CN115500433B - Method for balancing fatty acid in fish feed, feed and application - Google Patents
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Insects & Arthropods (AREA)
- Marine Sciences & Fisheries (AREA)
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- Fodder In General (AREA)
Abstract
The invention relates to a fatty acid balancing method in fish feed, feed and application, which belong to the field of aquatic nutrition feed, and for fish with muscle polar lipid content accounting for 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 SFA, MUFA, 18C-PUFA and LC-PUFA account for 25%, 15% and 10% of the total fatty acids by mass, respectively. 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 control feed is as follows: 2.5:3:1.2:1. Wherein the percentage of SFA, MUFA, 18C-PUFA and LC-PUFA is 25%, 30%, 12% and 10% of total fatty acids, respectively. The method can save 86% of fish oil in the feed on the premise of not affecting growth, survival and muscle quality, and greatly reduces the formula cost.
Description
Technical Field
The invention belongs to the field of aquatic nutrition feed, and particularly relates to a method for balancing fatty acid in fish feed, the feed and application.
Background
With the rapid development of aquaculture, the supply of world-wide aquatic products has become increasingly dependent on the end of aquaculture. The current contribution of aquaculture to consumer products has exceeded the fishing industry. However, another problem with the rapid development of aquaculture is the supply and supply of fish meal fish oil resources as one of the main feed materials. In particular, the aquaculture yield in China is over 70% of the total world yield, and more than half of the world fish meal and fish oil resources for feed are consumed. There are potential solutions for fish meal shortage, such as the development of plant material to replace fish meal, after all the amino acid composition of proteins contained in fish is essentially completely found in terrestrial materials. However, it is difficult to find suitable alternative oil sources for fish oil shortage, because long chain polyunsaturated fatty acids (LC-PUFAs) that are very important for fish growth and health, such as 22:6n-3 (docosahexaenoic acid, DHA), 20:5n-3 (eicosapentaenoic acid, EPA), and 20:4n-6 (arachidonic acid, ARA), contained in fish oils, are hardly present in terrestrial raw materials. Based on this, an important question posed to aquaculture practitioners is how to make the most efficient use of long chain polyunsaturated fatty acids in marine fish oil resources.
It is well known that fat is the most important source of energy for fish other than protein, because fish do not make good use of carbohydrates to supply energy. The nature of fat is glycerol and fatty acids, which are the most important substance basis for fat as an energy source. Fat contained in fish can be mainly divided into three categories, namely: saturated fatty acids (SFA, mainly comprising 14:0, 16:0, 18:1 and 20:0 etc.), monounsaturated fatty acids (MUFA, mainly comprising 16:1n-7, 18:1n-9, 20:1n-9 and 22:1n-11 etc.), and polyunsaturated fatty acids (PUFA, number of double bonds more than 1, carbon chain length greater than or equal to 18). Wherein polyunsaturated fatty acids are again largely divided into n-3 series and n-6 series. The n-3 series polyunsaturated fatty acids mainly comprise 18:3n-3, 18:4n-3, 20:3n-3, EPA, 22:5n-3, DHA, etc.; the n-6 series polyunsaturated fatty acids mainly comprise 18:2n-6, 20:2n-6, ARA, etc.
Fatty acids of different chain lengths and desaturations differ in priority as energy supply substrates. Many fish tend to preferentially utilize Saturated (SFA) and monounsaturated fatty acids (MUFA) as substrates for energy supply (of course, the order of preference for different specific fatty acid monomers may vary from fish to fish). In addition, 18 carbon polyunsaturated fatty acids (18C-PUFAs) such as 18:2n-6 and 18:3n-3 are also important and efficient sources of energy for fish. In cases where the LC-PUFA supply is sufficient or the ratio of the SFA/MUFA/PUFA fractions is not sufficient, the LC-PUFA such as DHA and EPA will also be oxidatively energized by beta oxidation. Since LC-PUFAs such as DHA and EPA have an important role in maintaining the health of both fish and human consumers (which plays an extremely important physiological role in maintaining the normal functions of the nervous, visual and reproductive systems), the use of excessive LC-PUFAs for oxidative energy can negatively impact fish growth and health and ultimately human consumer health. Another negative, since fish oils are currently the most dominant source of LC-PUFAs, the use of excess LC-PUFAs for energy supply would be wasteful of precious fish oil resources.
In view of these problems, in the process of preparing fatty acids for aquatic feeds, the aim is to ensure that as much SFA, MUFA and 18C-PUFA as possible (which can be derived from vegetable oil, terrestrial animal fat and other alternative fat sources) are used for energy supply, and as little LC-PUFA as possible is used for energy supply, so that the LC-PUFA mainly derived from fish oil is mainly used for important physiological functions such as maintaining cell membrane fluidity and the like. This process is also known as exploiting the LC-PUFA sparing effects of SFA, MUFA and 18C-PUFA.
In this development process, the core technical point is to ensure the most reasonable ratio of SFA/MUFA/18C-PUFA/LC-PUFA to maximize the LC-PUFA-saving effect of SFA, MUFA and 18C-PUFA. However, the progress made is still very limited from the literature published today. Some scattered results cannot provide solid support for systematic technical development, and the applicable cultivation variety range is very limited.
Disclosure of Invention
The invention aims to solve the technical problem that the feed fatty acid composition is proportioned according to the fat composition (mainly the proportion of polar fat and neutral fat) of the target carnivorous farmed fish, the fatty acid composition and the fatty acid requirement, so as to realize the balance of SFA/MUFA/18C-PUFA/LC-PUFA, thereby achieving the aims of promoting the growth to the greatest extent and keeping the health of fish under the condition of saving fish oil. According to the method, a proper SFA/MUFA/18C-PUFA/LC-PUFA balance method is explored through accumulation of a large amount of data according to the muscle fatty acid composition condition of the carnivorous marine cultured fish, and 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 fish with muscle polar lipid content lower than 70% of total lipid content (such as turbot Scophthalmus maximus, paralichthys olivaceus Paralichthys olivaceus, lateolabrax japonicus Lateolabrax japonicus, epinephelus malabaricus Epinephelus coioides, secoitus schlegeli Sebastes schlegeli, and large yellow croaker Larmichthys crocea), the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5:2.5:1.5:1; wherein SFA (including 14:0, 16:0, 18:0 and 20:0), MUFA (including 16:1n-7, 18:1n-9, 20:1n-9 and 22:1 n-11), 18C-PUFA (including 18:2n-6 and 18:3 n-3) and LC-PUFA (including 20:2n-6, 20:3n-3, ARA, EPA, DPA and DHA) account for 25%, 15% and 10% of the total fatty acids, respectively.
For fish with muscle polar lipid content higher than 70% of total lipid content (such as Fugu rubripes Takifugu rubripes, gadus morhua in Atlantic, nibea cobor in Nibea Nib), the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5:3:1.2:1. Wherein SFA (mainly comprising 14:0, 16:0, 18:0 and 20:0), MUFA (mainly comprising 16:1n-7, 18:1n-9, 20:1n-9 and 22:1 n-11), 18C-PUFA (mainly comprising 18:2n-6 and 18:3 n-3) and LC-PUFA (mainly comprising 20:2n-6, 20:3n-3, ARA, EPA, DPA and DHA) account for 25%, 30%, 12% and 10% of the total fatty acids, respectively.
The invention also provides 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, fish with more than 70% of the total muscle lipid content is regarded as lean muscle fish, and fish with less than 70% are regarded as general type or fat-rich fish, so that the method of the invention prepares the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in feed aiming at fish with different muscle polar lipid contents. The fish feed is added with fatty acid in a corresponding proportion, 86% of fish oil in the feed can be saved on the premise of not affecting growth, survival and muscle quality, and the formula cost can be greatly reduced on the background of high price of fish oil. Moreover, only a slight decrease in the content of 22:6n-3 (DHA), 20:5n-3 (EPA) and 20:4n-6 (ARA) long chain polyunsaturated fatty acids in the muscle (average DHA content is 92.8% of that of the fish oil control group) was observed in pilot scale experiments, even in the examples where the EPA content of the experimental group exceeded that of the control group by applying the method of the present invention.
(2) The fatty acid balance method adopted by the technology adopts fat raw materials such as linseed oil, soybean oil, sunflower seed oil, rapeseed oil, palm oil and chicken oil which are all easily available raw materials in the feed industry, and has low cost, so the method has very strong operability.
(3) Besides economic benefits, the fish oil saving also has a series of ecological and social benefits. Firstly, the high-efficiency utilization of the low-value feed fat source is helpful to solve the problem that the feed raw material supply of China is internationally trade' blocked; secondly, the reduction of the use amount of the fish oil reduces the dependence of aquaculture on fishery fishing and fish meal fish oil production in the global scope, and is beneficial to the protection of wild fishery resources.
Drawings
FIG. 1, example 1, each treatment group tested fish survival rate. Data are expressed as mean ± standard error (n=4); there was a significant difference between data columns that did not contain the same letter (P < 0.05).
Fig. 2, example 1, each treatment group tested the weight gain rate of fish. Data are expressed as mean ± standard error (n=4); there was a significant difference between data columns that did not contain the same letter (P < 0.05).
Fig. 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 was a significant difference between data columns that did not contain the same letter (P < 0.05).
FIG. 4 shows the survival rate of fish in each treatment group in example 2. Data are expressed as mean ± standard error (n=4); there was a significant difference between data columns that did not contain the same letter (P < 0.05).
Fig. 5, example 2, each treatment group tested the weight gain rate of fish. Data are expressed as mean ± standard error (n=4); there was a significant difference between data columns that did not contain the same letter (P < 0.05).
Fig. 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 was a significant difference 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 average (no duplicate cement reservoirs were set).
Weight gain ratio for each treatment group in fig. 8 and example 3. Data are expressed as mean ± standard deviation (n=50 samples). Because no repeated cement pool is arranged, no mathematical statistics is carried out.
Fig. 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, no mathematical statistics is carried out.
FIG. 10 shows the survival rate of each treatment group in example 4. Data are presented as average values (no duplicate cage).
Fig. 11, weight gain ratio for each treatment group in example 4. Data are expressed as mean ± standard deviation (n=50 samples). Because no repeated net cage is arranged, no mathematical statistics is performed.
Fig. 12, experimental fish muscle DHA, EPA and ARA content for each treatment group in example 4. Data are expressed as mean ± standard deviation (n=10 samples). Because no repeated net cage is arranged, no mathematical statistics is performed.
Detailed Description
The technical features of the present invention will be further explained by examples below, but the scope of the present invention is not limited in any way by the examples.
Example 1 Effect evaluation test of applying the method in turbot culture
1. Experiment design and experiment feed formula (the basic feed formula is a commercial feed formula which is used for simulating, but is not limited to the protection scope of the invention), and under the condition that the normal growth of the cultured fish can be ensured, the effect of the invention can be achieved by implementing the nutrition method of the invention
A feed fatty acid balancing technology and its application in sea fish, its method is as follows:
for fish with muscle polar lipid content lower 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. Wherein SFA (mainly comprising 14:0, 16:0, 18:0 and 20:0), MUFA (mainly comprising 16:1n-7, 18:1n-9, 20:1n-9 and 22:1 n-11), 18C-PUFA (mainly comprising 18:2n-6 and 18:3 n-3) and LC-PUFA (mainly comprising 20:2n-6, 20:3n-3, ARA, EPA, DPA and DHA) account for 25%, 15% and 10% of total fatty acids, respectively.
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 collocation schemes to obtain different fatty acid compositions (with SFA/MUFA/18C-PUFA/LC-PUFA ratio as core index) and the fish oil group as control group (Table 1). The feeds for each group were designated A (fish oil control group), B (fatty acid balance group), C (high SFA group), D (high MUFA group) and E (high 18C-PUFA group), respectively. The fatty acid test results for each feed group are shown in table 2.
Table 1 feed formulation and crude ingredients (% dry matter) of experimental feed
Table 2 Experimental feed fatty acid composition (% Total fatty acids)
SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Experimental fish and aquaculture management
The experiment adopts an initial bodyJuvenile turbot weighing 9g were temporarily cultured in a cement pond of 25 square meters for 2 weeks to accommodate experimental environmental conditions prior to formal testing. Before the beginning of the formal experiment, the experimental fish were randomly divided into 20 polyethylene barrels (height: 100cm; diameter: 230 cm), each group was repeated 4 times, 45 fish per barrel. Indoor running water culture is adopted in the culture process, and underground deep well water is adopted as the seawater for culture. Fed by manual satiety for 2 times each day. The culture experiments were carried out at the Shaoyang yellow sea aquatic products Limited company (flounder breeding base) of yellow sea aquatic products institute of China aquatic science, and the culture period was 8 weeks. The cultivation experiments were performed at natural photoperiod and ambient temperature (N36°41', E121°07') in the sea-yang city of shandong province, china. In the feeding and breeding experiment process, the water temperature range is 14-18 ℃; salinity, 29-31; the pH is 7.2-8.4; dissolved oxygen, 6-8 mg L -1 . Residual feed and faeces were removed by siphoning half an hour after the end of daily feeding.
3. Terminal body weight measurement, sample collection and fatty acid analysis
After the cultivation experiment is finished, the total weight of each barrel of fish is weighed, and the total weight is counted to calculate the survival rate. Meanwhile, 5 fish are dissected, the weights of livers and viscera are weighed, and the body length is measured and used for calculating the liver-body ratio, the viscera-body ratio and the fullness. The calculation method of the liver body ratio, the viscera body ratio and the fullness comprises the following steps: liver ratio (%) = liver weight (fresh weight)/body weight×100; visceral volume ratio (%) = visceral weight (fresh weight)/body weight x 100; fullness = body weight (g)/body length (cm) 3 . A muscle (dorsum muscle) sample of 6 fish was taken and the fatty acid content was determined as follows: the lyophilized samples were esterified with KOH-methanol and HCl-methanol, respectively (72 ℃ C. Water bath), and the methyl-esterified fatty acids were extracted with n-hexane and then measured on an engine. Gas chromatography was carried out using Shimadzu GC-2010Pro (Japan), a quartz capillary column (SHRT-2560, 100 m.times.0:25 mm.times.0:20 μm) and equipped with a flame ionization detector. The column temperature was programmed to rise from 150 ℃ to 200 ℃ at 15 ℃ per minute and then from 200 ℃ to 250 ℃ at 2 ℃ per minute. The sample inlet and detector temperatures were 250 ℃. The fatty acid is expressed in terms of the ratio of certain fatty acids to 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 method. Data are expressed as mean ± standard error (n=4). The difference is indicated as significant with P < 0.5.
5. Experimental results
The experiment mainly evaluates the application effect of the technology according to the survival rate, the weight gain rate and the muscle fatty acid composition.
In the experimental process, the final survival rates are not significantly different and are all above 90% (figure 1). The weight gain rate was not significantly different from that of the fish oil control group (group a) using the fatty acid balance group (group B) of the present invention (P > 0.05, fig. 2). The high 18C-PUFA group (group E) had a significantly lower weight gain than the fatty acid balance group (group B) and the fish oil control group (group a) (P < 0.05) without significant difference from the high MUFA group (group D). The high SFA group (group C) has the lowest weight gain rate, which is significantly lower than the other groups (P < 0.05).
In terms of muscle fatty acid composition (table 3), groups a and B obtained similar fatty acid compositions. Especially in terms of long chain polyunsaturated fatty acids, group B only achieved slightly lower ARA, EPA and DHA content than the fish oil control group (group a). The fatty acid composition of group B is shown to be very balanced, and long-chain polyunsaturated fatty acids such as ARA, EPA and DHA can be furthest reserved for muscle accumulation. While the amounts of ARA, EPA and DHA were reduced to varying degrees in the three groups C, D and E. B. C, D and group E muscle DHA content was 94.3%, 59.7%, 52.3% and 49.1% respectively for group a (fig. 3); B. c, D and group E muscle EPA content was 90.7%, 22.7%, 35.8% and 51.4% of group a, respectively (fig. 3); B. c, D and E group muscle ARA levels were 97.2%, 93.2%, 86.4% and 90.4%, respectively, for group A (FIG. 3).
Table 3 turbot muscle fatty acid composition (% total fatty acid, mean ± standard error, n=4)
There was a significant difference (P < 0.05) between data columns in the same row that did not contain the same letter. SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
Example 2 evaluation test of the Effect of the method on Fugu rubripes cultivation
1. Experimental design and Experimental feed formulation (basic feed formulation is a commercial feed formulation which simulates common use, and does not limit the scope of protection of the invention, and the effect of the invention can be achieved by implementing the nutritional method of the invention under the condition that the cultured fish can grow)
A feed fatty acid balancing technology and its application in sea fish, its method is as follows:
for fish with muscle polar lipid content higher than 70% of total lipid content (Takifugu rubripes Takifugu rubripes in this example), the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5:3:1.2:1. Wherein SFA (mainly comprising 14:0, 16:0, 18:0 and 20:0), MUFA (mainly comprising 16:1n-7, 18:1n-9, 20:1n-9 and 22:1 n-11), 18C-PUFA (mainly comprising 18:2n-6 and 18:3 n-3) and LC-PUFA (mainly comprising 20:2n-6, 20:3n-3, ARA, EPA, DPA and DHA) account for 25%, 30%, 12% and 10% of total fatty acids, respectively.
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 collocation schemes to obtain different fatty acid compositions (with SFA/MUFA/18C-PUFA/LC-PUFA ratio as core index) and the fish oil group as control group (Table 4). The feeds for each group were designated A (fish oil control group), B (fatty acid balance group), C (high SFA group), D (high MUFA group) and E (high 18C-PUFA group), respectively. The fatty acid test results for each feed group are shown in table 5.
Table 4 feed formulation and coarse ingredients (% dry matter) of experimental feed
Table 5 Experimental feed fatty acid composition (% Total fatty acids)
SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Experimental fish and aquaculture management
The experiment adopts the young fish of the fugu rubripes with the initial weight of 12g, and before the formal experiment, the experimental fish is temporarily cultured in a cement pond with the square meter of 25 ℃ for 10 days so as to adapt to the experimental environment conditions. Before the beginning of the formal experiment, the experimental fish were randomly divided into 20 polyethylene barrels (height: 100cm; diameter: 230 cm), each group was repeated 4 times, 35 fish per barrel. Indoor running water culture is adopted in the culture process, and underground deep well water is adopted as the seawater for culture. Fed by manual satiety for 2 times each day. The culture experiments were carried out at the Shaoyang yellow sea aquatic products Limited company (flounder breeding base) of yellow sea aquatic products institute of China aquatic science, and the culture period was 8 weeks. The cultivation experiments were performed at natural photoperiod and ambient temperature (N36°41', E121°07') in the sea-yang city of shandong province, china. In the feeding and breeding experiment process, the water temperature range is 14-18 ℃; salinity, 29-31; the pH is 7.2-8.4; dissolved oxygen, 6-8 mg L -1 . Residual feed and faeces were removed by siphoning half an hour after the end of daily feeding.
3. Terminal body weight measurement, sample collection and fatty acid analysis
After the cultivation experiment is finished, the total weight of each barrel of fish is weighed, and the total weight is counted to calculate the survival rate. Meanwhile, 5 fish are dissected, the weights of livers and viscera are weighed, and the body length is measured and used for calculating the liver-body ratio, the viscera-body ratio and the fullness. The calculation method of the liver body ratio, the viscera body ratio and the fullness comprises the following steps: liver ratio (%) = liver weight (fresh weight)/body weight×100; visceral volume ratio (%) = visceral weight (fresh weight)/body weight x 100; fullness = body weight (g)/body length (cm) 3 . A muscle (dorsum muscle) sample of 6 fish was taken and the fatty acid content was determined as follows: esterifying the freeze-dried sample with KOH-methanol and HCl-methanol respectivelyWater bath at 72 ℃), and then methyl esterified fatty acids were extracted with n-hexane, followed by measurement on the machine. Gas chromatography was carried out using Shimadzu GC-2010Pro (Japan), a quartz capillary column (SHRT-2560, 100 m.times.0:25 mm.times.0:20 μm) and equipped with a flame ionization detector. The column temperature was programmed to rise from 150 ℃ to 200 ℃ at 15 ℃ per minute and then from 200 ℃ to 250 ℃ at 2 ℃ per minute. The sample inlet and detector temperatures were 250 ℃. The fatty acid is expressed in terms of the ratio of certain fatty acids to 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 method. Data are expressed as mean ± standard error (n=4). The difference is indicated as significant with P < 0.5.
5. Experimental results
The experiment mainly evaluates the application effect of the technology according to the survival rate, the weight gain rate and the muscle fatty acid composition.
In the experimental process, the final survival rates were not significantly different, and were all above 84% (fig. 4). The weight gain rate was not significantly different from that of the fish oil control group (group A) using the fatty acid balance group (group B) of the present invention (P > 0.05, FIG. 5). The high MUFA group (group D) and the high 18C-PUFA group (group E) had significantly lower weight gain than 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 the other groups.
Very similar to the results on turbots, groups a and B obtained similar fatty acid compositions in terms of muscle fatty acid composition (table 6). Especially in terms of long chain polyunsaturated fatty acids, group B only achieved slightly lower ARA, EPA and DHA content than the fish oil control group (group a), which indicated that the fatty acid composition of group B was very balanced, and long chain polyunsaturated fatty acids such as ARA, EPA and DHA could be retained to the maximum extent for muscle accumulation. In particular the DHA and EPA content, there was no significant difference between groups A and B. While the amounts of ARA, EPA and DHA were reduced to varying degrees in the three groups C, D and E. B. C, D and group E muscle DHA content was 90.6%, 69.3%, 60.9% and 64.6% respectively for group a (fig. 6); B. c, D and group E muscle EPA content were 86.7%, 55.5%, 70.3% and 43.3% of group a, respectively (fig. 6); B. c, D and E group muscle ARA levels were 75.8%, 66.7% and 69.7%, respectively, for group A (FIG. 6).
Table 6 experiment of fatty acid composition of fugu rubripes muscle (% total fatty acid, average ± standard error, n=4)
There was a significant difference (P < 0.05) between data columns in the same row that did not contain the same letter. SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
Example 3 evaluation of Effect of turbot Pilot Scale experiment Using the method
1. Experimental design and Experimental feed formulation (basic feed formulation is a commercial feed formulation which simulates common use, and does not limit the scope of protection of the invention, and the effect of the invention can be achieved by implementing the nutritional method of the invention under the condition that the cultured fish can grow)
A feed fatty acid balancing technology and its application in sea fish, its method is as follows:
for fish with muscle polar lipid content lower than 70% of total lipid content (the experimental animal in this experiment is turbot 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. Wherein SFA (mainly comprising 14:0, 16:0, 18:0 and 20:0), MUFA (mainly comprising 16:1n-7, 18:1n-9, 20:1n-9 and 22:1 n-11), 18C-PUFA (mainly comprising 18:2n-6 and 18:3 n-3) and LC-PUFA (mainly comprising 20:2n-6, 20:3n-3, ARA, EPA, DPA and DHA) account for 25%, 15% and 10% of total fatty acids, respectively.
The experimental feeds used in this example were only group a and group B of the examples due to pilot scale experimental site limitations. The experimental feed formulation and method of manufacture 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. The crude fat content was about 10-11%, and the experimental feeds of group 2 used different fat source collocation schemes to obtain different fatty acid compositions, with the fish oil group as the control group (group a) and the fatty acid balance group using the method as the experimental group (group B) (table 7). The fatty acid test results of the two groups of experimental feeds are shown in table 8.
Table 7 feed formulation and coarse ingredients (% dry matter) of experimental feed
Table 8 Experimental feed fatty acid composition (% Total fatty acids)
SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Experimental fish and aquaculture management
The experiment adopts experimental fish with initial weight of 152g, 2 groups of experimental feeds are respectively fed into 2 indoor mariculture cement ponds (3 m multiplied by 1.5 m), 300 experimental fish are placed in each cement pond, the culture experiment is carried out in yellow sea aquatic products limited company in Haiyang city of smoke table, and the culture water is underground deep well seawater, and running water culture is adopted. Fed twice daily with a total cultivation period of 10 weeks (70 days).
3. Terminal body weight measurement, sample collection and fatty acid analysis
After the cultivation experiment is finished, 50 fish are randomly sampled in each cement pond, weighed and the weight gain rate is calculated. Meanwhile, muscle (dorsal muscle) samples of 10 fish were randomly taken per cage, and the fatty acid content was determined as follows: the lyophilized samples were esterified with KOH-methanol and HCl-methanol, respectively (72 ℃ C. Water bath), and the methyl-esterified fatty acids were extracted with n-hexane and then measured on an engine. Gas chromatography was carried out using Shimadzu GC-2010Pro (Japan), a quartz capillary column (SHRT-2560, 100 m.times.0:25 mm.times.0:20 μm) and equipped with a flame ionization detector. The column temperature was programmed to rise from 150 ℃ to 200 ℃ at 15 ℃ per minute and then from 200 ℃ to 250 ℃ at 2 ℃ per minute. The sample inlet and detector temperatures were 250 ℃. The fatty acid is expressed in terms of the ratio of certain fatty acids to total fatty acids.
4. Experimental statistical method
Since no duplicate cement reservoirs were designed, only the mean and standard deviation were calculated and no statistical analysis was performed.
5. Experimental results
In terms of survival rates, the survival rates for groups a and 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 rate of gain (fig. 8). In terms of muscle long chain polyunsaturated fatty acid content (fig. 9), the DHA, EPA and ARA content of group B were 96%, 74.6% and 169%, respectively, of group a. Group B had even more ARA content than group a.
Example 4 evaluation of the Effect of the application of the method in pilot-scale experiments on Fugu rubripes
1. Experimental design and Experimental feed formulation (basic feed formulation is a commercial feed formulation which simulates common use, and does not limit the scope of protection of the invention, and the effect of the invention can be achieved by implementing the nutritional method of the invention under the condition that the cultured fish can grow)
A feed fatty acid balancing technology and its application in sea fish, its method is as follows:
for fish with muscle polar lipid content higher than 70% of total lipid content (Takifugu rubripes Takifugu rubripes in this example), the ratio of SFA/MUFA/18C-PUFA/LC-PUFA in the feed is controlled as follows: 2.5:3:1.2:1. Wherein SFA (mainly comprising 14:0, 16:0, 18:0 and 20:0), MUFA (mainly comprising 16:1n-7, 18:1n-9, 20:1n-9 and 22:1 n-11), 18C-PUFA (mainly comprising 18:2n-6 and 18:3 n-3) and LC-PUFA (mainly comprising 20:2n-6, 20:3n-3, ARA, EPA, DPA and DHA) account for 25%, 30%, 12% and 10% of total fatty acids, respectively.
The same feed formulation as in example 2 was used in this experiment, with only A, B treatment groups due to pilot scale limitations. Namely, 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 group 2 experimental feeds used different fat source collocation schemes to obtain different fatty acid compositions (with SFA/MUFA/18C-PUFA/LC-PUFA ratio as core index), the group of fish oils as control group (group A), and the group of fatty acid balanced feeds using the method of the present invention as treatment group (group B) (Table 9). The feed fatty acid composition is shown in table 10.
Table 9 feed formulation and crude ingredients (% dry matter) of experimental feeds
Table 10 Experimental feed fatty acid composition (% Total fatty acids)
SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
2. Experimental fish and aquaculture management
The experiment adopts experimental fish with initial weight of 109g, 2 groups of experimental feeds are respectively fed into 2 mariculture net cages (3 m multiplied by 2 m), 400 experimental fish are placed in each net cage pool, and the culture experiment is carried out in Tangshan city in Hebei. Fed twice daily with a total cultivation period of 12 weeks (84 days).
3. Terminal body weight measurement, sample collection and fatty acid analysis
After the cultivation experiment is finished, 50 fish are randomly sampled in each cement pond, weighed and the weight gain rate is calculated. Meanwhile, muscle (dorsal muscle) samples of 10 fish were randomly taken per cage, and the fatty acid content was determined as follows: the lyophilized samples were esterified with KOH-methanol and HCl-methanol, respectively (72 ℃ C. Water bath), and the methyl-esterified fatty acids were extracted with n-hexane and then measured on an engine. Gas chromatography was carried out using Shimadzu GC-2010Pro (Japan), a quartz capillary column (SHRT-2560, 100 m.times.0:25 mm.times.0:20 μm) and equipped with a flame ionization detector. The column temperature was programmed to rise from 150 ℃ to 200 ℃ at 15 ℃ per minute and then from 200 ℃ to 250 ℃ at 2 ℃ per minute. The sample inlet and detector temperatures were 250 ℃. The fatty acid is expressed in terms of the ratio of certain fatty acids to total fatty acids.
4. Experimental statistical method
Since no duplicate cage was designed, only the mean and standard deviation were calculated and no statistical analysis was performed.
5. Experimental results
In terms of survival rate, the survival rates of the group A and the group B are 81.3% and 85.5% respectively, and the survival rate of the group B is even slightly higher than that of the group A (figure 10). Group B (368%) was also slightly higher than group a (324%) in terms of rate of weight gain (fig. 11). In terms of muscle long chain polyunsaturated fatty acid content (fig. 12), the DHA, EPA and ARA content of group B were 90.2%, 115.6% and 106.3% respectively of group a. Both EPA and ARA contents of group B exceeded those of group A.
Claims (3)
1. A method for balancing fatty acid in fish feed, which is characterized by comprising the following steps:
(1) For fish with muscle polar lipid content 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 control feed is as follows: 2.5:2.5:1.5:1; wherein the mass percentages of saturated fatty acid, monounsaturated fatty acid, 18-carbon polyunsaturated fatty acid and long-chain polyunsaturated fatty acid in the total fatty acid are 25.46%, 25.3%, 15.46% and 11.87%, respectively;
(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 control feed is as follows: 2.5:3:1.2:1; wherein the mass percentages of saturated fatty acid, monounsaturated fatty acid, 18-carbon polyunsaturated fatty acid and 18-carbon polyunsaturated fatty acid in the total fatty acid are 25.3%, 30.24%, 12.36% and 10.83%, respectively;
the content of the saturated fatty acid is detected to be 14:0, 16:0, 18:0 and 20:0;
the content of the monounsaturated fatty acid is 16:1n-7, 18:1n-9, 20:1n-9 and 22:1n-11;
the content of the 18 carbon polyunsaturated fatty acid is the detected content of 18:2n-6 and 18:3n-3;
the content of the long-chain polyunsaturated fatty acid is 20:2n-6, 20:3n-3, ARA, EPA, DPA and DHA.
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 for the cultivation of fish.
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