CN106010990B - A kind of rhodotorula mucilaginosa and its fermentation culture medium and application - Google Patents
A kind of rhodotorula mucilaginosa and its fermentation culture medium and application Download PDFInfo
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- CN106010990B CN106010990B CN201610317014.5A CN201610317014A CN106010990B CN 106010990 B CN106010990 B CN 106010990B CN 201610317014 A CN201610317014 A CN 201610317014A CN 106010990 B CN106010990 B CN 106010990B
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K36/07—Basidiomycota, e.g. Cryptococcus
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
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- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
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- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Abstract
Disclosed herein is a kind of rhodotorula mucilaginosa and its fermentation culture medium and application, the rhodotorula mucilaginosa are as follows: rhodotorula mucilaginosa TZR2014, CCTCC NO:M2015574.Liquid state fermentation is carried out to it using fermentation medium provided by the invention, the biomass of fermentation liquid is 18g/L, and the yield of carotenoid is 36 mg/L.Rhodotorula mucilaginosa of the invention has an anti-Staphylococcus aureus, the effect of stomach juice-resistant and cholate, also contains pyruvic acid in solid-state fermentation content, and pyruvic acid has and accelerates fat consumption, improves muscle yield, phagocytosis interior free yl and the systematic function for inhibiting free radical.Therefore rhodotorula mucilaginosa provided by the invention can be used for the feed addictive of beasts, birds and aquatic products animal, have wide application prospect.
Description
Technical field
The present invention relates to field of biotechnology, it is more particularly to a kind of rhodotorula mucilaginosa and its fermentation culture medium and application.
Technical background
Rhodotorula mucilaginosa category basidiomycetes, bacterium colony are cream-colored, pink to red mucus shape, are simple pseudohyphas, carefully
Born of the same parents' shape is circle, is nutrition body cell gemmation.Rich in astaxanthin, carotenoid and beta glucan.Carotenoid
It is a kind of crocus fat-soluble compound, the double effects with nutritional supplement and food color.It cannot be closed in animal body
At carotenoid, it is necessary to be provided by food, it is the important sources of vitamin A in animal body, there is nourishing liver and improving eyesight, can
Treat yctalopia.Carotenoid is also a kind of antioxidant, and then prevents a variety of degenerative disorders caused by aging and aging.
Meanwhile there is significant effect in anticancer, prevention cardiovascular and cerebrovascular disease, cataract.Carotenoid is as a kind of natural pigment
In animal husbandry more and more attention has been paid to.Carotenoid is made greatly improve the appearance and yolk of carcass in poultry deposition
Colorimetric fans series, improves yolk color.Pyruvic acid as dietary supplements, have accelerate fat consumption, lose weight, improve it is thin
Meat rate and other effects；There is special protective effect to heart, the efficiency of cardiac muscle can be enhanced, reduces heart disease or heart locally lacks
It is damaged caused by blood；Meanwhile pyruvic acid has phagocytosis interior free yl and inhibits the remarkable results such as the generation of free radical.Utilize glue
Rhodotorula fermenting and producing astaxanthin, carotenoid and beta glucan have nutritive value simple, and cultivation cycle is short, at low cost
It is honest and clean, the advantages of thallus is nontoxic and protein rich in, there is important research significance and development prospect.Southwestern China area
With heavy rainfall, air humidity is big, and the growth of benign climate environmental microorganism, barms are resourceful, but rhodotorula strain provides
Excavate limited in source.Based on this, in order to enrich rhodotorula mucilaginosa strain resources bank, the present invention is directed to excavate the southwest with economic value
Regional rhodotorula mucilaginosa strain resource, studies its liquid and solid state fermentation cultural method, and tunning is supported applied to beasts, birds and aquatic products
It grows in industry.
Summary of the invention
The purpose of the present invention is to provide a kind of rhodotorula mucilaginosa TZR2014, the yeast be rich in amino acid, carotenoids can be produced
Element and pyruvic acid, have anti-Staphylococcus aureus, and the effect of stomach juice-resistant and cholate can be applied to beasts, birds and aquatic products cultivation.The bacterium
Strain was sent on September 24th, 2015 to China typical culture collection center preservation, classification naming: rhodotorula mucilaginosa
(Rhodotorula muciladinosa)TZR2014, address: Wuhan, China Wuhan University, deposit number: CCTCC NO:M
2015574。
It is another object of the present invention to provide a kind of rhodotorula mucilaginosa TZR2014Fermentation liquid culture and solid-state training
Object is supported, fermentating formula of the invention is simple, low in cost.Utilize fermentation liquid fermentation rhodotorula mucilaginosa TZR2014, can mass production
Barms, culture carotenoid containing high yield and pyruvic acid.
Final object of the present invention is the provision of a kind of rhodotorula mucilaginosa TZR2014Application, including the red ferment of the glue
Application of female fermentation liquid culture or solid state rheology object in animal husbandry and fishery aquaculture.
In order to achieve the above object, the present invention takes following technical measures:
Rhodotorula mucilaginosa is separating obtained from the soil in Chongqing City citrus research institute shaddock garden；The bacterial strain is in September 24 in 2015
Day is sent to China typical culture collection center preservation, classification naming: rhodotorula mucilaginosa (Rhodotorula muciladinosa)
TZR2014, deposit number: CCTCC NO:M 2015574；Address: Wuhan, China Wuhan University.
The morphological feature of bacterium colony: bacterium colony is rounded or oval, thick, and color is pink, and bacterium colony is big and thick, is easy to
It provokes, glimmering luster.
Rhodotorula mucilaginosa physiological and biochemical property
| Galactolipin | Sucrose | Fine disaccharides | Raffinose | Sorbierite | Melezitose | Inositol | Trehalose |
| + | + | - | + | + | + | - | + |
| Acetamide | Mannitol | Rhamnose | Melibiose | Xylose | Ribose | Glycerol | I is sugared |
| - | + | - | - | + | + | + | + |
+: it is positive；: it is negative
A kind of rhodotorula mucilaginosa TZR2014Liquid state fermentation culture medium, comprising: glucose 3-5%, peptone 1-2%, yeast powder
0.5-1.5%, pH6.0, remaining is water, is above mass ratio.
In scheme described above, it is preferred that the formula of liquid state fermentation culture solution includes: glucose 5%, peptone 1%,
Yeast powder 1.5%, remaining is water, pH 6.0；The biomass of rhodotorula mucilaginosa, pyruvic acid and carotenoid output reach most at this time
Greatly.
A kind of rhodotorula mucilaginosa TZR2014Solid-state fermentation culture medium, comprising: wheat bran 15-30%, dregs of beans 6-8%, corn flour 1-
3%, corn pulp 3-5%, rice bran 2-4%, ammonium sulfate 0.5-1%, dipotassium hydrogen phosphate 0.03-0.08%, magnesium sulfate 0.08-
0.12%, remaining is tap water, is above mass ratio, and the solid-state fermentation culture medium is through 121 DEG C of high-temperature steam sterilizations
20min, being cooled to 40 DEG C can access rhodotorula mucilaginosa TZR2014。
In scheme described above, the formula of preferred solid-state fermentation culture medium includes: wheat bran 22.39%, dregs of beans
7.31%, corn flour 2.32%, corn pulp 4.00%, rice bran 3.12%, ammonium sulfate 0.69%, dipotassium hydrogen phosphate 0.053%, sulphur
Sour magnesium 0.103%, remaining is tap water, and pH6.0 is mass ratio above；In this formula, the biomass of rhodotorula mucilaginosa, third
Ketone acid and carotenoid output reach maximum.
A kind of rhodotorula mucilaginosa TZR2014Application, fermentation liquid culture or solid state rheology object including the rhodotorula mucilaginosa
Application in animal husbandry and fishery aquaculture can be used for preparing the bacterium for inhibiting staphylococcus aureus including the rhodotorula mucilaginosa
Agent, for fowl poultry kind or the feed addictive of aquatic product animal.
Compared with prior art, the invention has the following advantages that
1, the rhodotorula mucilaginosa announced is the gained from ocean point, and rhodotorula mucilaginosa of the invention is from citrus research institute citrus
It is separating obtained in Orchard Soil, such rhodotorula mucilaginosa is confirmed without any toxicity and side effects by animal safety test；
2, after rhodotorula mucilaginosa of the invention is fermented, carotenoid content reaches 36mg/L in fermentation liquid.
3, rhodotorula mucilaginosa of the invention has anti-Staphylococcus aureus, the effect of stomach juice-resistant and cholate.
4, rhodotorula mucilaginosa obtained by solid state fermentation of the present invention can also produce pyruvic acid.And pyruvic acid has acceleration fat consumption, mentions
High muscle yield, phagocytosis interior free yl and the systematic function for inhibiting free radical.
5, it is aquatic feed for domestic animal raw material that solid state fermentation culture is raw materials used, low in cost.It is rich in culture
Essential amino acid, carotenoid, pyruvic acid, growth of animal and product quality can be improved by generating applied to animal.
Detailed description of the invention
Fig. 1 is rhodotorula mucilaginosa bacterium colony aspect graph in embodiment 1.
Fig. 2 is rhodotorula mucilaginosa cellular morphology figure in embodiment 1.
Specific embodiment
Technical solution of the present invention is if not otherwise specified the ordinary skill in the art；The reagent or material are such as
It is not specified, derive from commercial channel.
Embodiment 1: strain separating and identification
Strain isolation
Soil 100g is adopted from citrus research institute, Chongqing City citrus orchard, takes the soil sample of 10g in equipped with 100ml sterile water
In 250ml triangular flask, a moment is stood after vibrating in vortex instrument, the physiological saline of the bacteria suspension and 9ml that take 1ml mixes, then takes
The bacteria suspension of 1ml and the physiological saline of 9ml mix, and so on, different concentration gradients, gradient 10 are set-1、10-2、10-3、10-4、10-5, successively take the bacteria suspension of 200 μ l to be coated with and be inoculated on separation plate, 28 DEG C of 2~3d of culture are placed in, according to bacterium colony
(bacterium colony is rounded or oval, thick, and color is pink, and bacterium colony is big and thick, is easy to choose for form picking typical case single colonie
Rise, glimmering luster), Rhodotorula mucilaginose is gone out by microscopy scalping.
PDA culture medium: potato 20g, glucose 20g, agar 20g, water 1000ml, natural pH, sterilize by 115 DEG C
20min。
Secondary screening and identification
It is screened by Oxford cup bacteriostatic test to staphylococcus aureus (G+) the good glue of pathogenic bacteria growth inhibition performance is red
Yeast is identified by Physiology and biochemistry and 26S rDNA D1/D2, is finally screened to one plant of production beta carotene Rhodotorula mucilaginose, life
Entitled RhodotorulamuciladinosaTZR2014.The bacterium has been sent to Chinese Typical Representative culture guarantor on September 24th, 2015
Hiding center carries out preservation, classification naming: rhodotorula mucilaginosa (Rhodotorulamuciladinosa) TZR2014, address: Wuhan, China
Wuhan University.
TZR2014Bacterial strain specifically has the physiological and biochemical property of typical rhodotorula mucilaginosa: the bacterial strain breeds life in PDA culture medium
The long bacterium colony formed is subcircular, thick, and color is pink, and bacterium colony is big and thick, is easy to provoke, glimmering luster (Fig. 1, Fig. 2).
Physiological and biochemical property are as follows: gelatin liquefaction.It is ferment galactose, sucrose, glucose, raffinose, sorbierite, melezitose, trehalose, sweet
Reveal alcohol, xylose, ribose, glycerol and arabinose；Fructose and maltose, azymic fibre disaccharides, inositol, acetamide, rhamnose and
Melibiose.28~32 DEG C of optimum temperature, 50 DEG C of maximum temperature.
So far, applicant or one plant of rhodotorula mucilaginosa was obtained, which sent on September 24th, 2015 to Chinese Typical Representative culture
Object collection preservation, classification naming: rhodotorula mucilaginosa (Rhodotorulamuciladinosa) TZR2014, deposit number:
CCTCC NO:M 2015574；Address: China, Wuhan, Wuhan University.
Embodiment 2:
Rhodotorula mucilaginosa (Rhodotorulamuciladinosa) TZR2014Large-scale fermentation:
By the rhodotorula mucilaginosa TZR of preservation2014It is inoculated into PDA culture medium, is placed in 28 DEG C of constant incubator culture 36h,
One oese rhodotorula mucilaginosa of picking is in seed culture fluid (mass fraction, %): glucose 2%, peptone 1%, yeast powder 1%,
Natural pH, remaining is distilled water) in, it is cultivated for 24 hours on 28 DEG C, the isothermal vibration device of 200rpm.
500 liters of fermentation tank capacities are 65% (V/V), and liquid state fermentation culture medium includes (mass fraction, %): glucose
5%, peptone 1%, yeast powder 1.5%, pH6.0 are cooled to 40 DEG C, connect thereto through 121 DEG C of high-temperature steam sterilization 20min
The seed liquor of 10% cell age of kind for 24 hours, at 28 DEG C, 200r/min ferments after 96h, and the biomass of fermentation liquid is 18g/L, carotenoids
The yield of element is 36mg/L.
After putting tank, filter plate is concentrated into original volume 1/8 through filter press, in 0.2~0.3MPa of gauge pressure, and spray drying obtains
(effective bacterial content of every gram of dry powder is 3.2 × 10 to active dry yeasr dry powder11Cfu/g), it is used for embodiment 10 and 11.
Embodiment 3:
Rhodotorula mucilaginosa TZR2014The content of protein and amino acid
Bacterium mud is collected by centrifugation in fermentation liquid in embodiment 2, drying obtains dry bacterium powder at 55 DEG C.
Determined amino acid: dry bacterium powder 120mg is accurately weighed in test tube, 10ml is added in the test tube of Xiang Shengyou sample
16mol hydrochloric acid, concussion mix.With alcohol blast burner drawing and attenuating to 4-6mm at 1/3 under the test tube, tube sealing after ten minutes is vacuumized.Place
The test tube managed is set in 110 DEG C of constant temperature ovens sand-bath and is hydrolyzed 22 hours, takes out and is cooled to room temperature, shakes up filtering, take 1ml filtrate in
In 50ml beaker, it is evaporated filtrate with 60 DEG C of waters bath with thermostatic control, 4 times of 0.02mol hydrochloric acid is added, uses day with 0.22 μm of membrane filtration
The vertical full-automatic amino-acid analyzer measurement of L-8800 type.Crude protein content is measured using Kjeldahl's method (GB5009.5-2003).
Rhodotorula mucilaginosa dry bacterium powder contains 35.75% crude protein, 2.03% aspartic acid, 1.14% threonine, 1.19% ammonia
Acid, 2.29% glutamic acid, 1.33% glycine, 2.02% alanine, 0.19% cysteine, 1.25% valine, 0.39%
Methionine, 1.05% isoleucine, 1.77% leucine, 0.652 tyrosine, 1.049% phenylalanine, 1.259% lysine,
0.582% histidine, 1.74% arginine %, 1.16% proline.
Embodiment 4:
Rhodotorula mucilaginosa TZR2014Solid state fermentation:
Rhodotorula mucilaginosa is inoculated into PDA culture medium, 28 DEG C of constant incubator culture 36h, one oese of picking are placed in
Rhodotorula mucilaginosa is in seed culture fluid (each composition quality score (%): glucose 2%, peptone 1%, yeast powder 1%, natural pH)
In, it is cultivated for 24 hours on 28 DEG C, the isothermal vibration device of 200rpm.
Solid-state fermentation culture medium includes (mass fraction, %): wheat bran 22.39%, dregs of beans 7.31%, corn flour 2.32%,
Corn pulp 4.00%, rice bran 3.12%, ammonium sulfate 0.69%, dipotassium hydrogen phosphate 0.053%, magnesium sulfate 0.103%, remaining is certainly
Water, pH6.0 are cooled to 40 DEG C through 121 DEG C of high-temperature steam sterilization 20min.The seed liquor 7.5% of inoculation cell age for 24 hours thereto
(mass ratio), ferment 72h in 28 DEG C of constant temperature biochemical cultivation case, and it is dry preceding containing 10 finally to measure every g solid state fermentation culture9
A rhodotorula mucilaginosa, 190.2 μ g carotenoid, 1.32mg pyruvic acid.Solid-state fermentation content is made rhodotorula mucilaginosa through 40 DEG C of dryings and raises
Feed additives are used for embodiment 12.
Embodiment 5:
Rhodotorula mucilaginosa TZR2014Resistance detection
Hot tolerance: take rhodotorula mucilaginosa growth latter stage medium centrifugal, 0.9% brine twice after again
Suspend (1~2 × l0 of viable bacteria amount9Cfu/ml after) handling 15min in 35,45,55,65,75,85,95 DEG C of water-baths, rapid flowing water
It is cooling, survival rate is then measured using dilution-plate method.The result shows that the strain highest tolerable temperature is 55 DEG C (being shown in Table 1).
Survival rate of 1 rhodotorula mucilaginosa of table under treatment of different temperature
Gastric fluid-resistant ability: taking mass fraction is that 10% hydrochloric acid 16.4ml adds distilled water to dilute, and makes pH value 2.0, often
1g pepsin is added to obtain artificial stomach after completely dissolution with 0.22 μm of filtering with microporous membrane degerming in 100ml hydrochloric acid solution
Liquid.Rhodotorula mucilaginosa growth final stage culture centrifugation is taken, 0.9% brine suspends again after twice, with 107Cfu/ml is whole
Concentration is inoculated in simulated gastric fluid, and concussion mixes, and 37 DEG C of water-baths are incubated for, and is sampled every 0.5h and is carried out count plate.The result shows that
Rhodotorula mucilaginosa is resistant to gastric juice up to 2 hours (being shown in Table 2).
2 rhodotorula mucilaginosa of table handles the variation of viable bacteria amount under the conditions of simulation hydrochloric acid
| It handles time (h) | 0 | 0.5 | 1 | 1.5 | 2 |
| Bacterium amount (cfu/ml) | 1.06×l09 | 6.90×l08 | 6.31×l08 | 5.83×l08 | 5.10×l08 |
Pig cholate tolerance: preparing additional addition 1.0%, 2.0%, 3.0%, 4.0%, 5.0% (w/v) Pig cholate
Seed culture fluid.1ml is taken to take rhodotorula mucilaginosa bacterium solution (1~2 × l08Cfu/mL), it is inoculated in 9mL various concentration high bile salt concentiration kind
In sub- culture solution, after 30 DEG C of culture 12h, measurement culture solution turbidity variation determines thalli growth situation.The result shows that rhodotorula mucilaginosa
Being resistant to Pig cholate mass fraction is 3% (being shown in Table 3).
Table 3: the influence that various concentration Pig cholate grows rhodotorula mucilaginosa
Note: ++ well-grown is represented ,+represent growth generally ,-represent and cannot grow.
Embodiment 6:
The red glue mother strains bacteriostasis detection of glue
Sterilized NB agar medium is heated to melt completely, is poured in culture dish, every ware 15ml (lower layer), to it
Solidification.The training solidified is added in addition, the NB culture medium of thawing is cooled to 50 DEG C or so and is mixed into pharmacosensitive test bacteria culture fluid 5ml
It supports on base (upper layer) to be solidified.With sterile working media surface directly vertically put Oxford cup (internal diameter 6mm, outer diameter 8mm,
High 10mm) round tubule, the both ends of pipe want smooth), gently pressurize, make itself and culture medium contact tight, be added in cup
Measuring samples (fermentation liquid), Oxford cup can generally fill 200 microlitres, not make its excessive.It is small to fill it up with 37 DEG C of culture 16-18 of postposition
When, it observes as a result, inhibition zone is directly measured with ruler.The result shows that rhodotorula mucilaginosa have the function of inhibit staphylococcus aureus (see
Table 4).
Antibacterial circle diameter of 4 rhodotorula mucilaginosa of table to different pathogenic bacteria
| Pathogenic bacteria | Escherichia coli | S. pullonum | Staphylococcus aureus |
| Inhibition zone (mm) | 0 | 0 | 20 |
Embodiment 7:
Mouse safety test
The kunming mice of 60 weight 20g or so is randomly divided into 3 groups, every group 20, half male and half female.Wherein do sky for one group
White control, experimental group one every is pressed 1 × 109Cfu/kg mouse weight (100 times of routine dose) gavages rhodotorula mucilaginosa in continuous one week
Fermentation liquid, two disposable celiac of experimental group inject the bacteria suspension (1 × 10 of 0.2ml9cfu/ml).Observation one week, record each group are small
The growth of mouse and death condition (table 5).All mouse feeding experiments are in drying, cleaning, draughty indoor completion.As a result
Show: after a week, 3 groups, without death, are had an injection and organize mouse food-intake reduction of having an injection for first day, apathetic, are restored just within the 3rd day
Often, the results are shown in Table 5.
Table 5: mouse experiment weighing results
| Experimental group | Initial weight (g) | Weighing (g) in 3 days | Weighing (g) in 7 days |
| Control group | 19.26±0.7 | 20.91±1.67 | 23.78±1.94 |
| Experimental group one | 19.24±0.81 | 22.36±0.19 | 26.93±1.91 |
| Experimental group two | 19.28±0.6 | 20.31±2.62 | 22.00±1.93 |
Experimental group one is stomach-filling group, and experimental group two is group of having an injection.
Embodiment 8:
The early-weaned piglets of microbial inoculum are tested
It chooses similar in weight, with 21 age in days piglet of batch 40, is divided into 4 groups, every group 10, one group is control group, real
Group A every is tested by percentage of liveweight 108The bacterium amount of cfu/kg is mixed water and is gavaged, and experimental group B every is pressed percentage of liveweight 109The bacterium amount of cfu/kg is mixed
Water gavages, and experimental group C every is pressed percentage of liveweight 1010The bacterium amount of cfu/kg mixes water and gavages experimental group to be filled once daily, continuous to fill 7 days,
Observation 21 days.It is freely eaten, every day entry spirit and feeding situation are weighed at the end of experiment.Germicidal efficacy by 21 days exists
Large dosage continuously gavages lower without abnormal response, and feeding and growing state are normal, and rhodotorula mucilaginosa group has obvious growth promotion
Effect (table 6).
Table 6: piglet gavages various dose TZR2014Influence to weight gain
| Group | First weight kg | Opisthosoma weight kg | Increase weight kg |
| Blank group | 5.90 | 9.93 | 4.03 |
| Experimental group A | 6.08 | 10.91 | 4.83 |
| Experimental group B | 6.03 | 11.87 | 5.84 |
| Experimental group C | 5.91 | 10.62 | 4.71 |
Embodiment 9:
The pregnant sow of microbial inoculum is tested
Same batch pregnant sow 20 is chosen, is divided into 2 groups, every group 10, one group is control group, and another group is experiment
Group；Observation period is every and produces the previous moon to sucking pig wean；Control group diet daily ration；Experimental group by pig weight 2 ×
109The bacterium amount of cfu/kg is added to feeding in feed diet, daily to observe sow spirit and excrement situation, at the beginning of postpartum claims sucking pig
Weight.Observe that experimental group prevention of sow constipation and the condition of production are obviously improved, control group be averaged sucking pig initial weight be 1.31 kilograms, litter size
Be 8.9, and experimental group be averaged sucking pig initial weight be 1.49 kilograms, litter size 10.3.
Embodiment 10:
Application effect of the bacterium powder in laying hen and laying duck cultivation.
Select body condition close, brown laying hen 200, sea orchid 39 week old, test period is 90 days, and first 5 days are preliminary trial period, after
It is positive the examination phase within 85 days.It is classified as 2 processing groups, 5 repetitions of each processing group, each repetition 20 at random.I.e. 1 control
Group, 1 test group.Control group fed basal diet, test group are to be added to the basic day of 1.5% active rhodotorula mucilaginosa dry powder
Grain.
Selection body condition is close, gold of 40 week old determine laying duck 200, and test period is 90 days, is within first 5 days preliminary trial period, and rear 85
It is positive the examination phase.It is classified as 2 processing groups, 5 repetitions of each processing group, each repetition 20 at random.That is 1 control group, 1
A test group.Control group fed basal diet, test group are the basal diet for being added to 2% active rhodotorula mucilaginosa dry powder.
The results are shown in Table 7.
7 rhodotorula mucilaginosa TZR of table2014Growth-promoting effect in laying hen and laying duck cultivation
Embodiment 11:
Application effect of the bacterium powder in Roche natural pond juvenile prawn cultivates.
Test select the hatching of same batch, Individual Size uniformly, physical health, long body is about 4.2cm, weight 1.8g or so
200 tail of Roche natural pond juvenile prawn, be randomly divided into 2 processing groups, 5 repetitions of each processing group, each 20 tail shrimp of repetition, i.e., 1 is right
According to group, 1 test group.Control group fed basal diet, test group are the active rhodotorula mucilaginosa dry powder that feeding is added to 4.0%
Basal diet.Weighing record initial weight, is raised in 200L automatic inflating glass fibre bucket.Experimental period 55d.Periodically increase during test
Oxygen；Dissolved oxygen 4.5-5.0g/L, in due course replacement water source.Water temperature range is at 22~28 DEG C；Water pH value is 7.6 or so.Exist daily
08:00,17:00,22:00 bait throwing in record daily ration, feeding quantity, and according to ingest situation and the water temperature variation adjustment in due course of shrimp, day bait throwing in
Amount is the 6%~8% of weight, feeds close to being satiated with food, does not stay residual bait as far as possible, the results are shown in Table 8.
8 rhodotorula mucilaginosa TZR of table2014Growth-promoting effect in the cultivation of Roche natural pond juvenile prawn
| Experiment process | Specific growth rate (%/d) | Food conversion | Survival rate |
| Control group | 3.63 | 1.34 | 61% |
| Test group | 3.82 | 1.21 | 69% |
Embodiment 12:
Application effect of the solid-state fermentation content in laying hen and laying duck cultivation.
Select body condition it is close, Roman egghen 200 39 week old, test period be 90 days, first 5 days be preliminary trial period, rear 85
It is positive the examination phase.It is classified as 2 processing groups, 5 repetitions of each processing group, each repetition 20 at random.That is 1 control group, 1
A test group.Control group fed basal diet, test group are to be added to the basic day of 5.0% active rhodotorula mucilaginosa solid-state fermentation content
Grain.
Selection body condition is close, gold of 40 week old determine laying duck 200, and test period is 90 days, is within first 5 days preliminary trial period, and rear 85
It is positive the examination phase.It is classified as 2 processing groups, 5 repetitions of each processing group, each repetition 20 at random.That is 1 control group, 2
A test group.Control group fed basal diet, test group are to be added to the basic day of 6.0% active rhodotorula mucilaginosa solid-state fermentation content
Grain.
The results are shown in Table 9.
9 TZR of table2014Growth-promoting effect in laying hen and laying duck cultivation
Root Ju embodiment 10 and 12 is as a result, TZR2014Liquid state fermentation object and solid-state fermentation content can improve laying hen and egg laying duck
Growth performance, effect is suitable, but for feed cost solid-state fermentation content fermentation it is raw materials used cheap compared with liquid state fermentation object, because
This solid-state fermentation content feeding cost is lower.
Claims (8)
1. a kind of rhodotorula mucilaginosa, it is characterised in that: the rhodotorula mucilaginosa are as follows: rhodotorula mucilaginosa (Rhodotorula
muciladinosa)TZR2014；Deposit number are as follows: CCTCC NO:M 2015574.
2. rhodotorula mucilaginosa described in claim 1 is preparing the application in carotenoid.
3. rhodotorula mucilaginosa described in claim 1 inhibits the application in staphylococcus aureus preparation in preparation.
4. rhodotorula mucilaginosa described in claim 1 is preparing the application in livestock and poultry animal feed addictive.
5. rhodotorula mucilaginosa described in claim 1 is preparing the application in aquatic animal feed additive.
6. application according to claim 4, it is characterised in that: the rhodotorula mucilaginosa improves laying hen or laying duck at the same time
The application of laying rate and egg material than in.
7. application according to claim 4, it is characterised in that: the rhodotorula mucilaginosa is in promoting fowl poultry kind growth of animal
Application.
8. application according to claim 5, it is characterised in that: the rhodotorula mucilaginosa is in promoting aquatic product growth of animal
Application.
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| CN107653195A (en) * | 2017-09-14 | 2018-02-02 | 贵州东智生物科技有限公司 | A kind of method that rhodotorula mucilaginosa is separated from fermenting black garlic |
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| RU2712729C1 (en) * | 2018-10-29 | 2020-01-30 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Горский государственный аграрный университет" | Yeast strain rhodotorula mucilaginosa ayr 8-2018 vkpm-y-4342 - producer of microbial protein and carotenoids |
| CN109548984A (en) * | 2018-12-19 | 2019-04-02 | 江南大学 | A method of oil crops are processed using rhodotorula |
| CN109965136A (en) * | 2019-04-08 | 2019-07-05 | 南京晓庄学院 | A kind of rejected layer meat improved formulations, preparation method and its application |
| CN114921352B (en) * | 2022-05-10 | 2023-07-21 | 吉林省农业科学院 | Rhodotorula mucilaginosa JAASRY1, fermentation culture and application thereof |
| CN114794303B (en) * | 2022-05-10 | 2023-07-14 | 吉林省农业科学院 | Selenium-enriched rhodotorula mucilaginosa, selenium-enriched rhodotorula mucilaginosa feed additive, and preparation method and application thereof |
| CN114874940B (en) * | 2022-05-10 | 2023-07-14 | 吉林省农业科学院 | Composite microbial agent, fermented product thereof and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007120423A2 (en) * | 2006-03-20 | 2007-10-25 | Microbia Precision Engineering | Production of quinone derived compounds in oleaginous yeast and fungi |
| CN102894237A (en) * | 2012-11-08 | 2013-01-30 | 黑龙江省轻工科学研究院 | Method for preparing feed additive for red-feather serinus canaria |
| CN103409328A (en) * | 2013-06-09 | 2013-11-27 | 江南大学 | Rhodotorula mucilaginosa and application thereof in degradation and decoloring of dyes and production of carotenoid |
-
2016
- 2016-05-13 CN CN201610317014.5A patent/CN106010990B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007120423A2 (en) * | 2006-03-20 | 2007-10-25 | Microbia Precision Engineering | Production of quinone derived compounds in oleaginous yeast and fungi |
| CN102894237A (en) * | 2012-11-08 | 2013-01-30 | 黑龙江省轻工科学研究院 | Method for preparing feed additive for red-feather serinus canaria |
| CN103409328A (en) * | 2013-06-09 | 2013-11-27 | 江南大学 | Rhodotorula mucilaginosa and application thereof in degradation and decoloring of dyes and production of carotenoid |
Non-Patent Citations (1)
| Title |
|---|
| "胶红酵母（Rhodotorula muciladinosa）及其降解氨基甲酸乙酯的研究";起雅敏等;《工业微生物》;20131031;第43卷(第5期);第44-48页 * |
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