Method for Using Acremonium terricola Culture to Improve Growth and Immune Performance of Chinese Mitten Crabs

Abstract

The present invention discloses a method for using Acremonium terricola culture to improve the growth and immune performance of Chinese mitten crabs, belonging to the technical field of aquaculture. The Acremonium terricola culture comprises nutritional components with the following contents: ergosterol ≥ 550 mg / kg, adenosine ≥ 350 mg / kg. The indexes for characterizing the growth performance include: the survival rate, body weight, meat yield, crab paste index, etc. of Chinese mitten crabs. The indexes for characterizing the immune performance include: T-AOC, T-SOD, MDA, CAT in the muscle tissue of Chinese mitten crabs. After mixing the Acremonium terricola culture with the basal diet, the Chinese mitten crabs are fed; based on the dry weight of the basal diet, the addition amount of the Acremonium terricola culture is 0.15-0.3%. The present invention can significantly increase the edible part content, nutritional value, umami taste and sweetness of Chinese mitten crabs, thereby improving the meat quality and taste of Chinese mitten crabs, and can significantly improve the immune ability of Chinese mitten crabs.

Description

Technical Field

[0001] The present invention belongs to the technical field of aquaculture, and particularly relates to a method for using Acremonium terricola culture to improve the growth and immune performance of Chinese mitten crabs. Background Art

[0002] Chinese mitten crabs (Eriocheir sinensis) are not only delicious in meat but also rich in nutritional value, with relatively high aquaculture benefits. With the popularization of artificial feeding of Chinese mitten crabs, the main feeding method is using feed. Due to the insufficient content of nutritional components in the ingredients, the contents of various proteins and vitamins are relatively low, which easily causes the lack of proteins, fats, carbohydrates, minerals, vitamin powder, and trace elements in Chinese mitten crabs. Due to the lack of various nutrients, Chinese mitten crabs are prone to slow growth and development, small size, and various diseases.

[0003] Cordyceps gunnii is an endophytic fungus belonging to Ascomycota, Pyrenomycetes, Sphaeriales, and Clavicipitaceae, and parasitizes the larvae of Hepialidae. This fungus contains natural active substances such as cordycepin, cordycepic acid, and cordyceps polysaccharide. Acremonium terricola is a parasitic fungus of Cordyceps gunnii, and Acremonium terricola culture (ATC) is a feed additive obtained by fungal fermentation, which has functional components similar to those of Cordyceps gunnii. Research shows that supplementing Acremonium terricola culture can improve the growth performance of piglets and ducks, and enhance the antioxidant and immune functions of calves, and can effectively relieve the body's inflammatory response caused by mastitis in the treatment of rat mastitis. However, there is currently no research and report on the application of Acremonium terricola culture in improving the growth performance and immune ability of Chinese mitten crabs. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for using Acremonium terricola culture to improve the growth and immune performance of Chinese mitten crabs, so as to solve the following technical problems: how to effectively enhance the nutritional value of Chinese mitten crabs, improve the meat quality and taste, and at the same time improve the immune ability by using Acremonium terricola culture.

[0005] The purpose of the present invention can be achieved by the following technical solutions:

[0006] A method for using Acremonium terricola culture to improve the growth and immune performance of Chinese mitten crabs, wherein the Acremonium terricola culture comprises the following nutritional components in the following contents: ergosterol ≥ 550 mg / kg, adenosine ≥ 350 mg / kg.

[0007] As a further solution of the present invention, the indicators for characterizing the growth performance include: the survival rate, body weight, meat yield, crab paste index, percentage contents of protein and fat in crab meat and crab paste, percentage contents of triglyceride, free fatty acid, cholesterol, and phospholipid in crab meat and crab paste, and the content of free amino acids in crab meat.

[0008] As a further solution of the present invention, the indicators for characterizing the immune performance include: T-AOC (total antioxidant capacity), T-SOD (total superoxide dismutase activity), MDA (malondialdehyde content), and CAT (catalase activity) in the muscle tissue of Chinese mitten crabs.

[0009] As a further solution of the present invention, after mixing the acremonium terricola culture with the basal diet, the Chinese mitten crabs are fed; based on the dry weight of the basal diet, the addition amount of the acremonium terricola culture is 0.15-0.3%.

[0010] Beneficial effects of the present invention:

[0011] The present invention feeds Chinese mitten crabs with the acremonium terricola culture, which can significantly increase the edible part content, nutritional value, umami taste, and sweetness of Chinese mitten crabs, thereby improving the meat quality and taste of Chinese mitten crabs, and can significantly enhance the immune ability of Chinese mitten crabs. Specific embodiments

[0012] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0013] The acremonium terricola culture used in the embodiments of the present invention is produced by Hefei Microlife Biotechnology Co., Ltd. (batch number 20220103). After measurement, it contains ergosterol 632 mg / kg and adenosine 413 mg / kg.

[0014] Example 1

[0015] Method for using acremonium terricola culture to improve the growth and immune performance of Chinese mitten crabs

[0016] (1) The specific test design is as follows:

[0017] In the aquaculture ponds of a Chinese mitten crab aquaculture base in Anqing City, Anhui Province, plastic enclosures are used to divide the ponds into test ponds with equal size of 0.8 m 2 and 100 crab seedlings with a body weight of about 10 g are put in, and the male-female ratio is 1:1;

[0018] Three experimental ponds, namely the control group, experimental group 1, and experimental group 2, were set up, where:

[0019] Control group: 1.5 kg of basic feed was fed to every 100 kg of Chinese mitten crabs per day, without adding any supplementary feed and feed additives;

[0020] Experimental group 1: 1.5 kg of basic feed was fed to every 100 kg of Chinese mitten crabs per day, and 1.5 g (0.15%) of Acremonium terricola culture was added to each kilogram of basic feed.

[0021] Experimental group 2: 1.5 kg of basic feed was fed to every 100 kg of Chinese mitten crabs per day, and 3.0 g (0.3%) of Acremonium terricola culture was added to each kilogram of basic feed.

[0022] The three experimental groups were continuously fed for 210 days.

[0023] (2) After the breeding, data collection and index determination were carried out as follows:

[0024] (I) Data collection and index determination of growth performance:

[0025] ① Survival rate was statistically analyzed; 30 male and 30 female crabs were randomly selected from each experimental pond, weighed, and then dissected alive to separate crab meat and crab paste (in this application, the pancreas and gonads of Chinese mitten crabs are collectively referred to as crab paste), and the meat yield (meat yield = crab meat quality / crab body quality * 100%) and crab paste index (crab paste index = crab paste quality / crab body quality * 100%) were calculated. The results are shown in Table 1;

[0026] ② The percentage content of protein in crab meat or crab paste was determined by the Kjeldahl method;

[0027] The percentage content of fat in crab meat or crab paste: After weighing the crab meat or crab paste, it was vacuum-dried, then Soxhlet-extracted with anhydrous ether solvent, the solvent was removed, and then vacuum-dried and weighed again. The extract was the fat in the crab meat or crab paste. After weighing, the percentage content of fat in the crab meat or crab paste = the mass of fat in the crab meat or crab paste / the mass of the crab meat or crab paste * 100%. The results are shown in Table 2;

[0028] ③ The contents of triglyceride, free fatty acid, and phospholipid in crab meat or crab paste were determined by solid-phase extraction-gas chromatography; the contents of cholesterol in crab meat or crab paste were detected by solid-phase extraction-gas chromatography-mass spectrometry, and the percentage contents were further calculated; the results are shown in Table 3;

[0029] ④ Content of free amino acids in crab meat: Accurately weigh 2.50 g of crab meat, homogenize it with 20 mL of 5% trichloroacetic acid (TCA) solution, make up the volume to 25 mL, filter with qualitative filter paper, take 0.1 mL for centrifugation, perform pre-column derivatization reaction with OPA and FMOC derivatization reagents, and then analyze and detect with an amino acid-specific high-performance liquid chromatograph HP1100. The sample was repeatedly measured three times and the average value was taken. The results are shown in Table 4;

[0030] (II) Collection of immune performance data and determination of indicators:

[0031] Measure the T-AOC (total antioxidant capacity), T-SOD (total superoxide dismutase activity), MDA (malondialdehyde content), and CAT (catalase activity) of the crab leg muscle tissue respectively (according to the instructions of each corresponding kit of Nanjing Jiancheng Bioengineering Company), and the results are shown in Table 5;

[0032] Table 1 Effects of Acremonium terricola culture additive on the growth and development of Chinese mitten crabs

[0033]

[0034] Note: Different letters (a / b) indicate significant differences between groups (≤0.05)

[0035] It can be seen from Table 1 that in the experimental groups 1 and 2 with Acremonium terricola culture added to the basal diet, the survival rate and body weight of Chinese mitten crabs increased significantly. This result suggests that adding Acremonium terricola culture can significantly improve the aquaculture efficiency of Chinese mitten crabs; at the same time, the meat yield and crab paste index of experimental groups 1 and 2 also increased significantly, suggesting that adding Acremonium terricola culture can also significantly improve the nutritional value and edible quality of Chinese mitten crabs.

[0036] Table 2 Effects of Acremonium terricola culture additive on the biochemical composition of crab meat and crab paste of Chinese mitten crabs

[0037]

[0038] Note: Different letters (a / b) indicate significant differences between groups (≤0.05)

[0039] It can be seen from Table 2 that when feeding with Acremonium terricola culture added to the basal diet, the percentage content of protein in the crab meat and crab paste of Chinese mitten crabs increased significantly compared with those without adding Acremonium terricola culture, and the percentage content of fat remained basically unchanged compared with those without adding Acremonium terricola culture, further confirming that the nutritional value of Chinese mitten crabs in experimental groups 1 and 2 was significantly higher than that of the control group.

[0040] Table 3 Effects of Acremonium terricola culture additive on the lipid composition of crab meat and crab paste of Chinese mitten crabs

[0041]

[0042] Note: Different letters (a / b) indicate significant differences between groups (≤0.05). Unsaturated fatty acids / % refer to the percentage of unsaturated fatty acids in crab paste among free fatty acids.

[0043] As can be seen from Table 3, when feeding by adding Acremonium terricola culture to the basic feed, the percentage content of triglycerides in the crab meat and crab paste of Chinese mitten crabs increased significantly compared with those without adding Acremonium terricola culture. Lipids are important energy reserve substances for Chinese mitten crabs, and triglycerides are the main storage form, reflecting the nutritional value. Therefore, the energy accumulation in experimental group 1 and experimental group 2 is significantly better than that in the control group, and the nutritional value is significantly improved. Although the proportion of cholesterol in both crab meat and crab paste is not high, the percentage content of cholesterol in experimental group 1 and experimental group 2 still decreased significantly. N-3 and n-6 polyunsaturated fatty acids have extremely high nutritional value, especially for brain nutrition. The percentage content of n-3 and n-6 polyunsaturated fatty acids in the crab paste of experimental group 1 and experimental group 2 increased significantly. This result further confirms that adding Acremonium terricola culture significantly improves the nutritional value of Chinese mitten crabs.

[0044] Table 4 Effects of Acremonium terricola culture additive on flavor substances of Chinese mitten crabs

[0045] Index Control group Experimental group 1 Experimental group 2 Free amino acids (mg / g crab meat) <![CDATA[18.85±1.45 a > <![CDATA[22.78±1.89 b > <![CDATA[23.15±1.74 b > Arginine (mg / g crab meat) <![CDATA[6.53±0.22 a > <![CDATA[7.92±0.27 b > <![CDATA[7.83±0.28 b > Glycine (mg / g crab meat) <![CDATA[4.86±0.20 a > <![CDATA[5.06±0.10 a > <![CDATA[5.13±0.14 a > Alanine (mg / g crab meat) <![CDATA[3.58±0.33 a > <![CDATA[4.98±0.31 b > <![CDATA[5.10±0.40 b > Glutamic acid (mg / g crab meat) <![CDATA[0.51±0.04 a > <![CDATA[0.73±0.08 b > <![CDATA[0.701±0.07 b > Adenylic acid (mg / 100g crab meat) <![CDATA[64.20±5.15 a > <![CDATA[84.16±7.05 b > <![CDATA[81.55±8.035 b > Inosinic acid (mg / 100g crab meat) <![CDATA[33.30±3.04 a > <![CDATA[42.63±5.08 b > <![CDATA[45.63±4.704 b >

[0046] Note: Different letters (a / b) indicate significant differences between groups (≤0.05)

[0047] Arginine, glycine, and alanine in free amino acids are the main sources of the sweetness of crab meat; glutamic acid, adenosine monophosphate, and inosine monophosphate are the main sources of the umami of crab meat. As can be seen from Table 4, when feeding by adding Acremonium terricola culture to the basic feed, the contents of arginine, alanine, glutamic acid, adenosine monophosphate, and inosine monophosphate in the crab meat of Chinese mitten crabs increased significantly compared with those without adding Acremonium terricola culture. Therefore, the sweetness and umami of Chinese mitten crabs in experimental group 1 and experimental group 2 are significantly higher than those in the control group.

[0048] Table 5 Effects of Acremonium terricola culture additive on antioxidant capacity of Chinese mitten crab muscle

[0049] Group T-AOC (mmol / g) T-SOD (U / mg) CAT (U / mg) MDA (mmol / g) Control group <![CDATA[0.14±0.02 a > <![CDATA[81.45±5.14 a > <![CDATA[6.04±0.97 a > <![CDATA[0.12±0.04 a > Experimental group 1 <![CDATA[0.31±0.04 b > <![CDATA[123.47±8.11 b > <![CDATA[7.79±1.09 b > <![CDATA[0.13±0.03 a > Experimental group 2 <![CDATA[0.29±0.03 b > <![CDATA[131.84±7.75 b > <![CDATA[7.86±1.12 b > <![CDATA[0.11±0.04 a >

[0050] Feeding was carried out by adding Acremonium terricola culture to the basal diet. The T-AOC (total antioxidant capacity), T-SOD (total superoxide dismutase activity), and CAT (catalase activity) in the muscle tissue of Chinese mitten crabs were significantly increased compared with those without adding Acremonium terricola culture, and there was no obvious effect on MDA (malondialdehyde content). The immune response of shrimp and crab crustaceans mainly depends on non-specific immunity, and non-specific immune enzymes play an important role in the body's defense system. The antioxidant capacity of superoxide dismutase and catalase indirectly reflects the immune status of the animal body. Free radicals act on lipids to cause peroxidation reactions, and the end product of oxidation is malondialdehyde, which can cause cross-linking and polymerization of biological macromolecules such as proteins and nucleic acids and has cytotoxicity. Therefore, the immune ability of Chinese mitten crabs in experimental group 1 and experimental group 2 was significantly stronger than that of the control group.

[0051] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for using Acremonium terricola culture to improve the growth and immune performance of Chinese mitten crabs, characterized in that, The Acremonium terricola culture contains functional components in the following contents: ergosterol ≥ 550 mg / kg, adenosine ≥ 350 mg / kg.

2. The method for using the Acremonium terricola culture according to claim 1 to improve the growth and immune performance of Chinese mitten crabs, characterized in that, The indexes used to characterize the growth performance include at least one of the survival rate, body weight, meat yield, crab paste index, percentage contents of protein and fat in crab meat and crab paste, percentage contents of triglyceride, free fatty acid, cholesterol, and phospholipid in crab meat and crab paste, and the content of free amino acid in crab meat of Chinese mitten crabs.

3. The method for using the Acremonium terricola culture according to claim 1 to improve the growth and immune performance of Chinese mitten crabs, characterized in that, The indexes used to characterize the immune performance include at least one of T-AOC, T-SOD, MDA, and CAT in the muscle tissue of Chinese mitten crabs.

4. The method for using the Acremonium terricola culture according to claim 1 to improve the growth and immune performance of Chinese mitten crabs, characterized in that, After mixing the Acremonium terricola culture with the basal diet, the Chinese mitten crabs are fed; based on the dry weight of the basal diet, the addition amount of the Acremonium terricola culture is 0.15 - 0.3%.

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