WO2019100540A1 - 一种南美白对虾饲料添加剂的制备方法 - Google Patents

一种南美白对虾饲料添加剂的制备方法 Download PDF

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WO2019100540A1
WO2019100540A1 PCT/CN2018/000043 CN2018000043W WO2019100540A1 WO 2019100540 A1 WO2019100540 A1 WO 2019100540A1 CN 2018000043 W CN2018000043 W CN 2018000043W WO 2019100540 A1 WO2019100540 A1 WO 2019100540A1
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fermentation
feed additive
parts
fermentation broth
malt
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PCT/CN2018/000043
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English (en)
French (fr)
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陈程
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陈程
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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
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • 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
    • A23K20/174—Vitamins
    • 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
    • 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
    • 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
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

Definitions

  • the invention belongs to the technical field of preparation of feed additives for aquaculture, and particularly relates to a preparation method of a feed additive for South American shrimp.
  • Penaeus vannamei belongs to the prawn family, and its scientific name is prawn, which is one of the top three species of shrimp produced in the world.
  • Penaeus vannamei has large individual body and thin shell body fat.
  • the meat yield is as high as 65%.
  • the meat is fresh and tender, the taste is delicious, and the protein content is rich. It is very popular in domestic and foreign markets and has high economic value.
  • the artificial breeding of white shrimp in South America is very The broad range is the pillar industry of marine aquaculture in China. In addition to the need for good growth environment and scientific management, Penaeus vannamei also needs balanced feed.
  • Chinese herbal medicine has the advantages of less residue and less resistance to pathogenic bacteria. Its function not only has direct inhibition and killing effect on pathogenic microorganisms, but more importantly, it adjusts the immune system of feeding animals and improves the feeding of animals. The immune function and anti-stress ability, the Chinese herbal medicine ingredients are made into feed additives, and the diseases that are easy to occur in the control of white shrimp culture in South America are in full compliance with the disease prevention and control guidelines for the development of pollution-free aquatic industry and production of green aquatic products. In addition, Chinese herbal medicines have a variety of nutrients and bioactive substances. Choosing appropriate Chinese herbal medicine varieties can also promote the growth of P. vannamei and improve the quality of prawns.
  • the invention patent CN201310293138.0 is a special compound feed for Penaeus vannamei, which is composed of the following components by weight: 18-24% of steam fish meal, 17-22% of soybean meal, 15-24% of peanut meal, and 2-4 of blood globulin powder. %, squid cream 2 ⁇ 4%, yeast cream 2-5%, flour 20-25%, fish oil 2-3%, soybean oil 1-2%, shrimp multi-dimensional 1.0%, shrimp multi-mine 2.0%, South American white shrimp special feed The additive was 1.0%, and the sum of the above components was 100%.
  • the nutritional composition of the compound feed of the invention is determined by repeated experiments according to the nutritional requirements of Penaeus vannamei, which can effectively meet the nutritional needs of Penaeus vannamei, improve the immunity and disease resistance of the shrimp, and promote the healthy growth of the shrimp. .
  • Invention patent CN201210307270.8 A kind of Chinese herbal compound feed additive for attracting white prawn, from 15 to 20% hawthorn, 15 to 20% rhubarb, 10 to 18% jaundice, 10 to 14% licorice, 10 to 15% Wei, 6 to 8% earthworm, 5 to 6% divine, 3 to 5% amomum, 3 to 5% dried ginger, 4 to 5% taurine, 4 to 6% betaine, taurine and betaine Compounded.
  • the feed additive of the invention can have a significant attracting effect on P. vannamei, and can replace the increasingly tight animal raw materials (especially squid paste) to meet the development needs of the P. vannamei industry.
  • Invention patent CN201210307336.3 A Chinese herbal medicine feed additive for preventing enteritis of Penaeus vannamei, which is prepared by mixing gentian, phellodendron, tangerine peel, magnolia, rhubarb, medlar, astragalus and astragalus, together with corncob powder.
  • the feed additive of the invention can be used for preventing enteritis of Penaeus vannamei, and has the advantages of increasing the growth rate of Penaeus vannamei, enhancing the immunity of the shrimp, and improving the success rate of the culture.
  • the industry mainly uses the microecological preparations to regulate the water body of P. vannamei to prevent the intestinal inflammatory disease of P. vannamei, but because the quality of many products of microecological preparation manufacturers is uneven, leading to enteritis after the actual breeding process The prevention effect is not good, which brings certain breeding risks and cost waste to the farmers.
  • the invention provides a preparation method of a white prawn feed additive.
  • a preparation method of a white prawn feed additive is prepared by the following steps:
  • the composite bacteria I is a combination of 4 to 6% of Bacillus natto bacteria and 3 to 5% of Lactobacillus rhamnosus.
  • the composite bacteria II is a combination of 4 to 6% of Bacillus subtilis Bacillus subtilis and 2 to 4% of Aspergillus niger.
  • the precipitate obtained by the first centrifugation may be added to the double-fold amount of distilled water for washing, and then the precipitate is removed by centrifugation, and the obtained filtrate is separated from the filtrate by the first centrifugation. merge;
  • the grapefruit peel of the step (1) is dried to a moisture content of 10 to 20% and pulverized to 15 to 20 mesh.
  • the particle size affects the leaching rate of folic acid, and the folic acid can be completely and preferentially extracted and leached in 15-20 mesh.
  • the water temperature of the malt and water immersion in the step (1) is 15 to 20 ° C
  • the specific fermentation method is as follows: the mashed malt is added to the water of 1 to 1.5 times the mass and 1 to 1.2% of the mass.
  • Sodium chloride was inoculated with 3% yeast at 35 ° C for 6 h to obtain a malt fermentation broth.
  • the phytic acid in malt gradually decreases with the growth of malt, the phytic acid activity decreases at low temperature, the malt growth is inhibited, and the consumption of phytic acid is reduced.
  • the yeast fermentation changes the structure and metabolism of malt and increases the yield of phytic acid.
  • Grapefruit contains folic acid, which promotes gastrointestinal amino acid and sugar metabolism, enhances choline synthesis, and promotes fat metabolism.
  • Folic acid also promotes the decomposition of rapeseed cake and camellia cake to release phytic acid and tea saponin; malt and rapeseed meal It is rich in phytic acid, which can promote the fermentation efficiency of bacteria and increase the enzyme content.
  • Phosphorus after phytic acid hydrolysis can increase the enzyme activity of the fermentation product, thereby improving the nutritional value of the enzyme; natural bentonite can effectively adsorb the protein in the extract.
  • Impurities and pigments the addition of 6g/10ml of bentonite has the greatest adsorption capacity for protein impurities and pigments, and improves the purity of malt phytic acid.
  • Tea cake contains tea saponin, which has the functions of anti-inflammatory, analgesic and stomach-strengthening.
  • tea cake and rapeseed meal are good carbon sources and nitrogen sources for microbial fermentation.
  • the bentonite described in the step (1) has a particle size of 100 to 120 mesh.
  • the composite bacteria I in the step (2) is a combination of Bacillus natto with a seed age of 12 to 14 h and Lactobacillus rhamnosus with a seed age of 14 to 16 h.
  • the baicalin in the fermentation broth of Astragalus membranaceus can increase the phytic acid dissolution rate of malt; the natto bacteria in the fermentation period will produce nattokinase, superoxide dismutase and other active enzymes during the fermentation period, which can effectively hydrolyze glycosides.
  • the glucuronic acid bond in the element converts baicalin and baicalin in Astragalus membranaceus into baicalein and wogonin; the rhamnosyl lactic acid bacteria with the age of 14-16 h have the highest ability to ferment and release rhamnolipid, which can improve
  • the bacterium's vigor enhances the metabolism of natto bacteria, thereby promoting the conversion of baicalin and baicalin to baicalein and wogonin, reducing the overall fermentation time; Lactobacillus rhamnosus is a probiotic that can improve the response.
  • the activity of various enzymes in the system and the number of natto bacteria play a role in regulating the balance of the flora and improving the environment of the organism.
  • the composite bacteria II in the step (3) is a combination of Bacillus subtilis and Aspergillus niger having an age of 12 to 14 hours.
  • the mixed bacteria fermented tea cake and rapeseed meal is better than single-fermentation fermentation.
  • the fermentation of tea cake and rapeseed meal can be carried out by using Bacillus subtilis and Aspergillus niger into the logarithmic growth phase (age 12 ⁇ 14h). It is decomposed to produce various enzymes and tea saponin and phytic acid.
  • the chitosan has a degree of deacetylation of 50 to 60% as described in the step (4).
  • Chitosan has a good enzyme adsorption capacity, can adsorb the enzyme in the fermentation broth, and increase the amount of fermentation enzyme; the adsorption of chitosan can also clarify the fermentation broth and reduce the cost of downstream engineering; the degree of deacetylation is 50 ⁇ 60% of chitosan has the best water solubility and can be effectively dissolved in the fermentation broth;
  • the beneficial effects of the invention different enzymes and other by-products are produced by microbial fermentation, which promotes fermentation efficiency and shortens the fermentation cycle; the active ingredients of Chinese medicine and fruits and vegetables can completely retain their activity through microbial fermentation, and improve the white shrimp of South America
  • the stomach-enhancing effect of the feed additive the use of the oil by-product cake as a medium for the fermentation of tea saponin and phytic acid to promote the growth and efficiency of the fermentation bacterium, and improve the resource utilization rate; the enzyme liquid and vitamin prepared by the invention
  • the feed additive formulated with bran can effectively promote the food intake of P. vannamei, improve the immunity and disease resistance of the shrimp, and thereby increase the yield.
  • the temperature of the water immersed in the malt is 15-20 ° C.
  • the specific fermentation method is as follows: the mashed malt is added to the water of 1 time and 1% sodium chloride, and the 3% yeast is inoculated at 35 ° C for fermentation. 6h, a malt fermentation broth was obtained.
  • the fermentation broth of Astragalus membranaceus L was obtained, and then the composite bacterium I was inoculated into the fermentation broth of Astragalus membranaceus I, and fermented at 45 ° C, 100-200 rpm for 4 days to obtain the fermentation broth of Astragalus membranaceus;
  • the composite bacterium I is a combination of Bacillus natto liquid of 4% of 12h and 12% of Lactobacillus rhamnosus of Lactobacillus rhamnosus;
  • the composite bacteria II is a combination of Bacillus subtilis Bacillus subtilis with 4% of 12h and 12% of Aspergillus niger with 12h of 12h;
  • the temperature of the water immersed in the malt is 20 ° C.
  • the specific fermentation method is as follows: the mashed malt is added to the water of 1.5 times the mass and 1.2% of the sodium chloride, and the 3% yeast is inoculated at 35 ° C for 6 h. A malt fermentation broth is obtained.
  • the composite bacterium I is a combination of Bacillus natto bacillus of 6% of 14 h of age and 16 h of Lactobacillus rhamnosus of Lactobacillus rhamnosus;
  • the composite bacteria II is a combination of Bacillus subtilis broth of 6% of 14h and 14% of Aspergillus niger liquid of 14h;
  • the extract and the fermentation broth of Astragalus membranaceus were added, and the chitosan solution with a deacetylation degree of 60% of the fermentation broth III was added at the same time, and the fermentation was continued for 7 days after the fermentation was continued.
  • the filtrate is the enzyme solution; (5) 50 parts of the enzyme solution by weight ratio, added to 35 parts of bran, stirred and mixed, then added 1 part of vitamin C and 0.05 part of vitamin K solution, dried to The water content is 80%, and the feed additive of P. vannamei can be obtained.
  • the temperature of the water immersed in the malt is 15 ° C.
  • the specific fermentation method is as follows: the mashed malt is added to the water of 1.2 times the mass and the sodium chloride of 1.1%, and the 3% yeast is inoculated at 35 ° C for 6 h. A malt fermentation broth is obtained.
  • the composite bacterium I is a 5% natto Bacillus natto broth and a 4% larvae of Lactobacillus rhamnosus;
  • the composite bacteria II is a combination of Bacillus subtilis broth of 5% of 13h and 13h of Aspergillus niger;
  • the white shrimp feed used by the above users is the same product, and the feed additive is used as 10% of the total feed.
  • the color of the shrimp body is scored (5-point system). The higher the score, the better the body color.
  • the average score is as follows:

Abstract

一种南美白对虾饲料添加剂的制备方法，利用复合菌发酵茶饼粕、菜籽粕和黄芩，葡萄柚和麦芽分泌酵素，提高添加剂的营养成分，配以维生素和麸皮为载体制备而得。所制得的南美白对虾饲料添加剂可以显著减少南美白对虾养殖过程中病害的发生，提高虾的存活率，促进南美白对虾的生长，并能提高对虾肉品质和营养价值。其制备工艺简单，周期短，具有很好的发展前景。

Description

一种南美白对虾饲料添加剂的制备方法 技术领域
本发明属于水产养殖用饲料添加剂制备技术领域，特别涉及一种南美对虾饲料添加剂的制备方法。
背景技术
南美白对虾(Litopenaeusvannamei)属于对虾科，学名凡纳滨对虾，是世界养殖虾类产量最高的三大种类之一。南美白对虾个体大，壳薄体肥，出肉率高达65％以上，肉质鲜嫩，味道鲜美，蛋白质含量丰富，深受国内外市场欢迎，具有很高的经济价值；南美白对虾的人工养殖十分的广泛，是我国海水养殖的支柱产业。南美白对虾除了需要良好的生长环境、科学管理，还需要营养均衡的饲料，近年来由于养殖环境的恶化，养殖密度越来越高，导致南美白对虾易患病，生长缓慢；且由于现有的南美白对虾配合饲料营养部均衡、诱食性差等缺陷，且在水中稳定性不佳，也不利于南美白对虾进行快速有效的摄食，进一步加剧了生长缓慢、抗病力弱和成活率低。病害问题始终是阻碍南美白对虾养殖业可持续发展的瓶颈制约，特别是随着南美白对虾养殖的快速发展和集约化养殖程度的提高，以及采用人工饲料喂养，在养殖过程中经常出现大规模暴发性流行病和大批死亡的现象，而传统以抗生素和化学药物为主要手段的病害防治方法已经无法满足需求。其原因是，一方面大量使用抗生素和化学药物，导致病原体产生抗药性，防治效果下降，而防治成本显著提高；另一方面，由于药物残留的存在，直接影响消费群体的身体健康和公众安全。因此，现有的南美白对虾饲料难于满足南美对虾的使用要求，不利于南美白对虾饲养业的健康、可持续发展。
中草药具有残留少、致病菌不易产生抗药性等优点，其作用不仅针对致病微生物本身有直接的抑制和杀灭作用，更重要的是对饲喂对象免疫系统进行调整，提高饲喂动物的免疫机能和抗应激能力，将中草药成分制作成饲料添加剂，用于防治南美白对虾养殖中容易出现的病害完全符合发展无公害水产业、生产绿色水产品的病害防治准则。另外，中草药中汉有多种营养成分和生物活性物质，选择适当的中草药品种还可以起到促进南美白对虾生长和提高对虾品质的技术效果。
发明专利CN201310293138.0一种南美白对虾专用配合饲料，按重量百分比由以下组分组成：蒸汽鱼粉18-24％，豆粕17-22％，花生粕15-24％，血球蛋白粉2-4％，乌贼膏2～4％，酵母膏2-5％，面粉20-25％，鱼油2-3％，豆油1-2％，虾多维1.0％，虾多矿2.0％，南美白对虾专用饲料添加剂1.0％，以上各组分之和为100％。该发明的配合饲料的营养组成是根据南美白对虾的营养需求经反复的试验得出，其能有效地满足南美白对虾的营养需求，提高对虾的 免疫力和抗病能力，促进对虾的健康生长。
发明专利CN201210307270.8一种南美白对虾用诱食促长中草药复合饲料添加剂，由15～20％山楂，15～20％大黄，10～18％黄芪，10～14％甘草，10～15％阿魏，6～8％地龙，5～6％神曲，3～5％砂仁，3～5％干姜，4～5％牛磺酸，4～6％甜菜碱，牛磺酸和甜菜碱复合而成。该发明所述饲料添加剂可以对南美白对虾起到明显的诱食效果，可以取代供应日益紧张的动物性原料(尤其是鱿鱼膏)，以适应南美白对虾产业的发展需要。
发明专利CN201210307336.3一种预防南美白对虾肠炎病的中草药饲料添加剂，由龙胆、黄柏、陈皮、厚朴、大黄、茯苓、黄芩和黄芪，再加上玉米芯粉混合制成。该发明所述饲料添加剂可以用于预防南美白对虾肠炎病，具有提高南美白对虾的生长速度、增强对虾免疫力、提高养殖成功率的优点。
目前业界主要通过使用微生态制剂进行南美白对虾养殖水体的调节来进行南美白对虾肠炎病的预防，但因为目前微生态制剂的生产厂家众多产品质量良莠不齐，导致在实际养殖过程中发生肠炎病之后的预防效果不佳，给养殖户带来一定的养殖风险和成本浪费。
发明内容
针对现有技术中存在的上述不足，考虑到中草药具有对各种疾病优秀的治疗作用，因此可以在南美白对虾饲料中进行添加，从而达到减少南美白对虾养殖过程中存活率低的目的，本发明提供一种南美白对虾饲料添加剂的制备方法。
本发明是通过以下技术方实现的：
一种南美白对虾饲料添加剂的制备方法，由以下步骤制备而得：
(1)按重量份数比，取葡萄柚15～20份，剥皮分离果粒和果皮，然后将果粒进行榨汁，所得果汁中保留果肉，同时将果皮晒干并粉碎成粉末，得到葡萄柚果汁和葡萄柚皮粉末；取麦芽10～20份，加水侵泡20～24h后取出捣碎，加入酵母菌进行发酵，得到麦芽发酵液，然后将麦芽发酵液与葡萄柚果汁混合均匀，按总液质量的6～8％加入膨润土，用醋酸调节pH至1～2，室温下搅拌浸提1～3h后，离心去除膨润土沉淀，得到提取液；
(2)按重量份数比，取黄芩根15～20份，粉碎至粒度为40～50目后加入8～10倍量的蒸馏水，煎煮1.5～2h得到黄芩药液，待黄芩药液冷却至40～60℃后，于2000r/min下离心12～15分钟，取上清液，即得到黄芩提取液；按黄芩提取液的质量比加入1.0％蛋白胨，0.05％氯化钠，调节pH为7.0～8.6，高压灭菌并冷却至室温得到黄芩发酵液I，然后于黄芩发酵液I中接种复合菌I，在45～55℃、100～200r/min摇床上发酵4～6d，得到黄芩发酵液II；
按黄芩发酵液I体积比计，所述复合菌I为4～6％的纳豆菌Bacillus natto菌液和3～5％鼠 李糖乳杆菌Lactobacillus rhamnosus菌液组合；
(3)按重量份数比，取菜籽饼粕10～20份、茶饼粕10～20份，粉碎至50～100目后混合均匀，得到饼粕粉；将葡萄柚皮粉末、饼粕粉混合均匀，加入10～20倍量的水，混合均匀后高压灭菌，得到发酵液III，待其冷却后，接种复合菌II于35～40℃下进行发酵；
按发酵液III体积比计，所述复合菌II为4～6％的枯草芽孢杆菌Bacillus subtilis菌液和2～4％的黑曲霉Aspergillus niger菌液组合；
(4)发酵液III发酵6～7天后，加入提取液和黄芩发酵液II，同时加入发酵液III体积比5～10％的壳聚糖溶液，混合均匀继续发酵6～7天后，将发酵液过滤，滤液即为酵素液；
(5)按重量份数比，取酵素液45～50份，加入到35～45份麸皮中，搅拌混匀，然后加入0.1～1份维生素C和0.01～0.05份维生素K溶液，干燥至含水量为60～80％，即可得到南美白对虾饲料添加剂。
在步骤(1)中为了防止膨润土上吸附过多的提取物，可将第一次离心所得沉淀加入2倍量蒸馏水进行洗涤，再离心分离去除沉淀，将所得滤液与第一次离心分离所得滤液合并；
作为本发明的进一步改进，步骤(1)所述葡萄柚果皮晒干至含水量为10～20％，粉碎至15～20目。
粒度影响叶酸的浸出率，在15～20目叶酸能完整并最大化被提取浸出。
作为本发明的进一步改进，步骤(1)所述麦芽加水浸泡的水温为15～20℃，具体发酵方法如下：将捣碎后的麦芽加入其质量1～1.5倍的水和1～1.2％的氯化钠，接种3％的酵母菌于35℃下发酵6h，得到麦芽发酵液。
麦芽中的植酸随着麦芽的生长会逐渐减少，在低温下植酸活性降低，麦芽生长受到抑制，减少植酸的消耗；酵母发酵改变麦芽的结构和代谢，提高了植酸的产率。
葡萄柚中含有叶酸成分，促进肠胃氨基酸和糖代谢，提高胆碱合成，从而促进脂肪代谢，叶酸还能促进菜籽饼粕和油茶饼粕分解释放植酸和茶皂素；麦芽和菜籽粕中含有丰富的植酸，能促进菌体发酵效率，从而提高酵素含量；植酸水解后的磷能提高发酵产物酶的活力，从而提高酵素的营养价值；天然膨润土能有效吸附提取液中的蛋白质杂质和色素，投加量6g/10ml的膨润土对蛋白质杂质和色素的吸附能力最大，提高了麦芽植酸的纯度。
茶饼粕中含有茶皂素，具有消炎镇痛，健胃等功效，同时茶饼粕和菜籽粕是微生物发酵的良好碳源和氮源。
作为本发明的进一步改进，步骤(1)所述的膨润土粒度为100～120目。
作为本发明的进一步改进，步骤(2)所述复合菌I是由种龄为12～14h的纳豆菌Bacillus  natto和种龄为14～16h鼠李糖乳杆菌Lactobacillus rhamnosus组合。
黄芩发酵液中的黄芩素能提高麦芽植酸溶出率；种龄为12～14h纳豆菌在发酵过程中会生成纳豆激酶、超氧化物歧化酶等多种活性酶，能有效地水解苷元中的葡萄糖醛酸键，将黄芩中的黄芩苷和汉黄芩苷转化成黄芩素和汉黄芩素；种龄为14～16h的鼠李糖乳酸菌发酵分泌鼠李糖脂能力最大，能提高纳豆菌的菌活力，使纳豆菌的代谢增强，从而促进黄芩苷和汉黄芩苷转化成黄芩素和汉黄芩素，减少总体发酵时间；鼠李糖乳杆菌是一种益生菌，能提高反应体系中各种酶的活性和纳豆菌菌的数量，起到调节菌群的平衡，改善机体环境的作用。
作为本发明的进一步改进，步骤(3)所述复合菌II为种龄为12～14h的枯草芽孢杆菌Bacillus subtilis和的黑曲霉Aspergillus niger组合。
混合菌发酵茶饼粕和菜籽粕效果优于单菌发酵，利用进入对数生长期(种龄12～14h)的枯草芽孢杆菌和黑曲霉对茶饼粕和菜籽粕进行发酵，能最大化对其进行分解，产生各种酶和茶皂素与植酸。
作为本发明的进一步改进，步骤(4)所述的壳聚糖脱乙酰度为50～60％。
壳聚糖具有很好的酶吸附力，能将发酵液中的酶进行吸附，提高发酵酶量；壳聚糖的吸附作用还能使发酵液澄清，降低下游工程的成本；脱乙酰度50～60％的壳聚糖水溶性最好，能有效的溶解于发酵液中；
本发明的有益效果：通过微生物发酵产生不同的酶和其他副产物，促进了发酵效率，缩短了发酵周期；中药和果蔬的中有效成分通过微生物发酵能完整的保留其活性，提高了南美白对虾饲料添加剂的健胃作用效用；利用榨油副产物饼粕作为培养基发酵提取茶皂素和植酸促进体系发酵菌的生长和效率，提高了资源利用率；利用本发明制备的酵素液与维生素和麸皮复合配成的饲料添加剂能有效促进南美白对虾摄食量，提高虾体免疫力和抗病性，从而提高产量。
具体实施方式
下面结合实施例对本发明进一步说明。
实施例1
(1)按重量份数比，取葡萄柚15份，剥皮分离果粒和果皮，然后将果粒进行榨汁，所得果汁中保留果肉，同时将果皮晒干至含水量为10％，并粉碎成15目的粉末，得到葡萄柚果汁和葡萄柚皮粉末；取麦芽10份，加水侵泡20h后取出捣碎，加入酵母菌进行发酵，得到麦芽发酵液，然后将麦芽发酵液与葡萄柚果汁混合均匀，按总液质量的6％加入粒度为100目的膨润土，用醋酸调节pH至1～2，室温下搅拌浸提1h后，离心去除膨润土沉淀，得到提取液；
所述麦芽加水浸泡的水温为15～20℃，具体发酵方法如下：将捣碎后的麦芽加入其质量1倍的水和1％的氯化钠，接种3％的酵母菌于35℃下发酵6h，得到麦芽发酵液。
(2)按重量份数比，取黄芩根15份，粉碎至40目后加入8倍量的蒸馏水，煎煮1.5h得到黄芩药液，待黄芩药液冷却至40℃后，于2000r/min下离心12～15分钟，取上清液，即得到黄芩提取液；按黄芩提取液的质量比加入1.0％蛋白胨，0.05％氯化钠，调节pH为7.0～8.6，高压灭菌并冷却至室温得到黄芩发酵液I，然后于黄芩发酵液I中接种复合菌I，在45℃、100～200r/min摇床上发酵4d，得到黄芩发酵液II；
按黄芩发酵液I体积比计，所述复合菌I为4％种龄为12h的纳豆菌Bacillus natto菌液和3％种龄为14h鼠李糖乳杆菌Lactobacillus rhamnosus菌液组合；
(3)按重量份数比，取菜籽饼粕10份、茶饼粕10份，粉碎至50目后混合均匀，得到饼粕粉；将葡萄柚皮粉末、饼粕粉混合均匀，加入10倍量的水，混合均匀后高压灭菌，得到发酵液III，待其冷却后，接种复合菌II于35℃下进行发酵；
按发酵液III体积比计，所述复合菌II为4％种龄为12h的枯草芽孢杆菌Bacillus subtilis菌液和2～4％种龄为12h的黑曲霉Aspergillus niger菌液组合；
(4)发酵液III发酵6天后，加入提取液和黄芩发酵液II，同时加入发酵液III体积比5％脱乙酰度为50％的壳聚糖溶液，混合均匀继续发酵7天后，将发酵液过滤，滤液即为酵素液；
(5)按重量份数比，取酵素液45份，加入到45份麸皮中，搅拌混匀，然后加入0.1份维生素C和0.01份维生素K溶液，干燥至含水量为60％，即可得到南美白对虾饲料添加剂。
实施例2
(1)按重量份数比，取葡萄柚20份，剥皮分离果粒和果皮，然后将果粒进行榨汁，所得果汁中保留果肉，同时将果皮晒干至含水量为20％，并粉碎成20目的粉末，得到葡萄柚果汁和葡萄柚皮粉末；取麦芽20份，加水侵泡24h后取出捣碎，加入酵母菌进行发酵，得到麦芽发酵液，然后将麦芽发酵液与葡萄柚果汁混合均匀，按总液质量的8％加入粒度为120目的膨润土，用醋酸调节pH至1～2，室温下搅拌浸提3h后，离心去除膨润土沉淀，得到提取液；
所述麦芽加水浸泡的水温为20℃，具体发酵方法如下：将捣碎后的麦芽加入其质量1.5倍的水和1.2％的氯化钠，接种3％的酵母菌于35℃下发酵6h，得到麦芽发酵液。
(2)按重量份数比，取黄芩根20份，粉碎至50目后加入10倍量的蒸馏水，煎煮2h得到黄芩药液，待黄芩药液冷却至60℃后，于2000r/min下离心15分钟，取上清液，即得到黄芩提取液；按黄芩提取液的质量比加入1.0％蛋白胨，0.05％氯化钠，调节pH为8.6，高压灭菌并冷却至室温得到黄芩发酵液I，然后于黄芩发酵液I中接种复合菌I，在55℃、 100～200r/min摇床上发酵4～6d，得到黄芩发酵液II；
按黄芩发酵液I体积比计，所述复合菌I为6％种龄为14h的纳豆菌Bacillus natto菌液和5％种龄为16h鼠李糖乳杆菌Lactobacillus rhamnosus菌液组合；
(3)按重量份数比，取菜籽饼粕20份、茶饼粕20份，粉碎至100目后混合均匀，得到饼粕粉；将葡萄柚皮粉末、饼粕粉混合均匀，加入20倍量的水，混合均匀后高压灭菌，得到发酵液III，待其冷却后，接种复合菌II于40℃下进行发酵；
按发酵液III体积比计，所述复合菌II为6％种龄为14h的枯草芽孢杆菌Bacillus subtilis菌液和4％种龄为14h的黑曲霉Aspergillus niger菌液组合；
(4)发酵液III发酵7天后，加入提取液和黄芩发酵液II，同时加入发酵液III体积比10％脱乙酰度为60％的壳聚糖溶液，混合均匀继续发酵7天后，将发酵液过滤，滤液即为酵素液；(5)按重量份数比，取酵素液50份，加入到35份麸皮中，搅拌混匀，然后加入1份维生素C和0.05份维生素K溶液，干燥至含水量为80％，即可得到南美白对虾饲料添加剂。
实施例3
(1)按重量份数比，取葡萄柚18份，剥皮分离果粒和果皮，然后将果粒进行榨汁，所得果汁中保留果肉，同时将果皮晒干至含水量为15％，并粉碎成20目的粉末，得到葡萄柚果汁和葡萄柚皮粉末；取麦芽15份，加水侵泡20h后取出捣碎，加入酵母菌进行发酵，得到麦芽发酵液，然后将麦芽发酵液与葡萄柚果汁混合均匀，按总液质量的7％加入粒度为110目的膨润土，用醋酸调节pH至1～2，室温下搅拌浸提2h后，离心去除膨润土沉淀，得到提取液；
所述麦芽加水浸泡的水温为15℃，具体发酵方法如下：将捣碎后的麦芽加入其质量1.2倍的水和1.1％的氯化钠，接种3％的酵母菌于35℃下发酵6h，得到麦芽发酵液。
(2)按重量份数比，取黄芩根18份，粉碎至15目后加入9倍量的蒸馏水，煎煮2h得到黄芩药液，待黄芩药液冷却至50℃后，于2000r/min下离心13分钟，取上清液，即得到黄芩提取液；按黄芩提取液的质量比加入1.0％蛋白胨，0.05％氯化钠，调节pH为7.6，高压灭菌并冷却至室温得到黄芩发酵液I，然后于黄芩发酵液I中接种复合菌I，在50℃、100～200r/min摇床上发酵5d，得到黄芩发酵液II；
按黄芩发酵液I体积比计，所述复合菌I为5％种龄为13h的纳豆菌Bacillus natto菌液和4％种龄为15h鼠李糖乳杆菌Lactobacillus rhamnosus菌液组合；
(3)按重量份数比，取菜籽饼粕15份、茶饼粕16份，粉碎至80目后混合均匀，得到饼粕粉；将葡萄柚皮粉末、饼粕粉混合均匀，加入15倍量的水，混合均匀后高压灭菌，得到发酵液III，待其冷却后，接种复合菌II于37℃下进行发酵；
按发酵液III体积比计，所述复合菌II为5％种龄为13h的枯草芽孢杆菌Bacillus subtilis菌液和3％种龄为13h的黑曲霉Aspergillus niger菌液组合；
(4)发酵液III发酵6天后，加入提取液和黄芩发酵液II，同时加入发酵液III体积比7％脱乙酰度为55％的壳聚糖溶液，混合均匀继续发酵7天后，将发酵液过滤，滤液即为酵素液；
(5)按重量份数比，取酵素液48份，加入到42份麸皮中，搅拌混匀，然后加入0.8份维生素C和0.04份维生素K溶液，干燥至含水量为70％，即可得到南美白对虾饲料添加剂。
在广东江门地区，对20家农户的一个养殖塘进行试验，每5家为一组，使用的添加剂如下：
组1：使用实施例1制备的添加剂；
组2：使用实施例2制备的添加剂；
组3：使用实施例3制备的添加剂；
组4：不使用任何饲料添加剂；
以上用户使用的对白虾饲料均为同种产品，饲料添加剂使用量为投食饲料总量的10％。
从虾种投放到收获，记录收获后的白对虾养殖效果，具体如表1。
表1 南美白对虾养殖效果
组别 发病率(％) 成活率(％) 养殖周期(天) 10只虾总重量(g)
1 6.8 92.8 82 365
2 6.1 92.1 84 371
3 5.6 93.5 81 382
4 18.2 81.1 102 242
同时对虾体颜色进行打分(5分制)，分值越高体色越好，平均得分如下：
组1：4.8分；
组2：4.8分；
组3：4.9分；
组4：4.1分。
通过以上试验可知，本发明制备的南美白对虾饲料添加剂具有很好的效益。
以上所述，仅为利用本创作技术内容的实施例，任何熟悉本项技术者运用本创作所做的修饰、变化，皆属本创作主张的专利范围。

Claims (7)

  1. 一种南美白对虾饲料添加剂的制备方法，其特征在于，包括以下步骤：
    (1)按重量份数比，取葡萄柚15～20份，剥皮分离果粒和果皮，然后将果粒进行榨汁，所得果汁中保留果肉，同时将果皮晒干并粉碎成粉末，得到葡萄柚果汁和葡萄柚皮粉末；取麦芽10～20份，加水侵泡20～24h后取出捣碎，加入酵母菌进行发酵，得到麦芽发酵液，然后将麦芽发酵液与葡萄柚果汁混合均匀，按总液质量的6～8％加入膨润土，用醋酸调节pH至1～2，室温下搅拌浸提1～3h后，离心去除膨润土沉淀，得到提取液；
    (2)按重量份数比，取黄芩根15～20份，粉碎至粒度为40～50目后加入8～10倍量的蒸馏水，煎煮1.5～2h得到黄芩药液，待黄芩药液冷却至40～60℃后，于2000r/min下离心12～15分钟，取上清液，即得到黄芩提取液；按黄芩提取液的质量比加入1.0％蛋白胨，0.05％氯化钠，调节pH为7.0～8.6，高压灭菌并冷却至室温得到黄芩发酵液I，然后于黄芩发酵液I中接种复合菌I，在45～55℃、100～200r/min摇床上发酵4～6d，得到黄芩发酵液II；
    按黄芩发酵液I体积比计，所述复合菌I为4～6％的纳豆菌Bacillus natto菌液和3～5％鼠李糖乳杆菌Lactobacillus rhamnosus菌液组合；
    (3)按重量份数比，取菜籽饼粕10～20份、茶饼粕10～20份，粉碎至50～100目后混合均匀，得到饼粕粉；将葡萄柚皮粉末、饼粕粉混合均匀，加入10～20倍量的水，混合均匀后高压灭菌，得到发酵液III，待其冷却后，接种复合菌II于35～40℃下进行发酵；
    按发酵液III体积比计，所述复合菌II为4～6％的枯草芽孢杆菌Bacillus subtilis菌液和2～4％的黑曲霉Aspergillus niger菌液组合；
    (4)发酵液III发酵6～7天后，加入提取液和黄芩发酵液II，同时加入发酵液III体积比5～10％的壳聚糖溶液，混合均匀继续发酵6～7天后，将发酵液过滤，滤液即为酵素液；
    (5)按重量份数比，取酵素液45～50份，加入到35～45份麸皮中，搅拌混匀，然后加入0.1～1份维生素C和0.01～0.05份维生素K溶液，干燥至含水量为60～80％，即可得到南美白对虾饲料添加剂。
  2. 根据权利要求1所述的一种南美白对虾饲料添加剂的制备方法，其特征在于，步骤(1)所述葡萄柚果皮晒干至含水量为10～20％，粉碎至15～20目。
  3. 根据权利要求1所述的一种南美白对虾饲料添加剂的制备方法，其特征在于，步骤(1)所述麦芽加水浸泡的水温为15～20℃，具体发酵方法如下：将捣碎后的麦芽加入其质量1～1.5倍的水和1～1.2％的氯化钠，接种3％的酵母菌于35℃下发酵6h，得到麦芽发酵液。
  4. 根据权利要求1所述的一种南美白对虾饲料添加剂的制备方法，其特征在于，步骤(1)所述的膨润土粒度为100～120目。
  5. 根据权利要求1所述的一种南美白对虾饲料添加剂的制备方法，其特征在于，步骤(2) 所述复合菌I是由种龄为12～14h的纳豆菌Bacillus natto和种龄为14～16h鼠李糖乳杆菌Lactobacillus rhamnosus组合。
  6. 根据权利要求1所述的一种南美白对虾饲料添加剂的制备方法，其特征在于，步骤(3)所述复合菌II为种龄为12～14h的枯草芽孢杆菌Bacillus subtilis和的黑曲霉Aspergillus niger组合。
  7. 根据权利要求1所述的一种南美白对虾饲料添加剂的制备方法，其特征在于，步骤(4)所述的壳聚糖脱乙酰度为50～60％。
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