CN107980680B - High-value utilization method of haematococcus pluvialis residues in economic crab fattening feed - Google Patents
High-value utilization method of haematococcus pluvialis residues in economic crab fattening feed Download PDFInfo
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- CN107980680B CN107980680B CN201711408090.8A CN201711408090A CN107980680B CN 107980680 B CN107980680 B CN 107980680B CN 201711408090 A CN201711408090 A CN 201711408090A CN 107980680 B CN107980680 B CN 107980680B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/59—Culture of aquatic animals of shellfish of crustaceans, e.g. lobsters or shrimps
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- 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
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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
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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
Abstract
The invention discloses a high-value utilization method of haematococcus pluvialis residues in economic crab fattening feed, which comprises the following steps of: (1) measuring the content of astaxanthin in the haematococcus pluvialis residues; (2) determining the addition amount of the haematococcus pluvialis residues in different economic crab fattening feed formulas; (3) processing the economic crab fattening feed by using a suitable feed processing technology; and (4) scientifically feeding the economic crabs with fattening feed for fattening. The method comprises the steps of calculating the addition amount of haematococcus pluvialis residues in a feed formula according to the astaxanthin demand of the crabs during fattening and the astaxanthin content in the haematococcus pluvialis residues; the astaxanthin oxidation loss in the feed is prevented by adopting a grease vacuum spraying process; scientifically fattening according to the development condition of the ovary of the crab. The fattening feed prepared and used by the method can reduce the feed formula cost by more than 1500 yuan/ton, improve the fattening effect of the crabs, and lead the red cream rate after fattening to be more than 85 percent.
Description
Technical Field
The invention relates to a high-valued application method of haematococcus pluvialis residues in crab fattening feed, and belongs to the technical field of aquatic feed and cultivation.
Background
Haematococcus pluvialis (Haematococcus pluvialis), an important freshwater economic green alga, has a dry powder content of usually 1.5-6% of natural astaxanthin, is a main source of natural astaxanthin, and has a high economic value (Sandeshiet al, 2008; Orosa et al, 2005). The natural astaxanthin in haematococcus pluvialis powder is all-trans-levo-astaxanthin, exists in an esterified form, has stable properties and higher bioavailability (Holtin et al, 2009); in addition, natural astaxanthin has important functions of inhibiting tumor cell proliferation (Cerariu et al, 2015), improving oxidation resistance and immunity (Fiedor and Durdura, 2014; Hughes,2001), resisting aging (Kidd,2011), reducing the risk of cancer and cardiovascular diseases (Shareck et al, 2017; Riccioni,2009), and the like. Thus, Haematococcus pluvialis-derived natural astaxanthin is mainly used in health products, foods, cosmetics and natural drugs (Tominaga et al, 2012; Palozza et al, 2009). The haematococcus pluvialis is usually generated into a large amount of haematococcus pluvialis residues after astaxanthin is extracted, and the extraction process is usually supercritical extraction by adopting food-grade ethanol or carbon dioxide solvent, so that the ethanol or carbon dioxide is only required to be volatilized and removed from the haematococcus pluvialis residues after the astaxanthin is extracted, and the haematococcus pluvialis residues are nontoxic and harmless and can be largely utilized on animal feed; research shows that the haematococcus pluvialis residue after astaxanthin extraction contains conventional nutritional ingredients such as protein, fat and carbohydrate, and also contains a certain amount of active substances such as astaxanthin and immune polysaccharide, so that the haematococcus pluvialis residue can improve animal color and luster, and improve immune disease resistance and nutritional quality, and can be used as a high-grade feed raw material to be used in aquatic feeds at a high value (Ju et al, 2012; rolling stocks and the like, 2017). At present, haematococcus pluvialis residues are mainly used as feed raw materials to be applied to livestock and poultry breeding, the price is only 60-100 yuan/kg, and the application to aquatic feeds is less, and a utilization approach and a method in crab fattening feeds are not seen yet.
The crab meat is generally required to be fed with high-quality feed (bait) to ensure the normal development of the crab, for example, the crab meat requires high-quality feed (bait) to ensure the normal development of the crab meat in the gonad development stage, for example, the feed contains high contents of protein, fat, vitamins, astaxanthin and the like, the process of promoting the gonad development is called fattening, the bait fed in the gonad development stage is called bait, the crab meat is called Zhao bait, the content of astaxanthin in the crab meat feed is very important for the color, the oxidation resistance, the immunity and the nutritional composition, so that the crab meat feed generally needs to be added with a certain content of astaxanthin (L ong et al, 2017; epididymis, 2017) to feed the crab meat feed in a certain amount of astaxanthin-containing natural astaxanthin in the gonad development stage to ensure the normal development of the crab meat, the natural astaxanthin, the immune disease resistance, the natural astaxanthin and the nutritional composition, the bait are added in the crab meat feed for fattening pond, the crab meat feed, the natural astaxanthin, the bait is usually enough for the natural astaxanthin, the crab meat, the bait is added with high-resistant natural astaxanthin, the bait is added in the bait, the bait is added in the bait, the natural astaxanthin, the bait, the natural astaxanthin, the bait, the natural astaxanthin, the bait, the natural astaxanthin, the bait, the natural astaxanthin, the bait.
Based on the research and development of crab nutritional feeds and fattening technologies, the inventor finds that haematococcus pluvialis residues can be applied to crab fattening feeds and can replace expensive haematococcus pluvialis powder or artificially synthesized astaxanthin, so that the cost of the crab fattening feeds can be greatly reduced, the immunity and the nutritional quality of crabs can be improved, and the crab fattening feeds have important application value. Therefore, the application of haematococcus pluvialis residues in the female sports fertilizer feed for eriocheir sinensis is taken as an example in the invention to illustrate the feasibility of the invention.
Disclosure of Invention
On the basis of a great deal of research, according to the principle that the algae residue obtained after astaxanthin is extracted from haematococcus pluvialis powder still contains a certain amount of active substances such as astaxanthin, immune polysaccharide and the like, and according to the requirements of astaxanthin and other nutrients of economic crabs during gonad development, a certain amount of haematococcus pluvialis algae residue is added into the fattening feed, so that the color and luster of river crabs can be improved, the oxidation resistance and the immunity can be improved, the content of astaxanthin in edible tissues can be improved, the high-valued utilization of the haematococcus pluvialis algae residue can be realized, and the formula cost of the crab fattening feed can be greatly reduced. The invention aims to provide a high-value utilization method of haematococcus pluvialis residues in crab fattening feed. In order to realize the purpose of the invention, the technical scheme of the invention is as follows:
a high-value utilization method of haematococcus pluvialis residues in economic crab fattening feed comprises the following steps:
(1) measuring the content of astaxanthin in the haematococcus pluvialis residues;
(2) determining the addition amount of the haematococcus pluvialis residues in different economic crab fattening feed formulas;
(3) processing the economic crab fattening feed by using a suitable feed processing technology; and
(4) and scientifically feeding the economic crabs with fattening feed for fattening.
The crabs economical in the invention comprise Chinese mitten crabs, portunus trituberculatus, scylla paramamosain and American blue crabs. A
In a preferred embodiment of the present invention, the haematococcus pluvialis algal residue is haematococcus pluvialis algal residue generated after the astaxanthin is extracted by the haematococcus pluvialis with ethanol or a carbon dioxide solvent in a supercritical manner.
In a preferred embodiment of the present invention, the step (1) of determining the astaxanthin content in the algal residue of Haematococcus pluvialis comprises the steps of: extracting the Haematococcus pluvialis residue with acetone as solvent, and determining astaxanthin content by liquid chromatography.
In a more preferred embodiment of the invention, the chromatographic column used in the liquid chromatography is L una3u silica, and the mobile phase is n-hexane and acetone in a volume ratio of 83: 17.
In a preferred embodiment of the invention, the addition amount of the haematococcus pluvialis residues in different economic crab fattening feed formulas determined in the step (2) is calculated according to the demand amount of astaxanthin during gonad development (fattening period) of female and male economic crabs and the content of astaxanthin in the haematococcus pluvialis residues, and the addition amount of the haematococcus pluvialis residues and other raw material formulas in corresponding economic crab fattening feed formulas.
In a preferred embodiment of the present invention, the suitable feed processing process in step (3) comprises a sinking expanded feed processing process and oil vacuum spraying to the surface of the economic crab fattening feed to ensure the water stability of the economic crab fattening feed and prevent the oxidation loss of astaxanthin therein.
In a more preferred embodiment of the present invention, the suitable processing in step (3) comprises the steps of:
(1) various economic crab fattening feed raw materials are subjected to superfine grinding, mixing and tempering to prepare sinking expanded feed, and the sinking expanded feed is prepared into particles;
(2) spraying grease on economic crab fattening feed particles;
(3) double-layer packaging, wherein the inner layer is a vacuumized plastic bag, and the outer layer is a dark plastic woven bag;
(4) the feed is stored in dark place and stored in a cool and dry place or a refrigerator.
In a further preferred embodiment of the invention, the fat is a mixture of fish oil and plants in an amount of 4-8% by weight of the total feed.
In a preferred embodiment of the invention, scientific feeding of the economic crabs for fattening in the step (4) is carried out, and more than 80% of individuals should complete the genital molting, so that feeding of the fattening feed for the economic crabs can be started.
In a more preferred embodiment of the invention, in the step (4), the economic crabs are fattened by scientifically feeding fattening feed, the female and male economic crabs are recommended to be fattened by being separately fed, the ovary index of the female crabs is 0.5-3% before fattened, the ovary index is more than 7% after fattened, and fattened is stopped; the gonadal index before fattening the male crab is about 0.5 percent.
In a more preferred embodiment of the invention, when the male and female economic crabs are mixed and cultured, female sports fat feed is fed to fatten the male and female economic crabs mixed and cultured.
On the one hand, on the basis of measuring astaxanthin and other nutritional ingredients in haematococcus pluvialis residues and combining with nutritional requirements in different crab fattening periods, the crab fattening feed with balanced nutrition and lower cost is prepared, and the cost is reduced by more than 1500 yuan/ton compared with the cost of a feed formula adopting natural astaxanthin or artificially synthesized astaxanthin; the second aspect of the invention provides a method for using crab fattening feed (containing haematococcus pluvialis residues), which can improve the color and luster and the astaxanthin content in tissues, enhance the immunity and disease resistance and improve the nutritional quality, thereby improving the economic value of the fattening crabs and realizing the high-value utilization of the haematococcus pluvialis residues.
The technical scheme of the invention is suitable for haematococcus pluvialis culture and processing enterprises, feed processing enterprises, crab culture enterprises or farmers, is beneficial to improving the color of cultured river crabs, improving the immunity, disease resistance and stress resistance, and simultaneously reducing the bait cost and culture risk.
Astaxanthin and other main nutrient content determination of haematococcus pluvialis residue。
The method comprises the steps of extracting total carotenoids in algal residues by using acetone as a solvent, measuring the astaxanthin content by using a liquid chromatography, measuring the astaxanthin content by using L una3u Silica (150mm × 4.60.60 mm, phenomenex, USA) as a chromatographic column, measuring the feed water content (105 ℃ drying method), the crude protein (Kjeldahl method) and the crude ash content (550 ℃ burning method) by using normal hexane and acetone (TUme et al,2009) according to the standard method of AOAC, extracting the total lipids by using chloroform-methanol (V/V ═ 2:1) and measuring the content (Folch et al, 1957), carrying out methyl esterification on the total lipids by using a 14% boron trifluoride-methanol solution, analyzing the fatty acid composition by using Agilent 7890B-5977A gas chromatography-mass spectrometer, measuring the chromatographic column by using Agilent SP-2560 capillary column (100m × 0.25.25 mm; 0.2 μm) as a specific method, referring to dry cysteine, measuring the methionine content by using hydrochloric acid hydrolysis method (1982007), measuring the amino acid content by using hydrochloric acid hydrolysis method, and the general method (Spinden hydrolysis method).
Determination of nutrition parameters of feed in crab fattening stage (gonadal development stage)
According to the research experience of the inventor and the consulted data, the nutritional parameters of the feed for the crab male and female individuals in the fattening stage (gonad development stage) are determined. Taking river crabs as an example, the main nutritional requirement parameters during the development of female ovaries are as follows: 34-38% of crude protein, 10-14% of crude fat, 3-4% of phospholipid, 0.5-0.8% of total height unsaturated fatty acid and about 60mg/kg of trans-L-astaxanthin; the main nutritional requirement parameters during development of androgenic glands are: 37-40% of crude protein, 6-8% of crude fat, 2-3% of phospholipid, 0.3-0.5% of total height unsaturated fatty acid and 30mg/kg of trans-L-astaxanthin.
Fattening feed formula and determination of addition amount of haematococcus pluvialis residues
According to the nutritional requirements, the raw material characteristics and the formula experience of the crab fattening stage, the main raw material composition and the addition amount of haematococcus pluvialis residues in the feed formula are determined. For example, the requirement of the female river crab on trans-astaxanthin is 60mg/kg of feed, for example, the content of the trans-astaxanthin in the basic feed is about 10mg/kg of feed, and the content of the trans-astaxanthin in the haematococcus pluvialis algae residue is 0.6 percent, so that the haematococcus pluvialis algae residue which is 0.83 percent needs to be added into the female river crab sports fat feed can be calculated according to the requirement. The other main components in the female sports fertilizer feed are as follows: 10-15% of imported steam fish meal, 5-8% of meat meal, 18-24% of fermented soybean meal, 10% of peanut meal, 10-12% of rapeseed meal, 8-12% of corn, 14% of wheat middling, 4% of antarctic krill meal, 3% of soybean lecithin oil, 2.0% of refined fish oil, 2% of lard, 1% of soybean oil, 1% of rapeseed oil, 1% of monocalcium phosphate, 0.5% of choline chloride, 0.2% of compound vitamin, 0.3% of cholesterol, 0.5% of compound mineral, 0.2% of betaine and 50mg/kg of vitamin E.
Fattening feedIs processed by
Various feed raw materials are subjected to superfine grinding (all raw materials pass through a 100-mesh screen), mixing and tempering to prepare sinking expanded feed, and in order to prevent the oxidation loss of astaxanthin added in the feed, 4-8% of oil is sprayed on the surface of the feed by adopting a vacuum spraying process after the expanded feed is granulated, so that an oxygen isolation layer can be formed to prevent the oxidation of astaxanthin. The grain size and the length of the fattening feed are determined according to the breeding objects; the expanded feed has good stability in water, and the dissolution rate of the expanded feed soaked in water for 4 hours (under the condition of 20 ℃) is less than 10%.
Packaging and preservation of fattening feed
Because the crab fattening feed contains higher astaxanthin and highly unsaturated fatty acid, the crab fattening feed usually needs to be vacuumized and packaged, the outer layer is a dark color plastic woven bag to prevent the oxidative decomposition of the fatty acid and the astaxanthin caused by the feed after being exposed to light, the feed packaging bag is usually two layers, the inner layer is a vacuumized plastic bag, and the outer layer is a dark color woven bag; the feed is stored in a cool and dry place or a refrigerator with the temperature below-10 ℃.
Using method of fattening feed
After more than 80% of individuals complete reproductive molting, feeding fattening feed, separately feeding female and male crabs for fattening, wherein the ovary index of the female crabs before fattening is 0.5-3% (the fattening effect is poor when the ovary index is too large), and the ovary index is more than 7% after fattening, so that the fattening can be stopped; if the fattening crab is male crab, the gonad index before fattening is about 0.5%. During mixed culture of the female and the male, the nutritional requirements of the female to the astaxanthin are higher than those of the male, and the nutritional requirements of the male can be met by the female sports fertilizer feed, so that the female sports fertilizer feed is generally fed during mixed culture. The special feed for the fattening stage is fed twice a day, wherein the feeding amount in the morning accounts for 30-40% of the total weight of the river crabs (the water temperature in a pond is 20-30 ℃), the feeding time is about 7:00 in the morning, and the feeding in a deep water area is mainly used; the feeding amount in the afternoon accounts for 60-70% of the total day, the feeding time is about 18:00 in the afternoon, and the feeding is mainly carried out in shallow water. The daily average water temperature of the pond is lower than 20 ℃, the daily feeding amount is usually about 0.5-2% of the total weight of the river crabs, the pond is fed for 0.5-1 time every day, and the feeding time is about 17:00 in the afternoon.
The principle of the invention is as follows:
haematococcus pluvialis residues are byproducts of natural astaxanthin extraction, and are mainly used for livestock feed or as fertilizer at present. The research result of the inventor shows that the haematococcus pluvialis residue generally contains 25-30% of crude protein, 5-12% of crude fat, 25-30% of carbohydrate and 0.2-1% of astaxanthin; since crabs require a higher demand for astaxanthin during gonadal development, but the use of haematococcus pluvialis powder or artificially synthesized astaxanthin as a source of astaxanthin in the feed is very costly. The research of the inventor shows that the haematococcus pluvialis residues are used as the main source of astaxanthin in the crab fattening feed, so that the feed cost can be obviously reduced, the high-value utilization of the haematococcus pluvialis residues can be realized, and the immune disease resistance and the nutritional quality of the crabs can be improved. Repeated practice in recent years shows that the cost of the crab fattening feed prepared by applying the method of the invention is reduced by about 1500 yuan/ton compared with the crab fattening feed prepared by adopting haematococcus pluvialis powder or artificially synthesized astaxanthin as an astaxanthin source, the survival rate of the crabs is more than 80 percent after fattening for 40-60 days, and the red paste rate is more than 85 percent. The invention has convenient application, good practical use effect and stronger practical value. Due to the adoption of the technical scheme, the invention has the following beneficial effects:
1. according to the nutritional composition of the haematococcus pluvialis residues and the nutritional requirements during crab fattening, the haematococcus pluvialis residues are applied to crab fattening feeds to realize high-value utilization of the haematococcus pluvialis residues;
2. the haematococcus pluvialis residues are used for replacing haematococcus pluvialis powder or artificially synthesized astaxanthin which are commonly used in the conventional crab fattening feed, so that the cost of the feed can be greatly reduced under the condition of ensuring the fattening effect, and the possible negative influence of formal astaxanthin existing in the artificially synthesized astaxanthin on animal bodies or human bodies can be prevented.
3. According to different nutritional requirements, reproductive molting time, fattening period and the like of crab male and female individuals during gonad development period (fattening period), the fattening feed special for male and female individuals is prepared respectively, so that the fattening effect can be improved, and accurate fattening is realized.
4. Feeding fattening feed rich in trans-astaxanthin for 40-60 days at the crab gonad development stage, and increasing the content of astaxanthin and the like in crab shell, ovary and liver and pancreas, thereby improving the color of the crab shell, ovary and liver and pancreas and increasing the commercial value and the edible value of the crabs;
5. by adding sufficient trans-astaxanthin into the feed, the color and luster can be regulated and controlled, the nutritional value can be improved, and the immunity and stress resistance of the river crabs can be improved, so that the survival rate of the river crabs in the fattening period is improved.
Drawings
Figure 1 is the effect of three feeds on the ovarian index of river crabs.
FIG. 2 shows the influence of three kinds of feed on the color of dried river crab ovary. The feed 1# group is river crabs fed with the feed added with haematococcus pluvialis residues, the ovary color of female river crabs in the feed 1# group after being dried is ruddy, and is close to that of the feed 2# group (river crabs fed with the feed added with haematococcus pluvialis powder); the feed No. 2 is river crabs fed with the feed added with the wall-broken haematococcus pluvialis powder, and the color of ovaries of the female river crabs in the group after drying is good; the feed No. 3 group is river crabs (as a control group) which are fed with the feed without adding haematococcus pluvialis algae residues or algae powder, and the ovary color of the female river crabs in the group after being dried is light yellow, and is obviously different from that of the feed No. 1 group and the feed No. 2 group.
FIG. 3 shows the influence of three kinds of feed on the color of dried crab shell. The color of the dried crab shells of the feed 1# group and the feed 2# group is dark orange red, and the color of the dried crab shells of the feed 3# group (the fattening feed without adding haematococcus pluvialis residue or algae powder) is light orange red, which is obviously different from that of the feed 1# group and the feed 2# group.
Detailed description of the preferred embodiment
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further illustrated by the application example of the haematococcus pluvialis residues on the female sports fat feed of the river crabs.
1.1 astaxanthin from Haematococcus pluvialis residues and use thereofDetermination of main nutrient components of the medicine
The method comprises the steps of extracting total carotenoids in algal residues by using acetone as a solvent, measuring the astaxanthin content by using a liquid chromatography, measuring the astaxanthin content by using L una3u Silica (150mm × 4.60.60 mm, phenomenex, USA) as a chromatographic column, measuring the feed water content (105 ℃ drying method), the crude protein (Kjeldahl method) and the crude ash content (550 ℃ burning method) by using normal hexane and acetone (TUme et al,2009) according to the standard method of AOAC, extracting the total lipids by using chloroform-methanol (V/V ═ 2:1) and measuring the content (Folch et al, 1957), carrying out methyl esterification on the total lipids by using a 14% boron trifluoride-methanol solution, analyzing the fatty acid composition by using Agilent 7890B-5977A gas chromatography-mass spectrometer, measuring the chromatographic column by using Agilent SP-2560 capillary column (100m × 0.25.25 mm; 0.2 μm) as a specific method, referring to dry cysteine, measuring the methionine content by using hydrochloric acid hydrolysis method (1982007), measuring the amino acid content by using hydrochloric acid hydrolysis method, and the general method (Spinden hydrolysis method).
TABLE 1 astaxanthin and other major nutritional components of Haematococcus pluvialis algal lees
∑ EAA total essential amino acid content, ∑ NEAA total nonessential amino acid content, ∑ TAA total amino acid content, ∑ SFA total saturated fatty acid content, ∑ MUFA total monounsaturated fatty acid content, ∑ PUFA total polyunsaturated fatty acid content, ∑ HUFA total unsaturated fatty acid content.
1.2 determination of nutritional parameters of female stage of physical fattening (gonadal development stage) feed for river crab
The inventor researches the experience and consults the data to determine that the main nutritional parameters of the female river crab in the fattening stage (gonad development stage) to the feed are as follows: 34-38% of crude protein, 10-14% of crude fat, 3-4% of phospholipid, 0.5-0.8% of total height unsaturated fatty acid and about 60mg/kg of trans-L-astaxanthin.
1.3 fattening feed formulation and determination of addition amount of Haematococcus pluvialis residue
According to the nutritional requirements, the raw material characteristics and the formula experience of female sports fertilizers of the river crabs, the main raw material composition and the addition amount of haematococcus pluvialis residues in the feed formula are determined. The river crab fattening feed takes bean pulp, rapeseed meal and fish meal as main protein sources, takes fish oil, lard, soybean oil and rapeseed oil as fat sources to prepare experimental feed, and in order to scientifically evaluate the influence of haematococcus pluvialis residues on female fattening and nutritional quality of river crabs, two control groups (a haematococcus pluvialis powder group and a control group without haematococcus pluvialis powder or algae residues) are arranged in the experiment, so that three groups of feeds are total: feed 1# (1% of haematococcus pluvialis residue is added, and the actually measured astaxanthin content in the algae residue is about 0.66%); feed # 2 (0.2% of wall-broken haematococcus pluvialis powder is added, and the actually measured astaxanthin content in the haematococcus pluvialis powder is about 3.01%); feed # 3 (control group without haematococcus pluvialis algae powder or haematococcus pluvialis algae residue added). Specific feed formulations and measured nutritional ingredients are shown in table 2.
TABLE 2 feed formulation and determination of nutrient composition
1.4 processing, packaging and preserving of river crab fattening feed
Various feed raw materials are subjected to superfine grinding (all raw materials pass through a 100-mesh screen), mixing and tempering to prepare sinking expanded feed, in order to prevent the oxidation loss of astaxanthin added in the feed, the expanded feed is granulated and then coated with grease on the feed by adopting a vacuum spraying process, so that an oxygen-isolating layer can be prevented from being formed, and the oxidation of the astaxanthin can be prevented. The particle size of the female sports fertilizer feed is about 5mm, and the length of the female sports fertilizer feed is about 8 mm; the expanded feed has good stability in water, and the dissolution rate of the expanded feed soaked in water for 4 hours (under the condition of 20 ℃) is less than 10%.
Because the crab fattening feed contains higher astaxanthin and highly unsaturated fatty acid, the crab fattening feed usually needs to be vacuumized and packaged, the outer layer is a dark color plastic woven bag to prevent the oxidative decomposition of the fatty acid and the astaxanthin caused by the feed after being exposed to light, the feed packaging bag is usually two layers, the inner layer is a vacuumized plastic bag, and the outer layer is a dark color woven bag; the feed is stored in a cool and dry place or a refrigerator with the temperature below-10 ℃.
1.5 female crab cultivation with physical fertilizer
The breeding experiment is carried out in an indoor circulating water system of a Chongming river crab research base of Shanghai ocean university, a breeding water tank is a PE (polyethylene) barrel (the diameter is × to 108 × cm), the water depth is kept at about 50cm during the experiment, continuous oxygenation is carried out for 24 hours, a fluorescent lamp (40W) is used as a light source, the light-dark period is 12 hours of illumination, the water source is fresh water after bleaching powder disinfection and aeration, 1# (algae residue), 2# (algae powder) and 3# (algae residue is not added) are respectively fed into three feed groups in the research, a certain number of female crabs are randomly placed into each water tank, each feed group is provided with 4 repeated water tanks, 12 experimental water tanks are obtained, experimental crabs are collected from the Chongming base of Shanghai ocean university, female crabs after shell removal are selected for a comparison experiment, the weight is 80g-90g, the average sex gland index (GSI) of the female crabs are 0.72% every other time, the experimental crabs are cultured in a temporary period of 3 days, the experiment, the experimental period is 3 days, the experimental period is 3, the water quality is adjusted, the average sex gland weight index of the water is adjusted, the water temperature of 0.5-15.7.7.7.7.7.7.10 mg, the total water, the water is measured, the water temperature of the total water is measured when the water is measured, the water temperature of the water is measured after the total water is measured, the total ammonia nitrogen intake is measured, the water is 1.7-20.7-10.7 days, the total time of the total ammonia nitrogen intake of the water, the water is measured, the water quality of the total water is measured after the total water is measured, the water is measured after the water is measured, the water is measured after the water is measured, the water is.
1.6 sampling and sample analysis.
After 60 days of experiment, the animals were stopped for 1 day, 2 females were randomly sampled from each PE bucket and weighed, and after dissection, gonads and livers were taken and GSI and HSI were calculated, and the samples were stored in a-40 ℃ freezer for subsequent analysis. The feed moisture (105 ℃ oven drying method), crude protein (Kjeldahl method) and crude ash (550 ℃ firing method) were measured according to the standard method of AOAC (1995); following the method of Folchet al (1957), chloroform: extracting the total lipids with methanol (V/V ═ 2:1) and measuring the content thereof; the carbohydrate content was determined by the 3, 5-dinitrosalicylic acid method (Zhang Jie, 1987). Fatty acid analysis the total lipids were methyl esterified using a 14% boron trifluoride-methanol solution, according to the method of Wu et al (2007). Sample preparation, antioxidant and immune index determination were performed according to the method of Zhao Lei et al (2016).
1.7. Evaluation of Effect
(1) Greatly reduces the cost of fattening feed
The natural astaxanthin from Haematococcus pluvialis powder is safe to use, has no toxic or side effect, but has high price, the current dry Haematococcus pluvialis powder (containing 3% astaxanthin) is about 1400 yuan/kg, and the price of the Haematococcus pluvialis residue is about 80 yuan/kg. The addition of 1% of haematococcus pluvialis residue and 0.2% of haematococcus pluvialis powder into the river crab fattening feed can ensure that the feed contains about 60mg/kg of astaxanthin, the content can meet the requirements of adult crab river crab coloring and quality regulation, and compared with the control group feed, the algae residue and the algae powder can respectively increase the feed cost by 800 yuan/ton and 2800 yuan/ton, but the two have no difference on the river crab fattening and coloring effects. Therefore, the haematococcus pluvialis residues are used as the astaxanthin source in the river crab fattening feed, so that the feed cost can be obviously reduced, and the reasonable utilization of the haematococcus pluvialis residues is facilitated.
TABLE 3 comparison of astaxanthin for three feeds and Total formulation cost
(2) Increasing survival rate and ensuring ovary development
The survival rate of the female after fattening can be improved to a small extent by adding haematococcus pluvialis residues into the feed, the 3# composition activity rate is 87%, and the 1# and 2# feed groups are 92.5% and 90% respectively. The addition of haematococcus pluvialis residues in the feed has no significant influence on the ovarian development of female river crabs, the fattening lasts for 60 days, the ovarian index reaches about 10 percent (figure 1), and the standard of first-class cream crabs is reached.
(3) Improving crab cream color and carotenoid content
As shown in figure 2, the color of the ovary dry sample of female crabs in the feed 1# (with added algae residue) group is not obviously different from that of the feed 2# (with added algae powder), the color of the feed 1# group is obviously better than that of the feed 3# group, the color values of all the formed crabs are shown in table 4, the ovary and cephalothorax a values of the feed 1# group and the feed 2# group are both obviously higher than that of the feed 3# group (P <0.05), the ovary L value and the ovary b value are the highest in the feed 3# group (P <0.05), and the hepatopancreas b value and cephalothorax L value of all the groups are not obviously different (P > 0.05).
The carotenoid composition in the three tissues of the female bodies is shown in Table 4. the contents of total carotenoid, astaxanthin, lutein and zeaxanthin in the ovaries of the female bodies of the feed 1# and the feed 2# groups are all obviously higher than that of the feed 3# group (P <0.05), and β -carotene content is the highest in the feed 3# group (P < 0.05). The contents of total carotenoid in the liver pancreas and total carotenoid and astaxanthin in the cephalospora are obviously higher than that of the feed 3# group (P <0.05) in the feed 1# and the feed 2# groups.
TABLE 4 Effect of the three feeds on the major carotenoid content in river crab ovary, hepatopancreas and crab shell (mg/kg dry tissue)
(4) Improving the antioxidant and immunity of female paste
Antioxidant indexes in sera of three groups of females and hepatopancreas are shown in Table 5. glutathione peroxidase (GSH-Px) and Catalase (CAT) in sera of the feed 1# and 2# groups are significantly higher than those of the feed 3# group (P <0.05), while MDA content is highest in the feed 3# group (P < 0.05). The hepatopancreas of the feed 3# group have NO significant difference in the rest of antioxidant indexes (P >0.05), except GR activity and MDA content, respectively, compared with those of the feed 1# and 2# groups (P < 0.05). The crab immunity indexes of each component are shown in Table 6. A L P activity, hemocyanin (Hc), and Nitric Oxide (NO) content in sera of the feed 1# and 2# groups are significantly higher than those of the feed 3# group (P <0.05), while ACP, A L P, γ -GT activity, and NO content are highest in the hepatopancreas (P < 0.05).
TABLE 5 Effect of three feeds on antioxidant indicators in serum and liver pancreas of female river crab
TABLE 6 influence of the three feeds on the immunological indices in the serum and in the liver pancreas of female river crabs
From the above embodiments, it can be seen that the innovation points of the present invention are mainly as follows: the haematococcus pluvialis residue after astaxanthin extraction not only contains conventional nutritional ingredients such as protein, fat and carbohydrate, but also contains a certain amount of astaxanthin, immune polysaccharide and other active substances, can improve the color and luster of crabs, and improve the immune disease resistance and nutritional quality, and therefore can be used as a high-grade feed raw material to perform high-value utilization on crab feed; secondly, because the demand of astaxanthin is different during the gonadal development period of the crab female and male individuals, and the demand of the male body on astaxanthin is far lower than that of the female body for the same species, two fattening feeds with different astaxanthin contents are prepared according to the different astaxanthin demands of the female and male individuals, so that the breeding cost can be saved, and the precise breeding can be realized; and thirdly, the crab fattening stage usually takes only 40-60 days, but has higher requirements on baits, and by feeding high-quality baits and scientific management, the gonad development can be promoted to improve the edible rate, and the nutritional quality of the crabs can be greatly improved, so that the haematococcus pluvialis residues have obvious using effect and higher added value in the crab fattening stage.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are given by way of illustration of the principles of the present invention, and that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (7)
1. A high-value utilization method of haematococcus pluvialis residues in economic crab fattening feed comprises the following steps:
(1) measuring the content of astaxanthin in the haematococcus pluvialis residues;
(2) determining the addition amount of haematococcus pluvialis residues in different economic crab fattening feed formulas,
wherein the addition amount of the haematococcus pluvialis residues in different economic crab fattening feed formulas is determined according to the demand amount of astaxanthin during gonadal development of female and male economic crabs and the astaxanthin content in the haematococcus pluvialis residues, the addition amount of the haematococcus pluvialis residues in the corresponding economic crab fattening feed formulas and other raw material formulas are calculated,
0.83% of haematococcus pluvialis residue needs to be added into the female crab fattening feed, and other raw material formulas in the female crab fattening feed are as follows: 10-15% of imported steam fish meal, 5-8% of meat meal, 18-24% of fermented soybean meal, 10% of peanut meal, 10-12% of rapeseed meal, 8-12% of corn, 14% of wheat middling, 4% of antarctic krill meal, 3% of soybean lecithin oil, 2.0% of refined fish oil, 2% of lard, 1% of soybean oil, 1% of rapeseed oil, 1% of monocalcium phosphate, 0.5% of choline chloride, 0.2% of vitamin complex, 0.3% of cholesterol, 0.5% of compound mineral, 0.2% of betaine and 50mg/kg of vitamin E;
(3) processing the economic crab fattening feed by using a suitable feed processing technology; and
(4) scientifically feeding the economic crabs with the economic crab fattening feed for fattening, and carrying out separate breeding on male and female economic crabs for fattening, wherein the ovary index of female crabs before fattening is 0.5-3%, and the ovary index after fattening is more than 7%, so that fattening can be stopped; the gonadal index before fattening the male crab is about 0.5%.
2. The high-value utilization method according to claim 1, wherein the economic crabs include eriocheir sinensis, portunus trituberculatus, scylla paramamosain, and blue crabs.
3. The high-valued utilization method according to claim 1 or 2, wherein the determination of the astaxanthin content in the Haematococcus pluvialis residue in step (1) comprises the steps of: extracting the Haematococcus pluvialis residue with acetone as solvent, and determining astaxanthin content by liquid chromatography.
4. The method for increasing the utilization rate of a liquid chromatography according to claim 3, wherein the column used in the liquid chromatography is L una3u Silica, and the mobile phase is n-hexane and acetone at a volume ratio of 83: 17.
5. The high-valued utilization method according to claim 1 or 2, wherein the suitable feed processing in step (3) includes sinking expanded feed processing and vacuum spraying of oil and fat to the surface of the feed to ensure the aquatic stability of the economic crab fattening feed and to prevent the oxidation loss of astaxanthin therein.
6. The high-value utilization method according to claim 5, wherein the suitable feed processing process in step (3) comprises the following steps:
(1) micronizing, mixing and modifying various feed raw materials to obtain sinking expanded feed, and granulating;
(2) spraying grease on the feed particles in vacuum;
(3) double-layer packaging, wherein the inner layer is a vacuumizable plastic bag, and the outer layer is a dark plastic woven bag;
(4) the feed is stored in dark place and stored in a cool and dry place or a refrigerator.
7. The high-value utilization method according to claim 1 or 2, wherein more than 80% of fed economic crabs fed in the step (4) have completed the molting of their genitals, and the feeding of the fattening feed for economic crabs can be started.
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