Radix pseudostellariae microecological preparation as well as preparation method and application thereof
Preparing the microecological preparation of ginseng through specific extraction and fermentation methods has solved the problem that the effective ingredients of ginseng are difficult to extract and absorb, and the growth performance and disease resistance of ginseng are improved.
Patent Information
- Application Number
- CN202510146665.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-10
- Publication Date
- 2025-06-03
AI Technical Summary
In the prior art, the active ingredients of Prince Ginseng are difficult to be extracted and absorbed, resulting in unsatisfactory effects in the breeding of yellow croaker and waste of resources.
A specific method is used to prepare the root extract of Prince Ginseng and ferment and culture with probiotics to form a microecological preparation of Prince Ginseng. Through specific extraction and fermentation steps, this preparation improves the precipitation of nutrients and active substances such as genus ginseng polysaccharides and saponins.
Through the synergistic effect, the growth performance and disease resistance of yellow croaker are improved, the biological activity of digestive enzymes is enhanced, and the feed utilization and survival rate are improved.
Smart Images
Figure CN120078099A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to a large yellow croaker feed additive, in particular to a Pseudo-radix pseudoginseng microecological preparation and a preparation method and application thereof. Background Art
[0002] With the expansion of the scale of large yellow croaker farming, the increase in farming density and the fact that it mainly uses fresh fish as bait, the problems of frequent diseases and slow growth are becoming increasingly prominent. The traditional method of using antibiotics and chemical drugs to prevent and control fish diseases has drug resistance and toxic side effects, which seriously endangers the safety and health of farmed animals and humans. Therefore, the development and use of environmentally friendly feed additives that can promote the growth of farmed aquatic animals and enhance their disease resistance is a new challenge facing the aquaculture industry.
[0003] Pseudostellariae Pseudostellariae has the characteristics of being pure natural, free of drug residues, non-resistant and with little toxic side effects. It can be used as a feed additive in large yellow croaker farming. However, Pseudostellariae Pseudostellariae is a plant-derived medicinal material. Its effective ingredients are relatively complex and mostly exist in the cytoplasm of cells. The plant cell wall structure is densely composed of cellulose, hemicellulose, etc. The active ingredients of traditional Chinese medicine are difficult to extract, the absorption and utilization rate is low, the effect is not ideal, and resources are wasted. Summary of the invention
[0004] The purpose of the present invention is to provide a Pseudostellaria heterophylla microecological preparation and a preparation method and application thereof.
[0005] In order to solve the problems existing in the prior art, the technical solution adopted by the present invention is:
[0006] In a first aspect, the present invention provides a method for preparing a probiotic preparation of Radix Pseudostellariae, comprising the following steps:
[0007] (1) Preparation of Pseudostellariae Radix Extract:
[0008] Wash and air-dry the root tubers of Pseudostellaria heterophylla, grind them into powder, pass through a 50-70 mesh sieve, and place the powder in a round-bottom flask, add 75-85% ethanol, reflux extract in a water bath at 90±1℃ for 2-4h, filter while hot, extract the residue with 95±1℃ ultrapure water for 3-5h and filter, mix and concentrate the two filtrates, remove water, and obtain Pseudostellaria heterophylla extract, which is stored in a refrigerator at 4±1℃ for later use;
[0009] (2) The Pseudostellariae Radix extract was fermented in a probiotic shaker for 23-25 hours to prepare the Pseudostellariae Radix microecological preparation, which was then placed in a refrigerator at 4±1°C for later use.
[0010] After the root extract of Pseudostellaria chinensis is extracted by the above-mentioned specific method, it is conducive to the precipitation of nutrients and active substances such as Pseudostellaria chinensis polysaccharides and saponins, and then combined with the step of probiotic shaker fermentation culture, the nutritional and medicinal value of Pseudostellaria chinensis can be maximized, thereby promoting growth. The inventor has found through many experiments that if the extraction method of Pseudostellaria chinensis is changed to water extraction, the growth performance of large yellow croaker cannot be significantly improved.
[0011] Furthermore, the concentration of the Pseudostellariae Radix Extract is 35-45 mg / mL, preferably 40 mg / mL. If the concentration of the Pseudostellariae Radix Extract is less than 35 mg / mL, the precipitation of nutrients and active substances such as Pseudostellariae Polysaccharides and Saponins cannot be improved, and if the concentration of the Pseudostellariae Radix Extract is greater than 45 mg / mL, the effect does not change significantly with the increase of concentration.
[0012] Furthermore, the inoculation amount of probiotics is 4-6%, preferably 5%.
[0013] Furthermore, the probiotics are Candida utilis, Bacillus subtilis or plant lactic acid bacteria.
[0014] Furthermore, the Candida utilis is cultured at 30±1° C. and 190-210 r / min.
[0015] After a large number of experiments, the inventors found that the Pseudostellariae Radix extract prepared in step (1) and Candida utilis are best cultured at 30±1°C and 190-210r / min, and the precipitation of nutrients and active substances such as Pseudostellariae Radix polysaccharides and saponins can be maximized.
[0016] Furthermore, the Bacillus subtilis and plant lactic acid bacteria are cultured at 37±1°C and 170-190r / min.
[0017] After a large number of experiments, the inventors found that when the Pseudostellariae Radix Extract prepared in step (1) is fermented in a shaking table with Bacillus subtilis or plant lactic acid bacteria, the best effect can be achieved only when it is cultured at 37±1°C and 170-190r / min, and the precipitation of nutrients and active substances such as Pseudostellariae Radix polysaccharides and saponins can be maximized.
[0018] In a second aspect, the present invention provides a Pseudostellaria heterophylla probiotic preparation, which is prepared by the preparation method described in the first aspect.
[0019] In a third aspect, the present invention provides the use of the Pseudostellaria heterophylla microecological preparation described in the second aspect as a feed additive for large yellow croaker.
[0020] Fourthly, the present invention provides a large yellow croaker feed, which comprises the Pseudostellaria heterophylla microecological preparation described in the second aspect above.
[0021] Furthermore, the addition amount of the Pseudostellaria heterophylla microecological preparation is 0.4-0.6% of the feed quality.
[0022] The advantages and beneficial effects of the present invention are as follows:
[0023] After the specific probiotics in the Pseudostellaria heterophylla microecological preparation of the present invention and the Pseudostellaria heterophylla extract are combined in a specific ratio according to a specific method, the two can play a synergistic effect, which can improve the precipitation of nutritional components and active substances such as Pseudostellaria heterophylla polysaccharides and saponins, is conducive to accelerating the absorption of large yellow croakers, and maximally exerts the nutritional and medicinal value of Pseudostellaria heterophylla, thereby promoting growth; it can also produce various digestive enzymes and antibacterial substances, which can improve the biological activity of digestive enzymes in the digestive tract, enabling large yellow croakers to better digest and absorb nutrients in the gastrointestinal tract, improve feed utilization rate, and enhance disease resistance, thereby improving the survival rate of large yellow croakers, and has a significant effect on improving the growth performance of large yellow croakers. Description of the Drawings
[0024] Figure 1 Effect of the Pseudostellaria heterophylla microecological preparation on the survival rate of large yellow croakers; Figure 1 Different letters indicate significant significant differences (P<0.05)
[0025] Figure 2 Effect of the Pseudostellaria heterophylla microecological preparation on the growth performance of large yellow croakers; Figure 2 Different letters indicate significant significant differences (P<0.05); in the figure: A is the absolute weight gain rate; B is the absolute growth rate;
[0026] Figure 3 Effect of the Pseudostellaria heterophylla microecological preparation on the serum antioxidant enzyme activity of large yellow croakers; Figure 3 Different letters indicate significant significant differences (P<0.05); in the figure, A is the serum SOD activity, B is the serum CAT activity, and C is the serum GSH-Px activity;
[0027] Figure 4 Effect of the Pseudostellaria heterophylla microecological preparation on the non-specific immune enzyme activity of large yellow croakers; Figure 4 Different letters indicate significant significant differences (P<0.05), in the figure, A is the serum ACP activity, B is the serum AKP activity, and C is the serum LZM activity. Detailed Embodiments
[0028] The following will clearly describe the technical solutions in the embodiments of the present application in conjunction with the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art belong to the scope of protection of the present application.
[0029] In the following experimental methods of the embodiments, unless otherwise specified, they are all conventional methods, carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following embodiments, unless otherwise specified, can all be obtained from commercial channels.
[0030] Example 1
[0031] This example provides a Pseudostellaria heterophylla microecological preparation, and its preparation method includes the following steps:
[0032] (1) Prepare the Pseudostellaria heterophylla extract:
[0033] ① Wash and air-dry the Pseudostellaria heterophylla tuberous roots, grind them into powder, and pass through a 60-mesh sieve to obtain Pseudostellaria heterophylla tuberous root powder;
[0034] ② Place 100 g of Pseudostellaria heterophylla tuberous root powder in a round-bottom flask, add an ethanol solution with a volume concentration of 80%, and the addition amount is 10 times the weight-volume ratio. Reflux and extract at 90 °C in a water bath for 3 h;
[0035] ③ Filter while it is hot, and extract the residue with ultrapure water at 95 °C with a weight-volume ratio of about 10 times for 4 h and then filter;
[0036] ④ Mix and concentrate the two filtrates, remove the water, and obtain a Pseudostellaria heterophylla extract with a concentration of 40 mg / mL, and store it in a refrigerator at 4 °C for later use.
[0037] ⑤ Ferment and culture the Pseudostellaria heterophylla extract with Candida utilis in a shaker at 30 °C and 200 r / min for 24 h to prepare the Pseudostellaria heterophylla microecological preparation, and store it in a refrigerator at 4 °C for later use. The inoculation amount of Candida utilis is 5%.
[0038] Example 2
[0039] This example provides a Pseudostellaria heterophylla microecological preparation. The difference between this example and Example 1 is only that in step ⑤, the Pseudostellaria heterophylla extract is fermented and cultured with Bacillus subtilis in a shaker at 37 °C and 180 r / min for 24 h to prepare the Pseudostellaria heterophylla microecological preparation, and the rest are the same as in Example 1.
[0040] Example 3
[0041] This example provides a Pseudostellaria heterophylla microecological preparation. The difference between this example and Example 1 is only that in step ⑤, the Pseudostellaria heterophylla extract is made into a Pseudostellaria heterophylla microecological preparation by shaking flask fermentation culture with plant lactic acid bacteria at 37 °C and 180 r / min for 24 h, and the rest is the same as in Example 1.
[0042] Comparative Example 1
[0043] This comparative example provides a Pseudostellaria heterophylla microecological preparation. The difference between this comparative example and Example 1 is only that step ⑤ is missing, and the rest is the same as in Example 1.
[0044] Experimental Example 1
[0045] 1 Test materials
[0046] 1.1 Test fish
[0047] Large yellow croakers were purchased from Ningde Dingcheng Aquatic Products Co., Ltd., and juvenile fish with complete body shape, good health and vitality, an average body length of (7.87 ± 0.68) cm and a body weight of (7.77 ± 1.99) g were selected.
[0048] 1.2 The Pseudostellaria heterophylla microecological preparations fermented by Candida utilis, Bacillus subtilis and plant lactic acid bacteria were respectively prepared by the method described in Example 1.
[0049] 1.3 Preparation of test feed
[0050] The artificial compound feed is the commercially available "Haima" brand large yellow croaker feed, and the nutritional components are: crude fat ≥ 6.0, crude protein ≥ 45.0, lysine ≥ 2.8, crude ash ≤ 14.0, crude fiber ≤ 5.0, calcium is about 1.0 - 3.0, total phosphorus ≥ 1.0, sodium chloride is about 0.5 - 3.0, and moisture ≤ 10.0. The Pseudostellaria heterophylla feed additive at 0.5% of the feed quality was added to the feed by spraying, and it was stored in a 4 °C refrigerator for standby.
[0051] 2 Test reagents and instruments
[0052] 2.1 Test reagents
[0053] The main test reagents are shown in Table 1:
[0054] Table 1 Reagents required for the test
[0055]
[0056] 2.2 Test instruments
[0057] The main test instruments are shown in Table 2:
[0058] Table 2-2 Main test instruments
[0059]
[0060] 2.3 Experimental design and aquaculture management
[0061] 2.3.1 Experimental design
[0062] The experiment was carried out in a 1.5T barrel of the recirculating aquaculture system at Ningde Normal University. The blank control group (Group I) was prepared by adding 0.5% of aerated fresh water (by feed quality) to the feed. The Pseudostellaria root extract (concentration: 40mg / mL) prepared in Comparative Example 1 at 0.5% of the feed quality (Group II) was used as the Pseudostellaria control group. The experimental groups were set by adding the Pseudostellaria microecological preparation prepared in Example 1 (Group III), the Pseudostellaria microecological preparation prepared in Example 2 (Group IV), and the Pseudostellaria microecological preparation prepared in Example 3 (Group V) at 0.5% of the feed quality. Each group had 120 tails, and 3 parallel experiments were set. The aquaculture cycle was 60 days.
[0063] 2.3.2 Aquaculture management
[0064] Feeding:
[0065] Feeding was carried out regularly at 8:30 and 17:00 every day. Each time, the bait was 1% - 2% of the fish body weight, and it was adjusted according to the specific feeding situation of the large yellow croaker.
[0066] Aquaculture water environment and daily management:
[0067] The large yellow croaker was cultured using a recirculating aquaculture system. The dissolved oxygen in the aquaculture water body was greater than 5.0mg / L, the pH was controlled between 7.9 and 8.2, the water temperature was maintained between 25.5 and 29°C, and the ammonia nitrogen was kept below 0.2mg / L. The sewage was cleared 2 times a day, the water was changed once, and the water change rate was 50% - 80%. Dead fish were fished out in time, and aquaculture records were made.
[0068] 2.4 Experimental method
[0069] 2.4.1 Biological determination
[0070] After the experiment ended, the feeding of feed was stopped for 1 day. 30 tails were randomly selected from each group to measure (precision: 0.01cm) the body length and weigh (precision: 0.01g) the body weight, visceral weight, and liver weight. The calculation formulas for each parameter are as follows:
[0071] Survival rate (%) = N t / N 0 ×100%;
[0072] Absolute weight gain of body weight (g / d) = (M 2 -M 1 ) / (T 2 -T 1 );
[0073] Absolute growth rate (cm / d) = (L 2 - L 1 ) / (T 2 - T 1 );
[0074] Where: N t is the number of caudal fins at the end; N 0 is the initial number of caudal fins; M is the body weight (g); L is the body length (cm); M 1 , M 2 and L 1 , L 2 are the body weight (g) and body length (cm) at time T 1 , T 2 respectively.
[0075] 2.4.2 Determination of serum enzyme activity indicators
[0076] Collection of serum:
[0077] Stop feeding the feed 1 day before sampling. Randomly select 3 test fish from each parallel group, and collect caudal vein blood samples with a 1 ml sterile syringe. After the collected venous blood is left standing overnight at 4°C, centrifuge it at 4000 r / min and 4°C for 10 min the next day, and take the supernatant and store it frozen in an -80°C refrigerator for the determination of serum enzyme activity indicators.
[0078] Determination of serum enzyme activity:
[0079] The activities of serum superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), acid phosphatase (ACP), alkaline phosphatase (AKP), and lysozyme (LZM) are carried out according to the methods, steps and calculation methods in the kits developed by Nanjing Jiancheng Bioengineering Research Institute.
[0080] 2.4.3 Data statistics and analysis
[0081] The experimental data are expressed as mean ± standard deviation (mean ± SD), and one-way analysis of variance (ANOVA) and Duncan multiple comparison analysis are carried out using SPSS 27.0 software. Column charts are drawn using Origin 2022 software, and p < 0.05 is used as the standard for significant differences.
[0082] 3 Results
[0083] 3.1 Effects on survival rate
[0084] As Figure 1As shown in the figure, after 60 days of cultivation, the survival rate of large yellow croaker in experimental group V was the highest, at (93.33±1.73)%, which was significantly higher than that of other groups (P<0.05); the survival rates of large yellow croaker in experimental groups IV and V were both significantly higher than that of the control group (P<0.05), 11.67% and 16.67% higher than that of control group I respectively, and 2.50% and 7.50% higher than that of control group II; the survival rate of experimental group III was (84.17±2.33)%, 7.50% higher than that of control group I, with significant difference (P<0.05), and 1.67% lower than that of control group II, with no significant difference (P>0.05).
[0085] 3.2 Effects on growth performance
[0086] As Figure 2 shown in Figure A, after 60 days of cultivation, the absolute weight gain rate of control group I was the smallest, at (0.68±0.09) g / d, and that of experimental group V was the largest, at (0.91±0.14) g / d, and it was significantly greater than that of the control group (P<0.05); the absolute weight gain rate of experimental group IV was greater than that of the control group, but the difference was not significant (P>0.05); the absolute weight gain rate of experimental group III was 0.0467 g / d greater than that of control group I and 0.0083 g / d smaller than that of control group II, with no significant difference (P>0.05).
[0087] As Figure 2 shown in Figure B, after 60 days of cultivation, the absolute growth rates of large yellow croaker in the experimental groups were all greater than those in the control group. Among them, the absolute growth rates of experimental groups IV and V were the largest, at (0.13±0.13) cm / d and (0.13±0.02) cm / d respectively, and there was no significant difference between the experimental groups (P>0.05); compared with control group I, the absolute growth rates of experimental groups III, IV, and V increased by 0.027 g / d, 0.036 g / d, and 0.0302 g / d respectively, with significant differences (P<0.05); compared with control group II, the absolute growth rates of experimental groups III, IV, and V increased by 0.0156 g / d, 0.0247 g / d, and 0.01875 g / d respectively. Only the absolute growth rate of experimental group IV and control group II had a significant difference (P<0.05), and there was no significant difference between the other two experimental groups and control group II (P>0.05).
[0088] 3.3 Effects on serum antioxidant enzyme activities
[0089] As Figure 3As shown in
[0090] A, after 60 days of cultivation, the SOD activity in the serum of large yellow croaker in experimental group III was the highest, which was (354.35±1.92) U / mL, but there was no significant difference compared with the control group (P>0.05). It was 14.04 U / mL higher than that in experimental group V, with a significant difference (P<0.05), and 13.24 U / mL higher than that in experimental group IV, with no significant difference (P>0.05). The SOD activity in the serum of experimental group IV was higher than that in control group II and lower than that in control group I, but the difference was not significant (P>0.05). The SOD activity in the serum of experimental group IV was lower than that in the control group, but the difference was not significant (P>0.05).
[0090] As Figure 3 shown in
[0091] B, after 60 days of cultivation, in the CAT activity of large yellow croaker serum, the activities of each experimental group were significantly lower than those of the control group (P<0.05). The activity in experimental group V was the lowest, which was (208.50±13.97) (U / mL). The CAT activity in the serum of control group II was the highest, which was (306.88±16.05) (U / mL) and higher than that in control group I, but the difference was not significant (P>0.05).
[0091] As Figure 3 shown in
[0092] C, after 60 days of cultivation, in the GSH-PX activity of large yellow croaker serum, the activity in experimental group IV was the highest, which was (1148.57.46±91.87) (U / mL); it was significantly higher than that in control group II (P<0.05), and there was no significant difference compared with control group I (P>0.05). The activities in experimental groups III and V were significantly lower than those in control group I (P<0.05). Compared with control group II, the GSH-PX activity in the serum of experimental group III was higher than that in control group II, but the difference was not significant (P>0.05). The activity in experimental group V was lower than that in experimental group II, with a significant difference (P<0.05).
[0092] 4 Effects on serum non-specific immune enzyme activities
[0093] As Figure 4 shown in
[0094] A, after 60 days of cultivation, the ACP activity in the serum of large yellow croaker in experimental group III was (254.27±60.34) (U / L), which was higher than that in the control group. It was 37.40 (U / L) higher than that in control group I, with no significant difference (P>0.05), and 70.75 (U / L) higher than that in control group II, with a significant difference (P<0.05), and was significantly higher than those in experimental groups IV and V (P<0.05). The ACP activities in the sera of experimental groups IV and V were lower than those in the control group. Among them, there was no significant difference between experimental group IV and the control group (P>0.05). The ACP activity in the serum of experimental group V was the lowest, which was (139.55±13.03) (U / L), significantly lower than that in control group I (P<0.05), but there was no significant difference compared with control group II (P>0.05).
[0094] As Figure 4As shown in Figure B, after 60 days of cultivation, the serum AKP activity of large yellow croaker in Control Group II was the highest, at (30.87 ± 6.18) (U / L), while that in Experimental Group V was the lowest, at (26.00 ± 3.82) (U / L); the serum AKP activities of Experimental Groups III and IV were slightly higher than those in Control Group I, and that in Experimental Group V was lower than that in Control Group I, but the differences were not significant (P > 0.05); compared with Control Group II, all experimental groups were lower than Control Group II, but the differences were not significant (P > 0.05), and there were no significant differences in serum AKP activities among all groups (P > 0.05).
[0095] As Figure 4 shown in Figure C, after 60 days of cultivation, the serum LZM activities of large yellow croaker in the experimental groups were all higher than those in the control groups. The serum LZM activity of Experimental Group III was the highest, at (309.49 ± 48.76) U / mL, an increase of 2.07 U / mL compared with Control Group I and an increase of 1.67 U / mL compared with Control Group II, with significant differences (P < 0.05); there was no significant difference between Experimental Group IV and the control groups (P > 0.05); the serum LZM activity of Experimental Group V was significantly higher than that in Control Group I (P < 0.05), and there was no significant difference from Control Group II (P > 0.05).
[0096] It can be seen from Experimental Example 1 that:
[0097] 1. After 60 days of cultivation in this experiment, the Pseudostellariae microecological preparation prepared in Examples 1 - 3 of the present invention has a significant effect on improving the growth performance of large yellow croaker, and the effect is better than that of the Pseudostellariae extract group. This is because after specific types of probiotics are combined with Pseudostellariae in a specific method and specific proportion, a synergistic effect can be achieved, thereby increasing the precipitation of nutrients and active substances such as Pseudostellariae polysaccharides and saponins, facilitating the absorption of large yellow croaker, and promoting growth; it can also produce various digestive enzymes and antibacterial substances, improve the biological activity of digestive enzymes in the digestive tract, enable large yellow croaker to better digest and absorb nutrients in the gastrointestinal tract, improve feed utilization rate, and enhance disease resistance, thus increasing the survival rate of large yellow croaker.
[0098] 2. In this experiment, only the Pseudostellariae microecological preparation fermented by Candida utilis can increase the SOD activity in serum, but the difference is not significant (P > 0.05); the serum CAT enzyme activities of all experimental groups are significantly lower than those in the control group (P < 0.05), indicating that the cultivation time will affect the SOD and CAT activities in the serum of large yellow croaker.
[0099] The serum GSH - Px activity of the Pseudostellariae microecological preparation fermented only by Bacillus subtilis is higher than that in the control group (P < 0.05), not significantly different from the blank control group (P > 0.05), and significantly different from the Pseudostellariae group (P < 0.05). This may be related to the length of cultivation time, experimental animals, types of traditional Chinese medicine, and types of probiotics.
[0100] 3. In this study, adding the microecological preparation of Pseudostellaria heterophylla fermented by Candida utilis to the feed can significantly increase the activities of serum ACP and LZM in large yellow croaker (P < 0.05), and the effect is better after fermentation. However, there is no significant effect on the activity of serum AKP (P > 0.05). This indicates that adding the microecological preparation of Pseudostellaria heterophylla fermented by Candida utilis to the feed according to a specific ratio and method can significantly improve the non-specific immune performance of large yellow croaker, thereby enhancing the fish body's immunity. As can be seen from Experimental Example 1, the LZM activity of large yellow croaker in the Pseudostellaria heterophylla extract group is higher than that in the blank control group, and the microecological preparation of Pseudostellaria heterophylla fermented by Candida utilis can extremely significantly increase the activities of serum ACP and LZM in large yellow croaker (P < 0.01). This may be because probiotics can stimulate the non-specific immune system of the fish body and enhance the body's ability to resist diseases.
[0101] This experiment explored the effects of adding the microecological preparation of Pseudostellaria heterophylla to the feed on the survival rate, growth, and blood immunity of large yellow croaker. The experiment proved that the microecological preparation of Pseudostellaria heterophylla fermented by Lactobacillus plantarum had the best effect on improving the survival rate and growth performance of large yellow croaker. The survival rate was (93.33 ± 1.73)%, the absolute weight gain rate was (0.91 ± 0.14) g / d, and the absolute growth rate was (0.13 ± 0.02) cm / d, all of which were significantly higher than those of the control group (P < 0.05). Through the determination of enzyme activity in the serum, the effects of each example group on the antioxidant enzyme activity in the serum of large yellow croaker were not significant (P > 0.05); the microecological preparation of Pseudostellaria heterophylla fermented by Candida utilis had a promoting effect on the non-specific immunity of large yellow croaker. The serum ACP activity was (254.27 ± 60.34) U / L, which was significantly higher than that of the control group II (P < 0.05), but there was no significant difference compared with the control group I (P > 0.05); the serum AKP activity was (29.02 ± 3.29) U / L, and there was no significant difference compared with the control group (P > 0.05); the serum LZM activity was (309.49 ± 48.76) U / mL, which was significantly higher than that of the control group (P < 0.05).
[0102] The embodiments of the present application have been described above in conjunction with the accompanying drawings. However, the present application is not limited to the above specific embodiments. The above specific embodiments are merely illustrative and not restrictive. Under the inspiration of the present application, those of ordinary skill in the art can also make many forms without departing from the purpose of the present application and the scope protected by the claims, and all of them belong to the protection scope of the present application.
Claims
1. A method for preparing a probiotic preparation of Radix Pseudostellariae Radix, characterized in that: The following steps are involved: (1) Preparation of Pseudostellariae Radix Root Extract: Wash and air-dry the root tubers of Pseudostellaria heterophylla, grind them into powder, pass through a 50-70 mesh sieve, and place the powder in a round-bottom flask, add 75-85% ethanol, reflux extract in a water bath at 90±1℃ for 2-4h, filter while hot, extract the residue with 95±1℃ ultrapure water for 3-5h and filter, mix and concentrate the two filtrates, remove water, and obtain the root extract of Pseudostellaria heterophylla, which is stored in a refrigerator at 4±1℃ for later use; (2) The Pseudostellariae Radix Root Extract was fermented in a probiotic shaker for 23-25 hours to prepare the Pseudostellariae Radix Probiotics Preparation, which was then placed in a refrigerator at 4±1℃ for later use.
2. The method for preparing the Pseudostellariae Radix microecological preparation according to claim 1, wherein: The concentration of the Pseudostellaria heterophylla root extract is 35-45 mg / mL.
3. The method for preparing the Pseudostellariae Radix microecological preparation according to claim 1, wherein: The inoculation amount of probiotics is 4-6%.
4. The method for preparing the Pseudostellariae Radix microecological preparation according to claim 1, wherein: The probiotics are Candida utilis, Bacillus subtilis or plant lactic acid bacteria.
5. The method for preparing the Pseudostellariae Radix microecological preparation according to claim 4, characterized in that: The Candida utilis is cultured at 30±1° C. and 190-210 r / min.
6. The method for preparing the Pseudostellariae Radix microecological preparation according to claim 4, characterized in that: The bacillus subtilis and plant lactic acid bacteria are cultured at 37±1°C and 170-190r / min.
7. A Pseudostellariae Radix microecological preparation, characterized in that: The preparation method is described in any one of claims 1 to 5.
8. Application of the Pseudostellaria heterophylla microecological preparation according to claim 7 as a feed additive for large yellow croaker.
9. A large yellow croaker feed, characterized in that: Including the Pseudostellariae Radix microecological preparation described in claim 7.
10. The large yellow croaker feed according to claim 9, characterized in that: The added amount of the Pseudostellaria heterophylla microecological preparation is 0.4-0.6% of the feed mass.