CN113712134A

CN113712134A - Chinese herbal medicine immunopotentiator for groupers and preparation method and application thereof

Info

Publication number: CN113712134A
Application number: CN202111036373.0A
Authority: CN
Inventors: 汤菊芬; 简纪常; 蔡佳; 黄瑜; 王蓓; 黄郁葱; 蔡双虎; 鲁义善
Original assignee: Guangdong Ocean University
Current assignee: Guangdong Ocean University
Priority date: 2021-09-06
Filing date: 2021-09-06
Publication date: 2021-11-30

Abstract

The invention discloses a Chinese herbal medicine immunopotentiator for groupers as well as a preparation method and application thereof, belonging to the technical field of aquatic product culture. The Chinese herbal medicine immunopotentiator for the grouper comprises the following raw materials in parts by weight: 5-15 parts of prepared rehmannia root, 10-30 parts of mung bean, 5-20 parts of kudzu root, 1-10 parts of poria cocos, 1-10 parts of liquorice, 1-10 parts of radix curcumae, 1-10 parts of bighead atractylodes rhizome, 5-15 parts of rhizoma acori graminei, 1-10 parts of albizia flower, 1-10 parts of fructus forsythiae, 5-20 parts of dandelion, 5-10 parts of longan seed, 1-10 parts of costus root and 1-15 parts of hawthorn. The prepared Chinese herbal medicine immunopotentiator has obvious growth promoting and disease resisting effects on the grouper, is convenient to use and can be used as a feed additive.

Description

Chinese herbal medicine immunopotentiator for groupers and preparation method and application thereof

Technical Field

The invention relates to the technical field of aquatic product culture, in particular to a Chinese herbal medicine immunopotentiator for groupers and a preparation method and application thereof.

Background

The grouper is an important economic fish for seawater cage culture in south China, has the characteristics of rapid growth, strong disease resistance, delicious meat quality, high nutritional value, suitability for large-scale culture and the like, and is deeply popular in the market. In recent years, as the breeding efficiency is excessively pursued, the breeding environment is gradually deteriorated, and the disease problem frequently occurs. Vibrio harveyi (Vibrio harveyi) is the main pathogen of vibriosis of rockfishes, often causes deep skin ulcer, gastrointestinal diseases and eye diseases of diseased fish, leads to the increase of morbidity and mortality of rockfishes, and causes great economic loss.

At present, antibiotics are mainly applied to the culture of groupers to treat vibriosis, but abuse of the antibiotics causes ecological problems such as appearance of drug-resistant bacteria, drug residue and the like, and the edible safety of the groupers is influenced. Although the vaccine has a specific effect on preventing the vibriosis of the grouper, a series of technical bottlenecks exist for large-scale popularization and application, such as: the injection method is time-consuming and labor-consuming in operation and difficult to popularize and apply (especially seawater cage culture); the oral vaccine has the problems of immature production technology, high production cost and the like.

The Chinese herbal medicine is a green natural medicine, has the advantages of small toxic and side effects, multiple components, multiple targets and multiple effects, difficulty in inducing pathogenic bacteria drug resistance, low pollution, low price, rich resources and the like, and can be used in the fields of disease control of aquaculture and the like. At present, Chinese herbal medicines for preventing and treating aquatic diseases are used as immunopotentiators or anti-pathogenic medicines, and during feeding, the problems of high average feed coefficient, low survival rate and unsatisfactory immune protection effect exist. For example, patent CN106540157A provides a compound Chinese herbal medicine immunopotentiator for grouper, which comprises the following raw materials: 10% of chrysanthemum powder, 8% of perilla frutescens powder, 10% of seaweed powder, 8% of mulberry leaf powder, 10% of carrot powder, 8% of moringa oleifera powder, 5% of hairyvein agrimony, 7% of purple perilla, 5% of astragalus membranaceus, 5% of glabrous greenbrier rhizome, 3% of houttuynia cordata, 5% of radix stemonae, 3% of honeysuckle, 5% of bighead atractylodes rhizome, 3% of divaricate saposhnikovia root, 3% of garlicin, 0.5% of low chitosan and 1.5% of dendrobium officinale. The culture result of the grouper shows that the bait coefficient is higher, the activity of superoxide dismutase in the grouper serum is lower, and the disease resistance of the grouper is to be further improved. Therefore, the development of the plant Chinese herbal medicine immunopotentiator which is green, safe and excellent in immune effect has very important significance.

Disclosure of Invention

The Chinese herbal medicine immunopotentiator has obvious effects of promoting growth and resisting diseases on the grouper, is convenient to use, can be used as a feed additive, and overcomes the defects and shortcomings of the existing method for preventing and treating vibriosis of the mariculture grouper.

In order to achieve the purpose, the invention provides the following scheme:

the invention provides a Chinese herbal medicine immunopotentiator for grouper, which comprises the following raw materials in parts by weight: 5-15 parts of prepared rehmannia root, 10-30 parts of mung bean, 5-20 parts of kudzu root, 1-10 parts of poria cocos, 1-10 parts of liquorice, 1-10 parts of radix curcumae, 1-10 parts of bighead atractylodes rhizome, 5-15 parts of rhizoma acori graminei, 1-10 parts of albizia flower, 1-10 parts of fructus forsythiae, 5-20 parts of dandelion, 5-10 parts of longan seed, 1-10 parts of costus root and 1-15 parts of hawthorn.

Further, the feed comprises the following raw materials in parts by weight: 8-10 parts of prepared rehmannia root, 15-20 parts of mung bean, 10-15 parts of kudzu root, 5-8 parts of poria cocos, 5-7 parts of liquorice, 4-6 parts of radix curcumae, 4-7 parts of bighead atractylodes rhizome, 6-10 parts of rhizoma acori graminei, 5-8 parts of albizia flower, 4-6 parts of fructus forsythiae, 8-12 parts of dandelion, 7-9 parts of longan seed, 7-9 parts of costus root and 8-12 parts of hawthorn.

The invention provides a preparation method of a Chinese herbal medicine immunopotentiator for grouper, which comprises the following steps: 14 Chinese herbal medicines of prepared rehmannia root, mung bean, kudzu root, poria with poria cocos, liquorice, radix curcumae, bighead atractylodes rhizome, rhizoma acori graminei, albizia flower, fructus forsythiae, dandelion, longan seed, elecampane and hawthorn are respectively crushed and uniformly mixed in proportion to obtain the Chinese herbal medicine immunopotentiator for grouper.

Further, the particle size of the crushed material is required to be 100-mesh sieved.

The invention also provides application of the Chinese herbal medicine immunopotentiator for the groupers as a feed additive.

Further, the Chinese herbal medicine immunopotentiator is added into the grouper feed, and accounts for 0.5-2% of the mass of the grouper feed.

The invention discloses the following technical effects:

the invention provides a Chinese herbal medicine immunopotentiator for grouper, a preparation method and application thereof.

The Chinese herbal medicine immunopotentiator is added into the grouper compound feed by 0.5-2 percent, and the result shows that the grouper compound feed is fed for 28 days: 1.5 percent of Chinese herbal medicine immunopotentiator is added into the basic feed, so that the weight gain rate and the survival rate of the groupers in a test group are highest, and the feed coefficient is lowest; the immune indexes such as leukocyte phagocytosis rate, superoxide dismutase activity, lysozyme activity, antibacterial activity and the like in the blood of the grouper are obviously higher than those of a control group, the number of probiotics such as bifidobacteria, lactic acid bacteria and the like in intestinal tracts is obviously higher than that of the control group, and the number of pathogenic bacteria such as escherichia coli, vibrio and the like is obviously lower than that of the control group. After the grouper is infected with vibrio harveyi by artificial injection, the immune protection rate is 74.16%. The Chinese herbal medicine immunopotentiator provided by the invention can obviously improve the intestinal flora structure of the grouper, improve the immune function and vibrio infection resistance of the grouper and improve the culture survival rate.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 shows the effect of Chinese herbal immunopotentiators on the total number of intestinal bacteria in Epinephelus;

FIG. 2 shows the effect of Chinese herbal immunopotentiators on the number of Vibrio enterobacter of Epinephelus;

FIG. 3 shows the effect of the Chinese herbal immunopotentiator on the number of Escherichia coli in intestinal tract of Epinephelus;

FIG. 4 shows the effect of the Chinese herbal immunopotentiator on the number of lactic acid bacteria in the intestinal tract of Epinephelus;

FIG. 5 shows the effect of the Chinese herbal immunopotentiator on the amount of Bifidobacterium in intestinal tract of Epinephelus;

FIG. 6 shows the effect of the Chinese herbal immunopotentiator on the phagocytosis rate of blood leukocytes of grouper;

FIG. 7 shows the effect of the Chinese herbal immunopotentiator on the serum lysozyme activity of Epinephelus;

FIG. 8 shows the effect of the Chinese herbal immunopotentiator on the activity of serum SOD of Epinephelus;

FIG. 9 shows the effect of the Chinese herbal immunopotentiator on the antibacterial activity of the serum of Epinephelus;

FIG. 10 shows the effect of Chinese herbal immunopotentiators on the immunoprotection rate of Epinephelus.

Detailed Description

Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The description and examples are intended to be illustrative only.

As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.

Example 1

Taking 10 parts of prepared rehmannia root, 20 parts of mung bean, 10 parts of kudzuvine root, 5 parts of poria cocos, 5 parts of liquorice, 5 parts of turmeric root-tuber, 5 parts of bighead atractylodes rhizome, 10 parts of grassleaf sweelflag rhizome, 5 parts of albizia flower, 5 parts of weeping forsythia, 10 parts of dandelion, 8 parts of longan seed, 5 parts of costustoot and 10 parts of hawthorn fruit, respectively crushing, sieving with a 100-mesh sieve, and uniformly mixing to obtain the Chinese herbal medicine immunopotentiator for the grouper.

Example 2

5 parts of prepared rehmannia root, 30 parts of mung bean, 5 parts of kudzuvine root, 10 parts of poria cocos, 1 part of liquorice, 10 parts of turmeric root-tuber, 1 part of bighead atractylodes rhizome, 15 parts of grassleaf sweelflag rhizome, 1 part of albizia flower, 10 parts of weeping forsythia capsule, 5 parts of dandelion, 10 parts of longan seed, 1 part of costustoot and 15 parts of hawthorn are respectively crushed and sieved by a 100-mesh sieve and mixed uniformly to obtain the Chinese herbal medicine immunopotentiator for grouper.

Example 3

Taking 15 parts of prepared rehmannia root, 10 parts of mung bean, 20 parts of kudzuvine root, 1 part of poria cocos, 10 parts of liquorice, 1 part of radix curcumae, 10 parts of bighead atractylodes rhizome, 5 parts of rhizoma acori graminei, 10 parts of albizia flower, 1 part of fructus forsythiae, 20 parts of dandelion, 5 parts of longan seed, 10 parts of elecampane and 1 part of hawthorn, respectively crushing, sieving with a 100-mesh sieve, and uniformly mixing to obtain the Chinese herbal medicine immunopotentiator for the grouper.

Example 4

8 parts of prepared rehmannia root, 20 parts of mung bean, 15 parts of kudzuvine root, 5 parts of poria cocos, 7 parts of liquorice, 4 parts of turmeric root-tuber, 7 parts of bighead atractylodes rhizome, 6 parts of grassleaf sweelflag rhizome, 8 parts of albizia flower, 4 parts of weeping forsythia capsule, 12 parts of dandelion, 7 parts of longan seed, 9 parts of costustoot and 8 parts of hawthorn are respectively crushed and sieved by a 100-mesh sieve, and the crushed materials are uniformly mixed to obtain the Chinese herbal medicine immunopotentiator for the grouper.

Example 5

Taking 10 parts of prepared rehmannia root, 15 parts of mung bean, 10 parts of kudzuvine root, 8 parts of poria cocos, 5 parts of liquorice, 6 parts of turmeric root-tuber, 4 parts of bighead atractylodes rhizome, 10 parts of grassleaf sweelflag rhizome, 5 parts of albizia flower, 6 parts of weeping forsythia capsule, 8 parts of dandelion, 9 parts of longan seed, 7 parts of costustoot and 12 parts of hawthorn fruit, respectively crushing, sieving with a 100-mesh sieve, and uniformly mixing to obtain the Chinese herbal medicine immunopotentiator for grouper.

Comparative example 1

The difference from example 1 is that rehmanniae radix Preparata was not added.

Comparative example 2

The difference from example 1 is that kudzu root and curcuma aromatica were not added.

Comparative example 3

The difference from embodiment 1 is that a Poria cocos is not added.

Comparative example 4

The difference from example 1 is that no Acorus gramineus soland is added.

Comparative example 5

The difference from example 1 is that licorice and forsythia suspensa were not added.

Comparative example 6

The difference from example 1 is that dandelion was not added.

Comparative example 7

The difference from example 1 is that longan seeds were not added.

Comparative example 8

The difference from example 1 is that no radix aucklandiae was added.

Comparative example 9

The same as example 1, except that the licorice was replaced with honeysuckle flower.

Comparative example 10

The same as example 1, except that the silk tree flower was replaced with chrysanthemum powder.

Test example 1

The Chinese herbal medicine immunopotentiator for grouper in example 1 was added to grouper feed (commercially available conventional grouper feed) in a ratio of 0.5%, 1.0%, 1.5% and 2.0% (mass percentage) to be mixed uniformly to prepare 4 kinds of granular medicinal baits.

The prepared medicinal bait is used for feeding the margarita, gentian and grouper (the grouper is cultured in a high-tech garden of Guangdong ocean university in east island of Zhanjiang city, three times are respectively arranged in a test group and a control group), the control group is fed with commercial feed, the test group is respectively fed with 4 kinds of medicinal bait feed (respectively marked as 0.5 group, 1.0 group, 1.5 group and 2.0 group), and the feeding management is carried out according to a scientific feeding method. The oxygen machine supplies oxygen, and water is changed for 1 time every two days. The daily feeding amount is 3-5% of the fish weight, and the feeding is carried out twice (9 am, 17 pm) in a feeding period of 28 days.

On the 28 th day, the fish of the test group and the fish of the control group are respectively weighed and counted, 15 tail groupers are randomly taken from each group, the tail vein is sampled, and then the intestinal tract is taken out under the aseptic condition. Respectively measuring the phagocytosis rate of leucocytes, the activity of superoxide dismutase, the content of lysozyme, the antibacterial activity and other immune indexes in blood and the number of intestinal flora.

After 29 days, use 1X 10⁷cells/mL of Vibrio harveyi bacterial suspension is subjected to toxicity attack, the injection dosage is 0.1 mL/tail, the death condition of each group of groupers within 10 days is observed and recorded, and the immune protection rate is calculated.

The results show that:

1) effect of Chinese herbal medicine immunopotentiator on growth of grouper

The influence of the Chinese herbal medicine immunopotentiator on the growth of the groupers is shown in table 1, the weight gain rate, the survival rate and the feed utilization rate of 4 test groups of groupers are obviously higher than those of a control group, and the growth promoting effect of 1.5 groups is most obvious.

TABLE 1 influence of Chinese herbal immunopotentiator on growth and performance indexes of grouper

Weight gain (%) (final weight-initial weight) × 100/initial weight

Survival (%). Final Fish tails × 100/initial Fish tails

Feed coefficient is feed consumption/total fish weight gain

2) Influence of Chinese herbal medicine immunopotentiator on grouper intestinal flora

The influence of the Chinese herbal medicine immunopotentiator on the intestinal flora of the grouper is shown in figures 1-5, wherein 1.0 group, 1.5 group and 2.0 group are shown, the total number of bacteria, the number of lactic acid bacteria and the number of bifidobacteria in the intestinal tract of the grouper are obviously higher than those in a control group, and the number of pathogenic bacteria such as escherichia coli, vibrio and the like is obviously lower than that in the control group, so that the Chinese herbal medicine immunopotentiator can promote the proliferation of beneficial microorganisms (such as lactic acid bacteria and bifidobacteria) in the intestinal tract of the grouper, and improve the intestinal flora structure of the grouper by fighting for nutrients, competing for living space, secreting bacteriocin to inhibit the growth of harmful microorganisms and the like, thereby improving the disease resistance of the grouper.

3) Influence of Chinese herbal medicine immunopotentiator on immunity index of grouper

The Chinese herbal medicine immunopotentiator has immunity indexes for grouper as shown in fig. 6-9, including 1.0 group, 1.5 group and 2.0 group, the phagocytosis rate of leukocyte in blood of grouper is higher than that of control group and 0.5 group, and the activity of lysozyme, superoxide dismutase and antibacterial activity in blood serum are higher than that of control group. The group 1.5 was most effective.

4) Influence of Chinese herbal medicine immunopotentiator on disease resistance of grouper

The influence of the Chinese herbal medicines on the disease resistance of the groupers is shown in fig. 10, and the immune protection rates of the groupers in the test group are respectively as follows: 32.29% (0.5 group), 54.79% (1.0 group), 74.16% (1.5 group), 67.71% (2.0 group), and 1.5 group had the best disease resistance effect.

Immune protection (%). percent (control mortality-experimental mortality) x 100/control mortality

The results show that: 1.5 percent of Chinese herbal medicine immunopotentiator is added into the basic feed, the weight gain rate and the survival rate of the groupers in a test group are highest (respectively 44.94 percent and 8.33 percent higher than those in a control group), and the feed coefficient is lowest (0.39 percent lower than that in the control group); the immune indexes such as the phagocytic rate of leucocytes, the activity of superoxide dismutase, the activity of lysozyme, the antibacterial activity and the like in the blood of the grouper are obviously higher than those of a control group, the number of probiotics such as bifidobacteria, lactic acid bacteria and the like in the intestinal tract is obviously higher than that of the control group, and the number of pathogenic bacteria such as escherichia coli, vibrio and the like is obviously lower than that of the control group. After the grouper is infected with vibrio harveyi by artificial injection, the immune protection rate is 74.16%.

Test example 2

The Chinese herbal medicine immunopotentiator for the groupers in the examples 1 to 5 and the comparative examples 1 to 10 is added into grouper feed (commercial conventional grouper feed) according to the proportion of 1.5 percent (mass percentage), and is stirred and mixed uniformly to prepare the granular medicinal bait.

The prepared medicinal bait is used for feeding the margarita, gentian and grouper (grouper culture is carried out in a high-tech garden of Guangdong ocean university, east island of Zhanjiang city), the groups are respectively marked as an example 1-an example 5 and a comparative example 1-a comparative example 10, and the feeding management is carried out according to a scientific feeding method. The oxygen machine supplies oxygen, and water is changed for 1 time every two days. The daily feeding amount is 3-5% of the fish weight, and the feeding is carried out twice (9 am, 17 pm) in a feeding period of 28 days.

Weighing and counting fish in each group at 28 days, and taking 1 × 10 fish after 29 days⁷cells/mL of Vibrio harveyi bacterial suspension is subjected to toxicity attack, the injection dosage is 0.1 mL/tail, the death condition of each group of groupers within 10 days is observed and recorded, and the immune protection rate is calculated.

TABLE 2 influence of the herbal immunopotentiators of examples 1-5 and comparative examples 1-10 on various indexes of grouper

The results show that: the Chinese herbal medicine immunopotentiator for the grouper has the synergistic effect among the raw materials, particularly the prepared rehmannia root, the tuckahoe, the white atractylodes rhizome, the grassleaf sweelflag rhizome, the liquorice and the weeping forsythia. The Chinese herbal medicines are reasonably matched and used in the feed for the groupers, so that the weight gain rate, the survival rate, the feed coefficient and the immune protection rate of the groupers are superior to those of a comparative example, wherein the weight gain rate can reach 123.43%, the survival rate can reach 95%, the feed coefficient can be 0.67, and the immune protection rate can reach 74.16%, so that the survival rate and the disease resistance of the groupers are greatly improved.

The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.

Claims

1. The Chinese herbal medicine immunopotentiator for the groupers is characterized by comprising the following raw materials in parts by weight: 5-15 parts of prepared rehmannia root, 10-30 parts of mung bean, 5-20 parts of kudzu root, 1-10 parts of poria cocos, 1-10 parts of liquorice, 1-10 parts of radix curcumae, 1-10 parts of bighead atractylodes rhizome, 5-15 parts of rhizoma acori graminei, 1-10 parts of albizia flower, 1-10 parts of fructus forsythiae, 5-20 parts of dandelion, 5-10 parts of longan seed, 1-10 parts of costus root and 1-15 parts of hawthorn.

2. The Chinese herbal medicine immunopotentiator for grouper according to claim 1 is characterized by comprising the following raw materials in parts by weight: 8-10 parts of prepared rehmannia root, 15-20 parts of mung bean, 10-15 parts of kudzu root, 5-8 parts of poria cocos, 5-7 parts of liquorice, 4-6 parts of radix curcumae, 4-7 parts of bighead atractylodes rhizome, 6-10 parts of rhizoma acori graminei, 5-8 parts of albizia flower, 4-6 parts of fructus forsythiae, 8-12 parts of dandelion, 7-9 parts of longan seed, 7-9 parts of costus root and 8-12 parts of hawthorn.

3. A method for preparing the Chinese herbal immunopotentiator for grouper as claimed in claim 1 or 2, comprising the steps of: 14 Chinese herbal medicines of prepared rehmannia root, mung bean, kudzu root, poria with poria cocos, liquorice, radix curcumae, bighead atractylodes rhizome, rhizoma acori graminei, albizia flower, fructus forsythiae, dandelion, longan seed, elecampane and hawthorn are respectively crushed and uniformly mixed in proportion to obtain the Chinese herbal medicine immunopotentiator for grouper.

4. The method for preparing a Chinese herbal immunopotentiator for grouper according to claim 3, wherein the pulverized particle size is required to pass through a 100 mesh sieve.

5. Use of the Chinese herbal immunopotentiator for grouper as claimed in claim 1 or 2 as a feed additive.

6. The use of the Chinese herbal immunopotentiator for groupers as a feed additive according to claim 5, wherein the Chinese herbal immunopotentiator is added into the groupers feed, and the Chinese herbal immunopotentiator accounts for 0.5-2% of the mass of the groupers feed.