CN115024436A - Feed capable of improving anti-stress capability of red-swamp crayfish in low-temperature environment - Google Patents
Feed capable of improving anti-stress capability of red-swamp crayfish in low-temperature environment Download PDFInfo
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- CN115024436A CN115024436A CN202210255565.9A CN202210255565A CN115024436A CN 115024436 A CN115024436 A CN 115024436A CN 202210255565 A CN202210255565 A CN 202210255565A CN 115024436 A CN115024436 A CN 115024436A
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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
-
- 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
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- 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
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- 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
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- 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
- A23K20/163—Sugars; Polysaccharides
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- 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
- A23K20/168—Steroids
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- 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
- A23K20/174—Vitamins
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- 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
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/30—Oligoelements
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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
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- 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)
- Proteomics, Peptides & Aminoacids (AREA)
- Birds (AREA)
- Marine Sciences & Fisheries (AREA)
- Insects & Arthropods (AREA)
- Inorganic Chemistry (AREA)
- Fodder In General (AREA)
Abstract
The invention relates to the technical field of aquaculture, in particular to a feed capable of improving the anti-stress capability of red-crayfish in a low-temperature environment. Specifically, the invention discloses application of a composition in preparing feed for improving the low-temperature resistance of red-shelled crayfish, wherein the composition comprises fish oil and soybean oil, and the mass ratio of the fish oil to the soybean oil is 3:1-1: 2.
Description
Technical Field
The invention relates to the technical field of aquaculture, in particular to a feed capable of improving the anti-stress capability of red-swamp crayfish in a low-temperature environment.
Background
The temperature is an important environmental factor influencing physiological and biochemical reactions of variable-temperature aquatic animals, and the water temperature change caused by seasonal changes is the most important reason influencing physiological and biochemical changes of aquatic animals. The change of the temperature has important influence on the feeding, growth, survival and the like of the aquatic animals, the changed environmental temperature can generate certain environmental pressure on the aquatic animals, in order to respond to the pressure, the aquatic animals can adjust the metabolic reaction of the aquatic animals to respond to the change, and when the adjustment of the aquatic animals is not enough to respond to the pressure caused by the environmental change, the aquatic animals can generate a series of pathological reactions and even die. The red-swamp crayfish is originally produced in Australia and introduced into China in 1992, and is tried to be cultured in the Guangdong Zhuhai and the Yangtze river delta area in China, so that the red-swamp crayfish has the advantages of being large in size, high in growth speed, strong in disease resistance and the like, and has a wide economic prospect. However, the red-crayfish, a bare-shell crayfish, has the physiological characteristic of being intolerant to low temperature because it is a warm-water-cultured shrimp.
Adverse environments such as low temperature, oxygen deficiency, nitrite stress and the like can cause stress response to aquatic animals, and further influence growth, survival, immunity and the like of the aquatic animals. The method for improving the immunity and disease resistance of aquatic animals by utilizing a nutriology method is an important way for realizing healthy culture, producing green aquatic products and ensuring the sustainable development of the aquaculture industry. At present, the research on nutrition and immunity of aquatic animals is more in fishes, and the research on shrimps and crabs is also gradually developed. More common immune supplements include antioxidants such as VC, VE, selenium, glutathione, and the like. Fatty acids, as one of the energy sources and nutrients essential to aquatic animals, have important biological and physiological regulatory functions in the body. Research has shown that fatty acids in the bait affect the anti-stress ability of fish under stress conditions. Fatty acid is also one of the main components constituting biological membrane, and the change of unsaturated fatty acid content in cell membrane can regulate the fluidity of cell membrane, and when the unsaturated fatty acid content in cell membrane is increased, the fluidity of cell membrane is also increased, so that the organism can maintain the change of cell membrane fluidity caused by environmental change of temperature, salinity, etc. by regulating the change of unsaturated fatty acid content in cell membrane. There have been some reports on the research on the lipid demand of the red-crayfish in the growing process, but the research on the nutritional demand of the red-crayfish in the adverse environment such as low temperature has not been reported.
Disclosure of Invention
The composition and the feed formula of the red-swamp crayfish effectively improve the low-temperature resistance of the red-swamp crayfish, such as the activity of antioxidant enzyme and the activity of immunoenzyme in hepatopancreata, hemolymph and/or gill tissues, and reduce the activity of transaminase, thereby reducing the adverse effect of low-temperature environment on the red-swamp crayfish.
In a first aspect of the invention, the application of a composition in preparing a feed for improving the low-temperature resistance of red-crayfish is provided, wherein the composition comprises fish oil and soybean oil, and the mass ratio of the fish oil to the soybean oil is 3:1-1: 2.
In another preferred example, the mass ratio of the fish oil to the soybean oil is 2: 1.
In another preferred embodiment, the improvement of the low temperature resistance includes: improving antioxidase activity, improving immunity enzyme activity, and reducing transaminase activity in blood lymph.
In another preferred example, increasing antioxidant enzyme activity comprises increasing SOD enzyme activity, increasing GPx enzyme activity, and/or increasing total antioxidant enzyme activity; and/or increasing antioxidant enzyme activity comprises increasing antioxidant enzyme activity in the hepatopancreas, hemolymph and/or gill tissue.
In another preferred embodiment, increasing the activity of the immunological enzyme comprises increasing the activity of AKP enzyme and/or the activity of ACP enzyme; and/or increasing the immunoenzyme activity comprises increasing the immunoenzyme activity in the hepatopancreas, hemolymph and/or gill tissues.
In another preferred embodiment, reducing transaminase activity comprises reducing GOP enzyme activity and/or GPT enzyme activity.
In another preferred example, the low temperature is 20 ℃ and below, preferably 15 ℃ and below, more preferably 9 ℃ and below.
In another preferred embodiment, the mass percentage of the composition in the feed is 2-10%, preferably 6%.
The invention also provides the use of a composition for improving the low temperature resistance of red-shelled crayfish, the composition comprising fish oil and soybean oil, and the mass ratio of the fish oil to the soybean oil is 3:1-1:2, preferably 2: 1.
The invention provides a feed for improving the low-temperature resistance of red-crayfish, which comprises the following components in percentage by mass: 40-50% of casein, 6-10% of gelatin, 2-10% of a composition of fish oil and soybean oil, 25-30% of corn starch, 2% of compound vitamin, 2% of compound mineral substance, 0-0.8% of betaine, 0.3-0.8% of choline chloride, 0-0.8% of cholesterol, 1-4% of carboxymethyl cellulose and the balance of alpha-cellulose, wherein the mass ratio of the fish oil to the soybean oil is 3:1-1: 2.
In another preferred embodiment, the components of the feed and the mass percentages of the components comprise: 40-50% of casein, 6-10% of gelatin, 2-10% of a composition of fish oil and soybean oil, 25-30% of corn starch, 2% of vitamin complex, 2% of compound mineral, 0-0.8% of betaine, 0.3-0.8% of choline chloride, 0-0.8% of cholesterol, 1-4% of carboxymethyl cellulose and the balance of alpha-cellulose, wherein the mass ratio of the fish oil to the soybean oil is 2: 1.
In another preferred example, the components of the feed and the mass percentages of the components comprise: 45% of casein, 8% of gelatin, 4% of fish oil, 2% of soybean oil, 26% of corn starch, 2% of compound vitamin, 2% of compound mineral, 0.5% of betaine, 0.5% of choline chloride, 0.5% of cholesterol, 2% of carboxymethyl cellulose and the balance of alpha-cellulose.
It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.
Drawings
FIG. 1 shows the effect of different high levels of unsaturated fatty acids on the resistance of the hepatopancreas of red-crayfish to oxidation at low temperatures. Wherein A is the activity change of the hepatopancreatic SOD enzyme; b is the change of the activity of the hepatopancreatic GPx enzyme; c is the change of the total antioxidant enzyme activity of the hepatopancreata; d is the content of liver pancreas MDA. 1-4 represent groups 1-4, respectively, i.e. the feed fed was 100% Fish Oil (FO), 2:1 fish oil to soybean oil ratio (SO33), 1:2 fish oil to soybean oil ratio (SO67) and 100% Soybean Oil (SO), respectively. Different letters represent significant differences (P < 0.05).
FIG. 2 shows the effect of different high unsaturated fatty acids on the hemolymph of red-crayfish against oxidation at low temperatures. Wherein A is the activity change of hemolymph SOD enzyme; b is the change of the hemolymph GPx enzyme activity; c is the change of the total antioxidase activity of hemolymph; d is the blood lymph MDA content. 1-4 represent groups 1-4, respectively, i.e. the feed fed was 100% Fish Oil (FO), 2:1 fish oil to soybean oil ratio (SO33), 1:2 fish oil to soybean oil ratio (SO67) and 100% Soybean Oil (SO), respectively. Different letters represent significant differences (P < 0.05).
FIG. 3 shows the effect of different high unsaturated fatty acids on the oxidation resistance of the gill tissue of the red-chelate crayfish at low temperature. Wherein A is the activity change of the branchial tissue SOD enzyme; b is the enzymatic activity change of the branchia tissue GPx; c is the change of the total antioxidant enzyme activity of the gill tissue; d is the content of the MDA in the gill tissue. 1-4 represent groups 1-4, respectively, i.e. the feed fed was 100% Fish Oil (FO), 2:1 fish oil to soybean oil ratio (SO33), 1:2 fish oil to soybean oil ratio (SO67) and 100% Soybean Oil (SO), respectively. Different letters represent significant differences (P < 0.05).
FIG. 4 is a graph showing the effect of varying levels of highly unsaturated fatty acids on the hepatopancreas alkaline phosphatase and acid phosphatase activities of red-camembered crayfish at low temperatures. Wherein A is the activity change of the hepatopancreas AKP enzyme; b is a change in hepatopancreatic ACP enzyme activity. 1-4 represent groups 1-4, respectively, i.e. the feed fed was 100% Fish Oil (FO), 2:1 fish oil to soybean oil ratio (SO33), 1:2 fish oil to soybean oil ratio (SO67) and 100% Soybean Oil (SO), respectively. Different letters represent significant differences (P < 0.05).
FIG. 5 is a graph showing the effect of varying levels of unsaturated fatty acids on the hemolymph alkaline phosphatase and acid phosphatase activities of red-camembelia crabae at low temperatures. Wherein A is the change in hemolymph AKP enzyme activity; b is a change in haemolymph ACP enzyme activity. 1-4 represent group 1-4, respectively, i.e., the feeds were fed with 100% Fish Oil (FO), 2:1 fish oil to soybean oil ratio (SO33), 1:2 fish oil to soybean oil ratio (SO67) and 100% Soybean Oil (SO). Different letters represent significant differences (P < 0.05).
FIG. 6 shows the effect of different high levels of unsaturated fatty acids on the activities of alkaline phosphatase and acid phosphatase in the gill tissue of red-crayfish at low temperatures. Wherein A is the enzymatic activity change of the branchial tissue AKP; and B is the activity change of the branchial tissue ACP enzyme. 1-4 represent groups 1-4, respectively, i.e. the feed fed was 100% Fish Oil (FO), 2:1 fish oil to soybean oil ratio (SO33), 1:2 fish oil to soybean oil ratio (SO67) and 100% Soybean Oil (SO), respectively. Different letters represent significant differences (P < 0.05).
FIG. 7 shows the effect of different high levels of unsaturated fatty acids on hemolymph transaminase activity of red-crayfish at low temperatures. Wherein A is GOP enzyme activity change in haemolymph; b is the GPT enzyme activity change in haemolymph. 1-4 represent groups 1-4, respectively, i.e. the feed fed was 100% Fish Oil (FO), 2:1 fish oil to soybean oil ratio (SO33), 1:2 fish oil to soybean oil ratio (SO67) and 100% Soybean Oil (SO), respectively. Different letters represent significant differences (P < 0.05).
Detailed Description
In order that the disclosure may be more readily understood, certain terms are first defined. As used in this application, each of the following terms shall have the meaning given below, unless explicitly specified otherwise herein. Other definitions are set forth throughout the application.
The term "about" can refer to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined.
The composition of the present invention refers to a composition comprising fish oil and soybean oil, and the mass ratio of the fish oil and the soybean oil is 3:1 to 1:2, preferably 2: 1.
The term "improved low temperature resistance" as used herein refers to an improvement in the low temperature stress resistance of a mariculture animal, preferably red crayfish.
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Experimental procedures without specific conditions noted in the following examples, generally followed by conventional conditions, such as Sambrook et al, molecular cloning: conditions described in a Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989), or according to the manufacturer's recommendations. Unless otherwise indicated, percentages and parts are by weight.
General procedure
1. Experimental feed
The experimental feed is a semi-purified feed, casein (Gansu Hualing casein Co., Ltd.) and gelatin (Chinese medicine reagent) are selected as protein sources, fish oil (Xiamen Xinsha medicine Co., Ltd.) and soybean oil (national Jinlongyu medicine Co., Ltd.) are selected as fat sources, corn starch (Kunshan Zhenleng food Co., Ltd.) is selected as sugar source, and a shrimp premix (Zhejiang Dengsheng Biotechnology Co., Ltd.) is selected as multi-mineral and multi-vitamin sources, and the specific components are shown in Table 1. The proportion of fish oil and soybean oil is respectively adjusted to prepare four feeds with different fatty acid contents, and the fat source components are respectively: 100% Fish Oil (FO), fish oil to soybean oil ratio of 2:1(SO33), fish oil to soybean oil ratio of 1:2(SO67), and 100% Soybean Oil (SO). Pulverizing all the raw materials, sieving with 80 mesh sieve, gradually amplifying, mixing, making into granulated feed with diameter of 3mm with meat grinder, air drying, and storing in refrigerator at-20 deg.C. The composition of the unsaturated fatty acids in each group of feeds is shown in table 2.
TABLE 1 feed Components in different experimental groups
a Provided by Zhejiang national Biotechnology Ltd
TABLE 2 composition of fatty acid content in different groups of feeds
a SFA:14:0,15:0,16:0,17:0,18:0,20:0.
b MUFA:16:1,17:1 18:1n-9,20:1n-9,22:1n-9,C24:1n-9.
c PUFA:18:2n-6,18:3n-3,20:2,22:2.
d HUFA:20:3n-6,20:5n-3,22:6n-3.
e n-3FA:18:3n-3,20:5n-3,22:6n-3.
f n-6FA:18:2n-6,20:3n-6.
2. Culture of experimental shrimps
The cultivation experiment is carried out in Jinshan Jing special aquaculture base, Shanghai city, the experimental shrimps are taken from a crayfish cultivation pond in the cultivation base, 400 crayfish with complete appendages and uniform size are selected for temporary cultivation, the water temperature is kept at 25 +/-1 ℃ during temporary cultivation, continuous oxygenation is carried out, the cultivation water is tap water which is aerated for more than 48 hours, the pH value is 7-8, and commercial feed is fed during the temporary cultivation. After one week of temporary culture, 320 high-activity crayfish are selected for culture experiments, 10 crayfish are placed in each culture box, the experiments are divided into 4 groups, each group is 8 in number, four feeds with different grease and acid resistance contents are respectively fed, the fat sources in the 4 groups of feeds respectively comprise 100% of Fish Oil (FO), the ratio of the fish oil to the soybean oil is 2:1(SO33), the ratio of the fish oil to the soybean oil is 1:2(SO67) and 100% of the Soybean Oil (SO), and the fed crayfish groups are respectively named group 1-4. The size of the cultivation box is 66 × 45 × 36cm, the initial weight is 34.63 ± 3.96g, and a PVC pipe is put in the box as a shelter and a place where the crayfish inhabits. Feeding twice every day at 8:00 am and 5:00 pm, wherein the feeding amount is about 3% of the shrimp weight, the rest cultivation conditions are the same as those in the temporary cultivation period, the water is changed after absorbing excrement and residual bait every day, the water change amount is 1/3 of the total water amount, and the feeding period is 4 weeks. And starting a low-temperature stress experiment after 4 weeks, dividing the shrimps in 4 feed groups into 4 temperature groups respectively, wherein one group is used as a control group, the other three groups are cooled at the speed of 1 ℃/12h to ensure that the final culture temperature is 25 +/-1 ℃, 20 +/-1 ℃, 15 +/-1 ℃ and natural low temperature respectively, each temperature group is repeated for two times, performing the temperature stress experiment for the periphery, the water temperature is controlled by a heating rod, the natural low-temperature group is not provided with the heating rod, and when the water temperature is lower than 7 ℃, properly heating and adding a foam box for heat preservation to ensure that the temperature is kept at 9 +/-2 ℃. The feeding mode of the feed is the same as that of the previous 4 weeks, the feeding amount is adjusted according to the food intake of each group of shrimps, and the fact that each group of shrimps are fed with full food is guaranteed, wherein the feed is not fed to the natural low-temperature group.
3. Collection of samples
After the 8-week experiment, starvation was performed for 24h, followed by weighing and counting. Three shrimps in the ecdysis interval are randomly selected in each jar, after ice bath anesthesia, hemolymph is extracted, the ratio of anticoagulant (3.52 g of sodium citrate, 8.42 of citric acid, 8.96g of NaCl, 0.74g of EDTA-2Na, 200ml of double distilled water and pH 7.3-7.4) to hemolymph is 1:1, one part of the anticoagulant is used for blood cell counting, the other part of the anticoagulant is centrifuged at 4000rpm at 4 ℃ for 15min, and blood cells and serum are separated for subsequent experiments. Respectively placing tissues of hepatopancreas, pancreas, stomach, intestine, muscle, etc. in a centrifuge tube, quickly placing in liquid nitrogen for quick freezing, and storing in a refrigerator at-80 deg.C for a long time. Survival and liver body index were calculated using the following formula:
survival (Survival,%) 100 × (shrimp tails at end/beginning of experiment);
liver body index (HIS,%) 100 × fresh liver weight/shrimp weight.
4. Determination of enzyme Activity
Weighing 0.1g of liver and pancreas tissues, adding sterile pre-cooled 0.86% physiological saline according to the ratio of 1:9, homogenizing in an ice bath, placing the sample in a refrigerated centrifuge for 15 minutes at 4 ℃, 3500rpm, and sucking the supernatant into a new centrifuge tube. All supernatants were assayed for protein concentration by Coomassie Brilliant blue followed by other enzyme activities. All enzyme activity determination kits are purchased from Nanjing institute of bioengineering, the specific determination method is described in the specification, and the enzyme activity of all tissue homogenates is determined within two weeks.
5. Data statistics and analysis
Data obtained from the experiment are expressed as Mean (Mean, M) ± standard deviation (Stdeva, SD), and the data obtained are statistically analyzed using SPSS 19 software. Differences of growth, survival and related enzyme activities of different feed feeding and different temperature groups are detected, a two-way ANOVA analysis method is adopted for significance test, and P <0.05 is used as a significance difference.
Example 1 Effect of adding different amounts of unsaturated fatty acids to feed on the antioxidant enzyme Activity of Procambrus cambrus at different temperatures
After four feeds with different lipid and acid-proof contents in the table 1 are fed for 4 weeks respectively, a low-temperature stress experiment is started, shrimps in the 4 feed groups are divided into 4 temperature groups respectively, one group is used as a control group, the other three groups are cooled at the speed of 1 ℃/12h, the final breeding temperature is 25 +/-1 ℃, 20 +/-1 ℃, 15 +/-1 ℃ and natural low temperature respectively, each temperature group is repeated for two times, and the temperature stress experiment for the periphery is carried out. The antioxidant enzyme activity and MDA content in hepatopancreas, hemolymph and branchial tissues of the red-swamp crayfish are detected after temperature stress.
1. The influence of adding unsaturated fatty acids with different contents in the feed on the antioxidant activity and MDA content in the hepatopancreas of the red-chelate crayfish under different temperatures
The change of the hepatopancreas SOD enzyme activity is shown in figure 1A, and the two-factor variance analysis result shows that the SOD enzyme activity is only influenced by the temperature change, the change of the content of highly unsaturated fatty acid does not have obvious influence on the SOD enzyme activity, the SOD enzyme activity in different groups shows a descending trend along with the reduction of the temperature, and the SOD enzyme activity at 9 ℃ is obviously reduced compared with that in other groups (P is less than 0.05).
The change of the activity of the hepatopancreas GPx enzyme is shown in figure 1B, and the result of the two-factor variance shows that the temperature change and the change of the content of highly unsaturated fatty acid both have obvious influence on the activity of the hepatopancreas GPx of the red-chelate photo-shell crayfish, and the activity of the GPx enzyme has a descending trend of different degrees along with the reduction of the temperature. Analysis of different highly unsaturated fatty acid addition groups at the same temperature shows that the activity of GPx enzyme in 100% fish oil group is obviously increased in the groups of 20 ℃, 15 ℃ and 9 ℃ compared with the 100% soybean oil group. For the 15 ℃ group and the 9 ℃ group, as the addition amount of the highly unsaturated fatty acid is higher, the activity of the hepatopancreatic GPx enzyme is higher, wherein the group1 (100% fish oil group) and the group2 (fish oil: soybean oil 2:1 group) are obviously higher than 100% soybean oil group.
The change of the total antioxidant enzyme activity of the hepatopancreas is shown in figure 1C, and the result of the two-factor variance shows that the temperature change and the change of the content of highly unsaturated fatty acid both have obvious influence on the total antioxidant enzyme activity of the hepatopancreas of the red-chelate crayfish, and the total antioxidant enzyme activity shows a gradual decline trend along with the reduction of the temperature. The analysis results of the addition levels of different highly unsaturated fatty acids at the same temperature showed that the hepatopancreas total antioxidant enzyme activity was significantly increased in group1 (100% fish oil group), group2 (fish oil: soybean oil 2:1 group) and group3 (fish oil: soybean oil 1:2 group) in the 20 ℃ group compared to the 100% soybean oil group. In the 15 ℃ group, the 100% fish oil group was significantly elevated compared to the remaining three added groups. In the 9 ℃ group, the total antioxidant enzyme activity of the hepatopancreas of 100 percent fish oil group is the highest, and the total antioxidant enzyme activity of each added group has no obvious difference.
The content of the MDA in the hepatopancreas is shown in fig. 1D, and the result of the two-factor anova shows that the content of the MDA in the hepatopancreas is mainly affected by temperature change, but is not significantly affected by different contents of the highly unsaturated fatty acids. With decreasing temperature, the MDA content in the hepatopancreas gradually increased, with a significant increase in MDA content in 9 ℃ compared to the remaining groups.
TABLE 3 temperature and highly unsaturated fatty acid content analysis of oxidative two-factor variance of hepatopancreas of red-chelate Callicarpa nudiflora
2. The influence of adding unsaturated fatty acids with different contents in the feed on the antioxidant activity and MDA content in hemolymph of the red-camembelia crayfish at different temperatures
SOD enzyme activity in haemolymph is shown in figure 2A, and two-factor anova results show that SOD enzyme activity in haemolymph is affected by both temperature change and the addition amount of highly unsaturated fatty acid. Wherein SOD enzyme activity of 100% fish oil addition groups in each temperature group is not obviously changed, and the SOD enzyme activity of fish oil: the soybean oil 2:1 added group showed no significant difference between the groups except for the decrease in enzyme activity at 20 ℃. Fish oil: the SOD enzyme activity is lowest in the group with soybean oil added in a ratio of 1:2 and at the temperature of 9 ℃, and the difference between the other groups is not obvious. In 100% of soybean oil, except for the group at 25 ℃, the SOD enzyme activity of each of the other groups was significantly reduced. The SOD enzyme activities in the groups except 25 ℃ are not obviously changed, and the SOD enzyme activities in the rest temperature groups are gradually reduced along with the reduction of the addition amount of the fish oil. In particular 15 ℃ and 9 ℃ groups, 100% addition group, fish oil: the SOD enzyme activity in the haemolymph of the soybean oil 2:1 addition group is obviously higher than that of the fish oil: soybean oil 1:2 additive group and 100% soybean oil group.
The change of GPx activity in haemolymph is shown in figure 2B, and the result of two-factor anova shows that the GPx activity in haemolymph is only affected by the change of temperature, and the influence of the addition amount of highly unsaturated fatty acid on the GPx activity is small. The GPx enzyme activity was lowest in each of the added groups at 9 ℃, significantly lower than in the remaining groups (P < 0.05).
The change of the total antioxidant enzyme activity in haemolymph is shown in fig. 2C, and the result of two-factor anova shows that the temperature and the addition amount of highly unsaturated fatty acid both have significant influence on the change of the total antioxidant enzyme activity, and the change trend is similar to the change trend of the total antioxidant enzyme activity in hepatopancreas, namely, the total antioxidant enzyme activity is in a whole descending trend along with the reduction of the temperature and the reduction of the addition amount of fish oil.
The MDA content in haemolymph is shown in fig. 2D, and the results of two-factor anova analysis show that the MDA content in haemolymph is only related to the temperature change, that is, the MDA content gradually increases with the decrease of temperature, and the MDA content in each group is the highest in the group at 9 ℃, and is significantly higher than that in the other groups.
TABLE 4 temperature and highly unsaturated fatty acid content analysis of oxidative two-factor variance of hemolymph of Procambrus clarkii
3. The influence of adding unsaturated fatty acids with different contents in the feed on the antioxidant activity and MDA content in the branchia of the red-swamp crayfish under different temperatures
The SOD enzyme activity in the gill tissue is shown in figure 3A, and the two-factor anova result shows that the SOD enzyme activity in the gill tissue is only related to the temperature change, namely the SOD enzyme activity in the gill tissue is in a descending trend along with the reduction of the temperature in general, and the SOD enzyme activity in all the other groups except for group3 (fish oil: soybean oil 1:2) is found to be remarkably reduced compared with the control group when the temperature is 9 ℃.
GPx enzyme activity in gill tissues is shown in fig. 3B, and two-way anova results show that GPx enzyme activity is only related to temperature changes, independent of unsaturated fatty acid addition, and that GPx enzyme activity is significantly reduced in each temperature group compared to the control group (P < 0.05).
The change of the total antioxidant enzyme activity in gill tissues is shown in fig. 3C, and the results of two-factor anova show that the total antioxidant enzyme activity in gill tissues is affected by both the temperature change and the change of the addition amount of highly unsaturated fatty acids. The effect of temperature on this was similar to the trend of change in hepatopancreas and haemolymph, i.e. the total antioxidant enzyme activity decreased in the hypothermic group. In the 20 ℃ group, the change of the total antioxidant enzyme activity in each addition group is not obviously different. In the 15 ℃ group, the total antioxidant enzyme activity gradually decreases with the decrease of the addition amount of the fish oil, and the activity in 100% of fish oil (group 1) is the highest and is obviously higher than that in 100% of soybean oil addition group.
The MDA content in gill tissue is shown in fig. 3D, and the two-factor anova results show that there is no significant difference between the MDA accumulation amount in gill tissue and the addition amount of unsaturated fatty acid, and only the temperature change is related, and in all nutrition groups, the MDA content is significantly increased in the lowest temperature group compared with the control group (P < 0.05).
TABLE 5 temperature and highly unsaturated fatty acid content analysis of the antioxidant two-factor variance of the gill tissue of red-chela
Example 2 Effect of adding different amounts of unsaturated fatty acids to feed on Immunity activity of Red Tiger crayfish at different temperatures
The experimental procedure is as in example 1 and the activity of the immunoenzymes of the hepatopancreas, hemolymph and gill tissues of the red-swamp crayfish is examined after temperature stress.
1. Influence of adding different unsaturated fatty acids in feed on activity of immunoenzyme in hepatopancreas of red-swamp crayfish at different temperatures
The activity of AKP enzyme in the hepatopancreas is shown in fig. 4A, and the results of two-way anova show that the activity of AKP enzyme is affected only by temperature change, and gradually decreases with decreasing temperature, and all of them show a significant decrease in the 9 ℃ group.
The ACP enzyme activity in the hepatopancreas is shown in FIG. 4B, and the result of two-factor anova shows that the ACP enzyme activity is simultaneously influenced by temperature change and high unsaturated fatty acid content change, and the ACP enzyme activity and the high unsaturated fatty acid content change have certain interaction, wherein the ACP enzyme activity in the 9 ℃ group is the lowest and is obviously lower than that in the rest temperature groups, and the ACP enzyme activity in the 100% fish oil addition group is wholly higher than that in the rest addition groups.
TABLE 6 temperature and highly unsaturated fatty acid content analysis of variance of two factors on the hepatopancreas alkaline and acid phosphatase activities of red-camembelia crabae
2. Influence of adding different content of unsaturated fatty acid in feed on activity of immunoenzyme in hemolymph of red-chelia crayfish at different temperatures
The AKP enzyme activity in the haemolymph is shown in figure 5A, and the two-factor analysis of variance shows that the AKP enzyme activity in the haemolymph is influenced by temperature change and the addition amount of highly unsaturated fatty acid at the same time, and the AKP enzyme activity shows a gradual reduction trend along with the reduction of the temperature, wherein the AKP enzyme activity in the 25 ℃ group is the highest and is obviously higher than that in other temperature groups, while the AKP enzyme activity in the 9 ℃ group is obviously lower than that in other temperature groups, and the comparison of the AKP enzyme activity in different highly unsaturated fatty acid addition groups shows that the AKP enzyme activity in group2 (fish oil: soybean oil 2:1) is obviously higher than that in other addition groups at the same temperature.
ACP enzyme activity in haemolymph as shown in FIG. 5B, two-way anova showed that ACP enzyme activity in haemolymph was affected only by temperature change, with ACP enzyme activity being highest in the 25 ℃ group and lowest in the 9 ℃ group at the lowest temperature, except for group2 (fish oil: soybean oil 2: 1).
TABLE 7 temperature and polyunsaturated fatty acid content two-way ANOVA on Red-crayfish hemolymph alkaline phosphatase and acid phosphatase Activity
3. Influence of adding unsaturated fatty acids with different contents in feed on activity of immunoenzymes in branchia of red-swamp crayfish at different temperatures
The AKP enzyme activity in gill tissue is shown in fig. 6A, and the results of two-factor anova show that the AKP enzyme activity in gill tissue is affected by both temperature change and the addition amount of highly unsaturated fatty acid, and in different highly unsaturated fatty acid addition groups, the AKP enzyme activity is highest in the 25 ℃ group and lowest in the 9 ℃ group. Under the temperature condition of 15 ℃, the addition amount of the fish oil is gradually reduced. AKP enzyme activity was lowest in 100% of soybean oil groups in the 9 ℃ group.
ACP enzyme activity in gill tissues is shown in fig. 6B, and two-way anova results show that ACP enzyme activity in gill tissues is only affected by temperature changes, and ACP enzyme activity gradually decreases with decreasing temperature, wherein ACP enzyme activity in each addition group is highest in the 25 ℃ group and lowest in the 9 ℃ group.
TABLE 8 temperature and high unsaturated fatty acid content analysis of variance of two factors on the activity of alkaline phosphatase and acid phosphatase in the gill tissue of Procambrus cambruensis
Example 3 Effect of adding different amounts of unsaturated fatty acids to feed on the Activity of Valeriana camara and Glutamine-alanine transferase in Helinba of Red Tiger Callicarpa at different temperatures
The experimental method is the same as that in example 1, and the activity of the glutamic acid and glutamic-pyruvic transaminase in the bloody lymph of the red-chela crayfish is detected after the temperature stress.
GOP activity in haemolymph is shown in figure 7A, and the two-factor anova result shows that the GOP enzyme activity in haemolymph is only affected by temperature change, and the GOP enzyme activity in haemolymph gradually increases with the decrease of temperature, wherein the GOP enzyme activity in the 15 ℃ group and the 9 ℃ group is obviously increased compared with the control group.
GPT activity in hemolymph is shown in fig. 7B, and the results of two-factor anova showed that GPT enzyme activity in hemolymph was affected by both temperature change and the amount of highly unsaturated fatty acid added, and similar to GOP enzyme activity change, both showed a gradual increase tendency with increasing temperature, and by comparing GPT enzyme activity in different unsaturated fatty acid added groups, it was found that there was no significant difference in GOP activity between groups in the 25 ℃ group, GPT enzyme activity was the lowest in 100% fish oil group in the 20 ℃ group and 15 ℃ group, and GPT enzyme activity was the lowest in two mixed oil groups when the temperature was decreased to 9 ℃ group, which was significantly higher than that in the fish oil group and soybean oil group.
TABLE 9 temperature and highly unsaturated fatty acid content analysis of variance of two factors on hemolymph transaminase activity of red-camembelia crayfish
Discussion of the related Art
The low temperature inhibits the growth, metabolism, nonspecific immunity and the like of the red-camembelia crayfish, the food intake is gradually reduced along with the reduction of the temperature, the food intake is even stopped in the lowest temperature group, and the metabolism of the organism is maintained by consuming the nutrient substances stored by the body, so that the research on the metabolic demand of the red-camembelia crayfish at the low temperature and the nutrition enhancement of the red-camembelia crayfish before the red-camembelia crayfish enters the overwintering environment have important significance for helping the red-camembelia crayfish to live through the adverse environmental conditions such as the low temperature.
According to the invention, soybean oil and fish oil are selected as fat sources, and researches on the requirement of the fat source of the red-crayfish have been reported, and the optimal growth effect of the red-crayfish is obtained when the soybean oil is used as the fat source under the optimal temperature, probably because the high unsaturated fatty acid content in the soybean oil can meet the survival requirement of the red-crayfish in a warm water breeding environment.
Through previous researches, the activity of the antioxidant enzyme of the red-chelate crayfish is reduced along with the reduction of the temperature, so that excessive ROS in a body cannot be effectively eliminated, and the oxidative damage of the body is caused. The invention also discovers that the activities of the antioxidase in different feed addition groups are in a descending trend along with the reduction of the temperature, and simultaneously discovers that the activities of the antioxidase in three tissues are influenced by the addition amount of n-3/n-6 fatty acid besides the influence of the temperature. The activity of the two immune enzymes has a similar trend to that of antioxidant enzymes, i.e. the different feed additive groups are gradually reduced with the temperature. It is shown that under low temperature conditions, both the antioxidant and immunoenzymatic activities of red-crayfish were inhibited, similar to previous studies. Meanwhile, the activity of antioxidant enzyme and immune enzyme is also influenced by the addition amount of n-3/n-6 fatty acid in the feed. The detection of the oxidation injury marker MDA and the tissue injury marker enzyme glutamic-pyruvic transaminase finds that the content change of the MDA is only influenced by the temperature change, and the change of the content of the unsaturated fatty acid has little influence on the change. However, the activity of glutamic-pyruvic transaminase in hemolymph is influenced by temperature change and the addition amount of n-3/n-6 fatty acid in feed, the activity of GPT enzyme is the lowest in 100% fish oil group in the 20 ℃ group and the 15 ℃ group, and the activity of GPT enzyme in the two mixed oil groups is the lowest when the temperature is reduced to 9 ℃, which is obviously lower than that of the fish oil group and soybean oil group, probably because the oxidation damage in the 9 ℃ group is serious, while the content of unsaturated fatty acid in the fish oil is increased, the accumulation of unsaturated fatty acid in cell membranes is promoted, the risk of cell oxidation damage is increased, and further serious tissue damage is caused.
In general, the invention finds that along with the increase of the addition amount of n-3/n-6 fatty acid in the feed, the activity of antioxidant in tissues of the red-crayfish as well as the degree of oxidative damage is reduced, and indicates that n-3 unsaturated fatty acid has a certain promotion effect on the resistance of the red-crayfish to low-temperature stress, but the excessive addition amount of fish oil can easily increase the lipid peroxidation risk of cell membranes, so the invention considers that the ratio of the n-3/n-6 fatty acid in the fish oil: the soybean oil is added at a ratio of 2:1, and the effect is best.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.
Claims (10)
1. Use of a composition in the preparation of a feed for improving the low temperature resistance of red-shelled crayfish, wherein the composition comprises fish oil and soybean oil, and the mass ratio of the fish oil to the soybean oil is 3:1-1: 2.
2. The use according to claim 1, wherein the ratio of fish oil to soybean oil is 2:1 by mass.
3. The use of claim 1, wherein said improving low temperature resistance comprises: improving antioxidase activity, improving immunity, and reducing transaminase activity in haemolymph.
4. The use of claim 3, wherein increasing antioxidant enzyme activity comprises increasing SOD enzyme activity, increasing GPx enzyme activity, and/or increasing total antioxidant enzyme activity; and/or increasing antioxidant enzyme activity comprises increasing antioxidant enzyme activity in the hepatopancreas, hemolymph and/or gill tissues.
5. The use of claim 3, wherein increasing the activity of an immunoenzyme comprises increasing the activity of AKP enzyme and/or ACP enzyme; and/or increasing the immunoenzyme activity comprises increasing the immunoenzyme activity in the hepatopancreas, hemolymph and/or gill tissues.
6. The use of claim 3, wherein reducing transaminase activity comprises reducing GOP enzyme activity and/or GPT enzyme activity.
7. Use according to claim 1, wherein the composition is present in the feed in an amount of 2-10%, preferably 6% by weight.
8. The feed for improving the low-temperature resistance of the red-crayfish, which is characterized by comprising the following components in percentage by mass: 40-50% of casein, 6-10% of gelatin, 2-10% of a composition of fish oil and soybean oil, 25-30% of corn starch, 2% of vitamin complex, 2% of compound mineral, 0-0.8% of betaine, 0.3-0.8% of choline chloride, 0-0.8% of cholesterol, 1-4% of carboxymethyl cellulose and the balance of alpha-cellulose, wherein the mass ratio of the fish oil to the soybean oil is 3:1-1: 2.
9. The feed according to claim 7, wherein the feed comprises the following components in percentage by mass: 40-50% of casein, 6-10% of gelatin, 2-10% of a composition of fish oil and soybean oil, 25-30% of corn starch, 2% of vitamin complex, 2% of compound mineral, 0-0.8% of betaine, 0.3-0.8% of choline chloride, 0-0.8% of cholesterol, 1-4% of carboxymethyl cellulose and the balance of alpha-cellulose, wherein the mass ratio of the fish oil to the soybean oil is 2: 1.
10. The feed as claimed in claim 7, wherein the components of the feed and the mass percentages of the components comprise: 45% of casein, 8% of gelatin, 4% of fish oil, 2% of soybean oil, 26% of corn starch, 2% of compound vitamin, 2% of compound mineral, 0.5% of betaine, 0.5% of choline chloride, 0.5% of cholesterol, 2% of carboxymethyl cellulose and the balance of alpha-cellulose.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102379376A (en) * | 2010-09-02 | 2012-03-21 | 华东师范大学 | Forage capable of improving broodstock reproductive performance of Cherax quadricarionatus |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102379376A (en) * | 2010-09-02 | 2012-03-21 | 华东师范大学 | Forage capable of improving broodstock reproductive performance of Cherax quadricarionatus |
Non-Patent Citations (1)
| Title |
|---|
| 吴东蕾: "低温胁迫和脂质营养对红螯光壳螯虾生长及生理的影响", 中国博士学位论文全文数据库农业科技辑, no. 8, pages 052 - 9 * |
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