CN113940314B - Method for improving composition of black soldier fly body PUFAs by using linseed oil and detection method thereof - Google Patents
Method for improving composition of black soldier fly body PUFAs by using linseed oil and detection method thereof Download PDFInfo
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- CN113940314B CN113940314B CN202111157816.1A CN202111157816A CN113940314B CN 113940314 B CN113940314 B CN 113940314B CN 202111157816 A CN202111157816 A CN 202111157816A CN 113940314 B CN113940314 B CN 113940314B
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- 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
- A23K20/105—Aliphatic or alicyclic compounds
-
- 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
-
- 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
- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Environmental Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Birds (AREA)
- Insects & Arthropods (AREA)
- Animal Behavior & Ethology (AREA)
- Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses a method for improving the constitution of black soldier fly body PUFAs by using linseed oil, which comprises the following steps: a disposable injection molding meal box is used as a black soldier fly larva feeding container, and a vent is cut out of the middle of the meal box cover by scissors so that the black soldier fly larva breathes; adding the black soldier fly larvae and the prepared daily ration into a disposable injection molding cutlery box, putting the cutlery box into a biochemical incubator, keeping the temperature at 28 ℃, ventilating and supplementing water every day, wherein the supplementing water amount is based on the condition that the humidity of the daily ration is 70%, adding the daily ration for multiple times, and adding the daily ration by taking the remaining 50% of the last feeding as a reference; taking the first blackened larva appearing in the disposable injection molding cutlery box as a signal, and collecting all the first blackened larva at one time. Based on the current situation, the method for changing PUFAs in the black soldier fly larvae by using the linseed oil is innovatively developed to fully exert the excellent characteristics of the black soldier fly, and the n-6/n-3 ratio is reduced. Therefore, the invention can remarkably improve the economic value of the hermetia illucens.
Description
Technical Field
The invention belongs to the technical field of animal feeds, and particularly relates to a method for improving the constitution of hermetia illucens PUFAs by using linseed oil and a detection method.
Background
Essential fatty acids refer to fatty acids which are essential for maintaining the body's function, but which are not synthesized by the body and must be supplied by food. There are two types of well-defined human essential fatty acids, one is n-3 series polyunsaturated fatty acids based on alpha-Linolenic acid (ALA); the other is n-6 series polyunsaturated fatty acid (Polyunsaturated fatty acid, PUFAs) with Linoleic Acid (LA) as matrix. Along with the continuous and deep research of unsaturated fatty acid, n-3 series polyunsaturated fatty acid is found to play an important role in promoting normal development of brain and vision formation in the processes of stress and tissue function recovery after wound operation, improving immunity, inhibiting inflammatory reaction, promoting growth and development, improving lipid metabolism, preventing and treating cardiovascular and cerebrovascular diseases, regulating and controlling organism hormone metabolism and a plurality of enzyme activities. The ratio of n-6PUFAs to n-3PUFAs in diet is related to cardiovascular diseases, and reducing the ratio has the effect of preventing and treating cardiovascular diseases.
However, most of the residents in China eat mainly corn oil, rapeseed oil and soybean oil with higher LA content in daily life, so that the dietary LA of the residents in China is seriously out of standard, ALA is seriously insufficient, the ratio of n-6PUFAs to n-3PUFAs in the diet of the residents is far higher than the recommended ratio of 4:1-6:1 of n-6PUFAs to n-3PUFAs in the diet of the people, and the ratio of n-6PUFAs to n-3PUFAs in the human body is generally higher. Therefore, increasing the content of n-3PUFAs in a food product is of great importance for providing a healthy diet for humans. Land animals have little DHA and EPA in the body due to the intake of docosahexaenoic acid (Docosahexaenoic acid, DHA) and eicosapentaenoic acid (Eicosapentaenoic acid, EPA), so that the DHA and EPA content in common fish bodies is very small, and the DHA content in deep sea fish in cold areas, such as salmon, sardine and the like, is very high, so that the deep sea fish oil is a traditional dietary source for providing n-3 polyunsaturated fatty acids. Because of the large difficulty and small quantity of deep sea fish fishing, the cost of supplementing n-3 polyunsaturated fatty acid by the deep sea fish oil is high, and the problems of fish taste, smell, toxin content, allergy and the like are not completely solved, the application of the deep sea fish oil is still limited. Flaxseed is one of the most abundant sources of plant ALA and can be used as a substitute for deep sea fish oil. Linseed oil has a higher ALA content of more than 50% of total fatty acids and a relatively lower LA content of about 13% of total fatty acids compared to other edible oils. Intake of ALA has been considered to have a positive impact on the prevention of cardiovascular disease based on the results of clinical trials, epidemiological surveys and experimental studies. Because of the high ALA content of flaxseeds, the use of flaxseeds against cardiovascular diseases has been advocated.
The black soldier fly is an important saprophytic insect with the functions of resource regeneration and environmental protection, and has the advantages of large biomass, high food conversion efficiency and high propagation speed. The black soldier fly has the characteristic of wide ingestion range, and various organic wastes (such as kitchen waste, livestock manure, spoiled vegetables and fruits, crop wastes and food processing wastes) can be utilized by the black soldier fly and converted into various nutritional ingredients such as protein, lipid, peptides, amino acids, chitin, vitamins, phospholipid and the like. The transformed hermetia illucens somatic proteins and amino acids have been used to produce aquaculture feeds and feeds with high digestibility. The black soldier fly as an economic insect for treating wastes has higher utilization value, and researches show that the content of crude protein in the black soldier fly larvae reaches more than 44% -48%, the content of crude fat reaches 25% -30%, and the black soldier fly larvae has higher nutritive value, can be used as a good feed raw material and has higher utilization value. Normally grown hermetia illucens larvae in nature have a n-6/n-3PUFAs ratio of about 18:1, if the n-6/n-3PUFAs ratio can be reduced to be below 4:1, the economic utilization value can be obviously improved. However, in the current development process of the hermetia illucens industry, most of researches mainly focus on simple technical aspects of feeding, worm processing and the like, and researches on how to better improve the utilization value of the hermetia illucens are relatively few. Therefore, the method for reducing the n-6/n-3PUFAs ratio in the hermetia illucens larvae has great significance for improving the economic value of the hermetia illucens and the industrial development of the hermetia illucens.
By searching, no patent publication related to the present patent application has been found.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a method for improving the constitution of black soldier fly body PUFAs by using linseed oil and a detection method.
The technical scheme adopted for solving the technical problems is as follows:
a method for improving the constitution of black soldier fly body PUFAs by using linseed oil comprises the following steps:
a disposable injection molding meal box is used as a black soldier fly larva feeding container, and a vent is cut out of the middle of the meal box cover by scissors so that the black soldier fly larva breathes;
adding the black soldier fly larvae and the prepared daily ration into a disposable injection molding cutlery box, putting the cutlery box into a biochemical incubator, keeping the temperature at 28 ℃, ventilating and supplementing water every day, wherein the supplementing water amount is based on the condition that the humidity of the daily ration is 70%, adding the daily ration for multiple times, and adding the daily ration by taking the remaining 50% of the last feeding as a reference;
taking the first blackened larva appearing in the disposable injection-molded cutlery box as a signal, and collecting all the blackened larva at one time to obtain the compound feed;
wherein, the composition components and the mass portions of the prepared daily ration are as follows:
3.0 parts of soybean oil, 2.0 parts of linseed oil, 93.68 parts of chicken manure with water content of 70%, 0.01 part of tertiary butyl hydroquinone and 2% tween-201.32 parts; the fraction is a mass fraction;
the preparation method comprises the following steps: mixing soybean oil, linseed oil and tertiary butyl hydroquinone for standby, adding 2% tween-20 into chicken manure with water content of 70%, fully mixing, finally adding a mixture consisting of soybean oil, linseed oil and tertiary butyl hydroquinone, and fully stirring to obtain the chicken manure.
Further, the hermetia illucens larvae are hatched by the following method:
taking rice bran and wheat bran according to the mass ratio of 1:1, adding into a culture box, adding water to adjust the humidity to 80%, uniformly dispersing hermetia illucens eggs on the surface of a substrate, putting into a biochemical incubator, and keeping the temperature at 28 ℃; daily observation is carried out until macroscopic larvae appear on the surface of the substrate, and hatching is completed.
Further, the disposable injection molding cutlery box is made of transparent materials.
Further, the disposable injection molding cutlery box is 650ml, and the vent is 12cm 6cm.
Further, when the prepared ration is prepared, the edible oil and the tertiary butyl hydroquinone are mixed by a seed method, and the mixing method is as follows:
the tertiary butyl hydroquinone is completely dissolved in a small amount of edible oil to prepare 10% tertiary butyl hydroquinone grease solution, and then the tertiary butyl hydroquinone is directly added into the residual grease to be uniformly stirred.
The detection method of PUFAs composition of the hermetia illucens body obtained by the method comprises the following steps:
collecting the obtained hermetia illucens bodies, and cleaning the residual feces on the surfaces of the hermetia illucens bodies by utilizing the peristaltic characteristic points of the larvae; subjecting the insect body to 24-hour hunger treatment to discharge undigested residues in the digestive tract of the insect body;
removing primary water from the harvested larvae in an electrothermal blowing drying oven at 60-70 ℃ to prepare a semi-dry sample and break the semi-dry sample; the above operations are carried out in an environment with the temperature of 18-25 ℃ and the air humidity of less than 25% except for the special description;
determining the fatty acid according to standard GB5009.168-2016 of an internal standard method in determination of fatty acid in food safety national standards; the sample pretreatment method adopts a hydrolysis-extraction method.
The invention has the advantages and positive effects that:
1. based on the current situation, the method for changing PUFAs in the black soldier fly larvae by using the linseed oil is innovatively developed to fully exert the excellent characteristics of the black soldier fly, and the n-6/n-3 ratio is reduced. Therefore, the invention can remarkably improve the economic value of the hermetia illucens.
2. The method for improving the black soldier fly body PUFAs by using the linseed oil is characterized in that soybean oil, tertiary butyl hydroquinone, linseed oil and tween-20 are reasonably combined, so that the fat nutrition concentration in the feed and the mixing uniformity of all components can be improved, and the problems of excessively rapid oxidation and deterioration of the feed and the like are prevented.
3. According to the method, the black soldier fly is raised in a manner of adding the linseed oil, wherein the content of n-3PUFAs of the black soldier fly larvae can be obviously improved by adding the linseed oil with the concentration of 2%, and the n-6/n-3 ratio is reduced. The method for improving the constitution of the hermetia illucens body PUFAs by using the linseed oil is suitable for culturing the hermetia illucens larvae, and lays a foundation for changing the constitution of the hermetia illucens body PUFAs, improving the economic value of the hermetia illucens and transforming, popularizing and applying.
4. The feed for improving the method formed by the hermetia illucens PUFAs in the method comprises the following components: linseed oil: flax is an annual herb, linseed oil is rich in ALA, accounting for more than 50% of total fatty acids, while LA is relatively low, accounting for about 13% of total fatty acids. According to research reports, ALA can prevent cardiovascular diseases, inhibit inflammatory response and improve immunity.
5. The culturing box for raising the hermetia illucens, which is used for improving the constitution of the hermetia illucens PUFAs, is used as a culturing box of a culturing container for the hermetia illucens larva, and a vent of 12cm x 6cm is cut out of the middle of the box cover by scissors, so that the culturing space can be greatly saved, the raising cost is reduced, and the design of the box cover not only prevents the larva from escaping, but also enhances the ventilation of the larva. The transparent material is convenient for observing the activity condition of the hermetia illucens in real time.
6. The method for improving the composition of the hermetia illucens PUFAs in the method can ensure the normal growth of the hermetia illucens and reduce the adverse effect of high temperature on the hermetia illucens larvae as much as possible. The humidity of the method can keep the skin of the body surface of the hermetia illucens moist, and is favorable for peristalsis foraging.
7. According to the method, the daily ration adding frequency of the method for improving the PUFAs of the hermetia illucens bodies can be improved, so that the hermetia illucens can be promoted to eat, and the accumulation and fermentation of daily ration and the breeding of fly maggots are avoided.
Drawings
FIG. 1 shows the standard chromatographic peak separation patterns of 37 fatty acids in the present invention.
Detailed Description
The following describes the embodiments of the present invention in detail, but the present embodiments are illustrative and not limitative, and are not intended to limit the scope of the present invention.
The raw materials used in the invention are conventional commercial products unless specified; the methods used in the present invention are conventional in the art unless otherwise specified.
A method for improving the constitution of black soldier fly body PUFAs by using linseed oil comprises the following steps:
a disposable injection molding meal box is used as a black soldier fly larva feeding container, and a vent is cut out of the middle of the meal box cover by scissors so that the black soldier fly larva breathes;
adding the black soldier fly larvae and the prepared daily ration into a disposable injection molding cutlery box, putting the cutlery box into a biochemical incubator, keeping the temperature at 28 ℃, ventilating and supplementing water every day, wherein the supplementing water amount is based on the condition that the humidity of the daily ration is 70%, adding the daily ration for multiple times, and adding the daily ration by taking the remaining 50% of the last feeding as a reference;
taking the first blackened larva appearing in the disposable injection-molded cutlery box as a signal, and collecting all the blackened larva at one time to obtain the compound feed;
wherein, the composition components and the mass portions of the prepared daily ration are as follows:
3.0 parts of soybean oil, 2.0 parts of linseed oil, 93.68 parts of chicken manure with water content of 70%, 0.01 part of tertiary butyl hydroquinone and 2% tween-201.32 parts; the fraction is a mass fraction;
the preparation method comprises the following steps: mixing soybean oil, linseed oil and tertiary butyl hydroquinone for standby, adding 2% tween-20 into chicken manure with water content of 70%, fully mixing, finally adding a mixture consisting of soybean oil, linseed oil and tertiary butyl hydroquinone, and fully stirring to obtain the chicken manure.
Preferably, the hermetia illucens larvae are hatched by the following method:
taking rice bran and wheat bran according to the mass ratio of 1:1, adding into a culture box, adding water to adjust the humidity to 80%, uniformly dispersing hermetia illucens eggs on the surface of a substrate, putting into a biochemical incubator, and keeping the temperature at 28 ℃; daily observation is carried out until macroscopic larvae appear on the surface of the substrate, and hatching is completed.
Preferably, the disposable injection molding cutlery box is made of transparent materials.
Preferably, the disposable injection-molded cutlery box is 650ml, and the ventilation opening is 12cm by 6cm.
Preferably, when the prepared ration is prepared, the edible oil and the tertiary butyl hydroquinone are mixed by a seed method, and the mixing method is as follows:
the tertiary butyl hydroquinone is completely dissolved in a small amount of edible oil to prepare 10% tertiary butyl hydroquinone grease solution, and then the tertiary butyl hydroquinone is directly added into the residual grease to be uniformly stirred.
The detection method of PUFAs composition of the hermetia illucens body obtained by the method comprises the following steps:
collecting the obtained hermetia illucens bodies, and cleaning the residual feces on the surfaces of the hermetia illucens bodies by utilizing the peristaltic characteristic points of the larvae; subjecting the insect body to 24-hour hunger treatment to discharge undigested residues in the digestive tract of the insect body;
removing primary water from the harvested larvae in an electrothermal blowing drying oven at 60-70 ℃ to prepare a semi-dry sample and break the semi-dry sample; the above operations are carried out in an environment with the temperature of 18-25 ℃ and the air humidity of less than 25% except for the special description;
determining the fatty acid according to standard GB5009.168-2016 of an internal standard method in determination of fatty acid in food safety national standards; the sample pretreatment method adopts a hydrolysis-extraction method.
Specifically, the relevant preparation and detection examples are as follows:
examples 1-3 methods for improving the composition of hermetia illucens PUFAs Using linseed oil
Examples 1 to 3 are the composition of the daily ration, the feeding conditions, the harvesting time and the treatment method, respectively, of the method for improving the composition of the hermetia illucens PUFAs by using linseed oil, and the composition of the raw materials for preparing the daily ration are shown in Table 1.
TABLE 1 examples 1-3 ration configurations
Adding 500g of rice bran and 500g of wheat bran into a culture box, adding water to regulate the humidity to about 80%, uniformly dispersing ovum on the surface of a substrate, placing into a biochemical incubator, and keeping the temperature at 28 ℃. Daily observation is carried out until macroscopic larvae appear on the surface of the substrate, and hatching is completed.
A disposable injection molding cutlery box is used as a black soldier fly larva feeding container, and 72cm of the middle of the cutlery box cover is cut out by scissors 2 The ventilation openings are used for enabling the hermetia illucens larvae to breathe. Each group is provided with 4 repeats, 200 larvae are counted in each repeat, the larvae and the prepared daily ration are added into a disposable injection molding cutlery box together, the disposable injection molding cutlery box is placed into a biochemical incubator to be kept at 28 ℃, ventilation and water supplement are carried out every day, the water supplement amount is based on the condition that the humidity of the daily ration is about 70%, the daily ration is added for multiple times, the rest about 50% of the last feeding is used as a reference, and the specific adding time and the adding amount are as shown in tables 2-4.
TABLE 2 first feed addition (day 0 of feeding)
TABLE 3 second feed addition (day 4 of feeding)
TABLE 4 third feed addition (day 10 of feeding)
Note that: the first blackened larva appearing in the disposable injection-molded cutlery box is taken as a signal, the first blackened larva is collected at one time, and the residual excrement on the surface of the first blackened larva is cleaned by utilizing the peristaltic characteristic point of the larva. It was starved for 24 hours to remove undigested residues in the digestive tract of the insects. Then removing primary water in an electrothermal blowing drying oven at 60-70deg.C to obtain semi-dry sample, grinding into powder with a mortar, and performing fat extraction (see Table 5) and fatty acid profile analysis (see tables 6 and 7).
The correlation detection of the invention is as follows:
verification test of method for improving black soldier fly body PUFAs composition by using linseed oil:
(1) Test animal selection and grouping
With the adoption of a completely random design of a single-factor experimental scheme, 2400 1-year-old hermetia illucens larvae with similar hundred worm weights and strong peristaltic capacities are selected and randomly divided into 3 groups (example 1, example 2 and example 3), each group is divided into 4 repeats, 200 larvae are repeated, fresh chicken manure added with 5% of soybean oil is used as a control group (example 1), and fresh chicken manure added with linseed oil to replace 2% (example 2) and 5% (example 3) of the soybean oil is used as an experimental group respectively.
(2) Feed and feeding management
The control group is fed with the feed of the example 1, the test group is fed with the test feed which is added with linseed oil to replace the feed for improving the growth performance of the black soldier fly by 2 percent (example 2) and 5 percent (example 3), and the test group is fed with the test feed for improving the growth performance of the black soldier fly by adding linseed oil, and the test group is fed with the test feed freely in the test process. A650 ml disposable injection-molded cutlery box is used as a black soldier fly larva feeding container, and a 12cm x 6cm vent is cut out of the middle of the cutlery box cover by scissors so that the black soldier fly larva breathes. Adding 1-year larva and feed for improving the growth performance of hermetia illucens into a disposable injection molding cutlery box, placing the cutlery box into a biochemical incubator, keeping the temperature at 28 ℃, ventilating and supplementing water every day, wherein the water supplementing amount is based on the condition that the feed humidity is about 70%, adding the feed for multiple times, and adding the feed by taking the rest about 50% of the last feeding as a reference.
(3) Fatty acid determination step and method
The first blackened larva appearing in the disposable injection-molded cutlery box is taken as a signal, the first blackened larva is collected at one time, and the residual excrement on the surface of the first blackened larva is cleaned by utilizing the peristaltic characteristic point of the larva. It was starved for 24 hours to remove undigested residues in the digestive tract of the insects. The primary water was removed in an electrothermal forced air drying oven at 60-70 c to prepare a semi-dry sample, and the fatty acid determination was performed as follows. The fatty acid was measured according to the standard of the internal standard method (GB 5009.168-2016) in the measurement of fatty acids in foods in food safety national standards. The sample pretreatment method adopts a hydrolysis-extraction method, petroleum ether is used as a leaching solvent to carry out fat leaching in a full-automatic fat analyzer, 150mg of extracted worm fat is accurately weighed by an analytical balance, and 2ml of an internal standard solution of undecanoic acid triglyceride (C11:0) is added. Adding 2% NaOH-CH3OH solution for saponification, using 15% BF3-CH3OH solution for fatty acid methyl esterification, and filling the prepared sample liquid into a sample bottle for measurement by an on-machine. The set gas chromatography conditions and methods are as follows: the capillary chromatographic column is DB-WAX capillary chromatographic column (Agilent, 30m×250 μm×0.25 μm); the temperature of the sample inlet is 250 ℃; the detector temperature was 280 ℃; the initial temperature of the column box is 40 ℃ after the temperature programming for 2min; heating at 40-171 deg.c and 45 deg.c/min for 25min; heating at 171-215 deg.C at 10 deg.C/min for 20min; the carrier gas is helium; the split ratio is 10:1, the sample injection volume is 3 mu L, and the gas flow is 2mL/min. And then separating out standard chromatographic peaks by using 37 fatty acid methyl ester mixed standard substances, and determining the peak outlet time and the retention time of each chromatographic peak, wherein the separation condition of the standard chromatographic peaks is shown in figure 1.
(4) Test results
TABLE 5 determination of content of coarse fat of hermetia illucens larvae
As can be seen from table 5, the crude fat content in example 2 and example 3 was significantly higher in hermetia illucens larvae than in example 1 (27.53%, 31.34% vs 25.50%) (P < 0.05); the difference in crude fat content was not significant compared to example 3 in example 2.
TABLE 6 hermetia illucens ration fatty acid composition
Note that: ND is undetected, different lowercase letters indicate significance of the difference (P < 0.05), unlabeled letters or the same letters indicate no significance of the difference (P > 0.05). The content units of the rest fatty acids except n-6/n-3 are g/100g of the sample.
TABLE 7 fatty acid composition of hermetia illucens bodies
Note that: ND is undetected, different lower case letters indicate difference significance (P < 0.05), (P < 0.01) representing extreme significance. The unlabeled letter or the same letter indicates that the difference is not significant. The content units of the rest fatty acids except n-6/n-3 are g/100g of the sample.
As can be seen from Table 7, the larval content n-3PUFAs level of example 3 is significantly higher than that of example 2 and example 1 (P < 0.05), and the larval n-3PUFAs level of example 2 is significantly higher than that of example 1 compared with example 1. The differences between the content groups in the larval fatty acids were very significant (P < 0.01), and the levels of example 1, example 2n-6PUFAs were significantly higher than in example 3 (P < 0.05). There was no significant difference in n-6PUFAs levels between example 1 and example 2 (P < 0.05). Examples 1, 2 and 3, the n-6/n-3 ratios were 12.877, 2.688 and 0.851, respectively.
In view of economic cost, example 2 was recommended as an optimal method for improving the constitution of hermetia illucens PUFAs.
Although embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the disclosure of the embodiments.
Claims (1)
1. A method for improving the constitution of black soldier fly body PUFAs by using linseed oil is characterized in that: the method comprises the following steps:
a disposable injection molding meal box is used as a black soldier fly larva feeding container, and a vent is cut out of the middle of the meal box cover by scissors so that the black soldier fly larva breathes;
adding the black soldier fly larvae and the prepared daily ration into a disposable injection molding cutlery box, putting the cutlery box into a biochemical incubator, keeping the temperature at 28 ℃, ventilating and supplementing water every day, wherein the supplementing water amount is based on the condition that the humidity of the daily ration is 70%, adding the daily ration for multiple times, and adding the daily ration by taking the remaining 50% of the last feeding as a reference; the specific adding time and the adding amount are as follows:
the first time of feeding on day 0, the feed is added with the following amount:
the feed is added for the second time on the 4 th day of feeding, and the feed addition amount is as follows:
the third feed is added on the 10 th day of feeding, and the feed addition amount is as follows:
taking the first blackened larva appearing in the disposable injection-molded cutlery box as a signal, and collecting all the blackened larva at one time to obtain the compound feed;
wherein, the composition components and the mass portions of the prepared daily ration are as follows:
3.0 parts of soybean oil, 2.0 parts of linseed oil, 93.68 parts of chicken manure with water content of 70%, 0.01 part of tertiary butyl hydroquinone and 1.32 parts of 2% tween-20; the fraction is a mass fraction;
the preparation method comprises the following steps: mixing soybean oil, linseed oil and tertiary butyl hydroquinone for standby, adding 2% tween-20 into chicken manure with water content of 70%, fully mixing, finally adding a mixture consisting of soybean oil, linseed oil and tertiary butyl hydroquinone, and fully stirring to obtain the chicken manure;
the hermetia illucens larvae are hatched by the following method:
taking rice bran and wheat bran according to the mass ratio of 1:1, adding into a culture box, adding water to adjust the humidity to 80%, uniformly dispersing hermetia illucens eggs on the surface of a substrate, putting into a biochemical incubator, and keeping the temperature at 28 ℃; daily observation is carried out until macroscopic larvae appear on the surface of the matrix, and hatching is completed;
the disposable injection molding cutlery box is made of transparent materials;
the disposable injection molding cutlery box is 650ml, and the ventilation opening is 12cm x 6cm;
when the prepared ration is prepared, the edible oil and the tertiary butyl hydroquinone are mixed by a seed method, and the mixing method is as follows:
the tertiary butyl hydroquinone is completely dissolved in a small amount of edible oil to prepare 10% tertiary butyl hydroquinone grease solution, and then the tertiary butyl hydroquinone is directly added into the residual grease to be uniformly stirred;
the detection method for the PUFAs composition of the hermetia illucens bodies obtained by the method comprises the following steps:
collecting the obtained hermetia illucens bodies, and cleaning the residual feces on the surfaces of the hermetia illucens bodies by utilizing the peristaltic characteristic points of the larvae; subjecting the insect body to 24-hour hunger treatment to discharge undigested residues in the digestive tract of the insect body;
removing primary water from the harvested larvae in an electrothermal blowing drying oven at 60-70 ℃ to prepare a semi-dry sample and break the semi-dry sample; the above operations are carried out in an environment with the temperature of 18-25 ℃ and the air humidity of less than 25% except for the special description;
determining the fatty acid according to standard GB5009.168-2016 of an internal standard method in determination of fatty acid in food safety national standards; the sample pretreatment method adopts a hydrolysis-extraction method;
the ratio of n-6/n-3 was 2.688.
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