CN106190912A - Photosynthetic bacteria microbial inoculum and preparation method and application - Google Patents
Photosynthetic bacteria microbial inoculum and preparation method and application Download PDFInfo
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- CN106190912A CN106190912A CN201610584746.0A CN201610584746A CN106190912A CN 106190912 A CN106190912 A CN 106190912A CN 201610584746 A CN201610584746 A CN 201610584746A CN 106190912 A CN106190912 A CN 106190912A
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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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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
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Abstract
The invention discloses a kind of photosynthetic bacteria microbial inoculum and preparation method and application.The photosynthetic bacteria microbial inoculum of the present invention, active component be Rhodopseudomonas palustris (Rhodopseudomonas palustris), the bacterial strain number of described Rhodopseudomonas palustris (Rhodopseudomonas palustris) is 981, and it is at the numbered CGMCC No.10802 that registers on the books at China Committee for Culture Collection of Microorganisms's common micro-organisms center.Active component Rhodopseudomonas palustris 981 reproduction speed of this photosynthetic bacteria microbial inoculum is fast, in bean sprouts medium (fermentation medium B) 30 DEG C, cultivate under the illumination condition that intensity of illumination is 1500Lx of 24 hours every days 3 days thalline content (living bacteria count) reach 2.03 × 1010cfu/ml。
Description
Technical field
The present invention relates to a kind of photosynthetic bacteria microbial inoculum and preparation method and application in microorganism field.
Background technology
Photosynthetic bacteria (Photosynthetic Bacteria, be called for short PSB) is that the comparison that nature is widely present most is ancient
, photosynthesis can be carried out and the general name of the prokaryotic micro-organisms of the special physiological monoid of non-oxygen-production, there is original luminous energy compound body
System, can not put the photosynthesis of oxygen, be primary producer main in water body amphimicrobian layer, and
The circulation of the carbon element of nature, nitrogen, sulphur transformations plays an important role.It includes Rhodospirillaceae
(Phodospirillaceae), Chromatiaceae (Chromati-aceae), Chlorobacteriaceae (Chlorobiaceae), green thread
Cordycepps (Chlo-roflexaceae) 4 section.
Rhodopseudomonas palustris is one of representative strain of photosynthetic bacteria, belongs to Rhodospirillaceae, Rhodopseudomonas, is me
The Ministry of Agriculture of state promulgates one of 12 kinds of living microorganisms allowing directly to add in feedstuff.According to country's agricultural microbial agent mark
Quasi-GB20287-2006, the living bacteria count standard of liquid photosynthetic bacteria microbial inoculum is >=2.0 hundred million/ml, shelf-life >=3 month.Training
Composition and the proportioning of supporting base all have important impact to reproduction speed and the preservation term length of Rhodopseudomonas palustris.Existing grind
Study carefully show Rhodopseudomonas palustris yeast extract exist under conditions of thalline breeding speed (Zhang Linghua, Kuang Zheshi, Chen Wei, contour
Activity photosynthetic bacteria Rhodopseudomonas palustris cultural character pre-test [J]. South China Normal University's journal (natural science edition), 2001
(4):37–391)。
Strain is the basis of microbial inoculum production application.At present, the bottleneck limiting the development of Rhodopseudomonas palustris microbial inoculum is exactly high
The effect selection-breeding problem of strain and the preparation of Rhodopseudomonas palustris microbial inoculum and preservation problem.
Summary of the invention
The technical problem to be solved is to provide a kind of preparation cost low (strain growth speed is fast), shelf-life
Grow, chicken production performance photosynthetic bacteria microbial inoculum that can improve chicken immuological function and chicken meat quality again and preparation method thereof can be improved.
In order to solve above technical problem, the invention provides a kind of photosynthetic bacteria microbial inoculum.
The active component of photosynthetic bacteria microbial inoculum provided by the present invention is Rhodopseudomonas palustris (Rhodopseudomonas
Palustris), the bacterial strain number of described Rhodopseudomonas palustris (Rhodopseudomonas palustris) is 981, its in
The numbered CGMCC No.10802 that registers on the books at state's Microbiological Culture Collection administration committee common micro-organisms center, below letter
Claim Rhodopseudomonas palustris 981.
Described photosynthetic bacteria microbial inoculum specifically can be made up of Rhodopseudomonas palustris 981 and carrier；Described carrier is concretely
At least one in powdered rice hulls, defatted milk powder, maltodextrin and wheat bran.
The preparation method of photosynthetic bacteria microbial inoculum provided by the present invention, including by Rhodopseudomonas palustris 981 in microorganism
Cultivating in culture medium and obtain Rhodopseudomonas palustris 981, the Rhodopseudomonas palustris 981 cultivation obtained is as photosynthetic bacteria bacterium
The active component of agent, obtains photosynthetic bacteria microbial inoculum.
In above-mentioned preparation method, described microbiological culture media can be fermentation medium B, described fermentation medium B by water and
Solute forms, and described solute is made up of bean sprout juice and NaCl, and described bean sprout juice is to extract, with water, the water solublity obtained from bean sprout
Material；In described fermentation medium B, the content of NaCl is 0.02-0.1g/L (such as 0.05g/L), and the content of bean sprout juice is in terms of bean sprout
It is 50-150g/L (such as 100g/L).
In above-mentioned preparation method, it can be to be boiled 30 minutes bean sprout with water that described water extracts from bean sprout.
In above-mentioned preparation method, described water can be distilled water or deionized water.
In above-mentioned preparation method, described bean sprout can be Semen Glycines Germinatus.
The preparation method of photosynthetic bacteria microbial inoculum provided by the present invention, including by Rhodopseudomonas palustris 981 at described
Ferment culture medium B is cultivated, collects the material in culture vessel, obtain described photosynthetic bacteria microbial inoculum.
Above, described cultivation can be in the intensity of illumination of 20-37 DEG C of (such as 28-37 DEG C or 30 DEG C) 24 hours every day
3 days-10 days (such as 3-7 days or 3 days) is cultivated under the illumination condition of 1000-1500Lx (1200-1500Lx or 1500Lx).
The photosynthetic bacteria microbial inoculum prepared by any of the above-described kind of preparation method falls within protection scope of the present invention.
Animal feed containing above-mentioned photosynthetic bacteria microbial inoculum falls within protection scope of the present invention.
In the above-mentioned animal feed containing photosynthetic bacteria microbial inoculum, the described animal feed containing photosynthetic bacteria microbial inoculum is general by chicken
Logical feedstuff and described photosynthetic bacteria microbial inoculum composition；Described chicken normal diet does not contains described photosynthetic bacteria microbial inoculum.
In the above-mentioned animal feed containing photosynthetic bacteria microbial inoculum, joining of described photosynthetic bacteria microbial inoculum and described chicken normal diet
Than meeting 2.5 × 107-5×107Chicken normal diet described in cfu (colony-forming units) Rhodopseudomonas palustris 981:100g.
In the above-mentioned animal feed containing photosynthetic bacteria microbial inoculum, described photosynthetic bacteria microbial inoculum and the tool of described chicken normal diet
Body proportioning meets 2.5 × 107Chicken normal diet described in cfu Rhodopseudomonas palustris 981:100g.
In the above-mentioned animal feed containing photosynthetic bacteria microbial inoculum, described chicken normal diet is except not containing above-mentioned photosynthetic bacteria bacterium
Outside agent, other raw material all can be identical with chicken conventional feed, as containing energy feed, protein feeds, mineral feed and Wei Sheng
Element feedstuff；Further, described chicken normal diet also can contain antioxidant.Described chicken normal diet can be by energy feed, albumen
Matter feedstuff, mineral feed and vitamin feed composition, it is possible to by energy feed, protein feeds, mineral feed, vitamin
Feedstuff and antioxidant composition, it is, of course, also possible to add preservative, coloring agent, flavoring agent, medicine in described chicken normal diet
The non-nutritive additives such as thing health-care agent, growth promoter.Described energy feed can be conventional chicken energy feed, as being selected from
At least one in following substances: Gu Shi, furfur, potato, grass seed and tree are real.Described protein feeds can be conventional egg albumen
Matter feedstuff, as being selected from least one in following substances: soybean meal, cottonseed meal, corn germ cake, meat meal tankage, rapeseed meal, flower
The raw dregs of rice and the certain herbaceous plants with big flowers dregs of rice etc..Described mineral feed is selected from stone powder, calcium bicarbonate, Sal, conch meal, bone meal, calcium hydrogen phosphate, carbonic acid
Calcium, Sal, copper sulfate, ferrous sulfate, manganese sulfate, potassium iodide, zinc oxide, sodium selenite and cobaltous chloride etc..Described vitamin is raised
Material can be conventional vitamin feed, as being selected from least one in following substances: vitamin A, vitamin D3, vitamin E,
Vitamin K3, vitamin B1 g, vitamin B2, vitamin B6, vitamin B12, nicotinic acid, D-VB5 calcium, biotin, choline chloride
Described antioxidant can be vitamin E (alpha-tocopherol) or Butylated hydroxyanisole (BHA).
Above-mentioned photosynthetic bacteria microbial inoculum, above-mentioned preparation method or above-mentioned animal feed preparation improve chicken immune performance and/or
The application improved in the product of chicken production performance and/or raising chicken meat quality falls within protection scope of the present invention.
Described product can be microbial ecological agent or animal feed.
Above-mentioned photosynthetic bacteria microbial inoculum, above-mentioned preparation method or above-mentioned animal feed are improving chicken production performance and/or raising
Chicken meat quality and/or the application improved in chicken immune performance fall within protection scope of the present invention.
Above-mentioned raising chicken immune performance can be following I1)-I5) in all or part of:
I1) broiler thymus index is improved；
I2) broiler index and spleen index is improved；
I3) broiler bursal index is improved；
I4) broiler erythrocyte C is improved3bReceptor rosette rate；
I5) broiler red blood cell immune complexes rosett is improved；
Described production performance is following P1)-P6) in all or part of:
P1) rate of animals delivered to the slaughter-house；
P2) meat feed ratio；
P3) apparent digestibility of protein；
P4) apparent digestibility of calcium；
P5) apparent digestibility of phosphorus；
P6) body weight；
Described raising chicken meat quality is following Q1)-Q8) in all or part of:
Q1) valine content of Carnis Gallus domesticus is improved；
Q2) methionine content of Carnis Gallus domesticus is improved；
Q3) phenylalanine content of Carnis Gallus domesticus is improved；
Q4) cystine of Carnis Gallus domesticus is improved；
Q5) tyrosine content of Carnis Gallus domesticus is improved；
Q6) crude protein content of Carnis Gallus domesticus is improved；
Q7) cholesterol level of Carnis Gallus domesticus is reduced；
Q8) fat content of Carnis Gallus domesticus is reduced.
Present invention also offers a kind of raising chicken production performance and/or improve chicken meat quality and/or improve chicken immune performance
Method.
Provided by the present invention improve chicken production performance and/or improve chicken meat quality and/or improve chicken immune performance side
Method, including with containing photosynthetic bacteria microbial inoculum Diet chicken；Described containing photosynthetic bacteria microbial inoculum daily ration by the common daily ration of chicken and described light
Close antibacterial microbial inoculum composition；The common daily ration of described chicken does not contains described photosynthetic bacteria microbial inoculum.
Described containing in photosynthetic bacteria microbial inoculum daily ration, the proportioning of described photosynthetic bacteria microbial inoculum and the common daily ration of described chicken is satisfied
2.5×107-5×107Cfu (colony-forming units) described Rhodopseudomonas palustris (Rhodopseudomonas
Palustris): the common daily ration of chicken described in 100g.
Above, described animal can be chicken.Described chicken can be broiler, such as AA broiler chicken or Arbor Acre chickens.
Above, described chicken can be chickling.
Active component Rhodopseudomonas palustris 981 reproduction speed of photosynthetic bacteria microbial inoculum of the present invention is fast, cultivates at bean sprout juice
In base (fermentation medium B) 30 DEG C, cultivate 3 days thalline under the illumination condition that intensity of illumination is 1500Lx of 24 hours every days and contain
Amount (living bacteria count) reaches 2.03 × 1010Cfu/ml, in containing yeast extract medium (fermentation medium A) 30 DEG C, every
It intensity of illumination of 24 hours be 1500Lx illumination condition under cultivate 3 days thalline content (living bacteria count) reach 8.37 ×
109cfu/ml.Photosynthetic bacteria microbial inoculum of the present invention in bean sprouts medium (fermentation medium B) at 20 DEG C 24 hours every days
Intensity of illumination is to reach 120 days the illumination condition lower shelf-life of 500Lx, photosynthetic bacteria microbial inoculum preservation 120 under this condition of the present invention
After it, the content of Rhodopseudomonas palustris 981 is 26.625 × 109Cfu/ml, is 1.31 times of preservation 0 day.
Photosynthetic bacteria microbial inoculum of the present invention can improve following I1)-I5) these 5 kinds of chicken immune performances, following Q1 can be improved again)-
Q8) these 8 kinds of chicken meat qualities, can improve again following P1)-P6) these 6 kinds of production performance: I1) improve broiler thymus index；I2) improve
Broiler index and spleen index；I3) broiler bursal index is improved；I4) broiler erythrocyte C is improved3bReceptor rosette rate；I5) broiler is improved
Red blood cell immune complexes rosett；P1) rate of animals delivered to the slaughter-house；P2) meat feed ratio；P3) apparent digestibility of protein；P4) calcium is apparent
Digestibility；P5) apparent digestibility of phosphorus；P6) body weight；Q1) valine content of Carnis Gallus domesticus is improved；Q2) methionine of Carnis Gallus domesticus is improved
Content；Q3) phenylalanine content of Carnis Gallus domesticus is improved；Q4) cystine of Carnis Gallus domesticus is improved；Q5) tyrosine improving Carnis Gallus domesticus contains
Amount；Q6) crude protein content of Carnis Gallus domesticus is improved；Q7) cholesterol level of Carnis Gallus domesticus is reduced；Q8) fat content of Carnis Gallus domesticus is reduced.
Preservation explanation
Strain name: Rhodopseudomonas palustris (Rhodopseudomonas palustris)
Strain number: 981
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on May 11st, 2015
Register on the books numbering in preservation center: CGMCC No.10802
Detailed description of the invention
Being further described in detail the present invention below in conjunction with detailed description of the invention, the embodiment be given is only for explaining
The bright present invention rather than in order to limit the scope of the present invention.Experimental technique in following embodiment, if no special instructions, is
Conventional method.Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
Embodiment 1, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Isolation identification
1.1 strains separation
Water sample is gathered from a pond of China Wuxi City, Jiangsu Province.Use the enrichment training of vanNeil purple nonsulfur bacteria
Foster base carries out enrichment culture, successively enrichment three times, until photosynthetic bacteria becomes dominant microflora in culture medium.By light later for enrichment
Close bacteria samples and separate the single bacterium colony of line separation, illumination cultivation 24 in 28 DEG C of anaerobic jars on solid medium photosynthetic bacteria respectively
Hour.Single bacterium colony on flat board is chosen in liquid photosynthetic bacteria isolation medium continuation cultivate.And repeat above step,
Obtain the bacterial strain of purification.The bacterial strain (bacterial strain 981) taking numbered 981 carries out following qualification.
Wherein, the compound method of vanNeil purple nonsulfur bacteria enrichment medium is as follows:
NH4Cl 0.1g, NaCl 0.2g, NaH2P040.05g, MgCl20.02g, yeast extract 0.15g, by above each group
Divide and be dissolved in 100ml water, 121 DEG C of steam sterilization 20min, make basal medium.This basal medium added and filtered
The 10%NaHCO of bacterium3Aqueous solution 50ml, the ethanol 1.5ml of filtration sterilization, adjust pH value 7.0 with the phosphoric acid of filtration sterilization.
The compound method of photosynthetic bacteria isolation medium is as follows:
Sodium acetate 3.0g, sodium propionate 0.3g, NaCl 1.0g, (NH4)2SO40.3g, MgSO40.2g, KH2P040.5g,
K2HSO40.3g, CaC1250mg, MnSO42.5mg, FeSO45mg, yeast extract 0.1g, peptone 10mg, glutamic acid 0.2mg,
Water 1000ml, pH7.4.PH, then 121 DEG C of steam sterilization 30min is adjusted after above component is dissolved in water, if configuration solid culture
Base then adds 17g agar steam sterilization again in above composition, and pour plate, for separating single bacterium colony after doing Sterility testing.
1.2 identification of strains
Bacterial strain 981 thalline is shaft-like, occasionally has micro-bend, and size is 0.5-0.8 × 1.0-2.5um, the light of the raw single flagellar movement of end
Can heterotrophic bacteria.Gram-negative, photosynthetic endomembrane system is sheet.Under illumination anaerobic condition, culture is peony, dark good
Oxygen is then light red, and aged bacterium is gathered into rose-shaped.Para-amino benzoic acid is essential growth factor.This bacterium has Bacteriochlorin a
The class carotene of ether system purplish red with standard, can utilize formates, acetate, acetone acid, lactate, succinate, glycerol,
The small organic molecules such as aspartic acid, benzoate, sodium thiosulfate and hydrogen are photosynthetic electron donor, it is impossible to profit
With monosaccharide, sugar alcohol and sulfide.Growth pH scope 5.5-8.5, optimum pH 6.8-7.0.Growth temperature range 20-37 DEG C, the suitableeest
Temperature range 28-37 DEG C.
The 16S rRNA gene order of bacterial strain 981 is as shown in the sequence 1 in sequence table, with Rhodopseudomonas palustris
The homology of (Rhodopseudomonas palustris) is the highest, reaches 99%.
Therefore, comprehensive features above determines that bacterial strain 981 is for Rhodopseudomonas palustris (Rhodopseudomonas
palustris)。
Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 was preserved on 05 11st, 2015
China General Microbiological culture presevation administration committee's common micro-organisms center, deposit number is CGMCC No.10802.
Embodiment 2, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Cultivate
The present embodiment uses fermentation medium A and fermentation medium B both culture medium culturing Rhodopseudomonas palustris
(Rhodopseudomonas palustris)981CGMCC No.10802.Fermentation medium A is presently considered to be and is best suitable for marsh
The culture medium containing yeast extract of rhodopseudomonas, culture medium B is the culture medium that the present inventor screens.
The compound method of fermentation medium A is as follows: by tryptone 3.0g, yeast leaching powder 3.0g, CaCl20.3g and
MgSO4·7H2After O 0.5g is dissolved in distilled water, being settled to 1000mL with distilled water, then 121 DEG C of steam sterilization 30min, obtain
Culture medium A.
The compound method of fermentation medium B is as follows: boiled 30 minutes by 100g (fresh weight) Semen Glycines Germinatus distilled water, filters and receives
Collecting whole filtrate, this filtrate is bean sprout juice, adds 0.05g NaCl, be settled to distilled water in whole bean sprout juices
1000mL, then 121 DEG C of steam sterilization 30min, obtain culture medium B.
Wherein, Semen Glycines Germinatus is made up of radicle, plumule, plumular axis and cotyledon, and radicle length is 1-2cm.
1, the activation of strain
Picking Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802, to it
Carrying out inclined-plane stab culture with preferred solid medium, keeping 30 DEG C of intensities of illumination of temperature is cultivation 10d under the conditions of 1200Lx.Excellent
Selecting solid medium is to add the solid medium that agar obtains in fermentation medium A, and concrete compound method is as follows: by pancreas egg
White peptone 3.0g, yeast leaching powder 3.0g, CaCl2 0.3g、MgSO4·7H2After O 0.5g and 17g agar are dissolved in distilled water, with distillation
Water is settled to 1000mL, then 121 DEG C of steam sterilization 30min, obtains preferred solid medium.
2, preparation seed liquor
Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC that step 1 is activated
No.10802 is inoculated in the aseptic blue lid reagent bottle (Clear glass bottles and jars of screw thread mouth) of the fermentation medium A equipped with embodiment 2,
Tighten blue lid, 30 DEG C, cultivate 5 days under the illumination condition that intensity of illumination is 1200Lx of 24 hours every days, obtain the red vacation in marsh
Zymomonas mobilis (Rhodopseudomonas palustris) 981CGMCC No.10802 seed liquor.
3, fermentation culture
3.1 utilize fermentation medium B to cultivate Rhodopseudomonas palustris (Rhodopseudomonas palustris)
981CGMCC No.10802
By Rhodopseudomonas palustris (Rhodopseudomonas palustris) the 981CGMCC No.10802 of step 2
Seed liquor 10ml accesses the aseptic blue lid reagent bottle (Clear glass bottles and jars of screw thread mouth) of the 100ml equipped with 100ml fermentation medium B
In, tighten blue lid, 30 DEG C, cultivate 3 days under the illumination condition that intensity of illumination is 1500Lx of 24 hours every days, obtain marsh red
The fermentation liquid (being called for short fermentation liquid B) of the fermentation medium B of pseudomonas (Rhodopseudomonas palustris) 981.Real
Test in triplicate, repeat 10 blue lid reagent bottle every time.
3.2 utilize fermentation medium A to cultivate Rhodopseudomonas palustris (Rhodopseudomonas palustris)
981CGMCC No.10802
This step from 3.1 to differ only in fermentation medium used different, other condition is the most identical, and concrete grammar is such as
Under:
By Rhodopseudomonas palustris (Rhodopseudomonas palustris) the 981CGMCC No.10802 of step 2
Seed liquor 10ml accesses the aseptic blue lid reagent bottle (Clear glass bottles and jars of screw thread mouth) of the 100ml equipped with 100ml fermentation medium A
In, tighten blue lid, 30 DEG C, cultivate 3 days under the illumination condition that intensity of illumination is 1500Lx of 24 hours every days, obtain marsh red
The fermentation liquid (being called for short fermentation liquid A) of the fermentation medium A of pseudomonas (Rhodopseudomonas palustris) 981.Real
Test in triplicate, repeat 10 blue lid reagent bottle every time.
The fermentation medium B of the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 to 3.1 sends out
Ferment liquid and the fermentation liquid of fermentation medium A of Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 of 3.2
Count according to plating dilutions counting method: measure 10ml (being accurate to 0.01ml) fermentation liquid, put into built-in 90ml sterilized water
In triangular flask, put shaking table vibration 30min, rotating speed 200r/min.Stand 20min after vibration and make the uniform diluent of 1:10.Adopt
With 10 times of dilution method dilutions.
Draw 1:10 diluent 5ml with sterilizing suction pipe, inject in the triangular flask containing 45ml sterilized water, make 1:100's
Diluent, makes 10 times of incremental diluents successively.
Choose three appropriate dilution suspensions and respectively take 0.1ml, add in the culture dish of a diameter of 90mm, pour 45 DEG C into preferably
Solid medium, mixes gently, to be solidified.Each dilution factor is in triplicate.Keeping 30 DEG C of intensities of illumination of temperature is 1200Lx bar
10d is cultivated under part.
Calculating bacterium number: thalline content (cfu/ml)=bacterium colony average × extension rate × basal liquid volume/(sample size ×
Sample-adding amount).
Result shows, the fermentation medium A's of Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981
Thalline content average out to 8.37 × 10 in fermentation liquid9Cfu/ml, Rhodopseudomonas palustris (Rhodopseudomonas
Palustris) the thalline content average out to 2.03 × 10 in the fermentation liquid of the fermentation medium B of 98110Cfu/ml, the red vacation in marsh
Thalline content in the fermentation liquid of the fermentation medium B of Zymomonas mobilis (Rhodopseudomonas palustris) 981 is marsh
Thalline content in the fermentation liquid of the fermentation medium A of rhodopseudomonas (Rhodopseudomonas palustris) 981
2.42 again.
Table 1, the thalline content of different fermentations liquid
Embodiment 3, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802 increase
Strong broiler immunologic function and body weight
1, material
1.1, the preparation of Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 microbial inoculums
Fermentation medium B to the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 of embodiment 2
Fermentation liquid in add powdered rice hulls be dried at 50 DEG C, obtain Rhodopseudomonas palustris (Rhodopseudomonas
Palustris) 981 microbial inoculum.Marsh in this Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 microbial inoculums
Rhodopseudomonas (Rhodopseudomonas palustris) 981 content are 1 × 109cfu/g.Wherein, the prior warp of powdered rice hulls
(powder granularity of 90% is less than 100 μm, and mean diameter is less than 50 μm, and described mean diameter is 50% powder body to cross micronization processes
Particle diameter).
1.2, experimental animal
The love selecting body weight to be 50 ± 5 grams pulls out the strong young bird of 1 age in days of the fastest large-scale white meat-type chickens, is divided into 3 at random
Process group, i.e. matched group, microbial inoculum I group and microbial inoculum II group.Each process group 3 repetition, 300 chickens of each repetition.
1.3, daily ration
The formula of the common daily ration of broiler in following embodiment is as shown in table 2.
Table 2, broiler common daily ration composition and proportioning
In table 2, every kilogram of premix material contains: CuSO4·5H2O 0.64g,FeSO4·H2O 516g,ZnSO4·H2O 4g,
MnSO4·H2O 318g,Na2SeO30.016g,VA16IU,VD3 415IU,VE 400IU,VK 90mg,VB1 60mg,
VB2200mg, nicotinic acid 550mg, calcium pantothenate 220mg, folic acid 12mg, remaining is stone powder.
" % " in table 2 is weight/mass percentage composition.
Broiler brood time daily ration is divided into the common daily ration of broiler brood time, I group of brood time daily ration of microbial inoculum and microbial inoculum II group to brood
Phase daily ration.Wherein, the common daily ration of broiler brood time is as shown in table 2；I group of brood time daily ration of microbial inoculum is the broiler brood time at table 2
In common daily ration, interpolation is the Rhodopseudomonas palustris of the 0.025% of the broiler brood time common daily ration quality of table 2
The daily ration that (Rhodopseudomonas palustris) 981 microbial inoculums obtain；II group of brood time daily ration of microbial inoculum is the broiler at table 2
In the common daily ration of brood time, interpolation is the Rhodopseudomonas palustris of the 0.05% of the broiler brood time common daily ration quality of table 2
The daily ration that (Rhodopseudomonas palustris) 981 microbial inoculums obtain.
Broiler finishing period daily ration is divided into the common daily ration of broiler finishing period, I group of finishing period daily ration of microbial inoculum and microbial inoculum II group incubation
Phase daily ration.Wherein, the common daily ration of broiler finishing period is as shown in table 2；I group of finishing period daily ration of microbial inoculum is the broiler finishing period at table 2
In common daily ration, interpolation is the Rhodopseudomonas palustris of the 0.025% of the broiler finishing period common daily ration quality of table 2
The daily ration that (Rhodopseudomonas palustris) 981 microbial inoculums obtain；II group of finishing period daily ration of microbial inoculum is the broiler at table 2
In the common daily ration of finishing period, interpolation is the Rhodopseudomonas palustris of the 0.05% of the broiler finishing period common daily ration quality of table 2
The daily ration that (Rhodopseudomonas palustris) 981 microbial inoculums obtain.
2, meat is improved with Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Immune Organs of Chicken index and hematid immunity function
28 days experimental periods, all broiler free choice feedings and drinking-water during test, carry out immunity according to routine immunization program, respectively
Group is in addition to the daily ration difference fed, and other rearing conditions is identical.Wherein, matched group is at the 1-28 age in days broiler of table 2
The common daily ration of brood time feeds；Microbial inoculum I group feeds at I group of brood time daily ration of 1-28 age in days microbial inoculum；Microbial inoculum II
Group feeds at II group of brood time daily ration of 1-28 age in days microbial inoculum.
The spleen of broiler, thymus and bursal index is measured respectively at 14 ages in days, 21 ages in days, 28 ages in days.Assay method is:
Each repeating to randomly draw 20 chickens, weigh in the morning on an empty stomach, wins spleen, fabricius bursa and both sides thymus after butchering, and rejects fat
With scales/electronic balance weighing after fat, Computation immunity shoot formation.Immune Organs Index=immune organ weight (mg)/body weight (g).According to
Following method measures 21 Day-old Broiler Chickens erythrocyte C3bReceptor garland (E-C3bR) rate and Erythrocyte immune complex rosette rate (E-ICR)
Rate: often organize and each repeat to randomly draw 6 chickens, anticoagulant of taking a blood sample, centrifuging and taking erythrocyte, it is made into 1.25 × 107The erythrocyte of individual/mL
Suspension, as red cell suspension to be measured.By yeast with after brine 3 times, it is made into 1% suspension, boils in putting water-bath
20min, fully mixes.Filter with two layers of qualitative filter paper, remove little grumeleuse, in single yeast cell dispersed under low power lens
State.Add equivalent mouse serum, mix rearmounted 37 DEG C of water-bath sensitization 15min.With brine once, 2500r/min from
Heart 10min.Remove most supernatant, with normal saline weight suspendible.1 × 10 it is made into after counting8The yeast of/ml complement (C3) sensitization hangs
Liquid.It is measured in accordance with the following steps:
(1) taking two small test tubes, often pipe adds red cell suspension 100ul to be measured, and the first pipe adds complement Sensitized yeast again and hangs
Liquid 100ul, the second pipe adds unsensitized yeast suspension 100ul, shakes up and put 37 DEG C of water-bath 30min；
(2) taking-up wrist shakes up gently, adds normal saline 200ul；
(3) after mixing, then add 0.25% glutaraldehyde 75 μ l and mix gently；
(4) take the 1/3 horizontal smear of amount respectively, dry up, add methanol and fix；
(5) dye by Rui Shi method, count under oil mirror.After smear staining, RBC is red, and yeast is blue.On erythrocyte
Adhere to 2 or more than 2 yeast persons are garland.200 erythrocyte of counting, calculate garland positive percentage respectively.The
One pipe is RBC-C3b receptor rosette rate, and the second pipe is RBC-IC rosette rate.
Garland forms percentage rate (%)=(red blood cell rosette number/200) × 100.
The independent samples t test testing all data acquisition SPSS12.0 (SPSS Inc., USA) statistical software processes system
Meter.
Result is as shown in table 3, and when 14 ages in days and 21 age in days, the index and spleen index of microbial inoculum I group and microbial inoculum II group is the most significantly high
In matched group (P < 0.01)；During 14 age in days, thymus index all pole of microbial inoculum I group and microbial inoculum II group be significantly higher than matched group (P <
0.01)；Thymus index, index and spleen index and the bursal index of microbial inoculum I group and II group of other age in days of microbial inoculum are all remarkably higher than comparison
Group (P < 0.05).Show Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 pairs of broiler spleens, fabricius bursa
Growth with thymus has facilitation.
Table 3, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 pairs of broiler Immune Organs Indexes
Impact (mg/g)
Note: the different lower case person's significant difference (P < 0.05) of colleague's data mark, the different capitalization person's difference of mark
Extremely notable (P < 0.01).
As shown in table 4, microbial inoculum I group and the (E-C of microbial inoculum II group3bR) rate and (E-ICR) rate all pole are significantly higher than matched group (P
< 0.01), and (the E-C of microbial inoculum I group3bR) rate and (E-ICR) rate all pole are significantly higher than microbial inoculum II group.Show the red false unit cell in marsh
981 pairs of chicken red blood cell immunologic functions of bacterium (Rhodopseudomonas palustris) have facilitation.
Table 4, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 are to 21 Day-old Broiler Chickens erythrocyte flowers
The measurement result (%) of ring rate
Note: with the different lower case person's significant difference (P < 0.05) of column data mark, the different capitalization person's difference of mark
Extremely notable (P < 0.01).
3, meat is improved with Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Chicken body weight
42 days experimental periods, all broiler free choice feedings and drinking-water during test, carry out immunity according to routine immunization program, respectively
Group is in addition to the daily ration difference fed, and other rearing conditions is identical.Wherein, matched group is at the 1-28 age in days broiler of table 2
The common daily ration of brood time feeds, and the common daily ration of broiler finishing period at 29-42 age in days table 2 feeds；Microbial inoculum I group
Feed at I group of brood time daily ration of 1-28 age in days microbial inoculum, carry out at I group of finishing period daily ration of 29-42 age in days microbial inoculum
Feed；Microbial inoculum II group feeds at II group of brood time daily ration of 1-28 age in days microbial inoculum, at 29-42 age in days microbial inoculum II group
Finishing period daily ration feeds.
The body weight of broiler during determination test.Assay method is: respectively at 7 ages in days, 14 ages in days, 21 ages in days, 28 ages in days, 42
Age in days is weighed once respectively, and fasting in early morning is prohibited water and weighed.When weighing, each repetition randomly draws 100 chickens, respectively records.
The independent samples t test testing all data acquisition SPSS12.0 (SPSS Inc., USA) statistical software processes system
Meter.
Result shows, during 7 age in days, each group broiler weight difference is not notable (P ＞ 0.05)；During 14 ages in days to 42 age in days, bacterium
The broiler live-weight of agent I group and microbial inoculum II group is all remarkably higher than matched group (P < 0.05).Show Rhodopseudomonas palustris
(Rhodopseudomonas palustris) 981 has good facilitation to improving broiler live-weight.
Table 5, the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 impact (g) on broiler live-weight
| Age in days (d) | Matched group | Microbial inoculum I group | Microbial inoculum II group |
| 7 | 196.2±3.84ab | 207.6±4.08a | 184.3±4.15b |
| 14 | 312.5±6.53b | 338.7±6.25a | 332.1±7.99a |
| 21 | 718.6±16.28b | 759.5±15.10a | 751.3±14.52a |
| 28 | 1168.1±18.77b | 1295.9±14.28a | 1265.2±16.22a |
| 35 | 1560.9±18.79b | 1714.2±21.35a | 1703.3±19.01a |
| 42 | 1898.9±21.02b | 2095.8±22.21a | 2085.2±20.12a |
Note: the different significance degree of the English alphabet differential of colleague in table, in 0.05 level without aobvious between the process of same letter
Writing difference, between the process that letter is different, in 0.05 level, there were significant differences.
Embodiment 4, use Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Improve meat chicken production performance and improve Meat Quality of Broiler Chicks
1, material
1.1, the preparation of Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 microbial inoculums
Fermentation medium B to the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 of embodiment 2
Fermentation liquid in add powdered rice hulls be dried at 50 DEG C, obtain Rhodopseudomonas palustris (Rhodopseudomonas
Palustris) 981 microbial inoculum.Marsh in this Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 microbial inoculums
Rhodopseudomonas (Rhodopseudomonas palustris) 981 content are 1 × 109cfu/g.Wherein, the prior warp of powdered rice hulls
(powder granularity of 90% is less than 100 μm, and mean diameter is less than 50 μm, and described mean diameter is 50% powder body to cross micronization processes
Particle diameter).
1.2, experimental animal
The strong young bird of 1 age in days selecting body weight to be 50 ± 5 Ke Aiwei mattress white meat-type chickens, is divided into 3 process groups at random, the most right
According to group, microbial inoculum I group and microbial inoculum II group.Each process group 3 repetition, 300 chickens of each repetition.
1.3, daily ration
With embodiment 3 1.3.
2, meat is improved with Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Chicken production performance
47 days experimental periods, all broiler free choice feedings and drinking-water during test, carry out immunity according to routine immunization program, respectively
Group is in addition to the daily ration difference fed, and other rearing conditions is identical.Wherein, matched group is at the 1-28 age in days broiler of table 2
The common daily ration of brood time feeds, and the common daily ration of broiler finishing period at 29-47 age in days table 2 feeds；Microbial inoculum I group
Feed at I group of brood time daily ration of 1-28 age in days microbial inoculum, carry out at I group of finishing period daily ration of 29-47 age in days microbial inoculum
Feed；Microbial inoculum II group feeds at II group of brood time daily ration of 1-28 age in days microbial inoculum, at 29-47 age in days microbial inoculum II group
Finishing period daily ration feeds.The mortality rate of broiler, the rate of animals delivered to the slaughter-house, feed intake and feedstuff-meat ratio during determination test.On 36th
Age gathers broiler excrement sample and daily ration sample to 47 ages in days, measures broiler with endogenous indicator method (4mol/L ash insoluble in hydrochloric acid)
Nutrient substance apparent digestibility.Crude protein (CP) content in daily ration and excrement sample uses crude protein in GB/T 6432-1994 feedstuff
Assay method, calcium content uses the mensuration (computed microstructure) of NY/T1944-2010 Calcium in Feed, and phosphorus content uses
Total phosphorus yield (spectrophotography) in GB/T 6437-2002 feedstuff.
Computing formula is as follows:
Mortality rate (%)=death chicken number (only)/enter to give up chicken number (only) × 100；
The rate of animals delivered to the slaughter-house (%)=deliver for sale chicken number (only)/enter to give up chicken number (only) × 100；
Average daily ingestion amount=(dispensing total amount remains doses)/test natural law * often organizes chicken number
Average daily gain=(test end average weight test just average weight)/test natural law
Feedstuff-meat ratio=average daily ingestion amount/average daily gain.
Diet nutrient material apparent digestibility (%)=[1-(b/a) × (c/d)] × 100.Wherein, certain battalion during a is daily ration
The content (%) formed point；B is the content (%) of certain nutritional labeling in feces；C is the content of ash insoluble in hydrochloric acid in daily ration
(%)；D is the content (%) of ash insoluble in hydrochloric acid in feces.
The independent samples t test testing all data acquisition SPSS12.0 (SPSS Inc., USA) statistical software processes system
Meter.
Result shows, the feedstuff-meat ratio that microbial inoculum is I group is substantially less than matched group (P < 0.05) (table 6).Show the red false unit cell in marsh
Bacterium (Rhodopseudomonas palustris) 981 can reduce broiler feedstuff-meat ratio.
Table 6, the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 impact on broiler feedstuff-meat ratio
| Process | Feedstuff-meat ratio |
| Matched group | 1.98a |
| Microbial inoculum I group | 1.83b |
| Microbial inoculum II group | 1.94ab |
Note: the different letter of same column mark is significant difference (P < 0.05).
Result shows, the mortality rate of microbial inoculum I group and microbial inoculum II group all be substantially less than matched group (P < 0.05), microbial inoculum I group and
The rate of animals delivered to the slaughter-house that microbial inoculum is II group is all remarkably higher than matched group (P < 0.05) (table 7).Show Rhodopseudomonas palustris
(Rhodopseudomonas palustris) 981 can reduce broiler mortality rate, increases the number of heads of livestock for sale.
Table 7, the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 impact on broiler mortality rate
Note: the different letter of same column mark is significant difference (P < 0.05).
Result shows, crude protein (CP) apparent digestibility of microbial inoculum I group and microbial inoculum II group, calcium (Ca) apparent digestibility, phosphorus
(P) apparent digestibility is all remarkably higher than matched group (P < 0.05) (table 8).Rhodopseudomonas palustris is described
(Rhodopseudomonas palustris) 981 can improve broiler apparent digestibility of crude protein, calcium apparent digestibility and phosphorus table
See digestibility.
Table 8,981 pairs of daily ration of broiler apparent digestibilities of Rhodopseudomonas palustris (Rhodopseudomonas palustris)
Impact
Note: the different letter of same column mark is significant difference (P < 0.05).
3, meat is improved with Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Chicken muscle quality
47 days experimental periods, all broiler free choice feedings and drinking-water during test, carry out immunity according to routine immunization program, respectively
Group is in addition to the daily ration difference fed, and other rearing conditions is identical.Wherein, matched group is at the 1-28 age in days broiler of table 2
The common daily ration of brood time feeds, and the common daily ration of broiler finishing period at 29-47 age in days table 2 feeds；Microbial inoculum I group
Feed at I group of brood time daily ration of 1-28 age in days microbial inoculum, carry out at I group of finishing period daily ration of 29-47 age in days microbial inoculum
Feed；Microbial inoculum II group feeds at II group of brood time daily ration of 1-28 age in days microbial inoculum, at 29-47 age in days microbial inoculum II group
Finishing period daily ration feeds.Experimental period, terminates the previous day, often repeats to choose the close chicken of body weight 3, and cervical region blood-letting is butchered, and kills
Front fasting 12h.Measure cholesterol level, crude protein content and amino acid content in chicken whole-body muscle.
3.1 cholesterol levels are measured according to GB/T 5009.128-2003 " mensuration of Food Cholesterol " standard.
3.2 gross protein values measure
Weigh meat sample, then smash to pieces with tissue mashing machine, put into baking oven constant temperature drying.In digestion tube add 2g meat sample,
2.5g antioxidant and 10mL concentrated sulphuric acid, then carry out treatments of the sample on High-temperature Digestion stove.Application kjeldahl apparatus is surveyed
Fixed, computing formula is:
Crude protein %=(A-A0) × 0.014 × 6.25 × 0.1612/W × 100%
A0For blank, W is meat sample weight (g), and 0.1612 is sulfuric acid concentration, and A is machine-readable number.
3.3 crude fat content measure
Take meat sample, then smash mixing to pieces with tissue mashing machine, in an oven constant temperature drying, in filtration paper cylinder, place 1g sample
Product；Dry filtration paper cylinder, until weighing is constant weight, Milko-Tester carries out the mensuration of crude fat.
3.4 Contents of Amino Acids
It is measured according to GB/T 5009.124-2003 " amino acid whose mensuration in food " standard
The independent samples t test testing all data acquisition SPSS12.0 (SPSS Inc., USA) statistical software processes system
Meter.
Result shows that the cholesterol level of the muscle of microbial inoculum I group and microbial inoculum II group and crude fat content are substantially less than comparison
Group, crude protein content is significantly higher than matched group (P < 0.05) (table 9).Rhodopseudomonas palustris (Rhodopseudomonas is described
Palustris) 981CGMCC No.10802 can improve the nutritional labeling of meat quality of table poultry effectively.
Table 9, the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 impact on nutritive composition in muscle
| Testing index | Matched group | Microbial inoculum I group | Microbial inoculum II group |
| Cholesterol (mg/100g) | 79.63±1.59a | 75.58±1.35b | 75.81±1.55b |
| Crude protein (%) | 24.71±0.26b | 26.72±0.21a | 26.39±0.27a |
| Crude fat (%) | 4.46±0.19a | 3.62±0.15b | 3.79±0.18b |
Note: the different letter of colleague's mark is significant difference (P < 0.05).
Result shows microbial inoculum I group and valine, methionine, phenylalanine, cystine and the tyrosine of II group of muscle of microbial inoculum
Content is significantly higher than matched group (P < 0.05), and other amino acid content is not significantly different from (table 10).The increase of valine content,
Alanine in blood plasma can be made to raise, and improve animal body immune function；The increase of methionine content, can improve body
Immunity, reduction Dietary Crude Protein matter level, improve production performance in certain degree simultaneously；The rising of lysine, permissible
Improve gastric secretion effect, play appetite stimulator, promote to grow and the effect of growth, improve absorbing of calcium and amassing in vivo thereof
Tired, accelerate bone growth.Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC is described
No.10802 can improve the valine of chicken muscle, methionine, phenylalanine, cystine and tyrosine content.
Table 10, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 pairs of Amino Acid Contents of Muscles
Impact
Note: the different letter of colleague's mark is significant difference (P < 0.05).
Embodiment 4, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802
Preserve
1, the activation of strain
Picking Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC No.10802, to it
Carrying out inclined-plane stab culture with preferred solid medium, keeping 30 DEG C of intensities of illumination of temperature is cultivation 10d under the conditions of 1200Lx.Excellent
Selecting solid medium is to add the solid medium that agar obtains in fermentation medium A, and concrete compound method is as follows: by pancreas egg
White peptone 3.0g, yeast leaching powder 3.0g, CaCl2 0.3g、MgSO4·H2After O 0.5g and 17g agar are dissolved in distilled water, use distilled water
It is settled to 1000mL, then 121 DEG C of steam sterilization 30min, obtains preferred solid medium.
2, preparation seed liquor
Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981CGMCC that step 1 is activated
No.10802 is inoculated in the aseptic blue lid reagent bottle (Clear glass bottles and jars of screw thread mouth) of the fermentation medium A equipped with embodiment 2,
Tighten blue lid, 30 DEG C, cultivate 5 days under the illumination condition that intensity of illumination is 1200Lx of 24 hours every days, obtain the red vacation in marsh
Zymomonas mobilis (Rhodopseudomonas palustris) 981CGMCC No.10802 seed liquor.
3, fermentation culture
By Rhodopseudomonas palustris (Rhodopseudomonas palustris) the 981CGMCC No.10802 of step 2
Seed liquor 10ml accesses equipped with the aseptic blue lid reagent bottle of the 100ml of the fermentation medium B of 100ml embodiment 2 (screw thread mouth transparent
Vial) in, tighten blue lid, 30 DEG C, cultivate 3 days under the illumination condition that intensity of illumination is 1500Lx of 24 hours every days,
Fermentation liquid to the fermentation medium B of Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 (is called for short fermentation
Liquid B), this fermentation liquid B is Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 liquid bacterial agents.Experiment
In triplicate, repeat 10 blue lid reagent bottle every time.
4, preservation
The fermentation liquid B of step 3 is carried out B1 and B2 both preservations:
B1, under the illumination condition that intensity of illumination is 500Lx, carry out preservation
Ferment the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 just terminating to obtain by step 3
Liquid bacterial agent proceeds in aseptic blue lid reagent bottle (Clear glass bottles and jars of screw thread mouth), by Rhodopseudomonas palustris
(Rhodopseudomonas palustris) 981 liquid bacterial agents are filled to bottleneck, tighten blue lid, are placed in 20 DEG C 24 hours every days
The illumination condition that intensity of illumination is 500Lx under place 30 days, 60 days, 90 days, 120 days, 150 days, 180 days and 210 days respectively
After, count according to the plating dilutions counting method of embodiment 2.Ferment the Rhodopseudomonas palustris just terminating to obtain by step 3
(Rhodopseudomonas palustris) 981 liquid bacterial agents are designated as placing 0 day.Each process in triplicate, repeats every time
10 blue lid reagent bottle.
B2, under the illumination condition that intensity of illumination is 10Lx, carry out preservation
This step and B1 to differ only in intensity of illumination different, other condition is the most identical, and concrete grammar is as follows:
Ferment the Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 just terminating to obtain by step 3
Liquid bacterial agent proceeds in aseptic blue lid reagent bottle (Clear glass bottles and jars of screw thread mouth), by Rhodopseudomonas palustris
(Rhodopseudomonas palustris) 981 liquid bacterial agents are filled to bottleneck, tighten blue lid, are placed in 20 DEG C 24 hours every days
The illumination condition that intensity of illumination is 10Lx under place 30 days, 60 days, 90 days, 120 days, 150 days, 180 days and 210 days respectively
After, count according to the plating dilutions counting method of embodiment 2.Ferment the Rhodopseudomonas palustris just terminating to obtain by step 3
(Rhodopseudomonas palustris) 981 liquid bacterial agents are designated as placing 0 day.Each process in triplicate, repeats every time
10 blue lid reagent bottle.
Result shows that Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 liquid bacterial agents are in B1 condition
After lower preservation 120 days, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 (is called for short the red false unit cell in marsh
Bacterium 981) content be 26.625 × 109Cfu/ml, is 1.31 times of preservation 0 day, Rhodopseudomonas palustris after preservation 150 days
The content of (Rhodopseudomonas palustris) 981 (being called for short Rhodopseudomonas palustris 981) is 12.375 × 109cfu/
Ml, is 0.61 times of preservation 0 day；Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 liquid bacterial agents exist
Under the conditions of B2, after preservation 60 days, Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 (is called for short the red vacation in marsh
Zymomonas mobilis 981) content be 12.0 × 109Cfu/ml, is preservation 0.59 times (table 11) of 0 day.Rhodopseudomonas palustris
(Rhodopseudomonas palustris) 981 liquid bacterial agents shelf-life under the conditions of B1 is 120 days, under the conditions of B2
Shelf-life is 60 days.Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 liquid bacterial agents are under the conditions of B1
Meet in agricultural microbial bacterial agent standard (GB20287-2006) and be more than or equal to 2.0 hundred million about liquid dosage form microbial inoculum living bacteria count
Cfu/ml, the shelf-life is more than or equal to trimestral regulation.
Rhodopseudomonas palustris (Rhodopseudomonas palustris) after table 11, B1 and B2 preservation different time
Rhodopseudomonas palustris (Rhodopseudomonas palustris) 981 content in 981 liquid bacterial agents
Claims (10)
1. photosynthetic bacteria microbial inoculum, it is characterised in that: the active component of described photosynthetic bacteria microbial inoculum is Rhodopseudomonas palustris
(Rhodopseudomonas palustris), described Rhodopseudomonas palustris (Rhodopseudomonas palustris)
Bacterial strain number is 981, and it is numbered in registering on the books of China Committee for Culture Collection of Microorganisms's common micro-organisms center
CGMCC No.10802。
2. the preparation method of photosynthetic bacteria microbial inoculum described in claim 1, including by Rhodopseudomonas palustris
(Rhodopseudomonas palustris) cultivates in microbiological culture media and obtains described Rhodopseudomonas palustris
(Rhodopseudomonas palustris), the Rhodopseudomonas palustris (Rhodopseudomonas that cultivation is obtained
Palustris) as the active component of photosynthetic bacteria microbial inoculum, photosynthetic bacteria microbial inoculum is obtained；Described Rhodopseudomonas palustris
The bacterial strain number of (Rhodopseudomonas palustris) is 981, and it is general in China Committee for Culture Collection of Microorganisms
The numbered CGMCC No.10802 that registers on the books at logical microorganism center.
Preparation method the most according to claim 2, it is characterised in that: described microbiological culture media is fermentation medium B, institute
Stating fermentation medium B to be made up of water and solute, described solute is made up of bean sprout juice and NaCl, and described bean sprout juice is from bean sprout with water
The water-soluble substances that middle extraction obtains；In described fermentation medium B, the content of NaCl is 0.02-0.1g/L, the content of bean sprout juice
It is 50-150g/L in terms of bean sprout.
4. the preparation method of photosynthetic bacteria microbial inoculum described in claim 1, it is characterised in that: described preparation method includes marsh red
Pseudomonas (Rhodopseudomonas palustris) is cultivated in fermentation medium B, collects the material in culture vessel,
Obtain described photosynthetic bacteria microbial inoculum；The bacterial strain number of described Rhodopseudomonas palustris (Rhodopseudomonas palustris) is
981, it is at the numbered CGMCC that registers on the books at China Committee for Culture Collection of Microorganisms's common micro-organisms center
No.10802；Described fermentation medium B is made up of water and solute, and described solute is made up of bean sprout juice and NaCl, described bean sprout juice
It is from bean sprout, to extract, with water, the water-soluble substances obtained；In described fermentation medium B, the content of NaCl is 0.02-0.1g/L,
The content of bean sprout juice is 50-150g/L in terms of bean sprout.
5. the photosynthetic bacteria microbial inoculum prepared by preparation method described in claim 4.
6. contain the animal feed of photosynthetic bacteria microbial inoculum described in claim 1 or 5.
7. described in photosynthetic bacteria microbial inoculum described in claim 1 or 5, the preparation method of claim 2,3 or 4 or claim 6
Animal feed preparation improve chicken immune performance and/or improve chicken production performance and/or improve chicken meat quality product in
Application.
8. described in photosynthetic bacteria microbial inoculum described in claim 1 or 5, the preparation method of claim 2,3 or 4 or claim 6
Animal feed improving chicken production performance and/or improving chicken meat quality and/or improve the application in chicken immune performance.
9. according to the application described in claim 7 or 8, it is characterised in that: described raising chicken immune performance is following I1)-I5) in
All or part of:
I1) broiler thymus index is improved；
I2) broiler index and spleen index is improved；
I3) broiler bursal index is improved；
I4) broiler erythrocyte C is improved3bReceptor rosette rate；
I5) broiler red blood cell immune complexes rosett is improved；
Described production performance is following P1)-P6) in all or part of:
P1) rate of animals delivered to the slaughter-house；
P2) meat feed ratio；
P3) apparent digestibility of protein；
P4) apparent digestibility of calcium；
P5) apparent digestibility of phosphorus；
P6) body weight；
Described raising chicken meat quality is following Q1)-Q8) in all or part of:
Q1) valine content of Carnis Gallus domesticus is improved；
Q2) methionine content of Carnis Gallus domesticus is improved；
Q3) phenylalanine content of Carnis Gallus domesticus is improved；
Q4) cystine of Carnis Gallus domesticus is improved；
Q5) tyrosine content of Carnis Gallus domesticus is improved；
Q6) crude protein content of Carnis Gallus domesticus is improved；
Q7) cholesterol level of Carnis Gallus domesticus is reduced；
Q8) fat content of Carnis Gallus domesticus is reduced.
10. improve chicken production performance and/or improve chicken meat quality and/or the method improving chicken immune performance, including with containing photosynthetic
Antibacterial microbial inoculum Diet chicken；Described containing photosynthetic bacteria microbial inoculum daily ration photosynthetic by described in the common daily ration of chicken and claim 1 or 5
Antibacterial microbial inoculum forms；The common daily ration of described chicken does not contains described photosynthetic bacteria microbial inoculum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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