CN118303345A - Nutritional method for improving anti-hypoxia stress capability of large yellow croaker - Google Patents

Nutritional method for improving anti-hypoxia stress capability of large yellow croaker Download PDF

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CN118303345A
CN118303345A CN202410358921.9A CN202410358921A CN118303345A CN 118303345 A CN118303345 A CN 118303345A CN 202410358921 A CN202410358921 A CN 202410358921A CN 118303345 A CN118303345 A CN 118303345A
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large yellow
yellow croaker
feed
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张文兵
章程伟
曲朋
李昕昕
秦高婵
姜慧
张远明
麦康森
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Ocean University of China
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    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish
    • Y02A40/818—Alternative feeds for fish, e.g. in aquacultures

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Abstract

The invention provides a nutritional method for improving the anti-hypoxia stress capability of large yellow croaker, and belongs to the technical field of aquaculture. The method comprises the following steps: preparing, processing, feeding and breeding feed. The feed is prepared from clostridium ethoxide protein, cotton seed concentrated protein, kelp powder containing gelatin and 5-aminolevulinic acid as main components of the compound feed for feeding the large yellow croaker, improves the proportion of fish meal, reduces the negative influence on the health of the large yellow croaker body, and simultaneously combines a specific processing technology and a feed feeding mode to improve the survival rate, the oxidation resistance and the hypoxia stress resistance of the large yellow croaker. In addition, the nutritional method for improving the anti-hypoxia stress capability of the large yellow croaker has the characteristics of small feed consumption, low production cost and stable effect.

Description

Nutritional method for improving anti-hypoxia stress capability of large yellow croaker
Technical Field
The invention relates to the technical field of aquaculture, in particular to a nutritional method for improving the anti-hypoxia stress capability of large yellow croakers.
Background
Large yellow croaker (LARIMICHTHY SCROCEA) is the sea water economic fish with the highest culture yield in China, and is called as sea water national fish. The wild large yellow croaker has golden body color, delicious taste and higher nutritive value, and is deeply favored by consumers. In recent years, the yield of large yellow croakers in China is increased year by year, and the yield is increased from 8 ten thousand tons in 2011 to 25 ten thousand tons in 2020. Large yellow croaker is sea water carnivorous fish, the proportion of fish meal in the compound feed is relatively large, and the shortage of fish meal and the excessively high proportion of feed cost in the cultivation cost are problems facing the global aquaculture industry. Clostridium ethoxide protein (Clostridium autoethanogenumprotein, CAP) is a byproduct of ethanol production by carbon monoxide waste gas of the steel industry. As a novel protein source, the protein source is rich in nutrition, contains up to 80% of protein, and does not contain anti-nutritional factors in plant proteins. China is the country with the largest cotton yield in the world, and cotton seed resources are very rich. The cotton seed concentrated protein (Cottonseedprotein concentrate, CPC) adopts a more perfect process relative to cotton seed meal, maintains the nutritive value by reducing the thermal denaturation of protein, and more thoroughly removes anti-nutritional factors such as gossypol and the like. The novel protein source has the property of non-grain, has the characteristics of high protein content and relatively balanced amino acid, is relatively low in price, and plays an important role in relieving the problem of fish meal shortage. The novel protein source is used for replacing fish meal, so that the feed cost can be reduced, the shortage of the fish meal is relieved, but the reduction of the fish meal content in the feed can cause a series of negative effects on the growth performance, oxidation resistance, immunity and other aspects of the large yellow croaker, and the large yellow croaker usually faces a certain degree of anoxic stress in the process of fishing and transporting, so that the fish body is damaged and even dies, and economic loss is caused.
5-Aminolevulinic acid (Aminolevulinic Acid), also known as delta-aminolevulinic acid, abbreviated as 5-ALA, is an important precursor for biosynthesis of tetrahydropyrrole, and is an essential substance for biosynthesis of heme, cytochrome, vitamin B12 and chlorophyll. The 5-ALA can improve the oxygen transmission, nutrient transmission and metabolism efficiency in organisms by activating erythrocyte and myoglobin in the animals, thereby improving the utilization rate of the feed, increasing the absorption rate and the metabolism speed and having obvious physiological effects. The daily ration supplement of 5-ALA can promote synthesis of heme, increase concentration of hemoglobin and iron ions and other blood components, and improve immune response, which is especially important for young livestock and livestock with high production load, the immune system of which is not yet developed and mature. The kelp powder (Kelp powder) containing gelatin has the main active components of algin, and the algin has the functions of promoting animal growth, resisting oxidation and regulating immunity.
Therefore, the method has very important production significance in relieving the problems of reduced growth performance, oxidation resistance and immunity caused by reduced fish meal content, enhancing the tolerance of fish bodies in low-oxygen environment and reducing death rate.
Disclosure of Invention
In view of this, the present invention provides a nutritional method for improving the anti-hypoxia stress capacity of large yellow croaker, comprising the steps of: comprises the steps of preparing, processing, feeding and breeding feed, and also comprises the steps of mixing clostridium ethoxide protein, cotton seed concentrated protein, kelp powder containing gelatin and 5-aminolevulinic acid with basic feed, and then carrying out puffing granulation, drying and post-spraying to obtain the large yellow croaker floatability compound feed; the floating compound feed is put into a culture pond of large yellow croaker, and is fed twice a day, wherein the time interval between the two times is 12-13 hours.
Preferably, the addition amount of the 5-aminolevulinic acid in the feed is 2-162 mg/kg.
Further preferably, the addition amount of the 5-aminolevulinic acid in the feed is 18-162 mg/kg.
Preferably, the ingredients of the basic feed comprise: fish meal, herborist peptide, tapioca starch, standard flour, fish oil, soybean lecithin, premix, choline chloride, monocalcium phosphate, mildew inhibitor, antioxidant, ethoxyquinoline, microcrystalline cellulose and phagostimulant.
Preferably, the premix comprises ：FeSO4·H2O,ZnSO4·H2O,NaCl,MnSO4·H2O,CuSO4·5H2O,CoCl2·6H2O,Na2SeO3, potassium iodate and zeolite powder.
Preferably, the nutritional method for improving the anti-hypoxia stress capability of the large yellow croaker, and the preparation method of the floating compound feed comprise the following steps:
(1) 0.5 to 2 weight percent of premix, 0.01 to 0.02 weight percent of antioxidant, 0.05 to 0.1 weight percent of mildew inhibitor, 0.15 to 0.3 weight percent of phagostimulant, 0.1 to 0.3 weight percent of choline chloride, 0.8 to 2 weight percent of monocalcium phosphate and 0.5 to 1.5 weight percent of kelp powder containing glue are weighed and mixed uniformly according to the mass ratio to obtain a mixture A, and 5-ALA is added for standby;
(2) Weighing and uniformly mixing 30-40 wt% of fish meal, 7.14-10.08 wt% of clostridium ethoxide protein, 18.26-25.76 wt% of cottonseed concentrated protein, 4-5 wt% of Baiwei peptide, 4-5 wt% of tapioca starch, 10-12 wt% of standard flour, 0.03-0.06 wt% of ethoxyquinoline and 2.29-3.38wt% of microcrystalline cellulose according to a mass ratio to obtain a mixture B for standby;
(3) Mixing the mixture A and the mixture B, and sequentially puffing, granulating and drying to obtain a semi-finished product of the compound feed;
(4) Uniformly mixing 3.9-6wt% of fish oil, 1-3wt% of soybean oil and 1-1.5wt% of soybean lecithin according to a mass ratio to obtain an oil mixture, and then spraying the oil mixture on the surface of a semi-finished product of the compound feed to obtain a finished product of the floatable compound feed.
Preferably, the process parameters of the puffing and granulating in the step (3) are as follows: the diameter of the screw is 56mm, the length/diameter ratio is 20:1, the screw frequency is 40.6Hz, the feeder is 8.7Hz, the moisture content of the initial material is 28%, the tempering temperature is 90 ℃, the die head temperature is 91 ℃, the steam pressure is 0.3MPa, the steam flow is 11.8kg/h, the water flow rate is 12L/h, the screw rotating speed is 45Hz, the diameter of a die hole is 3mm, the aperture area is 21.21mm 2, the rotating speed of a cutter is 35Hz, the current is 18.8A, the voltage is 380V, and the yield is 200kg/h.
Preferably, the antioxidant is at least one selected from tert-butylhydroquinone (TBHQ), tea polyphenols, propyl gallate and dibutyl hydroxy toluene.
Preferably, the mildew preventive is at least one selected from calcium propionate, fumaric acid, sorbic acid, citric acid and sodium benzoate.
Further preferably, the mildew preventive is specifically calcium propionate and fumaric acid, and the mass ratio of the calcium propionate to the fumaric acid is 1:1.
Preferably, the phagostimulant is at least one selected from glycine, betaine, dimethyl-beta-thiotepa (DMPT) and allicin.
Further preferably, the phagostimulant is specifically glycine and betaine, and the mass ratio of glycine to betaine is 1:2.
Compared with the prior art, the invention has the following beneficial effects: according to the invention, clostridium ethoxide protein, cotton seed concentrated protein, kelp powder containing gelatin and 5-aminolevulinic acid are used as main components of the compound feed for feeding the large yellow croakers, so that the negative influence of fish meal proportion on the health of the large yellow croakers is improved, and meanwhile, the survival rate, the oxidation resistance and the hypoxia stress resistance of the large yellow croakers are improved by combining a specific processing technology and a feed feeding mode. In addition, the nutritional method for improving the anti-hypoxia stress capability of the large yellow croaker has the characteristics of small feed consumption, low production cost and stable effect.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by one of ordinary skill in the art without undue burden from the invention, are within the scope of the invention.
The test methods or test methods described in the following examples are all conventional methods unless otherwise specified; the starting materials and auxiliaries, unless otherwise specified, are obtained commercially from conventional sources or are prepared in conventional manner.
The raw material compositions of examples 1-5 and comparative examples 1-2 are shown in Table 1:
TABLE 1
In Table 1, the amount of 5-ALA added in example 1 was 2mg/kg, the amount of 5-ALA added in example 2 was 6mg/kg, the amount of 5-ALA added in example 3 was 18mg/kg, the amount of 5-ALA added in example 4 was 54mg/kg, and the amount of 5-ALA added in example 5 was 162mg/kg.
In table 1: fish meal a, crude protein, 72.71%, crude fat, 9.46%; clostridium ethoxide protein, crude protein, 83%, crude fat, 5.30%; cottonseed concentrated protein, crude protein, 64.93%, crude fat, 0.92%; baiwei peptide, crude protein, 35.34%, crude fat, 4.61%; standard flour crude protein, 11.50%, crude fat, 1.20%; b mineral premix (mg/kg feed )：FeSO4·H2O,300;ZnSO4·H2O,200;NaCl,100;MnSO4·H2O,25;CuSO4·5H2O,30;CoCl2·6H2O,5;Na2SeO3,5; potassium iodate, 3; zeolite powder, 332.D phagostimulant: glycine: betaine=1:2; e mildewcide: 50% calcium propionate+50% fumaric acid).
A preparation method of a floating compound feed comprises the following specific steps:
(1) Weighing premix, antioxidant (TBQH), mildew inhibitor (calcium propionate, fumaric acid), phagostimulant (glycine, betaine), choline chloride, monocalcium phosphate and kelp powder containing gelatin according to mass ratio, mixing to obtain mixture A, and adding 5-ALA for use;
(2) Pulverizing fish meal, ethanol clostridium protein and cottonseed protein concentrate by a pulverizer, sieving with 178 μm sieve, weighing with Baiwei peptide, tapioca starch, standard flour, ethoxyquinoline and microcrystalline cellulose according to mass ratio, and placing into a mixer for fully mixing for 20min to obtain a mixture B for later use;
(3) Mixing the mixture A and the mixture B, and sequentially puffing, granulating and drying to obtain a semi-finished product of the compound feed;
(4) Uniformly mixing fish oil, soybean oil and soybean lecithin according to a mass ratio to obtain a grease mixture, and then spraying the grease mixture on the surface of a semi-finished product of the compound feed to obtain a finished product of the floating compound feed.
The technological parameters of the puffing and granulating are as follows: the diameter of the screw is 56mm, the length/diameter ratio is 20:1, the screw frequency is 40.6Hz, the feeder is 8.7Hz, the moisture content of the initial material is 28%, the tempering temperature is 90 ℃, the die head temperature is 91 ℃, the steam pressure is 0.3MPa, the steam flow is 11.8kg/h, the water flow rate is 12L/h, the screw rotating speed is 45Hz, the diameter of a die hole is 3mm, the aperture area is 21.21mm 2, the cutter rotating speed is 35Hz, the current is 18.8A, the voltage is 380V, and the yield is 200kg/h;
The process parameters of the drying are as follows: 65 ℃ for 3-4 hours.
Test example 1
The compound feeds prepared in examples 1-5 and comparative examples 1-2 were tested for survival rate, antioxidant capacity, and anti-hypoxia stress capacity, and the experimental procedure was as follows:
a. Feeding process
The cultivation experiment is carried out in a seawater cage cultivation system of a large yellow croaker farm of Fujian Ningde Fufa aquatic products company. Before the formal cultivation experiment, the large yellow croaker is temporarily cultivated in a seawater floating net cage with the size of 4.0x4.0x8.0m to adapt to the environment. After two weeks of starvation for 24 hours, large yellow croakers of a uniform size (initial weight: 17.33.+ -. 0.10 g) were selected and randomly separated into 21 floating cages of 2.0X12.0X12.0 m, each cage 120 tails. Each group of feed was divided into three portions and placed into three net cages, each twice daily satiety feeding (05:00 and 17:30). The water temperature is 19.8-28.5 ℃ during the cultivation period, the salinity is 31.1-34.9, and the dissolved oxygen is more than 6mg/L. The cultivation period is 90 days.
B. Sample collection
After the cultivation experiment is finished, the experimental fish is starved for 24 hours, eugenol diluted 10000 times is used for anesthesia, and the total number of the fish in each net cage is counted and weighed for calculating the survival rate. Taking 10 fish from each net cage, taking blood from tail veins of the fish by using a syringe, standing the collected blood in a refrigerator at 4 ℃ for 4 hours, centrifuging at 4 ℃ for 10 minutes at 3000 revolutions, and collecting serum to be stored in liquid nitrogen. The Malondialdehyde (MDA) content (A003-1-1), total antioxidant capacity (T-AOC), and blood Glucose (GLU) in serum were measured using Nanjing's as-built kit. All experimental procedures for the determination of cortisol hormone (Cortisol) using ELISA kit were carried out exactly according to the instructions.
Where survival (%sr) =100×the end number of experiments/initial mantissa of experiments.
C. Data statistics and analysis
The experimental data were analyzed in single factor by SPSS17.0 software, and the experimental results were expressed as mean ± standard error (x±se), with significant differences (p < 0.05) between the different english alphabets on the upper right hand corner of the same row.
D. Test results
1) The effect of different feeds on the survival rate of large yellow croaker is shown in Table 2.
TABLE 2
As can be seen from Table 2, the survival rate of the large yellow croaker was improved in examples 1 to 5 in which 5-ALA was added at a different concentration gradient, as compared with comparative examples 1 to 2.
2) The effect of different feeds on the antioxidant capacity of large yellow croaker is shown in Table 3.
TABLE 3 Table 3
From Table 3, it can be seen that there is a significant difference (P < 0.05) between comparative example 1 and comparative example 2, and that the T-AOC of examples 1-5 is higher than that of comparative example 1.
The MDA content of examples 2-5 is lower than that of comparative examples 1-2. Meanwhile, the addition of 5-ALA plays an important role in improving the oxidation resistance of the large yellow croaker, and the embodiment 4 can obviously improve the oxidation resistance of the large yellow croaker.
3) The results of the effect of different feeds on the anti-hypoxia stress ability of large yellow croaker are shown in table 4.
TABLE 4 Table 4
As can be seen from Table 4, examples 1-5 provided less blood glucose levels and less cortisol hormone in the body of the large yellow croaker than comparative examples 1-2.
Test example 2
The compound feeds prepared in examples 1-5 and comparative examples 1-2 were subjected to a water stress survival rate test as follows:
a. Experimental procedure
The large yellow croaker water-leaving stress experiment is carried out in a sea water cage culture system of a large yellow croaker culture farm of Fujian Ningde Fu aquatic product company. When the cultivation experiment is finished, firstly, carrying out experiment sampling, leaving 20 fishes in each net cage, placing a plastic water tank with a hole at the bottom in the center of each net cage, when carrying out water-leaving operation, fishing the fishes out of the aquarium with the hole at the bottom for 2min, then placing the fishes into water for 2min, repeating the water-leaving operation for 4 times, then placing the fishes back into the net cage, and sampling after 30 min.
B. Sample collection
After the end of the off-water stress experiment, the total number of surviving fish in each cage was counted for survival rate by anaesthetizing with 10000-fold diluted eugenol.
Where survival (% SR) =100×survival mantissa after water stress/initial mantissa of experiment.
C. Test results
The results of the effect of different feeds on survival rate of large yellow croaker after water stress are shown in Table 5.
TABLE 5
Treatment group Survival rate after stress SR%
Example 1 (30% fish meal+2 mg/kg 5-ALA) 85.00±2.89bc
Example 2 (30% fish meal +6mg/kg 5-ALA) 91.67±1.67abc
Example 3 (30% fish meal +18mg/kg 5-ALA) 95.00±2.89ab
Example 4 (30% fish meal +54mg/kg 5-ALA) 96.67±1.67a
Example 5 (30% fish meal +162mg/kg 5-ALA) 93.33±1.67ab
Comparative example 1 (40% fish meal +0mg/kg 5-ALA) 81.67±1.67c
Comparative example 2 (30% fish meal +0mg/kg 5-ALA) 70.00±2.89d
As can be seen from table 5, the survival rate after water stress was significantly lower for comparative example 2 than for comparative example 1 (P < 0.05); as the addition concentration of 5-ALA increased, the survival rate of large yellow croaker showed a tendency to increase and decrease after water-off stress, examples 3 to 5 had a significant difference (P < 0.05) from comparative example 1, and example 4 had the highest survival rate of large yellow croaker after water-off stress. The survival rate of the large yellow croaker provided in examples 1-5 after water stress is higher than that of comparative examples 1-2.
From the above, the effect of example 4 was most remarkable as a result of analysis of the above test results. After the fish meal is partially replaced by the clostridium ethoxide protein and the cottonseed protein concentrate in the compound feed, a certain proportion of 5-ALA is added, so that the survival rate, the antioxidant capacity and the anti-stress capacity of the large yellow croaker can be improved, the negative influence on the body health of the large yellow croaker caused by the reduction of the fish meal proportion in the aquatic feed is overcome, the use of the fish meal is saved to a certain extent, the production cost is reduced, and the economic benefit is improved.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims (5)

1. A nutritional method for improving the anti-hypoxia stress capability of large yellow croaker, comprising the steps of preparing, processing, feeding and culturing feed, which is characterized by further comprising the following steps:
mixing ethanol clostridium protein, cotton seed concentrated protein, kelp powder containing gelatin and 5-aminolevulinic acid with basic feed, and performing puffing granulation, drying and post-spraying to obtain the large yellow croaker floatability compound feed; the floating compound feed is put into a culture pond of large yellow croaker, and is fed twice a day, and the time interval between the two times is 12-13 hours.
2. The nutritional method for improving the anti-hypoxia stress capacity of large yellow croakers according to claim 1, wherein the addition amount of the 5-aminolevulinic acid in the floating compound feed is 2-162 mg/kg.
3. The nutritional method for improving the anti-hypoxia stress capacity of large yellow croaker according to claim 1, wherein the ingredients of the basal feed comprise: fish meal, herborist peptide, tapioca starch, standard flour, fish oil, soybean lecithin, premix, choline chloride, monocalcium phosphate, mildew inhibitor, antioxidant, ethoxyquinoline, microcrystalline cellulose and phagostimulant.
4. A nutritional method for improving the anti-hypoxia stress ability of large yellow croaker according to claim 3, wherein the preparation method of the floating compound feed comprises the following steps:
(1) 0.5 to 2 weight percent of premix, 0.01 to 0.02 weight percent of antioxidant, 0.05 to 0.1 weight percent of mildew preventive, 0.15 to 0.3 weight percent of phagostimulant, 0.1 to 0.3 weight percent of choline chloride, 0.8 to 2 weight percent of monocalcium phosphate and 0.5 to 1.5 weight percent of kelp powder containing gelatin are weighed and mixed uniformly according to the mass ratio to obtain a mixture A, and 5-aminolevulinic acid is added for standby;
(2) Weighing and uniformly mixing 30-40 wt% of fish meal, 7.14-10.08 wt% of clostridium ethoxide protein, 18.26-25.76 wt% of cottonseed concentrated protein, 4-5 wt% of Baiwei peptide, 4-5 wt% of tapioca starch, 10-12 wt% of standard flour, 0.03-0.06 wt% of ethoxyquinoline and 2.29-3.38wt% of microcrystalline cellulose according to a mass ratio to obtain a mixture B for standby;
(3) Mixing the mixture A and the mixture B, and sequentially puffing, granulating and drying to obtain a semi-finished product of the compound feed;
(4) Uniformly mixing 3.9-6wt% of fish oil, 1-3wt% of soybean oil and 1-1.5wt% of soybean lecithin according to a mass ratio to obtain a grease mixture, and then spraying the grease mixture on the surface of a semi-finished product of the compound feed to obtain a finished product of the compound feed.
5. The nutritional method for improving the anti-hypoxia stress capacity of large yellow croaker according to claim 4, wherein the process parameters of the puffing granulation in the step (3) are as follows: the diameter of the screw is 56mm, the length/diameter ratio is 20:1, the screw frequency is 40.6Hz, the feeder is 8.7Hz, the moisture content of the initial material is 28%, the tempering temperature is 90 ℃, the die head temperature is 91 ℃, the steam pressure is 0.3MPa, the steam flow is 11.8kg/h, the water flow rate is 12L/h, the screw rotating speed is 45Hz, the diameter of a die hole is 3mm, the aperture area is 21.21mm 2, the rotating speed of a cutter is 35Hz, the current is 18.8A, the voltage is 380V, and the yield is 200kg/h.
CN202410358921.9A 2024-03-27 2024-03-27 Nutritional method for improving anti-hypoxia stress capability of large yellow croaker Pending CN118303345A (en)

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