CN105639204B - A kind of biology toner and its application in aquaculture of aquatic animal - Google Patents
A kind of biology toner and its application in aquaculture of aquatic animal Download PDFInfo
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- CN105639204B CN105639204B CN201610031287.3A CN201610031287A CN105639204B CN 105639204 B CN105639204 B CN 105639204B CN 201610031287 A CN201610031287 A CN 201610031287A CN 105639204 B CN105639204 B CN 105639204B
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Abstract
The invention discloses a kind of biological toner, raw material compositions are as follows: 60-80 parts of red phaffia rhodozyma spray powder, 8-12 parts of red phaffia rhodozyma freeze-dried powder, 8-12 parts of lactobacillus-fermented product, 4-8 parts of antibacterial peptide, 2-4 parts of mountain flour, 0.5-1.5 parts of glutamine.Application the invention also discloses the preparation method of the biology toner and in aquaculture of aquatic animal.For biological toner of the invention using red phaffia rhodozyma as effective component, nutritive value is abundant, and protein content is high and is rich in various trace elements and coenzyme, also it is rich in astaxanthin, and a variety of nutriments are equipped with, as aquaculture of aquatic animal toner, have the advantages that hyperchromic effect stability.
Description
Technical field
The present invention relates to a kind of biological toner and its applications in aquaculture of aquatic animal, belong to aquaculture of aquatic animal skill
Art field.
Background technique
With the fast development of artificial breeding industry, the transition of environment, the change of food chain makes the body colour of aquiculture animal
Different degrees of change has occurred.The fishes and shrimps grown in its natural state are bright in luster, delicious meat, and support in artificial intensive
Under the conditions of growing, due to largely using artifical compound feed, the culture-cycle shortening of fish, shrimp, crab can only obtain a small amount of Natural Water
Biology, and effective color source is few in feed and ingredient is unstable, causes the body colour of fish, shrimp, crab thin out, channel catfish " becomes
It is white ", " become flower " even " blackening "；Yellow wood fish " bleaching ", " blackening "；Finless eel and mud " rubescent "；Bream becomes " goldfish "；Carp, crucian carp
" blackening ", " reddening "；Black carp " whitens "；Grass carp " golden yellow " etc..These abnormal body colours reduce the commodity value of cultured fishes,
Economic loss is brought to culturist.Therefore, the body colour for how improving Aquatic farming animals is allowed to the body colour phase with wild fishes and shrimps
It is close, it is always research important in aquaculture field and practices project.
The spot billows color that aquatic livestock can show, is mainly determined by carotenoid, also with melanin, guanine etc.
Pigment group is related, but aquatic livestock itself cannot synthesize carotenoid, thus carotenoid must be absorbed from food.
Colorant is by sources generally divided into two kinds, i.e. natural colorant and synthetic coloring matter.Natural colorant refers to some rich in class Hu
Yellow, red, purple the animal such as radish element and lutein and its extract, such as oppossum shrimp, squama shrimp；Plant and its extract, it is such as beautiful
Rice, carrot, lucerne potato powder, orange peel etc.；Microorganism and its extract, as in yeast, photosynthetic bacteria red spirillum, in miniature algae
Du Shi algae etc.；Spirulina and its extract.Under modern intensive cultivating condition, aquatic livestock ingests natural bait (rich in class
Carrotene) chance it is fewer and fewer, therefore, in feed add carotenoid as colorant, to cultured fishes body colour
Regulated and controled, has caused the extensive attention of research worker.
Summary of the invention
For the above-mentioned prior art, the object of the present invention is to provide a kind of biological toner and its in aquaculture of aquatic animal
Application.For the biology toner using red phaffia rhodozyma as effective component, nutritive value is abundant, and protein content is high and rich in a variety of micro-
Secondary element and coenzyme are also rich in astaxanthin, and are equipped with a variety of nutriments, as aquaculture of aquatic animal toner, have and increase
The advantages of color effect stability.
To achieve the above object, the present invention adopts the following technical solutions:
A kind of biology toner, raw material composition are as follows:
60-80 parts of red phaffia rhodozyma spray powder, 8-12 parts of red phaffia rhodozyma freeze-dried powder, 8-12 parts of lactobacillus-fermented product,
4-8 parts of antibacterial peptide, 2-4 parts of mountain flour, 0.5-1.5 parts of glutamine.
Preferably, the raw material composition of the biology toner are as follows:
70 parts of red phaffia rhodozyma spray powder, 10 parts of red phaffia rhodozyma freeze-dried powder, 10 parts of lactobacillus-fermented product, antibacterial peptide 6
Part, 3 parts of mountain flour, 1 part of glutamine.
Preferably, the red phaffia rhodozyma spray powder and red phaffia rhodozyma freeze-dried powder are to be trained by red phaffia rhodozyma in yeast
It supports progress fermented and cultured on base and obtains fermentation liquid, fermentation liquid is prepared through spray drying and freeze-drying respectively.
The red phaffia rhodozyma is purchased from Chinese industrial Microbiological Culture Collection administrative center, and strain number is
CICC33064, the yeast strain with high-yield astaxanthin function.
The raw material of the yeast culture medium forms are as follows: glucose 1.5%, peptone 1.0%, yeast extract 0.5%, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.15%, magnesium sulfate 0.02%, sodium chloride 2.0%, calcium chloride 0.02%, glutamic acid 0.06%, are mass percent.
Screening is optimized to the composition of yeast culture medium in the present invention, finds after many experiments compare, using the ferment of above-mentioned composition
Female culture medium, the amount and biomass of red Astaxanthin Production by Phaffia rhodozyma are relatively high, and also reduce production cost.
Preferably, the lactobacillus-fermented product is Pediococcus pentosaceus PP (Pediococcus pentosaceus PP) warp
The tunning that fermented and cultured obtains；Pediococcus pentosaceus PP was preserved in China typical culture collection on February 22nd, 2012
Center, deposit number are CCTCC NO:M 2012029；It is documented in an a kind of patent of invention " raising animal life of applicant
In the lactic acid bacteria semisolid fermentation product and technique of long performance " (CN 102599348B).Above-mentioned Pediococcus pentosaceus is a kind of important
Beneficial bacterium, to body have important physiological action.Lactic acid, bacteriocin etc. can be generated during its growth metabolism, reduced
Intestinal pH, to inhibit the growth and breeding of pathogenic bacteria；Intestinal epithelial cell can also be attached to simultaneously, is had with pathogenic microorganism contention
The nutriment of limit and ecological site, play occupancy protective effect etc..The bacterial strain overcomes external environment tolerance difference
Drawback can pass through the low pH in alimentary canal as effective probiotics and there is stronger colonization ability to play its biology
Learn function.Above-mentioned Pediococcus pentosaceus is applied in biological toner by the present invention, can improve the internal Tiny ecosystem of aquatic livestock
Environment, maintenance alimentary canal health.
The present invention also provides the preparation method of the biology toner, steps are as follows: by each component be uniformly mixed to get.
The biology toner is in aquaculture of aquatic animal as the hyperchromic application with feed addictive and guarantor of the invention
Protect range.
In above-mentioned application, specific application method are as follows: add the biology toner in the ratio that mass fraction is 1-3 ‰
Into aquatic animal feed, feeds the period for one or feed for a long time within 30 days.
The present invention also provides a kind of aquaculture of aquatic animal it is hyperchromic use feed, be by aquatic livestock basal feed by quality
Score adds above-mentioned biological toner for 1-3 ‰ and is prepared.
Beneficial effects of the present invention:
(1) biological toner of the invention is using red phaffia rhodozyma as effective component, and nutritive value is abundant, protein content it is high and
Rich in various trace elements and coenzyme, it is also rich in astaxanthin, and be equipped with a variety of nutriments, can be used as aquaculture of aquatic animal use
Toner has the advantages that hyperchromic effect stability.
(2) in biological toner of the invention, red phaffia rhodozyma is added in the form of freeze-dried powder and two kinds of spray powder, wherein
Viable count in freeze-dried powder is high, but preparation cost is also high；Viable count is relatively low in spray powder, but meets nutritional need simultaneously
Preparation cost is low.Comprehensively consider the viable count and production cost in biological toner, the present invention to red phaffia rhodozyma freeze-dried powder and
The additional amount of spray powder is optimized, and makes to have obtained between the hyperchromic effect of biological toner and preparation cost flat well
Weighing apparatus.
(3) in biological toner of the invention, the type of red phaffia rhodozyma is screened, Chinese work is preferably preserved in
Industry Microbiological Culture Collection administrative center, strain number are the red phaffia rhodozyma of CICC33064, and the ability for producing astaxanthin is significant
Better than other bacterial strains.
(4) astaxanthin being rich in biological toner of the invention can be directly long-pending using pigment is completed by aquatic livestock
Tired process directly plays hyperchromic effect, and on the other hand its nutriment being rich in ensure that the nutritional need of aquatic livestock mentions
The high immunity of aquatic livestock, added antibacterial peptide can also play the role of improving body resistance against diseases, while paddy ammonia
Amide can play the effect of maintenance alimentary canal health, and the angle immune from nutrition and raising is that aquatic livestock creates maintenance body colour
Optimum condition.Biological toner of the invention acts on body color of aquatic animal maintenance in terms of directly or indirectly two, and effect is aobvious
It is lasting to write.
Detailed description of the invention
Scale sector area under Fig. 1: 10x mirror；
Chromatophore under Fig. 2: 40x mirror；
Fig. 3: it feeds after 42d and pigment accumulation occurs in albefaction coloured silk crucian carp fin ray base portion；
Specific embodiment
The present invention is further illustrated in conjunction with the embodiments, it should which explanation, following the description is merely to explain this
Invention, is not defined its content.
Embodiment 1: different yeast strains produce astaxanthin ability and compare the formulation optimization with yeast culture medium
1 materials and methods
Strain is used in 1.1 tests
Rhodotorula glutinis (N), deep rhodotorula glutinis (SN), ocean rhodotorula (H), rhodotorula (R8) are that Bora benefit in Shandong is next biological
The existing saccharomycete of biological study institute, Engineering stock Co., Ltd.
Red phaffia rhodozyma (R5) is purchased from Chinese industrial Microbiological Culture Collection administrative center, and strain number is
CICC33064 has the yeast strain for producing astaxanthin function.
1.2 different yeast strains produce astaxanthin ability and compare
Using the basic components of yeast culture medium as fermentation medium, fermented and cultured is carried out to different yeast strains, is adopted
Its ability for producing astaxanthin is measured with dimethyl sulfoxide method.
The basic components of yeast culture medium: glucose 1.5%, peptone 1.0%, yeast extract 0.5%, potassium dihydrogen phosphate
0.15%, magnesium sulfate 0.01%, sodium chloride 1.0%；It is mass percent.
The formulation optimization of 1.3 yeast culture mediums
Using astaxanthin and biomass as evaluation index, pass through the formula composition of optimization of orthogonal test yeast culture medium.
The measuring method of astaxanthin: dimethyl sulfoxide method；
The measuring method of biomass: after fermentation liquid distilled water is centrifuged (4000r/min, 15min) washing three times, 105
DEG C drying 24 hours, weighing, unit mg/mL.
1 orthogonal design formula composition of table
The screening of 1.4 wall breaking enzymes
Using combined-enzyme method broken wall compared with the astaxanthin that dimethyl sulfoxide method measures, sporoderm-broken rate is determined.
2 broken wall of table enzyme orthogonal design formula
2 test results
The production astaxanthin ability of 2.1 different saccharomycete
3 bacterial strain of table produces the screening of astaxanthin ability
As can be seen from Table 3, strain number is that the production astaxanthin ability of the red phaffia rhodozyma (R5) of CICC33064 is obviously excellent
In other yeast strains.
4 biomass of table and astaxanthin yield change over time trend
As can be seen from Table 4, each saccharomycete is stepped up with the amount of the extension synthesizing astaxanthin of incubation time, but red method
Husband's yeast (R5) aggregate velocity is significantly faster than that other bacterial strains, therefore R5 is ultimately determined to be used to prepare the target of biological toner
Bacterial strain.
The formulation optimization result of 2.2 yeast culture mediums
5 formulation optimization result of table
As shown in Table 5, formula 5 and 7 effects of formula are preferable, but are formulated 5 in cost lower than formula 7, therefore finally determine
The formula of yeast culture medium is formula 5, i.e. glucose 1.5%, peptone 1.0%, yeast extract 0.5%, potassium dihydrogen phosphate
0.15%, magnesium sulfate 0.02%, sodium chloride 2.0%, calcium chloride 0.02%, glutamic acid 0.06%, are mass percent.
The selection result of 2.3 wall breaking enzymes
6 complex enzyme broken wall result of table
As shown in Table 6,5 sporoderm-broken rate is formulated up to 85%, and effect is preferable, determines that it is the prescription of compound wall breaking enzyme.
Embodiment 2: the preparation of biological toner
1 raw material composition
It is made of the raw material of following parts by weight:
70 parts of red phaffia rhodozyma spray powder, 10 parts of red phaffia rhodozyma freeze-dried powder, 10 parts of lactobacillus-fermented product, antibacterial peptide 6
Part, 3 parts of mountain flour, 1 part of glutamine.
Wherein, red phaffia rhodozyma spray powder and red phaffia rhodozyma freeze-dried powder, be by red phaffia rhodozyma in yeast culture medium
It carries out fermented and cultured and obtains fermentation liquid, fermentation liquid is prepared through spray drying and freeze-drying respectively.Fermented and cultured and spray drying, freeze-drying operation
It is conventional technical means in the prior art.
Red phaffia rhodozyma is purchased from Chinese industrial Microbiological Culture Collection administrative center, strain number CICC33064, tool
There is the yeast strain for producing astaxanthin function.
The raw material of yeast culture medium forms are as follows: glucose 1.5%, peptone 1.0%, yeast extract 0.5%, potassium dihydrogen phosphate
0.15%, magnesium sulfate 0.02%, sodium chloride 2.0%, calcium chloride 0.02%, glutamic acid 0.06%, are mass percent.
Lactobacillus-fermented product, which is that Pediococcus pentosaceus PP (Pediococcus pentosaceus PP) is fermented, to be cultivated
The tunning arrived；Pediococcus pentosaceus PP was preserved in China typical culture collection center on February 22nd, 2012, protected
Hiding number is CCTCC NO:M 2012029.
Antibacterial peptide is the product of Shandong Baolai-leelai Bio-engineering Co., Ltd., product name: antibacterial peptide CEC-38.
2 preparation methods
Weigh raw material by above-mentioned parts by weight, be uniformly mixed to get.
Embodiment 3: the preparation of biological toner
Raw material composition:
60 parts of red phaffia rhodozyma spray powder, 12 parts of red phaffia rhodozyma freeze-dried powder, 8 parts of lactobacillus-fermented product, antibacterial peptide 8
Part, 2 parts of mountain flour, 1.5 parts of glutamine.
Preparation method is the same as embodiment 2.
Embodiment 4: the preparation of biological toner
Raw material composition:
80 parts of red phaffia rhodozyma spray powder, 8 parts of red phaffia rhodozyma freeze-dried powder, 12 parts of lactobacillus-fermented product, antibacterial peptide 4
Part, 4 parts of mountain flour, 0.5 part of glutamine.
Preparation method is the same as embodiment 2.
Embodiment 5: application of the biological toner in aquaculture of aquatic animal
1 materials and methods
The raising of 1.1 experimental animals
Test is temporarily supported in indoor aquarium (0.9m × 0.5m × 0.5m), is had with color crucian carp, average weight (9 ± 0.5) g
Effect water volume is 100L.Using the tap water of abundant aeration dechlorination as water source, water temperature is controlled in (24 ± 1) DEG C, is persistently filled for 24 hours
Oxygen；Day feeding volume is the 1.5% of fish body weight, and early, middle and late timing is fed.Test fish is pressed into weight after the adaptation training of 14d
4 groups, respectively control group (CK), test group I (Y I), test group II (Y II), competing product group (L) are randomly divided into, every group of 25 tails divide
It is not sampled in 14d, 28d, 42d, 56d, 60d.
The additive amount (two dosage groups that Y I, Y II are biological toner prepared by the embodiment of the present invention 2) of 1.2 toners
The biological toner additive amount (per kilogram basal diet) of table 7
Note: the competing product group of L is the sterling of chemical synthesis astaxanthin.
The adding method of 1.3 toners
It is uniformly sprayed by the additive amount of table 7, after toner is dissolved with water in color crucian carp complete feed, ventilation to half
It saves after dry to 4 DEG C of refrigerators.
The index of 1.4 measurements
Chromogenic indicator: the measurement of skin, scale, fin ray total carotinoid, the measurement of skin coloration, chromatophore meter
Number；
Growth indexes: weight, liver body index；
Immune performance: index and spleen index, middle renal index；
1.5 index determining methods
Liver body index: liver pancreas weight/weight；
Index and spleen index: spleen weight/weight；
Chromatophore counts: finding chromatophore under microscope 40x, counts chromatophore number under a visual field and counted
Number.
Coloration: using colorimetric card colorimetric, records chromatic value；
The measurement of astaxanthin: dimethyl sulfoxide method；
2 test results
2.1 chromogenic indicator
2.1.1 total carotinoid
Scale, skin, fin ray total carotinoid assay are as a result, be shown in Table 8.
8 scale of table, skin, fin ray total carotinoid content
Note: same column has same alphabetical expression difference not significant (P > 0.05), has different alphabetical expression significant difference (P <
0.05)。
As can be seen from Table 8, scale is first to reduce then to rise by a small margin again to the accumulation law of total carotinoid,
In 14d, test group I, test group II, competing product group and control group are all remarkably higher than control group in 60d without significant difference；
Skin appears in 28d to the significant sexual intercourse of total carotinoid, until only adding biological toner group when 60d is significantly higher than control
Group；The significant sexual intercourse of fin ray total carotinoid also appears in 28d, until only to add high dose biology toner group significant by 60d
Higher than control group.Analysis reason may be to launch into albefaction bucket early period, similar since fish have automatic regulation of the body color mechanism
In the protective coloration of animal, the color of automatic closeness to life environment causes pigment accumulation ability to decline, albefaction trend occurs, due to
The long-term addition situation of pigment takes a turn for the better, but since in 42d, the time enters mid-November, temperature is begun to decline, and is ingested
Reduce, biological toner of intake etc. reduces, so pigment accumulation have it is on a declining curve.It is overall, from the comparison between each group
From the point of view of, biological toner addition group effect is better than competing product group, and the astaxanthin of effect stability, competing product group addition belongs to chemical synthesis
It is single to be chemical synthesis astaxanthin ingredient though itself has whitening power for astaxanthin, and had in synthesis process it is some can not
The harmful substance of removal, long-time service will affect fish body function, cause pigment accumulation effect poor.
2.1.2 skin coloration and chromatophore count
9 skin coloration of table and chromatophore count mean value
As can be seen from Table 9, it is fed through 60d, test group I (Y I), the skin coloration mean value of test group II (Y II), color
Plain cell mean value is above control group；From Fig. 1, Fig. 2 it is also seen that chromatophore individual and chromatophore density are above pair
According to group, sector area is formed by also greater than control group, illustrates that this product can promote chromatophore development, proliferation.
2.2 growth indexes
2.2.1 weight
10 each group changes of weight of table
As can be seen from Table 10, test group I, test group II, competing product group and control group there are no significant difference, but test
Group I and II weight of test group are continuous upward trends, and only an inflection point occur in competing product group and control group, illustrate addition biology
Toner group either can preferably improve weight and effect stability, high dose biology toner in low dosage or high dose group
Group effect is better than low dose group.
2.2.2 liver body index
The variation of 11 each group liver body index of table
As can be seen from Table 11, that there are no significant is poor for test group I, test group II, the liver body index of competing product group and control group
It is different, but it is in continuous upward trend, and biological toner high dose group effect is optimal.
2.3 immune indexes
12 each group immune indexes of table
As can be seen from Table 12, test group I, test group II, competing product group and control group there are no significant difference, addition biology
Toner group effect is slightly better than competing product group and control group, and the possible cause that inflection point occurs in competing product group is: artificial synthesized astaxanthin contains
There is by-product, the long-time service of by-product may result in fish body immune organ and be damaged, and limit Development of Immune Organs.And biology increases
The natural astaxanthin and nutritional ingredient that toner itself contains can promote Development of Immune Organs, play the work for improving immunity of organisms
With.
2.4 digestion indexs
2.4.1 enteron aisle digestive ferment
13 each group enteron aisle of table digests enzymic change
As can be seen from Table 13, biological toner group is added in 60d, and proteinase activity is significantly higher than control group and competing product
Group, amylase only add high dose biology toner group and are significantly higher than other each groups, and illustrating to add biological toner can be significant
It improves digestive enzyme activity and increases significant effect with dosage.
Claims (7)
1. a kind of biology toner, which is characterized in that its raw material composition are as follows: 60-80 parts of red phaffia rhodozyma spray powder, red Fife's ferment
Female freeze-dried powder 8-12 parts, 8-12 parts of lactobacillus-fermented product, 4-8 parts of antibacterial peptide, 2-4 parts of mountain flour, 0.5-1.5 parts of glutamine；
Wherein, the red phaffia rhodozyma spray powder and red phaffia rhodozyma freeze-dried powder, be by red phaffia rhodozyma in yeast culture medium
It carries out fermented and cultured and obtains fermentation liquid, fermentation liquid is prepared through spray drying and freeze-drying respectively；
The red phaffia rhodozyma is the yeast strain of high-yield astaxanthin function, strain number CICC33064；
The lactobacillus-fermented product is that Pediococcus pentosaceus PP (PediococcuspentosaceusPP) fermented culture obtains
Tunning；The deposit number of Pediococcus pentosaceus PP is CCTCCNO:M2012029.
2. biology toner as described in claim 1, which is characterized in that its raw material composition are as follows: red phaffia rhodozyma spray powder 70
Part, 10 parts of red phaffia rhodozyma freeze-dried powder, 10 parts of lactobacillus-fermented product, 6 parts of antibacterial peptide, 3 parts of mountain flour, 1 part of glutamine.
3. biology toner as described in claim 1, which is characterized in that the raw material of the yeast culture medium forms are as follows: grape
Sugared 1.5%, peptone 1.0%, yeast extract 0.5%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.02%, sodium chloride 2.0%, chlorine
Change calcium 0.02%, glutamic acid 0.06%；It is mass percent.
4. the preparation method of the described in any item biological toners of claim 1-3, which is characterized in that by red phaffia rhodozyma spray drying
Powder, red phaffia rhodozyma freeze-dried powder, lactobacillus-fermented product, antibacterial peptide, mountain flour and glutamine be uniformly mixed to get.
5. the described in any item biological toners of claim 1-3 are applied in aquaculture of aquatic animal.
6. application as claimed in claim 5, which is characterized in that application method are as follows: by the described in any item lifes of claim 1-3
Object toner is added in aquatic animal feed in the ratio that mass fraction is 1-3 ‰, is one within 30 days and is fed the period or long-term
It feeds.
It is by adding in aquatic livestock basal feed by mass fraction for 1-3 ‰ 7. a kind of aquaculture of aquatic animal is hyperchromic to use feed
The described in any item biological toners of claim 1-3 are added to be prepared.
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| CN108094754A (en) * | 2017-11-24 | 2018-06-01 | 镇江华优生态农业科技有限公司 | A kind of young extremely detains the mixed feed in crab stage and preparation method thereof |
| CN112515053A (en) * | 2020-11-02 | 2021-03-19 | 无锡泓宝微生物科技有限公司 | A QINGSU bacterium for enhancing pigment and immunity of aquatic animal |
| CN112617027A (en) * | 2020-12-31 | 2021-04-09 | 上海泓宝绿色水产股份有限公司 | Bdelloecium bdellovii and preparation method and use method thereof |
| CN114431333B (en) * | 2021-12-24 | 2024-04-02 | 青岛尚德生物技术有限公司 | Phaffia rhodozyma liquid compound preparation and preparation method and application thereof |
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| CN103865798A (en) * | 2014-03-18 | 2014-06-18 | 厦门汇盛生物有限公司 | Method for wall breaking of phaffia rhodozyma through enzymolysis and application thereof |
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