CN108185213A - The preparation method of loach seedling nutrition enhancer - Google Patents
The preparation method of loach seedling nutrition enhancer Download PDFInfo
- Publication number
- CN108185213A CN108185213A CN201711238844.XA CN201711238844A CN108185213A CN 108185213 A CN108185213 A CN 108185213A CN 201711238844 A CN201711238844 A CN 201711238844A CN 108185213 A CN108185213 A CN 108185213A
- Authority
- CN
- China
- Prior art keywords
- parts
- preparation
- meretrix
- loach seedling
- fish oil
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- 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
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- 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
- A23K20/105—Aliphatic or alicyclic compounds
-
- 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
- A23K20/111—Aromatic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/179—Colouring agents, e.g. pigmenting or dyeing agents
-
- 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
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/22—Compounds of alkali metals
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Animal Husbandry (AREA)
- Food Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Marine Sciences & Fisheries (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Botany (AREA)
- Mycology (AREA)
- Sustainable Development (AREA)
- Inorganic Chemistry (AREA)
- Insects & Arthropods (AREA)
- Birds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses the preparation method of loach seedling nutrition enhancer, including：Fish meal fermentation, the preparation of Meretrix meretrix Glycopeptide, the preparation of fish oil micro-capsule, the preparation of Chinese medicine preparation, compounding, nutrition enhancer by following parts by weight material composition：40 50 parts of fermentation fish meals, 10 15 parts of Meretrix meretrix Glycopeptides, 20 30 parts of fish oil micro-capsules, 15 20 parts of Chinese medicine preparations, 20 22 portions of salt, 56 parts of Yolk antibodies, 24 parts of sodium butyrates, 56 parts of carrotene, 23 parts of sulfa powders.It has the beneficial effect that：Reinforcing agent cooperation conventional feed uses, loach seedling can be increased to ingest, also there is stronger antioxidation, free radical extra in loach seedling body can be removed, while reinforcing agent also has bacteriostasis antibiosis activity, can effectively enhance the immunocompetence of loach seedling, enhance loach seedling constitution and vigor, eyesight, the anti-stress ability of loach seedling are improved, raising speed is improved, reduces aquaculture cost.
Description
Technical field
The present invention relates to the nutrition enhancer field of loach seedling, more particularly, to the preparation of loach seedling nutrition enhancer
Method.
Technical background
Loach belongs to Cobitidae.Body is small, elongated, only three, four cun.And its bodily form is justified, body is short, subcutaneously there is ramentum, color
It is green black, it has been stained with the mucus of itself from head to foot, thus satiny can not hold.Leading portion is slightly cylindrical.Rear portion is flat-sided, abdomen circle, head
It is small, mouth is small, the next, the shape of a hoof.Eye is small, and nothing is pierced now.It must be 5 pairs.Squama is extremely tiny, round, is embedded in subcutaneous.Body back and both sides
Grey black, it is all there are many small black patches, also there are many black splotch on head and each fin, the spot on dorsal fin and tail fin film
It is arranged in rows, caudal peduncle base portion has an apparent blackspot.Other each fin canescence.Loach be distributed widely in Asia bank China,
The ground such as Japan, Korea, Russia and India.In distribution in China various regions.South distribution is more, and the north is not common.Whole year can all adopt
It receives, summer is most, can using fresh herb or drying use after loach captures.Edible is used as medicine.Loach is referred to as " ginseng in water ", lives in
Hu Chi is a kind of very high fish of nutritive value, it and other fishes differ, no matter appearance, the bodily form, life habit is all different,
It is a kind of special bullhead.
The prior art discloses loach specific complex as authorized the Chinese invention patent that public platform is 104222680 B of CN
Immunopotentiator and preparation method thereof, the composite immune reinforcing agent are made of the component of following parts by weight：5-10 parts of garlic, rheum officinale
5-15 parts of extract, 1-5 parts of hickory nut pericarp crude extract, 4-6 parts of basyleave, 1-5 parts of Poplar leaves, floats at 1-5 parts of forsythia suspense extraction
5-10 parts of duckweed extract, 10-15 parts of hawthorn, 10-20 parts of salt, the additive in the invention are used cooperatively with conventional feed, can
Enhance loach immunity of organisms, improve resistance.But the component in the composite immune reinforcing agent in the inventive method is mostly to plant
Object or plant extracts can not provide required nutrition enhancing material for loach.
Invention content
The purpose of the present invention is to provide the preparation methods of loach seedling nutrition enhancer, and preparation method is simple and practicable, mud
Loach seedling nutrition enhancer can significantly increase the immunocompetence of loach seedling, anti-stress ability, can greatly improve raising effect
Rate reduces aquaculture cost.
Used microorganism and its bought enterprise are in the present invention：Bacillus subtilis, the Cangzhou City macro animal of benefit protect
Jian Pin Co., Ltds；Lactobacillus acidophilus, Shaanxi Sen Fu natural products Co., Ltd.
The problem of present invention in background technology for mentioning, the technical solution taken is：Loach seedling nutrition enhancer
Preparation method, including：Fish meal fermentation, the preparation of Meretrix meretrix Glycopeptide, the preparation of fish oil micro-capsule, the preparation of Chinese medicine preparation, compounding, tool
Body includes the following steps：
Fish meal ferments：Take fish meal with distilled water according to solid-liquid ratio 1:The ratio of 2-3 is uniformly mixed, and is inoculated with the mixing into 3.5-4.5%
Strain, fermentation time 12-16 hours at a temperature of 32-33 DEG C, low temperature drying after fermentation obtains fermentation fish meal；This condition issues
The small peptide content of ferment fish meal reaches 29.8%, and Volatile Base Nitrogen is only 40.3mg/100g, and the content of limiting amino acid is promoted
More than 30.2%, the more common fish meal of total antioxidant capacity promotes more than 70%, has and removes the huge of excessive free radicals in organism
Potentiality；In addition, in fermentation fish meal, the content of the limiting amino acids such as lysine, threonine, methionine, histidine, which has, to be shown
It writes and improves, disclosure satisfy that demand of the loach seedling to limiting amino acid；
The preparation of Meretrix meretrix Glycopeptide：Clam is taken to obtain clam dry powder after spitting sand, decladding, freeze-drying, crushing, by solid-liquid ratio 1:6-7 is molten
It in distilled water and is homogenized, is centrifuged 30-45 minutes in 5-6 DEG C, 500-600r/min, collect supernatant, then through Hollow Fiber Ultrafiltration
Column ultrafiltration, ultrafiltration column molecular cut off are 8 × 104, ultrafiltration collect small-molecular-weight ultrafiltrate, 1/5-2/5 is added in ultrafiltrate
The chloroform of volume, the n-butanol of 1/25-1/20 volumes add 1,3, the 5- trimethoxy-benzenes of n-butanol weight 2-6%, extraction
2-3 times；Extract liquor is dialysed, is centrifuged, be dried in vacuo after obtain Meretrix meretrix Glycopeptide crude product, centrifuging and taking supernatant after crude product is water-soluble, then with
0.2mol/L, 0.4mol/L, 0.6mol/L, 0.8mol/L, 1.0mol/L sodium chloride solution gradient elution, obtain Meretrix meretrix Glycopeptide；1,
With n-butanol the effects that hydrogen bond is connect can occur for 3,5- trimethoxy-benzenes, and then greatly enhance the extraction efficiency of system, improve
The purity and yield of Meretrix meretrix Glycopeptide, the purity of gained Meretrix meretrix Glycopeptide can reach 97%, sugared and each 54% He of protein content
42%, Meretrix meretrix Glycopeptide has excellent antitumor activity, bacteriostasis antibiosis activity, can effectively enhance the immunity energy of loach seedling
Power enhances loach seedling constitution and vigor；
The preparation of fish oil micro-capsule：0.46-0.56 parts of Tween-80 mixing are added in 100-120 parts by weight gumwaters
Uniformly, 12-14 parts of fish oil are added in, interval ultrasound 5-6 minutes, supersonic frequency 25-30kHz, ultrasonic density are 0.35-0.45W/
cm2, second mode for ultrasound 2 seconds, pause 4 seconds, form uniform emulsion；300-400 parts of chitosan solutions are added drop-wise to dropwise
It is 4.5-5.0 with hydrochloric acid and sodium hydroxide solution adjustment pH, in 18-20 DEG C in the Arabic gum fish oil emulsion being stirred continuously
Lower stirring 30-45 minutes；Stratification, condensation product enrichment phase are that chitosan-Arabic gum agglomerates fish oil micro-capsule again；Fish oil
In rich in omega-3 polyunsaturated fatty acids EPA and DHA, having reduces serum cholesterol and low-density lipoprotein content, improves and regard
The different physiological roles such as power, anticoagulation, anticancer also have the function of to improve organism lipid structure；But meanwhile in EPA and DHA
It is extremely sensitive to light, heat and oxygen etc. containing multiple double bonds, it needs to be protected from light, low temperature and sealing storage and transportation, largely limits fish
Application of the oil in industrial circle；Chitosan-Arabic gum agglomerates fish oil micro-capsule in regular spherical shape again, favorable dispersibility,
Particle diameter distribution is uniform, and average grain diameter is 16-20 μm, and the utilization rate of embedding rate and wall material to fish oil is respectively 75.33% He
81.01%, can be preferable hidden by fish oil, the influence of its light, heat, oxygen is reduced, improves fish oil utilization rate；
The preparation of Chinese medicine preparation：Weigh 10-12 parts of radix scutellariaes, 3-4 portions of Chinese galls, 3-4 parts of Enteromorphas, 10-15 portions of cloves, 2-5 parts of jade
Bamboo, 5-8 part Radix Astragali, 10-12 parts of Herba Epimedii smash it through 120-160 mesh sieve, traditional Chinese medicine powder are taken to add water to cook 60-90 minutes, natural
Take supernatant concentration to ointment up to Chinese medicine preparation after cooling；Chinese medicine preparation safety, low toxicity, low-residual, are not likely to produce drug resistance,
Tonifying spleen, stomach invigorating, tonifying Qi, solid table, adjust metabolism, enhance spleen and stomach function, can reduce stress reaction, the incidence of loach seedling,
Immunocompetence and survival rate are improved, while Chinese medicine preparation also has stronger luring function, and loach seedling can be promoted the more to take the photograph
Food, builds up health；
Compounding：Weigh 40-50 parts of fermentation fish meals, 10-15 parts of Meretrix meretrix Glycopeptides, 20-30 parts of fish oil micro-capsules, 15-20 parts of Chinese medicine systems
Agent, 20-22 portions of salt, 5-6 parts of Yolk antibodies, 2-4 parts of sodium butyrates, 5-6 parts of carrotene, 2-3 parts of sulfa powders are simultaneously uniformly mixed,
40-60 mesh is smashed it through to sieve up to loach seedling nutrition enhancer；Reinforcing agent has stronger food calling smell, can attract loach
Seedling is gathered in around reinforcing agent and feed, is increased seedling and is ingested, reinforcing agent also has stronger antioxidation, can remove
Extra free radical in loach seedling body, while reinforcing agent also has bacteriostasis antibiosis activity, can effectively enhance loach seedling
Immunocompetence enhances loach seedling constitution and vigor, improves eyesight, the anti-stress ability of loach seedling, improves raising speed, drop
Low aquaculture cost.
Preferably, it is 1 that mixed bacteria, which is weight ratio,:The mixing of the lactobacillus acidophilus, bacillus subtilis of 1.5-1.8
Bacterium, lactobacillus acidophilus, bacillus subtilis bacterium activity be 4000-5000u/g；Bacillus subtilis makes the yield of protease
Rise, more high molecular weight proteins are converted into the peptide of small molecule in fish meal, show as the raising of small peptide content；But bacillus subtilis
Bacterium produces a large amount of metabolite during fermentation, while also increases the content of Volatile Base Nitrogen；Add in acidophilus
In the case of lactobacillus, although the opposite excessive decomposition for reducing, avoiding protein in fish meal of small peptide content, meanwhile, acidophilus
The lactic acid that lactobacillus generates can also inhibit the growth of miscellaneous bacteria and reduce the generation of Volatile Base Nitrogen to a certain extent；Meet
Requirements of the GB/T19164-2003 to fish meal TVB-N content.
Preferably, the mass fraction of Arabic gum aqueous solution is 18-20%, the mass fraction of chitosan solution is 1.0-
1.1%, it dissolves the chitosan in the aqueous acetic acid of 1.0-2.0% volume fractions up to chitosan solution；Chitosan-I
The effect that primary glue agglomerates fish oil again is preferable, and embedding rate and wall material utilization rate are higher.
Compared with prior art, the advantage of the invention is that：1）With n-butanol hydrogen can occur for 1,3,5- trimethoxy-benzene
The effects that key connection, and then the extraction efficiency of system is greatly enhanced, improve the purity and yield of Meretrix meretrix Glycopeptide, gained Meretrix meretrix Glycopeptide
Purity can reach 97%, sugar and protein content each 54% and 42%, Meretrix meretrix Glycopeptide have excellent antitumor activity, suppression
Bacterium antibacterial activity can effectively enhance the immunocompetence of loach seedling, enhance loach seedling constitution and vigor；2）Reinforcing agent has
Stronger food calling smell, can attract loach seedling to be gathered in around reinforcing agent and feed, increase seedling and ingest, reinforcing agent also has
There is stronger antioxidation, free radical extra in loach seedling body can be removed, while reinforcing agent also has bacteriostasis antibiosis
Activity can effectively enhance the immunocompetence of loach seedling, enhance loach seedling constitution and vigor, improve regarding for loach seedling
Power, anti-stress ability improve raising speed, reduce aquaculture cost.
Specific embodiment
The present invention program is described further below by embodiment：
Embodiment 1：
The preparation method of loach seedling nutrition enhancer, includes the following steps：
1）Take fish meal with distilled water according to solid-liquid ratio 1:2 ratio is uniformly mixed, and is inoculated with the mixed bacteria into 3.5%, in 32 DEG C of temperature
The lower fermentation time of degree 12 hours, low temperature drying after fermentation obtains fermentation fish meal；2）Clam is taken through spitting sand, decladding, freeze-drying, crushing
Clam dry powder is obtained afterwards, by solid-liquid ratio 1:6 are dissolved in distilled water and are homogenized, and centrifuge 30-45 minutes, collect in 5 DEG C, 500r/min
Supernatant, then through Hollow Fiber Ultrafiltration column ultrafiltration, ultrafiltration column molecular cut off is 8 × 104, ultrafiltration collection small-molecular-weight ultrafiltrate,
The chloroform of 1/5 volume, the n-butanol of 1/25 volume are added in ultrafiltrate, adds 1,3,5- trimethoxies of n-butanol weight 2%
Base benzene extracts 2 times；Extract liquor is dialysed, is centrifuged, be dried in vacuo after obtain Meretrix meretrix Glycopeptide crude product, after crude product is water-soluble in centrifuging and taking
Clearly, then with 0.2mol/L, 0.4mol/L, 0.6mol/L, 0.8mol/L, 1.0mol/L sodium chloride solution gradient elution, clam is obtained
Glycopeptide；3）0.46 part of tween is added in 100 parts by weight gumwaters to be uniformly mixed, and adds in 12 parts of fish oil, interval ultrasound 5
Minute, supersonic frequency 25kHz, ultrasonic density are 0.35W/cm2, second mode is ultrasound 2 seconds, pause 4 seconds, forms uniform breast
Shape liquid；300 parts of chitosan solutions are added drop-wise to dropwise in the Arabic gum fish oil emulsion being stirred continuously, with hydrochloric acid and hydroxide
Sodium solution adjustment pH is 4.5, is stirred 30 minutes at 18 DEG C；Stratification, condensation product enrichment phase are chitosan-Arabic gum
Fish oil micro-capsule is agglomerated again；4）Weigh 10 parts of radix scutellariaes, 3 portions of Chinese galls, 3 parts of Enteromorphas, 10 portions of cloves, 2 portions of radix polygonati officinalis, 5 parts of Radix Astragalis, 10
Part Herba Epimedii smashes it through 120 mesh sieve, traditional Chinese medicine powder is taken to add water to cook 60 minutes, supernatant concentration is taken after natural cooling to ointment
Up to Chinese medicine preparation；5）Weigh 40 parts of fermentation fish meals, 10 parts of Meretrix meretrix Glycopeptides, 20 parts of fish oil micro-capsules, 15 parts of Chinese medicine preparations, 20 parts
Salt, 5 parts of Yolk antibodies, 2 parts of sodium butyrates, 5 parts of carrotene, 2 parts of sulfa powders are simultaneously uniformly mixed, and are smashed it through 40 mesh and are sieved to obtain the final product
Loach seedling nutrition enhancer.
Embodiment 2：
The preparation method of loach seedling nutrition enhancer, including：Fish meal fermentation, the preparation of Meretrix meretrix Glycopeptide, fish oil micro-capsule system
Standby, Chinese medicine preparation preparation, compounding, specifically include following steps：
1）Fish meal ferments：Take fish meal with distilled water according to solid-liquid ratio 1:3 ratio is uniformly mixed, and is inoculated with the Mixed Microbes into 4.5%
Kind, mixed bacteria is that weight ratio is 1:1.5 lactobacillus acidophilus, the Mixed Microbes of bacillus subtilis, lactobacillus acidophilus, withered grass bud
The bacterium activity of spore bacillus is 5000u/g, and fermentation time 16 hours at a temperature of 33 DEG C, after fermentation low temperature drying must ferment
Fish meal；Small peptide content of fermentation fish meal reaches 29.8% under the conditions of this, and Volatile Base Nitrogen is only 40.3mg/100g, restricted amino
The content of acid promotes more than 30.2%, and the more common fish meal of total antioxidant capacity promotes more than 70%, has excessive in removing organism
The great potential of free radical；In addition, in fermentation fish meal, the limiting amino acids such as lysine, threonine, methionine, histidine
Content is all obviously improved, and disclosure satisfy that demand of the loach seedling to limiting amino acid；
2）The preparation of Meretrix meretrix Glycopeptide：Clam is taken to obtain clam dry powder after spitting sand, decladding, freeze-drying, crushing, by solid-liquid ratio 1:7 is molten
It in distilled water and is homogenized, is centrifuged 45 minutes in 6 DEG C, 600r/min, collect supernatant, then through Hollow Fiber Ultrafiltration column ultrafiltration, surpass
Filter column molecular cut off is 8 × 104, ultrafiltration collect small-molecular-weight ultrafiltrate, added in ultrafiltrate 2/5 volume chloroform, 1/
The n-butanol of 20 volumes adds 1,3,5- trimethoxy-benzenes of n-butanol weight 6%, extracts 3 times；Extract liquor is dialysed, from
Obtain Meretrix meretrix Glycopeptide crude product after the heart, vacuum drying, centrifuging and taking supernatant after crude product is water-soluble, then with 0.2mol/L, 0.4mol/L,
0.6mol/L, 0.8mol/L, 1.0mol/L sodium chloride solution gradient elution, obtain Meretrix meretrix Glycopeptide；1,3,5- trimethoxy-benzene can
The effects that hydrogen bond is connect occurs, and then greatly enhance the extraction efficiency of system with n-butanol, improve the purity of Meretrix meretrix Glycopeptide with obtaining
Rate, the purity of gained Meretrix meretrix Glycopeptide can reach 97%, and sugar and protein content each 54% and 42%, Meretrix meretrix Glycopeptide have excellent
Antitumor activity, bacteriostasis antibiosis activity, can effectively enhance the immunocompetence of loach seedling, enhance loach seedling constitution and work
Power；
3）The preparation of fish oil micro-capsule：0.56 part of Tween-80 is added in 120 parts by weight gumwaters to be uniformly mixed, and is added
Enter 14 parts of fish oil, interval ultrasound 6 minutes, supersonic frequency 30kHz, ultrasonic density are 0.45W/cm2, second mode for ultrasound 2 seconds,
Pause 4 seconds, forms uniform emulsion；400 parts of chitosan solutions are added drop-wise to the Arabic gum fish oil breast being stirred continuously dropwise
Change in liquid, be 5.0 with hydrochloric acid and sodium hydroxide solution adjustment pH, stirred 45 minutes at 20 DEG C；Stratification, condensation product enrichment
It is mutually that chitosan-Arabic gum agglomerates fish oil micro-capsule again；Rich in omega-3 polyunsaturated fatty acids EPA and DHA, tool in fish oil
Having reduces serum cholesterol and low-density lipoprotein content, improves the different physiological roles such as eyesight, anticoagulation, anticancer, also has
Improve the effect of organism lipid structure；But it is extremely sensitive to light, heat and oxygen etc. meanwhile containing multiple double bonds in EPA and DHA,
It needs to be protected from light, low temperature and sealing storage and transportation, largely limits application of the fish oil in industrial circle；Chitosan-Arab
Glue agglomerates fish oil micro-capsule in regular spherical shape again, and favorable dispersibility, particle diameter distribution is uniform, and average grain diameter is 16-20 μm, right
The embedding rate of fish oil and the utilization rate of wall material are respectively 75.33% and 81.01%, can be preferable hidden by fish oil, drop
The influence of low its light, heat, oxygen improves fish oil utilization rate；
4）The preparation of Chinese medicine preparation：Weigh 12 parts of radix scutellariaes, 4 portions of Chinese galls, 4 parts of Enteromorphas, 15 portions of cloves, 5 portions of radix polygonati officinalis, 8 parts of Radix Astragalis,
12 parts of Herba Epimedii smash it through 160 mesh sieve, traditional Chinese medicine powder are taken to add water to cook 90 minutes, supernatant concentration is taken after natural cooling to soft
Cream is up to Chinese medicine preparation；Chinese medicine preparation safety, low toxicity, low-residual, are not likely to produce drug resistance, and tonifying spleen, stomach invigorating, tonifying Qi, solid table are adjusted
Section metabolism, enhances spleen and stomach function, can reduce stress reaction, the incidence of loach seedling, improves immunocompetence and survival rate, together
When Chinese medicine preparation also have stronger luring function, loach seedling can be promoted the more to ingest, built up health；
5）Compounding：Weigh 50 parts of fermentation fish meals, 15 parts of Meretrix meretrix Glycopeptides, 30 parts of fish oil micro-capsules, 20 parts of Chinese medicine preparations, 22 portions of foods
Salt, 6 parts of Yolk antibodies, 4 parts of sodium butyrates, 6 parts of carrotene, 3 parts of sulfa powders are simultaneously uniformly mixed, and are smashed it through 60 mesh and are sieved up to mud
Loach seedling nutrition enhancer；Reinforcing agent has stronger food calling smell, and loach seedling can be attracted to be gathered in reinforcing agent and feed
Around, increase seedling and ingest, reinforcing agent also has stronger antioxidation, can remove freedom extra in loach seedling body
Base, while reinforcing agent also has bacteriostasis antibiosis activity, can effectively enhance the immunocompetence of loach seedling, enhance loach seedling body
Matter and vigor improve eyesight, the anti-stress ability of loach seedling, improve raising speed, reduce aquaculture cost.
The mass fraction of Arabic gum aqueous solution is 20%, and the mass fraction of chitosan solution is 1.1%, and chitosan is molten
Solution is in the aqueous acetic acid of 2.0% volume fraction up to chitosan solution；Chitosan-Arabic gum agglomerates the effect of fish oil again
Preferably, embedding rate and wall material utilization rate are higher.
Embodiment 3：
The preparation method of loach seedling nutrition enhancer, including：Fish meal fermentation, the preparation of Meretrix meretrix Glycopeptide, fish oil micro-capsule system
Standby, Chinese medicine preparation preparation, compounding, specifically include following steps：
Fish meal ferments：Take fish meal with distilled water according to solid-liquid ratio 1:2.5 ratio is uniformly mixed, and is inoculated with the Mixed Microbes into 4.2%
Kind, fermentation time 12 hours at a temperature of 32 DEG C, after fermentation low temperature drying obtains fermentation fish meal；Fermentation fish meal is small under the conditions of this
Peptide content reaches 29.8%, and Volatile Base Nitrogen is only 40.3mg/100g, and the content of limiting amino acid promotes more than 30.2%,
The more common fish meal of total antioxidant capacity promotes more than 70%, has the great potential for removing excessive free radicals in organism；In addition,
In fermentation fish meal, the content of the limiting amino acids such as lysine, threonine, methionine, histidine is all obviously improved, can
Meets the needs of loach seedling is to limiting amino acid；
The preparation of Meretrix meretrix Glycopeptide：Clam is taken to obtain clam dry powder after spitting sand, decladding, freeze-drying, crushing, by solid-liquid ratio 1:It is 6.5 molten
It in distilled water and is homogenized, is centrifuged 35 minutes in 5 DEG C, 560r/min, collect supernatant, then through Hollow Fiber Ultrafiltration column ultrafiltration, surpass
Filter column molecular cut off is 8 × 104, ultrafiltration collect small-molecular-weight ultrafiltrate, added in ultrafiltrate 1/5 volume chloroform, 1/
The n-butanol of 22 volumes adds 1,3,5- trimethoxy-benzenes of n-butanol weight 3%, extracts 3 times；Extract liquor is dialysed, from
Obtain Meretrix meretrix Glycopeptide crude product after the heart, vacuum drying, centrifuging and taking supernatant after crude product is water-soluble, then with 0.2mol/L, 0.4mol/L,
0.6mol/L, 0.8mol/L, 1.0mol/L sodium chloride solution gradient elution, obtain Meretrix meretrix Glycopeptide；1,3,5- trimethoxy-benzene can
The effects that hydrogen bond is connect occurs, and then greatly enhance the extraction efficiency of system with n-butanol, improve the purity of Meretrix meretrix Glycopeptide with obtaining
Rate, the purity of gained Meretrix meretrix Glycopeptide can reach 97%, and sugar and protein content each 54% and 42%, Meretrix meretrix Glycopeptide have excellent
Antitumor activity, bacteriostasis antibiosis activity, can effectively enhance the immunocompetence of loach seedling, enhance loach seedling constitution and work
Power；
The preparation of fish oil micro-capsule：0.55 part of Tween-80 is added in 110 parts by weight gumwaters to be uniformly mixed, and is added in
12 parts of fish oil, interval ultrasound 6 minutes, supersonic frequency 28kHz, ultrasonic density are 0.38W/cm2, second mode is 2 seconds ultrasonic, temporary
Stop 4 seconds, form uniform emulsion；350 parts of chitosan solutions are added drop-wise to the Arabic gum fish oil emulsification being stirred continuously dropwise
In liquid, it is 4.6 with hydrochloric acid and sodium hydroxide solution adjustment pH, is stirred 35 minutes at 18 DEG C；Stratification, condensation product enrichment phase
As chitosan-Arabic gum agglomerates fish oil micro-capsule again；Rich in omega-3 polyunsaturated fatty acids EPA and DHA in fish oil, have
It reduces serum cholesterol and low-density lipoprotein content, improve the different physiological roles such as eyesight, anticoagulation, anticancer, also have and change
The effect of kind organism lipid structure；But it is extremely sensitive to light, heat and oxygen etc. meanwhile containing multiple double bonds in EPA and DHA, it needs
Be protected from light, low temperature and sealing storage and transportation, largely limit application of the fish oil in industrial circle；Chitosan-Arabic gum
Cohesion fish oil micro-capsule is in regular spherical shape again, and favorable dispersibility, particle diameter distribution is uniform, and average grain diameter is 16-20 μm, to fish
The embedding rate of oil and the utilization rate of wall material are respectively 75.33% and 81.01%, can be preferable hidden by fish oil, are reduced
The influence of its light, heat, oxygen improves fish oil utilization rate；
The preparation of Chinese medicine preparation：Weigh 10 parts of radix scutellariaes, 3 portions of Chinese galls, 3 parts of Enteromorphas, 12 portions of cloves, 4 portions of radix polygonati officinalis, 6 parts of Radix Astragalis, 10
Part Herba Epimedii smashes it through 140 mesh sieve, traditional Chinese medicine powder is taken to add water to cook 80 minutes, supernatant concentration is taken after natural cooling to ointment
Up to Chinese medicine preparation；Chinese medicine preparation safety, low toxicity, low-residual, are not likely to produce drug resistance, and tonifying spleen, stomach invigorating, tonifying Qi, solid table are adjusted
Metabolism enhances spleen and stomach function, can reduce stress reaction, the incidence of loach seedling, improves immunocompetence and survival rate, simultaneously
Chinese medicine preparation also has stronger luring function, and loach seedling can be promoted the more to ingest, built up health；
Compounding：Weigh 45 parts of fermentation fish meals, 14 parts of Meretrix meretrix Glycopeptides, 28 parts of fish oil micro-capsules, 16 parts of Chinese medicine preparations, 20 parts of salt, 5
Part Yolk antibody, 3 parts of sodium butyrates, 6 parts of carrotene, 3 parts of sulfa powders are simultaneously uniformly mixed, and are smashed it through 60 mesh and are sieved up to loach children
Seedling nutrition enhancer；Reinforcing agent has stronger food calling smell, and loach seedling can be attracted to be gathered in around reinforcing agent and feed,
Increase seedling to ingest, reinforcing agent also has stronger antioxidation, can remove free radical extra in loach seedling body, together
When reinforcing agent also there is bacteriostasis antibiosis activity, can effectively enhance the immunocompetence of loach seedling, enhancing loach seedling constitution with
Vigor improves eyesight, the anti-stress ability of loach seedling, improves raising speed, reduces aquaculture cost.
Mixed bacteria is that weight ratio is 1:1.6 lactobacillus acidophilus, the Mixed Microbes of bacillus subtilis, lactobacillus acidophilus,
The bacterium activity of bacillus subtilis is 4500u/g；Bacillus subtilis rises the yield of protease, more big in fish meal
Molecule protein is converted into the peptide of small molecule, shows as the raising of small peptide content；But bacillus subtilis produces during fermentation
A large amount of metabolite has been given birth to, while has also increased the content of Volatile Base Nitrogen；In the case of adding in lactobacillus acidophilus, although
Small peptide content is opposite to be reduced, but avoid the excessive decomposition of protein in fish meal, meanwhile, the lactic acid that lactobacillus acidophilus generates also can
Inhibit the growth of miscellaneous bacteria to a certain extent and reduce the generation of Volatile Base Nitrogen；Meet GB/T19164-2003 to wave fish meal
The requirement of hair property alkali nitrogen content.
The mass fraction of Arabic gum aqueous solution is 20%, and the mass fraction of chitosan solution is 1.0%, and chitosan is molten
Solution is in the aqueous acetic acid of 1.0% volume fraction up to chitosan solution；Chitosan-Arabic gum agglomerates the effect of fish oil again
Preferably, embedding rate and wall material utilization rate are higher.
Routine operation in operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme of the present invention is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (9)
1. the preparation method of loach seedling nutrition enhancer, including：Fish meal fermentation, the preparation of Meretrix meretrix Glycopeptide, fish oil micro-capsule
It prepares, the preparation of Chinese medicine preparation, compound, it is characterised in that：The preparation process of the Meretrix meretrix Glycopeptide is：Clam is taken through spitting sand, going
Clam dry powder is obtained after shell, freeze-drying, crushing, by solid-liquid ratio 1:6-7 is dissolved in distilled water and is homogenized, in 5-6 DEG C, 500-600r/
Min is centrifuged 30-45 minutes, collects supernatant, then through Hollow Fiber Ultrafiltration column ultrafiltration, ultrafiltration column molecular cut off is 8 × 104, surpass
Small-molecular-weight ultrafiltrate is collected in filter, and the chloroform of 1/5-2/5 volumes, the n-butanol of 1/25-1/20 volumes are added in ultrafiltrate, then
1,3, the 5- trimethoxy-benzenes of n-butanol weight 2-6% are added in, are extracted 2-3 times；Extract liquor is dialysed, is centrifuged, be dried in vacuo after
To Meretrix meretrix Glycopeptide crude product, centrifuging and taking supernatant after crude product is water-soluble, then with 0.2mol/L, 0.4mol/L, 0.6mol/L, 0.8mol/L,
1.0mol/L sodium chloride solution gradient elutions, obtain Meretrix meretrix Glycopeptide.
2. the preparation method of loach seedling nutrition enhancer according to claim 1, it is characterised in that：The Meretrix meretrix Glycopeptide
Preparation process in chloroform additive amount be ultrafiltrate volume 1/5-2/5, the additive amount of n-butanol is the 1/ of ultrafiltrate volume
, the additive amount of 1,3,5- trimethoxy-benzene is the 2-6% of n-butanol weight.
3. the preparation method of loach seedling nutrition enhancer according to claim 1, it is characterised in that：The fish meal fermentation
Step is：Take fish meal with distilled water according to solid-liquid ratio 1:The ratio of 2-3 is uniformly mixed, and is inoculated with the mixed bacteria into 3.5-4.5%,
Fermentation time 12-16 hours at a temperature of 32-33 DEG C, low temperature drying after fermentation obtains fermentation fish meal.
4. the preparation method of loach seedling nutrition enhancer according to claim 3, it is characterised in that：The fish meal fermentation
Mixed bacteria in step is that weight ratio is 1:The lactobacillus acidophilus of 1.5-1.8, the Mixed Microbes of bacillus subtilis, acidophilus breast bar
Bacterium, bacillus subtilis bacterium activity be 4000-5000u/g.
5. the preparation method of loach seedling nutrition enhancer according to claim 1, it is characterised in that：The micro- glue of fish oil
The preparation process of capsule is：0.46-0.56 parts of Tween-80s are added in 100-120 parts by weight gumwaters to be uniformly mixed, and are added
Enter 12-14 parts of fish oil, interval ultrasound 5-6 minutes forms uniform emulsion；300-400 parts of chitosan solutions are added dropwise dropwise
It is 4.5-5.0 with hydrochloric acid and sodium hydroxide solution adjustment pH, in 18-20 into the Arabic gum fish oil emulsion being stirred continuously
It is stirred 30-45 minutes at DEG C；Stratification, condensation product enrichment phase are that chitosan-Arabic gum agglomerates fish oil micro-capsule again.
6. the preparation method of loach seedling nutrition enhancer according to claim 5, it is characterised in that：The micro- glue of fish oil
The mass fraction of Arabic gum aqueous solution in the preparation process of capsule is 18-20%, and the mass fraction of chitosan solution is 1.0-
1.1%, it dissolves the chitosan in the aqueous acetic acid of 1.0-2.0% volume fractions up to chitosan solution.
7. the preparation method of loach seedling nutrition enhancer according to claim 5, it is characterised in that：The micro- glue of fish oil
The supersonic frequency of interval ultrasound in the preparation process of capsule is 25-30kHz, and ultrasonic density is 0.35-0.45W/cm2, second mode
For ultrasound 2 seconds, pause 4 seconds.
8. the preparation method of loach seedling nutrition enhancer according to claim 1, it is characterised in that：The Chinese medicine preparation
Preparation process be：Weigh 10-12 parts of radix scutellariaes, 3-4 portions of Chinese galls, 3-4 parts of Enteromorphas, 10-15 portions of cloves, 2-5 parts of radix polygonati officinalis, 5-8
Part Radix Astragali, 10-12 parts of Herba Epimedii smash it through 120-160 mesh sieve, traditional Chinese medicine powder are taken to add water to cook 60-90 minutes, after natural cooling
Take supernatant concentration to ointment up to Chinese medicine preparation.
9. the preparation method of loach seedling nutrition enhancer according to claim 1, it is characterised in that：The compounding step
For：Weigh 40-50 parts of fermentation fish meals, 10-15 parts of Meretrix meretrix Glycopeptides, 20-30 parts of fish oil micro-capsules, 15-20 parts of Chinese medicine preparations, 20-
22 portions of salt, 5-6 parts of Yolk antibodies, 2-4 parts of sodium butyrates, 5-6 parts of carrotene, 2-3 parts of sulfa powders are simultaneously uniformly mixed, after crushing
40-60 mesh is crossed to sieve up to loach seedling nutrition enhancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711238844.XA CN108185213A (en) | 2017-11-30 | 2017-11-30 | The preparation method of loach seedling nutrition enhancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711238844.XA CN108185213A (en) | 2017-11-30 | 2017-11-30 | The preparation method of loach seedling nutrition enhancer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108185213A true CN108185213A (en) | 2018-06-22 |
Family
ID=62573512
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711238844.XA Withdrawn CN108185213A (en) | 2017-11-30 | 2017-11-30 | The preparation method of loach seedling nutrition enhancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108185213A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108821412A (en) * | 2018-07-02 | 2018-11-16 | 河南牧业经济学院 | A kind of improver of water quality and preparation method thereof for aquaculture |
| CN108911161A (en) * | 2018-07-02 | 2018-11-30 | 河南牧业经济学院 | A kind of water conditioner and preparation method thereof for aquaculture |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103549194A (en) * | 2013-10-31 | 2014-02-05 | 白国丰 | Special Chinese herbal medicine phagostimulant for loaches |
| CN103750047A (en) * | 2014-01-16 | 2014-04-30 | 安徽天邦饲料科技有限公司 | Non-antibiotic micropowder aged compound hard particle feed for loaches and preparation method thereof |
| CN104222640A (en) * | 2014-09-15 | 2014-12-24 | 天津农学院 | Special feed for loaches |
| CN105343415A (en) * | 2015-11-03 | 2016-02-24 | 安徽孔津湖农业开发有限公司 | Method for preventing diseases of loaches |
| CN105495057A (en) * | 2015-12-27 | 2016-04-20 | 天津现代天骄水产饲料股份有限公司 | Feed special for young loaches |
-
2017
- 2017-11-30 CN CN201711238844.XA patent/CN108185213A/en not_active Withdrawn
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103549194A (en) * | 2013-10-31 | 2014-02-05 | 白国丰 | Special Chinese herbal medicine phagostimulant for loaches |
| CN103750047A (en) * | 2014-01-16 | 2014-04-30 | 安徽天邦饲料科技有限公司 | Non-antibiotic micropowder aged compound hard particle feed for loaches and preparation method thereof |
| CN104222640A (en) * | 2014-09-15 | 2014-12-24 | 天津农学院 | Special feed for loaches |
| CN105343415A (en) * | 2015-11-03 | 2016-02-24 | 安徽孔津湖农业开发有限公司 | Method for preventing diseases of loaches |
| CN105495057A (en) * | 2015-12-27 | 2016-04-20 | 天津现代天骄水产饲料股份有限公司 | Feed special for young loaches |
Non-Patent Citations (3)
| Title |
|---|
| 刘恺等: "壳聚糖/阿拉伯胶复凝聚鱼油微胶囊的制备及性质研究", 《功能材料》 * |
| 吴杰连等: "文蛤糖肽MGP0501的分离纯化及性质鉴定", 《氨基酸和生物资源》 * |
| 林俊宏等: "混合菌固态发酵制备发酵鱼粉的研究", 《饲料工业》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108821412A (en) * | 2018-07-02 | 2018-11-16 | 河南牧业经济学院 | A kind of improver of water quality and preparation method thereof for aquaculture |
| CN108911161A (en) * | 2018-07-02 | 2018-11-30 | 河南牧业经济学院 | A kind of water conditioner and preparation method thereof for aquaculture |
| CN108911161B (en) * | 2018-07-02 | 2021-04-16 | 河南牧业经济学院 | Water quality regulator for aquaculture and preparation method thereof |
| CN108821412B (en) * | 2018-07-02 | 2021-04-30 | 河南牧业经济学院 | Water quality improver for aquaculture and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102940214B (en) | Fruit and vegetable fiber nutrition powder production and production method and application thereof | |
| TWI548356B (en) | Probiotics-containing soybean oligosaccharide product and preparation thereof | |
| CN103340300A (en) | Preparation method for green ecological crucian forage | |
| CN106174474A (en) | A kind of it is applicable to the special dietary seafood that Patients with Cardiovascular/Cerebrovascular Diseases is edible | |
| CN108813634A (en) | A kind of fast food type yolk oligopeptides-phosphatide type DHA economic benefits and social benefits dietary supplements and preparation method thereof | |
| CN102357212B (en) | Compound product obtained through supercritical CO2 extraction, and preparation method thereof | |
| CN105768106A (en) | Liver nutrition improving functional food | |
| CN109699745A (en) | A kind of fucosylated oligosaccharide baby formula milk powder and preparation method thereof | |
| KR102339573B1 (en) | Functional Feed Additive Composition For pets | |
| CN108185213A (en) | The preparation method of loach seedling nutrition enhancer | |
| JP2002065206A (en) | Immunostimulating food | |
| KR102564700B1 (en) | Complex functional protein shake and preparation method thereof | |
| CN113186066A (en) | SOD wine capable of preserving health and tonifying kidney and preparation method thereof | |
| WO2019100540A1 (en) | Method for preparing litopenaeusvannamei feed additive | |
| JP2002209552A (en) | Immunostimulating food | |
| CN105558260B (en) | Potato peptide nutrient food and preparation method thereof | |
| JP2001224330A (en) | Food comprising lactobacillus symbiotic culture product and medicinal plant and method for producing the same | |
| CN112352887B (en) | Compound feed for mandarin fish and preparation method thereof | |
| CN105154304A (en) | Black garlic vinegar and preparing method thereof | |
| CN108208311A (en) | Boar feedstuff preparation method containing sea cucumber active peptides | |
| JP2008255057A (en) | Anticancer agent, method for producing anticancer agent and food and drink | |
| JP5034071B2 (en) | Kawariharatake extract that suppresses breast cancer | |
| CN113068784A (en) | SOD beverage for relieving body pain and preparation method thereof | |
| CN106260497A (en) | A kind of giant salamander nourishing egg albumen powder production technology | |
| CN106036873A (en) | Hami melon-flavor mint cool wolfberry red date chewable tablet and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20180622 |