CN114947012B - Rana nigromaculata feed additive and preparation method and application thereof - Google Patents
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Abstract
The application provides a rana nigromaculata feed additive, and a preparation method and application thereof. The application relates to a rana nigromaculata feed additive, which comprises radix codonopsis pilosulae extract, eucommia ulmoides extract and zeolite powder; the black spot frog feed additive disclosed by the application can be added into feed, so that the cataract infection mortality rate of the cultured black spot frog can be obviously reduced, the liver state and intestinal mucosa morphology can be improved, and the relative abundance of beneficial bacteria Lactococcus lactis can be obviously increased.
Description
Technical Field
The application relates to the technical field of feed additives, in particular to a rana nigromaculata feed additive and a preparation method and application thereof.
Background
The black frog (Rana nigromaculata) is an amphibian of Rana, belonging to the family Ranae and genus Rana, commonly called as "farmland guard", "crop protection god", and has strong nature and extremely strong jumping ability, and is an energy hand for catching more than 30 rice pests such as borer moth, rice bud, locust, mole cricket, leafhopper, cricket and the like, more than 70 daily pests are killed by the daily pests, and the pest is killed by the daily life. Because the rana nigromaculata has delicious taste, rich nutrition and medical value, the market demand of the rana nigromaculata is larger.
In recent years, the rapid development of the rana nigromaculata feed industry is a decisive driving force for the rapid development of the rana nigromaculata breeding industry. Although the artificial high-density culture greatly improves the productivity, the rana nigromaculata is affected by density stress, the physical function is poor, and the high nutrition of the artificial feed leads to overnutrition in the rana nigromaculata, high metabolic pressure and symptoms such as fatty liver and the like which commonly exist, causes frequent occurrence of various diseases and has high transmission speed, and especially, the head-tilting disease can cause a large amount of death and huge economic loss. Under the conditions of feed resistance and disease stress, the phenomenon that the cultured rana nigromaculata has multiple diseases is caused, and a breeder takes the medicines such as antibiotics into consideration for prevention and treatment at first, so that the dosage of the antibiotics for treatment at the breeding end is increased, thereby indirectly threatening the health of human beings. Therefore, the development of natural, nontoxic, high-efficiency and low-cost green feed additives is an urgent need of the rana nigromaculata breeding industry. The plant extract is used as a novel green feed additive, is applied to livestock, poultry, fish and shrimp to be mature, and is also a trend in the field of the rana nigromaculata additive.
However, the existing technology about the rana nigromaculata plant/Chinese herbal medicine extract additive is very rare, and the solution for preventing cataract and fatty liver in the feeding process of rana nigromaculata by using plant extracts is not yet available.
Disclosure of Invention
In view of the above, the present application provides a rana nigromaculata feed additive, and a preparation method and application thereof, so as to solve the above problems or at least partially solve the above problems.
In a first aspect, the application provides a rana nigromaculata feed additive, which comprises the following raw materials, by mass, 30-55% of codonopsis pilosula extract, 25-55% of eucommia ulmoides extract and 10-40% of zeolite powder.
Preferably, the rana nigromaculata feed additive comprises the following raw materials in percentage by mass: 35-55% of radix codonopsis extract, 35-55% of eucommia ulmoides extract and 10-20% of zeolite powder.
Preferably, the rana nigromaculata feed additive comprises the following raw materials, by mass, 45% of codonopsis pilosula extract, 45% of eucommia ulmoides extract and 10% of zeolite powder.
Preferably, the zeolite powder of the rana nigromaculata feed additive has a mesh number of 60-120 meshes.
In a second aspect, the application also provides a preparation method of the rana nigromaculata feed additive, which comprises the following steps:
grinding radix Codonopsis, adding into water, heating, filtering to remove residue, concentrating the filtrate under heating, adding ethanol, filtering, removing residue, distilling under reduced pressure, and drying to obtain radix Codonopsis extract;
grinding folium Eucommiae, adding into water, heating, filtering to remove residue, concentrating the filtrate under heating, adding ethanol, filtering, removing residue, distilling under reduced pressure, and drying to obtain cortex Eucommiae extract;
mixing radix Codonopsis extract, eucommiae cortex extract and zeolite powder at a certain proportion to obtain the final product.
Preferably, the preparation method of the rana nigromaculata feed additive comprises the steps of grinding 0.5-2 kg of radix codonopsis pilosulae, adding into 5-15L of water, heating at 90-100 ℃ for 1-3 h, filtering to remove filter residues, heating and concentrating filtrate to 0.5-1.5L, adding absolute ethyl alcohol, filtering, removing filter residues, distilling under reduced pressure, and drying to obtain radix codonopsis pilosulae extract.
Preferably, the preparation method of the rana nigromaculata feed additive comprises the steps of grinding 0.5-2 kg of eucommia ulmoides leaves, adding into 5-15L of water, heating at 90-100 ℃ for 1-3 h, filtering to remove filter residues, heating and concentrating filtrate to 0.5-1.5L, adding absolute ethyl alcohol, filtering, removing filter residues, distilling under reduced pressure, and drying to obtain eucommia ulmoides extract.
Preferably, the preparation method of the rana nigromaculata feed additive comprises the following steps of: sieving the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder with a 60-80 mesh sieve respectively, and uniformly mixing the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder which are sieved according to a proportion.
In a third aspect, the application also provides an application of the rana nigromaculata feed additive or the rana nigromaculata feed additive prepared by the preparation method in preparing medicines for treating cataract and fatty liver of rana nigromaculata.
Compared with the prior art, the rana nigromaculata feed additive has the following beneficial effects:
1. the application relates to a rana nigromaculata feed additive, which comprises radix codonopsis pilosulae extract, eucommia ulmoides extract and zeolite powder; the black spot frog feed additive disclosed by the application can be added into feed, so that the cataract infection mortality rate of the cultured black spot frog can be obviously reduced, the liver state and intestinal mucosa morphology can be improved, and the relative abundance of potential beneficial bacteria Lactococcus lactis can be obviously increased.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It is evident that the drawings in the following description are only some embodiments of the present application and that other drawings may be obtained from these drawings without inventive effort for a person of ordinary skill in the art.
FIG. 1 mortality of the rana nigromaculata cataracts in the control group, the test 1 group and the test 2 group after the cultivation test is completed;
FIG. 2 morphology of the normal liver, the liver of the flower and the liver of the yellow of the rana nigromaculata;
FIG. 3 is the ratio of normal liver, liver of flower and liver of Rana nigromaculata in the control group, the test 1 group and the test 2 group after the cultivation test is completed;
FIGS. 4 to 6 are graphs showing the intestinal H.E staining of the control group, the test 1 group and the test 2 group of Rana nigromaculata after the cultivation test is completed;
FIG. 7 shows the intestinal microbial structure changes of the control group, the test 1 group and the test 2 group of rana nigromaculata after the cultivation test.
Detailed Description
The following description of the embodiments of the present application will be made in detail and with reference to the embodiments of the present application, but it should be apparent that the described embodiments are only some embodiments of the present application, and not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present application without making any inventive effort, are intended to fall within the scope of the present application.
The embodiment of the application provides a rana nigromaculata feed additive, which comprises the following raw materials, by mass, 30-55% of codonopsis pilosula extract, 25-55% of eucommia ulmoides extract and 10-40% of zeolite powder.
Specifically, radix Codonopsis extract has cataract pathogen antagonism effect, cortex Eucommiae extract has liver and gallbladder protecting effect, and zeolite powder is used as carrier.
In some embodiments, the method comprises the following raw materials in percentage by mass: 35-55% of radix codonopsis extract, 35-55% of eucommia ulmoides extract and 10-20% of zeolite powder.
In some embodiments, the composition comprises the following raw materials, by mass, 45% of radix codonopsis extract, 45% of eucommia ulmoides extract and 10% of zeolite powder.
In some embodiments, the zeolite powder has a mesh size of 60 to 120 mesh.
The rana nigromaculata feed additive comprises radix codonopsis pilosulae extract, eucommia ulmoides extract and zeolite powder, and can obviously reduce the cataract infection mortality rate of cultured rana nigromaculata, improve the liver state and intestinal mucosa morphology and obviously increase the relative abundance of beneficial bacteria Lactococcus lactis when being added into feed.
Based on the same inventive concept, the embodiment of the application also provides a preparation method of the rana nigromaculata feed additive, which comprises the following steps:
s1, grinding codonopsis pilosula, adding the ground codonopsis pilosula into water, heating, filtering to remove filter residues, heating and concentrating filtrate, adding ethanol, filtering to remove filter residues, distilling under reduced pressure, and drying to obtain the codonopsis pilosula extract;
s2, grinding eucommia ulmoides leaves, adding the ground eucommia ulmoides leaves into water, heating, filtering to remove filter residues, heating and concentrating filtrate, adding ethanol, filtering to remove filter residues, distilling under reduced pressure, and drying to obtain an eucommia ulmoides extract;
s3, uniformly mixing the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder according to a proportion to obtain the rana nigromaculata feed additive.
In some embodiments, 0.5-2 kg of radix codonopsis pilosulae is ground, added into 5-15L of water, heated at 90-100 ℃ for 1-3 h, filtered to remove filter residues, heated and concentrated to 0.5-1.5L, added with absolute ethyl alcohol to make the mass content of alcohol reach 70%, filtered to remove filter residues, distilled at room temperature under reduced pressure until the radix codonopsis pilosulae extract is completely dried, and the radix codonopsis pilosulae extract is obtained.
In some embodiments, 0.5-2 kg of eucommia leaves are ground, added into 5-15L of water, heated for 1-3 h at 90-100 ℃, filtered to remove filter residues, heated and concentrated to 0.5-1.5L, added with absolute ethyl alcohol to enable the mass content of alcohol to reach 70%, filtered, removed filter residues, distilled at room temperature under reduced pressure until the eucommia extracts are completely dried, and finally the eucommia extracts are obtained.
In some embodiments, before mixing the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder uniformly according to the proportion, the method further comprises: sieving the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder with a 60-80 mesh sieve respectively, and uniformly mixing the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder which are sieved according to a proportion.
Specifically, the codonopsis pilosula extract, the eucommia ulmoides extract and the zeolite powder which are sieved are added into a dry powder stirrer according to the proportion and stirred for 20-30 min, so that the codonopsis pilosula extract, the eucommia ulmoides extract and the zeolite powder are fully and uniformly mixed, and the rana nigromaculata feed additive is obtained.
Based on the same inventive concept, the embodiment of the application also provides the application of the rana nigromaculata feed additive or the rana nigromaculata feed additive prepared by the preparation method in preparing medicaments for treating cataract and fatty liver of rana nigromaculata.
The frog feed additive of the present application is further described in the following specific examples. This section further illustrates the summary of the application in connection with specific embodiments, but should not be construed as limiting the application. The technical means employed in the examples are conventional means well known to those skilled in the art, unless specifically stated. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present application are those conventional in the art.
Example 1
The embodiment provides a rana nigromaculata feed additive, which comprises the following raw materials by mass percent, 45% of radix codonopsis pilosulae extract, 45% of eucommia ulmoides extract and 10% of zeolite powder.
The preparation method of the rana nigromaculata feed additive comprises the following steps:
s1, grinding 1kg of radix codonopsis pilosulae, adding into 10L of water, heating at 95 ℃ for 2 hours, filtering to remove filter residues, heating and concentrating filtrate to 1L, adding absolute ethanol to enable the mass content of alcohol to reach 70%, filtering to remove the filter residues, and distilling at room temperature under reduced pressure until the filter residues are completely dried to obtain radix codonopsis pilosulae extract;
s2, grinding 1kg of eucommia ulmoides leaves, adding into 10L of water, heating at 95 ℃ for 2 hours, filtering to remove filter residues, heating and concentrating filtrate to 1L, adding absolute ethanol to enable the mass content of alcohol to reach 70%, filtering, removing the filter residues, and distilling at room temperature under reduced pressure until the eucommia ulmoides extracts are completely dried to obtain the eucommia ulmoides extracts;
s3, screening the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder by a 60-mesh sieve respectively, and adding the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder which are screened by the sieve into a dry powder stirrer according to the mass ratio, stirring for 30min, and fully and uniformly mixing to obtain the rana nigromaculata feed additive.
Example 2
The embodiment provides a rana nigromaculata feed additive, which comprises the following raw materials in percentage by mass of 55% of codonopsis pilosula extract, 35% of eucommia ulmoides extract and 10% of zeolite powder.
The preparation method of the rana nigromaculata feed additive is the same as that of example 1.
Example 3
The embodiment provides a rana nigromaculata feed additive, which comprises the following raw materials by mass percent, 40% of radix codonopsis pilosulae extract, 40% of eucommia ulmoides extract and 20% of zeolite powder.
The preparation method of the rana nigromaculata feed additive is the same as that of example 1.
Cultivation test
The average body mass of the rana nigromaculata is 20.35+/-1.27 g in the culture test and effect test, and the test site is located at the rice frog culture base of the family and professionals in Yongxin county high-city county in Jiangxi province. Satiety feeding is performed at 6:30 a.m. and 17:30 a.m. each day. The test rana nigromaculata was randomly divided into three groups, control group, test 1 group and test 2 group, and the number of rana nigromaculata in each group was 1400. The control group was fed with commercial feed, and the test 1 group and the test 2 group were fed with the rana nigromaculata feed additive prepared in example 1 with an additive mass fraction of 1.25% and 2.5% (relative to commercial feed), respectively.
After the cultivation test is finished, the changes of cataract mortality, liver morphology, intestinal mucosa morphology and microbial structure of each group of rana nigromaculata are counted and analyzed.
Figure 1 shows the mortality rate (mortality rate) of rana nigromaculata cataracts in the control group, the test 1 group (i.e., test 1 in figure 1) and the test 2 group (i.e., test 2 in figure 1) after the end of the culture test.
As can be seen from FIG. 1, in example 1 of the commercial feed addition for Rana nigromaculata, the lethality of cataract of Rana nigromaculata was reduced from 17.14% (control group) to 10.09% (test group 1) and 8.26% (test group 2).
FIG. 2 morphology of the normal liver, the liver of the Bullfrog, and the liver of the Bullfrog.
FIG. 3 shows the proportions of normal liver, liver of flower and liver of Rana nigromaculata in the control group, the test 1 group and the test 2 group after the cultivation test.
As can be seen from fig. 3, the normal proportion of the liver of rana nigromaculata in the test group 2 was significantly increased and the proportion of the liver of floriated and yellow liver was significantly decreased.
FIGS. 4 to 6 are graphs showing the intestinal H.E staining of the control, test 1 and test 2 group of Rana nigromaculata after the cultivation test.
From FIGS. 4 to 6, it can be seen that the intestinal microvilli structure and intestinal epithelial cell lamina propria of the group 2 Rana nigromaculata were more complete.
FIG. 7 and Table 1 show the changes in intestinal microbial structures of the control group, the test 1 group and the test 2 group of Rana nigromaculata after the cultivation test is completed.
As can be seen from fig. 7 and table 1, the Lactococcus lactis relative abundance in test group 2 was significantly higher than that in the control group. Lactococcus is a common probiotic for aquatic animals and has the functions of promoting immunity and improving disease resistance for hosts.
TABLE 1 different treatments for the horizontal structure of intestinal microorganisms of Rana nigromaculata
| Belonging to the genus | Control | Test 1 | Test 2 |
| Cetobacterium | 29.33±33.73 | 4.90±4.82 | 10.83±6.43 |
| Bacteroides | 13.98±11.10 a | 5.50±5.87 ab | 4.30±2.85 b |
| Romboutsia | 6.05±6.01 | 2.83±3.69 | 3.90±2.92 |
| Weissella | 0.58±0.53 | 12.65±24.50 | 4.15±4.65 |
| Prevotella_9 | 0.24±0.22 | 0.06±0.03 | 4.50±6.33 |
| Lactococcus | 1.20±0.92 a | 1.97±2.95 a | 9.61±8.42 b |
| Bilophila | 0.95±0.63 | 10.53±11.81 | 0.89±1.10 |
| Akkermansia | 8.24±10.94 | 2.23±1.80 | 0.73±0.51 |
| Clostridium_sensu_stricto_1 | 2.54±4.32 | 1.37±1.68 | 6.95±9.77 |
| Desulfovibrio | 2.99±3.84 | 4.13±3.97 | 0.74±0.52 |
In Table 1, the same letters or no letters in the same row of data indicate that the difference is not significant (P > 0.05), and the different letters in the same row indicate that the difference is significant (P < 0.05).
The test results show that the addition of 1.25-2.5% of the brown spot frog feed additive in the embodiment 1 into the brown spot frog commercial feed can obviously reduce the cataract infection mortality rate of cultured brown spot frog, improve the liver state and intestinal mucosa morphology and obviously increase the relative abundance of potential beneficial bacteria Lactococcus lactis. Therefore, the application has important application value for the healthy development of the rana nigromaculata breeding industry.
The foregoing description of the preferred embodiments of the application is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the application.
Claims (4)
1. Application of the composition in preparing medicines for treating cataract and fatty liver of Rana nigromaculata;
wherein the composition comprises the following raw materials in percentage by mass: 45% of radix codonopsis extract, 45% of eucommia ulmoides extract and 10% of zeolite powder;
the preparation method of the radix codonopsis extract comprises the following steps:
grinding 0.5-2 kg of radix codonopsis pilosulae, adding into 5-15L of water, heating at 90-100 ℃ for 1-3 h, filtering to remove filter residues, heating and concentrating filtrate to 0.5-1.5L, adding absolute ethyl alcohol, filtering, removing filter residues, distilling under reduced pressure, and drying to obtain radix codonopsis pilosulae extract;
the preparation method of the eucommia ulmoides extract comprises the following steps:
grinding 0.5-2 kg of eucommia leaves, adding into 5-15L of water, heating at 90-100 ℃ for 1-3 h, filtering to remove filter residues, heating and concentrating the filtrate to 0.5-1.5L, adding absolute ethyl alcohol, filtering, removing filter residues, distilling under reduced pressure, and drying to obtain the eucommia extract.
2. The use according to claim 1, wherein the zeolite powder has a mesh size of 60 to 120 mesh.
3. The use according to claim 1, wherein the method of preparing the composition comprises the steps of:
mixing radix Codonopsis extract, eucommiae cortex extract and zeolite powder at a certain proportion to obtain the final product.
4. The use according to claim 3, wherein before mixing the radix codonopsis extract, the eucommia ulmoides extract and the zeolite powder uniformly according to the mixture ratio, the method further comprises: sieving radix Codonopsis extract, eucommiae cortex extract and zeolite powder with 60-80 mesh sieve, and mixing the sieved radix Codonopsis extract, eucommiae cortex extract and zeolite powder at a certain proportion.
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| CN108813127A (en) * | 2018-07-28 | 2018-11-16 | 厦门同欣荣饲料科技有限公司 | A kind of frog feed and preparation method thereof |
| CN109221602A (en) * | 2018-07-10 | 2019-01-18 | 枞阳县鑫誉黑斑蛙养殖有限责任公司 | A kind of Rana nigromaculata special feed |
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| CN106616068A (en) * | 2017-02-28 | 2017-05-10 | 唐人神集团股份有限公司 | Feed additive for rana nigromaculata and preparation method thereof |
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| CN108576407A (en) * | 2018-04-18 | 2018-09-28 | 林世琛 | A kind of frog expanded pellet feed |
| CN109221602A (en) * | 2018-07-10 | 2019-01-18 | 枞阳县鑫誉黑斑蛙养殖有限责任公司 | A kind of Rana nigromaculata special feed |
| JP2020010673A (en) * | 2018-07-17 | 2020-01-23 | 仁助 中山 | Circulation system of breeding water for culturing edible frog, circulation system of breeding water for culturing feed of edible frog, feeding method of feed for edible frog and production method of feed |
| CN108813127A (en) * | 2018-07-28 | 2018-11-16 | 厦门同欣荣饲料科技有限公司 | A kind of frog feed and preparation method thereof |
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