CN115500433A - Method for balancing fatty acid in fish feed, feed and application - Google Patents
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Abstract
The invention relates to a method for balancing fatty acid in fish feed, feed and application, belonging to the field of aquatic product nutritional feed, wherein for fish with muscle polar lipid content of less than 70% of total lipid content, the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5:2.5:1.5:1. Wherein the mass percentages of SFA, MUFA, 18C-PUFA and LC-PUFA in the total fatty acid are respectively 25%, 15% and 10%. For fish with muscle polar lipid content higher than 70% of total lipid content, the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5:3:1.2:1. Wherein the percentage of SFA, MUFA, 18C-PUFA and LC-PUFA in the total fatty acid is 25%, 30%, 12% and 10% respectively. The method of the invention can save 86% of fish oil in the feed and greatly reduce the formula cost on the premise of not influencing growth, survival and muscle quality.
Description
Technical Field
The invention belongs to the field of aquatic product nutritional feeds, and particularly relates to a method for balancing fatty acid in a fish feed, a feed and application.
Background
With the rapid development of aquaculture, the supply of aquatic products in the world has become more and more dependent on the end of aquaculture. The current contribution of aquaculture to consumer water products has exceeded the fishing industry. However, another problem with the rapid development of aquaculture is the short supply of fish meal and fish oil resources as one of the main feed materials. Particularly, in China, the aquaculture yield accounts for more than 70% of the total world yield, and more than half of the fish meal and fish oil resources for feeds in the world are consumed. For fish meal shortage, potential solutions exist, such as the development of plant materials to replace fish meal, and the amino acid composition of the protein contained in fish can be found in terrestrial materials. However, for fish oil shortages, finding suitable alternative oil sources is difficult because long-chain polyunsaturated fatty acids (LC-PUFAs) such as 22. Based on this, an important problem that is put forward by aquaculture practitioners is how to utilize the long-chain polyunsaturated fatty acids in marine fish oil resources to the greatest extent and most efficiently.
It is well known that fat is the most important source of energy for fish in addition to protein, because fish do not make good use of carbohydrates for energy supply. The nature of fat is glycerol and fatty acids, which are the most important material bases of fat as an energy source. The fat content of fish can be divided into three main categories, namely: saturated fatty acids (SFA, mainly including 14, 16, 0, 18, 1 and 20, etc.), monounsaturated fatty acids (MUFA, mainly including 16, 1n-7, 18, 1n-9, 20, 1n-9 and 22. Wherein the polyunsaturated fatty acids are mainly classified into n-3 series and n-6 series. The n-3 series polyunsaturated fatty acids mainly comprise 18; the n-6 series polyunsaturated fatty acids mainly comprise 18 n-6, 20.
The preference of fatty acids of different chain lengths and desaturation as energy supplying substrates is different. Many fish tend to preferentially utilize both Saturated (SFA) and monounsaturated fatty acids (MUFA) as substrates for energy supply (of course, the order of preferential utilization of different specific fatty acid monomers will vary from fish to fish). Furthermore, 18-carbon polyunsaturated fatty acids (18C-PUFAs) such as 18. In the case of a sufficient LC-PUFA supply or a disproportionate ratio of SFA/MUFA/PUFA fractions, LC-PUFAs such as DHA and EPA are oxidatively supplied by the beta-oxidation method. As the LC-PUFA such as DHA and EPA plays an important role in maintaining the health of fishes and human consumers (plays an extremely important physiological role in maintaining the normal functions of systems such as nerves, vision and reproduction), if the excessive LC-PUFA is used for oxidation energy supply, the negative effects on the growth and health of fishes and the health of human consumers are finally caused. Another negative side, since fish oil is currently the most dominant source of LC-PUFA, the use of excess LC-PUFA for energy supply would be a waste of precious fish oil resources.
In order to solve these problems, in the process of blending fatty acids in aquatic feeds, it is desired to ensure that SFA, MUFA and 18C-PUFA (which can be derived from alternative fat sources such as vegetable oils and fats for terrestrial animals) are supplied in an amount as large as possible, and that LC-PUFA, which is mainly derived from fish oil, is supplied in an amount as small as possible, so as to ensure that LC-PUFA, which is mainly derived from fish oil, is mainly used in processes of maintaining important physiological functions such as cell membrane fluidity. This process is also called the development of LC-PUFA sparing effects of SFA, MUFA and 18C-PUFA.
In the development process, the key technical point is to ensure the most reasonable proportion of SFA/MUFA/18C-PUFA/LC-PUFA so as to maximize the LC-PUFA saving effect of SFA, MUFA and 18C-PUFA. However, the progress made in view of the literature published at present is still very limited. Some scattered results cannot provide solid support for systematic technology development, and the applicable range of cultivation varieties is very limited.
Disclosure of Invention
The invention aims to solve the technical problem of matching feed fatty acid composition according to the fat composition (mainly the proportion of polar fat and neutral fat), the fatty acid composition and the fatty acid requirement of target carnivorous cultured fish, realizing the balance of SFA/MUFA/18C-PUFA/LC-PUFA, and achieving the purposes of maximally promoting growth and keeping the health of fish bodies under the condition of saving fish oil. The method is aimed at the muscle fatty acid composition condition of carnivorous mariculture fishes, and through a large amount of data accumulation, a proper SFA/MUFA/18C-PUFA/LC-PUFA balance method is explored, so that the saving effect of SFA, MUFA and 18C-PUFA on LC-PUFA can be stimulated.
The invention is realized by the following technical scheme:
a method for balancing fatty acid in fish feed comprises the following steps:
for fishes with muscle polar lipid content accounting for less than 70% of total lipid content (such as Scophthalmus maximus, paralichthys olivaceus, lateolabrax japonica, epinephelus coioides, sebastes schlegeli, and Larmichthys crocea), the proportion of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5; wherein SFA (including 14, 16, 0, 18, and 20).
For fish (such as Takifugu rubripes, gadus morhuua, nibea albiflora, etc.) with muscle polar lipid content higher than 70% of total lipid content, the feed is controlled to have SFA/MUFA/18C-PUFA/LC-PUFA ratio: 2.5:3:1.2:1. Among them, SFA (mainly including 14, 16, 0, 18, and 20.
The invention also provides a fish feed prepared by the method.
The invention also provides application of the fish feed.
Compared with the prior art, the invention has the beneficial effects that:
(1) In the field, fishes with more than 70% of polar lipid content in total muscle lipid are considered as lean muscle type fishes, and fishes with less than 70% of polar lipid content are considered as general type or lipid-rich type fishes, so the method of the invention sets the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed aiming at fishes with different muscle polar lipid content. The fatty acid with corresponding proportion is added into the fish feed, so that 86 percent of fish oil in the feed can be saved on the premise of not influencing growth, survival and muscle quality, and the formula cost can be greatly reduced under the background of high price enterprises of the fish oil. Furthermore, only a slight decrease in the content of long-chain polyunsaturated fatty acids such as 22 n-3 (DHA), 20 n-3 (EPA) and 20 n-6 (ARA) in muscle (DHA average content 92.8% of fish oil control) was observed, and in pilot scale experiments, even examples were found in which the EPA content of the experimental group exceeded that of the control group using the method of the present invention.
(2) The fat raw materials such as linseed oil, soybean oil, sunflower seed oil, rapeseed oil, palm oil and chicken oil adopted in the fatty acid balance method adopted by the technology are easily available in the feed industry, and the cost is low, so that the method has very strong operability.
(3) Besides economic benefits, the saving of fish oil also has a series of ecological and social benefits. Firstly, the efficient utilization of the fat source of the low-value feed is beneficial to solving the problem that the feed raw material supply in China is internationally traded for neck blocking; and secondly, the reduction of the using amount of the fish oil can reduce the dependence of aquaculture on fishery fishing and fish powder fish oil production in the global range, and is beneficial to the protection of wild fishery resources.
Drawings
Fig. 1 shows the survival rate of the experimental fish in each treatment group of example 1. Data are expressed as mean ± standard error (n = 4); there were significant differences between data columns that did not contain the same letter (P < 0.05).
Figure 2, experimental fish weight gain for each treatment group in example 1. Data are expressed as mean ± sem (n = 4); there were significant differences between data columns that did not contain the same letter (P < 0.05).
Figure 3, experimental fish muscle DHA, EPA and ARA content for each treatment group in example 1. Data are expressed as mean ± standard error (n = 4); there were significant differences between data columns that did not contain the same letter (P < 0.05).
Fig. 4 shows the survival rate of the experimental fish in each treatment group of example 2. Data are expressed as mean ± standard error (n = 4); there were significant differences between data columns that did not contain the same letter (P < 0.05).
Fig. 5, experimental fish weight gain for each treatment group in example 2. Data are expressed as mean ± standard error (n = 4); there were significant differences between data columns that did not contain the same letter (P < 0.05).
Figure 6, experimental fish muscle DHA, EPA and ARA content for each treatment group in example 2. Data are expressed as mean ± standard error (n = 4); there were significant differences between data columns that did not contain the same letter (P < 0.05).
FIG. 7 shows the survival rate of each treatment group in example 3. Data are presented as mean values (no duplicate cement pools).
The weight gain of each treatment group in fig. 8 and example 3. Data are presented as mean ± standard deviation (n =50 samples). Because no repeated cement pool is arranged, mathematical statistics is not carried out.
Figure 9, experimental fish muscle DHA, EPA and ARA content for each treatment group in example 3. Data are expressed as mean ± standard deviation (n =10 samples). Because no repeated cement pool is arranged, mathematical statistics is not carried out.
FIG. 10 shows the survival rate of each treatment group in example 4. Data are presented as mean values (no duplicate boxes).
The weight gain of each treatment group in fig. 11 and example 4. Data are presented as mean ± standard deviation (n =50 samples). Because no repeated net cages are arranged, mathematical statistics is not carried out.
Figure 12, experimental fish muscle DHA, EPA and ARA content for each treatment group in example 4. Data are presented as mean ± standard deviation (n =10 samples). Because no repeated net cages are arranged, mathematical statistics is not carried out.
Detailed Description
The technical features of the present invention are further explained below by way of examples, but the scope of the present invention is not limited in any way by the examples.
Example 1 Effect evaluation test in turbot culture
1. Experimental design and experimental feed formulation (the basic feed formulation is a simulated common commercial feed formulation, and the invention is not limited in protection scope, and the effect of the invention can be achieved by implementing the nutriology method of the invention under the condition of ensuring the normal growth of the cultured fish)
A feed fatty acid balancing technology and its application in seawater fish, its method is as follows:
for fish with muscle polar lipid content less than 70% of total lipid content (the experimental animal in this experiment is turbot Scophthalmus maximus), the ratio of Saturated Fatty Acid (SFA)/monounsaturated fatty acid (MUFA)/18-carbon polyunsaturated fatty acid (18C-PUFA)/long-chain polyunsaturated fatty acid (LC-PUFA) in the feed is controlled as follows: 2.5:2.5:1.5:1. Among these, SFA (mainly including 14, 16, 0, 18, and 20.
The experimental feed takes fish meal, soybean protein concentrate, soybean meal, millet flour and other raw materials as main protein sources. The crude fat content was about 10-11%, and the 4 groups of experimental feeds used different fat source matching programs to obtain different fatty acid compositions (with SFA/MUFA/18C-PUFA/LC-PUFA ratio as core index) and fish oil as control group (table 1). The feeds of each group were designated as A (fish oil control group), B (fatty acid balanced group), C (high SFA group), D (high MUFA group) and E (high 18C-PUFA group), respectively. The results of the fatty acid testing for each feed group are shown in table 2.
Table 1 feed formulation and coarse content (% dry matter) of experimental feeds
Table 2 experiment feed fatty acid composition (% total fatty acids)
SFA: a saturated fatty acid; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Fish and culture management for experiment
The initial weight of the juvenile turbot is 9g, and before formal test, the experimental fish is temporarily cultured in a cement pond with a square meter of 25 for 2 weeks so as to adapt to the experimental environmental conditions. Before the start of the official experiment, the experimental fish were randomly divided into 20 polyethylene buckets (height: 100cm; diameter: 230 cm), and each group was repeated 4 times, 45 fish per bucket. The cultivation process adopts indoor running water cultivation, and the cultivation seawater adopts underground deep well water. Feeding for 2 times each day. The culture experiment is carried out in yellow sea aquaculture limited company (flounder breeding base) in Haiyang city of Chinese aquatic science institute of yellow sea aquaculture, and the culture period is 8 weeks. The cultivation experiments were carried out under natural photoperiod and ambient temperature (haiyang city, shandong province, china, N36 DEG 41', E121 DEG 07'). In the feeding and breeding experiment process, the water temperature range is 14-18 ℃; salinity of 29 to 31; pH of 7.2-8.4; 6-8 mg L of dissolved oxygen -1 . And (4) removing residual feed and excrement by a siphon method half an hour after the end of daily ingestion.
3. Terminal weight measurement, sample collection and fatty acid analysis
After the breeding experiment is finished, the total weight of each barrel of fish is weighed, and the total number is counted to calculate the survival rate. Meanwhile, 5 fish are dissected, the weight of the liver and internal organs is weighed, and the body length is measured for calculating the liver body ratio, the internal organ ratio and the fullness. The method for calculating the liver body ratio, the viscera body ratio and the fullness comprises the following steps: liver body ratio (%) = liver weight (fresh weight)/body weight × 100; visceral volume ratio (%) = internal organsWeight (fresh weight)/body weight × 100; fullness = body weight (g)/length (cm) 3 . In addition, a muscle (dorsal muscle) sample of 6-tailed fish was taken, and the fatty acid content was measured as follows: the lyophilized samples were esterified with KOH-methanol and HCl-methanol, respectively (72 ℃ water bath), and the methyl-esterified fatty acids were extracted with n-hexane and then measured on the machine. The gas chromatography was carried out using shimadzu GC-2010Pro (japan), a quartz capillary chromatography column (SHRT-2560, 100mx 0. The column temperature was programmed from 150 to 200 ℃ at 15 ℃ per minute and then from 200 to 250 ℃ at 2 ℃ per minute. The injection port and detector temperatures were both 250 ℃. The fatty acids are expressed as a ratio of a certain fatty acid to the total fatty acids.
4. Experimental statistical method
Statistical one-way analysis of variance of experimental data was performed using SPSS16.0 and pairwise comparison analysis was performed using Tukey's method. Data are expressed as mean ± sem (n = 4). The difference is marked by P < 0.5.
5. Results of the experiment
The experiment mainly evaluates the application effect of the technology of the invention by the survival rate, the weight gain rate and the composition of muscle fatty acid.
In the experimental process, the final survival rate has no significant difference and is over 90 percent (figure 1). The fatty acid balanced group (group B) of the invention has no significant difference in weight gain rate from the fish oil control group (group A) (P > 0.05, FIG. 2). The weight gain of the high 18C-PUFA group (group E) is significantly lower than that of the fatty acid balance group (group B) and the fish oil control group (group A) (P < 0.05), and has no significant difference with the high MUFA group (group D). The high SFA group (group C) had the lowest weight gain, significantly lower than the other groups (P < 0.05).
Similar fatty acid compositions were obtained in groups a and B with respect to muscle fatty acid composition (table 3). In particular, in the case of long-chain polyunsaturated fatty acids, only slightly lower contents of ARA, EPA and DHA were obtained in group B than in the fish oil control group (group A). It is shown that the fatty acid composition of group B is very balanced, able to maximally retain long-chain polyunsaturated fatty acids such as ARA, EPA and DHA for muscle accumulation. While the ARA, EPA and DHA content of the muscles of the C, D and E groups were reduced to different extents. B. C, D and group E muscles have DHA contents of 94.3%, 59.7%, 52.3% and 49.1% of group A, respectively (FIG. 3); B. c, D and group E muscle EPA content 90.7%, 22.7%, 35.8% and 51.4% of group a, respectively (fig. 3); B. c, D and E muscle ARA content were 97.2%, 93.2%, 86.4% and 90.4% of group A, respectively (FIG. 3).
Table 3 turbot muscle fatty acid composition (% total fatty acids, mean ± sem, n = 4)
There were significant differences between data columns in the same row that did not contain the same letter (P < 0.05). SFA: a saturated fatty acid; MUFA: mono-unsaturated fatty acids; PUFA: polyunsaturated fatty acids.
Example 2 Effect evaluation test of Using the method in Fugu rubripes cultivation
1. Experimental design and experimental feed formulation (the basic feed formulation is a simulated common commercial feed formulation, and is not limited to the protection scope of the invention, and the effect of the invention can be achieved by implementing the nutrition method of the invention under the condition that the farmed fish can grow)
A feed fatty acid balancing technology and its application in seawater fish, its method is as follows:
for fish with muscle polar lipid content higher than 70% of total lipid content (Takifugu rubripes in this example), the feed ratio of SFA/MUFA/18C-PUFA/LC-PUFA is controlled as follows: 2.5:3:1.2:1. Among these, SFA (mainly including 14, 16, 0, 18, and 20.
The experimental feed takes fish meal, soybean protein concentrate, soybean meal, millet flour and other raw materials as main protein sources. The crude fat content was about 10-11%, and the 4 groups of experimental feeds used different fat source matching programs to obtain different fatty acid compositions (with SFA/MUFA/18C-PUFA/LC-PUFA ratios as the core index) and fish oil as the control group (table 4). The feeds of each group were designated as A (fish oil control group), B (fatty acid balanced group), C (high SFA group), D (high MUFA group) and E (high 18C-PUFA group), respectively. The results of the fatty acid testing for each feed group are shown in table 5.
Table 4 feed formulation and coarse content (% dry matter) of experimental feeds
Table 5 experiment feed fatty acid composition (% total fatty acids)
SFA: a saturated fatty acid; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Fish and culture management for experiment
The test adopts the juvenile fish of the takifugu rubripes with the initial weight of 12g, and before formal test, the test fish is temporarily cultured in a cement pond with the weight of 25 square meters for 10 days so as to adapt to the test environmental conditions. Before the start of the official experiment, the experimental fish were randomly divided into 20 polyethylene buckets (height: 100cm; diameter: 230 cm), and each group was repeated 4 times, with 35 fish per bucket. The cultivation process adopts indoor running water cultivation, and the cultivation seawater adopts underground deep well water. The artificial feeding is carried out for 2 times per day. The culture experiment is carried out in a limited yellow sea aquatic product company (flounder breeding base) in Haiyang city of China institute for aquatic product science, yellow sea, and the culture period is 8 weeks. The cultivation experiment is thatNatural photoperiod and ambient temperature (Shandong province, shanyang city, china, N36 DEG 41', E121 DEG 07'). In the feeding and breeding experiment process, the water temperature range is 14-18 ℃; salinity of 29 to 31; pH of 7.2-8.4; 6-8 mg L of dissolved oxygen -1 . After the ingestion of the feed each day, the residual feed and excrement are removed by a siphon method in half an hour.
3. Terminal weight measurement, sample collection and fatty acid analysis
After the culture experiment is finished, the total weight of each barrel of fish is weighed, and the total number is counted to calculate the survival rate. Meanwhile, 5 fish are dissected, the weight of the liver and internal organs is weighed, and the body length is measured for calculating the liver body ratio, the internal organ ratio and the fullness. The method for calculating the liver body ratio, the viscera body ratio and the fullness comprises the following steps: liver body ratio (%) = liver weight (fresh weight)/body weight × 100; visceral volume ratio (%) = visceral weight (fresh weight)/body weight × 100; fullness = body weight (g)/length (cm) 3 . In addition, a muscle (dorsal muscle) sample of 6-tailed fish was taken, and the fatty acid content was measured as follows: the lyophilized samples were esterified with KOH-methanol and HCl-methanol, respectively (72 ℃ water bath), and the methyl-esterified fatty acids were extracted with n-hexane and then measured on the machine. The gas chromatography was carried out using shimadzu GC-2010Pro (japan), a quartz capillary chromatography column (SHRT-2560, 100mx 0. The column temperature was programmed from 150 to 200 ℃ at 15 ℃ per minute and then from 200 to 250 ℃ at 2 ℃ per minute. The injection port and detector temperatures were both 250 ℃. Fatty acids are expressed as a ratio of certain fatty acids to the total fatty acids.
4. Experimental statistical method
Statistical one-way analysis of variance of experimental data was performed using SPSS16.0 and pairwise comparative analysis was performed using Tukey's method. Data are expressed as mean ± sem (n = 4). The difference is marked by P < 0.5.
5. Results of the experiment
The experiment mainly evaluates the application effect of the technology according to the survival rate, the weight gain rate and the composition of the muscle fatty acid.
In the experimental process, the final survival rate has no significant difference and is over 84 percent (figure 4). The fatty acid-balanced group (group B) according to the present invention showed no significant difference in weight gain from the fish oil control group (group a) (P > 0.05, fig. 5). The weight gain of the high MUFA group (group D) and the high 18C-PUFA group (group E) is significantly lower than that of the fatty acid balance group (group B) and the fish oil control group (group A) (P < 0.05). The high SFA group (group C) was not significantly different from each of the other groups.
Very similar to the results on turbot, similar fatty acid compositions were obtained in groups a and B in terms of muscle fatty acid composition (table 6). Especially in the case of long chain polyunsaturated fatty acids, group B only achieved slightly lower ARA, EPA and DHA content than fish oil control group (group a), indicating that the fatty acid composition of group B is very balanced, and long chain polyunsaturated fatty acids such as ARA, EPA and DHA can be maximally retained for muscle accumulation. Especially DHA and EPA content, there was no significant difference between group a and group B. While the muscle ARA, EPA and DHA content of the C, D and E groups were reduced to different extents. B. C, D and group E muscles have DHA contents of 90.6%, 69.3%, 60.9% and 64.6% of group a, respectively (fig. 6); B. c, D and group E muscle EPA contents were 86.7%, 55.5%, 70.3% and 43.3% of group a, respectively (fig. 6); B. c, D and group E muscle ARA contents were 75.8%, 66.7% and 69.7% of group A, respectively (FIG. 6).
Table 6 experiment of fugu rubripes muscle fatty acid composition (% total fatty acids, mean ± sem, n = 4)
There were significant differences between data columns in the same row that did not contain the same letter (P < 0.05). SFA: a saturated fatty acid; MUFA: mono-unsaturated fatty acids; PUFA: a polyunsaturated fatty acid.
Example 3 Effect evaluation of the application of the method in a Pilot-scale experiment of turbot
1. Experimental design and experimental feed formulation (the basic feed formulation is a simulated common commercial feed formulation, and is not limited to the protection scope of the invention, and the effect of the invention can be achieved by implementing the nutrition method of the invention under the condition that the farmed fish can grow)
A feed fatty acid balancing technology and its application in seawater fish, its method is as follows:
for fishes with muscle polar lipid content less than 70% of total lipid content (the experimental animal in the experiment is Scophthalmus maximus), the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5:2.5:1.5:1. Among these, SFA (mainly including 14, 16, 0, 18, and 20.
Due to pilot scale experimental site limitations, only group a and group B of the examples were used as experimental feeds in this example. The experimental feed formulation and manufacturing method were the same as in example 1. Namely, the experimental feed takes fish meal, soybean protein concentrate, soybean meal, millet flour and other raw materials as main protein sources. Crude fat content was about 10-11%, different fatty acid compositions were obtained using different fat source matching programs for the 2 experimental feeds, fish oil group as control group (group a), and fatty acid balance group using the method as experimental group (group B) (table 7). The results of the fatty acid testing of the two experimental feeds are shown in table 8.
TABLE 7 feed formulation and coarse content (% dry matter) of experimental feeds
Table 8 experimental feed fatty acid composition (% total fatty acids)
SFA: a saturated fatty acid; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Fish and farming management for experiments
The experiment adopts the experimental fish with the initial weight of 152g, 2 groups of experimental feeds are respectively fed into 2 indoor seawater culture cement ponds (3 m multiplied by 1.5 m), 300 experimental fishes are placed in each cement pond, the culture experiment is carried out in yellow sea aquaculture limited company in Haiyang city in cigarette platform city, the culture water is the seawater of underground deep wells, and running water culture is adopted. Feeding twice a day with a total cultivation period of 10 weeks (70 days).
3. Terminal body weight measurement, sample collection and fatty acid analysis
And after the culture experiment is finished, randomly sampling 50 tail fishes in each cement pond, weighing and calculating the weight gain rate. Meanwhile, 10 muscle (dorsal muscle) samples of the fish were taken randomly per cage, and the fatty acid content was determined as follows: the lyophilized samples were esterified with KOH-methanol and HCl-methanol, respectively (72 ℃ water bath), and then methyl-esterified fatty acids were extracted with n-hexane and then measured on a machine. Gas chromatography was performed using shimadzu GC-2010Pro (japan), a quartz capillary column (SHRT-2560, 100mx 0 × 25mm × 0. The column temperature was programmed from 150 ℃ to 200 ℃ at 15 ℃ per minute and then from 200 ℃ to 250 ℃ at 2 ℃ per minute. The injection port and detector temperatures were both 250 ℃. Fatty acids are expressed as a ratio of a certain fatty acid to the total fatty acids.
4. Experimental statistical method
Since no duplicate cement pools were designed, only the mean and standard deviation were calculated and no statistical analysis was done.
5. Results of the experiment
In terms of survival rate, the survival rates of group a and group B were 94.7% and 93%, respectively, with no significant difference (fig. 7). Group B (163%) was even slightly higher than group a (150%) in terms of weight gain (fig. 8). In terms of muscle long chain polyunsaturated fatty acid content (fig. 9), DHA, EPA and ARA contents in group B were 96%, 74.6% and 169%, respectively, of group a. The ARA content in group B even exceeds that in group A.
Example 4 Effect evaluation of Using the method in Fugu rubripes Pilot-Scale experiments
1. Experimental design and experimental feed formulation (the basic feed formulation is a simulated common commercial feed formulation, and is not limited to the protection scope of the invention, and the effect of the invention can be achieved by implementing the nutrition method of the invention under the condition that the farmed fish can grow)
A feed fatty acid balancing technology and its application in seawater fish, its method is as follows:
for fish with muscle polar lipid content higher than 70% of total lipid content (Takifugu rubripes in this example), the feed ratio of SFA/MUFA/18C-PUFA/LC-PUFA is controlled as follows: 2.5:3:1.2:1. Among these, SFA (mainly including 14, 16, 0, 18, and 20.
The experiment used the same feed formulation as in example 2, and only two treatment groups A, B were used due to pilot plant scale limitations. Namely, the experimental feed takes fish meal, soybean protein concentrate, soybean meal, millet flour and other raw materials as main protein sources. Crude fat content was about 10-11%, different fatty acid compositions were obtained using different fat source matching programs for the 2 groups of experimental feeds (SFA/MUFA/18C-PUFA/LC-PUFA ratio as core index), fish oil group as control group (group a), and fatty acid balanced feed group using the method of the invention as treatment group (group B) (table 9). The feed fatty acid composition is shown in table 10.
Table 9 feed formulation and coarse content (% dry matter) of experimental feeds
Table 10 experimental feed fatty acid composition (% total fatty acids)
SFA: a saturated fatty acid; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Fish and culture management for experiment
The experiment adopts experimental fish with initial weight of 109g, 2 groups of experimental feeds are respectively fed to 2 mariculture net cages (3 m multiplied by 2 m), 400 pieces of experimental fish are placed in each net cage pond, and the culture experiment is carried out in Tangshan City in Hebei river. Feeding twice a day with a total cultivation period of 12 weeks (84 days).
3. Terminal body weight measurement, sample collection and fatty acid analysis
And after the culture experiment is finished, randomly sampling 50 tail fishes in each cement pond, weighing and calculating the weight gain rate. Meanwhile, 10 muscle (dorsal muscle) samples of the fish were taken randomly per cage, and the fatty acid content was determined as follows: the lyophilized samples were esterified with KOH-methanol and HCl-methanol, respectively (72 ℃ water bath), and then methyl-esterified fatty acids were extracted with n-hexane and then measured on a machine. The gas chromatography was carried out using Shimadzu GC-2010Pro (Japan), a quartz capillary column (SHRT-2560, 100mx 0. The column temperature was programmed from 150 ℃ to 200 ℃ at 15 ℃ per minute and then from 200 ℃ to 250 ℃ at 2 ℃ per minute. The injection port and detector temperatures were both 250 ℃. Fatty acids are expressed as a ratio of a certain fatty acid to the total fatty acids.
4. Experimental statistical method
Because no duplicate cages were designed, only the mean and standard deviation were calculated and no statistical analysis was done.
5. Results of the experiment
In terms of survival rate, the survival rates of the group A and the group B were 81.3% and 85.5%, respectively, with no significant difference, and the survival rate of the group B was even slightly higher than that of the group A (FIG. 10). Group B (368%) was also slightly higher in weight gain than group a (324%) (fig. 11). In terms of muscle long chain polyunsaturated fatty acid content (fig. 12), DHA, EPA and ARA contents in group B were 90.2%, 115.6% and 106.3% of group a, respectively. The EPA and ARA content of the group B exceeds that of the group A.
Claims (3)
1. A method for balancing fatty acid in fish feed is characterized by comprising the following steps:
(1) For fish with muscle polar lipid content of less than 70% of total lipid content, the ratio of saturated fatty acid/monounsaturated fatty acid/18-carbon polyunsaturated fatty acid/long-chain polyunsaturated fatty acid in the feed is controlled as follows: 2.5; wherein the mass percentages of the saturated fatty acid, the monounsaturated fatty acid, the 18-carbon polyunsaturated fatty acid and the long-chain polyunsaturated fatty acid in the total fatty acid are respectively 25%, 15% and 10%;
(2) For fish with muscle polar lipid content higher than 70% of total lipid content, the ratio of saturated fatty acid/monounsaturated fatty acid/18-carbon polyunsaturated fatty acid/long-chain polyunsaturated fatty acid in the feed is controlled as follows: 2.5; wherein the mass percentages of the saturated fatty acid, the monounsaturated fatty acid, the 18-carbon polyunsaturated fatty acid and the 18-carbon polyunsaturated fatty acid in the total fatty acid are respectively 25%, 30%, 12% and 10%.
2. A fish feed, wherein the feed is formulated according to the method of claim 1.
3. Use of a fish feed according to claim 2.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20090299083A1 (en) * | 2005-11-18 | 2009-12-03 | Matthew Robert Miller | Feedstuffs for Aquaculture Comprising Stearidonic Acid |
| CN107258622A (en) * | 2017-07-12 | 2017-10-20 | 中国水产科学研究院黄海水产研究所 | A kind of dietary methods of regulation and control Cynoglossus semilaevis parent population sex hormone secretion |
| US20190373915A1 (en) * | 2014-10-24 | 2019-12-12 | Trustees Of Dartmouth College | Aquaculture Feed Formulation and Aquaculture Product Produced with the Same |
| CN114847418A (en) * | 2022-05-24 | 2022-08-05 | 华南农业大学 | Composite fat source for improving muscle quality of tilapia based on accurate nutritional requirement of fatty acid |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090299083A1 (en) * | 2005-11-18 | 2009-12-03 | Matthew Robert Miller | Feedstuffs for Aquaculture Comprising Stearidonic Acid |
| US20190373915A1 (en) * | 2014-10-24 | 2019-12-12 | Trustees Of Dartmouth College | Aquaculture Feed Formulation and Aquaculture Product Produced with the Same |
| CN107258622A (en) * | 2017-07-12 | 2017-10-20 | 中国水产科学研究院黄海水产研究所 | A kind of dietary methods of regulation and control Cynoglossus semilaevis parent population sex hormone secretion |
| CN114847418A (en) * | 2022-05-24 | 2022-08-05 | 华南农业大学 | Composite fat source for improving muscle quality of tilapia based on accurate nutritional requirement of fatty acid |
Non-Patent Citations (1)
| Title |
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| （美）克雷曼原: "全国高职高专食品类、保健品开发与管理专业"十三五"规划教材 功能性食品开发与应用", 中国医药科技出版社, pages: 282 * |
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