WO2021063047A1 - 黄芩地上部分的提取物在水产养殖中的应用 - Google Patents

黄芩地上部分的提取物在水产养殖中的应用 Download PDF

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WO2021063047A1
WO2021063047A1 PCT/CN2020/098625 CN2020098625W WO2021063047A1 WO 2021063047 A1 WO2021063047 A1 WO 2021063047A1 CN 2020098625 W CN2020098625 W CN 2020098625W WO 2021063047 A1 WO2021063047 A1 WO 2021063047A1
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scutellaria baicalensis
extract
parts
aerial part
feed
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PCT/CN2020/098625
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English (en)
French (fr)
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詹华强
董婷霞
夏翊腾
郑玉忠
汪成
戴坤
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香港科技大学
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/142—Amino acids; Derivatives thereof
    • A23K20/147—Polymeric derivatives, e.g. peptides or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/158—Fatty acids; Fats; Products containing oils or fats
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/539—Scutellaria (skullcap)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00—Treatment of water, waste water, or sewage
    • C02F1/50—Treatment of water, waste water, or sewage by addition or application of a germicide or by oligodynamic treatment
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish
    • Y02A40/818—Alternative feeds for fish, e.g. in aquacultures

Definitions

  • the invention relates to the technical field of aquaculture, in particular to the application of an extract of the aerial part of Scutellaria baicalensis Georgi in aquaculture.
  • the current research on antibacterial drugs mainly focuses on pathogenic bacteria that often appear in terrestrial environments, such as Staphylococcus, Streptococcus, Escherichia coli, Candida albicans, Pseudomonas aeruginosa, etc., and the targeted water-borne pathogens There is very little research.
  • the antibacterial methods used in aquaculture today are still based on copying human medicine and veterinary medicine, and there is a lack of precise research and medicine for aquatic microorganisms, which greatly reduces the efficiency of medicine and increases the cost of breeding.
  • the use of traditional chemicals and antibiotics can easily lead to drug resistance and multi-drug resistance of pathogens. Meat is located in the upper reaches of the human food chain, which will indirectly cause human resistance to antibiotics.
  • the residues of traditional chemicals and antibiotics in aquatic animals and the resulting distortion also seriously affect the value of aquatic economic animals.
  • one of the objectives of the present invention is to provide an application of an extract of the aerial part of Scutellaria baicalensis Georgi in aquaculture.
  • the extract of the aerial part of Scutellaria baicalensis has a good inhibitory effect on aquatic pathogens, and is not easy to produce drug resistance, and is suitable for large-scale application in aquaculture.
  • the aerial part of Scutellaria baicalensis Georgi includes at least one of Scutellaria baicalensis stem, Scutellaria baicalensis leaf and Scutellaria baicalensis flower.
  • Another object of the present invention is to provide an aquaculture feed, which is prepared from raw materials including the following parts by weight:
  • the extracts of aerial parts of Scutellariae are 0.5-5 parts, 90-110 parts of feed and 0.2-2 parts of fish oil.
  • Another object of the present invention is to provide a method for preparing the above-mentioned aquaculture feed, which includes the following steps:
  • the present invention has the following beneficial effects:
  • Scutellaria baicalensis is the root of Scutellaria baicalnsis Georgi. It tastes bitter and cold in nature. It has the effects of clearing heat and dampness, purging fire and detoxification. It is a commonly used Chinese medicinal material. It is planted in large areas in Heilongjiang, Liaoning, Inner Mongolia, Hebei, Henan, Gansu, Shaanxi, Shanxi, Shandong, Sichuan and other places. However, only the roots are used as medicine, and the rest of the stems and leaves are discarded as waste, which pollutes the environment and also causes a great waste of resources.
  • the inventors of the present invention have discovered through a large number of experimental studies that the extract of the aerial part of Scutellaria baicalensis Georgi has a good inhibitory effect on aquatic pathogens and is not easy to produce drug resistance, so it is suitable for aquaculture.
  • the aquaculture feed prepared by compounding the extract of the aerial part of Scutellaria baicalensis Georgi with fish oil and feed in a specific ratio has a significant inhibitory effect on microorganisms in the water body, and has a good anti-inflammatory effect on aquatic animals.
  • the addition of fish oil is avoided.
  • the preparation process of the aquaculture feed of the invention is simple, realizes the high value-added utilization of the above-ground part of the traditional Chinese medicine waste Scutellaria baicalensis Georgi, greatly reduces the cost of raw materials, and is suitable for large-scale production.
  • Figure 1 is the preparation process of aquaculture feed for the aerial part of Scutellaria baicalensis Georgi in Example 1;
  • Figure 2 shows the inhibitory effect of the extract of the aerial part of Scutellaria baicalensis Georgi on the growth of aquatic bacteria in Example 4;
  • Figure 3 shows the in vitro anti-inflammatory results of the extract of the aerial part of Scutellaria baicalensis Georgi in Example 6;
  • Figure 4 shows the in vivo antibacterial and anti-inflammatory results of aquatic feed supplemented with extracts of aerial parts of Scutellaria baicalensis in Example 7 (* represents a significant difference compared with the blank group, P ⁇ 0.05; ** represents a very significant difference compared with the blank group, P ⁇ 0.01);
  • Fig. 5 shows the in vivo antibacterial results of aquatic feed supplemented with extracts of aerial parts of Scutellaria baicalensis Georgi in Example 7. Figs.
  • the embodiment of the present invention provides the application of the extract of the aerial part of Scutellaria baicalensis Georgi in aquaculture; the aerial part of Scutellaria baicalensis Georgi includes at least one of scutellaria scutellaria stem, scutellaria baicalensis leaf, and scutellaria baicalensis flower.
  • the extract of the aerial part of Scutellaria baicalensis Georgi includes an alcohol extract, a water extract or an aqueous ethanol extract of the aerial part of Scutellaria baicalensis Georgi or a refined product thereof.
  • the extract of the aerial part of Scutellaria baicalensis Georgi is used in the preparation of aquaculture feed.
  • the extract of the aerial part of Scutellaria baicalensis Georgi is used as an additive for the aquaculture feed.
  • antibiotics are not included in the aquaculture feed.
  • the preparation of the extract of the aerial part of Scutellaria baicalensis Georgi includes the following steps: extracting the aerial part of Scutellaria baicalensis Georgi with an extraction solvent, concentrating, and drying.
  • the preparation of the extract of the aerial part of Scutellariae includes the following steps: take the powder of the aerial part of Scutellariae, add 12-18 times (W/V) of the extraction solvent, sonicate at room temperature for 20-40 minutes, and centrifuge , Collect the supernatant, repeat the above steps once for the obtained precipitate, combine the supernatants to obtain the extract of the aerial part of Scutellaria baicalensis Georgi, concentrate and dry.
  • the preparation of the refined product of the alcohol extract, water extract, or ethanol aqueous extract of the aerial part of Scutellariae includes the following steps: extracting the aerial part of Scutellariae with an extraction solvent, concentrating the resulting extract, and adjusting The pH is 2-3, separated with a macroporous resin, and the eluted portion of the ethanol aqueous solution with a volume fraction of 80-90% is collected and concentrated.
  • the extraction solvent includes water, ethanol, or ethanol aqueous solution.
  • the volume fraction of ethanol in the ethanol aqueous solution is 70-80%.
  • the extraction method is ultrasonic extraction or thermal extraction
  • the embodiment of the present invention also provides an aquaculture feed, which is prepared from raw materials including the following parts by weight:
  • the extracts of aerial parts of Scutellariae are 0.5-5 parts, 90-110 parts of feed and 0.2-2 parts of fish oil.
  • the aquaculture feed is prepared from raw materials including the following parts by weight: 1.5-2.5 parts of aerial part extract of Scutellaria baicalensis Georgi, 95-105 parts of feed, and 0.2-1 part of fish oil.
  • the feed in the raw material includes the following components in parts by weight: 30-40 parts by weight of protein, 6-7 parts by lipid, 3-6 parts by cellulose, and 29-37 parts by carbohydrate.
  • the feed in the raw material includes the following components in parts by weight: 30-40 parts by weight of protein, 6-7 parts of lipids, 3-6 parts of cellulose, 29-37 parts of carbohydrates, and moisture. 8-9 servings.
  • the above-mentioned preparation method of aquaculture feed includes the following steps:
  • the method for preparing the above-mentioned aquaculture feed includes the following steps:
  • aerial parts of Scutellaria refer to stems, leaves, flowers, etc. other than the roots of Scutellaria baicalensis.
  • the extraction solvent in step (1) includes water, ethanol, or ethanol aqueous solution.
  • the extraction method in step (1) is ultrasonic extraction or thermal extraction.
  • the ultrasound time is 20-40 min.
  • the mass volume concentration of the aqueous solution of the extract of the aerial part of Scutellaria baicalensis Georgi in step (2) is 50-150 mg/ml.
  • the mass volume concentration of the aqueous solution of the aerial part of Scutellaria baicalensis Georgi is 90-110 mg/ml.
  • the drying temperature in step (2) is 35-60°C.
  • Raw materials aerial parts of Scutellaria baicalensis (including Scutellaria baicalensis stems, leaves or flowers), fish oil, feed (30-40 parts of protein, 6-7 parts of lipids, 3-6 parts of cellulose, 29-37 parts of carbohydrates and 8-9 parts of moisture Copies).
  • microorganisms involved in the following examples (Aeromonas hydrophila, Edwards lenta, Vibrio alginolyticus, Vibrio harveyi, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa ) Were purchased from Guangdongzhou Microbial Culture Collection.
  • the SGT method was used to evaluate the antibacterial activity of the extract of the aerial part of Scutellaria baicalensis Georgi.
  • the initial growth curve method (SGT) is a new high-throughput screening method for screening antimicrobial active drugs. After the bacteria are co-cultured with the drug for a period of time, the drug is removed by centrifugation, and the remaining bacteria are diluted into a new medium for growth. The growth status of the bacteria is evaluated by reading the OD595. After a period of time, it enters the logarithmic growth phase. Bacteria enter the logarithmic growth phase faster, and fewer bacteria enter the logarithmic growth phase slower. By calculating the time points when different groups enter the logarithmic growth phase, and then calculating the number of bacteria after co-cultivation with the drug, the antibacterial activity of the drug can be judged.
  • aqueous extracts and alcohol extracts of the aerial parts of Scutellaria baicalensis prepared in Examples 1 and 2 were dissolved into a solution with a final concentration of 100 mg/mL, and further configured into a solution of 10 mg/mL and 1 mg/mL with four water-borne pathogens, respectively.
  • kanamycin was prepared with DMSO to a concentration of 5mg/mL as a control.
  • the refined water extract of the aerial part of Scutellariae prepared in Example 3 was dissolved in dimethyl sulfoxide (DMSO) to prepare a solution with a final concentration of 1 mg/mL, and further diluted with cell culture solution to a concentration of 0.05 respectively. mg/mL, 0.1mg/mL, 0.15mg/mL, 0.2mg/mL solutions, respectively, co-culture with macrophages;
  • DMSO dimethyl sulfoxide
  • the extract of the aerial part of Scutellaria baicalensis Georgi has a significant inhibitory effect on the expression of inflammatory transcription factor NF- ⁇ B and inflammatory cytokine IL-1 ⁇ in macrophages, and has good anti-inflammatory activity in vitro. Its anti-inflammatory effect has also increased, and it is suitable for being widely used as aquatic feed additives.
  • Feed the aquaculture feed prepared in Example 1 to the experimental mud meng fish feeding amount: each fish is fed about 2.5 grams of feed per day, once in the morning and once in the evening), and at the same time add four mixed bacteria to the culture In the water tank.
  • the feed test result without adding any extract was used as the negative control of the experiment.
  • the test results of feed supplemented with antibiotics such as enrofloxacin added amount: final concentration of enrofloxacin in aquaculture water: 0.05 mg/mL) were used as a positive control for the experiment.
  • the test result without adding bacteria was used as the experimental blank control.
  • the aquaculture feed added with the extract of the aerial part of Scutellaria baicalensis according to the present invention can reduce the expression of inflammatory factors in various tissues of Scutellaria baicalensis and reduce the number of bacteria in the aquaculture water.

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Abstract

本发明涉及一种黄芩地上部分的提取物在水产养殖中的应用。所述黄芩地上部分包括黄芩茎、黄芩叶和黄芩花中的至少一种。其中，黄芩地上部分的提取物对于水生病原菌具有良好的抑制效果，且不易产生耐药性，适合于在水产养殖中大规模应用。

Description

黄芩地上部分的提取物在水产养殖中的应用 技术领域
本发明涉及水产养殖技术领域，特别是涉及一种黄芩地上部分的提取物在水产养殖中的应用。
背景技术
近年来，随着水产养殖规模的不断扩大、养殖种类的增加以及环境的日益恶化，各类水产养殖病害呈频发的态势，其中水生病原菌引起的水产动物细菌性疾病，仍然是最常见、危害最大的疾病之一。
但是，一方面，目前针对抗菌药物的研究主要针对陆地环境中常出现的致病菌，如葡萄球菌、链球菌、大肠杆菌、白色念珠菌、铜绿假单胞菌等，而对于水生病原菌的针对性研究十分少。并且现今水产养殖中的抗菌手段仍然以照搬人医、兽医用药为主，缺乏针对水产微生物的精准研究和用药，大大降低了用药效率，提高了养殖成本。另一方面，传统化学药品以及抗生素的使用，容易导致病原菌的耐药性以及多重耐药性。而肉类位于人类食物链的上游，会间接造成人类对抗生素的抗药性。并且传统化学药品以及抗生素在水产动物体内的残留及由此产生的畸变也严重影响了水产经济动物的价值。
因此，如何提供一种具有良好的抗水生病原菌功效、低毒且不易产生耐药性的药物应用于水产养殖，具有重要意义。
发明内容
基于此，本发明的目的之一是提供一种黄芩地上部分的提取物在水产养殖中的应用。该黄芩地上部分的提取物对于水生病原菌具有良好的抑制效果，且不易产生耐药性，适合于在水产养殖中大规模应用。
具体技术方案如下：
黄芩地上部分的提取物在水产养殖中的应用；所述黄芩地上部分包括黄芩茎、黄芩叶和黄芩花中的至少一种。
本发明的另一目的是提供一种水产养殖饲料，由包括以下重量份的原料制备而成：
黄芩地上部分的提取物0.5-5份、饲料90-110份和鱼油0.2-2份。
本发明的再一目的是提供上述水产养殖饲料的制备方法，包括以下步骤：
(1)黄芩地上部分的提取物的制备：将黄芩地上部分用提取溶剂提取，浓缩，干燥，得黄芩地上部分的提取物；
(2)按所述重量份数称取黄芩地上部分的提取物，溶于水后喷洒在所述饲料的表面，干燥；
(3)在步骤(2)得到的饲料表面喷洒所述鱼油。
与现有技术相比，本发明具有以下有益效果：
黄芩是唇形科植物(Scutellaria baicalnsis Georgi)的根，味苦性寒，具有清热燥湿，泻火解毒的功效，是常用中药材。在黑龙江、辽宁、内蒙古、河北、河南、甘肃、陕西、山西、山东、四川等地均有大面积种植。但仅只有根部用来入药，其余茎叶均作为废弃物丢弃，污染环境的同时还造成资源的极大浪费。
本发明的发明人经过大量的试验研究发现，黄芩地上部分的提取物对于水生病原菌具有良好的抑制效果，且不易产生耐药性，适合用于水产养殖中。
将黄芩地上部分的提取物与鱼油、饲料按特定比例复配，制备得到的水产养殖饲料，对于水体中的微生物具有明显的抑制作用，对于水产动物具有良好的抗炎作用，同时鱼油的添加避免了黄芩味苦造成的水产动物食用困难的问题。
本发明的水产养殖饲料制备工艺简单，实现了中药废弃物黄芩的地上部分作的高附值利用，大大降低了原料成本，可适用于大规模生产。
附图说明
图1为实施例1中黄芩地上部分的水产养殖饲料制备工艺；
图2为实施例4中黄芩地上部分的提取物对水生细菌的生长抑制作用；
图3是实施例6中黄芩地上部分的提取物的体外抗炎结果；
图4是实施例7中添加黄芩地上部分的提取物的水产饲料的体内抗菌抗炎 结果(*代表与空白组比较有显著差异，P<0.05；**代表与空白组比较有极显著差异，P<0.01)；
图5是实施例7中添加黄芩地上部分的提取物的水产饲料的体内抗菌结果。
具体实施方式
为了便于理解本发明，下面将参照实施例对本发明进行更全面的描述，以下给出了本发明的较佳实施例。但是，本发明可以以许多不同的形式来实现，并不限于本文所描述的实施例。提供这些实施例的目的是使对本发明的公开内容的理解更加透彻全面。
除非另有定义，本文所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。在本发明的说明书中所使用的术语只是为了描述具体的实施例的目的，不是旨在于限制本发明。本文所使用的术语“和/或”包括一个或多个相关的所列项目的任意的和所有的组合。
本发明的实施例提供黄芩地上部分的提取物在水产养殖中的应用；所述黄芩地上部分包括黄芩茎、黄芩叶和黄芩花中的至少一种。
在其中一些实施例中，所述黄芩地上部分的提取物包括所述黄芩地上部分的醇提取物、水提取物或乙醇水溶液提取物或其精制品。
在其中一些实施例中，黄芩地上部分的提取物在制备水产养殖饲料中的应用。
在其中一些实施例中，黄芩地上部分的提取物作为所述水产养殖饲料的添加剂。
在其中一些实施例中，所述水产养殖饲料中不包括抗生素。
在其中一些实施例中，所述黄芩地上部分的提取物的制备包括以下步骤：将黄芩地上部分用提取溶剂提取，浓缩，干燥。
在其中一些实施例中，所述黄芩地上部分的提取物的制备包括以下步骤：取黄芩地上部分粉末，加入其12-18倍(W/V)的提取溶剂，室温超声20-40分钟，离心，收集上清液，所得沉淀重复上述步骤一次，合并上清液，得到黄芩地上部分的提取液，浓缩，干燥。
在其中一些实施例中，所述黄芩地上部分的醇提取物、水提取物或乙醇水溶液提取物的精制品的制备包括以下步骤：将黄芩地上部分用提取溶剂提取，将所得提取液浓缩，调节pH至2-3，用大孔树脂分离，收集体积分数为80-90％的乙醇水溶液洗脱部分，浓缩。
在其中一些实施例中，所述提取溶剂包括水、乙醇或乙醇水溶液。优选地，所述乙醇水溶液中乙醇的体积分数为70-80％。
在其中一些实施例中，所述提取的方式为超声提取或热提取；
本发明的实施例还提供一种水产养殖饲料，由包括以下重量份的原料制备而成：
黄芩地上部分的提取物0.5-5份、饲料90-110份和鱼油0.2-2份。在其中一些实施例中，所述水产养殖饲料由包括以下重量份的原料制备而成：黄芩地上部分的提取物1.5-2.5份、饲料95-105份、鱼油0.2-1份。
在其中一些实施例中，所述原料中的饲料包括以下重量份数的组分：蛋白质30-40份、脂类6-7份、纤维素3-6份和碳水化合物29-37份。
在其中一些实施例中，所述原料中的饲料包括以下重量份数的组分：蛋白质30-40份、脂类6-7份、纤维素3-6份、碳水化合物29-37份和水分8-9份。
上述水产养殖饲料的制备方法，包括以下步骤：
(1)黄芩地上部分的提取物的制备：将黄芩地上部分用提取溶剂提取，浓缩，干燥，得黄芩地上部分的提取物；
(2)按所述重量份数称取黄芩地上部分的提取物，溶于水后喷洒在所述饲料的表面，干燥；
(3)在步骤(2)得到的饲料表面喷洒所述鱼油。
在其中一些实施例中，上述水产养殖饲料的制备方法，包括以下步骤：
(1)黄芩地上部分的提取物的制备：将黄芩地上部分用提取溶剂提取，将所得提取液浓缩，调节pH至2-3，用大孔树脂分离，收集体积分数为80-90％的乙醇水溶液洗脱部分，浓缩，得黄芩地上部分的提取物；
(2)按所述重量份数称取黄芩地上部分的提取物，溶于水后喷洒在所述饲料的表面，干燥；
(3)在步骤(2)得到的饲料表面喷洒所述鱼油。
其中，所述黄芩地上部分指的是除黄芩根之外的茎、叶、花等。
在其中一些实施例中，步骤(1)所述提取溶剂包括水、乙醇或乙醇水溶液。
在其中一些实施例中，步骤(1)所述提取的方式为超声提取或热提取。优选地，所述超声的时间为20-40min。
在其中一些实施例中，步骤(2)所述的黄芩地上部分的提取物的水溶液的质量体积浓度为50-150mg/ml。优选地，所述的黄芩地上部分的提取物的水溶液的质量体积浓度为90-110mg/ml。
在其中一些实施例中，步骤(2)所述干燥的温度为35-60℃。
以下结合具体实施例对本发明作进一步详细的说明。
原料：黄芩地上部分(包括黄芩茎、叶或花)、鱼油、饲料(蛋白质30-40份、脂类6-7份、纤维素3-6份、碳水化合物29-37份和水分8-9份)。
以下实施例涉及的微生物(嗜水气单胞菌、迟缓爱德华氏菌、解藻弧菌、哈维氏弧菌、金黄色葡萄球菌、大肠杆菌、肺炎克雷伯氏菌、铜绿假单胞菌)均购买自广东省微生物菌种保藏中心。
实施例1 水产养殖饲料的制备方法
包括以下步骤：
(1)黄芩地上部分的水提物的制备：取黄芩地上部分粉末，加入15倍体积的水，室温超声30分钟，4000rpm离心10分钟后，收集上清液，沉淀重复上述步骤一次，合并上清液，得到黄芩地上部分提取液。将黄芩地上部分提取液减压浓缩，冷冻干燥得到黄芩地上部分水提物粉末
(2)将黄芩地上部分水提物粉末加水溶解，配成终浓度为100mg/mL；将配方水产饲料(颗粒直径0.2cm)放入饲料搅拌机；按饲料干重的2％比例(粉末质量)喷洒黄芩地上部分的水提物的水溶液，以40转/分钟的转速搅拌，使黄芩地上部分的水提物的水溶液均匀包裹在饲料表面；
(3)45℃低温烘干饲料，按饲料干重的0.5％比例喷洒加入鱼油，40转/分钟搅拌，使鱼油均匀包裹在饲料表面上，即得水产养殖饲料。
实施例2
(1)黄芩地上部分的醇提物的制备：以黄芩地上部分粉末加入15倍体积的75％浓度的乙醇，室温超声30分钟，4000rpm离心10分钟后，收集上清液，所得沉淀重复上述步骤一次，合并上清液，得到黄芩地上部分提取液。将黄芩地上部分提取液减压浓缩，冷冻干燥得到黄芩地上部分醇提物粉末。
(2)将黄芩地上部分醇提物粉末加水溶解，配成终浓度为100mg/mL；将配方水产饲料(颗粒直径0.2cm)放入饲料搅拌机；按饲料干重的5％比例(粉末质量)喷洒黄芩地上部分的提取物的水溶液，以40转/分钟的转速搅拌，使黄芩地上部分的提取物水溶液均匀包裹在饲料表面；
(3)45℃低温烘干饲料，按饲料干重的1％比例喷洒加入鱼油，40转/分钟搅拌，使鱼油均匀包裹在饲料表面上，即得水产养殖饲料。
实施例3
(1)黄芩地上部分的水提取物的精制品的制备：以黄芩地上部分粉末加入15倍体积的水，室温浸泡30分钟后，加热提取2小时，收取上清液并加热浓缩，调PH至2-3，上大孔树脂柱，收85％乙醇洗脱部分，减压浓缩即得。
(2)将黄芩地上部分的水提取物的精制品粉末加水溶解，配成终浓度为100mg/mL；将配方水产饲料(颗粒直径0.2cm)放入饲料搅拌机；按饲料干重的1％比例(粉末质量)喷洒黄芩地上部分的提取物水溶液，以40转/分钟的转速搅拌，使黄芩地上部分的提取物水溶液均匀包裹在饲料表面；
(3)35℃低温烘干饲料，按饲料干重的0.2％比例喷洒加入鱼油，40转/分钟搅拌，使鱼油均匀包裹在饲料表面上，即得水产养殖饲料。
实施例4 体外抗菌活性检测
1、将实施例1和2制备得到的黄芩地上部分提取物，溶解配成终浓度为100mg/mL的溶液，进一步配置成浓度为10mg/mL、1mg/mL溶液分别与嗜水气单胞菌、迟缓爱德华氏菌、解藻弧菌和哈维氏弧菌四种有害细菌的菌液共培养12 小时；
2、培养结束后离心去除旧培养基，将菌体用新培养基稀释后测量生长曲线；
3、采用SGT法评估黄芩地上部分的提取物的抗菌活性。起始生长曲线法(SGT)是筛选抗菌活性药物的高通量新筛选方法。细菌在与药物共培养一段时间后，离心去除药物，剩余细菌稀释到新的培养基中进行生长，通过读取OD595评估细菌生长状态，一段时间后进入对数生长期，起始数量较多的细菌较快进入对数生长期，较少的细菌较慢进入对数生长期。通过计算不同组进入对数生长期的时间点，进而计算与药物共培养后细菌的数量，从而判断药物的抗菌活性。
如附图2所示，黄芩地上部分的不同提取液对四种水生有害细菌均有抑制作用。由此可知，黄芩地上部分的提取物对于水生病原菌具有良好的生长抑制作用。
实施例5 体外抗菌活性检测
将实施例1和2制备得到的黄芩地上部分水提物和醇提物，溶解配成终浓度为100mg/mL的溶液，进一步配置成10mg/mL、1mg/mL的溶液分别与四种水生病原菌(嗜水气单胞菌、迟缓爱德华氏菌、解藻弧菌和哈维氏弧菌)和四种常见非水生病原菌(金黄色葡萄球菌、大肠杆菌、肺炎克雷伯氏菌、铜绿假单胞菌)的菌液共培养12小时；卡那霉素用DMSO配置成浓度为5mg/mL的溶液，作为对照。
结果如表1所示
表1
Figure PCTCN2020098625-appb-000001
Figure PCTCN2020098625-appb-000002
由表1的结果可知，黄芩地上部分提取物在高浓度(10mg/mL)对四种水生病原菌(嗜水气单胞菌、迟缓爱德华氏菌、解藻弧菌和哈维氏弧菌)和四种常见非水生病原菌(金黄色葡萄球菌、大肠杆菌、肺炎克雷伯氏菌、铜绿假单胞菌)的抑菌功效明显。
实施例6 体外抗炎活性检测
1、将实施例3制备得到的黄芩地上部分的水提物精制品用二甲基亚砜(DMSO)溶解配成终浓度为1mg/mL的溶液，用细胞培养液进一步稀释成浓度为分别0.05mg/mL、0.1mg/mL、0.15mg/mL、0.2mg/mL的溶液，分别与巨噬细胞共同培养；
2、以脂多糖刺激巨噬细胞的炎症反应，同时以没有加入任何的对照的测试结果作为实验阴性对照，以抗炎药物地塞米松的测试结果作为实验阳性对照；
3、通过荧光素酶法及实时定量PCR法测量炎症转录因子NF-κB及炎症细胞因子IL-1β的表达量。
如附图3所示，黄芩地上部分的提取物对巨噬细胞中炎症转录因子NF-κB以及炎症细胞因子IL-1β的表达有明显抑制作用，具有良好的体外抗炎活性，且随着浓度的提升其抗炎效果也有所增加，适合广泛用于水产饲料添加剂。
实施例7 体内抗菌、抗炎实验
1、将实施例1制备的水产养殖饲料投喂给试验用泥猛鱼(投喂量为：每条鱼每日投喂大约2.5克饲料，早晚各一次)，同时加入四种混合细菌到养殖水箱中。以没有加入任何提取物的饲料测试结果作为实验阴性对照。以添加恩诺沙星等抗生素的饲料(添加量为：恩诺沙星在养殖水体中最终浓度：0.05mg/mL) 测试结果作为实验阳性对照。以不添加细菌的测试结果作为实验空白对照。
2、通过梯度稀释法测量养殖过程中水体中细菌的数量变化；
3、比较加入细菌培养48小时后鱼内各个组织内先炎症因子的表达量。
如附图4和附图5所示，本发明添加了黄芩地上部分的提取物的水产养殖饲料可以减少泥猛鱼各组织内炎症因子的表达，降低养殖水体中细菌的数量。
以上所述实施例的各技术特征可以进行任意的组合，为使描述简洁，未对上述实施例中的各个技术特征所有可能的组合都进行描述，然而，只要这些技术特征的组合不存在矛盾，都应当认为是本说明书记载的范围。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。

Claims (10)

  1. 黄芩地上部分的提取物在水产养殖中的应用；所述黄芩地上部分包括黄芩茎、黄芩叶和黄芩花中的至少一种。
  2. 根据权利要求1所述的应用，其特征在于，所述黄芩地上部分的提取物包括所述黄芩地上部分的醇提取物、水提取物或乙醇水溶液提取物或其精制品。
  3. 根据权利要求1所述的应用，其特征在于，黄芩地上部分的提取物在制备水产养殖饲料中的应用。
  4. 根据权利要求3所述的应用，其特征在于，黄芩地上部分的提取物作为所述水产养殖饲料的添加剂。
  5. 一种水产养殖饲料，其特征在于，由包括以下重量份的原料制备而成：
    权利要求1-8任一项应用中所述的黄芩地上部分的提取物0.5-5份、饲料90-110份和鱼油0.2-2份。
  6. 根据权利要求5所述的水产养殖饲料，其特征在于，由包括以下重量份的原料制备而成：
    黄芩地上部分的提取物1.5-2.5份、饲料95-105份和鱼油0.2-1份。
  7. 根据权利要求5所述的水产养殖饲料，其特征在于，所述原料中的饲料包括以下重量份数的组分：
    蛋白质30-40份、脂类6-7份、纤维素3-6份和碳水化合物29-37份。
  8. 权利要求5-7任一项所述水产养殖饲料的制备方法，其特征在于，包括以下步骤：
    (1)黄芩地上部分的提取物的制备：将黄芩地上部分用提取溶剂提取，浓缩，干燥，得黄芩地上部分的提取物；
    (2)按所述重量份数称取黄芩地上部分的提取物，溶于水后喷洒在所述饲料的表面，干燥；
    (3)在步骤(2)得到的饲料表面喷洒所述鱼油。
  9. 根据权利要求8所述的制备方法，其特征在于，步骤(2)所述黄芩地上部分的提取物的水溶液的质量体积浓度为(50-150)mg/ml。
  10. 根据权利要求8或9所述的制备方法，其特征在于，步骤(2)所述干燥的温度为35-60℃。
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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101433272A (zh) * 2008-12-26 2009-05-20 福建大昌生物科技实业有限公司 水产高能饲料制造工艺
KR20140055882A (ko) * 2012-11-01 2014-05-09 한경옥 온실가스 저감용 친환경 반추동물용 가축사료
CN105325684A (zh) * 2015-10-15 2016-02-17 张厚冰 一种促进虾蛋白质合成的南瓜渣饲料及其制备方法
CN105901328A (zh) * 2016-05-18 2016-08-31 中国水产科学研究院淡水渔业研究中心 异育银鲫复合中草药抗嗜水气单胞菌制剂及其制备方法和应用
CN106962684A (zh) * 2017-04-14 2017-07-21 福建高农饲料有限公司 欧洲鳗专用膨化饲料及其制备方法
CN109221649A (zh) * 2018-09-03 2019-01-18 汕头大学 一种提高双棘黄姑鱼鱼鳔品质的人工配合饲料及其制备
CN109744403A (zh) * 2017-11-05 2019-05-14 季雪松 一种鲦鱼类专用膨化饲料的加工工艺
CN110548066A (zh) * 2019-09-30 2019-12-10 香港科技大学 黄芩地上部分的提取物在水产养殖中的应用

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1131066C (zh) * 1998-12-31 2003-12-17 吉林省东北亚药业股份有限公司 黄芩茎叶在制备镇痛、抗炎或／和解热药物中的应用
CN104352617A (zh) * 2014-10-11 2015-02-18 中国科学院海洋研究所 一种三黄散超微粉及其制备方法和应用
CN105168667B (zh) * 2015-10-27 2020-07-07 上海海洋大学 中草药在延缓细菌耐药性中的应用
CN106721625A (zh) * 2016-12-07 2017-05-31 重庆文洪水产品养殖有限公司 一种抗氧化水产沉性饲料及其制备方法

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101433272A (zh) * 2008-12-26 2009-05-20 福建大昌生物科技实业有限公司 水产高能饲料制造工艺
KR20140055882A (ko) * 2012-11-01 2014-05-09 한경옥 온실가스 저감용 친환경 반추동물용 가축사료
CN105325684A (zh) * 2015-10-15 2016-02-17 张厚冰 一种促进虾蛋白质合成的南瓜渣饲料及其制备方法
CN105901328A (zh) * 2016-05-18 2016-08-31 中国水产科学研究院淡水渔业研究中心 异育银鲫复合中草药抗嗜水气单胞菌制剂及其制备方法和应用
CN106962684A (zh) * 2017-04-14 2017-07-21 福建高农饲料有限公司 欧洲鳗专用膨化饲料及其制备方法
CN109744403A (zh) * 2017-11-05 2019-05-14 季雪松 一种鲦鱼类专用膨化饲料的加工工艺
CN109221649A (zh) * 2018-09-03 2019-01-18 汕头大学 一种提高双棘黄姑鱼鱼鳔品质的人工配合饲料及其制备
CN110548066A (zh) * 2019-09-30 2019-12-10 香港科技大学 黄芩地上部分的提取物在水产养殖中的应用

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