CN104938787A - Cottonseed meal compound microbial ecological agent suitable for aquaculture and preparation and application - Google Patents

Cottonseed meal compound microbial ecological agent suitable for aquaculture and preparation and application Download PDF

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Publication number
CN104938787A
CN104938787A CN201510389217.0A CN201510389217A CN104938787A CN 104938787 A CN104938787 A CN 104938787A CN 201510389217 A CN201510389217 A CN 201510389217A CN 104938787 A CN104938787 A CN 104938787A
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dregs
cotton dregs
pulvis
agent
preparation
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CN104938787B (en
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王亮
闫卫疆
高宏伟
孙雪梅
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XINJIANG TAIKUN Group LLC
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XINJIANG TAIKUN GROUP CO Ltd
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    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81Aquaculture, e.g. of fish
    • Y02A40/818Alternative feeds for fish, e.g. in aquacultures
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87Re-use of by-products of food processing for fodder production

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  • Feed For Specific Animals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Fodder In General (AREA)

Abstract

The invention discloses a cottonseed meal compound microbial ecological agent suitable for aquaculture and preparation and application. The cottonseed meal compound set agent comprises 60%-70% of cottonseed meal, 8%-10% of sunflower seed meal, 15%-13% of rapeseed meal, 5%-12% of safflower seed meal, 2%-5% of grape pip meal and 1.5% of added multiple species inoculants; by using solid state fermentation, the cottonseed meal compound set agent is prepared, the multiple species inoculants are prepared and obtained by mixing of bacillus coagulans inoculants, candida utilis inoculants and Aspergillus niger inoculants in a proportion of 3:5:2, a direct mixed bacteria powder agent is composed of a bacillus cereus powder agent and a lactobacillus plantarum powder agent in a proportion of 1:1, and according to the fact that every 500 g of the direct mixed bacteria powder agent and 500 kg of the cottonseed meal compound set agent are mixed, the compound microbial ecological agent is prepared. The prepared cottonseed meal compound microbial ecological agent is suitable for the aquaculture such as fish culture, the true nutritive value of fishes is high, the water stability is good, the water body pollution is small, and technical effect of promoting fish body growing and other obvious technical effects are achieved.

Description

A kind of cotton dregs compound micro-ecological preparation and Synthesis and applications being suitable for aquaculture
Technical field
The invention belongs to the technical field of modern feed industry.Specifically, the present invention relates to a kind of technical field being applicable to the cotton dregs compound micro-ecological preparation preparations and applicatio of aquaculture.
Background technology
Fast, increasing biotechnology product also developed by the core technology feed addictive for feed industry to China's feedstuff industry development along with the development of biotechnology.Microbial forage additive arises at the historic moment, the application study of probiotics overcomes all drawbacks that Long-Time Service antibiotic causes to aquaculture, have broad application prospects to development green feed additive, oneself is classified as 21st century emphasis disseminate technology project by the Ministry of Agriculture.This means the research to probiotics, popularization, meet the general requirement of China's sustainable development.A lot of pharmacy corporation all adds the research and development of probiotics and produces ranks at present, and professional standard is specification gradually.Scientific research is complete gradually to the industrial chain of producing marketing.
The U.S. brings into use feeding micro-ecological preparation from the seventies, and Japan, some developed countries of West Europe develop novel Substitutes For Antibiotic in succession, probiotics is obtained universal rapidly.Along with the development of science and technology, feeding micro-ecological preparation has improvement greatly in bacterial classification, production technology, formulation, and stability, the resist processing of product also improve greatly.
The research of China's feeding micro-ecological preparation starts from the eighties, in succession has the nineties product to occur, 2010 registered has feeding micro-ecological preparation hundreds of.Current China market supplying feeding micro-ecological preparation various in style, comprise single microbial inoculum, is more composite bacteria agent capable, and registered external imported product has tens kinds, and constantly has new product introduction.It is reported, domestic microbiological feed addictive industry is in the starting stage at present, and the probiotics market space is very big, and prospect of industrial development is very wide.
China is the country that a protein resource is rare.Within 2011, the Chinese dependency degree to imported soybean is more than 80%; Latter stage in 20th century, the annual approximately import 700,000 tons of fish meal of China, about 80% from Peru, from Chilean import volume less than 10%, also has a small amount of import in addition from the U.S., Japan, country in Southeast Asia; Internal feed industry is day by day urgent to the demand of protein raw materials of high quality and at a reasonable price.According to the estimation of national feed office, according to the development plan requirement of our people's diet structure and aquaculture, have larger breach to the protein feed needed for the year two thousand twenty.In order to solve the high and problem of inadequate resource of current common protein feed price, people start the assorted dregs of rice feeds such as development and utilization cotton dregs, sunflower meal.Although the amino acid digestibility in these protein feeds is low, limiting amino acid content is low, harmony right between amino acid is poor, and containing certain anti-nutrient substance, if but through suitable processing process and Appropriate application, still can part even replacing whole common protein feed, increase economic efficiency.
Cottonseed is the edible oil and fat resource with soybean, dish dregs of rice no less important, is that one develops protein resource completely.Xinjiang is the maximum cotton in the whole nation and oily certain herbaceous plants with big flowers growing area, have high-quality, abundant oil plant and protein raw materials resource, grease processing industry in Xinjiang only focuses on the quality of edible oil for a long time, have ignored the exploitation of dregs of rice series products, the low albumen dregs of rice class after oil expression can only use than row as low side raw material is low in feed.
The exploitation of probiotics and extensive use, to promoting that the development of China's aquaculture has played positive effect, and potentiality are very large.At present, continue photosynthetic bacteria in aquatic products after extensive use, many probioticses progressively penetrate into aquatic products industry, and play its significant advantage, and along with the development of " white agriculture ", probiotics will play an important role in aquaculture.At present, about the application of probiotics in aquaculture, Chinese scholars has been carried out some and has been attempted, and obtains ideal effect, but than the investigation and application in medical science and herding, still has larger gap, be still in reduced levels.In view of the particular surroundings of aquaculture, and developing in depth and breadth along with bioscience, after the antibiotic epoch of radiance, will be new probiotics epoch, its application in aquaculture will have prospect more widely.The exploitation of green feed additive has been trend of the times, it is worthy of note, while research probio, must under zoologize the feature of enteron aisle bacterium flora self and the relation between host and environment energetically, just can design efficient probiotics of new generation.
Current probiotics is transporting, is using and easy in inactivation in preservation process, thus reduces its biological action; After some bacterial strain enters alimentary canal, be not highly resistant to the effect of hydrochloric acid, bile acid etc., thus function cannot be played by the enteron aisle of field planting for a long time; Probiotics preventive effect is better than result for the treatment of, but effect is slow, needs long-time continuous to use, unfavorable factor and the limitation of the technology such as the corresponding increase of expense.Above problem needs to further investigate Application and Development further.
Summary of the invention
Employing cotton dregs are had no for prior art, sunflower seed dregs, the dish dregs of rice, safflower seed meal and grape pip dregs of rice mixture are fermentation substrate, utilize fermentable to combine with direct putting type probiotics and prepare the novel state of the art being applicable to the cotton dregs compound micro-ecological preparation of aquaculture, the present invention is intended to provide a kind of the cotton dregs compound micro-ecological preparation and the Synthesis and applications that are suitable for aquaculture, decomposed by fermentable and improve cotton dregs, sunflower seed dregs, the dish dregs of rice, the quality of safflower seed meal and the grape pip dregs of rice, and prepare the agent of cotton dregs complex group, the technology of preparing of probiotics is utilized to prepare the direct putting type microorganism lyophilized formulations being suitable for it and coordinating, by cotton dregs complex group agent standby for fermentation to be coordinated aquacultures such as being suitable for fish with direct putting type microorganism lyophilized formulations compound, guarantee that the digestibility of fish body is high, waste is few, water stability is good, little to water pollution, the sour-sweet fragrance of smell, good palatability and the appetite of fish can be promoted, promote fish bulk-growth.
Technical scheme provided by the invention:
Cotton dregs compound micro-ecological preparation of the present invention is by the agent of cotton dregs complex group and deliver directly mixed bacteria pulvis and form, and the agent of cotton dregs complex group contains cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and the grape pip dregs of rice, is prepared from by solid fermentation; Wherein primary raw material selects cotton dregs 60%-70%, sunflower seed dregs 8%-10%, dish dregs of rice 15%-13%, safflower seed meal 5%-12% and grape pip dregs of rice 2%-5%, mix bacterium agent is made up of bacillus coagulans, candida utili and aspergillus niger microbial inoculum, prepares the agent of cotton dregs complex group by solid state fermentation.Adopt common preparation method to obtain bacillus cereus pulvis and Lactobacillus plantarum pulvis respectively in conjunction with freeze drying technology preparation to form and deliver directly mixed bacteria pulvis.By the agent of cotton dregs complex group with deliver directly mixed bacteria pulvis mixing composition cotton dregs compound micro-ecological preparation, be applied to the aquacultures such as fish, for effectively utilizing cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and grape pip dregs of rice resource, fully meet the demand of the aquacultures such as fish, for guaranteeing that the digestibility of fish body is high, waste less, water stability good, little to water pollution, the sour-sweet fragrance of smell, good palatability and can promote fish appetite, promote that the aspects such as fish bulk-growth obtain significantly outstanding technique effect.
The present invention specifically provides a kind of cotton dregs compound micro-ecological preparation being suitable for aquaculture, by the agent of cotton dregs complex group with deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 60%-70%, sunflower seed dregs 8%-10%, dish dregs of rice 15%-13%, the composite bacteria agent capable of safflower seed meal 5%-12% and grape pip dregs of rice 2%-5% and 1.5% prepares the agent of cotton dregs complex group by solid state fermentation, and composite bacteria agent capable mixes acquisition by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum by weight 3:5:2 ratio; Deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix; Be mixed with according to the cotton dregs complex group agent delivering directly mixed bacteria pulvis and 500 kilograms of every 500g and obtain cotton dregs compound micro-ecological preparation.
Meanwhile, the present invention specifically provides a kind of preparation method being suitable for the cotton dregs compound micro-ecological preparation of aquaculture, prepares especially by following technique, and % is according to weight percent meter, and concrete preparation method's step is as follows:
(1) pretreatment of raw material: by raw material cotton dregs 60%-70%, sunflower seed dregs 8%-10%, dish dregs of rice 15%-13%, safflower seed meal 5%-12% and grape pip dregs of rice 2%-5%, mix in proportion and pulverize, and crosses 40 mesh sieves; Add water according to the solid-liquid ratio of weight ratio 1:0.8, be uniformly mixed acquisition raw material.
(2) acquisition of the activation of bacterial classification, cultivation and microbial inoculum: respectively bacillus coagulans, candida utili and the aspergillus niger selected are carried out test tube slant activation culture, and the bacterial classification activated through test tube slant is transferred in shaking flask cultivates, obtain bacillus coagulans microbial inoculum, candida utili microbial inoculum and black-koji mould microbial inoculum respectively.
(3) preparation of composite bacteria agent capable: bacillus coagulans microbial inoculum, candida utili microbial inoculum and the aspergillus niger microbial inoculum ratio according to weight ratio 3:5:2 is mixed, namely prepares composite bacteria agent capable.
(4) bacterium is connect: the composite bacteria agent capable prepared by step (3) is linked in raw material prepared by step (1) by the inoculative proportion with 1.5%.
(5) solid fermentation: adopt common Zymolysis Equipment, raw material prepared by whipping step (1) and the composite bacteria agent capable that step (3) is added, make it to mix, and is 30 DEG C-35 DEG C through controlling fermentation temperature, pH nature, continuous solid-state fermentation 48h-60h.
(6) dry, granulate: the raw material that above-mentioned steps (5) fermentation is got ready through granulation, dry to moisture be 12% acquisition cotton dregs complex group agent.
(7) preparation of bacterial classification pulvis: select bacillus cereus and Lactobacillus plantarum to obtain pulvis through the activation of bacterial classification, the collection of thalline, the freeze drying of bacterium mud respectively:
The activation of bacterial classification: the bacillus cereus selected and Lactobacillus plantarum are carried out first time by the MRS liquid culture medium of 100ml sterilizing respectively and activates, activation temperature 37 DEG C, 24h rear plate detects purity and viable count, detect up to standard after more above-mentioned 100ml bacterium liquid is all proceeded to 400ml sterilizing MRS liquid culture medium carry out second time activation, 24h rear plate detects purity and viable count, detect up to standard after go in fermentation tank and carry out expansions cultivation 48h.
Microorganism collection: utilize the bacterium liquid of ultracentrifuge to the bacillus cereus after fermentation and Lactobacillus plantarum to carry out centrifugal respectively, be centrifugal 10min under the condition of 4000-6000r/min at rotating speed, go supernatant to collect deposit.
Bacterium mud freeze drying: the deposit of the bacillus cereus of collecting respectively and Lactobacillus plantarum is added to composite protective agent, with oscillator mixing, makes bacteria suspension; Contained in freezing plate again, after first putting 4 DEG C of refrigerator balance 30min subsequently, move into-30 DEG C of refrigerator 60min, then-80 DEG C of refrigerator 60min are put, freeze drying 14h on frozen vacuum dryer again, sealing is stored in 4 DEG C of refrigerators, prepares bacillus cereus pulvis and Lactobacillus plantarum pulvis respectively.Experimentally design, vacuum freeze drying condition setting freeze-drying instrument cavity temperature is-60 DEG C-70 DEG C, and pressure is 0.006Pa, and the time is 14h; Composite protective agent adopts skimmed milk power 7.06%, maltose 6.46% and sodium glutamate 6.7%.
(8) allocate: bacillus cereus pulvis above-mentioned steps obtained respectively and Lactobacillus plantarum pulvis form acquisition according to weight ratio 1:1 and deliver directly mixed bacteria pulvis.
(9) finished product: the cotton dregs complex group agent of selecting above-mentioned steps (6) to prepare and step (8) prepare delivers directly mixed bacteria pulvis, to mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
In the present invention, selected bacillus coagulans, candida utili, aspergillus niger, bacillus cereus and Lactobacillus plantarum are all common bacterial classification, and those of ordinary skill in the art can be obtained by public's channel.
Bacillus coagulans selected by the present invention, candida utili, aspergillus niger, although bacillus cereus and Lactobacillus plantarum are all common bacterial classification, but, the specificity of microorganism fungus kind and complexity, various different bacterial classification can be used by compound compatibility, by the blending of various bacterial classification, compatibility is combined with each bacterial classification attribute, consider the security of composite bacteria, particularly apply in the aquacultures such as fish, all need a large amount of infrastest checkings, the present invention is based on basic research accumulation in early stage, by adopting the composite test of bacterial classifications different in a large number, prove that the present invention adopts composite bacteria agent capable to prepare the agent of cotton dregs complex group by solid state fermentation, composite bacteria agent capable is by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum obtain according to the ratio mixing of weight ratio 3:5:2, for the selection delivering directly mixed bacteria, take into full account the attribute of bacterial classification and deliver directly mixed bacteria for employing cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and grape pip dregs of rice compound carry out comprehensive consideration by the applicability bringing aquaculture application after solid state fermentation, based on a large amount of scientific experiment, determine that selecting bacillus cereus and Lactobacillus plantarum to adopt prepares freeze dried powder respectively, with prepared the composite blending of composite bacteria agent capable by solid state fermentation, thus entirety prepares the cotton dregs complex group agent having and be suitable for fish aquaculture, visible, will by the composite and practicality of bacterial classification, cotton dregs complex group agent component setting provided by the invention and component are really established a capital and are obtained through a large amount of scientific experiment, and by application attestation for how to fully utilize cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and the grape pip dregs of rice, decompose the wherein poisonous and harmful substance such as such as gossypol, for aquacultures such as application fish, guarantee that the digestibility of fish body is high, waste is few, water stability is good, little to water pollution, the sour-sweet fragrance of smell, good palatability and the appetite of fish can be promoted, promote that the aspects such as fish bulk-growth obtain significant technique effect.
Throw type leaven is a kind of novel fermentation agent not needing activation and amplification just can directly apply to production, there is the advantages such as viable bacteria content is high, bacterial classification long shelf-life, also prevent spawn degeneration and pollution, improve labor productivity and the product quality of fermented product industry.Because bacterial classification is to the sensitiveness of thermostimulation, throw type leaven technique adopts vacuum freeze-drying method more, in freezing and dry run, part cellular damage, death and some protease molecule passivation can be made.In order to improve the survival rate after bacterial classification freeze-drying, it is necessary for adding protective agent minimizing Freeze-drying Damage before freeze-drying, makes it play defencive function to greatest extent, improves the survival rate of fermenting agent.Different protective agents respectively has pluses and minuses; single protective agent often can not meet the condition that thalline resists extraneous adverse circumstances; in composite protective agent, each protective agent then plays respective characteristic and can play synergistic effect in freeze drying; the present invention obtains bacillus cereus by verification experimental verification and the protectant optimum formula of Lactobacillus plantarum is skimmed milk power 7.06%, maltose 6.46% and sodium glutamate 6.7%; Lactobacillus plantarum freeze drying survival rate is to 92.95%, and the freeze drying survival rate of bacillus cereus is to 94.1%.
This area bacillus cereus and the common screening MRS culture medium of Lactobacillus plantarum: peptone 10g, yeast extract 5g, beef extract 10g, glucose 20g, dipotassium hydrogen phosphate 2g, citric acid tri-amonia 1g, sodium acetate 5g, magnesium sulfate 0.8g, manganese sulfate 0.25g, Tween 80 1ml, pH value 6.2-6.4, be settled to 1000ml with distilled water, 115-120 DEG C of sterilizing 30min.
This area bacillus cereus and Lactobacillus plantarum isolation medium are selected and common are separated MRS culture medium: tryptone 10g, beef extract 10g, dibasic ammonium citrate 2g, sodium acetate 5g, dusty yeast 5g, glucose 5g, potassium dihydrogen phosphate 2g, Tween 80 1.0ml, calcium carbonate 20g, magnesium sulfate 0.58g, manganese sulfate 0.25g, water 1000ml, adjust pH 6.2-6.5,115 DEG C of sterilizing 20min.
The modified MRS culture medium that bacillus cereus of the present invention and Lactobacillus plantarum fermented and cultured adopt: add α-methylglucoside 10g, potassium sorbate 1g on MRS medium base, adjust pH 6.2-6.5,115 DEG C of sterilizing 20min; Culture medium prescription after change is, dusty yeast 3g/L, peptone 5g/L, beef extract 2g/L, MnSO 40.005g/L, NaCl 2g/L, K 2hPO 43g/L, MgSO 40.02g/L, this medium base carries out the optimization of fermentation condition, show that the condition of culture after optimization is: temperature 40 DEG C, initial pH value is 7.0, rotating speed 210r/min, and liquid amount is 30mL (triangular flask of 250mL), inoculum concentration is 6% (v/v), fermentation time 48h.
Cotton dregs compound micro-ecological preparation good palatability prepared by the technology of the present invention, can promote the appetite of fish; There is good formability and resistance to water, ensure that the stability of cotton dregs compound micro-ecological preparation in water, reduce the pollution of water body; Meanwhile, the digestibility in fish body is high, promotes the growth of fish body, is with a wide range of applications.
The cotton dregs that the present invention adopts, sunflower seed dregs, the dish dregs of rice, the pulverizing of safflower seed meal and the grape pip dregs of rice, add water, the pretreatment such as mixing, the activation of bacterial classification, fermentation, although the technological means such as the freeze drying of bacterial classification be unable to do without support and the inspiration of prior art, but the material ratio of cotton dregs compound micro-ecological preparation in zymotechnique, solid-liquid ratio, the selection of fermented bacterium and deliver directly the selection of mixed bacteria, preparation, for determining that the quality of cotton dregs compound micro-ecological preparation all has important effect, also be deciding factor, technological means and the instrument of not talkative employing are prior aries, and then negate the novelty that entirety provided by the invention prepares cotton dregs compound micro-ecological preparation, existing all technological inventions and innovation all be unable to do without support and the basis of prior art.Visible, each technical factor in cotton dregs compound micro-ecological preparation preparation technology provided by the invention and technical parameter are all through comprehensive consideration and design, in the preparation method of cotton dregs compound micro-ecological preparation provided by the invention, each sport technique segment has one up and down, any technical step is indispensable, tight indivisible feature, can not approbate and reprobate dismemberent technical step wherein.
Further, the invention provides and utilize the above-mentioned application of cotton dregs compound micro-ecological preparation in aquaculture prepared.Good palatability, can promote the appetite of fish; There is good formability and resistance to water, ensure that the stability of cotton dregs mixed feed in water, reduce the pollution of water body; Meanwhile, the digestibility in fish body is high, is with a wide range of applications.
Following beneficial effect can be reached by the summary of the invention implementing the present invention concrete:
(1) the present invention is by utilizing cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and grape pip dregs of rice mixture to be raw material, microbial fermentation technology is utilized to prepare the agent of cotton dregs complex group, prepare direct putting type probiotics by freeze drying technology, match by utilizing the agent of cotton dregs complex group and direct putting type probiotics and prepare the novel cotton dregs compound micro-ecological preparation being applicable to aquaculture.The technical issues that need to address of the present invention be just to overcome prior art defect, make full use of the protein raw materials such as cotton dregs, sunflower seed dregs, the sour-sweet fragrance of cotton dregs compound micro-ecological preparation smell of preparation, good palatability, can promote the appetite of fish; There is good formability and resistance to water, ensure that the stability of cotton dregs mixed feed in water, reduce the pollution of water body; Meanwhile, the digestibility in fish body is high, is with a wide range of applications.
(2) the cotton dregs compound micro-ecological preparation being suitable for aquaculture provided by the invention, by taking into full account the nutritional characteristic of cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and the grape pip dregs of rice, cotton dregs mixed feed is prepared through solid-fermented technique, prepare direct putting type probiotics through freeze drying technology, match by utilizing the agent of cotton dregs complex group and direct putting type probiotics and prepare the novel cotton dregs compound micro-ecological preparation being applicable to aquaculture.The cotton dregs compound micro-ecological preparation of preparation contains the nutriments such as rich in protein, vitamin, mineral matter, is of high nutritive value, can promotes the growth of fish body.
(3) adopt the cotton dregs compound micro-ecological preparation cost being suitable for aquaculture provided by the invention low, the output of the cotton dregs such as cotton dregs, sunflower seed dregs compound micro-ecological preparation raw material is large, cost recovery is low, and equipment is simple in the process preparing cotton dregs compound micro-ecological preparation, technique is simple, and the cost of preparation is low.
(4) adopt the cotton dregs compound micro-ecological preparation being suitable for aquaculture provided by the invention good at water stability: cotton dregs compound micro-ecological preparation is statically placed in water body, all float on water surface, after soaking 15min, feed granules starts to sink, final nearly half feed is in water body upper strata, illustrate that cotton dregs compound micro-ecological preparation has good water stability, little to water pollution.
(5) the cotton dregs compound micro-ecological preparation being suitable for aquaculture provided by the invention is adopted can to promote fish bulk-growth: cotton dregs compound micro-ecological preparation demonstrates good effect in carp growth, wherein the quality of carp unit length fish body is large, fish body grossness, fish body trunk ratio is large, edible part is many, promotes the growth of fish body.
Accompanying drawing explanation
Fig. 1 is shown as the process flow diagram of cotton dregs compound micro-ecological preparation.
Detailed description of the invention
, for embodiment, the present invention is described below, but the present invention is not limited to following embodiment.
The primary raw material selected in the present invention: cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and grape pip dregs of rice raw material are all well known selecting; Selected bacillus coagulans, candida utili, aspergillus niger, bacillus cereus and Lactobacillus plantarum are all common bacterial classification, and those of ordinary skill in the art can be obtained by public's channel.
The capital equipment selected: the equipment such as pulverizer, agitator, high-pressure sterilizing pot, incubator, aseptic operating platform, temperature control shaking table, supercentrifuge, fermentation tank, vacuum refrigeration centrifuge, bacterium liquid bottle are all well known selecting.
The all bacterial classifications selected in the present invention and raw and auxiliary material, and the Spawn incubation condition selected and method are all well known selecting, the % related in the present invention is weight percentage, unless otherwise indicated.
But do not limit enforcement of the present invention, in addition, in following explanation, if no special instructions, all % related to all refer to m/m mass percent or weight part ratio meter.
Embodiment one: the cotton dregs compound micro-ecological preparation being suitable for aquaculture
The present invention specifically provides a kind of cotton dregs compound micro-ecological preparation being suitable for aquaculture, by the agent of cotton dregs complex group with deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 60%-70%, sunflower seed dregs 8%-10%, dish dregs of rice 15%-13%, the composite bacteria agent capable of safflower seed meal 5%-12% and grape pip dregs of rice 2%-5% and 1.5% prepares the agent of cotton dregs complex group by solid state fermentation, and composite bacteria agent capable mixes acquisition by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum according to the ratio of weight ratio 3:5:2; Deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix; To mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
In the present invention, selected bacillus coagulans, candida utili, aspergillus niger, bacillus cereus and Lactobacillus plantarum are all common bacterial classification, those of ordinary skill in the art can be obtained by public's channel, meanwhile, the preparation method of above-mentioned strain fermentating liquid that adopts of the present invention and the culture medium selected in preparation process thereof and condition of culture be all this area common with know.
Bacillus coagulans selected by the present invention, candida utili, aspergillus niger, although bacillus cereus and Lactobacillus plantarum are all common bacterial classification, but, the specificity of microorganism fungus kind and complexity, various different bacterial classification can be used by compound compatibility, by the blending of various bacterial classification, compatibility is combined with each bacterial classification attribute, consider the security of composite bacteria, particularly apply in the aquacultures such as fish and all need a large amount of infrastest checkings, the present invention is based on basic research accumulation in early stage, by adopting the composite test of bacterial classifications different in a large number, prove that the present invention adopts composite bacteria agent capable to prepare the agent of cotton dregs complex group by solid state fermentation, composite bacteria agent capable is by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum obtain according to the ratio mixing of weight ratio 3:5:2, for the selection delivering directly mixed bacteria, take into full account the attribute of bacterial classification and deliver directly mixed bacteria for employing cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and grape pip dregs of rice compound are by bringing the comprehensive consideration of the applicability of the application of aquaculture after solid state fermentation, based on a large amount of scientific experiment, determine that selecting bacillus cereus and Lactobacillus plantarum to adopt prepares freeze dried powder respectively, the composite blending of composite bacteria agent capable is prepared by solid state fermentation, thus entirety prepares the cotton dregs complex group agent had for being suitable for fish aquaculture, visible, will by the composite and practicality of bacterial classification, cotton dregs complex group agent component setting provided by the invention and component are really established a capital and are obtained through a large amount of scientific experiment, and by application attestation for how to fully utilize cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and the grape pip dregs of rice, decompose the wherein poisonous and harmful substance such as such as gossypol, for aquacultures such as application fish, guarantee that the digestibility of fish body is high, waste is few, water stability is good, little to water pollution, the sour-sweet fragrance of smell, good palatability and the appetite of fish can be promoted, promote that the aspects such as fish bulk-growth obtain significant technique effect.
Throw type leaven is a kind of novel fermentation agent not needing activation and amplification just can directly apply to production, there is the advantages such as viable bacteria content is high, bacterial classification long shelf-life, also prevent spawn degeneration and pollution, improve labor productivity and the product quality of fermented product industry.Because bacterial classification is to the sensitiveness of thermostimulation, throw type leaven technique adopts vacuum freeze-drying method more, in freezing and dry run, part cellular damage, death and some protease molecule passivation can be made.In order to improve the survival rate after bacterial classification freeze-drying, it is necessary for adding protective agent minimizing Freeze-drying Damage before freeze-drying, makes it play defencive function to greatest extent, improves the survival rate of fermenting agent.Different protective agents respectively has pluses and minuses; single protective agent often can not meet the condition that thalline resists extraneous adverse circumstances; in composite protective agent, each protective agent then plays respective characteristic and can play synergistic effect in freeze drying; the present invention obtains bacillus cereus by verification experimental verification and the protectant optimum formula of Lactobacillus plantarum is skimmed milk power 7.06%, maltose 6.46% and sodium glutamate 6.70%; Lactobacillus plantarum freeze drying survival rate is to 92.95%, and the freeze drying survival rate of bacillus cereus is to 94.1%.
This area bacillus cereus and the common MRS culture medium of Lactobacillus plantarum: peptone 10g, yeast extract 5g, beef extract 10g, glucose 20g, dipotassium hydrogen phosphate 2g, citric acid tri-amonia 1g, sodium acetate 5g, magnesium sulfate 0.8g, manganese sulfate 0.25g, Tween 80 1ml, pH value 6.2-6.4, be settled to 1000ml with distilled water, 115-120 DEG C of sterilizing 30min.
This area bacillus cereus and Lactobacillus plantarum isolation medium select common MRS culture medium: tryptone 10g, beef extract 10g, dibasic ammonium citrate 2g, sodium acetate 5g, dusty yeast 5g, glucose 5g, potassium dihydrogen phosphate 2g, Tween 80 1.0ml, calcium carbonate 20g, magnesium sulfate 0.58g, manganese sulfate 0.25g, water 1000ml, adjust pH 6.2-6.5,115 DEG C of sterilizing 20min.
The modified MRS culture medium that bacillus cereus of the present invention and Lactobacillus plantarum adopt: add α-methylglucoside 10g, potassium sorbate 1g on MRS medium base, adjust pH 6.2-6.5,115 DEG C of sterilizing 20min; Culture medium prescription after change is, dusty yeast 3g/L, peptone 5g/L, beef extract 2g/L, MnSO 40.005g/L, NaCl 2g/L, K 2hPO 43g/L, MgSO 40.02g/L, this medium base carries out the optimization of fermentation condition, show that the condition of culture after optimization is: temperature 40 DEG C, initial pH value is 7.0, rotating speed 210r/min, and liquid amount is 30mL (triangular flask of 250mL), inoculum concentration is 6% (v/v), fermentation time 48h.
Embodiment two: the cotton dregs compound micro-ecological preparation being suitable for aquaculture
The present invention specifically provides a kind of cotton dregs compound micro-ecological preparation being suitable for aquaculture, by the agent of cotton dregs complex group with deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 60%, sunflower seed dregs 10%, the dish dregs of rice 13%, the composite bacteria agent capable of safflower seed meal 12% and the grape pip dregs of rice 5% and 1.5% prepares the agent of cotton dregs complex group by solid state fermentation, and composite bacteria agent capable mixes acquisition by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum according to the ratio of weight ratio 3:5:2; Deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix; To mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
Embodiment three: the cotton dregs compound micro-ecological preparation being suitable for aquaculture
The present invention specifically provides a kind of cotton dregs compound micro-ecological preparation being suitable for aquaculture, by the agent of cotton dregs complex group with deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 70%, sunflower seed dregs 8%, the dish dregs of rice 15%, the composite bacteria agent capable of safflower seed meal 5% and the grape pip dregs of rice 2% and 1.5% prepares the agent of cotton dregs complex group by solid state fermentation, and composite bacteria agent capable mixes acquisition by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum according to the ratio of weight ratio 3:5:2; Deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix; To mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
Embodiment four: the cotton dregs compound micro-ecological preparation being suitable for aquaculture
The present invention specifically provides a kind of cotton dregs compound micro-ecological preparation being suitable for aquaculture, by the agent of cotton dregs complex group with deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 65%, sunflower seed dregs 9%, the dish dregs of rice 14%, the composite bacteria agent capable of safflower seed meal 8.5% and the grape pip dregs of rice 3.5% and 1.5% prepares the agent of cotton dregs complex group by solid state fermentation, and composite bacteria agent capable mixes acquisition by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum according to the ratio of weight ratio 3:5:2; Deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix; Mix according to the cotton dregs complex group agent of every 500g form cotton dregs compound micro-ecological preparation with the mixed bacteria pulvis of delivering directly of 500 kilograms.
Embodiment five: the preparation being suitable for the cotton dregs compound micro-ecological preparation of aquaculture
The present invention specifically provides a kind of preparation method being suitable for the cotton dregs compound micro-ecological preparation of aquaculture, prepares especially by following technique, and % is according to weight percent meter, and concrete preparation method's step is as follows:
(1) pretreatment of raw material: by raw material cotton dregs 60%-70%, sunflower seed dregs 8%-10%, dish dregs of rice 15%-13%, safflower seed meal 5%-12% and grape pip dregs of rice 2%-5%, mix in proportion and pulverize, and crosses 40 mesh sieves; Add water according to the solid-liquid ratio of weight ratio 1:0.8, be uniformly mixed acquisition raw material.
(2) acquisition of the activation of bacterial classification, cultivation and microbial inoculum: respectively bacillus coagulans, candida utili and the aspergillus niger selected are carried out test tube slant activation culture respectively, and the bacterial classification activated through test tube slant is transferred in shaking flask cultivates, obtain three kinds of bacillus coagulanses, candida utili and black-koji mould microbial inoculums respectively.
(3) preparation of composite bacteria: bacillus coagulans microbial inoculum, candida utili microbial inoculum and the aspergillus niger microbial inoculum ratio according to weight ratio 3:5:2 is mixed, is the composite bacteria agent capable of preparation.
(4) bacterium is connect: the composite bacteria agent capable prepared by step (3) is linked in raw material prepared by step (1) by the inoculative proportion with 1.5%.
(5) solid fermentation: adopt common Zymolysis Equipment, raw material prepared by whipping step (1) and the composite bacteria agent capable that step (3) is added, make it to mix, and is 30 DEG C-35 DEG C through controlling fermentation temperature, pH nature, continuous solid-state fermentation 48h.
(6) dry, granulate: the raw material that above-mentioned steps (5) fermentation is got ready through granulation, dry to moisture be 12% acquisition cotton dregs complex group agent.
(7) bacillus cereus and Lactobacillus plantarum is selected to obtain pulvis through the activation of bacterial classification, the collection of thalline, the freeze drying of bacterium mud respectively:
The activation of bacterial classification: the bacillus cereus selected and Lactobacillus plantarum are carried out first time by the MRS liquid culture medium of 100ml sterilizing respectively and activates, activation temperature 37 DEG C, 24h rear plate detects purity and viable count, detect up to standard after more above-mentioned 100ml bacterium liquid is all proceeded to 400ml sterilizing MRS liquid culture medium carry out second time activation, 24h rear plate detects purity and viable count, detect up to standard after go in fermentation tank and carry out expansions cultivation 48h.
Microorganism collection: utilize the bacterium liquid of ultracentrifuge to the bacillus cereus after fermentation and Lactobacillus plantarum to carry out centrifugal respectively, be centrifugal 10min under the condition of 4000-6000r/min at rotating speed, go supernatant to collect deposit.
Bacterium mud freeze drying: the deposit of the bacillus cereus of collecting respectively and Lactobacillus plantarum is added to composite protective agent, with oscillator mixing, makes bacteria suspension; Contained in freezing plate again, after first putting 4 DEG C of refrigerator balance 30min subsequently, move into-30 DEG C of refrigerator 60min, then-80 DEG C of refrigerator 60min are put, freeze drying 14h on frozen vacuum dryer again, sealing is stored in 4 DEG C of refrigerators, prepares bacillus cereus pulvis and Lactobacillus plantarum pulvis respectively.Experimentally design, vacuum freeze drying condition setting freeze-drying instrument cavity temperature is-60 DEG C-70 DEG C, and pressure is 0.006Pa, and the time is 14h; Composite protective agent adopts skimmed milk power 7.06%, maltose 6.46% and sodium glutamate 6.70%.
(8) allocate: bacillus cereus pulvis above-mentioned steps obtained respectively and Lactobacillus plantarum pulvis form according to weight ratio 1:1 delivers directly mixed bacteria pulvis.
(9) finished product: the cotton dregs complex group agent of selecting above-mentioned steps (6) to prepare and step (8) prepare delivers directly mixed bacteria pulvis, to mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
The cotton dregs compound micro-ecological preparation good palatability prepared by above-mentioned technique, can promote the appetite of fish; There is good formability and resistance to water, ensure that the stability of cotton dregs compound micro-ecological preparation in water, reduce the pollution of water body; Meanwhile, the digestibility in fish body is high, promotes the growth of fish body, is with a wide range of applications.
The cotton dregs that the present invention adopts, sunflower seed dregs, the dish dregs of rice, the pulverizing of safflower seed meal and the grape pip dregs of rice, add water, the pretreatment such as mixing, the activation of bacterial classification, fermentation, although the technological means such as the freeze drying of bacterial classification be unable to do without support and the inspiration of prior art, but the material ratio of cotton dregs compound micro-ecological preparation in zymotechnique, solid-liquid ratio, , the selection of fermented bacterium and deliver directly the selection of mixed bacteria, preparation, for determining that the quality of cotton dregs compound micro-ecological preparation all has important effect, also be deciding factor, technological means and the instrument of not talkative employing are prior aries, and then negate the novelty that entirety provided by the invention prepares cotton dregs compound micro-ecological preparation, existing all technological inventions and innovation all be unable to do without support and the basis of prior art.Visible, each technical factor in cotton dregs compound micro-ecological preparation preparation technology provided by the invention and technical parameter are all through comprehensive consideration and design, in the preparation method of cotton dregs compound micro-ecological preparation provided by the invention, each sport technique segment has one up and down, any technical step is indispensable, tight indivisible feature, can not approbate and reprobate dismemberent technical step wherein.
Embodiment six: the preparation being suitable for the cotton dregs compound micro-ecological preparation of aquaculture
By raw material cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and the grape pip dregs of rice, mix in proportion and pulverize, crossing 40 mesh sieves; Add water according to the solid-liquid ratio of weight ratio 1:0.8, be uniformly mixed acquisition raw material; Respectively bacillus coagulans, candida utili and aspergillus niger strain carried out activation culture and obtain corresponding three kinds of microbial inoculums, and obtain composite bacteria agent capable according to the ratio mixing of weight ratio 3:5:2, be linked in the raw material of preparation with the inoculative proportion composite bacteria agent capable of 1.5%; Adopt solid-state fermentation process, fermentation temperature is 30 DEG C-35 DEG C, pH nature, continuous solid-state fermentation 48h-60h, and through drying, granulating, obtains the cotton dregs complex group agent that moisture is 12%; Select bacillus cereus and Lactobacillus plantarum to obtain corresponding two kinds of pulvis through freeze drying respectively, form acquisition according to weight ratio 1:1 and deliver directly mixed bacteria pulvis; The cotton dregs complex group agent delivering directly mixed bacteria pulvis and 500 kilograms according to every 500g is mixed with and obtains cotton dregs compound micro-ecological preparation.
After feed protein is ingested by fish body, in the alimentary canal of fish under the effect of various digestive ferment, could must be absorbed by fish body after breaks down into amino acids.In feed, the content of protein and effective energy value play vital effect to growing of fish.Meanwhile, vitamin is requisite special dietary material in the metabolism of fish substance in vivo.Cottonseed meal matter content is higher, but form not ideal, arginine content is up to 3.6%-3.8%, and lysine content only has 1.3%-l.5%, for the half of Soybean Meal, methionine is also not enough, and about 0.4%, the utilization rate of lysine is poor, phosphorous many in mineral matter, but belong to phytate phosphorus, utilization rate is low more, therefore need to add the raw materials such as the dish dregs of rice, the grape pip dregs of rice, the protein content of sunflower seed dregs is very large according to shelling degree difference, at about 24%-38%, the amino acid compositing characteristic of sunflower seed dregs is that lysine content is not enough, at about 1.1%-1.2%, lower than cotton dregs, methionine content is higher, about about 0.6%-0.7%, higher than dregs of beans, cotton dregs and peanut meal, the B family vitamin rich content of sunflower seed dregs, Nicotinic Acid Content reaches 200mg/kg, be 5 times of dregs of beans, but lack carrotene, calcium in mineral matter, phosphorus, iron, zinc, copper content are also higher, therefore, need to add the raw material such as cotton dregs, the dish dregs of rice, containing abundant lysine in the dish dregs of rice, macro-and trace-element, wherein the content of calcium, selenium, iron, magnesium, manganese, zinc is higher than dregs of beans, phosphorus content is 2 times of dregs of beans, it is also containing abundant sulfur-containing amino acid simultaneously, this just dregs of beans lack, so balance and complementation can be played when it and dregs of beans share, in safflower seed meal, protein content is 20%-60%, and its arginine content is up to 1.2%-3.7%, and containing abundant vitamin and mineral matter, the many phosphorus of calcium is few, and containing multiple polyphenoils, the grape pip dregs of rice are rich in a large amount of polyphenols, linoleic acid, amino acid, multivitamin, crude fat, trace element, wherein, polyphenols procyanidins is main matter, there is superpower oxidation resistance, it is 50 times of vitamin E, it is ascorbic 20 times, free radical too much in its energy purged body, its oxidation resistance has obtained internationally recognized, at present, the grape pip dregs of rice have been widely used in feed, not only save a large amount of grain, and significantly improve economic benefit, therefore, the interpolation of the grape pip dregs of rice has important effect for the growth of fish body.The present invention by fully analyzing the nutritive peculiarity of these five kinds of dregs of rice class materials of cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and the grape pip dregs of rice, and according to the growth needs of fish body, prepares the agent of cotton dregs complex group by fermentable effect.Meanwhile, deliver directly mixed bacteria pulvis according to factor preparations such as the digestion of fish body, it is made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis.By the agent of cotton dregs complex group with deliver directly mixed bacteria pulvis and be mixed with cotton dregs compound micro-ecological preparation.
Embodiment seven: the mensuration of cotton dregs compound micro-ecological preparation water stability
(1) experimentation
Random selecting 20 particle test feed is placed in the water surface gently, makes its free settling, and presses stopwatch, records the time t of its free settling, with the height h of ruler measurement liquid level and bottom.
Rate of settling v=h/t
In formula, h is liquid level (cm); T is for falling heavy time (s)
(2) experimental result
Be statically placed in water body by cotton dregs compound micro-ecological preparation, it all floats on water surface, and after soaking 15min, feed granules starts to sink, and final nearly half feed is in water body upper strata.Density because of cotton dregs compound micro-ecological preparation is less than water body density, may float on water surface, along with feed water absorption increases, engenders sinkage, changes similar to feed water absorption character.Soak about 15min, water absorption is tending towards saturated, feed granules sinking increasing number.Calculated settlement speed is 1cm/s.Feed granules suspends or dipping and heaving in water body, shows because of water absorption course, and feed is in the state tended to balance.Still have nearly half particle to be in water body upper strata, illustrate that feed granules keeps poised state in hydrostatic, cotton dregs compound micro-ecological preparation is strong at water stability.
Embodiment eight: the fish application test of cotton dregs compound micro-ecological preparation
1, subjects
Select pond rearing adult fish of cyprinoid then.After adult fish is bought back, be placed in cement pit, conveniently feeding and management method, feed 7d with adult fish of cyprinoid feed.On an empty stomach after 24h, weighed, the carp of selection body weight, uniformity of type is placed in test net cage, about average weight 710g.Arrange test group and control group two groups, the cotton dregs compound micro-ecological preparation that test group selects above-described embodiment to provide, the fish that control group selects market common are suitable for feed formulations, and often group 3 is parallel, totally 6 groups, often organize Random assignment test fish 30 tail, amount to 180 tails.
2, feeding method
Culture experiment starts from early October, to terminating by the end of November, 8 weeks by a definite date, amounts to 56d.Before on-test, the test carp saline solution of 3% soaks 10-15min, carries out routine disinfection, and adult fish of cyprinoid feed of feeding domestication 1 week, to conform.Hungry 24h, weighs to the fish in each test tank.Test fish respectively 10 of every day time, 14 time, 18 time throw something and feed, day feeding rate 2-3%, be advisable, or 0.5h has eaten to be just satiated with food.Carry out sampling every 2 weeks to each test group fish to weigh, adjustment day feeding volume, and adjust feeding volume in time according to water temperature and growing state.
During cultivation, every morning and afternoon respectively measure water temperature once, and observation fish body is travelling judges water body dissolved oxygen, and every day measures pH value once with pH test paper, every day soil pick-up once, pull dead fingerling in time out, carry out weighing, record, the outward appearance of dead fish body and internal organs are described simultaneously.
3, the testing index of fish body
(1) fish bulk-growth index
The speed of growth of fish body is according to specific growth rate, rate of body weight gain index expression.Stop eating after 24h and weigh to test fish body, the test fish of each aquaculture net cage carries out overall weighing, and indices is according to following formula.
Specific growth rate (SGR, %/d)=[ln (W1)-ln (W0)]/t × l00
Rate of body weight gain (%)=(W1-W0)/W0 × l00
Wherein Wl, W0 are respectively fish body and raise last heavy and initial weight, and t is test number of days.
(2) fish volume morphing index
During off-test, randomly draw 5 tail fishes and carry out the long measurement of body weight, body in each aquaculture net cage, then solution takes internal organ and weighs.Calculate coefficient of condition and internal organ index index.
Coefficient of condition (%)=body weight (g)/body long (cm)
Internal organ index (%)=internal organ heavy (g)/heavy (g) x100 of full fish
(3) Major Components of fish body muscle
Fish body muscle is freezing sample, gets 5 tail fishes in each aquaculture net cage respectively, in refrigerator-freezer after freezing 1 week, gets back white muscle, measures, and testing index is the content of moisture, crude protein, crude fat, coarse ash.
(4) health indicator
During off-test, the internal organs of the 5 tail fishes dissected in each aquaculture net cage are separated further, get kidney respectively, hepatopancrease, spleen weigh, calculate index of correlation.Comprise hepatopancreatic cell, index and spleen index and head kidney index.
4, result of the test
(1) speed of growth
Table 1 is the counterpoise of group fish body, rate of body weight gain and specific growth rate respectively
First counterpoise/g End counterpoise/g Rate of body weight gain/% Specific growth rate/%
A1 716.7 924.1 28.94 0.45
A2 714.0 926.8 29.80 0.46
A3 710.6 927.2 30.48 0.48
B1 712.3 798.4 12.08 0.20
B2 716.4 818.9 14.31 0.24
B3 710.4 792.5 11.55 0.20
Test the fish specific growth rate of 8 weeks and rate of body weight gain as shown in table 1.The specific growth rate of test group A and rate of body weight gain are significantly higher than control group B, reach 0.46% and 29.74% respectively, improve 0.25% and 17.09% than control group.Above-mentioned analysis shows that experimental group cotton dregs provided by the invention compound micro-ecological preparation feed demonstrates good growth result to carp, and is better than corresponding control group feed.
(2) fish volume morphing index: test group A coefficient of condition with higher than control group B, reach 30.72g/cm, than control group height 3.61g/cm.And the internal organ index of test group A is significantly lower than control group B, reach 14.49% and 16.14% respectively.Test group coefficient of condition is high, the low explanation test group A of internal organ index compared with control group, the quality of unit length fish body is large, fish body grossness, and fish body trunk ratio is large, and edible part is many.
(3) muscle basis is analyzed: as shown in Table 2, compared with control group, test group moisture reduces, but difference is not significantly (P > 0.05), test group fat content is all lower than control group, and difference is not significantly (P > 0.05).In test group and control group fish body muscle, content of ashes is close, there is no significant difference.Test group adopts the crude protein content of cotton dregs compound micro-ecological preparation provided by the invention to be significantly higher than test group fish body crude protein content (P < 0.05).
Table 2 is group fish body muscle basis respectively
Moisture/% Crude fat/% Crude protein/% Ash content/%
A1 77.54±0.12 2.05±0.53 20.01±0.08 1.19±0.21
A2 77.04±0.42 3.42±0.28 20.30±0.34 1.20±0.22
A3 77.24±0.26 3.77±0.20 20.14±0.23 1.08±0.18
B1 78.71±0.18 3.83±0.87 19.37±0.20 1.05±0.08
B2 76.63±0.66 3.91±0.45 19.59±0.07 1.35±0.14
B3 78.75±0.03 3.95±0.57 19.20±0.01 1.15±0.20
(4) health indicator
Table 3 is group fish body hepatopancrease, kidney, index and spleen index respectively
Hepatopancreatic cell Renal index Index and spleen index
A1 2.20±0.39 0.68±0.02 0.26±0.02
A2 2.27±0.17 0.60±0.01 0.26±0.05
A3 2.29±0.04 0.53±0.06 0.30±0.01
B1 2.26±0.02 0.60±0.04 0.33±0.03
B2 1.77±0.03 0.61±0.01 0.28±0.02
B3 1.61±0.26 0.55±0.03 0.33±0.01
Liver pancreas is the important metabolic organ of aquatic livestock, and have the function of dispelling toxicity, detoxicating, liver pancreas index can reflect the fish such as hepatomegaly, fatty liver body physiological problem.Test group compared with control group, test group A2, A3 hepatopancreatic cell all higher than control group, test group with contrast group difference significantly (P > 0.05).Duration of test, does not find to adopt the above-mentioned cotton dregs compound micro-ecological preparation provided of the present invention to cause adverse effect to test Hepatopancreas of Cyprinus Carpio.After off-test, the liver pancreas color that fish body is dissected is normal peony, and hepatopancreatic cell shows that the liver pancreas of each test group fish body is not affected.Test group is close with control group renal index, a little higher than control group, and difference is not significantly (P > 0.05).Tentatively determine that cotton dregs compound micro-ecological preparation does not impact immune cell.Spleen is the important blood forming organ of body, filters blood and also participates in organism immune response.Test group index and spleen index is 0.27 a little less than control group index and spleen index 0.31, and difference is not remarkable.Test feed and commercial feed affect close on spleen, adopt cotton dregs compound micro-ecological preparation provided by the invention not impact spleen.
Conclusion: adopt cotton dregs compound micro-ecological preparation provided by the invention to demonstrate good effect in carp growth, the quality of the carp unit length fish body of raising with cotton dregs compound micro-ecological preparation is large, fish body grossness, and fish body trunk ratio is large, edible part is many, is better than corresponding commercial feed.The flesh quality that flesh quality and commercial feed are fed is close, is even better than commercial feed.Adopt cotton dregs compound micro-ecological preparation provided by the invention not impact fish body health, reach commercial feed level, harmful effect is not caused to the physiological health of fish body.In a word, adopt cotton dregs compound micro-ecological preparation provided by the invention close with the effect of commercial feed in carp raising, be even better than commercial feed in some respects.
By a kind of cotton dregs compound micro-ecological preparation and Synthesis and applications being suitable for aquaculture that the various embodiments described above provide, cotton dregs, sunflower seed dregs, the dish dregs of rice, safflower seed meal and grape pip dregs of rice mixture is utilized to be raw material, microbial fermentation technology is utilized to prepare the agent of cotton dregs complex group, prepare direct putting type probiotics by freeze drying technology, match by utilizing the agent of cotton dregs complex group and direct putting type probiotics and prepare the novel cotton dregs compound micro-ecological preparation being applicable to aquaculture.The technical problem solved be to overcome prior art defect, make full use of the protein raw materials such as cotton dregs, sunflower seed dregs, the sour-sweet fragrance of cotton dregs compound micro-ecological preparation smell of preparation, good palatability, can promote the appetite of fish; There is good formability and resistance to water, ensure that the stability of cotton dregs mixed feed in water, reduce the pollution of water body; Meanwhile, the digestibility in fish body is high, is with a wide range of applications.
Above-described embodiment is only for example of the present invention is clearly described, and the restriction not to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without the need to also giving all embodiments.And apparent change extended thus or variation are still among protection scope of the present invention.

Claims (6)

1. one kind is suitable for the cotton dregs compound micro-ecological preparation of aquaculture, it is characterized in that, % by weight percentage, cotton dregs compound micro-ecological preparation proportioning is by the agent of cotton dregs complex group and deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 60%-70%, sunflower seed dregs 8%-10%, dish dregs of rice 15%-13%, the composite bacteria agent capable of safflower seed meal 5%-12% and grape pip dregs of rice 2%-5% and interpolation 1.5%, the agent of cotton dregs complex group is prepared by solid state fermentation, composite bacteria agent capable is by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum obtain according to the ratio mixing of weight ratio 3:5:2, deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix, to mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
2. a kind of cotton dregs compound micro-ecological preparation being suitable for aquaculture as claimed in claim 1, it is characterized in that, by weight percentage, cotton dregs compound micro-ecological preparation proportioning is by the agent of cotton dregs complex group and deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 60%, sunflower seed dregs 10%, the dish dregs of rice 13%, the composite bacteria agent capable of safflower seed meal 12% and the grape pip dregs of rice 5% and interpolation 1.5%, the agent of cotton dregs complex group is prepared by solid state fermentation, composite bacteria agent capable is by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum obtain according to the ratio mixing of weight ratio 3:5:2, deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix, to mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
3. a kind of cotton dregs compound micro-ecological preparation being suitable for aquaculture as claimed in claim 1, it is characterized in that, by weight percentage, cotton dregs compound micro-ecological preparation proportioning is by the agent of cotton dregs complex group and deliver directly mixed bacteria pulvis and form, the agent of cotton dregs complex group contains cotton dregs 70%, sunflower seed dregs 8%, the dish dregs of rice 15%, the composite bacteria agent capable of safflower seed meal 5% and the grape pip dregs of rice 2% and interpolation 1.5%, the agent of cotton dregs complex group is prepared by solid state fermentation, composite bacteria agent capable is by bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum obtain according to the ratio mixing of weight ratio 3:5:2, deliver directly mixed bacteria pulvis to be made up of according to weight ratio 1:1 bacillus cereus pulvis and Lactobacillus plantarum pulvis and to mix, to mix with the cotton dregs complex group agent of 500 kilograms form cotton dregs compound micro-ecological preparation according to the mixed bacteria pulvis of delivering directly of every 500g.
4. a kind of preparation method being suitable for the cotton dregs compound micro-ecological preparation of aquaculture as described in claims 1 to 3 any one, is characterized in that, % is according to weight percent meter, and the concrete preparation process of cotton dregs compound micro-ecological preparation is as follows:
(1) pretreatment of raw material: by raw material cotton dregs 60%-70%, sunflower seed dregs 8%-10%, dish dregs of rice 15%-13%, safflower seed meal 5%-12% and grape pip dregs of rice 2%-5%, mix in proportion and pulverize, and crosses 40 mesh sieves; Add water according to the solid-liquid ratio of weight ratio 1:0.8, be uniformly mixed acquisition raw material;
(2) acquisition of the activation of bacterial classification, cultivation and microbial inoculum: respectively bacillus coagulans, candida utili and aspergillus niger strain are carried out test tube slant activation culture, and the bacterial classification activated through test tube slant is transferred in shaking flask cultivates, obtain bacillus coagulans microbial inoculum, candida utili microbial inoculum and aspergillus niger microbial inoculum respectively;
(3) preparation of composite bacteria agent capable: bacillus coagulans microbial inoculum, candida utili microbial inoculum and the aspergillus niger microbial inoculum ratio according to weight ratio 3:5:2 is mixed, is and prepares composite bacteria agent capable;
(4) bacterium is connect: the composite bacteria agent capable prepared by step (3) is linked in raw material prepared by step (1) by the inoculative proportion with 1.5%;
(5) solid fermentation: adopt common Zymolysis Equipment, raw material prepared by whipping step (1) and the composite bacteria agent capable that step (3) is added, make it to mix, and is 30 DEG C-35 DEG C through controlling fermentation temperature, pH nature, continuous solid-state fermentation 48h-60h;
(6) dry, granulate: the raw material that above-mentioned steps (5) fermentation is got ready through granulation, dry to moisture be 12% acquisition cotton dregs complex group agent;
(7) preparation of bacterial classification pulvis: select bacillus cereus and Lactobacillus plantarum to obtain pulvis through the activation of bacterial classification, the collection of thalline, the freeze drying of bacterium mud respectively;
(8) allocate: bacillus cereus pulvis above-mentioned steps obtained respectively and Lactobacillus plantarum pulvis form according to weight ratio 1:1 delivers directly mixed bacteria pulvis;
(9) finished product: the cotton dregs complex group agent of selecting above-mentioned steps (6) to prepare and step (8) prepare to be delivered directly mixed bacteria pulvis and be mixed with and obtain cotton dregs compound micro-ecological preparations.
5. a kind of preparation method being suitable for the cotton dregs compound micro-ecological preparation of aquaculture as claimed in claim 4, the preparation of described bacterial classification pulvis, bacillus cereus and Lactobacillus plantarum is selected to obtain pulvis through the activation of bacterial classification, the collection of thalline, the freeze drying of bacterium mud respectively, it is characterized in that, the preparation concrete steps of bacterial classification pulvis are as follows:
(1) activation of bacterial classification: the bacillus cereus selected and Lactobacillus plantarum are carried out first time by the MRS liquid culture medium of 100ml sterilizing respectively and activates, activation temperature 37 DEG C, 24h rear plate detects purity and viable count, detect up to standard after more above-mentioned 100ml bacterium liquid is all proceeded to 400ml sterilizing MRS liquid culture medium carry out second time activation, 24h rear plate detects purity and viable count, detect up to standard after go in fermentation tank and carry out expansions cultivation 48h;
(2) microorganism collection: utilize the bacterium liquid of ultracentrifuge to the bacillus cereus after fermentation and Lactobacillus plantarum to carry out centrifugal respectively, be centrifugal 10min under the condition of 4000-6000r/min at rotating speed, go supernatant to collect deposit;
(3) bacterium mud freeze drying: the deposit of the bacillus cereus of collecting respectively and Lactobacillus plantarum is added to composite protective agent, with oscillator mixing, makes bacteria suspension; Contained in freezing plate again, after first putting 4 DEG C of refrigerator balance 30min subsequently, move into-30 DEG C of refrigerator 60min, then put-80 DEG C of refrigerator 60min, freeze drying 14h on frozen vacuum dryer again, prepares bacillus cereus pulvis and Lactobacillus plantarum pulvis respectively; Experimentally design, vacuum freeze drying condition setting freeze-drying instrument cavity temperature is-60 DEG C-70 DEG C, and pressure is 0.006Pa, and the time is 14h; Composite protective agent adopts skimmed milk power 7.06%, maltose 6.46% and sodium glutamate 6.7%.
6. the application of cotton dregs compound micro-ecological preparation in aquaculture as described in claim 1,2 and 3 any one claim.
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