight of the mixed fish soluble meal, 5 parts by weight of the gluten flour, 3 parts by weight of the sodium caseinate, and 0.5 parts by weight of the yeast hydrolysate, wherein the hydrolyzed protein pre-mixture comprises white fish meal, fermented soybean meal, shrimp meal and blood meal with the weight ratio of 60: 6: 5: 2..

8. The use of according to claim 6, wherein the hydrolyzed protein pre-mixture is prepared by a method comprising the following steps: step 1, premixing the white fish meal, the fermented soybean meal, the shrimp meal, and the blood meal to obtain a premix, and fully mixing the premix with sterile distilled water in a solid-liquid ratio of 1:2; step 2, weighing 2 wt% neutral protease in the premix and 1 wt% keratinase in the premix, mixing evenly, and slowly adding a mixture obtained in step 1 while stirring; step 3, heating a mixture in step 2 in a 50° C. water bath for 30 min, and stirring the mixture once every 10 min; step 4, putting an enzymatic hydrolysate in step 3 into an air-drying oven at 105° C. for sterilization and enzyme inactivation for 30 min; and step 5, keeping drying a product after the sterilization and the enzyme inactivation in step 4 at 80° C. for 12 h, and obtaining the hydrolyzed protein pre-mixture.

9. The use of according to claim 7, wherein the hydrolyzed protein pre-mixture is prepared by a method comprising the following steps: step 1, premixing the white fish meal, the fermented soybean meal, the shrimp meal, and the blood meal to obtain a premix, and fully mixing the premix with sterile distilled water in a solid-liquid ratio of 1:2; step 2, weighing 2 wt% neutral protease in the premix and 1 wt% keratinase in the premix, mixing evenly, and slowly adding a mixture obtained in step 1 while stirring; step 3, heating a mixture in step 2 in a 50° C. water bath for 30 min, and stirring the mixture once every 10 min; step 4, putting an enzymatic hydrolysate in step 3 into an air drying oven at 105° C. for sterilization and enzyme inactivation for 30 min; and step 5, keeping drying a product after the sterilization and the enzyme inactivation in step 4 at 80° C. for 12 h, and obtaining the hydrolyzed protein pre-mixture.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0020] In order to make the objectives, technical solutions and advantages of the examples of the present disclosure clearer, the technical solutions of the examples of the present disclosure will be described clearly and completely below with reference to the examples of the present disclosure. Apparently, the described examples are a part of, but not all of, the examples of the present disclosure. Based on the described examples of the present disclosure, all other examples obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present disclosure.

[0021] The present disclosure relates to a compound protein for larval largemouth bass, including the following components: 72-74 parts by weight of a hydrolyzed protein pre-mixture, 3-5 parts by weight of mixed fish soluble meal, 4-6 parts by weight of gluten flour, 2-4 parts by weight of sodium caseinate, and 0.4-0.6 parts by weight of a yeast hydrolysate, where the compound protein for larval largemouth bass is obtained by fully pulverizing the foregoing components, passing through a 100-mesh sieve, and fully mixing in proportion, and the hydrolyzed protein pre-mixture includes 58-62 parts by weight of white fish meal, 5-7 parts by weight of fermented soybean meal, 4-6 parts by weight of shrimp meal, and 2-3 parts by weight of blood meal.

[0022] According to some embodiments, the compound protein for larval largemouth bass includes the following components: 73 parts by weight of the hydrolyzed protein pre-mixture, 4 parts by weight of the mixed fish soluble meal, 5 parts by weight of the gluten flour, 3 parts by weight of the sodium caseinate, and 0.5 parts by weight of the yeast hydrolysate, where the hydrolyzed protein pre-mixture includes 60 parts by weight of the white fish meal, 6 parts by weight of the fermented soybean meal, 5 parts by weight of the shrimp meal, and 2 parts by weight of the blood meal.

[0023] The present disclosure further relates to a micro-diet for larval largemouth bass, containing the foregoing compound protein for larval largemouth bass.

[0024] In the following examples, the hydrolyzed protein pre-mixture may be prepared by a method containing the following steps: [0025] step 1, premixing the white fish meal, the fermented soybean meal, the shrimp meal, and the blood meal to obtain a premix, and fully mixing the premix with sterile distilled water in a solid-liquid ratio of 1:2; [0026] step 2, weighing 2 wt% neutral protease in the premix and 1 wt% keratinase in the premix, mixing evenly, and slowly adding a mixture obtained in step 1 while stirring; [0027] step 3, heating a mixture in step 2 in a 50° C. water bath for 30 min, and stirring the mixture once every 10 min; [0028] step 4, putting an enzymatic hydrolysate into an air-drying oven at 105° C. for sterilization and enzyme inactivation for 30 min; and [0029] step 5, keeping drying a product after the sterilization and the enzyme inactivation at 80° C. for 12 h, and obtaining the hydrolyzed protein pre-mixture.

[0030] The present disclosure will be further explained and described below with reference to specific examples.

Example 1

[0031] In this example, a compound protein for larval largemouth bass was prepared. First, a hydrolyzed protein pre-mixture was prepared according to the foregoing method, fully pulverized with mixed fish soluble meal, gluten flour, sodium caseinate, and yeast hydrolysate, and passed through a 100-mesh sieve; 7.2 kg of hydrolyzed protein pre-mixture, 0.3 kg of mixed fish soluble meal, 0.4 kg of gluten flour, 0.2 kg of sodium caseinate, and 0.04 kg of yeast hydrolysate were weighed, and all components were mixed uniformly step by step (in ascending order by weight).

Example 2

[0032] In this example, a compound protein for larval largemouth bass was prepared. First, a hydrolyzed protein pre-mixture was prepared according to the foregoing method, fully pulverized with mixed fish soluble meal, gluten flour, sodium caseinate, and yeast hydrolysate, and passed through a 100-mesh sieve; 7.4 kg of hydrolyzed protein pre-mixture, 0.5 kg of mixed fish soluble meal, 0.6 kg of gluten flour, 0.4 kg of sodium caseinate, and 0.06 kg of yeast hydrolysate were weighed, and all components were mixed uniformly step by step (in ascending order by weight).

Example 3

[0033] In this example, a compound protein for larval largemouth bass was prepared. First, a hydrolyzed protein pre-mixture was prepared according to the foregoing method, fully pulverized with mixed fish soluble meal, gluten flour, sodium caseinate, and yeast hydrolysate, and passed through a 100-mesh sieve; 7.3 kg of hydrolyzed protein pre-mixture, 0.4 kg of mixed fish soluble meal, 0.5 kg of gluten flour, 0.3 kg of sodium caseinate, and 0.05 kg of yeast hydrolysate were weighed, and all components were mixed uniformly step by step (in ascending order by weight).

Example 4 Detection of Aquaculture Effect

[0034] The compound protein (the premixed protein was not treated by enzymolysis, and the proportion of each component was the same as that of the compound protein provided by the present disclosure) was used as a positive control, and the compound protein for larval largemouth bass in Example 3 was used as an experimental group; other components in the feed preparation process except the hydrolyzed protein pre-mixture feed were consistent with those of commercial feeds. The specific production steps were as follows: 8.55 kg of the foregoing compound protein, 0.3 kg of starch, 0.12 kg of soybean oil, 0.5 kg of soybean lecithin powder, 0.13 kg of vitamin premix, 0.10 kg of mineral premix, and 0.10 kg of calcium dihydrogen phosphate were weighed and fully mixed to obtain a micro-diet for larval largemouth bass.

[0035] The implementation site was located in the joint laboratory of Shanghai Ocean University and Guangdong Evergreen Feed Industry Co., Ltd. Using the current commercial feed as a negative control, and the larval largemouth bass 17 days after hatching as breeding subjects, a 4-week aquaculture experiment was carried out. The larval largemouth bass were fed 4 times a day. Three replicates were done for each treatment, and there were 2,500 fries per barrel.

[0036] The experimental data were expressed as mean ± standard error (X ± SEM). The experimental results were analyzed by one-way analysis of variance (ANOVA) using SPSS17.0 software. Tukey’s multiple range test was chosen as a multiple comparison test and the significance level of 5% was used. The same letters following the data in the table indicate that insignificant differences.

[0037] The results showed that, compared with the control group, the compound protein for larval largemouth bass obtained in Example 3 significantly increased the final body length, final body weight and survival rate of the larval largemouth bass, and also substantially reduced the deformity rate of the larval largemouth bass; meanwhile, the above indexes in the positive control group were significantly better than those obtained by the commercial feed (negative control group) (Table 1).

[0038] Digestive system underdevelopment is a major feature of larvae, and the level of digestive enzyme activity can often effectively evaluate the development of their digestive system. This example further proved that the compound protein for larval largemouth bass in Example 3 significantly elevated the activity of pepsin, trypsin and alkaline phosphatase in the larval largemouth bass, and significantly reduced the activity of amylase, demonstrating that the compound protein for larval largemouth bass provided by the present disclosure can significantly promote the development of the digestive system. Meanwhile, the above indexes of the positive control group were significantly better than those obtained by the commercial feed (negative control group) (Table 2).

TABLE-US-00001 Effects of the use of the compound protein on the growth indexes of larval largemouth bass Negative control group Positive control group Experimental group Initial body weight (mg) 9.50±0.01 9.50±0.01 9.50±0.01 Initial body length (mm) 9.61±0.01 9.61±0.01 9.61±0.01 Final body weight (g) 0.68±0.00.sup.c 1.18±0.09.sup.b 1.43±0.18.sup.a Final body length (cm) 2.82±0.04.sup.c 3.94±0.05.sup.b 4.28±0.03.sup.a Specific growth rate 15.25±0.20.sup.c 18.56±0.16.sup.b 19.30±0.12.sup.a Deformity rate (%) 41.29±1.05.sup.c 23.01±1.94.sup.b 0.48±0.29.sup.a Survival rate (%) 15.95±0.80.sup.c 29.27±1.18.sup.b 35.59±1.69.sup.a

TABLE-US-00002 Effects of the use of the compound protein on the activity of digestive enzymes of larval largemouth bass Negative control group Positive control group Experimental group Alkaline phosphatase (U/mgprot) 0.56±0.02.sup.c 0.78±0.03.sup.b 1.01±0.02.sup.a Pepsin (U/mgprot) 1.46±0.01.sup.c 1.96±0.03.sup.b 2.67±0.10.sup.a Amylase (U/gprot) 36.14±0.41.sup.a 25.44±0.98.sup.b 19.61±1.70.sup.c Trypsin (U/mgprot) 515.72±12.31.sup.c 859.76±15.80.sup.b 1851.92±48.75.sup.a

[0039] To sum up, the use of the compound protein for larval largemouth bass provided by the present disclosure in the micro-diet can significantly improve the growth performance and survival rate of larval largemouth bass, promote the development of the digestive system, and significantly reduce the deformity rate. Using the compound proteins for larval largemouth bass of Example 1 and Example 2 also obtains similar results to Example 3, and the intragroup parallel test also rules out the operating error.